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23260235

Epigallocatechin gallate attenuates interstitial cystitis in human bladder urothelium cells by modulating purinergic receptors.

Epigallocatechin gallate (EGCG) has exhibited antitumor properties against bladder cancer. However, its effects in interstitial cystitis (IC) have not been investigated. Here, we performed repeated cystoscopy and re-biopsy of bladder mucosa before and after intravesical irrigation of EGCG in eight patients diagnosed with IC based on clinical and histopathologic assessments. Six normal bladder tissue samples were obtained from age-, race-, and sex-matched asymptomatic control subjects. IC symptom index was used to compare the therapeutic effect in IC patients. Patient-derived bladder epithelial cells were cultured and cell stretch experiments and ATP assays were performed. The expression of purinergic receptors X1, X2, and X3, and Y1, Y2, and Y11, in biopsied samples was detected by Western blotting and real-time polymerase chain reaction, respectively. Moreover, the expression of inducible NO synthase, phosphorylated Akt, and phosphorylated NF-κB was also assessed. All EGCG-treated patients demonstrated different extents of remission of symptoms. We found a significant upregulation in P2X1, P2X2, and P2X3 receptor proteins and P2Y1, P2Y2, and P2Y11 receptor transcripts in IC patients. However, EGCG therapy attenuated the expression of all purinergic receptors. In addition, EGCG demonstrated prominent antioxidative and antiinflammatory effects via inhibition of the upregulation of iNOS and phosphorylated NF-κB. Furthermore, the stretch-activated release of ATP in cultured bladder urothelial cells was greater in cells derived from IC patients, compared with those from the control patients, but EGCG, at all concentrations tested, effectively abolished the increase in ATP release from stretched IC patient-derived cells. Our study suggests that inhibition of the expression of purinergic receptors and ATP release in urothelial cells by EGCG supports further development of EGCG as a novel therapeutic option for IC.

23260234

IL-18 activation is dependent on Toll-like receptor 4 during renal obstruction.

Interleukin 18 (IL-18) is a critical mediator of obstruction-induced renal injury. Although previous studies have demonstrated that IL-18 participates in a positive feedback loop via the IL-18 receptor (IL-18R) and localized renal IL-18 and IL-18R production to tubular epithelial cells (TEC), the mechanism of IL-18 activation during obstruction remains unclear. We hypothesized that IL-18 activation is dependent on Toll-like receptor 4 (TLR4) signaling during renal obstruction. Male C57BL6 TLR4 knockout (TLR4KO) and wild-type (WT) mice were subjected to unilateral ureteral obstruction versus sham operation for 1 wk. The animals were sacrificed, and renal cortical tissue was harvested and analyzed for TLR4 expression (Western blot), active IL-18 production (enzyme-linked immunosorbent assay, real-time polymerase chain reaction), IL-18 receptor expression (real-time polymerase chain reaction), and TLR4/IL-18 versus IL-18R cellular localization (dual immunofluorescent staining). Renal TLR4 expression increased significantly in WT mice in response to obstruction, but remained at sham treatment levels in TLR4KO mice. IL-18 and IL-18R gene expression and active IL-18 production were similarly increased in WT mice in response to obstruction, but decreased significantly to sham treatment levels in the absence of TLR4. Dual immunofluorescent staining revealed co-localization of TLR4 and IL-18 to renal TEC during obstruction. IL-18 production and activation during renal obstruction is dependent on intact TLR4 signaling. Co-localization of IL-18 and TLR4 production to TEC during obstruction suggests that TEC are the primary site of IL-18 production and activation. Further characterization of the pathway may be necessary to develop targeted therapy in obstruction-induced renal injury.

23260233

Temporal trends in outcomes following sublobar and lobar resections for small (≤ 2 cm) non-small cell lung cancers--a Surveillance Epidemiology End Results database analysis.

Since the randomized, controlled study that favored lobectomy for resection of stage I non-small cell lung cancers (NSCLCs) by the Lung Cancer Study Group, there have been improvements in staging. The liberal use of computed tomography also may have altered the types of early lung cancer diagnosed. Studies published since then have drawn contradictory conclusions on the benefit of lobectomy over sublobar resections for early-stage NSCLC. We examined the Surveillance Epidemiology End Results database to test our hypothesis that the relationship between extent of resection and outcome has changed since the Lung Cancer Study Group study was published. We examined stage I NSCLCs ≤ 2 cm in size over three periods: 1988-1998 (Early), 1999-2004 (Intermediate), and 2005-2008 (Late). For each period, we assessed overall and disease-specific survivals and their associations with the extents of resection, by univariate and multivariate analyses. Sublobar resections in the Early group could not be categorized into segmentectomies and wedge resections because these were not coded separately. The proportion of NSCLCs ≤ 2 cm increased from 0.98% in 1988 to 2.2% in 2008. Multivariate analyses showed that sublobar resection was inferior to lobectomy in the Early period (hazard ratio [HR], 1.41; 95% confidence interval [CI], 1.21-1.65). This effect decreased in the Intermediate period, in which segmentectomies but not wedge resections were equivalent to lobectomies (wedge versus lobectomy HR, 1.19; 95% CI, 1.01-1.41; segmentectomy versus lobectomy HR, 1.04; 95% CI, 0.8-1.36). The difference disappeared in the Late period, when both wedge resections and segmentectomies were equivalent to lobectomy (wedge versus lobectomy HR, 1.09; 95% CI, 0.79-1.5; segmentectomy versus lobectomy HR, 0.83; 95% CI, 0.47-1.45). Trends for both overall survival and disease-specific survival were identical. The survival benefit of lobectomy over sublobar resection decreased over the past 2 decades with no discernible difference in the most contemporary cases. These results support reevaluation of lobectomy as the standard of care for small (≤ 2-cm) NSCLCs.

23260232

Human platelets promote liver regeneration with Kupffer cells in SCID mice.

Platelets contain several growth factors, including vascular endothelial growth factor (VEGF) and insulin-like growth factor. We examined the role of human platelets in liver regeneration with a focus on Kupffer cells (KCs). Severe combined immunodeficiency mice were subjected to 70% hepatectomy and phosphate-buffered saline administration (PBS); 70% hepatectomy and human platelet transfusion (hPLT); 70% hepatectomy, KC depletion, and PBS administration (KD + PBS); 70% hepatectomy, KC depletion, and human platelet transfusion (KD + hPLT); or a sham operation and human platelet transfusion (sham). The groups were evaluated for liver regeneration, accumulation and activation of human platelets in the liver, and/or co-localization of platelets and KCs. The liver-to-body weight ratio was significantly higher 48 h post-transfusion in the hPLT group compared with the PBS, KD + PBS, and KD + hPLT groups. Human VEGF concentrations were higher in liver tissues from the hPLT group, whereas VEGF was not detected in the other groups. Hepatic levels of KC-derived cytokines were elevated in the hPLT group compared with the PBS group. Molecules in signaling cascades downstream of these cytokines were phosphorylated earlier and more robustly in the hPLT group than in the PBS group. Activated human platelets accumulated in livers in the hPLT group, whereas fewer platelets accumulated and many were not activated in the sham and KD + hPLT groups. In the hPLT group, most human platelets were attached to KCs. Human platelet transfusion promoted liver regeneration in severe combined immunodeficiency mice. Together, human platelets and KCs resulted in growth factor release and enhanced liver regeneration.

23260231

Biphasic scaffolds for repair of deep osteochondral defects in a sheep model.

To oppose the disadvantages of autologous osteochondral transplantation in the treatment of deep osteochondral defects such as donor site morbidity, size limitation, and insufficient chondral integration, we developed two biphasic scaffolds of either hydroxylapatite/collagen (scaffold A) or allogenous sterilized bone/collagen (scaffold B) and tested their integration in a sheep model. We collected chondral biopsies from 12 sheep for the isolation of chondroblasts and cultured them for 4 wk. We created defects at the femoral condyle and implanted either scaffold A or B with chondrocytes or cell free. After 6 wk, animals were euthanized, we explanted the condyles, and evaluated them using histological, immunohistochemical, molecular biological, and histomorphometrical methods. Specimens with scaffold A showed severe lowering of the surface, and the defect size was larger than for scaffold B. We found more immune-competent cells around scaffold A. Chondrocytes were scarcely detected on the surface of both scaffolds. Histomorphometry of the interface between scaffold and recipient showed no significant difference regarding tissue of chondral, osseous, fibrous or implant origin or tartrate-resistant acid phosphatase-positive cells. Real-time reverse transcriptase-polymerase chain reaction analysis revealed significant up-regulation for collagen II and SOX-9 messenger ribonucleic acid expression on the surface of scaffold B compared with scaffold A. Scaffold B proved to be stable and sufficiently integrated in the short term compared with scaffold A. More extensive evaluations with scaffold B appear to be expedient.

23260229

Selective serotonin reuptake inhibitors inhibit human osteoclast and osteoblast formation and function.

Selective serotonin reuptake inhibitors (SSRIs) are widely used antidepressants and one of the most commonly used medications. There is growing concern that SSRIs, which sequester in bone marrow at higher concentrations than brain or blood, increase bone fragility and fracture risk. However, their mechanism of action on human osteoclasts (OC) and osteoblasts (OB) differentiation remains unclear. Expression of serotonin receptors (5-HTR), transporter (5-HTT), and tryptophan hydroxylase 1 (TPH1) was assessed in human OC (precursors and mature) and OB (nonmineralizing and mineralizing) by polymerase chain reaction. OC formation and resorption was measured in the presence of 5 SSRIs. OBs cultured with SSRIs for 28 days were assessed for alkaline phosphatase (ALP) and bone mineralization. Cell viability and apoptosis were determined by annexin V flow cytometry. OCs and OB expressed TPH1, 5-HTT, and 5-HTR1B. The 5-HTR2A was expressed only in OB, whereas 5-HTR2B expression increased from precursor to mature OC. All SSRIs (except citalopram) dose-dependently inhibited OC formation and resorption between 1 μmol/L and 10 μmol/L; order of potency: sertraline > fluoxetine > paroxetine > fluvoxamine > citalopram. Similarly, SSRIs (except citalopram) inhibited ALP and bone mineralization by OB but only at 30 μmol/L. Apoptosis was induced by SSRIs in OC and OB in an identical pattern to inhibitory effects. Serotonin treatment had no effect on either OC or OB parameters. These data demonstrate that SSRIs differentially inhibit bone cell function via apoptosis. This may explain the mechanisms of bone loss with chronic use and aid clinical choices.

23260228

The stability of NR2B in the nucleus accumbens controls behavioral and synaptic adaptations to chronic stress.

The nucleus accumbens (NAc) is closely correlated with depression. It has been demonstrated that the glutamatergic system in NAc plays an important role in the reward pathway, dysfunction of which would cause anhedonia, a core symptom of depression. We therefore tested whether N-methyl-D-aspartate receptors and the synaptic plasticity in the NAc are regulated by chronic stress and the relevance to depression. We applied behavioral tests (n = 12, each group) of social interaction and sucrose preference tests to identify the susceptibility of mice to chronic social defeat stress. We then tested N-methyl-D-aspartate receptor-long-term depression at cortico-accumbal synapse to determine the relationship between the susceptibility and changes in synaptic plasticity (n = 8, each group). We further investigated whether restoration of these changes could produce antidepressant effects (n = 10). We found that chronic stress induced selective downregulation of N-methyl-D-aspartate receptor NR2B subunits in the confined surface membrane pool of NAc neurons. Remarkably, the loss of synaptic NR2B was a long-lived event and further translated to the significant modulation of synaptic plasticity in the form of long-term depression. We further observed that the stress-induced changes were restored by fluoxetine and that resilient mice-those resistant to chronic stress-showed patterns of molecular regulation in the NAc that overlapped dramatically with those seen with fluoxetine treatment. Behaviorally, restoration of NR2B loss prevented the behavioral sensitization of mice to chronic stress. Our results identify NR2B in the NAc as a key regulator in the modulation of persistent psychomotor plasticity in response to chronic stress.

23260227

Sexually dimorphic placental development throughout gestation in the spiny mouse (Acomys cahirinus).

It has been hypothesized that male fetuses down regulate placental growth during periods of accelerated fetal growth. We aimed to investigate this, and determine whether sexual dimorphism was apparent in the spiny mouse placenta. We hypothesized that expression of fetal growth promoters would be higher in placentas of males, whereas genes involved in placental structural development would be more highly expressed in placentas of females. Spiny mouse dams, a precocial rodent with an in utero endocrine milieu dissimilar from other rodents, but akin to humans, were sacrificed at gestational ages 15-37 (term = 39 days). Placentas were collected and processed for histology or qPCR analysis of selected genes (GCM1, MAP2K1, SLC2A1, NR3C1, IGF1, IGF1R). Fetal and placental weights were similar for both sexes. Placentas of female fetuses had less spongy zone (P(SEX) < 0.0001), and more labyrinth (P(SEX) < 0.0001) than males. Early placenta and labyrinth expression of SLC2A1 was higher in males than females (P(SEX) < 0.05). Labyrinthine IGF1R remained constant until term in the female, compared with male where expression increased until term. Peak MAP2K1 expression occurred earlier in the male placenta than the female. Spongy zone SLC2A1 remained constant until term in the female, compared with male where expression increased until term. The spiny mouse is a species that exhibits sexually dimorphic placental development. We suggest that these sex differences in placental gene expression and structure may underlie or compound the male vulnerability to a sub-optimal in utero environment.

23260200

Regulation of myofibroblast differentiation by poly(ADP-ribose) polymerase 1.

Poly(ADP-ribosyl)ation (PARylation) is a post-translational protein modification effected by enzymes belonging to the poly(ADP-ribose) polymerase (PARP) superfamily, mainly by PARP-1. The key acceptors of poly(ADP-ribose) include PARP-1 itself, histones, DNA repair proteins, and transcription factors. Because many of these factors are involved in the regulation of myofibroblast differentiation, we examined the role of PARylation on myofibroblast differentiation. Overexpression of PARP-1 with an expression plasmid activated expression of the α-SMA gene (Acta2), a marker of myofibroblast differentiation in lung fibroblasts. Suppression of PARP-1 activity or gene expression with PARP-1 inhibitors or siRNA, respectively, had the opposite effect on these cells. PARP-1-deficient cells also had reduced α-SMA gene expression. DNA pyrosequencing identified hypermethylated regions of the α-SMA gene in PARP-1-deficient cells, relative to wild-type cells. Interestingly, and of potential relevance to human idiopathic pulmonary fibrosis, PARP activity in lung fibroblasts isolated from idiopathic pulmonary fibrosis patients was significantly higher than that in cells isolated from control subjects. Furthermore, PARP-1-deficient mice exhibited reduced pulmonary fibrosis in response to bleomycin-induced lung injury, relative to wild-type controls. These results suggest that PARylation is important for myofibroblast differentiation and the pathogenesis of pulmonary fibrosis.

23260201

Adenovirus serotype 14 infection, New Brunswick, Canada, 2011.

We describe 3 culture-proven cases of adenovirus serotype 14 infection in New Brunswick, Canada, during the summer of 2011. Strains isolated from severely ill patients were closely related to strains of a genomic variant, adenovirus 14p1, circulating in the United States and Ireland. Physicians in Canada should be aware of this emerging adenovirus.

23260199

Human sarcomas are mosaic for telomerase-dependent and telomerase-independent telomere maintenance mechanisms: implications for telomere-based therapies.

Telomere shortening necessitates that tumor cells activate a telomere maintenance mechanism (TMM) to support immortalization. Although most tumor cells activate expression of the enzyme telomerase, some cells elongate telomeres in a telomerase-independent manner, termed alternative lengthening of telomeres (ALT). Previous studies have evaluated the presence of telomerase or ALT mechanisms or both in a variety of tumor types. Our studies also show that TMMs are not mutually exclusive in some tumors. In contrast, our IHC analyses of human sarcomas identified a subset of tumors with some cells containing ALT-associated PML bodies, a hallmark of ALT, and separate cells expressing telomerase in the same tumor. By using a second set of human osteosarcomas, we merged IHC and biochemical analyses to characterize more fully the tumor TMM. The IHC data reveal the presence of both telomerase- and ALT-positive tumor cells in samples that demonstrate characteristics of both telomerase and ALT in biochemical assays. These assays, which measure telomere length and telomerase activity of tumor extracts, are conventionally used to classify tumor TMM. Our results suggest that TMM is not a single or perhaps static characteristic of some tumors and that TMM heterogeneity should be considered in tumor stratification. Furthermore, clinical interest in telomere-based therapies may necessitate accurate characterization of tumor TMM before treatment to maximize therapeutic efficacy.

23260197

Is there a maximal anabolic response to protein intake with a meal?

Several recent publications indicate that the maximum stimulation of muscle protein fractional synthetic rate occurs with intake of 20-30 g protein. This finding has led to the concept that there is a maximal anabolic response to protein intake with a meal, and that the normal amount of protein eaten with dinner will generally exceed the maximally-effective intake of protein. However, protein breakdown has not been taken into account when evaluating the anabolic response to protein intake. Protein anabolism occurs only when protein synthesis exceeds protein breakdown. Higher protein intakes when protein synthesis is maximized is characterized by suppressed protein breakdown and via that mechanism leads to a greater anabolic response. This explains why when net protein synthesis is measured, the relationship between amino acid availability and net gain remains linear, without any apparent plateau of effect at higher levels of availability. We conclude that there is no practical upper limit to the anabolic response to protein or amino acid intake in the context of a meal.

23260195

Distribution and compartmentalization of human circulating and tissue-resident memory T cell subsets.

Knowledge of human T cells derives chiefly from studies of peripheral blood, whereas their distribution and function in tissues remains largely unknown. Here, we present a unique analysis of human T cells in lymphoid and mucosal tissues obtained from individual organ donors, revealing tissue-intrinsic compartmentalization of naive, effector, and memory subsets conserved between diverse individuals. Effector memory CD4(+) T cells producing IL-2 predominated in mucosal tissues and accumulated as central memory subsets in lymphoid tissue, whereas CD8(+) T cells were maintained as naive subsets in lymphoid tissues and IFN-γ-producing effector memory CD8(+) T cells in mucosal sites. The T cell activation marker CD69 was constitutively expressed by memory T cells in all tissues, distinguishing them from circulating subsets, with mucosal memory T cells exhibiting additional distinct phenotypic and functional properties. Our results provide an assessment of human T cell compartmentalization as a new baseline for understanding human adaptive immunity.

23260196

Caspase-8 blocks kinase RIPK3-mediated activation of the NLRP3 inflammasome.

Caspase-8 deficiency in certain cells prompts chronic inflammation. One mechanism suggested to account for this inflammation is enhanced signaling for necrotic cell death, mediated by the protein kinases RIPK1 and RIPK3 that caspase-8 can cleave. We describe an activity of caspase-8 in dendritic cells that controls the initiation of inflammation in another way. Caspase-8 deficiency in these cells facilitated lipopolysaccharide-induced assembly and function of the NLRP3 inflammasome. This effect depended on the functions of RIPK1 and RIPK3, as well as of MLKL and PGAM5, two signaling proteins recently shown to contribute to RIPK3-mediated induction of necrosis. However, although enhancement of inflammasome assembly in the caspase-8-deficient cells shares proximal signaling events with the induction of necrosis, it occurred independently of cell death. These findings provide new insight into potentially pathological inflammatory processes to which RIPK1- and RIPK3-mediated signaling contributes.

23260194

Pathogen-specific inflammatory milieux tune the antigen sensitivity of CD8(+) T cells by enhancing T cell receptor signaling.

CD8(+) T cells confer host protection through T-cell-receptor (TCR)-mediated recognition of foreign antigens presented by infected cells. Thus, generation of CD8(+) T cell populations with high antigen sensitivity is critical for efficient pathogen clearance. Besides selection of high-affinity TCRs, the molecular mechanisms regulating the antigen sensitivity of CD8(+) T cells remain poorly defined. Herein, we have demonstrated that the antigen sensitivity of effector and memory CD8(+) T cells is dynamically regulated and can be tuned by pathogen-induced inflammatory milieux independently of the selection of cells with higher TCR affinity. Mechanistically, we have demonstrated that the signal-transduction capacity of key TCR proximal molecules is enhanced by inflammatory cytokines, which reduced the antigen density required to trigger antimicrobial functions. Dynamic tuning of CD8(+) T cell antigen sensitivity by inflammatory cytokines most likely optimizes immunity to specific pathogens while minimizing the risk of immunopathology at steady state.

23260193

Cytokine-mediated programmed proliferation of virus-specific CD8(+) memory T cells.

During infection, CD8(+) T cells not only respond to antigenic signals through their T cell receptor (TCR) but also incorporate inflammatory signals from cytokines produced in the local infected microenvironment. Transient TCR-mediated stimulation will result in programmed proliferation that continues despite removal of the antigenic stimulus, but it remains unclear whether brief exposure to specific cytokines will elicit similar effects. Here, we have demonstrated that brief stimulation of memory T cells with interleukin-12 (IL-12) and interleukin-18 (IL-18) results in tightly regulated programmed proliferation, in addition to acquisition of enhanced virus-specific cytokine production and cytolytic activity. CD8(+) T cells briefly exposed to IL-12 and IL-18 in vitro showed improved antiviral activity in vivo, as demonstrated by increased proliferation and reduced viremia. These results indicate that even transitory exposure to inflammatory cytokines can provide a selective advantage to infiltrating CD8(+) T cells by triggering a developmental program that is initiated prior to direct contact with virus-infected cells.

23260188

A multilevel intervention to increase community hospital use of alteplase for acute stroke (INSTINCT): a cluster-randomised controlled trial.

Use of alteplase improves outcome in some patients with stroke. Several types of barrier frequently prevent its use. We assessed whether a standardised, barrier-assessment, multicomponent intervention could increase alteplase use in community hospitals in Michigan, USA. In a cluster-randomised controlled trial, we selected adult, non-specialty, acute-care community hospitals in the Lower Peninsula of Michigan, USA. Eligible hospitals discharged at least 100 patients who had had a stroke per year, had less than 100 000 visits to the emergency department per year, and were not academic comprehensive stroke centres. Using a computer-generated randomisation sequence, we selected 12 matched pairs of eligible hospitals. Within pairs, the hospitals were allocated to intervention or control groups with restricted randomisation in January, 2007. Between January, 2007, and December, 2007, intervention hospitals implemented a multicomponent intervention that included qualitative and quantitative assessment of barriers to alteplase use and ways to address the findings, and provided additional support. The primary outcome was change in alteplase use in patients with stroke in emergency departments between the pre-intervention period (January, 2005, to December, 2006) and the post-intervention period (January, 2008, to January, 2010). Physicians in participating hospitals and the coordinating centre could not be masked to group assignment, but were masked to progress made in paired control hospitals. External medical reviewers who were masked to group assignment assessed outcomes. We did intention-to-treat (ITT) and target-population (without one pair that was excluded after randomisation) analyses. This trial is registered at ClinicalTrials.gov, number NCT00349479. All 24 hospitals completed the study. Overall, 745 of 40 823 patients with stroke received intravenous alteplase treatment. In the ITT analysis, the proportion of patients with stroke who were admitted and treated with alteplase increased between the pre-intervention and post-intervention periods in intervention hospitals (89 [1·25%] of 7119 patients to 235 [2·79%] of 8419) to a greater extent than in control hospitals (99 [1·25%] of 7946 to 194 [2·10%] of 9222), but the difference between groups was not significant (relative risk [RR] 1·37, 95% CI 0·96-1·93; p=0·08). In the target-population analysis, the increase in alteplase use in intervention hospitals (59 [1·00%] of 5882 to 191 [2·62%] of 7288) was significantly greater than in control hospitals (65 [1·09%] of 5957 to 120 [1·72%] of 6989; RR 1·68, 95% CI 1·09-2·57; p=0·02), but was still clinically modest. The intervention did not significantly increase alteplase use in patients with ischaemic stroke. The increase in use of alteplase in the target population was significant, but smaller than the effect to which the study was powered. Additional strategies to increase acute stroke treatment are needed. National Institutes of Health National Institute of Neurological Disorders and Stroke.

23260189

Clinical features, diagnosis, and treatment of human African trypanosomiasis (sleeping sickness).

Human African trypanosomiasis, or sleeping sickness, is caused by infection with parasites of the genus Trypanosoma, transmitted by the tsetse fly. The disease has two forms, Trypanosoma brucei (T b) rhodesiense and T b gambiense; and is almost always fatal if untreated. Despite a recent reduction in the number of reported cases, patients with African trypanosomiasis continue to present major challenges to clinicians. Because treatment for CNS-stage disease can be very toxic, diagnostic staging to distinguish early-stage from late-stage disease when the CNS in invaded is crucial but remains problematic. Melarsoprol is the only available treatment for late-stage T b rhodesiense infection, but can be lethal to 5% of patients owing to post-treatment reactive encephalopathy. Eflornithine combined with nifurtimox is the first-line treatment for late-stage T b gambiense. New drugs are in the pipeline for treatment of CNS human African trypanosomiasis, giving rise to cautious optimism.

23260186

[Association between the myeloperoxidase gene polymorphisms and the susceptibility to prostate cancer: a case-control study in a Chinese population].

Prostate cancer (PCa) is the most common cancer among men in most western populations. The polymorphisms of the myeloperoxidase (MPO) gene have been correlated with abnormal MPO expression and increased risk of various types of cancers. Our study aimed to evaluate the association between the genetic polymorphisms and the risk of prostate cancer. Genotyping was carried out by using the genotyping system (MassARRAY iPLEX; Sequenom, Inc., San Diego, CA, USA) on 1,108 PCa patients and 1,525 cancer-free controls in a Chinese Han population. Although one SNP (rs8082134, P < 0.050) was significant, it is very rare and unstable. Other SNPs had no significant difference between genotype distributions in the PCa patients and the control group. Totally, SNPs in the MPO gene is not associated with PCa risk. Our data showed a limited association between the MPO SNPs and the susceptibility to PCa in population of Chinese Han population. The possible association of rs8082134 of MPO with PCa risk need further clarification.

23260185

Estimation of bladder cancer projections in Spain.

One of the most frequent cancers in the world is bladder cancer that affects, according to some authors, 5.4 million persons in the most developed countries. Our study has aimed to estimate the impact projections of this disease in Spain between 2007 and 2022. Mortality data for bladder cancer mortality was used International Classification of Diseases 10th revision (ICD-10 C67), and the population data from 1998 to 2007. The data were obtained from the National Institute of Statistics (INE). Relative survival was obtained form the EUROCARE study. The projections of incidence, prevalence, and mortality were estimated using the statistical program Mortality-Incidence Analysis MODel (MIAMOD) and after the joinpoint regression that calculates the Annual Percentage of Change (APC). Between the years 1998 and 2022, it is foreseen that there will be a decrease in prevalence in the men, from 156.93 (adjusted rate AR=128.71) to 132.99 (AR=84.68) cases per 100,000 inhabitants/year in the year 2022. The incidence rate would decrease from 30.2 (AR=24.93) to 24.87 (AR=15.88) and mortality from 14.96 (AR=12.25) to 12.08 (AR=7.67). In women, an increase is expected in prevalence from 21.18 (AR=13.23) to 35.6 (AR=21.46) cases per 100.000 inhabitants/year. The incidence of 4.8 (AR=2.91) in 1998 to 7.79 (AR=4.69), Mortality will increase from 2.25 (AR=1.32) to 3.37 (AR=1.89) between the years 1998 and 2022. In men, the APCs found for prevalence, incidence and mortality were, respectively, 0.69 (95% CI%:-6.9/8.9); 0.69 (95% CI:-6.9/8.95) and -4.18 (95% CI:-11.32/3.51), these not being statistically significant. In women, the APC for prevalence was -4.44 (95% CI:-13.4/5.44). from 19.24 (95% CI:8.93/30.52).and from -3.28 (95% CI:-20.26/17.20) for mortality. This cancer should be monitored and in turn prevented based on the controllable risk factors, especially in women.

23260184

[Available evidence about efficacy of different restoring agents of glycosaminoglycans for intravesical use in interstitial cystitis].

To compare the different endovesical therapeutic regimes in terms of clinical effectiveness based on glycosaminoglycan replenishment agents (RA-GAG) available on the market in Spain. A bibliographic analysis was made of the studies published in Medline from 1996 to 2012 on RA-GAG of application in the bladder, placing emphasis on the clinical results. A post-hoc comparison was made of the efficacy of this treatment in the studies conducted in patients with interstitial cystitis in different conditions by calculating the effect sizes to analyze improvement on the pain visual analogue scale (VAS) and clinical response rate. The number of patients needed to treat (NNT) for the different agents was calculated based on the odds ratio and associated economic implications. The globally available evidence is scarce. There are 38 articles about RA-GAGs in different indications, 71 of them in interstitial cystitis and only 8 may assist in establishing a comparison between the results presented. The treatments used were placebo, 0.8% high molecular weight hyaluronic acid (Cystistat(®)), 2% chondroitin sulfate sodium (Uracyst(®)) and a combination of 1.6% low molecular weight hyaluronic acid plus 2% chondroitin sulfate (Ialuril(®)), between 6 and 12 instillations. Another low molecular weight hyaluronic acid preparation (Uromac(®)) lacks any scientific evidence. All the therapeutic elements studied show a mean score decrease on the pain VAS and increase in the rate of post-treatment response. The NNT for the treatments that are statistically more beneficial over placebo ranges from 1.6 and 4.1. The post-hoc comparison of the response rates has established that Cystistat(®) 12 instillations (OR 18.8; 95% CI 6.4-57.2; P=.001) or 10 instillations (OR 19.2; 95% CI 5.3-75.3; P=.001) are the treatment regimes that obtain maximum effectiveness. In both cases, the NNT was 1.6. This study has multiple limitations inherent to the nature of the design. However, although the available literature is scarce, it shows that there are differences regarding the clinical effectiveness of the different agents and regimes used for endovesical treatment of interstitial cystitis. These differences also entail economic type implications.

23260183

[Prognostic factors in emphysematous pyelonephritis].

The purpose of this study is to analyze our experience with 18 cases of Emphysematous pyelonephritis (EPN) in a tertiary care center and describe our treatment strategy. Of 262 patients admitted with acute pyelonephritis, 18 had CT findings of EPN. The Wan and Huang classifications were used. We assessed the clinical, radiological, and therapeutic characteristics of these patients and investigated potential prognostic factors of mortality. Between 2005 and 2010, 17 women and 1 man with EPN were treated. Mean age was 52.4 years. Diabetes was found in 66% and hypertension in 72%. The most common clinical findings were tachycardia (11), fever (11) and flank pain (9); 66% (12) presented with severe sepsis and 2 had septic shock. Acute renal injury developed in 61%. Nine patients were treated exclusively with conservative management; 5 had double J stenting, 3 had CT-guided PCD and 1 required nephrectomy after unsuccessful medical management. Mortality was 11%. Altered consciousness (P=.0001), multiple organ failure (P=.0004), hyperglycemia (P=.003) and elevated leukocyte count (> 20000 K) (P=.01) were more frequent among patients dying from EPN. No difference in mortality was found between patients managed conservatively and those undergoing invasive therapy. Although rare, EPN should be suspected in patients with multiple comorbidities presenting with severe sepsis. Altered consciousness, multiple organ failure, hyperglycemia and elevated leukocyte count are poor prognosis indicators. Invasive management should be used judiciously and medical treatment can be a safe strategy in selected cases.

23260182

Biochemical and behavioural responses of the endobenthic bivalve Scrobicularia plana to silver nanoparticles in seawater and microalgal food.

Because of their bactericidal effects, Ag nanoparticles (Ag NPs) have promising industrial development but could lead to potential ecological risks. The aim of this study was to examine the uptake and effect of silver (soluble or as lactate Ag NPs of 40 nm) at low concentrations (10 μg L(-1)) in the endobenthic bivalve Scrobicularia plana exposed, for 14 days, directly (water) or via the diet (microalgae). The stability of Ag NPs in seawater was examined using dynamic light scattering. Release of soluble Ag from Ag NPs in the experimental media was quantified by using diffusive gradient in thin film. Bioaccumulation of Ag in bivalves was measured by electrothermal atomic absorption spectrometry. Behavioural and biochemical biomarkers were determined in bivalves. Aggregation of Ag NPs and the release of soluble Ag from Ag NPs were observed in the experimental media. For both forms of Ag, bioaccumulation was much more important for waterborne than for dietary exposure. The response of oxidative stress biomarkers (catalase, glutathion S-transferase, superoxide dismutase) was more important after dietary than waterborne exposure to Ag (soluble and NPs). These defences were relatively efficient since they led to a lack of response of damage biomarkers. Burrowing was not affected for bivalves exposed directly or through the diet to both Ag forms but feeding behaviour was impaired after 10 days of dietary exposure. Since no differences of responses to Ag either soluble or nanoparticulate were observed, it seems that labile Ag released from Ag NPs was mainly responsible for toxicity.

23260181

Lethal and sublethal effects of cadmium in the white shrimp Palaemonetes argentinus: a comparison between populations from contaminated and reference sites.

In the present study, the acute toxicity of cadmium (Cd) in white shrimp (Palaemonetes argentinus) from a metal polluted lagoon (Los Padres, LP) and from unpolluted lagoon (Nahuel Ruca, NR) was evaluated. Both population, were exposed to 3.06, 12.26, 30.66, 61.32, 306 and 613.2 μg Cd · L(-1) for 96-h. The sublethal effects of Cd were examined by two cellular biomarkers: metallothionein (MT) and lipid peroxidation (LPO). The seasonal variations of biomarkers in both lagoons were also evaluated. P. argentinus demonstrated a high sensitivity to Cd, with values of 96-h LC50 lower and close to those of highly sensitive species; therefore, can be proposed as a good indicator species. The LC(50) values of shrimp from LP (24-h: 269.8, 48-h: 67.45, 72-h: 30.66, 96-h: 24.50 μg Cd · L(-1)) were higher than those from NR (24-h: 153.3, 48-h: 32.65, 72-h: 18.40, 96-h: 12.26 μg Cd · L(-1)), indicating a higher tolerance to Cd, and it was related to their origin. Differential responses in terms of MT induction and LPO between populations were also detected. In NR shrimps, the MT synthesis was induced very fast (24-h) and even at the minimum concentration tested (3.06 μg Cd · L(-1)), while no increases were observed in LPO levels. In contrast, the MT and LPO levels in LP shrimps were not increased relative to control, although they were more tolerant to Cd than those of NR; suggesting the presence of another mechanism involved in the detoxification of Cd. The differences in both sensitivity and biochemical responses to Cd may be related with their environmental histories.

23260180

Zinc-induced oxidative damage, antioxidant enzyme response and proline metabolism in roots and leaves of wheat plants.

The purpose of this study was to evaluate antioxidative responses and proline metabolism in roots and leaves of wheat seedlings after treatment with different zinc (Zn) concentrations (0, 0.5, 1 and 3mM) for 6 days. A notable reduction in Zn content was observed in 0.5mM Zn-treated leaves, but a significant elevation in response to 1 and 3mM Zn treatment. Significant increases in Zn levels were observed in roots exposed to all applied Zn concentration. The highest Zn concentration resulted in significant reduction in the amount of total chlorophyll (chl) and chl a, while chl b content decreased under all applied Zn concentrations. In wheat leaves, Zn excess caused an insignificant enhancement of hydrogen peroxide (H(2)O(2)) content as well as unaltered malondialdehyde (MDA) level. Unchanged superoxide dismutase (SOD) activity, increased peroxidase (POD), catalase (CAT), glutathione reductase (GR) and ascorbate peroxidase (APX) activities were also observed in the leaves of Zn-treated seedlings. By contrast, higher H(2)O(2) and MDA contents in Zn-treated roots were correlated with the stimulation of SOD and the inhibition of POD and GR. There were significant enhancements of soluble sugar and proline in both leaves and roots of wheat seedlings under Zn stress, but the increased rate of proline was higher in the roots than in the leaves. Differently, soluble protein content due to Zn treatment was lower in the leaves but higher in the roots, as compared with untreated seedlings. Additionally, ornithine δ-aminotransferase in both leaves and roots was stimulated by Zn stress, but different Zn concentrations exhibited inhibitory effect on glutamate kinase activity in wheat seedlings. In contrast, all applied Zn concentration resulted in an elevation of proline dehydrogenase activity in the leaves while the highest Zn concentration inhibited this parameter in the roots.

23260179

Factors that affect the permeability of commercial hollow-fibre membranes in a submerged anaerobic MBR (HF-SAnMBR) system.

A demonstration plant with two commercial HF ultrafiltration membrane modules (PURON(®), Koch Membrane Systems, PUR-PSH31) was operated with urban wastewater. The effect of the main operating variables on membrane performance at sub-critical and supra-critical filtration conditions was tested. The physical operating variables that affected membrane performance most were gas sparging intensity and back-flush (BF) frequency. Indeed, low gas sparging intensities (around 0.23 Nm(3) h(-1) m(-2)) and low BF frequencies (30-s back-flush for every 10 basic filtration-relaxation cycles) were enough to enable membranes to be operated sub-critically even when levels of mixed liquor total solids were high (up to 25 g L(-1)). On the other hand, significant gas sparging intensities and BF frequencies were required in order to maintain long-term operating at supra-critical filtration conditions. After operating for more than two years at sub-critical conditions (transmembrane flux between 9 and 13.3 LMH at gas sparging intensities of around 0.23 Nm(3) h(-1) m(-2) and MLTS levels from around 10-30 g L(-1)) no significant irreversible/irrecoverable fouling problems were detected (membrane permeability remained above 100 LMH bar(-1) and total filtration resistance remained below 10(13) m(-1)), therefore no chemical cleaning was conducted. Membrane performance was similar to the aerobic HF membranes operated in full-scale MBR plants.

23260178

HPLC/HPSEC-FLD with multi-excitation/emission scan for EEM interpretation and dissolved organic matter analysis.

The need to track and characterize dissolved organic matter (DOM) has made fluorescence excitation-emission matrix (EEM) spectroscopy extensively used. In this work, reverse phase high-performance liquid chromatography (RP-HPLC) and high-performance size exclusion chromatography (HPSEC) with fluorescence detector (FLD) were used for EEM interpretation and DOM analysis. Given that fluorescence detectors can scan with multi-excitation or multi-emission mode, HPLC-FLD with multi-excitation scan directly verified the prevalence of multi-peak fluorophores in EEM, which provides a corrective insight for the current fluorescence regional integration (FRI) methods; whereas HPLC-FLD with multi-emission scan provided more informative fluorescence fingerprints for identification of DOM species, which is a chromatographic surrogate for determining the proper number of PARAFAC components. Besides providing a deep insight for the current EEM interpretation, the HPLC/HPSEC-FLD results also directly related physiochemical properties to DOM species, including polarity and molecular weight (MW) distribution, which is helpful for further characterization their behavior in water and wastewater treatment process. A chromatography technique with multi-excitation and multi-emission fluorescence scan can be an informative method for EEM interpretation and DOM identification and characterization.

23260177

Relationship of human-associated microbial source tracking markers with Enterococci in Gulf of Mexico waters.

Human and ecosystem health can be damaged by fecal contamination of recreational waters. Microbial source tracking (MST) can be used to specifically detect domestic sewage containing human waste, thereby informing both risk assessment and remediation strategies. Previously, an inter-laboratory collaboration developed standardized PCR methods for a bacterial, an archaeal, and a viral indicator of human sewage. Here we present results for two subsequent years of field testing in fresh and salt water by five laboratories across the U.S. Gulf Coast (two in Florida and one each in Mississippi, Louisiana and Texas) using common standard operating procedures (SOPs) developed previously. Culturable enterococci were enumerated by membrane filtration, and PCR was used to detect three MST markers targeting domestic sewage: human-associated Bacteroides (HF183), Methanobrevibacter smithii and human polyomaviruses BK and JC (HPyVs). Detection of sewage markers in surface waters was significantly associated with higher enterococci levels and with exceedance of the recreational water quality standard in four or three regions, respectively. Sewage markers were frequently co-detected in single samples, e.g., M. smithii and HF183 were co-detected in 81% of Louisiana samples, and HPyVs and M. smithii were co-detected in over 40% of southwest Florida and Mississippi samples. This study demonstrates the robustness and inter-laboratory transferability of these three markers for the detection of pollution from domestic sewage in the waters impacting the Gulf of Mexico over a coastal range of over 1000 miles.

23260176

Impact of oxygen limitation on glycerol-based biopolymer production by bacterial enrichments.

The increasing production of bioethanol and biodiesel has resulted in the generation of a massive amount of crude glycerol, inducing the need for effective valorization of these waste streams. One of the valorization options could be through conversion of crude glycerol into a biopolymer using microbial community engineering in a feast-famine process. A complicating factor in the production of biopolymers from glycerol encountered in previous works is that two different types of polymers can be formed; polyhydroxyalkanoate (PHA) and polyglucose. Here we describe the effect of limiting the oxygen supply rate on the polymer distribution with the aim of defining the conditions that favour the conversion of glycerol in one single polymer. The decrease of oxygen supply rate during the biopolymer maximization step did not influence glycerol partitioning among PHA and polyglucose, but oxygen limitation during the community enrichment step favoured polyglucose storage over PHA.

23260175

QSAR models for oxidation of organic micropollutants in water based on ozone and hydroxyl radical rate constants and their chemical classification.

Ozonation is an oxidation process for the removal of organic micropollutants (OMPs) from water and the chemical reaction is governed by second-order kinetics. An advanced oxidation process (AOP), wherein the hydroxyl radicals (OH radicals) are generated, is more effective in removing a wider range of OMPs from water than direct ozonation. Second-order rate constants (k(OH) and k(O3) are good indices to estimate the oxidation efficiency, where higher rate constants indicate more rapid oxidation. In this study, quantitative structure activity relationships (QSAR) models for O(3) and AOP processes were developed, and rate constants, k(OH) and [Formula: see text] , were predicted based on target compound properties. The k(O3) and k(OH) values ranged from 5 * 10(-4) to 10(5) M(-1)s(-1) and 0.04 to 18 * (10(9)) M(-1) s(-1), respectively. Several molecular descriptors which potentially influence O(3) and OH radical oxidation were identified and studied. The QSAR-defining descriptors were double bond equivalence (DBE), ionisation potential (IP), electron-affinity (EA) and weakly-polar component of solvent accessible surface area (WPSA), and the chemical and statistical significance of these descriptors was discussed. Multiple linear regression was used to build the QSAR models, resulting in high goodness-of-fit, r(2) (>0.75). The models were validated by internal and external validation along with residual plots.

23260174

Suspended particle and pathogen peak discharge buffering by a surface-flow constructed wetland.

Constructed wetlands (CWs) have been shown to improve the water quality of treated wastewater. The capacity of CWs to reduce nutrients, pathogens and organic matter and restore oxygen regime under normal operating conditions cannot be extrapolated to periods of incidental peak discharges. The buffering capacity of CWs during peak discharges is potentially a key factor for water quality in the receiving waters. Therefore, the aim of the present study was to investigate the behaviour of peak discharges of suspended particles, (associated) physiochemical parameters and pathogenic organisms from a wastewater treatment plant (WWTP) in a full scale constructed wetland (CW). By mixing clarified water and sludge rich water from the settlement tank of the WWTP, the suspended particle concentration was increased for 8 h from ± 3.5 to ± 230 mg L(-1), and discharged into a full scale horizontal surface flow constructed wetland. An increase of suspended particle concentration following the peak discharge concurred with increases in turbidity and oxygen demand, total nutrient load (nitrogen, phosphorus and carbon) and pathogens (Escherichia coli and Enterococci). Temperature, pH, conductivity and dissolved nutrient concentrations (nitrogen, phosphorus and carbon) were however unaffected by the initial peak discharge. After retention in the unvegetated ponds (the first CW compartment) the applied suspended particle peak with a total load of 86.2 kg was reduced by >99%. Similar peak buffering was observed for the turbidity, oxygen demand and settable volume. Simultaneously dissolved nutrient concentrations increased, indicating partial mineralization of the suspended particles during retention in the unvegetated ponds. The peak buffering of pathogens was lower (40-84%), indicating differences in removal processes between other suspended particles and pathogens. The results indicated that the suspended particles were probably mostly removed by sedimentation and mineralization, where pathogens were more likely buffered by biofilm retainment, mortality and predation, mainly in reed ditches. After passing through the total CW the residuals of the suspended particle peak discharge were temporal increased concentrations of inorganic carbon (IC), NH(4) and E. coli (respectively 11%, 17% and 160% higher than steady state concentrations). The observations support the positive role of CWs for effective buffering of wastewater discharge peaks.

23260173

Evaluating the impact of operational parameters on the formation of soluble microbial products (SMP) by activated sludge.

Soluble microbial products (SMP) are the major component of the residual organic fraction in biological wastewater treatment effluent. The impact of process parameters on SMP production by specific groups of bacteria is currently unknown. In this work, SMP production by activated sludge at different substrate concentrations, dissolved oxygen (DO) levels and temperatures, was evaluated by experimental and modeling approaches. The results showed that among the three parameters, SMP production was most sensitive to substrate concentration. Total SMP production was increased 70.5% by a threefold increase in substrate concentration, with SMP produced from heterotrophs, ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) increasing by 61.2%, 580.0% and 410.0%, respectively. The effect of temperature on SMP was less pronounced. Decreasing the temperature from 20 °C to 10 °C decreased total SMP by 17.2%, with SMP production from heterotrophs decreasing by 20.0%, and from the AOB and NOB increasing by 180.0% and 140.0%. DO concentration had nearly no effect on total and heterotrophic SMP production, while it did have a significant positive effect on autotrophic SMP production. SMP production from AOB and NOB decreased by 24.3% and 47.8%, respectively following a decrease in DO concentration from 8.7 to 1.5 mg/L. However, the net effect of DO on total SMP production was negligible.

23260172

Evaluation of high levels of fluoride, arsenic species and other physicochemical parameters in underground water of two sub districts of Tharparkar, Pakistan: a multivariate study.

In present study total arsenic, inorganic arsenic species and fluoride ion contaminations in underground water of Diplo and Chachro sub district of Tharparkar, Pakistan were investigated. The concentrations of total As, inorganic As species, F(-) and others physicochemical parameters were reported in terms of basic statistical parameters, principal component analysis, cluster analysis, sodium absorption ratio and saturation indices. The As(3+) was determined by cloud point extraction using ammonium pyrrolidinedithiocarbamate (APDC) as complexing reagent, and complex was extracted by surfactant-rich phases in the non-ionic surfactant Triton X-114; after centrifugation the surfactant-rich phase was diluted with 0.1 mol/L HNO(3) in methanol. While total inorganic arsenic (iAs) was determined by solid phase extraction using titanium dioxide (TiO(2)) as an adsorbent, after centrifugation, the solid phase was prepared to be slurry for determination. The extracted As species were determined by electrothermal atomic absorption spectrometry. The concentration of As(5+) in the water samples was calculated by the difference of the total iAs and As(3+), while F(-) and other anions were determined by ion chromatography. The positive correlation of F(-) and As species with Na(+) and HCO(3)(-) showed that the water with high salinity and alkalinity stabilized the As species and F(-) in the groundwater. The positive correlation (r = 0.640, p = 0.671) was observed between total As and it species with F(-). Results showed that underground water samples of these two areas of Tharparkar were severely contaminated with arsenic and fluoride ion, which are exceeded the World Health Organization (WHO) provisional guideline value, and United States Environmental Protection Agency, maximum contaminant level of 0.01 mg/L and 1.5 mg/L, respectively.

23260171

A systematic review of studies of depression prevalence in university students.

Depression is a common health problem, ranking third after cardiac and respiratory diseases as a major cause of disability. There is evidence to suggest that university students are at higher risk of depression, despite being a socially advantaged population, but the reported rates have shown wide variability across settings. To explore the prevalence of depression in university students. PubMed, PsycINFO, BioMed Central and Medline were searched to identify studies published between 1990 and 2010 reporting on depression prevalence among university students. Searches used a combination of the terms depression, depressive symptoms, depressive disorders, prevalence, university students, college students, undergraduate students, adolescents and/or young adults. Studies were evaluated with a quality rating. Twenty-four articles were identified that met the inclusion and exclusion criteria. Reported prevalence rates ranged from 10% to 85% with a weighted mean prevalence of 30.6%. The results suggest that university students experience rates of depression that are substantially higher than those found in the general population. Study quality has not improved since 1990.

23260170

Inter-regional cortical thickness correlations are associated with autistic symptoms: a machine-learning approach.

The investigation of neural substrates of autism spectrum disorder using neuroimaging has been the focus of recent literature. In addition, machine-learning approaches have also been used to extract relevant information from neuroimaging data. There are only few studies directly exploring the inter-regional structural relationships to identify and characterize neuropsychiatric disorders. In this study, we concentrate on addressing two issues: (i) a novel approach to extract individual subject features from inter-regional thickness correlations based on structural magnetic resonance imaging (MRI); (ii) using these features in a machine-learning framework to obtain individual subject prediction of a severity scores based on neurobiological criteria rather than behavioral information. In a sample of 82 autistic patients, we have shown that structural covariances among several brain regions are associated with the presence of the autistic symptoms. In addition, we also demonstrated that structural relationships from the left hemisphere are more relevant than the ones from the right. Finally, we identified several brain areas containing relevant information, such as frontal and temporal regions. This study provides evidence for the usefulness of this new tool to characterize neuropsychiatric disorders.

23260169

Effects of promoter methylation on increased expression of polyamine biosynthetic genes in suicide.

Suicide is among the leading causes of death worldwide. The polyamine system has been increasingly implicated in the neurobiology of suicide. Previous research has indicated that epigenetic mechanisms play a role in explaining dysregulation of polyamine genes in suicide completers. Nevertheless, regulatory mechanisms explaining polyamine biosynthetic genes displaying dysregulated expression in suicide completers, including ornithine decarboxylase antizymes 1 and 2 (OAZ1 and OAZ2), S-adenosylmethionine decarboxylase (AMD1), and arginase 2 (ARG2), have yet to be elucidated. In this study, we investigated methylation patterns in the promoter region of OAZ1, OAZ2, AMD1, and ARG2 in Brodmann area 44 from a group of 33 suicide completers and 31 non-suicide controls. We found significant site-specific differences in methylation in the promoter of ARG2 and AMD1 that were also significantly negatively correlated with gene expression. These findings provide further support for a role for the involvement of epigenetic modifications in the regulation of genes associated with polyamine biosynthesis, and which may contribute to the complexity of suicidal behaviors.

23260168

Cloning and expression of feline colony stimulating factor receptor (CSF-1R) and analysis of the species specificity of stimulation by colony stimulating factor-1 (CSF-1) and interleukin-34 (IL-34).

Colony stimulating factor (CSF-1) and its receptor, CSF-1R, have been previously well studied in humans and rodents to dissect the role they play in development of cells of the mononuclear phagocyte system. A second ligand for the CSF-1R, IL-34 has been described in several species. In this study, we have cloned and expressed the feline CSF-1R and examined the responsiveness to CSF-1 and IL-34 from a range of species. The results indicate that pig and human CSF-1 and human IL-34 are equally effective in cats, where both mouse CSF-1 and IL-34 are significantly less active. Recombinant human CSF-1 can be used to generate populations of feline bone marrow and monocyte derived macrophages that can be used to further dissect macrophage-specific gene expression in this species, and to compare it to data derived from mouse, human and pig. These results set the scene for therapeutic use of CSF-1 and IL-34 in cats.

23260167

Emergency department treatments and physiotherapy for acute whiplash: a pragmatic, two-step, randomised controlled trial.

Little is known about the effectiveness of treatments for acute whiplash injury. We aimed to estimate whether training of staff in emergency departments to provide active management consultations was more effective than usual consultations (Step 1) and to estimate whether a physiotherapy package was more effective than one additional physiotherapy advice session in patients with persisting symptoms (Step 2). Step 1 was a pragmatic, cluster randomised trial of 12 NHS Trust hospitals including 15 emergency departments who treated patients with acute whiplash associated disorder of grades I-III. The hospitals were randomised by clusters to either active management or usual care consultations. In Step 2, we used a nested individually randomised trial. Patients were randomly assigned to receive either a package of up to six physiotherapy sessions or a single advice session. Randomisation in Step 2 was stratified by centre. Investigator-masked outcomes were obtained at 4, 8, and 12 months. Masking of clinicians and patients was not possible in all steps of the trial. The primary outcome was the Neck Disability Index (NDI). Analysis was intention to treat, and included an economic evaluation. The study is registered ISRCTN33302125. Recruitment ran from Dec 5, 2005 to Nov 30, 2007. Follow-up was completed on Dec 19, 2008. In Step 1, 12 NHS Trusts were randomised, and 3851 of 6952 eligible patients agreed to participate (1598 patients were assigned to usual care and 2253 patients were assigned to active management). 2704 (70%) of 3851 patients provided data at 12 months. NDI score did not differ between active management and usual care consultations (difference at 12 months 0·5, 95% CI -1·5 to 2·5). In Step 2, 599 patients were randomly assigned to receive either advice (299 patients) or a physiotherapy package (300 patients). 479 (80%) patients provided data at 12 months. The physiotherapy package at 4 months showed a modest benefit compared to advice (NDI difference -3·7, -6·1 to -1·3), but not at 8 or 12 months. Active management consultations and the physiotherapy package were more expensive than usual care and single advice session. No treatment-related serious adverse events or deaths were noted. Provision of active management consultation did not show additional benefit. A package of physiotherapy gave a modest acceleration to early recovery of persisting symptoms but was not cost effective from a UK NHS perspective. Usual consultations in emergency departments and a single physiotherapy advice session for persistent symptoms are recommended. NIHR Health Technology Assessment programme.

23260149

Characterization of full genome of rat hepatitis E virus strain from Vietnam.

We amplified the complete genome of the rat hepatitis E virus (HEV) Vietnam strain (V-105) and analyzed the nucleotide and amino acid sequences. The entire genome of V-105 shared only 76.8%-76.9% nucleotide sequence identities with rat HEV strains from Germany, which suggests that V-105 is a new genotype of rat HEV.

23260147

A molecular roadmap of reprogramming somatic cells into iPS cells.

Factor-induced reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) is inefficient, complicating mechanistic studies. Here, we examined defined intermediate cell populations poised to becoming iPSCs by genome-wide analyses. We show that induced pluripotency elicits two transcriptional waves, which are driven by c-Myc/Klf4 (first wave) and Oct4/Sox2/Klf4 (second wave). Cells that become refractory to reprogramming activate the first but fail to initiate the second transcriptional wave and can be rescued by elevated expression of all four factors. The establishment of bivalent domains occurs gradually after the first wave, whereas changes in DNA methylation take place after the second wave when cells acquire stable pluripotency. This integrative analysis allowed us to identify genes that act as roadblocks during reprogramming and surface markers that further enrich for cells prone to forming iPSCs. Collectively, our data offer new mechanistic insights into the nature and sequence of molecular events inherent to cellular reprogramming.

23260146

Foxa2 and H2A.Z mediate nucleosome depletion during embryonic stem cell differentiation.

Nucleosome occupancy is fundamental for establishing chromatin architecture. However, little is known about the relationship between nucleosome dynamics and initial cell lineage specification. Here, we determine the mechanisms that control global nucleosome dynamics during embryonic stem (ES) cell differentiation into endoderm. Both nucleosome depletion and de novo occupation occur during the differentiation process, with higher overall nucleosome density after differentiation. The variant histone H2A.Z and the winged helix transcription factor Foxa2 both act to regulate nucleosome depletion and gene activation, thus promoting ES cell differentiation, whereas DNA methylation promotes nucleosome occupation and suppresses gene expression. Nucleosome depletion during ES cell differentiation is dependent on Nap1l1-coupled SWI/SNF and INO80 chromatin remodeling complexes. Thus, both epigenetic and genetic regulators cooperate to control nucleosome dynamics during ES cell fate decisions.

23260145

Diabetes risk gene and Wnt effector Tcf7l2/TCF4 controls hepatic response to perinatal and adult metabolic demand.

Most studies on TCF7L2 SNP variants in the pathogenesis of type 2 diabetes (T2D) focus on a role of the encoded transcription factor TCF4 in β cells. Here, a mouse genetics approach shows that removal of TCF4 from β cells does not affect their function, whereas manipulating TCF4 levels in the liver has major effects on metabolism. In Tcf7l2(-/-) mice, the immediate postnatal surge in liver metabolism does not occur. Consequently, pups die due to hypoglycemia. By combining chromatin immunoprecipitation with gene expression profiling, we identify a TCF4-controlled metabolic gene program that is acutely activated in the postnatal liver. In concordance, adult liver-specific Tcf7l2 knockout mice show reduced hepatic glucose production during fasting and display improved glucose homeostasis when maintained on high-fat diet. Furthermore, liver-specific TCF4 overexpression increases hepatic glucose production. These observations imply that TCF4 directly activates metabolic genes and that inhibition of Wnt signaling may be beneficial in metabolic disease.

23260144

Multiple autism-linked genes mediate synapse elimination via proteasomal degradation of a synaptic scaffold PSD-95.

The activity-dependent transcription factor myocyte enhancer factor 2 (MEF2) induces excitatory synapse elimination in mouse neurons, which requires fragile X mental retardation protein (FMRP), an RNA-binding protein implicated in human cognitive dysfunction and autism. We report here that protocadherin 10 (Pcdh10), an autism-spectrum disorders gene, is necessary for this process. MEF2 and FMRP cooperatively regulate the expression of Pcdh10. Upon MEF2 activation, PSD-95 is ubiquitinated by the ubiquitin E3 ligase murine double minute 2 (Mdm2) and then binds to Pcdh10, which links it to the proteasome for degradation. Blockade of the Pcdh10-proteasome interaction inhibits MEF2-induced PSD-95 degradation and synapse elimination. In FMRP-lacking neurons, elevated protein levels of eukaryotic translation elongation factor 1 α (EF1α), an Mdm2-interacting protein and FMRP target mRNA, sequester Mdm2 and prevent MEF2-induced PSD-95 ubiquitination and synapse elimination. Together, our findings reveal roles for multiple autism-linked genes in activity-dependent synapse elimination.

23260143

An endogenous accelerator for viral gene expression confers a fitness advantage.

Many signaling circuits face a fundamental tradeoff between accelerating their response speed while maintaining final levels below a cytotoxic threshold. Here, we describe a transcriptional circuitry that dynamically converts signaling inputs into faster rates without amplifying final equilibrium levels. Using time-lapse microscopy, we find that transcriptional activators accelerate human cytomegalovirus (CMV) gene expression in single cells without amplifying steady-state expression levels, and this acceleration generates a significant replication advantage. We map the accelerator to a highly self-cooperative transcriptional negative-feedback loop (Hill coefficient ∼7) generated by homomultimerization of the virus's essential transactivator protein IE2 at nuclear PML bodies. Eliminating the IE2-accelerator circuit reduces transcriptional strength through mislocalization of incoming viral genomes away from PML bodies and carries a heavy fitness cost. In general, accelerators may provide a mechanism for signal-transduction circuits to respond quickly to external signals without increasing steady-state levels of potentially cytotoxic molecules.

23260142

Crystal structure of the HLA-DM-HLA-DR1 complex defines mechanisms for rapid peptide selection.

HLA-DR molecules bind microbial peptides in an endosomal compartment and present them on the cell surface for CD4 T cell surveillance. HLA-DM plays a critical role in the endosomal peptide selection process. The structure of the HLA-DM-HLA-DR complex shows major rearrangements of the HLA-DR peptide-binding groove. Flipping of a tryptophan away from the HLA-DR1 P1 pocket enables major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket. These conformational changes accelerate peptide dissociation and stabilize the empty HLA-DR peptide-binding groove. Initially, incoming peptides have access to only part of the HLA-DR groove and need to compete with HLA-DR residues for access to the P2 site and the hydrophobic P1 pocket. This energetic barrier creates a rapid and stringent selection process for the highest-affinity binders. Insertion of peptide residues into the P2 and P1 sites reverses the conformational changes, terminating selection through DM dissociation.

23260141

Exosomes mediate stromal mobilization of autocrine Wnt-PCP signaling in breast cancer cell migration.

Stroma in the tumor microenvironment plays a critical role in cancer progression, but how it promotes metastasis is poorly understood. Exosomes are small vesicles secreted by many cell types and enable a potent mode of intercellular communication. Here, we report that fibroblast-secreted exosomes promote breast cancer cell (BCC) protrusive activity and motility via Wnt-planar cell polarity (PCP) signaling. We show that exosome-stimulated BCC protrusions display mutually exclusive localization of the core PCP complexes, Fzd-Dvl and Vangl-Pk. In orthotopic mouse models of breast cancer, coinjection of BCCs with fibroblasts dramatically enhances metastasis that is dependent on PCP signaling in BCCs and the exosome component, Cd81 in fibroblasts. Moreover, we demonstrate that trafficking in BCCs promotes tethering of autocrine Wnt11 to fibroblast-derived exosomes. This work reveals an intercellular communication pathway whereby fibroblast exosomes mobilize autocrine Wnt-PCP signaling to drive BCC invasive behavior.

23260140

MITRAC links mitochondrial protein translocation to respiratory-chain assembly and translational regulation.

Mitochondrial respiratory-chain complexes assemble from subunits of dual genetic origin assisted by specialized assembly factors. Whereas core subunits are translated on mitochondrial ribosomes, others are imported after cytosolic translation. How imported subunits are ushered to assembly intermediates containing mitochondria-encoded subunits is unresolved. Here, we report a comprehensive dissection of early cytochrome c oxidase assembly intermediates containing proteins required for normal mitochondrial translation and reveal assembly factors promoting biogenesis of human respiratory-chain complexes. We find that TIM21, a subunit of the inner-membrane presequence translocase, is also present in the major assembly intermediates containing newly mitochondria-synthesized and imported respiratory-chain subunits, which we term MITRAC complexes. Human TIM21 is dispensable for protein import but required for integration of early-assembling, presequence-containing subunits into respiratory-chain intermediates. We establish an unexpected molecular link between the TIM23 transport machinery and assembly of respiratory-chain complexes that regulate mitochondrial protein synthesis in response to their assembly state.

23260139

Force fluctuations within focal adhesions mediate ECM-rigidity sensing to guide directed cell migration.

Cell migration toward areas of higher extracellular matrix (ECM) rigidity via a process called "durotaxis" is thought to contribute to development, immune response, and cancer metastasis. To understand how cells sample ECM rigidity to guide durotaxis, we characterized cell-generated forces on the nanoscale within single mature integrin-based focal adhesions (FAs). We found that individual FAs act autonomously, exhibiting either stable or dynamically fluctuating ("tugging") traction. We show that a FAK/phosphopaxillin/vinculin pathway is essential for high FA traction and to enable tugging FA traction over a broad range of ECM rigidities. We show that tugging FA traction is dispensable for FA maturation, chemotaxis, and haptotaxis but is critical to direct cell migration toward rigid ECM. We conclude that individual FAs dynamically sample rigidity by applying fluctuating pulling forces to the ECM to act as sensors to guide durotaxis, and that FAK/phosphopaxillin/vinculin signaling defines the rigidity range over which this dynamic sensing process operates.

23260138

CapSeq and CIP-TAP identify Pol II start sites and reveal capped small RNAs as C. elegans piRNA precursors.

Piwi-interacting (pi) RNAs are germline-expressed small RNAs linked to epigenetic programming. C. elegans piRNAs are thought to be transcribed as independent gene-like loci. To test this idea and to identify potential transcription start (TS) sites for piRNA precursors, we developed CapSeq, an efficient enzymatic method for 5' anchored RNA profiling. Using CapSeq, we identify candidate TS sites, defined by 70-90 nt sequence tags, for >50% of annotated Pol II loci. Surprisingly, however, these CapSeq tags failed to identify the overwhelming majority of piRNA loci. Instead, we show that the likely piRNA precursors are ∼26 nt capped small (cs) RNAs that initiate precisely 2 nt upstream of mature piRNAs and that piRNA processing or stability requires a U at the csRNA +3 position. Finally, we identify a heretofore unrecognized class of piRNAs processed from csRNAs that are expressed at promoters genome wide, nearly doubling the number of piRNAs available for genome surveillance.

23260137

An inhibitor of nonhomologous end-joining abrogates double-strand break repair and impedes cancer progression.

DNA Ligase IV is responsible for sealing of double-strand breaks (DSBs) during nonhomologous end-joining (NHEJ). Inhibiting Ligase IV could result in amassing of DSBs, thereby serving as a strategy toward treatment of cancer. Here, we identify a molecule, SCR7 that inhibits joining of DSBs in cell-free repair system. SCR7 blocks Ligase IV-mediated joining by interfering with its DNA binding but not that of T4 DNA Ligase or Ligase I. SCR7 inhibits NHEJ in a Ligase IV-dependent manner within cells, and activates the intrinsic apoptotic pathway. More importantly, SCR7 impedes tumor progression in mouse models and when coadministered with DSB-inducing therapeutic modalities enhances their sensitivity significantly. This inhibitor to target NHEJ offers a strategy toward the treatment of cancer and improvement of existing regimens.

23260136

Whole-genome sequencing in autism identifies hot spots for de novo germline mutation.

De novo mutation plays an important role in autism spectrum disorders (ASDs). Notably, pathogenic copy number variants (CNVs) are characterized by high mutation rates. We hypothesize that hypermutability is a property of ASD genes and may also include nucleotide-substitution hot spots. We investigated global patterns of germline mutation by whole-genome sequencing of monozygotic twins concordant for ASD and their parents. Mutation rates varied widely throughout the genome (by 100-fold) and could be explained by intrinsic characteristics of DNA sequence and chromatin structure. Dense clusters of mutations within individual genomes were attributable to compound mutation or gene conversion. Hypermutability was a characteristic of genes involved in ASD and other diseases. In addition, genes impacted by mutations in this study were associated with ASD in independent exome-sequencing data sets. Our findings suggest that regional hypermutation is a significant factor shaping patterns of genetic variation and disease risk in humans.

23260135

MeCP2 binds to 5hmC enriched within active genes and accessible chromatin in the nervous system.

The high level of 5-hydroxymethylcytosine (5hmC) present in neuronal genomes suggests that mechanisms interpreting 5hmC in the CNS may differ from those present in embryonic stem cells. Here, we present quantitative, genome-wide analysis of 5hmC, 5-methylcytosine (5mC), and gene expression in differentiated CNS cell types in vivo. We report that 5hmC is enriched in active genes and that, surprisingly, strong depletion of 5mC is observed over these regions. The contribution of these epigenetic marks to gene expression depends critically on cell type. We identify methyl-CpG-binding protein 2 (MeCP2) as the major 5hmC-binding protein in the brain and demonstrate that MeCP2 binds 5hmC- and 5mC-containing DNA with similar high affinities. The Rett-syndrome-causing mutation R133C preferentially inhibits 5hmC binding. These findings support a model in which 5hmC and MeCP2 constitute a cell-specific epigenetic mechanism for regulation of chromatin structure and gene expression.

23260134

The origin of membrane bioenergetics.

Harnessing energy as ion gradients across membranes is as universal as the genetic code. We leverage new insights into anaerobe metabolism to propose geochemical origins that account for the ubiquity of chemiosmotic coupling, and Na(+)/H(+) transporters in particular. Natural proton gradients acting across thin FeS walls within alkaline hydrothermal vents could drive carbon assimilation, leading to the emergence of protocells within vent pores. Protocell membranes that were initially leaky would eventually become less permeable, forcing cells dependent on natural H(+) gradients to pump Na(+) ions. Our hypothesis accounts for the Na(+)/H(+) promiscuity of bioenergetic proteins, as well as the deep divergence between bacteria and archaea.

23260133

A dimer to bridge early autophagosomal membranes.

The Atg1/ULK complex plays a key role in the early stages of autophagosome assembly. In this issue, Ragusa et al. reveal the molecular basis for some interactions within this complex, finding that the crescent-shaped Atg17 dimer is critical for autophagy, whereas Atg1 may have the ability to cluster membranes.

23260132

Blending hippo and WNT: sharing messengers and regulation.

Two new studies reveal ways in which the Wnt pathway commandeers Hippo components for signaling. Azzolin et al. show how the Hippo transcription factor TAZ mediates Wnt signals, and Rosenbluh et al. show how β-catenin and YAP1 form a kinase-regulated complex with transcription factor TBX5.

23260131

Loaded dice for human genome mutation.

Not all bases in the human genome are equally prone to chance mutations. Michaelson et al. show that individuals can acquire clusters of de novo DNA changes and propose that the likelihood of single base pair change differs across three orders of magnitude.

23260129

There and back again: new single-molecule insights in the motion of DNA repair proteins.

Cellular DNA repair machines are constantly at work supporting the integrity of our genomes. Numerous proteins cooperate to form a complex and adaptive system dedicated to detection and timely processing of DNA damage. The molecular underpinnings of how these proteins locate and discriminate DNA lesions, match homologous sequences, mend the DNA and attend to a replication in distress are of a paramount biomedical importance, but in many cases remain unclear. Combined with more conventional tools, single-molecule biochemistry has been stepping in to address the age-old problems in the DNA repair field. This review will address new insights into diffusive properties of three DNA repair systems: I will discuss the emerging model of how MutS homologues locate and respond to mismatches in the dsDNA; the mechanism by which RAD52 promotes annealing of complementary DNA strands coated with ssDNA binding protein RPA; and how the nucleoprotein filament formed by RecA recombinase on ssDNA searches for homology within duplex DNA. These three distinct DNA repair factors exemplify the dynamic nature of cellular DNA repair machines revealed by single-molecule studies.

23260128

Worldwide burden of HIV in transgender women: a systematic review and meta-analysis.

Previous systematic reviews have identified a high prevalence of HIV infection in transgender women in the USA and in those who sell sex (compared with both female and male sex workers). However, little is known about the burden of HIV infection in transgender women worldwide. We aimed to better assess the relative HIV burden in all transgender women worldwide. We did a systematic review and meta-analysis of studies that assessed HIV infection burdens in transgender women that were published between Jan 1, 2000, and Nov 30, 2011. Meta-analysis was completed with the Mantel-Haenszel method, and random-effects modelling was used to compare HIV burdens in transgender women with that in adults in the countries for which data were available. Data were only available for countries with male-predominant HIV epidemics, which included the USA, six Asia-Pacific countries, five in Latin America, and three in Europe. The pooled HIV prevalence was 19·1% (95% CI 17·4-20·7) in 11 066 transgender women worldwide. In 7197 transgender women sampled in ten low-income and middle-income countries, HIV prevalence was 17·7% (95% CI 15·6-19·8). In 3869 transgender women sampled in five high-income countries, HIV prevalence was 21·6% (95% CI 18·8-24·3). The odds ratio for being infected with HIV in transgender women compared with all adults of reproductive age across the 15 countries was 48·8 (95% CI 21·2-76·3) and did not differ for those in low-income and middle-income countries compared with those in high-income countries. Our findings suggest that transgender women are a very high burden population for HIV and are in urgent need of prevention, treatment, and care services. The meta-analysis showed remarkable consistency and severity of the HIV disease burden among transgender women. Center for AIDS Research at Johns Hopkins and the Center for Public Health and Human Rights at the JHU Bloomberg School of Public Health.

23260125

Psychological and social adjustment in older transsexual people.

Several forces conspire to make the later decades a climacteric for transgendered persons. This paper will examine the social, emotional, and hormonal influences that entwine and challenge the stability of the elderly transgendered person. Case studies and therapeutic interventions will be addressed. Within our article, the cases are fictitious and therefore consent was not required for our vignettes.

23260124

Calcyclin binding protein and Siah-1 interacting protein in Alzheimer's disease pathology: neuronal localization and possible function.

The calcyclin binding protein and Siah-1 interacting protein (CacyBP/SIP) protein was shown to play a role in the organization of microtubules. In this work we have examined the neuronal distribution and possible function of CacyBP/SIP in cytoskeletal pathophysiology. We have used brain tissue from Alzheimer's disease (AD) patients and from transgenic mice modeling 2 different pathologies characteristic for AD: amyloid and tau. In the brain from AD patients, CacyBP/SIP was found to be almost exclusively present in neuronal somata, and in control patients it was seen in the somata and neuronal processes. In mice doubly transgenic for amyloid precursor protein and presenilin 1 there was no difference in CacyBP/SIP neuronal localization in comparison with the nontransgenic animals. By contrast in tau transgenic mice, localization of CacyBP/SIP was similar to that observed for AD patients. To find the relation between CacyBP/SIP and tau we examined dephosphorylation of tau by CacyBP/SIP. We found that indeed it exhibited phosphatase activity toward tau. Altogether, our results suggest that CacyBP/SIP might play a role in AD pathology.

23260123

The relationship among internal resilience, smoking, alcohol use, and depression symptoms in emerging adults transitioning out of child welfare.

It is well established that child maltreatment reflects a context of risk for multiple negative outcomes. Identifying factors that protect against negative outcomes is important for the development of strengths-based approaches that emphasize resilience, particularly for youth transitioning out of the child welfare system. The current study examined the relationship between an internal resilience measure, the Connor-Davidson Resilience Scale (CD-RISC; Connor & Davidson, 2003), and several external measures of resilience and behavioral outcomes (tobacco use and dependence, alcohol use and problems, and depression symptoms). In addition, two models of resilience were examined in the context of child maltreatment: a compensatory model and a risk-protection model. Ninety-three emerging adults (ages 18-25) who were making the transition out of child welfare completed self-report measures of child maltreatment, internal resilience (CD-RISC), external resilience (academic achievement, religious and community involvement, monitoring by caregivers, and presence of an adult mentor), alcohol and tobacco use, and depression symptoms. Internal resilience was significantly associated with involvement in religion and community, and monitoring by caregivers. In addition, internal resilience was negatively associated with past year smoking and nicotine dependence, and with symptoms of depression. Hierarchical regression analyses were conducted to examine the direct and interaction effects of resilience on depression symptoms in the context of child maltreatment. When internal resilience was added to the model, it made a significant contribution to depression scores over and above child maltreatment (physical, sexual, and emotional abuse; emotional neglect). In addition, there was a significant Sexual Abuse×Resilience interaction, wherein high resilience was associated with a reduction in depression scores at higher levels of sexual abuse. These findings support internal resilience as both a compensatory and protective factor for depression symptoms in the context of sexual abuse among emerging adults transitioning out of child welfare. Prevention and early intervention within child welfare should include strengthening internal resilience, with continued monitoring of competencies through the transition from adolescence to emerging adulthood.

23260122

Child maltreatment and repeat presentations to the emergency department for suicide-related behaviors.

To identify factors associated with repeat emergency department (ED) presentations for suicide-related behaviors (SRB) - hereafter referred to as repetition - among children/youth to aid secondary prevention initiatives. To compare rates of repetition in children/youth with substantiated maltreatment requiring removal from their parental home with their peers in the general population. A population-based (retrospective) cohort study was established for children/youth with a first ED SRB presentation at risk for repetition in the Province of Ontario, Canada between 1 January 2004 and 31 December 2008. Children/youth legally removed from their parental home because of substantiated maltreatment (n=179) and their population-based peers (n=6,305) were individually linked to administrative health care records over time to ascertain social, demographic, and clinical information and subsequent ED presentations for SRB during follow-up. These children/youth were described and their repetition-free probabilities over time compared. To identify factors associated with repetition we fit multivariable, recurrent event survival analysis models stratified by repetition and present unadjusted and adjusted hazard ratios (HRs) and 95% confidence intervals (CIs). Children/youth with substantiated maltreatment (as noted) were two times more likely to have repetition than their peers after adjustments for social, demographic, and clinical factors (conditional on prior ED SRB presentations). A number of these factors were independently associated with repetition. No one factor distinguished between having a first and second repetition nor was more strongly associated with repetition than another. The risk of repetition is higher in children with substantiated maltreatment (as noted) than their peers. No one factor stood out as predictive of repetition. Implications for secondary prevention initiatives include a non-selective approach, sensitive to family difficulties and the need to better contextualize repetition and harness data linkages.

23260121

Placement decisions and disparities among Aboriginal children: further analysis of the Canadian incidence study of reported child abuse and neglect part A: comparisons of the 1998 and 2003 surveys.

Fluke et al. (2010) analyzed Canadian Incidence Study on Reported Child Abuse and Neglect (CIS) data collected in 1998 to explore the influence of clinical and organizational characteristics on the decision to place Aboriginal children in an out-of-home placement at the conclusion of a child maltreatment investigation. This study explores this same question using CIS data collected in 2003 which included a larger sample of Aboriginal children and First Nations child and family service agencies. The decision to place a child in an out-of-home placement was examined using data from the Canadian Incidence Study of Reported Child Abuse and Neglect-2003 and a reanalysis of CIS-1998 data (Fluke et al., 2010). The CIS-2003 dataset includes information on nearly 12,000 child maltreatment investigations from the time of report to case disposition. The CIS-2003 also captures information on the characteristics of investigating workers and the child welfare organizations for which they work. Multi-level statistical models were developed to analyze the influence of clinical and organizational variables using MPlus software. MPlus allows the use of dichotomous outcome variables, which are more reflective of decision-making in child welfare and facilitates the specific case of the logistic link function for binary outcome variables under maximum likelihood estimation. Final models revealed the proportion of investigations conducted by the child welfare agency involving Aboriginal children was a key single agency level predictor of the placement decision. Specifically, the higher the proportion of investigations of Aboriginal children, the more likely placement was to occur. Contrary to the findings in the first paper (Fluke et al., 2010), individual Aboriginal status also remained significant in the final model at the first level. Further analysis needs to be conducted to further understand individual and organizational level variables that may influence decisions regarding placement of Aboriginal children. There is also a need for research that is sensitive to differences among, and between, Métis, First Nations and Inuit communities. Results are not generalizable to Québec because data from this province were excluded.

23260120

Common elements for the psychotherapeutic management of patients with self injurious behavior.

Current research suggests that effective psychotherapies for Self Injurious Behavior (SIB) in the context of Borderline Personality Disorder (BPD) contain generic common elements which are responsible for their success. Because of the links between BPD, SIB, and child abuse, it is likely that these common elements can also be applied to the psychotherapy of survivors of child abuse who engage in SIB. This article will reviews several common elements which recent literature has suggested are important and suggest techniques by which community practitioners can incorporate these elements into their practice with adult survivors of child abuse who engage in SIB. This article summarizes recent key articles on common elements in psychotherapy for BPD. It refers to the treatment manuals for the specialized psychotherapies as well as General Psychiatric Management (GPM), a novel and more accessible approach, to further elaborate on these common elements and their potential usefulness in the community. This paper identifies seven common elements which appear both in the recent literature and in GPM: a coherent model to understand SIB, an active therapeutic stance, validation balanced with change-oriented techniques, encouragement of self-agency, establishment of a connection between actions and feelings, a method for assessing lethality, and access to supervision. By referring to current treatment strategies for BPD, this paper suggests methods by which these common elements can be incorporated into practice by community practitioners working with adult survivors of child abuse who engage in SIB. Despite a lack of access to intensive training or the resources required for multimodal treatments, there are many strategies from established treatments for BPD which can be used by community practitioners to address SIB. Because treatments for BPD often focus on addressing SIB, and because adult survivors of child abuse often engage in SIB, whether or not they have BPD, it is likely that these techniques are applicable to practice with adult survivors. Further research is, however, necessary to empirically examine these common elements and their potential utility.

23260119

Patterns of service use, individual and contextual risk factors, and resilience among adolescents using multiple psychosocial services.

Very little research has examined the relationship between resilience, risk, and the service use patterns of adolescents with complex needs who use multiple formal and mandated services such as child welfare, mental health, juvenile justice, and special educational supports. This article reports on a study of 497 adolescents in Atlantic Canada who were known to have used at least 2 of these services in the last 6 months. It was hypothesized that greater service use and satisfaction with services would predict both resilience, and better functional outcomes such as prosocial behavior, school engagement and participation in community. Youth who were known to be multiple service users and who were between the ages of 13 and 21 participated in the study. Participants completed a self-report questionnaire administered individually. Path analysis was used to determine the relationship between risk, service use, resilience, and functional outcomes. MANOVA was then used to determine patterns of service use and service use satisfaction among participants. Findings show that there was no significant relationship between service use history and resilience or any of the three functional outcomes. Service use satisfaction, a measure of an adolescent's perception of the quality of the services received, did however show a strong positive relationship with resilience. Resilience mediates the impact of risk factors on outcomes and is affected positively by the quality, but not the quantity, of the psychosocial services provided to adolescents with complex needs. Results show that resilience is related to service satisfaction but not the quantity of services used by youth. Coordinated services may not increase resilience or be more effective unless the quality of individual services is experienced by an adolescent receiving intervention as personally empowering and sensitive to his or her needs.

23260118

Two decades later: The resilience and post-traumatic responses of Indigenous Quechua girls and adolescents in the aftermath of the Peruvian armed conflict.

In comparison to other traumatic events, the impact of a childhood during war on resilience later in life has been seldom examined. The aim of this study was therefore to examine the long term outcomes of post-traumatic responses and resilience of a sample of adult Indigenous Quechua women, who were girls or adolescents during the Peruvian armed conflict (1980-1995). The study instruments (Harvard trauma questionnaire part I and IV; Connor-Davidson resilience scale; life stress questionnaire) were translated to Quechua and cross-culturally validated. A cross sectional survey design was used in 2010 to collect data from a convenience sample of 75 participants (25-45 years old) in Ayacucho, Peru, the region most affected by the conflict. Data was examined using hierarchical regression analyses. Participants reported extreme exposure to violence (e.g., sexual violence, torture, combat, death of family members, and forced displacement) during the armed conflict, but surprisingly, only 5.3% reported a current level of symptoms that may indicate a possible post-traumatic stress disorder (PTSD). Resilience scores and number of years exposed to conflict as a child were not associated with PTSD symptoms; instead only the degree of exposure to violence, and current level of stress contributed to the variance of PTSD-related symptoms. Conversely, resilience and current stress contributed to the variance of trauma symptoms when measured by local idioms of distress. Findings should be interpreted with caution, due to limitations in the content validity of instruments, risk of inaccurate recall, use of individual explanations of distress (such as PTSD) for collective experiences of violence, use of non-indigenous frameworks to examine Indigenous resilience, and other methodological concerns. The study however highlights the high degree of traumatic exposure of these former war children. While the prevalence of potential PTSD was astonishingly low in this sample, a number of women still suffer from significant distress two decades after the traumatic events. Therefore, post-conflict interventions should renew efforts to foster the resilience of marginalized populations disproportionately targeted by violence and advocate for enhanced protection of women and children in current armed conflicts.

23260117

Finding the way out: a non-dichotomous understanding of violence and depression resilience of adolescents who are exposed to family violence.

In this cross-sectional study on family violence and resilience in a random sample of 5,149 middle school students with a mean age of 14.5 years from four EU-countries (Austria, Germany, Slovenia, and Spain) we examined the prevalence of exposure to family violence, and we worked from the premise that adolescent can be resilient to family violence. We expanded the definition of resilience to include the absence of both physical aggression and depression symptoms in adolescents who have been exposed to violence in their families and extended our understanding of resilience to include three levels which we describe as: "resilient", "near-resilient" and "non-resilient", thus responding to calls for a more fluid and paths-based understanding of resilience. Data were collected via self-administered surveys consisting of a number of subscales that investigate depression symptoms and physical aggression. The study was analyzed with a three-stage strategy using logistic regression procedures, in which regression analyses were conducted separately for girls and boys using seven steps for modeling the three resilience levels. More than 30% of our respondents reported experiencing family violence. Contrary to previous research findings, our data showed that structural characteristics like country, gender, socio-economic status and migration status were minimally predictive of violence and depression resilience at any level. Overall, for both sexes, despite some small but significant sex differences, resilience is strongly linked to personal and relational characteristics and the absence of experiences that involved exposure to and direct experiences with violence. Resilience supportive factors confirmed by this study are: higher emotional self-control, talking with parents or friends about violence, seeking help to avoid violence, and not endorsing aggression supportive beliefs. Also key to resilience are irrespective of country, gender, and SES are lower levels of experience with: victimization by boys, engagement in physical altercation with boys, parental abuse, witnessing of physical spousal abuse, exposure to an authoritarian (harsh) parenting style and verbal aggression from teachers. From a content perspective this means that resilience is more than the absence of one or two behavioral factors. This also means that positive changes in resilience levels can be facilitated by supporting constructive personal and social relationships with family members, peers, and teachers. These results are discussed in terms of their practical implications for policy and intervention.

23260116

Administrative data linkage as a tool for child maltreatment research.

Linking administrative data records for the same individuals across services and over time offers a powerful, population-wide resource for child maltreatment research that can be used to identify risk and protective factors and to examine outcomes. Multistage de-identification processes have been developed to protect privacy and maintain confidentiality of the datasets. Lack of information on those not coming to the attention of child protection agencies, and limited information on certain variables, such as individual-level SES and parenting practices, is outweighed by strengths that include large and unbiased samples, objective measures, comprehensive long-term follow-up, continuous data collection, and relatively low expense. Ever emerging methodologies and expanded holdings ensure that research using linked population-wide databases will make important contributions to the study of child maltreatment.

23260115

Understanding risk and protective factors for child maltreatment: the value of integrated, population-based data.

In this article, we argue for expanded efforts to integrate administrative data systems as a "practical strategy" for developing a richer understanding of child abuse and neglect. Although the study of child maltreatment is often critiqued for being atheoretical, we believe that a more pressing concern is the absence of population-based and prospective epidemiological data that can be used to better understand the distribution and interacting nature of risk and protective factors for maltreatment. We begin by briefly addressing the relevance of empirical observations to etiological theories of child maltreatment. Although the latter is widely cited as critical to the development of effective prevention and intervention responses, less attention has been paid to the role of population-based data in the development of theories relevant to highly applied research questions such as those pertaining to child abuse and neglect. We then discuss how child protection data, in isolation, translates into a relatively narrow range of questions that can be asked and answered, with an inherently pathology-focused construction of risks and little attention paid to strengths or protective factors. We next turn to examples of recent findings--spanning multiple countries--emerging from information integrated across data systems, concluding by calling for expanded administrative data linkages in an effort to better understand and prevent child maltreatment.

23260114

Resilience after maltreatment: the importance of social services as facilitators of positive adaptation.

This practice note will show that resilience among children who have been maltreated is the result of multiple protective factors, including the quality of the services provided to children exposed to chronic adversity. This social ecological perspective of resilience suggests that resilience is a process resulting from interactions between individuals and their environments, and depends upon individual characteristics (temperament and personality), the social determinants of health that affect children and children's families, formal interventions by multiple service providers (child welfare, special education, mental health, addictions, public health, and juvenile corrections), and the social policies that influence service provision to vulnerable populations. Clinicians and researchers concerned with the resilience of chronically abused and neglected children have tended to overlook the protective processes unique to children who have been abused that are different from the protective processes that promote positive development among children who have experienced no maltreatment. Most importantly, studies of resilience among maltreated children have rarely investigated the impact child welfare interventions have on the resilience of children who have been maltreated, mistakenly attributing children's abilities to cope to be the result of individual factors rather than the responsiveness of service providers and governments to tailor interventions to children's needs. To enhance the likelihood of resilience among maltreated children, those who design and implement interventions need to address three aspects of resilience-related programming: make social supports and formal services more available and accessibility; design programs flexibly so that they can respond to the differential impact specific types of interventions have on children who are exposed to different forms of maltreatment; and design interventions to be more focused on subpopulations of children who have experienced maltreatment rather than diffuse population-wide initiatives.

23260113

Psychometric properties of the child and youth resilience measure (CYRM-28) among samples of French Canadian youth.

Explore the psychometric properties of the French Canadian version of the Child and Youth Resilience Measure (CYRM-28, Resilience Research Center, 2009; Ungar et al., 2008) in youth samples. Two investigations were conducted. Participants in Study 1 were 589 youth (60% female) in grades 10-12 from 2 urban public high schools. Participants in Study 2 were 246 youth (48% female) from a rural public high school, 28% from First Nations. All participants completed the French CYRM-28 and measures of self-esteem and self-acceptance/mindfulness. Participants in Study 2 completed additional measures evaluating their sense of empowerment, trauma symptoms, family problems, and relationship with parents. Factor analysis identified three components correlated to each other: individual, family and community resilience. Evidence provides initial support for the construct validity of the scale by correlations with measures of self-esteem, self-acceptance/mindfulness, empowerment, trauma symptoms, relationship with parents and differences according to gender and a history of sexual abuse. The present results, the first to explore the psychometric properties of the French version of the CYRM-28, provide preliminary data supporting the reliability and validity of a global scale including 27 items. However, our results reveal a different factorial structure compared to previous studies using the CYRM-28. Future studies are needed to further document the validity of the scale.

23260112

The influence of unemployment and divorce rate on child help-seeking behavior about violence, relationships, and other issues.

This study examined the influence of community unemployment and divorce rate on child help-seeking behavior about violence and relationships via a telephone and Internet helpline. Time series analysis was conducted on monthly call volumes to a child helpline ('De Kindertelefoon') in the Netherlands from 2003 to 2008 and on the topics discussed (primarily Violence and Relationships) from 1994 to 2008 in answered calls and chats. As unemployment rises, the number of calls to the helpline increases. With increased unemployment, the share of conversations about violence and about relationships is found to be higher. When the divorce rate goes up, the number of calls to the helpline increases, but the share of Violence and of Relationships decreases. In addition, the share of contacts about Violence is increasing over time, while the share about Relationships is decreasing. Furthermore, the showing of violent movies is associated with fewer attempted helpline calls. Finally, seasonal variation in call volumes and the shares of calls about Violence and Relationships are found. Our data provide a unique and direct perspective on child help-seeking behavior. Our results suggest that more resources need to be devoted to resolving children's problems during times of unemployment as they seek more help particularly with respect to violence. Increased training of counselors with regard to children reporting incidents of violence is particularly important, as the share of contacts about violence is increasing over time and during a recession. Resources are especially needed as, on average, less than half of the calls to child helplines are actually answered.

23260110

Interleukin 2-inducible T cell kinase (ITK) facilitates efficient egress of HIV-1 by coordinating Gag distribution and actin organization.

Interleukin 2-inducible T cell kinase (ITK) influences T cell signaling by coordinating actin polymerization and polarization as well as recruitment of kinases and adapter proteins. ITK regulates multiple steps of HIV-1 replication, including virion assembly and release. Fluorescent microscopy was used to examine the functional interactions between ITK and HIV-1 Gag during viral particle release. ITK and Gag colocalized at the plasma membrane and were concentrated at sites of F-actin accumulation and membrane lipid rafts in HIV-1 infected T cells. There was polarized staining of ITK, Gag, and actin towards sites of T cell conjugates. Small molecule inhibitors of ITK disrupted F-actin capping, perturbed Gag-ITK colocalization, inhibited virus like particle release, and reduced HIV replication in primary human CD4+ T cells. These data provide insight as to how ITK influences HIV-1 replication and suggest that targeting host factors that regulate HIV-1 egress provides an innovative strategy for controlling HIV infection.

23260109

Intracellular nucleotide levels and the control of retroviral infections.

Retroviruses consume cellular deoxynucleoside triphosphates (dNTPs) to convert their RNA genomes into proviral DNA through reverse transcription. While all retroviruses replicate in dividing cells, lentiviruses uniquely replicate in nondividing cells such as macrophages. Importantly, dNTP levels in nondividing cells are extremely low, compared to dividing cells. Indeed, a recently discovered anti-HIV/SIV restriction factor, SAMHD1, which is a dNTP triphosphohydrolase, is responsible for the limited dNTP pool of nondividing cells. Lentiviral reverse transcriptases (RT) uniquely stay functional even at the low dNTP concentrations in nondividing cells. Interestingly, Vpx of HIV-2/SIVsm proteosomally degrades SAMHD1, which elevates cellular dNTP pools and accelerates lentiviral replication in nondividing cells. These Vpx-encoding lentiviruses rapidly replicate in nondividing cells by encoding both highly functional RTs and Vpx. Here, we discuss a series of mechanistic and virological studies that have contributed to conceptually linking cellular dNTP levels and the adaptation of lentiviral replication in nondividing cells.

23260111

An AUG codon upstream of rev and env open reading frames ensures optimal translation of the simian immunodeficiency virus Env protein.

The mRNAs encoding the Rev and Env proteins of simian immunodeficiency virus (SIV) are unique because upstream translation start codons are present that may modulate the expression of these viral proteins. We previously reported the regulatory effect of a small upstream open reading frame (ORF) on Rev and Env translation. Here we study this mechanism in further detail by modulating the strength of the translation signals upstream of the open reading frames in subgenomic reporters. Furthermore, the effects of these mutations on SIV gene expression and viral replication are analyzed. An intricate regulatory mechanism is disclosed that allows the virus to express a balanced amount of these two proteins.

23260108

Cell-cell transmission allows human T-lymphotropic virus 1 to circumvent tetherin restriction.

Tetherin is part of the cellular innate immunity and impedes cell-free transmission of viruses that bud from the plasma membrane by retaining them on the cell surface. Some viruses have evolved activities in different proteins such as Vpu (HIV-1), K-protein (KSHV), Nef (SIV) or Env (HIV-2) to downregulate tetherin and overcome its restriction. We found that chronically HTLV-1 infected T-cell lines express eightfold more tetherin than uninfected transformed T-cell lines suggesting that tetherin expression is not inhibited by the virus. We observed that even small amounts of exogenous tetherin caused the retention of HTLV-1 on the cell surface and severely reduced cell-free infectivity of HTLV-1, but that cell-cell transmission, which is more relevant for HTLV-1, was significantly less decreased. However, knock-down of tetherin expresssion resulted in a slight increase in cell-cell infection indicating that the protein does not enhance this route of transmission.

23260107

Dog nectin-4 is an epithelial cell receptor for canine distemper virus that facilitates virus entry and syncytia formation.

Canine distemper virus (CDV) was shown to use dog nectin-4 as a receptor to gain entry into epithelial cells. RNA from dog placenta or MDCK kidney cells was isolated and cDNAs were prepared. Two splice variants of dog nectin-4 were identified. A deletion of 25 amino acids was found in the cytoplasmic domain of dog nectin-4 from MDCK cells, corresponding to a splice variant that is also seen in murine nectin-4, and did not affect its role as a receptor. Both dog nectin-4 and human nectin-4 could function as an entry factor for CDV containing an EGFP reporter gene. Inhibition of dog nectin-4 expression by RNAi or nectin-4 antibodies decreased CDV titers and EGFP fluorescence. Finally, dog nectin-4 also promotes syncytia formation, which could be inhibited by siRNA treatment. These data confirm that dog nectin-4 can be used by CDV to gain entry into epithelial cells, and facilitate virus spread.

23260106

Birth weight and carotid artery intima-media thickness.

To determine the association between birth weight and carotid artery intima-media thickness (CIMT), a measure of atherogenesis, in a population of 11-year-old children. CIMT measured by high-resolution ultrasound, and birth registry data were available for 670 children of the Southern California Children's Health Study. Multivariate regression analyses were performed to investigate the association between birth weight and CIMT, with adjustment for child's health status and lifestyle, pregnancy information, and parental health. Mean CIMT was 0.57 mm (SD 0.04). We found a nonlinear association between birth weight and CIMT, with an increase in CIMT of 0.014 mm in the fifth (P value .01) compared with the third birth weight quintile. These associations were robust in subsample analyses in children considered normal-weight by gestational age or in term-born children. No significant association with CIMT was found for the lowest quintile. Greater birth weight was significantly associated with increased CIMT at age 11 years. No evidence for an impact of lower birth weight was found. The predictive value of childhood CIMT on future cardiovascular outcomes is largely unknown, but strong associations between childhood cardiovascular disease risk factors and adult vascular disease suggest that increased CIMT in childhood may be clinically important.

23260104

Alterations in ventricular structure and function in obese adolescents with nonalcoholic fatty liver disease.

To determine the association among nonalcoholic fatty liver disease (NAFLD), metabolic function, and cardiac function in obese adolescents. Intrahepatic triglyceride (IHTG) content (magnetic resonance spectroscopy), insulin sensitivity and β-cell function (5-hour oral glucose tolerance test with mathematical modeling), and left ventricular function (speckle tracking echocardiography) were determined in 3 groups of age, sex, and Tanner matched adolescents: (1) lean (n=14, body mass index [BMI]=20±2 kg/m2); (2) obese with normal (2.5%) IHTG content (n=15, BMI=35±3 kg/m2); and (3) obese with increased (8.7%) IHTG content (n=15, BMI=37±6 kg/m2). The disposition index (β-cell function) and insulin sensitivity index were ∼45% and ∼70% lower, respectively, and whole body insulin resistance, calculated by homeostasis model of assessment-insulin resistance (HOMA-IR), was ∼60% greater, in obese than in lean subjects, and ∼30% and ∼50% lower and ∼150% greater, respectively, in obese subjects with NAFLD than those without NAFLD (P<.05 for all). Left ventricular global longitudinal systolic strain and early diastolic strain rates were significantly decreased in obese than in lean subjects, and in obese subjects with NAFLD than those without NAFLD (P<.05 for all), and were independently associated with HOMA-IR (β=0.634). IHTG content was the only significant independent determinant of insulin sensitivity index (β=-0.770), disposition index (β=-0.651), and HOMA-IR (β=0.738). These findings demonstrate that the presence of NAFLD in otherwise asymptomatic obese adolescents is an early marker of cardiac dysfunction.

23260103

Esophageal mechanosensitive mechanisms are impaired in neonates with hypoxic-ischemic encephalopathy.

To test the hypothesis that esophageal mechanodistention in infants with hypoxic-ischemic encephalopathy (HIE) results in altered upper esophageal sphincter (UES), esophageal body, and lower esophageal sphincter (LES) responses, compared with controls. As a secondary aim, we tested the hypothesis that infants with HIE receiving therapeutic hypothermia had different aerodigestive reflex characteristics than infants with HIE who received traditional neonatal care. Provocative esophageal manometry was performed in 34 neonates (27 with HIE and 7 controls). Mechanodistention was performed using graded volumes of air. Peristaltic reflexes, UES contractile reflexes, and LES relaxation reflexes were analyzed for frequency, magnitude, and aberrancies. Infants with HIE demonstrated more rapid recruitment of responses and greater UES magnitude (P < .05). They had more frequent secondary peristalsis and lower LES nadir pressures with prolonged LES nadir durations (P < .05). Most notable were the prolonged peristaltic response durations and increases in the number of polymorphic waveforms (P < .05). Compared with infants with HIE receiving traditional care, infants with HIE treated with hypothermia had higher UES pressures and shorter peristaltic response duration (P < .05). Mechanodistention in infants with HIE results in upregulation of central vagal effects (ie, heightened cholinergic excitatory responses as demonstrated by exaggerated UES contractile reflex activity and heightened inhibitory responses evident by exaggerated LES relaxation reflex activity). Prolonged and poorly coordinated peristaltic responses may underlie dysfunction of aerodigestive regulation. Modulation of sensorimotor aspects of aerodigestive reflexes is altered in infants with HIE, and hypothermia may further modify such effects.

23260102

Arteriopathy, D-dimer, and risk of poor neurologic outcome in childhood-onset arterial ischemic stroke.

To assess whether acute findings of cerebral arteriopathy, large infarct, and acutely elevated plasma D-dimer levels are independently prognostic of poor long-term neurologic outcome as measured at ≥ 1 year post-event in children with arterial ischemic stroke (AIS). Sixty-one patients with childhood-onset (ie, >28 days of life) AIS were enrolled in a single-institution cohort study at Children's Hospital Colorado between February 2006 and June 2011. Data on demographic and diagnostic characteristics, antithrombotic treatments, and outcomes were systematically collected. Cerebral arteriopathy and D-dimer levels >500 ng/mL (a measure of coagulation activation) were identified acutely in 41% and 31% of the cohort, respectively. Anticoagulation was administered in the acute period post-event in 40% of the children, in the subacute period in 43%, and in the chronic period in 28%. When not receiving anticoagulation, patients were routinely treated with aspirin 2-5 mg/kg once daily for a minimum of 1 year. Death, major bleeding (including intracranial hemorrhage), and recurrent AIS were infrequent. The Pediatric Stroke Outcome Measure at 1 year demonstrated poor outcome in 54% of the children. Acute cerebral arteriopathy and elevated D-dimer level were identified as putative prognostic factors for poor outcome; after adjustment for D-dimer, arteriopathy was an independent prognostic indicator (OR, 19.0; 95% CI, 1.6-229.8; P = .02). Arteriopathy and coagulation activation are highly prevalent in the acute period of childhood AIS. Although recurrent AIS and intracranial hemorrhage were infrequent in our cohort, one-half of children experienced a poor neurologic outcome at 1 year, the risk of which was increased by acute arteriopathy. Substantiation of these findings in multi-institutional cohort studies is warranted, toward risk stratification in childhood-onset AIS.

23260099

Intrahepatic fat is increased in the neonatal offspring of obese women with gestational diabetes.

To assess precision magnetic resonance imaging in the neonate and determine whether there is an early maternal influence on the pattern of neonatal fat deposition in the offspring of mothers with gestational diabetes mellitus (GDM) and obesity compared with the offspring of normal-weight women. A total of 25 neonates born to normal weight mothers (n = 13) and to obese mothers with GDM (n = 12) underwent magnetic resonance imaging for the measurement of subcutaneous and intra-abdominal fat and magnetic resonance spectroscopy for the measurement of intrahepatocellular lipid (IHCL) fat at 1-3 weeks of age. Infants born to obese/GDM mothers had a mean 68% increase in IHCL compared with infants born to normal-weight mothers. For all infants, IHCL correlated with maternal prepregnancy body mass index but not with subcutaneous adiposity. Deposition of liver fat in the neonate correlates highly with maternal body mass index. This finding may have implications for understanding the developmental origins of childhood nonalcoholic fatty liver disease.

23260101

Intravenous fluids versus gastric-tube feeding in hospitalized infants with viral bronchiolitis: a randomized, prospective pilot study.

The American Academy of Pediatrics recommends intravenous fluids for infants with bronchiolitis who are unable to sustain oral feedings. Our randomized, prospective pilot study shows that gastric tube feeding (in 31 infants) is feasible and demonstrated comparable clinical outcomes with intravenous fluids (in 20 infants) among hospitalized infants ≤6 months of age with moderate bronchiolitis.

23260098

A randomized controlled trial to compare heated humidified high-flow nasal cannulae with nasal continuous positive airway pressure postextubation in premature infants.

To determine whether postextubation respiratory support via heated, humidified, high-flow nasal cannulae (HHHFNC) results in a greater proportion of infants younger than 32 weeks' gestation being successfully extubated after a period of endotracheal positive pressure ventilation compared with conventional nasal continuous positive airway pressure (NCPAP). We randomly assigned preterm ventilated infants to Vapotherm HHHFNC or NCPAP after extubation. The primary outcome, extubation failure, was defined by prespecified failure criteria in the 7 days after extubation. A total of 132 ventilated infants younger than 32 weeks' gestation were randomized to receive either HHHFNC (n = 67) or NCPAP (n = 65). Extubation failure occurred in 15 (22%) of the HHHFNC group compared with 22 (34%) of the NCPAP group. There was no difference in the number of infants reintubated in the first week. Treatment with HHHFNC reduced the nasal trauma score 3.1 (SD 7.2) versus NCPAP 11.8 (SD 10.7), P < .001. HHHFNC and NCPAP produced similar rates of extubation failure.

23260097

Higher maternal body mass index is associated with an increased risk for later type 2 diabetes in offspring.

To investigate whether the body mass index (BMI) of a child's mother is associated with an increased future risk of type 2 diabetes, independent of genetic risk or childhood metabolic, behavioral, and environmental factors. The analyses were based on the Cardiovascular Risk in Young Finns Study including 1835 individuals aged 3-18 years at baseline with data on maternal BMI, childhood metabolic factors, as well as 34 newly identified type 2 diabetes susceptibility alleles. These subjects were then followed-up over 21-27 years. Maternal BMI (OR for 1-SD increase 1.54 [95% CI 1.12-2.11], P = .008) and child's systolic blood pressure (1.54 [1.01-2.35], P = .04) were significantly associated with increased odds for later type 2 diabetes, in a multivariable analysis adjusted for age, sex, type 2 diabetes genetic risk score, childhood BMI, insulin, lipids, dietary factors, socioeconomic status, and mother's age, and history of type 2 diabetes. A risk prediction model, which included maternal BMI status outperformed one which utilized only child's BMI data (area under the receiver operating characteristic curve 0.720 vs 0.623, P = .02). The inclusion of genetic risk score and other baseline risk variables did not additionally improve prediction (area under the receiver operating characteristic curve 0.720 vs 0.745, P = .40). Maternal BMI is a useful variable in determining offspring risk of developing type 2 diabetes.

23260096

Using the androgen excess-PCOS society criteria to diagnose polycystic ovary syndrome and the risk of metabolic syndrome in adolescents.

To use the Androgen Excess-PCOS Society (AE-PCOS) criteria in adolescents to diagnose polycystic ovary syndrome (PCOS) and identify the prevalence of metabolic risk factors. Retrospective chart review of adolescents (>2 years postmenarche) presenting at a specialty clinic from 2008 through 2010 with complete evaluation for PCOS and metabolic risk were reviewed. Metabolic risk in adolescents with PCOS was compared with those with ≤ 1 AE-PCOS criteria. Of the 205 adolescents evaluated, 66% were found to have PCOS based on the AE-PCOS criteria. The most common presenting symptom was menstrual irregularity, followed by acne, hirsutism, and weight gain. Adolescents with PCOS had a significantly higher prevalence of obesity, hypertension, and low level of high-density lipoprotein cholesterol. Subjects with PCOS had ≥ 1 metabolic risk factor compared with the subjects without PCOS (63.6% vs 33.3%, P = .002). More adolescents with PCOS had ≥ 2 abnormal metabolic risk factors excluding body mass index compared with those without PCOS (P < .02). The prevalence of metabolic syndrome (≥ 3 risk factors) was 10.8% in adolescents with PCOS compared with 1.7% in those without PCOS (P < .04). Adolescents diagnosed with PCOS based on the AE-PCOS criteria are at a significantly increased risk of ≥ 1 metabolic abnormality. Our data underscore the need to accurately diagnose PCOS in the adolescent population instead of delaying the diagnosis to adulthood. Further, using similar criteria for the diagnosis of PCOS in adolescents (>2 years postmenarche) and adults will be more convenient for the clinician.

23260095

Evaluation of the liver injury unit scoring system to predict survival in a multinational study of pediatric acute liver failure.

To examine the predictive value of the Liver Injury Units (LIU) and admission values (aLIU) of bilirubin and prothrombin time and international normalized ratio scores in a large cohort from the Pediatric Acute Liver Failure (PALF) Study Group, a multinational prospective study. LIU and aLIU scores were calculated for 461 and 579 individuals, respectively, enrolled in the PALF study from 1999 to 2008. Receiver operator characteristic curves were used to evaluate the scores with respect to survival without liver transplantation (LT), death, or LT by 21 days after enrollment. At 21 days, 50.3% of participants were alive without LT, 36.2% underwent LT, and 13.4% died. The c-indices for transplant-free survival were 0.81 based on the LIU score with the international normalized ratio (95% CI, 0.78-0.85) and 0.76 based on the aLIU score (95% CI, 0.72-0.79). The LIU score predicted LT better than it predicted death (c-index for LT 0.84, c-index for death 0.76). Based on data from a large, multicenter cohort of patients with PALF, the LIU score was a better predictor of transplant-free survival than was the aLIU score. The LIU score might be a helpful, dynamic tool to predict clinical outcomes in patients with PALF.

23260093

Defining "poor mobilizer" in pediatric patients who need an autologous peripheral blood progenitor cell transplantation.

The definition of poor mobilizers is not clear in pediatric patients undergoing autologous peripheral blood hematopoietic progenitor cell (HPC) mobilization. Most studies conducted in children define those variables related to the collection of HPC after leukapheresis, but the information regarding exclusively the mobilization process is scarce. In our experience, most children (92.2%) reach the target CD34(+) cell dose for autologous peripheral blood progenitor cell transplantation if CD34(+) cell count was higher than 10/μL. No differences were observed between those with >20 CD34(+) cells/μL and 11-20 CD34(+) cells/μL. In this study, we analyzed the variables that influence CD34(+) cell count; we found that prior use of radiotherapy was the main variable related to poor mobilization. Patients diagnosed with Ewing sarcoma, treated with radiotherapy and mobilized with standard doses of granulocyte colony-stimulating factor (G-CSF) were also at a high risk of mobilization failure. In these patients, we should consider mobilization with high dose G-CSF and be prepared with new mobilization agents to avoid delay on their course of chemotherapy.

23260092

Phenotypic and functional characterization of glucagon-positive cells derived from spontaneous differentiation of D3-mouse embryonic stem cells.

Glucagon expression is being considered as a definitive endoderm marker in protocols aiming to obtain insulin-secreting cells from embryonic stem cells. However, it should be considered that in vivo glucagon is expressed both in definitive endoderm- and neuroectoderm-derived cells. Therefore, the true nature and function of in vitro spontaneously differentiated glucagon-positive cells remains to be established. D3 and R1 mouse embryonic stem cells as well as α-TC1-9 cells were cultured and glucagon expression was determined by real-time PCR and immunocytochemistry. Functional analyses regarding intracellular calcium oscillations were performed to further characterize glucagon(+) cells. Specifically, 5% of D3 and R1 cells expressed preproglucagon, with a small percentage of these (<1%) expressing glucagon-like peptide 1. The constitutive expression of protein convertase 5 supports the expression of both peptides. Glucagon(+) cells co-expressed neurofilament middle and some glucagon-like peptide-1(+) cells, glial fibrillary acidic protein, indicating a neuroectodermic origin. However, few glucagon-like peptide-1(+) cells did not show coexpression with glial fibrillary acidic protein, suggesting a non-neuroectodermic origin for these cells. Finally, glucagon(+) cells did not display Ca(2+) oscillations typical of pancreatic α-cells. These results indicate the possible nondefinitive endodermal origin of glucagon-positive cells spontaneously differentiated from D3 and R1 cell lines, as well as the presence of cells expressing glucagon-like peptide-1 from two different origins.

23260091

Optimization of methodology for production of CD25/CD71 allodepleted donor T cells for clinical use.

Immunotherapy with allodepleted donor T cells improves immunity after T cell-depleted hematopoietic stem cell transplantation. We developed a methodology for selective depletion of alloreactive T cells after activation with host antigen-presenting cells by targeting T cells up-regulating CD25 and CD71. Combined depletion of these cells yields a pool of allodepleted donor T cells with antiviral properties with minimal capacity to cause graft-versus-host disease. Mature dendritic cells were irradiated and used to stimulate donor peripheral blood mononuclear cells for 4 days. The co-culture was stained with anti-CD71-biotin followed by CliniMACS CD25 and Anti-Biotin Reagents (Miltenyi Biotec GmbH; Bergisch Gladbach, Germany) before depletion on the CliniMACS Plus (Miltenyi Biotec GmbH). Residual alloreactivity was tested by flow cytometry, a secondary mixed lymphocyte reaction and limiting dilution analysis, and specific anti-viral immunity with pentamer staining. The large-scale protocol was tested under current good manufacturing practice conditions in five donor-recipient pairs of human leukocyte antigen-matched volunteer donors. We developed a closed-system methodology using cell differentiation bags for cell culture and the COBE2991 Cell Processor (CaridianBCT, Lakewood, CO, USA). We also validated an anti-CD71-biotin generated for ex vivo clinical use. In five large-scale runs, the depleted fraction demonstrated excellent viability (99.9%), minimal residual expression of CD3/CD25 and CD3/CD71 (<0.2%) and passed tests for Mycoplasma, endotoxin, bacterial and fungal sterility. In secondary mixed lymphocyte reaction assays, the median response to host after allodepletion was 0%, whereas responses to third-party peripheral blood mononuclear cells were preserved (median, 105%; range 37%-350%). Limiting dilution analysis assays also demonstrated a reduction in response to host (median, -1.11 log) with preservation of third-party responses, and testing with human leukocyte antigen-restricted pentamers showed that populations of Epstein-Barr virus-specific and cytomegalovirus-specific CD8(+) T cells were retained after depletion. We optimized a protocol for the combined immunomagnetic depletion of alloreactive CD25/CD71 T cells under current good manufacturing practice conditions and tested the efficacy in five donor-recipient pairs.

23260090

Human adipose stromal vascular cell delivery in a fibrin spray.

Adipose tissue represents a practical source of autologous mesenchymal stromal cells (MSCs) and vascular-endothelial progenitor cells, available for regenerative therapy without in vitro expansion. One of the problems confronting the therapeutic application of such cells is how to immobilize them at the wound site. We evaluated in vitro the growth and differentiation of human adipose stromal vascular fraction (SVF) cells after delivery through the use of a fibrin spray system. SVF cells were harvested from four human adult patients undergoing elective abdominoplasty, through the use of the LipiVage system. After collagenase digestion, mesenchymal and endothelial progenitor cells (pericytes, supra-adventitial stromal cells, endothelial progenitors) were quantified by flow cytometry before culture. SVF cells were applied to culture vessels by means of the Tisseel fibrin spray system. SVF cell growth and differentiation were documented by immunofluorescence staining and photomicrography. SVF cells remained viable after application and were expanded up to 3 weeks, when they reached confluence and adipogenic differentiation. Under angiogenic conditions, SVF cells formed endothelial (vWF+, CD31+ and CD34+) tubules surrounded by CD146+ and α-smooth muscle actin+ perivascular/stromal cells. Human adipose tissue is a rich source of autologous stem cells, which are readily available for regenerative applications such as wound healing, without in vitro expansion. Our results indicate that mesenchymal and endothelial progenitor cells, prepared in a closed system from unpassaged lipoaspirate samples, retain their growth and differentiation capacity when applied and immobilized on a substrate using a clinically approved fibrin sealant spray system.

23260089

Assay validation for the assessment of adipogenesis of multipotential stromal cells--a direct comparison of four different methods.

Mesenchymal stromal cells (MSCs) are regenerative and immuno-privileged cells that are used for both tissue regeneration and treatment of severe inflammation-related disease. For quality control of manufactured MSC batches in regard to mature fat cell contamination, a quantitative method for measuring adipogenesis is needed. Four previously proposed methods were validated with the use of bone marrow (BM) MSCs during a 21-day in vitro assay. Oil red staining was scored semiquantitatively; peroxisome proliferator activated receptor-γ and fatty acid binding protein (FABP)4 transcripts were measured by quantitative real-time polymerase chain reaction; FABP4 protein accumulation was evaluated by flow cytometry; and Nile red/4',6-diamidino-2-phenylindole (DAPI) ratios were measured in fluorescent microplate assay. Skin fibroblasts and MSCs from fat pad, cartilage and umbilical cord were used as controls. Oil red staining indicated considerable heterogeneity between BM donors and individual cells within the same culture. FABP4 transcript levels increased 100- to 5000-fold by day 21, with large donor variability observed. Flow cytometry revealed increasing intra-culture heterogeneity over time; more granular cells accumulated more FABP4 protein and Nile red fluorescence compared with less granular cells. Nile red increase in day-21 MSCs was ~5- and 4-fold, measured by flow cytometry or microplate assay, respectively. MSC proliferation/apoptosis was accounted through the use of Nile red/DAPI ratios; adipogenesis levels in day-21 BM MSCs increased ~13-fold, with significant correlations with oil red scoring observed for MSC from other sources. Flow cytometry permits the study of MSC differentiation at the single-cell level and sorting more and less mature cells from mixed cell populations. The microplate assay with the use of the Nile red/DAPI ratio provides rapid quantitative measurements and could be used as a low-cost, high-throughput method to quality-control MSC batches from different tissue sources.

23260088

A comparative analysis of the adipogenic potential in human mesenchymal stromal cells from cord blood and other sources.

Mesenchymal stromal cells (MSCs) from umbilical cord blood (CB) attract attention by significantly impaired or absent adipogenic differentiation compared with MSCs derived from bone marrow (BM) and adipose tissue (AT). The diverging adipogenic propensity between the developmentally younger CB-MSCs and MSCs of the adult AT and BM resembles the age-dependent process in the BM, where adipose tissue increases with advancing age, accompanied by loss of bone stability. Thus, MSCs appeal as an attractive model to study the adipogenic process with respect to tissue sources and developmental ages. We followed the expression of main adipogenic transcription factors, genes and protein markers in CB-, BM- and AT-MSCs under adipogenic induction, after silencing of preadipocyte factor 1 (Pref-1, PREF1) and after incubation with CB-plasma supplemented adipogenic media. An inverse relation in the expression of adipogenesis-associated markers and PREF1 in CB-MSCs suggested an inhibitory role of Pref-1 toward adipogenesis. However, Pref-1 protein was rarely detected in CB-MSCs, and siRNA silencing of Pref-1 failed to induce adipogenic differentiation in CB-MSCs. Thus, the impaired adipogenic differentiation of CB-MSCs in vitro was unrelated to endogenous Pref-1 protein expression. Nevertheless CB-plasma containing Pref-1 protein revealed an anti-adipogenic effect on AT-MSCs. Because Pref-1 is vastly abundant in CB-plasma and confers anti-adipogenic properties, Pref-1 in association with the ontogenic milieu probably induces long-lasting unresponsiveness toward adipogenic stimuli in CB-MSCs.

23260087

Mechanistic action of mesenchymal stem cell injection in the treatment of chemically induced ovarian failure in rabbits.

No curative treatment is known for primary ovarian failure; however, mesenchymal stem cells (MSCs), through self-renewal and regeneration, push the trial to evaluate their role in the treatment of ovarian failure. The aim of this study was to explore the impact of MSCs on cyclophosphamide (CTX)-induced ovarian failure in rabbits and to clarify the mechanism(s) by which MSCs exert their action. Thirty-five adult female rabbits were injected with CTX to induce ovarian failure. Five rabbits were euthanized after the last injection of CTX for histological examination. The others (30 rabbits) were further subdivided into two groups: group 1 (ovarian failure group, 15 rabbits) received no treatment; group 2 (ovarian failure and MSC recipient group, 15 rabbits) received MSCs isolated from extracted bone marrow of male rabbits. A decrease of follicle-stimulating hormone and an increase of estrogen and vascular endothelial growth factor (VEGF) levels in the MSC recipient group versus the ovarian failure group were found. Weak caspase-3 expression and +ve proliferating cell nuclear antigen staining after MSC injection were detected. Cytological and histological examinations showed increased follicle numbers with apparent normal structure of ovarian follicles in the MSC recipient group. Moreover, Y chromosome-containing cells from male donors were present within the ovarian tissues in group 2. The current study suggests that intravenous injection of MSCs into rabbits with chemotherapy-induced ovarian damage improved ovarian function. MSCs accomplish this function by direct differentiation into specific cellular phenotypes and by secretion of VEGF, which influence the regeneration of the ovary.

23260086

Effects of EdU labeling on mesenchymal stem cells.

Thymidine analog 5-ethynyl-2-deoxyuridine (EdU) has recently been used for tracking mesenchymal stem cells (MSCs). In the present study, we tested whether EdU was cytotoxic and whether it interfered with differentiation, cytokine secretion and migration of MSCs. EdU labeling was performed by incubating adipose-derived stem cells (ADSCs) with 10(-8) mol/L of EdU for 48 h. Incorporation of EdU was detected by reaction with azide-conjugated Alexa594. The labeled and unlabeled ADSCs were compared for proliferation and apoptosis as determined by CellTiter and comet assays, respectively. They were also compared for neuron-like and endothelial differentiation as determined by morphology, marker expression and function. Comparison of their secreted cytokine profile was performed by cytokine antibody array. Comparison of their response to homing factor SDF-1 was performed by migration assay. EdU was incorporated into the nucleus in approximately 70% of ADSCs. No significant differences in proliferation and apoptosis rates were observed between EdU-labeled and unlabeled ADSCs. Isobutylmethylxanthine induced both EdU-labeled and unlabeled ADSCs to assume a neuron-like morphology and to express β-III tubulin. Endothelial growth medium-2 (EGM2) induced endothelial differentiation in both EdU-labeled and unlabeled ADSCs, including the ability to uptake low-density lipoprotein and to form capillary-like structures as well as the expression of vWF, eNOS and CD31. EdU-labeled and unlabeled ADSCs exhibited identical secreted cytokine profile and identical migratory response to SDF-1. At the recommended dosage of 10(-8) mol/L, EdU is non-toxic to ADSCs. EdU label did not interfere with differentiation, cytokine secretion or migratory response to SDF-1 by ADSCs.

23260085

Mesenchymal stem cell sheet transplantation combined with locally released simvastatin enhances bone formation in a rat tibia osteotomy model.

Nonunion of fractured bones is a common clinical problem for orthopedic surgeons. This study aimed to investigate the effects of simvastatin locally applied from calcium sulfate (CS) combined with a mesenchymal stem cell (MSC) sheet on fracture healing. In vitro, the proliferation and differentiation of rat bone marrow-derived MSCs stimulated by simvastatin were investigated. In vivo, an osteotomy model was made in rat tibia, and fractured tibias were treated with CS, CS/simvastatin, CS/MSC sheet or simvastatin-loaded CS with MSC or untreated (control). Tibias were harvested at 2 or 8 weeks and underwent real-time quantitative polymerase chain reaction, x-ray, micro-CT and histological analysis. The expression levels of bone morphogenetic protein 2, alkaline phosphatase, osteocalcin, osteoprotegerin and vascular endothelial growth factor of simvastatin-induced MSCs increased with the concentrations of the simvastatin, significantly higher than those in the MSCs group. At 2 weeks, the CS/simvastatin/MSC sheet group showed significantly higher expressions of bone morphogenetic protein 2, alkaline phosphatase, osteocalcin, osteoprotegerin and vascular endothelial growth factor, with more callus formation around the fracture site compared with the other four groups. At 8 weeks, complete bone union was obtained in the CS/simvastatin/MSC sheet group. By contrast, newly regenerated bone tissue partially bridged the gap in the CS/simvastatin group and the CS/MSC sheet group; the control and CS group showed nonunion of the tibia. These results show that both simvastatin and the MSC sheet contributed to the formation of new bone and that the tibia fracture was completely healed by transplantation of the MSC sheet with locally applied simvastatin. Such MSC sheet with locally applied simvastatin might contribute to the treatment of fractures, bone delayed unions or nonunions in clinical practice.

23260083

Tumor stroma engraftment of gene-modified mesenchymal stem cells as anti-tumor therapy against ovarian cancer.

Many ovarian cancers originate from ovarian surface epithelium, where they develop from cysts intermixed with stroma. The stromal layer is critical to the progression and survival of the neoplasm and consequently is recruited into the tumor microenvironment. Using both syngeneic mouse tumors (ID8-R) and human xenograft (OVCAR3, SKOV3) tumor models, we first confirmed that intraperitoneally injected circulating mesenchymal stem cells (MSCs) could target, preferentially engraft and differentiate into α-smooth muscle actin-positive myofibroblasts, suggesting their role as "reactive stroma" in ovarian carcinoma development and confirming their potential as a targeted delivery vehicle for the intratumoral production of interferon-β (IFN-β). Mice with ovarian carcinomas then received weekly intraperitoneal injections of IFN-β expressing MSCs. Intraperitoneal injections of IFN-β expressing MSCs resulted in complete eradication of tumors in 70% of treated OVCAR3 mice (P = 0.004) and an increased survival of treated SKOV3 mice compared with controls (P = 0.01). Similar tumor growth control was observed using murine IFN-β delivered by murine MSCs in ID8-R ovarian carcinoma. As a potential mechanism of tumor killing, MSCs produced IFN-β-induced caspase-dependent tumor cell apoptosis. Our results demonstrate that ovarian carcinoma engrafts MSCs to participate in myofibrovascular networks and that IFN-β produced by MSCs intratumorally modulates tumor kinetics, resulting in prolonged survival.

23260084

Cell therapy with bone marrow stromal cells after intracerebral hemorrhage: impact of platelet-rich plasma scaffolds.

Cell therapy using bone marrow stromal cells (BMSCs) has been considered a promising strategy for neurologic sequelae after intracerebral hemorrhage (ICH). However, after intracerebral administration of BMSCs, most of the cells die, partly because of the absence of extracellular matrix. Intracerebral transplantation of BMSCs, supported in a platelet-rich plasma (PRP) scaffold, optimizes this type of cell therapy. ICH was induced by stereotactic injection of 0.5 IU of collagenase type IV in the striatum of adult Wistar rats (n = 40). Two months later, the rats were subjected to intracerebral administration of 5 × 10(6) allogeneic BMSCs embedded in a PRP scaffold (n = 10), 5 × 10(6) allogeneic BMSCs in saline (n = 10), PRP-derived scaffold only (n = 10) or saline only (n = 10). Functional improvements in each group over the next 6 months were assessed using Rotarod and Video-Tracking-Box tests. Endogenous neurogenesis and survival of transplanted BMSCs were examined at the end of follow-up. Our study demonstrated neurologic improvement after BMSC transplantation and significantly better functional improvement for the group of animals that received BMSCs in the PRP-derived scaffold compared with the group that received BMSCs in saline. Histologic results showed that better functional outcome was associated with strong activation of endogenous neurogenesis. After intracerebral administration of BMSCs, donor cells were integrated in the injured tissue and showed phenotypic expression of glial fibrillary acidic protein and neuronal nucleus. PRP-derived scaffolds increase the viability and biologic activity of BMSCs and optimize functional recovery when this type of cell therapy is applied after ICH.

23260082

Potency assay development for cellular therapy products: an ISCT review of the requirements and experiences in the industry.

The evaluation of potency plays a key role in defining the quality of cellular therapy products (CTPs). Potency can be defined as a quantitative measure of relevant biologic function based on the attributes that are linked to relevant biologic properties. To achieve an adequate assessment of CTP potency, appropriate in vitro or in vivo laboratory assays and properly controlled clinical data need to be created. The primary objective of a potency assay is to provide a mechanism by which the manufacturing process and the final product for batch release are scrutinized for quality, consistency and stability. A potency assay also provides the basis for comparability assessment after process changes, such as scale-up, site transfer and new starting materials (e.g., a new donor). Potency assays should be in place for early clinical development, and validated assays are required for pivotal clinical trials. Potency is based on the individual characteristics of each individual CTP, and the adequacy of potency assays will be evaluated on a case-by-case basis by regulatory agencies. We provide an overview of the expectations and challenges in development of potency assays specific for CTPs; several real-life experiences from the cellular therapy industry are presented as illustrations. The key observation and message is that aggressive early investment in a solid potency evaluation strategy can greatly enhance eventual CTP deployment because it can mitigate the risk of costly product failure in late-stage development.

23260081

The mesenchymal stromal cells dilemma--does a negative phase III trial of random donor mesenchymal stromal cells in steroid-resistant graft-versus-host disease represent a death knell or a bump in the road?

The use of cryopreserved unmatched allogeneic mesenchymal stromal cells (MSCs) for treatment of steroid-resistant graft-versus-host disease has become medical practice in many European jurisdictions. The enthusiasm for use of MSCs in transplantation medicine builds on compelling phase II clinical trial data published by European collaborative groups in the past few years. Notwithstanding, it was reported in 2009 that a large multicenter phase III clinical trial (NCT00366145) conducted in the USA examining the use of an industrial MSC product (Prochymal; Osiris Therapeutics, Inc., Columbia, MD, USA) failed to meet its primary clinical endpoint of achieving a significant increase of complete response of steroid-resistant graft-versus-host disease lasting at least 28 days compared with placebo. Although peer-reviewed publication of the trial and its results are not in public domain at the time of this writing, it is worthwhile to reflect on the apparent discrepancy between the European experience and this industry-sponsored phase III study. This review presents a heuristic failure analysis focusing on the potential variables affecting MSCs and their utility as a cellular pharmaceutical.