The impact of early life factors on cognitive function in old age: The Hordaland Health Study (HUSK).
Previous studies have shown that adverse conditions during fetal and early life are associated with lower performance on neurocognitive tests in childhood, adolescence and adult life. There is, however, a paucity in studies investigating these associations into old age. The aim was to investigate the impact of early life factors on cognitive function in old age by taking advantage of the potential for a linkage between a community survey and historical birth records. A historical cohort study employing a linkage between a community survey of people aged 72-74 years with the participants' birth records (n=346). Early life factors included anthropometric measures taken at birth, birth complications, parental socioeconomic status, and maternal health status. The main outcome was a z-scored composite cognitive score, based on test scores from Kendrick Object Learning Test, Trail Making Test A, a modified version of the Digit Symbol Test, Block Design, a modified version of Mini-Mental State Examination and an abridged version of the Controlled Oral Word Association Test (COWAT). The separate cognitive tests were also individually analysed in relation to measures identified at birth. Higher parental socioeconomic status (SES; based on father's occupation) was associated with a higher value on the composite cognitive score (by 0.25 SD, p=0.0146) and higher Digit Symbol and Trail Making Test A performance. Higher head circumference at birth was associated with higher COWAT and Trail Making Test A performance. Both higher parental SES and head circumference at birth predicted cognitive function in old age independently of each other. There were no other consistent associations. In general we found little evidence for a substantial role of early life factors on late-life cognitive function. However, there was some evidence for an association with parental SES status and head circumference on certain cognitive domains.


A sample-size-optimal Bayesian procedure for sequential pharmaceutical trials.
Consider a pharmaceutical trial where the consequences of different decisions are expressed on a financial scale. The efficacy of the new drug under consideration has a prior distribution obtained from the underlying biological process, animal experiments, clinical experience, and so forth. Berry and Ho (Biometrics 44, 219-227) show how these components are used to establish an optimal (Bayes) sequential testing procedure, assuming a known constant sample size at each decision point. We show in this article how it is also possible to optimize further, with respect to the sample-size rule. This component of the design, which is missing from most sequential procedures, has the potential to yield considerably larger expected net gains (equivalently, considerably smaller Bayes risks).


Overexpression of amyA and glaA substantially increases glucoamylase activity in Aspergillus niger.
The purpose of this study was to obtain an engineered Aspergillus niger strain with high glucoamylase activity by overexpressing the glucoamylase gene glaA and α-amylase gene amyA in A. niger CICC2462. Three recombinant strains containing a single copy of amyA (1A), containing two copies of amyA (2A), and coexpressing amyA and glaA (AG), respectively, were constructed. The transcript levels of amyA in 1A and 2A were increased by 2.95 folds and 3.09 folds, respectively. The levels of amyA and glaA in AG were increased by 1.21 folds and 2.86 folds, but the maximum extracellular glucoamylase activities did not differ significantly. In addition, after 1% casein phosphopeptides (CPPs) was added to the fermentation medium, the maximum extracellular glucoamylase activities for strains 1A, 2A, and AG were 35,200, 37,300, and 40,710 U/ml, respectively, which were significantly higher than that of the parental strain CICC2462 (28,250 U/ml), while CPPs alone had no effect on the parental strain CICC2462. We demonstrate that overexpression of amyA and glaA substantially increases the expression and secretion of glucoamylase in A. niger, and CPPs effectively improves the yield of glucoamylase in recombinant A. niger strains overexpressing amyA and glaA. The newly developed strains and culture methods may have extensive industrial applications.


Methylenedioxymethamphetamine-like discriminative stimulus effects of pyrrolidinyl cathinones in rats.
Synthetic cathinone derivatives are used as alternatives both for stimulant drugs such as cocaine and methamphetamine and for club drugs such as 3,4-methylenedioxymethamphetamine (MDMA), but little is known about their MDMA-like subjective effects. In order to determine their similarity to MDMA, the discriminative stimulus effects of 10 pyrrolidinyl cathinones (α-pyrrolidinopropiophenone, 4'-methyl-α-pyrrolidinopropiophenone (4'-MePPP), α-pyrrolidinobutiophenone, 3',4'-methylenedioxy-α-pyrrolidinobutyrophenone (MD-PBP), α-pyrrolidinovalerophenone, 3,4-methylenedioxy-pyrovalerone (MDPV), α-pyrrolidinopentiothiophenone, napthylpyrovalerone (naphyrone), α-pyrrolidinohexiophenone, and 4'-methyl-α-pyrrolidinohexiophenone (4'-MePHP)) were assessed in Sprague-Dawley rats trained to discriminate 1.5 mg/kg racemic ±-MDMA from vehicle. Compounds with no substitutions on the phenyl ring and the thiophene produced 44-67% MDMA-appropriate responding. In contrast, the substituted pyrrolidinyl cathinones produced a range of MDMA-appropriate responding dependent upon the length of the alpha side chain. 4'-MePPP, with a single carbon on the alpha position, produced 99.8% MDMA-appropriate responding, MD-PBP (two carbons) produced 83%, naphyrone (three carbons) produced 71%, MDPV (three carbons) produced, 66%, and 4'-MePHP (four carbons) produced 47%. Many cathinone compounds have discriminative stimulus effects similar to those of MDMA. However, the pyrrolidine substitution appears to reduce serotonergic effects, with a commensurate decrease in MDMA-like effects. Substitutions on the phenyl ring appear to be able to restore MDMA-like responding, but only in compounds with short alpha side chains. These findings agree with earlier findings of increasing dopaminergic effects and stronger reinforcing effects with increasing side chain. Assessment of more compounds is necessary to establish the replicability/robustness of this phenomenon. These findings may be of use in predicting which compounds will have MDMA/club drug-like effects versus psychostimulant-like effects.


Exposure to postnatal depression predicts elevated cortisol in adolescent offspring.
Animal research shows that early adverse experience results in altered glucocorticoid levels in adulthood, either raised basal levels or accentuated responses to stress. If a similar phenomenon operates in humans, this suggests a biological mechanism whereby early adversity might transmit risk for major depression, glucocorticoid elevations being associated with the development of this disorder. We measured salivary cortisol at 8:00 am and 8:00 pm over 10 days in 13-year-old adolescents who had (n = 48) or had not (n = 39) been exposed to postnatal maternal depression. Maternal postnatal depression was associated with higher, more variable morning cortisol in offspring, a pattern previously found to predict major depression. Early adverse experiences might alter later steroid levels in humans. Because maternal depression confers added risk for depression to children, these alterations might provide a link between early events and later psychopathology.


Re-examining the arterial cord blood gas pH screening criteria in neonatal encephalopathy.
Screening criteria for neonatal encephalopathy remain a complex combination of subjective and objective criteria. We examine the utility of universal cord blood gas testing and mandatory encephalopathy evaluation for infants with pH ≤7.10 on umbilical cord arterial blood gas (cABG) as a single screening measure for timely identification of moderate/severe encephalopathy. Infants born at a single centre between 2008 and 2015, who were ≥36 weeks, had no congenital anomalies and had a cABG pH ≤7.10 were identified for a retrospective cohort study. Maternal/perinatal and patient factors were collected. 27 028 infants were born during the study period; 412 met all inclusion criteria. Of those, 35/85 infants with pH <7.00 and 34/327 infants with pH between 7.00 and 7.10 had moderate/severe encephalopathy. Encephalopathy was identified on the basis of pH and examination alone (no other perinatal criteria present) in 5/35 and 13/34 infants in the two pH groups, respectively.A cABG pH threshold of ≤7.10 was associated with a sensitivity of 74.2% and a specificity of 98.7% for detection of moderate/severe encephalopathy. Based on these data, 25 infants with cABG pH between 7.00 and 7.10 will need to be screened to identify one neonate with moderate/severe encephalopathy, who might have otherwise been missed using conventional screening, a 15% increase in appropriate selection and treatment over current methods. Universal cord blood gas screening with a pH threshold ≤7.10 and mandatory encephalopathy examination results in greater detection of infants with moderate/severe encephalopathy and timely initiation of therapeutic hypothermia.


Improving the Incoherence of a Learned Dictionary via Rank Shrinkage.
This letter considers the problem of dictionary learning for sparse signal representation whose atoms have low mutual coherence. To learn such dictionaries, at each step, we first update the dictionary using the method of optimal directions (MOD) and then apply a dictionary rank shrinkage step to decrease its mutual coherence. In the rank shrinkage step, we first compute a rank 1 decomposition of the column-normalized least squares estimate of the dictionary obtained from the MOD step. We then shrink the rank of this learned dictionary by transforming the problem of reducing the rank to a nonnegative garrotte estimation problem and solving it using a path-wise coordinate descent approach. We establish theoretical results that show that the rank shrinkage step included will reduce the coherence of the dictionary, which is further validated by experimental results. Numerical experiments illustrating the performance of the proposed algorithm in comparison to various other well-known dictionary learning algorithms are also presented.


Interleukin-22 mediates early host defense against attaching and effacing bacterial pathogens.
Infections by attaching and effacing (A/E) bacterial pathogens, such as Escherichia coli O157:H7, pose a serious threat to public health. Using a mouse A/E pathogen, Citrobacter rodentium, we show that interleukin-22 (IL-22) has a crucial role in the early phase of host defense against C. rodentium. Infection of IL-22 knockout mice results in increased intestinal epithelial damage, systemic bacterial burden and mortality. We also find that IL-23 is required for the early induction of IL-22 during C. rodentium infection, and adaptive immunity is not essential for the protective role of IL-22 in this model. Instead, IL-22 is required for the direct induction of the Reg family of antimicrobial proteins, including RegIIIbeta and RegIIIgamma, in colonic epithelial cells. Exogenous mouse or human RegIIIgamma substantially improves survival of IL-22 knockout mice after C. rodentium infection. Together, our data identify a new innate immune function for IL-22 in regulating early defense mechanisms against A/E bacterial pathogens.


Caesarean section is associated with an increased risk of childhood-onset type 1 diabetes mellitus: a meta-analysis of observational studies.
The aim of this study was to investigate the evidence of an increased risk of childhood-onset type 1 diabetes in children born by Caesarean section by systematically reviewing the published literature and performing a meta-analysis with adjustment for recognised confounders. After MEDLINE, Web of Science and EMBASE searches, crude ORs and 95% CIs for type 1 diabetes in children born by Caesarean section were calculated from the data reported in each study. Authors were contacted to facilitate adjustments for potential confounders, either by supplying raw data or calculating adjusted estimates. Meta-analysis techniques were then used to derive combined ORs and to investigate heterogeneity between studies. Twenty studies were identified. Overall, there was a significant increase in the risk of type 1 diabetes in children born by Caesarean section (OR 1.23, 95% CI 1.15-1.32, p < 0.001). There was little evidence of heterogeneity between studies (p = 0.54). Seventeen authors provided raw data or adjusted estimates to facilitate adjustments for potential confounders. In these studies, there was evidence of an increase in diabetes risk with greater birthweight, shorter gestation and greater maternal age. The increased risk of type 1 diabetes after Caesarean section was little altered after adjustment for gestational age, birth weight, maternal age, birth order, breast-feeding and maternal diabetes (adjusted OR 1.19, 95% CI 1.04-1.36, p = 0.01). This analysis demonstrates a 20% increase in the risk of childhood-onset type 1 diabetes after Caesarean section delivery that cannot be explained by known confounders.


Photoreception and vision in the ultraviolet.
Ultraviolet (UV) light occupies the spectral range of wavelengths slightly shorter than those visible to humans. Because of its shorter wavelength, it is more energetic (and potentially more photodamaging) than 'visible light', and it is scattered more efficiently in air and water. Until 1990, only a few animals were recognized as being sensitive to UV light, but we now know that a great diversity, possibly even the majority, of animal species can visually detect and respond to it. Here, we discuss the history of research on biological UV photosensitivity and review current major research trends in this field. Some animals use their UV photoreceptors to control simple, innate behaviors, but most incorporate their UV receptors into their general sense of vision. They not only detect UV light but recognize it as a separate color in light fields, on natural objects or living organisms, or in signals displayed by conspecifics. UV visual pigments are based on opsins, the same family of proteins that are used to detect light in conventional photoreceptors. Despite some interesting exceptions, most animal species have a single photoreceptor class devoted to the UV. The roles of UV in vision are manifold, from guiding navigation and orientation behavior, to detecting food and potential predators, to supporting high-level tasks such as mate assessment and intraspecific communication. Our current understanding of UV vision is restricted almost entirely to two phyla: arthropods and chordates (specifically, vertebrates), so there is much comparative work to be done.


Impactful publishing: the Journal of Neurosurgery and its diamond anniversary (1944-2019).
With this landmark issue of the Journal of Neurosurgery (JNS), we celebrate the 75th anniversary of continuous publication of articles in neurosurgery. It is likely not a coincidence that the diamond anniversary of the JNS coincides precisely with the 150th anniversary of the birth of Harvey Cushing. It is possible that some events in life are inextricably and cosmically tied together, such as the birth of the founding father of our specialty, the society named after him that ultimately became the American Association of Neurological Surgeons (AANS), and the journal of this organization-the JNS.


Risk of recurrence of occupational back pain over three year follow up.
A random sample including 2342 cases representative of all occupational back injuries in Quebec (1981) was followed up prospectively over three years to assess the recurrence rate of back problems (lumbar, thoracic, and cervical). Each medical and accident report was reviewed to obtain the site of symptoms and occupation. Age, sex, industrial sector, and number of episodes of absence from work were abstracted from the computerised Quebec Compensation Board files. The recurrence rate was 20.0% at one year follow up and 36.3% at three years. A multivariate analysis using a Poisson regression, was performed to model the risk of recurrence over time. Men had a higher chance of recurrence (risk ratio = 1.85, 95% CI = 1.50-2.27) but among recurrent cases, the average total number of episodes was comparable between men and women. Age showed a protective effect on the probability of recurrence (10 years: RR = 0.93, 95% CI = 0.88-0.98) due to the lower recurrence rate in the 45-64 year old group (31.8%). Cervical and lumbar symptoms had identical recurrence profiles whereas thoracic symptoms had a significantly lower recurrence rate. Drivers had the highest recurrence rate (42.1%) and nurses had the highest average number of recurrences (2.03) among recurrent cases. Both occupations had statistically significant excesses after controlling for the other variables.


[A hepatitis non-A, non-B-associated substance in the feces--identification and cloning of a partially double-stranded circular DNA].
By means of a radioimmunoassay a substance excreted in feces could be detected in patients with hepatitis non-A,non-B (HNANB). Feces extracts of patients with sporadic and posttransfusion HNANB as well as of healthy persons were precipitated with PEG, digested with RNase and DNase and separated on CsCl. In HNANB-patients a RIA-positive material with a density of 1.3 g/ml CsCl could be detected which contained a partially double-stranded circular DNA. Cloning of this DNA in lambda-phase resulted in DNA of about 5 Kb, which hybridized with feces DNA under stringent conditions. The 5 Kb-DNA were mapped with different restriction enzymes. A 1.5 Kb EcoRi-fragment cross-hybridizes with HBV-DNA. No hybridization and sequence homologies were found with human, viral and procaryotic DNA as well as with plasmid and phage DNA (data base EMBL, Heidelberg). It is assumed that the DNA excreted in feces of HNANB-patients represents a viral genome not detected so far.


Tumour risks and genotype-phenotype correlations associated with germline variants in succinate dehydrogenase subunit genes SDHB, SDHC and SDHD.
Germline pathogenic variants in SDHB/SDHC/SDHD are the most frequent causes of inherited phaeochromocytomas/paragangliomas. Insufficient information regarding penetrance and phenotypic variability hinders optimum management of mutation carriers. We estimate penetrance for symptomatic tumours and elucidate genotype-phenotype correlations in a large cohort of SDHB/SDHC/SDHD mutation carriers. A retrospective survey of 1832 individuals referred for genetic testing due to a personal or family history of phaeochromocytoma/paraganglioma. 876 patients (401 previously reported) had a germline mutation in SDHB/SDHC/SDHD (n=673/43/160). Tumour risks were correlated with in silico structural prediction analyses. Tumour risks analysis provided novel penetrance estimates and genotype-phenotype correlations. In addition to tumour type susceptibility differences for individual genes, we confirmed that the SDHD:p.Pro81Leu mutation has a distinct phenotype and identified increased age-related tumour risks with highly destabilising SDHB missense mutations. By Kaplan-Meier analysis, the penetrance (cumulative risk of clinically apparent tumours) in SDHB and (paternally inherited) SDHD mutation-positive non-probands (n=371/67 with detailed clinical information) by age 60 years was 21.8% (95% CI 15.2% to 27.9%) and 43.2% (95% CI 25.4% to 56.7%), respectively. Risk of malignant disease at age 60 years in non-proband SDHB mutation carriers was 4.2%(95% CI 1.1% to 7.2%). With retrospective cohort analysis to adjust for ascertainment, cumulative tumour risks for SDHB mutation carriers at ages 60 years and 80 years were 23.9% (95% CI 20.9% to 27.4%) and 30.6% (95% CI 26.8% to 34.7%). Overall risks of clinically apparent tumours for SDHB mutation carriers are substantially lower than initially estimated and will improve counselling of affected families. Specific genotype-tumour risk associations provides a basis for novel investigative strategies into succinate dehydrogenase-related mechanisms of tumourigenesis and the development of personalised management for SDHB/SDHC/SDHD mutation carriers.


Dystonia gene in Ashkenazi Jewish population is located on chromosome 9q32-34.
Idiopathic torsion dystonia (ITD) is a neurological disorder characterized by sustained muscle contractions that appear as twisting movements of the limbs, trunk, and/or neck, which can progress to abnormal postures. Most familial forms of ITD follow autosomal dominant transmission with reduced penetrance. The frequency of ITD in the Ashkenazi Jewish population is five to ten times greater than that in other groups. Recently, a gene for ITD (DYT1) in a non-Jewish kindred was located on chromosome 9q32-34, with tight linkage to the gene encoding gelsolin (GSN). In the present study linkage analysis using DNA polymorphisms is used to locate a gene responsible for susceptibility to ITD in 12 Ashkenazi Jewish families. This dystonia gene exhibits close linkage with the gene encoding argininosuccinate synthetase (ASS), and appears by multipoint analysis to lie in the q32-34 region of chromosome 9, a region that also contains the loci for gelsolin and dopamine-beta-hydroxylase. The same gene may be responsible for ITD both in the non-Jewish kindred mentioned above and in the Ashkenazi Jewish families presented here. However, because there is substantial difference between the penetrance of the dominant allele in these two groups, two different mutations may be operating to produce susceptibility to this disease in the two groups.


An ice-templated, pH-tunable self-assembly route to hierarchically porous graphene nanoscroll networks.
Porous graphene nanostructures are of great interest for applications in catalysis and energy storage. However, the fabrication of three-dimensional (3D) macroporous graphene nanostructures with controlled morphology, porosity and surface area still presents significant challenges. Here we introduce an ice-templated self-assembly approach for the integration of two-dimensional graphene nanosheets into hierarchically porous graphene nanoscroll networks, where the morphology of porous structures can be easily controlled by varying the pH conditions during the ice-templated self-assembly process. We show that freeze-casting of reduced graphene oxide (rGO) solution results in the formation of 3D porous graphene microfoam below pH 8 and hierarchically porous graphene nanoscroll networks at pH 10. In addition, we demonstrate that graphene nanoscroll networks show promising electrocatalytic activity for the oxygen reduction reaction (ORR).


Dietary fatty acids and susceptibility to multiple sclerosis.
The gut microbiome as well as dietary habits have recently been established as environmental contributors to the pathogenesis of multiple sclerosis (MS), a T-cell-mediated autoimmune disease of the central nervous system (CNS). To summarize recent findings on the Janus-faced effects of dietary short-chain fatty acids (SCFAs) and long-chain fatty acids (LCFAs) on T-cell immunity with a special focus on the gut and the microbiome as an interface linking diet and T-cell responses during MS. Review article. The autoimmune basis of MS most likely stems from an imbalance between pro-inflammatory T helper cell (Th)1 and Th17 cells and anti-inflammatory or regulatory mechanisms including regulatory T cells (Treg). Hence, the rationale of currently available therapeutic interventions is to either suppress pathogenic Th1/Th17 and/or to foster Treg responses. Dietary fatty acids are often discussed for their detrimental role in MS. However, recent studies investigating saturated fatty acids in animal models of MS revealed harmful as well as beneficial effects depending on their aliphatic chain length. Dietary SCFAs constitute interesting candidates as safe and potent add-on therapy in the immunomodulatory treatment armamentarium for relapsing-remitting MS.


Effect of Sialic Acid on Platelet Cryopreservation.
Sialic acid is a molecule which is responsible for the net negative surface charge of platelets. We investigated the effect of sialic acid on fresh and cryopreserved platelets. Platelet samples were obtained by platelet apheresis from 8 healthy donors. Platelet suspensions with different sialic acid concentrations (0, 1, 2 and 4 mg/mL) were studied for ADP and ristocetin induced platelet aggregation, basal and ADP induced P-selectin and glycoprotein- Ib/IX expression. Then platelet samples were cryopreserved in 5% DMSO with or without 4 mg/mL sialic acid. After thawing, P-selectin expression was compared with the control group. Six samples were also washed after thawing and P-selectin expression was again compared to unwashed samples. Sialic acid suppressed ADP induced platelet aggregation and P-selectin expression in a dose dependent manner. In cryopreserved samples, P-selectin expression of 4 mg/mL sialic acid containing group was found significantly higher than the control group (p< 0.001). In cryopreserved control group, P-selectin expression of thawedwashed group was significantly higher than thawed-unwashed group (p< 0.05). Our results indicate that sialic acid is not a good cryoprotective agent. Washing procedure after thawing to eliminate DMSO causes significant platelet activation.


The Cascading Effects of Multiple Dimensions of Implementation on Program Outcomes: a Test of a Theoretical Model.
This study tests a theoretical cascade model in which multiple dimensions of facilitator delivery predict indicators of participant responsiveness, which in turn lead to improvements in targeted program outcomes. An effectiveness trial of the 10-session New Beginnings Program for divorcing families was implemented in partnership with four county-level family courts. This study included 366 families assigned to the intervention condition who attended at least one session. Independent observers provided ratings of program delivery (i.e., fidelity to the curriculum and process quality). Facilitators reported on parent attendance and parents' competence in home practice of program skills. At pretest and posttest, children reported on parenting and parents reported child mental health. We hypothesized effects of quality on attendance, fidelity and attendance on home practice, and home practice on improvements in parenting and child mental health. Structural Equation Modeling with mediation and moderation analyses were used to test these associations. Results indicated quality was significantly associated with attendance, and attendance moderated the effect of fidelity on home practice. Home practice was a significant mediator of the links between fidelity and improvements in parent-child relationship quality and child externalizing and internalizing problems. Findings provide support for fidelity to the curriculum, process quality, attendance, and home practice as valid predictors of program outcomes for mothers and fathers. Future directions for assessing implementation in community settings are discussed.


Nitric oxide synthase isozymes. Characterization, purification, molecular cloning, and functions.
Three isozymes of nitric oxide (NO) synthase (EC 22.214.171.124) have been identified and the cDNAs for these enzymes isolated. In humans, isozymes I (in neuronal and epithelial cells), II (in cytokine-induced cells), and III (in endothelial cells) are encoded for by three different genes located on chromosomes 12, 17, and 7, respectively. The deduced amino acid sequences of the human isozymes show less than 59% identity. Across species, amino acid sequences for each isoform are well conserved (> 90% for isoforms I and III, > 80% for isoform II). All isoforms use L-arginine and molecular oxygen as substrates and require the cofactors NADPH, 6(R)-5,6,7,8-tetrahydrobiopterin, flavin adenine dinucleotide, and flavin mononucleotide. They all bind calmodulin and contain heme. Isoform I is constitutively present in central and peripheral neuronal cells and certain epithelial cells. Its activity is regulated by Ca2+ and calmodulin. Its functions include long-term regulation of synaptic transmission in the central nervous system, central regulation of blood pressure, smooth muscle relaxation, and vasodilation via peripheral nitrergic nerves. It has also been implicated in neuronal death in cerebrovascular stroke. Expression of isoform II of NO synthase can be induced with lipopolysaccharide and cytokines in a multitude of different cells. Based on sequencing data there is no evidence for more than one inducible isozyme at this time. NO synthase II is not regulated by Ca2+; it produces large amounts of NO that has cytostatic effects on parasitic target cells by inhibiting iron-containing enzymes and causing DNA fragmentation. Induced NO synthase II is involved in the pathophysiology of autoimmune diseases and septic shock. Isoform III of NO synthase has been found mostly in endothelial cells. It is constitutively expressed, but expression can be enhanced, eg, by shear stress. Its activity is regulated by Ca2+ and calmodulin. NO from endothelial cells keeps blood vessels dilated, prevents the adhesion of platelets and white cells, and probably inhibits vascular smooth muscle proliferation.


Genealogical typing of Neisseria meningitidis.
Despite the increasing popularity of multilocus sequence typing (MLST), the most appropriate method for characterizing bacterial variation and facilitating epidemiological investigations remains a matter of debate. Here, we propose that different typing schemes should be compared on the basis of their power to infer clonal relationships and investigate the utility of sequence data for genealogical reconstruction by exploiting new statistical tools and data from 20 housekeeping loci for 93 isolates of the bacterial pathogen Neisseria meningitidis. Our analysis demonstrated that all but one of the hyperinvasive isolates established by multilocus enzyme electrophoresis and MLST were grouped into one of six genealogical lineages, each of which contained substantial variation. Due to the confounding effect of recombination, evolutionary relationships among these lineages remained unclear, even using 20 loci. Analyses of the seven loci in the standard MLST scheme using the same methods reproduced this classification, but were unable to support finer inferences concerning the relationships between the members within each complex.


A single-run, real-time PCR for detection and identification of Borrelia burgdorferi sensu lato species, based on the hbb gene sequence.
Lyme borreliosis is the most important vector-borne disease caused by spirochetes within the Borrelia burgdorferi sensu lato (B. burgdorferi sl) complex. There is strong evidence that different species of this group of genetically diverse spirochetes are involved in distinct clinical manifestations of the disease. In order to differentiate species within this bacterial complex, we developed a real-time-PCR protocol, which targets the hbb gene. We designed a fluorescein-labeled probe specific of a region of this gene harboring a polymorphism linked to species. An internally Red640 labeled primer allowed a fluorescence resonance energy transfer to occur. The sensitivity of this method was in the range of 10 bacteria per assay. After amplification, a melting curve was generated for genotyping. Analysis of these melting curves clearly allowed the distinction between the main European species of B. burgdorferi sl. One hundred seventy tick extracts were analysed by this hbb-based method and in parallel by amplification of the 5S-23S intergenic spacer and RFLP analyses. There was a good correlation between these two methods. We conclude that this hbb-based real-time-PCR is suitable for epidemiological studies on field-collected ticks, although rare mutations in the genomic sequence spanned by the probe could lead to misidentification.


Defining requirements for collagenase cleavage in collagen type III using a bacterial collagen system.
Degradation of fibrillar collagens is important in many physiological and pathological events. These collagens are resistant to most proteases due to the tightly packed triple-helical structure, but are readily cleaved at a specific site by collagenases, selected members of the matrix metalloproteinases (MMPs). To investigate the structural requirements for collagenolysis, varying numbers of GXY triplets from human type III collagen around the collagenase cleavage site were inserted between two triple helix domains of the Scl2 bacterial collagen protein. The original bacterial CL domain was not cleaved by MMP-1 (collagenase 1) or MMP-13 (collagenase 3). The minimum type III sequence necessary for cleavage by the two collagenases was 5 GXY triplets, including 4 residues before and 11 residues after the cleavage site (P4-P11'). Cleavage of these chimeric substrates was not achieved by the catalytic domain of MMP-1 or MMP-13, nor by full-length MMP-3. Kinetic analysis of the chimeras indicated that the rate of cleavage by MMP-1 of the chimera containing six triplets (P7-P11') of collagen III was similar to that of native collagen III. The collagenase-susceptible chimeras were cleaved very slowly by trypsin, a property also seen for native collagen III, supporting a local structural relaxation of the triple helix near the collagenase cleavage site. The recombinant bacterial-human collagen system characterized here is a good model to investigate the specificity and mechanism of action of collagenases.


Effects of calcium administration during post-ischemic reperfusion on myocardial contractility, stiffness, edema, and ultrastructure.
Previous investigators have suggested that calcium may play a role in the pathogenesis of myocardial cell damage following ischemia and reperfusion. Twenty-six in-situ blood perfused isovolumic canine preparations were divided into four groups. Group I dogs were maintained normocalcemic during 45 min of reperfusion following 45 min of hypothermic (27 degrees C) ischemic arrest; Group II dogs received CaCl2 (7 mg/kg) after 15 min of reperfusion; Group III dogs received citrate solution (0.8 ml/kg citrate-phosphate-dextrose [CPD]) after 15 min of reperfusion; Group IV dogs received 7 mg/kg of CaCl2 at 5 min after receiving the same citrate dose as Group III after 15 min of reperfusion. In Group II hearts, calcium improved the left ventricular contractility (P < 0.05 vs Group I) without causing additional cellular or subcellular injury. Calcium also appeared to increase myocardial stiffness (alpha(n)) compared to Group I hearts (P < 0.01). In Group III hearts, citrate reduced contractility (P < 0.01 vs Group I) and increased myocardial edema (P < 0.005 vs Group I) without any apparent improvement in cellular or subcellular preservation. In Group IV hearts, calcium reversed the depression of contractility caused by citrate, resulted in no additional morphologic injury, increased myocardial stiffness compared to Group I or Group III (P < 0.005), and minimized myocardial edema (P < 0.005 vs Group I or III). These results suggest that calcium administered after 15 min of reperfusion improves the depression of contractility that follows hypothermic ischemic arrest without causing additional myocardial damage.


The baric probe: a novel long-term implantable intracranial pressure monitor with ultrasound-based interrogation.
Prompt diagnosis of shunt malfunction is critical in preventing neurological morbidity and death in individuals with hydrocephalus; however, diagnostic methods for this condition remain limited. For several decades, investigators have sought a long-term, implantable intracranial pressure (ICP) monitor to assist in the diagnosis of shunt malfunction, but efforts have been impeded by device complexity, marked measurement drift, and limited instrumentation lifespan. In the current report, the authors introduce an entirely novel, simple, compressible gas design that addresses each of these problems. The device described herein, termed the "baric probe," consists of a subdural fluid bladder and multichannel indicator that monitors the position of an air-fluid interface (AFI). A handheld ultrasound probe is used to interrogate the baric probe in vivo, permitting noninvasive ICP determination. To assess the function of device prototypes, ex vivo experiments were conducted using a water column, and short- and long-term in vivo experiments were performed using a porcine model with concurrent measurements of ICP via a fiberoptic monitor. Following a toe region of approximately 2 cm H(2)O, the baric probe's AFI demonstrated a predictable linear relationship to ICP in both ex vivo and in vivo models. After a 2-week implantation of the device, this linear relationship remained robust and reproducible. Further, changes in ICP were observed with the baric probe, on average, 3 seconds in advance of the fiberoptic ICP monitor reading. The authors demonstrate "proof-of-concept" and feasibility for the baric probe, a long-term implantable ICP monitor designed to facilitate the prompt and accurate diagnosis of shunt malfunction. The baric probe showed a consistent linear relationship between ICP and the device's AFI in ex vivo and short- and long-term in vivo models. With a low per-unit cost, a reduced need for radiography or CT, and an indicator that can be read with a handheld ultrasound probe that interfaces with any smart phone, the baric probe promises to simplify the care of patients with shunt-treated hydrocephalus throughout both the developed and the developing world.


The association between socioeconomic development at the town level and the distribution of dental caries in Brazilian children.
To investigate the association between dental caries among children in the state of São Paulo, Brazil, and town-level indices of socioeconomic development. We examined 15 385 oral-examination records from children aged 5 or 6 years old from 129 towns and cities in the state of São Paulo. We studied two outcomes: (1) the mean number of decayed, missing, and filled deciduous teeth (dmft index) and (2) the care index, which is the proportion of decayed teeth that have already been filled. The explanatory variables were the child development index, human development index, illiteracy rate among subjects older than 20 years, household income, Gini coefficient, insufficient income, fluoridated water supply, number of dentists per 10 000 inhabitants, number of dentists in the public service per 10 000 inhabitants, and number of weekly hours of dentist work in the public service per 10 000 inhabitants. Multiple linear regression models were fitted to the two outcome variables (dmft index and care index). The multiple linear regression analysis showed that a higher dmft index was associated with a low child development index, a high illiteracy rate, and an unfluoridated water supply. The child development index was significantly associated with the care index, and the number of dentists in the public service per 10 000 inhabitants showed borderline statistical significance. Our results indicate that town-level indices of socioeconomic status are significantly correlated with caries indices. Our results also emphasize the beneficial effect that fluoridating water has on reducing the prevalence of dental caries and the fact that strategies for treating and preventing oral diseases should be emphasized within the context of overall health promotion for children.


[Idiopathic environmental intolerance: 2 disabling entities to recognize].
Idiopathic environmental intolerance is characterized by a variety of non-specific symptoms involving several organs within the same individual, and attributed to the exposure to chemical odors (multiple chemical sensitivities) or to the exposure to electromagnetic fields (electromagnetic hypersensitivity). Symptoms occur following an exposure to agents generally regarded as harmless due to the low levels of exposure, and they do not answer to any definition of organic diseases. The lack of established etiology renders treatment difficult. It is important for practitioner to recognize such disorders and assess the social and professional impact so as to improve patients' quality of life.


Dynamic nature of the proximal AZFc region of the human Y chromosome: multiple independent deletion and duplication events revealed by microsatellite analysis.
The human Y chromosome shows frequent structural variants, some of which are selectively neutral, while others cause impaired fertility due to the loss of spermatogenic genes. The large-scale use of multiple Y-chromosomal microsatellites in forensic and population genetic studies can reveal such variants, through the absence or duplication of specific markers in haplotypes. We describe Y chromosomes in apparently normal males carrying null and duplicated alleles at the microsatellite DYS448, which lies in the proximal part of the azoospermia factor c (AZFc) region, important in spermatogenesis, and made up of "ampliconic" repeats that act as substrates for nonallelic homologous recombination (NAHR). Physical mapping in 26 DYS448 deletion chromosomes reveals that only three cases belong to a previously described class, representing independent occurrences of an approximately 1.5-Mb deletion mediated by recombination between the b1 and b3 repeat units. The remainder belong to five novel classes; none appears to be mediated through homologous recombination, and all remove some genes, but are likely to be compatible with normal fertility. A combination of deletion analysis with binary-marker and microsatellite haplotyping shows that the 26 deletions represent nine independent events. Nine DYS448 duplication chromosomes can be explained by four independent events. Some lineages have risen to high frequency in particular populations, in particular a deletion within haplogroup (hg) C(*)(xC3a,C3c) found in 18 Asian males. The nonrandom phylogenetic distribution of duplication and deletion events suggests possible structural predisposition to such mutations in hgs C and G.


In situ crystal growth of gold nanocrystals on upconversion nanoparticles for synergistic chemo-photothermal therapy.
A multifunctional cancer therapy nanocomposite was proposed and synthesized by linking the pH-responsive SH-PEG-DOX prodrug onto gold nanocrystals that were grown in situ on the surface of upconversion nanoparticles (UCNPs). In the structure of the SH-PEG-DOX prodrug, a hydrazone bond was utilized for subsequent pH-responsive drug release in the intracellular acidic microenvironment of cancer cells. This innovative assembly method is facile and mild, and can be used to obtain nanocomposites of UCNPs and gold, which show excellent photostability and biocompatibility. The final UCNPs@Au-DOX nanocomposites offer efficient treatment effects in vitro under irradiation with an 808 nm laser due to the synergistic effect of chemotherapy and photothermal therapy. In addition, the UCNPs@Au-DOX nanocomposites show excellent intracellular locating ability via upconversion luminescence (UCL) imaging with Er3+ ions and magnetic resonance imaging (MRI) with Gd3+ ions, indicating that they have potential as a visual tracking agent in cancer treatment. Therefore, the presented bioimaging-guided multifunctional synergistic therapy nanocomposites are promising tools for imaging-guided cancer therapy.


Comparative analysis of algorithms for next-generation sequencing read alignment.
The advent of next-generation sequencing (NGS) techniques presents many novel opportunities for many applications in life sciences. The vast number of short reads produced by these techniques, however, pose significant computational challenges. The first step in many types of genomic analysis is the mapping of short reads to a reference genome, and several groups have developed dedicated algorithms and software packages to perform this function. As the developers of these packages optimize their algorithms with respect to various considerations, the relative merits of different software packages remain unclear. However, for scientists who generate and use NGS data for their specific research projects, an important consideration is choosing the software that is most suitable for their application. With a view to comparing existing short read alignment software, we develop a simulation and evaluation suite, Seal, which simulates NGS runs for different configurations of various factors, including sequencing error, indels and coverage. We also develop criteria to compare the performances of software with disparate output structure (e.g. some packages return a single alignment while some return multiple possible alignments). Using these criteria, we comprehensively evaluate the performances of Bowtie, BWA, mr- and mrsFAST, Novoalign, SHRiMP and SOAPv2, with regard to accuracy and runtime. We expect that the results presented here will be useful to investigators in choosing the alignment software that is most suitable for their specific research aims. Our results also provide insights into the factors that should be considered to use alignment results effectively. Seal can also be used to evaluate the performance of algorithms that use deep sequencing data for various purposes (e.g. identification of genomic variants). Seal is available as open source at http://compbio.case.edu/seal/. email@example.com Supplementary data are available at Bioinformatics online.


Effect of high saturated free fatty acids feeding on progression of renal failure in rat model of experimental nephrotoxicity.
The current study evaluates the impact of high saturated fat feeding in rat model of experimental nephrotoxicity induced by gentamicin. Sprague-Dawley rats weighing 200 g were randomized into four groups; the first one received the standard rodents chow for 8 weeks and was treated as control, the second group (HFD)received an experimental high fat diet rich in palm kernel oil (40% of Calories as fat) for the same period. The third group (HFDG) was given 80 mg/kg (body weight)/day gentamicin sulphate intraperitoneally during the last 24 days of the feeding period while the fourth group was given gentamicin as above along with the standard rodents chow. Renal function was assessed through measuring serum creatinine, creatinine clearance and absolute and fractional excretion of both sodium and potassium. At the end, rats underwent a surgical procedure for blood pressure measurement. Renal function study showed a stronger nephrotoxicity for HFDG group. Hypertension was observed in HFD group while the pressure declined after gentamicin co-administration. Overall, changing the feeding behavior toward using more SAFFAs for rats injected with gentamicin promotes the progression of renal failure.


Emergency Department Visits for Self-Inflicted Injuries in Adolescents.
To describe emergency department (ED) visits for self-inflicted injury (SII) among adolescents, examine trends in SII mechanisms, and identify factors associated with increased risk. Analyses included patients aged 10 to 18 years from the National Trauma Data Bank, years 2009 to 2012. We used Cochran-Armitage trend tests to examine change over time and generalized linear models to identify risk factors for SII. We examined 286,678 adolescent trauma patients, 3664 (1.3%) of whom sustained an SII. ED visits for SII increased from 2009 to 2012 (1.1% to 1.6%, P for trend ≤ .001), whereas self-inflicted firearm visits decreased (27.3% to 21.9%, P for trend = .02). The most common mechanism in males was firearm (34.4%), and in females, cut/pierce (48.0%). Odds of SII were higher in females (odds ratio [OR] 1.41, 95% confidence interval [CI] 1.13-1.77), older adolescents (OR 2.73, 95% CI 2.38-3.14), adolescents with comorbid conditions (OR 1.64; 95% CI 1.49-1.80), and Asian adolescents (OR 1.67, 95% CI 1.35-2.08) and lower in African American adolescents (OR 0.78, 95% CI 0.70-0.87). Adolescents in the public or self-pay insurance category had higher odds of SII (OR 1.44, 95% CI 1.27-1.64) than those in the private insurance category (OR 1.15, 95% CI 1.01-1.31). Adolescents with an SII had higher odds of death than those with other injuries (OR 12.9, 95% CI 6.78-24.6). We found a significant increase in the number of SIIs by adolescents that resulted in ED visits from 2009 to 2012. Although SIIs increased, we found a significant decrease in the percentage of adolescents who self-injured with a firearm. SIIs reflect a small percentage of ED visits, but these patients have dramatically higher odds of death.


Deletion of the murein hydrolase CbpD reduces transformation efficiency in Streptococcus thermophilus.
Recently it has been shown that Streptococcus thermophilus is competent for natural genetic transformation. This property is widespread among streptococci and may include all members of the genus. Upon entering the competent state, streptococci start transcribing a number of competence-specific genes whose products are required for binding, uptake and processing of transforming DNA. In addition to the core competence genes, competent streptococci express a number of accessory genes that are dispensable for transformation in the laboratory, but presumably play an important role under natural conditions. In Streptococcus pneumoniae, one of these accessory genes encodes a competence-specific murein hydrolase termed CbpD. Experimental evidence indicates that pneumococcal CbpD is part of a predatory mechanism that lyses noncompetent sister cells or members of closely related species in order to release homologous DNA that can be taken up by the competent attacker cells. Competent S. thermophilus LMG18311 cells produce a CbpD-like protein, Stu0039, which might have the same or a similar function. In the present study we have characterized this protein and shown that it is a murein hydrolase with a novel type of cell surface-binding domain. Furthermore, we show that Stu0039 is rapidly inactivated by H(2)O(2) produced during aerobic growth of S. thermophilus. We propose that this inactivation mechanism has evolved for self-protection purposes to prevent extensive autolysis in a competent population. Interestingly, in contrast to pneumococcal CbpD, which does not affect the transformation properties of the producer strain, deletion of Stu0039 reduces the transformability of S. thermophilus.


Obsessive and compulsive symptoms in prediagnosed Huntington's disease.
Obsessive and compulsive symptoms (OCS) are more prevalent in patients with diagnosed Huntington's disease (HD) than in the general population. Although psychiatric symptoms have been reported in individuals with the HD gene expansion prior to clinical diagnosis (pre-HD), little is known about OCS in this phase of disease. The goal of this study was to assess OCS in 300 pre-HD individuals and 108 non-gene-expanded controls from the Neurobiological Predictors of Huntington's Disease (PREDICT-HD) study (enrolled between November 2002 and April 2007) using a multidimensional, self-report measure of OCS, the Schedule of Compulsions, Obsessions, and Pathologic Impulses (SCOPI). Additionally, pre-HD individuals were classified into 3 prognostic groups on the basis of age and CAG repeat length as "near-to-onset" (< 9 estimated years to onset), "mid-to-onset" (9-15 years to onset), and "far-to-onset" (> 15 years to onset). We compared the 3 pre-HD groups to the controls on SCOPI total score and 5 subscales (checking, cleanliness, compulsive rituals, hoarding, and pathologic impulses), controlling for age and gender. All models showed a significant (p < .05) group effect except for hoarding, with an inverted-U pattern of increasing symptoms: controls < far-to-onset < mid-to-onset, with the near-to-onset group being similar to controls. Although the mid-to-onset group showed the most pathology, mean scores were below those of patients with diagnosed obsessive-compulsive disorder. SCOPI items that separated pre-HD individuals from controls were focused on perceived cognitive errors and obsessive worrying. Subclinical OCS were present in pre-HD participants compared to controls. The OCS phenotype in pre-HD may present with obsessive worrying and checking related to cognitive errors and may be a useful target for clinical screening as it could contribute to functional status.


Investigating the dynamic nature of the interactions between nuclear proteins and histones upon DNA damage using an immobilized peptide chemical proteomics approach.
As a result of the complexity and dynamic range of the cellular proteome, including mutual interactions and interactions with other molecules, focused proteomic approaches are important to study subsets of physiologically important proteins. In one such approach, a small molecule or part of a protein is immobilized on a solid phase and used as bait to fish out interacting proteins from complex mixtures such as cellular lysates. Here, such a chemical proteomics experiment is presented to explore the range of proteins that interact with the N-terminal tail of core histones. Therefore, a core histone consensus N-terminal tail (NTT) peptide was synthesized and immobilized on agarose. Interactions between histone NTTs and proteins are extremely important as they regulate chromatin structure, which is important in many DNA-related processes, like transcription and DNA repair. Induction of DNA damage, like DNA double strand breaks, is known to trigger chromatin remodeling events through interactions between histone NTTs and so-called histone chaperones. Therefore, we set out to investigate specific changes in interactions of nuclear proteins before and shortly after DNA double strand break induction. Over 700 proteins were found to bind specifically to the NTT peptide, which makes our study the most comprehensive proteomic survey of the broad spectrum of nuclear proteins interacting with the NTT of core histones in nucleosomes. Apart from a few exceptions, the abundance of the majority of NTT binding proteins was found to be unchanged following DNA damage. However, an in-depth analysis of protein phosphorylation (we detected more than 90 unique sites in about 60 proteins) revealed that the phosphorylation status of several proteins involved in chromatin remodeling changes upon DNA damage. We observed that in these differentially phosphorylated chaperones are part of closely interacting protein complexes involved in regulatory mechanisms at the crossroads of nucleosome assembly, DNA replication, transcription, and the early onset of DNA damage repair.


Radium dial watches, a potentially hazardous legacy?
This study re-examines the risk to health from radium ((226)Ra) dial watches. Ambient dose equivalent rates have been measured for fifteen pocket watches giving results of up to 30 μSv h(-1) at a distance of 2 cm taken with a series 1000 mini-rad from the front face (arithmetic mean ambient dose equivalent for pocket watches being 13.2 μSv h(-1)). A pocket compass gave rise to a similar ambient dose equivalent rate, of 20 μSv h(-1), to the pocket watches, with its cover open. Eighteen wristwatches have also been assessed, but their dose rates are generally much lower (the arithmetic mean being 3.0 μSv h(-1)), although the highest ambient dose equivalent rate noted was 20 μSv h(-1). A phantom experiment using a TLD suggested an effective dose equivalent of 2.2 mSv/y from a 1 μCi (37 kBq) radium dial worn for 16 h/day throughout the year (dose rate 0.375 μSv h(-1)). For this condition we estimated maximum skin dose for our pocket watches as 16 mSv per year, with effective doses of 5.1 mSv and 1.169 mSv when worn in vest and trouser pockets respectively. This assumes exposure from the back of the watch which is generally around 60-67% of that from the front. The maximum skin dose from a wristwatch was 14 mSv, with 4.2 mSv effective dose in vest pocket. Radium ((226)Ra) decays to the radioactive gas radon ((222)Rn), and atmospheric radon concentration measurements taken around a pocket watch in a small sealed glass sphere recorded 18,728 B qm(-3). All watches were placed in a room with a RAD7 real-time radon detector. Radon concentration average was 259±9 Bq m(-3) over 16 h, compared to background average over 24h of 1.02 Bq m(-3). Over 6 weeks highs of the order of 2000 Bq m(-3) were routinely recorded when the heating/ventilation system in the room was operating at reduced rates, peaking at over 3000 Bq m(-3) on several occasions. Estimates of the activity of (226)Ra in the watches ranged from 0.063 to 1.063 μCi (2.31 to 39.31 kBq) for pocket watches and from 0.013 to 0.875 μCi (0.46 to 32.38 kBq) for wrist watches. The risk from old watches containing radium appears to have been largely forgotten today. This paper indicates a health risk, particular to collectors, but with knowledge and appropriate precautions the potential risks can be reduced.


Constructing scientific authorities: issue framing of chlorinated disinfection byproducts in public health.
The practice of chlorine disinfection of drinking water to reduce microbial risks provides substantial benefits to public health. However, increasing concern around potential risks of cancer associated with exposure to chlorinated disinfection byproducts confuses this issue. This article examines the science agenda regarding chlorinated disinfection byproducts (CDBP) and cancer in Canada and the United States, focusing on the social construction of scientific knowledge claims and evidence. Data for this analysis were obtained from published documents as well as from in-depth interviews with epidemiologists and toxicologists centrally involved with the issue in both countries. Results of the analysis suggest that toxicological scientists want to close the door on the "chloroform issue" due to increasing evidence that chloroform is safe at low doses, because epidemiological scientists can no longer move forward the cancer science until significant improvements can be made in assessing human exposures, and because the scientific foci of research on DBP have shifted accordingly. Further, a distinction emerges in terms of how scientific uncertainties are interpreted when they cross-cut disciplines in the context of human health risk assessment. We suggest this tension reflects a balance of how uncertainty and authorities are managed in a mandated science-policy domain. Sufficient evidence was provided to keep the DBP issue on the regulatory agenda and to generate additional research, yet authorities and concomitant interpretations of uncertainty were contested. Such science generation and contestation inevitably influences complex risk assessment processes with respect to what water-related health risks are addressed and how.


Adjacent segment degeneration after lumbar spinal fusion compared with motion-preservation procedures: a meta-analysis.
This meta-analysis aimed to evaluate the efficacy of motion-preservation procedures to prevent the adjacent segment degeneration (ASDeg) or adjacent segment disease (ASDis) compared with fusion in lumbar spine. PubMed, Embase and the Cochrane Library were comprehensively searched and a meta-analysis was performed of all randomized controlled trials and well designed prospective or retrospective comparative cohort studies assessing the lumbar fusion and motion-preservation procedures. We compared the ASDeg and ASDis rate, reoperation rate, operation time, blood loss, length of hospital stay, visual analogue scale (VAS) and oswestry disability index (ODI) improvement of the two procedures. A total of 15 studies consisting of 1474 patients were included in this study. The meta-analysis indicated that the prevalence of ASDeg, ASDis and reoperation rate on the adjacent level were lower in motion-preservation procedures group than in the fusion group (P = 0.001; P = 0.0004; P < 0.0001). Moreover, shorter length of hospital stay was found in motion-preservation procedures group (P < 0.0001). No difference was found in terms of operation time (P = 0.57), blood loss (P = 0.27), VAS (P = 0.76) and ODI improvement (P = 0.71) between the two groups. The present evidences indicated that the motion-preservation procedures had an advantage on reducing the prevalence of ASDeg, ASDis and the reoperation rate due to the adjacent segment degeneration compared with the lumbar fusion. And the clinical outcomes of the two procedures are similar.


Multiple functions of cation-chloride cotransporters in the fish retina.
A GABA- or glycine-induced increase in Cl(-) permeability can produce either a depolarization or hyperpolarization, depending on the Cl(-) equilibrium potential. It has been shown that retinal neurons express the chloride cotransporters, Na-K-2Cl (NKCC) and K-Cl (KCC), the primary molecular mechanisms that control the intracellular Cl(-) concentration. We thus studied (1) the localization of these cotransporters in the fish retina, and (2) how suppression of cotransporter activity in the fish retina affects function. Specific antibodies against NKCC and KCC2 revealed that both cotransporters were expressed in the outer and inner plexiform layers, and colocalized in many putative amacrine cells and in cells of the ganglion cell layer. However, the somata of putative horizontal cells displayed only NKCC immunoreactivity and many bipolar cells were only immunopositive for KCC2. In the outer retina, application of bumetanide, a specific inhibitor of NKCC activity, (1) increased the steady-state extracellular concentration of K+ ([K+](o)) and enhanced the light-induced decrease in the [K+](o), (2) increased the sPIII photoreceptor-dependent component of the ERG, and (3) reduced the extracellular space volume. In contrast, in the outer retina, application of furosemide, a specific inhibitor of KCC activity, decreased sPIII and the light-induced reduction in [K+](o), but had little effect on steady-state [K+](o). In the inner retina, bumetanide increased the sustained component of the light-induced increase in [K+](o). These findings thus indicate that NKCC and KCC2 control the [K+](o) and extracellular space volume in the retina in addition to regulating GABA- and glycine-mediated synaptic transmission. In addition, the anatomical and electrophysiological results together suggest that all of the major neuronal types in the fish retina are influenced by chloride cotransporter activity.


Surface fragmented QRS in a patient with hypertrophic cardiomyopathy and malignant arrhythmias: Is there an association?
An 18- year old woman with hypertrophic cardiomyopathy, aborted sudden cardiac death and implanted with an implantable cardioverter defibrillator (ICD), developed progressive fragmentation of her surface 12-lead electrocardiogram (ECG). During the follow-up, she presented with multiple appropriate ICD discharges. Here, we discuss the possible association between surface fragmented ECG and the risk of ventricular arrhythmias in patients with hypertrophic cardiomyopathy.


Economic Burden of Mental Illnesses in Pakistan.
The economic consequences of mental illnesses are much more than health consequences. In Low and Middle Income Countries (LMIC) the economic impact of mental illnesses is rarely analyzed. This paper attempts to fill the gap in research on economics of mental health in LMIC. We provide economic burden of mental illness in Pakistan that can serve as an argument for reorienting health policy, resource allocation and priority settings. To estimate economic burden of mental illnesses in Pakistan. The study used prevalence based cost of illnesses approach using bottom-up costing methodology. We used Aga Khan University Hospital, Psychiatry department data set (N = 1882) on admission and ambulatory care for the year 2005-06. Healthcare cost data was obtained from finance department of the hospital. Productivity losses, caregiver and travel cost were estimated using socio-economic features of patients in the data set and data of national household survey. We used stratified random sampling and methods of ordinary least square multiple linear regressions to estimate cost on medicines for ambulatory care. All estimates of cost are based on 1000 bootstrap samples by ICD-10 disease classification. Prevalence data on mental illnesses from Pakistan and regional countries was used to estimate economic burden. The economic burden of mental illnesses in Pakistan was Pakistan Rupees (PKR) 250,483 million (USD 4264.27 million) in 2006. Medical care costs and productivity losses contributed 37% and 58.97% of the economic burden respectively. Tertiary care admissions costs were 70% of total medical care costs. The average length of stay (LOS) for admissions care was around 8 days. Daily average medical care cost of admitted patients was PKR 3273 (USD 55.72). For ambulatory care, on average a patient visited the clinic twice a year. The estimated average yearly cost for all mental illnesses was PKR 81,922 (USD 1394.65) and PKR 19,592 (USD 333.54) for admissions and ambulatory care respectively. In the sensitivity analysis productivity losses showed high variability (from USD 1022.17 million to USD 4007.01 million). Assuming a gate keeping role of primary healthcare (PHC) demonstrated a saving of USD 1577.19 million in total economic burden. This study set out to generate evidence using a low cost innovative approach relevant to many LMICs. In Pakistan, like many LMICs, patients access tertiary care directly, even for illness that can be efficiently managed at PHC level. In economic terms the non-medical consequences of mental illnesses are far greater than medical consequences. Based on these finding we recommend, firstly, that mental illnesses should be prioritized equally as other illnesses in health policy and secondly there needs to be integration of mental health in primary health care in Pakistan.


The epidemiology of vertebral fractures. European Vertebral Osteoporosis Study Group.
Vertebral fractures are recognised as a hallmark of osteoporosis, yet little is known of their epidemiology. This deficiency limits accurate characterisation of the public health importance of osteoporosis. Assessment of the impact of vertebral fractures has been hampered by the absence of formal criteria for identifying fractures on a thoracolumbar radiograph. Initial methods relying upon subjective radiological assessments have given way to morphometric measurements of vertebral heights, with deformities defined according to various algorithms. These methods have been used in a series of studies performed in Rochester, MN, to determine the incidence, outcome, and time trends of vertebral deformities. The results suggest a prevalence rate of vertebral deformity of 25.3 per 100 Rochester women aged 50 years and over (95% CI, 22.3-28.2), with an estimated incidence of 17.8 per 1,000 person-years. The incidence of clinically diagnosed vertebral fractures among women in the same population was 5.3 per 1,000 person-years, suggesting that around 30% of such deformities in women receive clinical attention. Morphometric measurement on the radiographs of women with clinically diagnosed fractures revealed that 80% had grade 2 ( > 4 SD) deformities. Comparable data on the occurrence and health impact of vertebral deformities throughout Europe are urgently required. The European Vertebral Osteoporosis Study (EVOS) is a multicentre epidemiological study that aims to address this issue. It is designed as a radiographic prevalence study in 34 European centres.(ABSTRACT TRUNCATED AT 250 WORDS)


Novel chemiluminescent immunochromatographic assay using a dual-readout signal probe for multiplexed detection of pesticide residues.
A novel immunochromatographic assay (ICA) using a dual-readout signal probe was developed for multiplexed detection of pesticide residues by adopting methyl parathion and fenpropathrin as model analytes. Luminol-reduced Au nanoparticles (LRAuNPs) were synthesized and utilized in the proposed ICA platform as a colorimetric/chemiluminescent (CL) dual-readout probe. The methyl parathion antibody and fenpropathrin antibody were tagged with the prepared LRAuNPs to conduct spatially-resolved multiplexed detection. After the occurrence of two immunoreactions on the test strip, the probes were captured by the immobilized antigens on the two test zones. The red color resulting from the accumulation of captured LRAuNPs was adopted as the visual and semi-quantitative readout. For the sensitive quantitative detection of the analytes, the CL signals caused by the luminophore in the LRAuNPs were collected after triggering the luminol-H2O2 CL reaction. Under the optimal conditions, the detection limits for methyl parathion and fenpropathrin were 0.17 ng mL-1 and 0.10 ng mL-1 (S/N = 3), respectively. The whole procedure for ICA was completed within 15 min. The present ICA protocol was successfully applied for detection of pesticide residues in spiked traditional Chinese medicine samples. This dual-readout ICA platform showed merits such as low cost, time efficiency, easy operation and high sensitivity. Its application potential has been demonstrated in the rapid screening and field detection of multiple pesticide residues.


Inhibition of the anaphase-promoting complex by the Xnf7 ubiquitin ligase.
Degradation of specific protein substrates by the anaphase-promoting complex/cyclosome (APC) is critical for mitotic exit. We have identified the protein Xenopus nuclear factor 7 (Xnf7) as a novel APC inhibitor able to regulate the timing of exit from mitosis. Immunodepletion of Xnf7 from Xenopus laevis egg extracts accelerated the degradation of APC substrates cyclin B1, cyclin B2, and securin upon release from cytostatic factor arrest, whereas excess Xnf7 inhibited APC activity. Interestingly, Xnf7 exhibited intrinsic ubiquitin ligase activity, and this activity was required for APC inhibition. Unlike other reported APC inhibitors, Xnf7 did not associate with Cdc20, but rather bound directly to core subunits of the APC. Furthermore, Xnf7 was required for spindle assembly checkpoint function in egg extracts. These data suggest that Xnf7 is an APC inhibitor able to link spindle status to the APC through direct association with APC core components.


Temperature titration: a new approach to the thermodynamics of oxygen binding to hemoglobin.
A cell was constructed in order to study hemoglobin's reaction with gaseous ligands. The temperature of the hemoglobin sample is systematically altered within a given temperature range (275-310 degrees K), while the percentage of oxygen in the equilibrating gas is kept constant. The equilibration time of the sample at each temperature step depends on sample concentration, ligand affinity, and absolute temperature; in most cases, the equilibration time is on the order of minutes. The construction of the optical compartment allows the experimenter to vary the optical pathlength using specially designed spacers, thus making it possible to study hemoglobin-ligand interactions over a wide range of protein concentrations (0.1-200 mg/ml). Optical glass is used in the construction of the cuvette in order to optimize its optical stability over a long period of time. At equilibrium the absorption spectrum of the sample is collected and decomposed into the relative contributions of oxy-Hb, deoxy-Hb, and ferric-Hb, thus revealing the fraction of oxyhemoglobin as well as any baseline drifts and protein degradation. Temperature steps of 1 degree K are already sufficient to change the absorption spectra in a significant way. This type of setup is also advantageous in that the experimenter can change the sample at any point (temperature) without having to restart the entire experiment. This makes it possible to study the oxygen binding characteristics of unstable hemoglobins. Analyses of the binding curves obtained with this technique immediately yield the overall oxygen binding constants beta i together with the respective standard enthalpies delta H(i).


Improved chemical and mechanical stability of peptoid nanosheets by photo-crosslinking the hydrophobic core.
Peptoid nanosheets can be broadly functionalized for a variety of applications. However, they are susceptible to degradation when exposed to chemical or mechanical stress. To improve their strength, photolabile monomers were introduced in order to crosslink the nanosheet interior. Photo-crosslinking produced a more robust material that can survive sonication, lyophilization, and other biochemical manipulations.


Fracture resistance of all-ceramic and metal-ceramic inlays.
Metal-ceramic inlay designs were developed to determine if the esthetic qualities of all-ceramic inlays could be duplicated and at the same time improve their strength and stability. The objectives of this study were to: (1) compare the fracture resistance of metal-ceramic inlays with that of all-ceramic inlays; (2) determine the correlation between the degree of preparation taper and fracture resistance; and (3) determine the correlation between marginal gap width and fracture resistance. Inlay preparations were made on 60 Dentoform teeth, with 30 teeth allocated for metal-ceramic inlays and 30 teeth for all-ceramic inlays. Each group was further subdivided into 5-, 10-, and 20-degree taper preparations. Metal-ceramic inlays were fabricated using Goldtech Bio 2000 metal and Ceramco porcelain extending to the margin, while all-ceramic inlays were made from Empress II ceramic. Marginal gap widths were measured at six critical areas after fabrication. The load at failure was measured using an Instron Universal Testing Machine. The mean fracture load for all-ceramic inlays and metal-ceramic inlays at 5, 10, and 20 degrees was 70+/-40 N, 48+/-37 N, 33+/-7 N, and 40+/-23 N, 29+/-22 N, and 14+/-4 N, respectively. The mean gap width was 105 microm and 126 microm for all-ceramic and metal-ceramic inlays, respectively. The mean fracture load for Empress inlays was significantly higher than that for metal-ceramic inlays. Inlays with a 5-degree taper were significantly more fracture resistant than those with a 20-degree taper. There was no relation between marginal gap width and fracture resistance.


Changing patterns of bridging to heart transplantation in children.
Mechanical support as a bridge to cardiac transplantation in children is an accepted treatment. With improved devices and increasing experience, the length of time that children can be supported has increased. Donor organs remain scarce and there is significant associated morbidity. Retrospective review of all children offered mechanical support as a bridge to heart transplant over 10 years in one of the two UK pediatric heart transplant centers. Outcomes during the years 1998 to 2002 were compared with outcomes during the years 2003 to 2007. Forty children in 41 separate patient episodes received mechanical support as a bridge to transplantation or, in 1 case, to recovery. Survival to transplant or recovery was achieved in 29 of 41 (71%); 26 of 40 children (63%) survived to hospital discharge. Devices used were extracorporeal membrane oxygenation (ECMO), the Medos HIAA, the Berlin Heart (from November 2005) and the Levitronix ventricular assist device (VAD) from 2007. All 3 children supported with the Levitronix survived to transplant (median duration of support 10 days). Ten of 13 children (77%) supported by the Berlin Heart survived to transplant or recovery (median duration of support 44 days). Four of 7 (57%) children supported using the Medos device survived to transplant (median duration of support 7 days). Neurologic events were the most common cause of death in both eras (1998 to 2002 and 2003 to 2008). Waiting times to pediatric cardiac transplant in the UK have increased. The Berlin Heart allows children to be bridged to transplant over long periods. Neurologic morbidity remains as a major concern.


XML-based approaches for the integration of heterogeneous bio-molecular data.
The today's public database infrastructure spans a very large collection of heterogeneous biological data, opening new opportunities for molecular biology, bio-medical and bioinformatics research, but raising also new problems for their integration and computational processing. In this paper we survey the most interesting and novel approaches for the representation, integration and management of different kinds of biological data by exploiting XML and the related recommendations and approaches. Moreover, we present new and interesting cutting edge approaches for the appropriate management of heterogeneous biological data represented through XML. XML has succeeded in the integration of heterogeneous biomolecular information, and has established itself as the syntactic glue for biological data sources. Nevertheless, a large variety of XML-based data formats have been proposed, thus resulting in a difficult effective integration of bioinformatics data schemes. The adoption of a few semantic-rich standard formats is urgent to achieve a seamless integration of the current biological resources.


CREB1 and Smad3 mediate TGF‑β3‑induced Smad7 expression in rat hepatic stellate cells.
Transforming growth factor (TGF)‑β3 has previously been reported to antagonize hepatic fibrosis in vivo and in vitro. The present study aimed to investigate the mechanism underlying the involvement of TGF‑β3 in hepatic fibrosis. Short hairpin (sh)RNA‑cAMP-responsive element binding protein (CREB) 1 and small interfering (si)RNA‑Smad3 were utilized to silence the expression of CREB1 and Smad3 in hepatic stellate cells (HSCs), whereas the vector pRSV‑CREB1 was used to induce CREB1 overexpression in HSCs. Cells were treated with or without exogenous TGF‑β3 or TGF‑β1, and mRNA and protein expression levels were assessed using reverse transcription‑quantitative polymerase chain reaction and western blot analysis. Untreated cells served as the control group. Exogenous TGF‑β3 increased Smad7 mRNA and protein expression levels in rat HSCs, and CREB1 and Smad3 appeared to be implicated in the mechanism of Smad7. CREB1 knockdown inhibited the TGF‑β3‑induced upregulation of Smad7, whereas its overexpression potentiated the Smad7 upregulation in HSCs; conversely, CREB1 manipulations had no effect on Smad7 expression under basal conditions. In addition, TGF‑β3‑induced Smad7 upregulation was blocked when the activity of p38, a kinase upstream of CREB1, was inhibited. Furthermore, silencing Smad3 resulted in decreased Smad7 expression under basal conditions and in TGF‑β3‑stimulated cells. Notably, Smad7 expression appeared to also be induced by exogenous TGF‑β1, independent of CREB1. The present study demonstrated that TGF‑β3 increased Smad7 expression in HSCs, whereas CREB1 and Smad3 appeared to participate in the mechanism of induction. Smad3 is the key regulator whereas CREB‑1 acts as a co‑regulator. These results suggested that this mechanism may underlie the antagonizing effects of TGF‑β3 on hepatic fibrosis.


Factor structure and construct validity of the psychopathic personality inventory in a forensic sample.
A wealth of research has underscored the strong relationship between PCL-R scores and recidivism. However, mounting criticism cites the PCL-R's cumbersome administration procedures and failure to adequately measure core features associated with the construct of psychopathy (Skeem, Polaschek, Patrick, & Lilienfeld, 2011). In light of these concerns, this study examined the PPI and the PPI-R, which were designed to measure core personality features associated with psychopathy (Lilienfeld & Andrews, 1996; Lilienfeld & Widows, 2005). Study one examined the PPI relative to the PCL-R and examined its factor structure. The instruments shared few significant correlations and neither the PCL-R nor the PPI significantly predicted recidivism. Study two examined the PPI-R relative to the PCL-R, the PPI, both history of violence and future criminal activity and measure of related constructs. The PPI-R was significantly correlated with measures of empathy and criminal thinking and the factors were related to a history of violence and predicted future violent criminal behavior.


Study of Compression-Induced Supramolecular Nanostructures of an Imidazole Derivative by Langmuir-Blodgett Technique.
In this communication, we report the design and synthesis as well as the supramolecular assembly behavior of a 2,4,5-triaryl imidazole derivative (compound 1) at the air-water interface and in thin films using Langmuir-Blodgett (LB) technique. The main idea for such a chemical structure is that the long alkyl chain and N-H of the imidazole core may help to form supramolecular architecture through the hydrophobic-hydrophobic interaction and hydrogen bonding, respectively. Accordingly, the interfacial behavior as well as morphology of 1 in thin films were studied through a series of characterization methods such as surface pressure-area (π-A) isotherm, hysteresis analysis, ultraviolet-visible (UV-vis) absorption and steady-state fluorescence spectroscopies, Fourier transform infrared, X-ray diffraction, Brewster angle microscopy (BAM), and atomic force microscopy (AFM) measurements, and so forth. Pressure-area isotherm is an indication toward the formation of supramolecular nanostructures instead of an ideal monolayer at the air-water interface. This has been confirmed by the hysteresis analysis and BAM measurement at the air-water interface. AFM images of 1 in the LB monolayer exhibits the formation of supramolecular nanowires as well as nanorods. By controlling different film-forming parameters, it becomes possible to manipulate these nanostructures. With the passage of time, the nanowires come close to each other and become straight. Similarly, nanorods come close to each other and form bundles of several rods in the LB films. H-bonding, J-aggregation, as well as compression during film formation might play a key role in the formation of such nanostructures. Electrical switching behavior of compound 1 was also observed because of the presence of an electron donor-acceptor system in 1. This type of organic switching behavior may be promising for next-generation organic electronics.


The minimum monitoring signal-to-noise ratio for off-axis signals and its implications for directional hearing aids.
The signal-to-noise ratio (SNR) benefit of hearing aid directional microphones is dependent on the angle of the listener relative to the target, something that can change drastically and dynamically in a typical group conversation. When a new target signal is significantly off-axis, directional microphones lead to slower target orientation, more complex movements, and more reversals. This raises the question of whether there is an optimal design for directional microphones. In principle an ideal microphone would provide the user with sufficient directionality to help with speech understanding, but not attenuate off-axis signals so strongly that orienting to new signals was difficult or impossible. We investigated the latter part of this question. In order to measure the minimal monitoring SNR for reliable orientation to off-axis signals, we measured head-orienting behaviour towards targets of varying SNRs and locations for listeners with mild to moderate bilateral symmetrical hearing loss. Listeners were required to turn and face a female talker in background noise and movements were tracked using a head-mounted crown and infrared system that recorded yaw in a ring of loudspeakers. The target appeared randomly at ± 45, 90 or 135° from the start point. The results showed that as the target SNR decreased from 0 dB to -18 dB, first movement duration and initial misorientation count increased, then fixation error, and finally reversals increased. Increasing the target angle increased movement duration at all SNRs, decreased reversals (above -12 dB target SNR), and had little to no effect on initial misorientations. These results suggest that listeners experience some difficulty orienting towards sources as the target SNR drops below -6 dB, and that if one intends to make a directional microphone that is usable in a moving conversation, then off-axis attenuation should be no more than 12 dB.


Morphological, Pathogenic, and Genetic Comparisons of Colletotrichum graminicola Isolates from Poaceae.
Isolates of Colletotrichum graminicola from annual blue grass and creeping bent grass were investigated for their morphological characteristics, host specificity, and genetic relatedness. One isolate from maize and one from sorghum (C. sublineolum) were included for comparison. Recently isolated cultures of C. graminicola from annual blue grass were readily distinguished from those isolated from creeping bent grass on the basis of pigmentation. Differences in appressoria size and shape were found only between the turf grass isolates and those from maize and sorghum. Spore length varied significantly between host groups. Differences in host range and virulence were also apparent. In general, isolates from creeping bent grass incited disease on both creeping bent grass and annual blue grass, while those from annual blue grass essentially were limited to host. Random amplified polymorphic DNA (RAPD) marker analysis of C. graminicola isolates from turf grass revealed that a high degree of genetic similarity exists among isolates recovered from the same host, but exceptions were found. Therefore, an absolute distinction between isolates recovered from two turf grass hosts could not be made based on RAPD markers.


RNA-induced silencing complex-bound small interfering RNA is a determinant of RNA interference-mediated gene silencing in mice.
Deeper knowledge of pharmacokinetic and pharmacodynamic (PK/PD) concepts for RNA therapeutics is important to streamline the drug development process and for rigorous selection of best performing drug candidates. Here we characterized the PK/PD relationship for small interfering RNAs (siRNAs) targeting luciferase by examining siRNA concentration in plasma and liver, the temporal RNA-induced silencing complex binding profiles, mRNA reduction, and protein inhibition measured by noninvasive bioluminescent imaging. A dose-dependent and time-related decrease in bioluminescence was detected over 25 days after a single treatment of a lipid nanoparticle-formulated siRNA targeting luciferase messenger RNA. A direct relationship was observed between the degree of in vivo mRNA and protein reduction and the Argonaute2 (Ago2)-bound siRNA fraction but not with the total amount of siRNA found in the liver, suggesting that the Ago2-siRNA complex is the key determinant of target inhibition. These observations were confirmed for an additional siRNA that targets endogenously expressed Sjögren syndrome antigen B (Ssb) mRNA, indicating that our observations are not limited to a transgenic mouse system. Our data provide detailed information of the temporal regulation of siRNA liver delivery, Ago2 loading, mRNA reduction, and protein inhibition that are essential for the rapid and cost-effective clinical development of siRNAs therapeutics.


Differences in heritability estimates from multiple-trait and repeated-records models.
Analyses of ovulation rates in consecutive estrous cycles with multiple-trait and repeated-records animal models resulted in different estimates of heritability. The estimate from the repeated-records model was seen to be approximately the product of the average genetic correlation and the average heritability from the multiple-trait procedure. A simple model is used to show algebraically that such a result is expected, particularly if the environmental correlations are small among records of the same animal. Comparison of results of the two types of analyses of 10 replications of 10 combinations of underlying heritabilities and genetic correlations confirms this explanation.


Different measures of speckle and coherence at the output of a multimode optical fiber.
Excitation of a multimode fiber with a focused spatially coherent light of finite bandwidth results in a partially coherent light at the output of the fiber. Here we study the properties of speckle and classical coherence of such light with analytical theory, numerical modeling, and experimentally. Of particular interest is the relationship between measures of coherence and speckle and their dependence on input source bandwidth and fiber length. Speckle contrast is easy to measure experimentally and there exist at least two different methods to generate ensembles of random speckles. We show that speckle contrast evaluated over the ensemble of external diffusers is related to the number of effective modes-one of the characteristics of beam global coherence. The other speckle contrast measure evaluated over the ensemble of random bends and twists of the fiber is related to residual coherence, which is the pedestal on the average modulus of the complex degree of the coherence function on the output endface of the fiber.


Setting-up an in vitro model of rat blood-brain barrier (BBB): a focus on BBB impermeability and receptor-mediated transport.
The blood brain barrier (BBB) specifically regulates molecular and cellular flux between the blood and the nervous tissue. Our aim was to develop and characterize a highly reproducible rat syngeneic in vitro model of the BBB using co-cultures of primary rat brain endothelial cells (RBEC) and astrocytes to study receptors involved in transcytosis across the endothelial cell monolayer. Astrocytes were isolated by mechanical dissection following trypsin digestion and were frozen for later co-culture. RBEC were isolated from 5-week-old rat cortices. The brains were cleaned of meninges and white matter, and mechanically dissociated following enzymatic digestion. Thereafter, the tissue homogenate was centrifuged in bovine serum albumin to separate vessel fragments from nervous tissue. The vessel fragments underwent a second enzymatic digestion to free endothelial cells from their extracellular matrix. The remaining contaminating cells such as pericytes were further eliminated by plating the microvessel fragments in puromycin-containing medium. They were then passaged onto filters for co-culture with astrocytes grown on the bottom of the wells. RBEC expressed high levels of tight junction (TJ) proteins such as occludin, claudin-5 and ZO-1 with a typical localization at the cell borders. The transendothelial electrical resistance (TEER) of brain endothelial monolayers, indicating the tightness of TJs reached 300 ohm x cm(2) on average. The endothelial permeability coefficients (Pe) for lucifer yellow (LY) was highly reproducible with an average of 0.26 ± 0.11 x 10(-3) cm/min. Brain endothelial cells organized in monolayers expressed the efflux transporter P-glycoprotein (P-gp), showed a polarized transport of rhodamine 123, a ligand for P-gp, and showed specific transport of transferrin-Cy3 and DiILDL across the endothelial cell monolayer. In conclusion, we provide a protocol for setting up an in vitro BBB model that is highly reproducible due to the quality assurance methods, and that is suitable for research on BBB transporters and receptors.


A combination of metformin and galantamine exhibits synergistic benefits in the treatment of sarcopenia.
Age-associated sarcopenia, characterized by a progressive loss in muscle mass and strength, is the largest cause of frailty and disability in the elderly worldwide. Current treatments involve nonpharmacological guidelines that few subjects can abide by, highlighting the need for effective drugs. Preclinical models were employed to test the benefits of RJx-01, a combination drug composed of metformin and galantamine, on sarcopenia. In worms, RJx-01 treatment improved lifespan, locomotion, pharyngeal pumping, and muscle fiber organization. The synergistic effects of RJx-01 were recapitulated in a transgenic mouse model that displays an exacerbated aging phenotype (Opa1-/-). In these mice, RJx-01 ameliorated physical performance, muscle mass and force, neuromuscular junction stability, and systemic inflammation. RJx-01 also improved physical performance and muscle strength in 22-month-old WT mice and also improved skeletal muscle ultrastructure, mitochondrial morphology, autophagy, lysosomal function, and satellite cell content. Denervation and myofiber damage were decreased in RJx-01-treated animals compared with controls. RJx-01 improved muscle quality rather than quantity, indicating that the improvement in quality underlies the beneficial effects of the combination drug. The studies herein indicate synergistic beneficial effects of RJx-01 in the treatment of sarcopenia and support the pursuit of RJx-01 in a human clinical trial as a therapeutic intervention for sarcopenia.


Kardiovize Brno 2030, a prospective cardiovascular health study in Central Europe: Methods, baseline findings and future directions.
Background Atherosclerotic cardiovascular disease is highly prevalent in Eastern and Central Europe, where the incidence is the highest in the world. The Kardiovize Brno 2030 study was designed as a prospective cohort study to investigate the complex relationships of cardiovascular disease and outcomes with a range of biological, psychosocial, environmental, behavioral, and economic factors in an urban population of the Czech Republic. Methods We randomly selected a 1% sample of the city of Brno residents aged 25-64 years stratified by sex and age. The study assessed traditional and novel cardiovascular disease risk factors, including sociodemographic and smoking status, physical activity, diet, depression, stress, body fat, cardio-ankle vascular index, and intima media thickness, complemented by blood tests; biological samples were stored for future analyses. Results The study enrolled 2160 participants (54.8% women), with a mean age of 47 ± 11.3 years. They were mostly full-time employed (75.6%) and married (62.1%). Hyperlipidemia was highly prevalent (70.7% in men, and 67.1% in women, NS). Hypertension and diabetes mellitus were more prevalent in men than in women (54.3% vs. 38.7% and 7.1% vs. 3.5%, respectively, P < 0.001 for both). A total of 25.3% of men and 21.9% of women smoked, whereas 20.0% and 43.0% of men and 18.1% and 26.6% of women were obese and overweight, respectively. Conclusions Cardiovascular risk factors are highly prevalent in the city of Brno, an urban population from Central Europe. The Kardiovize Brno 2030 study will provide unique multidimensional and longitudinal cardiovascular health data from a region where epidemiological studies are scarce.


Measuring preschool learning engagement in the laboratory.
Learning engagement is a critical factor for academic achievement and successful school transitioning. However, current methods of assessing learning engagement in young children are limited to teacher report or classroom observation, which may limit the types of research questions one could assess about this construct. The current study investigated the validity of a novel assessment designed to measure behavioral learning engagement among young children in a standardized laboratory setting and examined how learning engagement in the laboratory relates to future classroom adjustment. Preschool-aged children (N = 278) participated in a learning-based Tangrams task and Story sequencing task and were observed based on seven behavioral indicators of engagement. Confirmatory factor analysis supported the construct validity for a behavioral engagement factor composed of six of the original behavioral indicators: attention to instructions, on-task behavior, enthusiasm/energy, persistence, monitoring progress/strategy use, and negative affect. Concurrent validity for this behavioral engagement factor was established through its associations with parent-reported mastery motivation and pre-academic skills in math and literacy measured in the laboratory, and predictive validity was demonstrated through its associations with teacher-reported classroom learning behaviors and performance in math and reading in kindergarten. These associations were found when behavioral engagement was observed during both the nonverbal task and the verbal story sequencing tasks and persisted even after controlling for child minority status, gender, and maternal education. Learning engagement in preschool appears to be successfully measurable in a laboratory setting. This finding has implications for future research on the mechanisms that support successful academic development.


Isolation of a small molecule inhibitor of DNA base excision repair.
The base excision repair (BER) pathway is essential for the removal of DNA bases damaged by alkylation or oxidation. A key step in BER is the processing of an apurinic/apyrimidinic (AP) site intermediate by an AP endonuclease. The major AP endonuclease in human cells (APE1, also termed HAP1 and Ref-1) accounts for >95% of the total AP endonuclease activity, and is essential for the protection of cells against the toxic effects of several classes of DNA damaging agents. Moreover, APE1 overexpression has been linked to radio- and chemo-resistance in human tumors. Using a newly developed high-throughput screen, several chemical inhibitors of APE1 have been isolated. Amongst these, CRT0044876 was identified as a potent and selective APE1 inhibitor. CRT0044876 inhibits the AP endonuclease, 3'-phosphodiesterase and 3'-phosphatase activities of APE1 at low micromolar concentrations, and is a specific inhibitor of the exonuclease III family of enzymes to which APE1 belongs. At non-cytotoxic concentrations, CRT0044876 potentiates the cytotoxicity of several DNA base-targeting compounds. This enhancement of cytotoxicity is associated with an accumulation of unrepaired AP sites. In silico modeling studies suggest that CRT0044876 binds to the active site of APE1. These studies provide both a novel reagent for probing APE1 function in human cells, and a rational basis for the development of APE1-targeting drugs for antitumor therapy.