Sequence-specific recognition of a PxLPxI/L motif by an ankyrin repeat tumbler lock.
Ankyrin repeat family A protein 2 (ANKRA2) interacts with the plasma membrane receptor megalin and the class IIa histone deacetylases HDAC4 and HDAC5. We report that the ankyrin repeat domains of ANKRA2 and its close paralog regulatory factor X-associated ankyrin-containing protein (RFXANK) recognize a PxLPxI/L motif found in diverse binding proteins, including HDAC4, HDAC5, HDAC9, megalin, and regulatory factor X, 5 (RFX5). Crystal structures of the ankyrin repeat domain of ANKRA2 in complex with its binding peptides revealed that each of the middle three ankyrin repeats of ANKRA2 recognizes a residue from the PxLPxI/L motif in a tumbler-lock binding mode, with ANKRA2 acting as the lock and the linear binding motif serving as the key. Structural analysis showed that three disease-causing mutations in RFXANK affect residues that are critical for binding to RFX5. These results suggest a fundamental principle of longitudinal recognition of linear sequences by a repeat-type domain. In addition, phosphorylation of serine 350, a residue embedded within the PxLPxI/L motif of HDAC4, impaired the binding of ANKRA2 but generated a high-affinity docking site for 14-3-3 proteins, which may help sequester this HDAC in the cytoplasm. Thus, the binding preference of the PxLPxI/L motif is signal-dependent. Furthermore, proteome-wide screening suggested that a similar phosphorylation-dependent switch may operate in other pathways. Together, our findings uncover a previously uncharacterized sequence- and signal-dependent peptide recognition mode for a repeat-type protein domain.


Effects of lamb sucking on the bacterial flora of teat duct and mammary gland of ewes.
We aimed (i) to determine differences in bacterial flora of teat duct and mammary gland of ewes before and after suckling, (ii) to evaluate factors potentially affecting those. We collected samples of teat duct material and mammary secretion from 11 ewes immediately before and after sucking by lambs, as well as 120 min later. We processed samples bacteriologically and compared changes in infection by the Sign Test. We isolated bacteria from 3.5% duct and 1.5% secretion samples before suckling. Respective figures post-suckling were 10.6% and 2.0%, and 120 min later 6.8% and 1.5%. We recorded differences in infection of duct samples before and after suckling in 40 cases; bacteria were isolated before suckling from six samples, whereas after it from 34 (p < 0.001). Also, we recorded differences in samples collected after suckling and 120 min later in 12 cases; bacteria were isolated immediately post-suckling from eight samples, whereas 120 min later from four (p = 0.375). No significant changes were seen for secretion. We found neither difference between ewes with single or twin lambs, nor among stages of lactation. Mostly, we isolated staphylococci: 70% of isolates before suckling, 80% of isolates after it, 91% of isolates 120 min later. After suckling we also isolated two Mannheimia haemolytica strains. Suckling predisposes to entrance of bacteria into the teat; however, increased teat infections did not result in mammary infections. Isolation of M. haemolytica post-suckling indicates that lambs act as source of infection for this pathogen.


Molecular iron(III) phosphonates: synthesis, structure, magnetism, and Mössbauer studies.
The reaction of Fe(ClO4)2·6H2O with t-BuPO3H2 or Cl3CPO3H2 in the presence of an ancillary pyrazole phenolate as a coligand, H2phpzH [H2phpzH = 3(5)-(2-hydroxyphenyl)pyrazole], afforded tetra- and pentanuclear Fe(III) phosphonate complexes [Fe4(t-BuPO3)4(HphpzH)4]·5CH3CN·5CH2Cl2 (1) and [HNEt3]2[Fe5(μ3-O)(μ-OH)2 (Cl3CPO3)3(HphpzH)5(μ-phpzH]·3CH3CN·2H2O (2). Single-crystal X-ray structural analysis reveals that 1 possesses a cubic double-4-ring (D4R) core similar to what is found in zeolites. The molecular structure of 2 reveals it to be pentanuclear. It crystallizes in the chiral P1 space group. Magnetic studies on 1 and 2 have also been carried out, which reveal that the bridging phosphonate ligands mediate weak antiferromagnetic interactions between the Fe(III) ions. Magnetization dynamics of 1 and 2 have been corroborated by a Mössbauer spectroscopy analysis.


Problem-based medical education: development of a theoretical foundation and a science-based professional attitude.
Problem-based learning, combined with early patient contact, integration between different subject areas, elements of multiprofessional education, and special emphasis on the development of communications skills has become the basis for the medical curriculum at the Faculty of Health Sciences in Linköping. Critics have questioned the depth of the scientific and theoretical aspects of the curriculum. Through a series of specific measures in the organization of the curriculum and examinations, and due to the pedagogical principles involved per se, our claim is that students graduating at Linköping do possess the required theoretical knowledge and a scientific attitude to the practice of medicine, at least equivalent to that obtained in a more conventional medical curriculum. One such specific measure is that all students perform one field study and two scientific studies during the course of the curriculum. An investigation of student opinions regarding the value of performing scientific projects of their own have shown that these projects have had a positive impact on the students' general scientific attitude and their willingness to engage in future scientific work. The specific skills acquired, as confirmed by oral examinations, were largely determined by the scientific nature of the chosen field of study. Our graduates have not yet progressed far enough in their careers for comparisons to be made on the basis of the Swedish Licensing Board Internship Examinations, but continuing evaluations of students, graduates and licensed doctors emerging from the curriculum will provide future evidence as to whether our present evaluation is correct.


Gene Therapy for Pediatric Cancer: State of the Art and Future Perspectives.
While modern treatments have led to a dramatic improvement in survival for pediatric malignancy, toxicities are high and a significant proportion of patients remain resistant. Gene transfer offers the prospect of highly specific therapies for childhood cancer. "Corrective" genes may be transferred to overcome the genetic abnormalities present in the precancerous cell. Alternatively, genes can be introduced to render the malignant cell sensitive to therapeutic drugs. The tumor can also be attacked by decreasing its blood supply with genes that inhibit vascular growth. Another possible approach is to modify normal tissues with genes that make them more resistant to conventional drugs and/or radiation, thereby increasing the therapeutic index. Finally, it may be possible to attack the tumor indirectly by using genes that modify the behavior of the immune system, either by making the tumor more immunogenic, or by rendering host effector cells more efficient. Several gene therapy applications have already been reported for pediatric cancer patients in preliminary Phase 1 studies. Although no major clinical success has yet been achieved, improvements in gene delivery technologies and a better understanding of mechanisms of tumor progression and immune escape have opened new perspectives for the cure of pediatric cancer by combining gene therapy with standard therapeutic available treatments.


Visible light-induced insulin aggregation on surfaces via photoexcitation of bound thioflavin T.
Insulin is known to form amyloid aggregates when agitated in a hydrophobic container. Amyloid aggregation is routinely measured by the fluorescence of the conformational dye thioflavin T, which, when incorporated into amyloid fibers, fluoresces at 480 nm. The kinetics of amyloid aggregation in general is characterized by an initial lag-phase, during which aggregative nuclei form on the hydrophobic surface. These nuclei then lead to the formation of fibrils presenting a rapid growth during the elongation phase. Here we describe a novel mechanism of insulin amyloid aggregation which is surprisingly devoid of a lag-time for nucleation. The excitation of thioflavin T by visible light at 440 nm induces the aggregation of thioflavin T-positive insulin fibrils on hydrophobic surfaces in the presence of strong agitation and at physiological pH. This process is material surface-induced and depends on the fact that surface-adsorbed insulin can bind thioflavin T. Light-induced insulin aggregation kinetics is thioflavin T-mediated and is based on an energy transfer from visible light to the protein via thioflavin T. It relies on a constant supply of thioflavin T and insulin from the solution to the aggregate. The growth rate increases with the irradiance and with the concentration of thioflavin T. The supply of insulin seems to be the limiting factor of aggregate growth. This light-induced aggregation process allows the formation of local surface-bound aggregation patterns.


Imaging of the vulnerable carotid plaque: biological targeting of inflammation in atherosclerosis using iron oxide particles and MRI.
Identification of those patients with high-risk asymptomatic carotid plaques remains an elusive but essential step in stroke prevention. Inflammation is a key process in plaque destabilization and the propensity of atherosclerotic lesions to cause clinical sequelae. There is currently no clinical imaging technique available to assess the degree of inflammation associated with plaques. This study aims at visualizing and characterizing atherosclerosis using antibody-conjugated superparamagnetic iron oxide (SPIO) particles as an MRI probe to assess inflammation in human atherosclerotic plaques. Atherosclerotic plaques were collected from 20 consecutive patients (n=10 from symptomatic patients, n=10 from asymptomatic patients) undergoing carotid endarterectomy (CEA) for extracranial high-grade internal carotid artery (ICA) stenosis (>70% luminal narrowing). Inflammatory markers on human atherosclerotic plaques were detected and characterized by ex vivo magnetic resonance imaging (MRI) using anti-VCAM-1 antibody and anti-E-selectin antibody-conjugated SPIO with confirmatory immunohistochemistry. Inflammation associated with human ex vivo atherosclerotic plaques could be imaged using dual antibody-conjugated SPIO by MRI. Symptomatic plaques could be distinguished from asymptomatic ones by the degree of inflammation, and the MR contrast effect was significantly correlated with the degree of plaque inflammation (r=.64, p<.001). The asymptomatic plaque population exhibited heterogeneity in terms of inflammation. The dual-targeted SPIO-induced MR signal not only tracked closely with endothelial activation (i.e. endothelial expression of VCAM-1 and E-selectin), but also reflected the macrophage burden within plaque lesions, offering a potential imaging tool for quantitative MRI of inflammatory activity in atherosclerosis. These functional molecular MRI probes constitute a novel imaging tool for ex vivo characterization of atherosclerosis at a molecular level. Further development and translation into the clinical arena will facilitate more accurate risk stratification in carotid artery disease in the future.


[Alcohol consumption in pregnancy: performance of the Brazilian version of the questionnaire T-ACE].
To assess the performance characteristics of the Brazilian version of the Tolerance, Annoyed, Cut down and Eye-opener (T-ACE) questionnaire to screen alcohol consumption during pregnancy. Observational, cross-sectional study in a sequential sample of 450 women in the third trimester of pregnancy, attended in a maternity ward in a city of Southeastern Brazil, in 2001. The following instruments were used: a questionnaire to gather sociodemographic data, the T-ACE, a questionnaire to verify history of alcohol consumption throughout gestation, and a clinical interview to identify the harmful use of and dependence on alcohol, according to ICD-10 diagnostic criteria. Concordance tests among different interviewers as well as test-/re-test reliability tests were performed. A total of 100 women (22.1%) were identified as positive by the T-ACE. The kappa indexes for concordance and reliability were 0.95, with 97% of concordant responses. When compared to the ICD-10 criteria and to the pattern of consumption, the T-ACE, with a cut-off point of two or higher, presented sensitivity and specificity coefficients of 100% and 85%, and of 97.9% and 86.6%, respectively. The Brazilian version of the T-ACE seemed to appropriately meet the performance criteria that qualify it as a basic instrument for the screening of alcohol consumption during pregnancy. Its use in the routine and practice of obstetric services is recommended in view of the tendency for increased alcohol consumption among women, the difficulties to identify alcohol abuse by pregnant women, and the risk of developmental problems in children.


When trends intersect: The challenge of protecting freshwater ecosystems under multiple land use and hydrological intensification scenarios.
Intensification of the use of natural resources is a world-wide trend driven by the increasing demand for water, food, fibre, minerals and energy. These demands are the result of a rising world population, increasing wealth and greater global focus on economic growth. Land use intensification, together with climate change, is also driving intensification of the global hydrological cycle. Both processes will have major socio-economic and ecological implications for global water availability. In this paper we focus on the implications of land use intensification for the conservation and management of freshwater ecosystems using Australia as an example. We consider this in the light of intensification of the hydrologic cycle due to climate change, and associated hydrological scenarios that include the occurrence of more intense hydrological events (extreme storms, larger floods and longer droughts). We highlight the importance of managing water quality, the value of providing environmental flows within a watershed framework and the critical role that innovative science and adaptive management must play in developing proactive and robust responses to intensification. We also suggest research priorities to support improved systemic governance, including adaptation planning and management to maximise freshwater biodiversity outcomes while supporting the socio-economic objectives driving land use intensification. Further research priorities include: i) determining the relative contributions of surface water and groundwater in supporting freshwater ecosystems; ii) identifying and protecting freshwater biodiversity hotspots and refugia; iii) improving our capacity to model hydro-ecological relationships and predict ecological outcomes from land use intensification and climate change; iv) developing an understanding of long term ecosystem behaviour; and v) exploring systemic approaches to enhancing governance systems, including planning and management systems affecting freshwater outcomes. A major policy challenge will be the integration of land and water management, which increasingly are being considered within different policy frameworks.


A Kenyan perspective on the use of animals in science education and scientific research in Africa and prospects for improvement.
Animal experimentation is common in Africa, a region that accords little priority on animal protection in comparison to economic and social development. The current study aimed at investigating the prevalence of animal experimentation in Kenya, and to review shortfalls in policy, legislation, implementation and enforcement that result in inadequate animal care in Kenya and other African nations. Data was collected using questionnaires, administered at 39 highly ranked academic and research institutions aiming to identify those that used animals, their sources of animals, and application of the three Rs. Perceived challenges to the use of non-animal alternatives and common methods of euthanasia were also queried. Data was analyzed using Epidata, SPSS 16.0 and Microsoft Excel. Thirty-eight (97.4%) of thirty-nine institutions reported using animals for education and/or research. Thirty (76.9%) institutions reported using analgesics or anesthetics on a regular basis. Thirteen (33.3%) institutions regularly used statistical methods to minimize the use of animals. Overall, sixteen (41.0%) institutions explored the use of alternatives to animals such as cell cultures and computer simulation techniques, with one (2.6%) academic institution having completely replaced animals with computer modeling, manikins and visual illustrations. The commonest form of euthanasia employed was chloroform administration, reportedly in fourteen (29.8%) of 47 total methods (some institutions used more than one method). Twenty-eight (71.8%) institutions had no designated ethics committee to review or monitor protocols using animals. Animals are commonly used in academic and research institutions in Kenya. The relative lack of ethical guidance and oversight regarding the use of animals in research and education presents significant concerns.


Merging and characterising phenotypic data on conventional and rare traits from dairy cattle experimental resources in three countries.
This study set out to demonstrate the feasibility of merging data from different experimental resource dairy populations for joint genetic analyses. Data from four experimental herds located in three different countries (Scotland, Ireland and the Netherlands) were used for this purpose. Animals were first lactation Holstein cows that participated in ongoing or previously completed selection and feeding experiments. Data included a total of 60 058 weekly records from 1630 cows across the four herds; number of cows per herd ranged from 90 to 563. Weekly records were extracted from the individual herd databases and included seven traits: milk, fat and protein yield, milk somatic cell count, liveweight, dry matter intake and energy intake. Missing records were predicted with the use of random regression models, so that at the end there were 44 weekly records, corresponding to the typical 305-day lactation, for each cow. A total of 23 different lactation traits were derived from these records: total milk, fat and protein yield, average fat and protein percentage, average fat-to-protein ratio, total dry matter and energy intake and average dry matter intake-to-milk yield ratio in lactation weeks 1 to 44 and 1 to 15; average milk somatic cell count in lactation weeks 1 to 15 and 16 to 44; average liveweight in lactation weeks 1 to 44; and average energy balance in lactation weeks 1 to 44 and 1 to 15. Data were subsequently merged across the four herds into a single dataset, which was analysed with mixed linear models. Genetic variance and heritability estimates were greater (P < 0.05) than zero for all traits except for average milk somatic cell count in weeks 16 to 44. Proportion of total phenotypic variance due to genotype-by-environment (sire-by-herd) interaction was not different (P > 0.05) from zero. When estimable, the genetic correlation between herds ranged from 0.85 to 0.99. Results suggested that merging experimental herd data into a single dataset is both feasible and sensible, despite potential differences in management and recording of the animals in the four herds. Merging experimental data will increase power of detection in a genetic analysis and augment the potential reference population in genome-wide association studies, especially of difficult-to-record traits.


Provoked flux motion of cochlear blood flow measured with laser Doppler flowmetry in guinea pig.
Although progress has been made in the study of cochlear blood flow (CBF) regulation since laser Doppler flowmetry (LDF) was introduced, cochlear vasomotion has not been investigated. Therefore, the primary objective of this study was to determine if oscillatory fluctuations of CBF could be provoked. Guinea pigs were anesthetized with diazepam (5 mg/kg) and fentanyl (0.32 mg/kg). Blood pressure (BP) was recorded from a carotid artery cannula. The cochlea and pons cerebellum were ventrally exposed; the bilateral CBF and brain blood flow (BBF) or skin blood flow (SBF) were monitored by LDF. After administration of phentolamine (0.25-0.75 mg/kg, i.v.), ipsilateral CBF in 7 of 16 animals showed a 2-5 min episode of oscillation. During artificial hyperventilation, continuous oscillation of CBF was recorded (the flux motion frequency was 3.5 +/- 0.5 cycles per min and its amplitude 25.8 +/- 5.6% from baseline). The time-dependent flux change (the waveform) was the same throughout a single cochlea but different between cochleae of the same animal. Compared to BBF, CBF vasomotion frequency was lower, and amplitude larger. SBF exhibited no such motion. Flux motion could be eliminated by inhalation of pure oxygen or 5% CO2 in oxygen or by the smooth muscle relaxants, papaverine and hydralazine. Phentolamine-induced vasomotion may be due to a hypotensive perfusion pressure, and hyperventilation-enhanced vasomotion may be caused by changing blood gas concentrations and by hormonal or neuronal activity. Oxygen and CO2 inhalation slightly increased BP and this change in perfusion pressure was probably associated with weakened vasomotion.(ABSTRACT TRUNCATED AT 250 WORDS)


anti-TNF agents as therapeutic choice in immune-mediated inflammatory diseases: focus on adalimumab.
The complex pathogenesis of immune-mediated inflammatory diseases (IMIDs) has been extensively investigated and dysregulation of cytokines, such as tumour necrosis factor (TNF) has been shown to play a dominant role in the pathogenesis of various IMIDs, such as rheumatoid arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, psoriasis and psoriatic arthritis. The subsequent development of biological agents capable of blocking TNF has led to important advances in the pharmacotherapy of such diseases and confirmed the concept of a common pathophysiology among IMIDs with TNF having a predominant role. Five TNF inhibitors have currently been approved for treatment of one or more IMIDs; these include infliximab, etanercept, adalimumab, golimumab and certolizumab pegol. Given the similarities in the pathogenic background of IMIDs, one could expect that anti-TNF agents be similarly effective and with comparable tolerability profiles; however, this may not be the case. Structural and pharmacological differences among the anti-TNF drugs are likely to result in differences in efficacy and tolerability among the agents in the different IMIDs, together with differences in potency, therapeutic dose ranges, dosing regimens, administration routes, and propensity for immunogenicity. Among the five TNF inhibitors approved for treatment of IMIDs, adalimumab has the widest range of indications. Data from controlled clinical trials of adalimumab, showing its excellent efficacy and tolerability in a wide range of indications, are supported by real-world long-term data from observational studies, which confirm the value of adalimumab as a suitable choice in the management of IMIDs.


What influences the awareness of physician quality information? Implications for Medicare.
Examine the factors that are associated with awareness of physician quality information (PQI) among older people with one or more chronic illnesses and the implications for Medicare. Random digit-dial survey of adults with one or more chronic illnesses. Structural equation modeling to examine factors related to awareness of PQI. Awareness of PQI is low (13 percent), but comparable to findings in general population surveys. Age, race, education, and self-reported health status are associated with PQI awareness. Trust in the Internet as a source of health care information and not trusting one's physician as a source of information both are associated with a greater likelihood of being aware of PQI. Patients with high levels of activation have greater trust in physicians as information sources, but this is not associated with awareness, nor is degree of satisfaction with their care experience. Awareness of PQI among older persons with chronic illnesses is relatively low across all socio-economic and demographic subgroups. Changes in population characteristics over time are unlikely to improve awareness in this population, nor are changes in patient activation or satisfaction with care. Medicare would need a broad-based effort if it wishes to raise PQI awareness among Medicare beneficiaries in the near term. Before undertaking resource-intensive efforts to increase awareness, Medicare may want to consider what level of awareness actually is needed to accomplish the overall objective for PQI transparency, which is raising the quality of care received by beneficiaries. It may be that relatively low levels of awareness are sufficient.


Stimulation of RNA polymerase II elongation by hepatitis delta antigen.
Transcription elongation by RNA polymerase II (RNAPII) is negatively regulated by the human factors DRB-sensitivity inducing factor (DSIF) and negative elongation factor (NELF). A 66-kilodalton subunit of NELF (NELF-A) shows limited sequence similarity to hepatitis delta antigen (HDAg), the viral protein required for replication of hepatitis delta virus (HDV). The host RNAPII has been implicated in HDV replication, but the detailed mechanism and the role of HDAg in this process are not understood. We show that HDAg binds RNAPII directly and stimulates transcription by displacing NELF and promoting RNAPII elongation. These results suggest that HDAg may regulate RNAPII elongation during both cellular messenger RNA synthesis and HDV RNA replication.


Role of CCR5 in infection of primary macrophages and lymphocytes by macrophage-tropic strains of human immunodeficiency virus: resistance to patient-derived and prototype isolates resulting from the delta ccr5 mutation.
The alpha-chemokine receptor fusin (CXCR-4) and beta-chemokine receptor CCR5 serve as entry cofactors for T-cell (T)-tropic and macrophage (M)-tropic human immunodeficiency virus type 1 (HIV-1) strains, respectively, when expressed with CD4 in otherwise nonpermissive cells. Some M-tropic and dual-tropic strains can also utilize other beta-chemokine receptors, such as CCR2b and CCR3. A mutation of CCR5 (delta ccr5) was recently found to be common in certain populations and appears to confer protection against HIV-1 in vivo. Here, we show that this mutation results in a protein that is expressed intracellularly but not on the cell surface. Primary CD4 T cells from delta ccr5 homozygous individuals were highly resistant to infection with prototype M-tropic HIV-1 strains, including an isolate (YU-2) that uses CCR5 and CCR3, but were permissive for both a T-tropic strain (3B) and a dual-tropic variant (89.6) that uses CXCR-4, CCR5, CCR3, or CCR2b. These cells were also resistant to M-tropic patient isolates but were readily infected by T-tropic patient isolates. Primary macrophages from delta ccr5 homozygous individuals were also resistant to infection with M-tropic strains, including YU-2, but the dual-tropic strain 89.6 was able to replicate in them even though macrophages are highly resistant to CXCR-4-dependent T-tropic isolates. These data show that CCR5 is the essential cofactor for infection of both primary macrophages and T lymphocytes by most M-tropic strains of HIV-1. They also suggest that CCR3 does not function for HIV-1 entry in primary lymphocytes or macrophages, but that a molecule(s) other than CCR5 can support entry into macrophages by certain virus isolates. These studies further define the cellular basis for the resistance to HIV-1 infection of individuals lacking functional CCR5.


The accuracy of fine-needle aspiration cytology for diagnosis of parotid gland masses: a clinicopathological study of 114 patients.
Fine-needle aspiration cytology is a valuable method for preoperative assessment of head and neck tumors. However, its accuracy in detection of salivary gland masses is controversial compared with other methods. The aim of this work was to evaluate the effectiveness and accuracy of fine-needle aspiration cytology (FNAC) in the diagnosis of parotid gland masses. Over a 10-year period, 126 parotid gland masses were resected. Retrospective chart reviews of 114 patients were performed. The results of FNAC and final histological diagnosis were compared and the accuracy of FNAC was determined. Final histological evaluation revealed 11 malignant tumors and 103 benign lesions. Pleomorphic adenoma was the most common neoplasm (63%), followed by Warthin's tumor (17.5%). The sensitivity of FNAC in detecting malignant tumors was 73% and the specificity was 97%. Positive predictive value (PPV) was 73% and negative predictive value (NPV) was 97%. The overall accuracy of FNAC in detecting parotid masses was 95%. False-negative diagnosis was found in mucoepidermoid carcinoma, acinic cell carcinoma, and epithelial-myoepithelial carcinoma whereas there was false-positive diagnosis in cases of pleomorphic adenoma and normal parotid gland tissue. FNAC is a reliable minimally invasive diagnostic method with a high sensitivity in diagnosis of lesions in parotid glands. The sensitivity of detection of malignant tumors in parotid glands was low due to the biopsy technique used, and depended on tumor location. Postoperative complications decreased after superficial parotidectomy.


Wolbachia association with the tsetse fly, Glossina fuscipes fuscipes, reveals high levels of genetic diversity and complex evolutionary dynamics.
Wolbachia pipientis, a diverse group of α-proteobacteria, can alter arthropod host reproduction and confer a reproductive advantage to Wolbachia-infected females (cytoplasmic incompatibility (CI)). This advantage can alter host population genetics because Wolbachia-infected females produce more offspring with their own mitochondrial DNA (mtDNA) haplotypes than uninfected females. Thus, these host haplotypes become common or fixed (selective sweep). Although simulations suggest that for a CI-mediated sweep to occur, there must be a transient phase with repeated initial infections of multiple individual hosts by different Wolbachia strains, this has not been observed empirically. Wolbachia has been found in the tsetse fly, Glossina fuscipes fuscipes, but it is not limited to a single host haplotype, suggesting that CI did not impact its population structure. However, host population genetic differentiation could have been generated if multiple Wolbachia strains interacted in some populations. Here, we investigated Wolbachia genetic variation in G. f. fuscipes populations of known host genetic composition in Uganda. We tested for the presence of multiple Wolbachia strains using Multi-Locus Sequence Typing (MLST) and for an association between geographic region and host mtDNA haplotype using Wolbachia DNA sequence from a variable locus, groEL (heat shock protein 60). MLST demonstrated that some G. f. fuscipes carry Wolbachia strains from two lineages. GroEL revealed high levels of sequence diversity within and between individuals (Haplotype diversity = 0.945). We found Wolbachia associated with 26 host mtDNA haplotypes, an unprecedented result. We observed a geographical association of one Wolbachia lineage with southern host mtDNA haplotypes, but it was non-significant (p = 0.16). Though most Wolbachia-infected host haplotypes were those found in the contact region between host mtDNA groups, this association was non-significant (p = 0.17). High Wolbachia sequence diversity and the association of Wolbachia with multiple host haplotypes suggest that different Wolbachia strains infected G. f. fuscipes multiple times independently. We suggest that these observations reflect a transient phase in Wolbachia evolution that is influenced by the long gestation and low reproductive output of tsetse. Although G. f. fuscipes is superinfected with Wolbachia, our data does not support that bidirectional CI has influenced host genetic diversity in Uganda.


Lack of interaction between devazepide and 8-OH-DPAT-induced hyperphagia in the rat.
Recently, a number of studies have provided evidence suggesting that CCK and 5-HT interact in the control of food intake. However, the majority of these studies have relied on the administration of exogenous CCK to investigate potential interactions. The aim of the present study was to focus on the potential role of endogenous CCK in 5-HT-CCK interactions. Our prediction was that the CCKA antagonist, devazepide, alone would potentiate the hyperphagic effect of the 5-HT1A agonist, 8-OH-DPAT, in free-feeding rats. The results showed that devazepide, at a dose that had no intrinsic effect (1.0 mg/kg), did not enhance the hyperphagic effect of 8-OH-DPAT (100 and 300 micrograms/kg). This suggests that when serotonergic inhibitory activity is reduced by 5-HT1A-receptor stimulation, there is no compensatory increase of endogenous CCK activity to excite 5-HT neurons and thereby inhibit food intake.


Fingerstick Helicobacter pylori antibody test: better than laboratory serological testing?
Antibody testing is the recommended method to screen for Helicobacter pylori (H. pylori) infection. Whole-blood fingerstick antibody tests are simple, in-office tests providing rapid results, but the accuracy of first-generation tests was lower than other diagnostic tests. We assessed a new whole-blood antibody test, using endoscopic biopsy tests as a "gold standard," and compared it with a laboratory quantitative serological test. Two hundred-one patients not previously treated for H. pylori who were undergoing endoscopy had gastric biopsies for rapid urease test and histological examination; whole-blood antibody tests and quantitative serological tests were also performed. Two separate gold standards for H. pylori infection were employed: either rapid urease test or histological exam positive; and both rapid urease test and histological exam positive. Sensitivities for whole-blood test versus quantitative serology with gold standard 1 (either biopsy test positive) were 86% versus 92% (95% confidence interval [CI] of difference, -2-14%; p = 0.19) and specificities were 88% versus 77% (95% CI of difference, 0.4-22%; p = 0.052). Sensitivities with gold standard 2 (both biopsy tests positive) were 90% versus 94% (95% CI of difference, -4-12%; p = 0.41) and specificities were 79% versus 67% (95% CI of difference, 1-24%; p = 0.048). New generation in-office, whole-blood antibody tests that can achieve a sensitivity and specificity similar to or better than those of widely used quantitative laboratory serological tests may be used as the initial screening tests of choice for H. pylori.


A functional study on CysLT(1) receptors in human eosinophils.
The cysteinyl leukotrienes (CysLTs) mediate their biological actions through two receptors: CysLT(1) receptor and CysLT(2) receptor. This study was undertaken to examine the direct effects of CysLTs on eosinophils, such as chemotaxis and degranulation, focusing on CysLT(1). Eosinophils were isolated from venous blood from normal volunteers who had no history of allergy (purity >99%). They were subjected to reverse transcription-PCR analysis and flow-cytometric analysis for CysLT(1). Binding assays were performed with [(3)H]LTD(4). Purified eosinophils loaded with Fura-2 acetoxymethyl ester were stimulated with CysLTs, and Ca(2+) influx was measured. Eosinophil migration in response to CysLTs was measured using a 96-well multiwell Boyden chamber. Eosinophils were treated with LTD(4) at 10(-6) M for 60 min followed by incubation for 4 h at 37 degrees C in the presence or absence of IL-5 and eosinophil-derived neurotoxin (EDN) release was evaluated. The expression of the mRNA and protein of CysLT(1) on eosinophils and [(3)H]LTD(4)-specific binding to eosinophils were observed. Neither Th1 cytokine (IFN-gamma) nor Th2 cytokines (IL-4 or IL-5) affected CysLT(1) expression in eosinophils. CysLTs induced an increase in intracellular free Ca(2+) in eosinophils via CysLT(1), as suggested by the efficient inhibition by a CysLT(1) antagonist, pranlukast, in addition to the rank order of potency being LTD(4), LTC(4) and LTE(4). LTD(4) stimulated eosinophils to migrate at 10(-6) M via CysLT(1). LTE(4) also induced significant eosinophil migration at 10(-6) M. LTD(4) enhanced EDN release induced by IL-5 via CysLT(1). CysLTs induce migration and enhance degranulation in eosinophils via CysLT(1). Accordingly, interaction of CysLTs and CysLT(1) on eosinophils has the potential to play a prominent role in the pathophysiology of asthma.


Antileishmanial efficacy of Boerhaavia diffusa L. and Ocimum sanctum L. against experimental visceral leishmaniasis.
The chemotherapy of visceral leishmaniasis (VL) has several limitations including resistance and toxicity of the existing drugs. Down regulation of immune system further aggravates the problems. To combat this situation we evaluated the leishmanicidal efficacy of Boerhaavia diffusa and Ocimum sanctum through oral route in L. donovani infection in BALB/c mice. Results have demonstrated maximum clearance of the parasites from infected animals treated with combination of B. diffusa and O. sanctum (@ 100 and 400 mg/kg body wt., respectively 5 days) as depicted through Leishman Donovan Units in liver. Up-regulation of cell-mediated immunity was also observed in animals of this group as heightened delayed type hypersensitivity responses and increased IgG2a levels were observed. Moreover, increased levels of SGOT, SGPT, serum urea, blood urea nitrogen and serum creatinine were brought down to normal levels. Since VL is associated immunosuppression, the above treatment is a good option as it helps in the up-regulation of Th1 responses and reduction in parasite load in L. donovani infected mice. These findings suggest a new option for antileishmanial chemotherapy at lower cost and nil toxicity.


Gastrointestinal neuroendocrine tumors.
Gastrointestinal neuroendocrine tumors (GI-NETs) are a genetically diverse group of malignancies that sometimes produce peptides causing characteristic hormonal syndromes. NETs can be clinically symptomatic (functioning) or silent (non-functioning); both types frequently synthesize more than one peptide, although often these are not associated with specific syndromes. Based on data from various sources the incidence and prevalence of GI-NETs is increasing. Surgery is the only possible curative approach and so represents the traditional first-line therapy. However, as most patients with NETs are diagnosed once metastases have occurred, curative surgery is generally not possible. Patients therefore require medical management with the aim of relieving symptoms and suppressing tumor growth and spread. Somatostatin analogues can improve the symptoms of carcinoid syndrome and stabilize tumor growth (PROMID study) in many patients. An antiproliferative effect can also be achieved with the m-TOR inhibitor everolimus, alone or in combination with octreotide LAR. The vascular endothelial growth factor inhibitor sunitinib has demonstrated antitumor effects in pancreatic NETs. Pasireotide, the multi-receptor targeted somatostatin analogue, has the potential to be an effective therapy for de novo or octreotide-refractory carcinoid syndrome. Peptide receptor radiotherapy with yttrium 90-DOTATOC or lutetium 177-DOTATE are also new interesting treatment options for NETs.


The human AQP4 gene: definition of the locus encoding two water channel polypeptides in brain.
The aquaporin family of membrane water transport proteins are expressed in diverse tissues, and in brain the predominant water channel protein is AQP4. Here we report the isolation and characterization of the human AQP4 cDNAs and genomic DNA. Two cDNAs were isolated corresponding to the two initiating methionines (M1 in a 323-aa polypeptide and M23 in a 301-aa polypeptide) previously identified in rat [Jung, J.S., Bhat, R.V., Preston, G.M., Guggino, W.B. & Agre, P. (1994) Proc. Natl. Acad. Sci. USA 91, 13052-13056]. Similar to other aquaporins, the AQP4 gene is composed of four exons encoding 127, 55, 27, and 92 amino acids separated by introns of 0.8, 0.3, and 5.2 kb. Unlike other aquaporins, an alternative coding initiation sequence (designated exon 0) was located 2.7 kb upstream of exon 1. When spliced together, M1 and the subsequent 10 amino acids are encoded by exon 0; the next 11 amino acids and M23 are encoded by exon 1. Transcription initiation sites have been mapped in the proximal promoters of exons 0 and 1. RNase protection revealed distinct transcripts corresponding to M1 and M23 mRNAs, and AQP4 immunoblots of cerebellum demonstrated reactive polypeptides of 31 and 34 kDa. Using a P1 and a lambda EMBL subclone, the chromosomal site of the human AQP4 gene was mapped to chromosome 18 at the junction of q11.2 and q12.1 by fluorescence in situ hybridization. These studies may now permit molecular characterization of AQP4 during human development and in clinical disorders.


Effect of MeCP2 on TGF- β1-induced Extracellular Matrix Production in Nasal Polyp-derived Fibroblasts.
Purpose Methyl-CpG-binding protein 2 (MeCP2), known as a transcriptional regulator, has been suggested to play an important role in myofibroblast differentiation in the lung. The purpose of this study was to investigate the role of MeCP2 in transforming growth factor (TGF)- β1-induced myofibroblast differentiation and extracellular matrix (ECM) production in nasal polyp-derived fibroblasts (NPDFs). Methods To identify the expression of MeCP2 in nasal polyp tissues, immunohistochemistry staining and Western blot were performed. TGF- β1-induced NPDFs were treated with 5-azacytidine, a DNA methylation inhibitor, and the expression levels of α-SMA and fibronectin were determined by semiquantitative reverse transcription polymerase chain reaction, immunofluorescent staining, and Western blotting. The total soluble collagen was analyzed by the Sircol collagen assay. MeCP2 silenced by MeCP2-specific small interference ( si) RNA was verified by Western blot. Results The expression levels of MeCP2 increased in nasal polyp tissues compared to normal inferior turbinate tissues. 5-Azacytidine significantly inhibited the expression of α-SMA and fibronectin mRNA in a dose-dependent manner. In addition, 5-azacytidine suppressed collagen production and the expression of MeCP2 in the same manner. The expression levels of a-SMA and collagen production were significantly blocked by MeCP2 silencing in TGF- β1-induced NPDFs. Conclusions Our data suggest that MeCP2 plays an essential role in TGF- β1-induced myofibroblast differentiation and ECM production in NPDFs.


Etv6 activates vegfa expression through positive and negative transcriptional regulatory networks in Xenopus embryos.
VEGFA signaling controls physiological and pathological angiogenesis and hematopoiesis. Although many context-dependent signaling pathways downstream of VEGFA have been uncovered, vegfa transcriptional regulation in vivo remains unclear. Here, we show that the ETS transcription factor, Etv6, positively regulates vegfa expression during Xenopus blood stem cell development through multiple transcriptional inputs. In agreement with its established repressive functions, Etv6 directly inhibits expression of the repressor foxo3, to prevent Foxo3 from binding to and repressing the vegfa promoter. Etv6 also directly activates expression of the activator klf4; reflecting a genome-wide paucity in ETS-binding motifs in Etv6 genomic targets, Klf4 then recruits Etv6 to the vegfa promoter to activate its expression. These two mechanisms (double negative gate and feed-forward loop) are classic features of gene regulatory networks specifying cell fates. Thus, Etv6's dual function, as a transcriptional repressor and activator, controls a major signaling pathway involved in endothelial and blood development in vivo.


The quantitative effect of 0.5% ketorolac tromethamine solution and 0.1% dexamethasone sodium phosphate solution on postsurgical blood-aqueous barrier.
Anterior chamber fluorophotometry was performed after the oral administration of fluorescein sodium in patients undergoing extracapsular cataract extraction and posterior chamber intraocular lens insertion before and after surgery. The administration of 0.5% ketorolac tromethamine solution (ketorolac solution) eye drops before and after surgery decreased the breakdown of the blood-aqueous barrier as compared with 0.1% dexamethasone sodium phosphate solution (dexamethasone solution) eye drops at each period, as measured by fluorophotometry. A single injection below Tenon's capsule of a short-acting corticosteroid had been given to each patient at the end of each surgical procedure. Slit-lamp observations of postoperative ocular inflammation were not different between treatment groups. Both ketorolac and dexamethasone solutions were well tolerated by patients. Ketorolac solution was more effective than dexamethasone solution in facilitating reestablishment of the blood-aqueous barrier after surgery, as measured by fluorophotometry, and was equal to dexamethasone solution as observed by slit-lamp observations. This study suggests that ketorolac ophthalmic solution may be effective and safe as a nonsteroidal anti-inflammatory agent for topical use after cataract surgery and intraocular lens implantation in place of topically administered corticosteroids.


Reverse engineering the face space: Discovering the critical features for face identification.
How do we identify people? What are the critical facial features that define an identity and determine whether two faces belong to the same person or different people? To answer these questions, we applied the face space framework, according to which faces are represented as points in a multidimensional feature space, such that face space distances are correlated with perceptual similarities between faces. In particular, we developed a novel method that allowed us to reveal the critical dimensions (i.e., critical features) of the face space. To that end, we constructed a concrete face space, which included 20 facial features of natural face images, and asked human observers to evaluate feature values (e.g., how thick are the lips). Next, we systematically and quantitatively changed facial features, and measured the perceptual effects of these manipulations. We found that critical features were those for which participants have high perceptual sensitivity (PS) for detecting differences across identities (e.g., which of two faces has thicker lips). Furthermore, these high PS features vary minimally across different views of the same identity, suggesting high PS features support face recognition across different images of the same face. The methods described here set an infrastructure for discovering the critical features of other face categories not studied here (e.g., Asians, familiar) as well as other aspects of face processing, such as attractiveness or trait inferences.


[Effects of subtotal nephrectomy and high salt intake on atrial natriuretic peptide in the brains of rats].
In order to explore the role of brain atrial natriuretic peptide (ANP) in regulating water and electrolytes, the changes of the ANP content of various brain areas in the status of high salt intake and in 3/4 nephrectomy rats. In control group, a wide distribution of ANP was observed in different areas of the brain. The amount of daily water intake and urine were larger, and urine sodium concentration was lower in the 3/4 nephrectomy group than those in the control group (P < 0.05). In the ten nuclei we have studied, ANP content was not significantly different from that of the control (P > 0.05). In the high salt group, water intake, urine volume and urine sodium concentration were significantly higher than those in the control. In this group, brain ANP increased in the periventricular nuclues (PVN) and arcute nucleus (Arc) (P < 0.05). The results indicate that ANP in anteroventral region of the third ventricle (AV3V) might play important role in the regulation of water and electolytes.


Expression of VEGF and semaphorin genes define subgroups of triple negative breast cancer.
Triple negative breast cancers (TNBC) are difficult to treat due to a lack of targets and heterogeneity. Inhibition of angiogenesis is a promising therapeutic strategy, but has had limited effectiveness so far in breast cancer. To quantify heterogeneity in angiogenesis-related gene expression in breast cancer, we focused on two families--VEGFs and semaphorins--that compete for neuropilin co-receptors on endothelial cells. We compiled microarray data for over 2,600 patient tumor samples and analyzed the expression of VEGF- and semaphorin-related ligands and receptors. We used principal component analysis to identify patterns of gene expression, and clustering to group samples according to these patterns. We used available survival data to determine whether these clusters had prognostic as well as therapeutic relevance. TNBC was highly associated with dysregulation of VEGF- and semaphorin-related genes; in particular, it appeared that expression of both VEGF and semaphorin genes were altered in a pro-angiogenesis direction. A pattern of high VEGFA expression with low expression of secreted semaphorins was associated with 60% of triple-negative breast tumors. While all TNBC groups demonstrated poor prognosis, this signature also correlated with lower 5-year survival rates in non-TNBC samples. A second TNBC pattern, including high VEGFC expression, was also identified. These pro-angiogenesis signatures may identify cancers that are more susceptible to VEGF inhibition.


Functional implications of cross-orientation inhibition of cortical visual cells. I. Neurophysiological evidence.
Simple and complex cells of striate cortex of anaesthetized and paralysed cats were stimulated with two superimposed one-dimensional grating stimuli of different orientations to investigate inhibitory effects of non-optimally oriented stimuli. We confirmed that a stimulus of orientation orthogonal to a cell's long axis significantly reduces the cell's discharge rate. Further experiments revealed the following. (i) The inhibition was typically stronger for simple than for complex cells. (ii) It is very broadly tuned for orientation, all orientations outside the cell's tuning band having a comparable inhibitory effect. (iii) Similarly, it is broadly tuned for spatial frequency. These last two results suggest that the inhibition arises not from a single cell but from a pool of cells. (iv) The pattern of the discharge of the inhibition in response to stimulation by phase-reversed sinusoidal gratings is consistent with the notion that the inhibition arises from complex cells. A second series of recordings of stimulation by visual noise patterns demonstrated how 'cross-orientation inhibition' prevents simple cells from responding to two-dimensional visual noise while allowing them to respond to comparable one-dimensional noise patterns. We suggest that this mechanism may serve to render simple cells selectively sensitive to one-dimensional stimuli, such as the contours or borders of visual objects.


Evaluation of long-term patient satisfaction and experience with the Baha® bone conduction implant.
Evaluate long-term patient satisfaction with bone-anchored hearing aids (the Baha®, now referred to by Cochlear as a 'bone conduction implant') in our hospital clinic spanning the eighteen-year period from the inception of our Baha program. The researchers further wished to analyse the various factors leading to patient satisfaction/dissatisfaction with their Baha. We developed a new questionnaire to obtain a comprehensive impression of individual patient practices, general satisfaction, and experiences with their Baha in respect to time spent using Baha, sound quality, annoyance from noise disturbance, ease of communication, cosmetic appearance, and satisfaction with the Baha amongst patient relatives, an aspect not previously investigated. The study design was retrospective and executed as a postal questionnaire. The questionnaire was developed by the authors of this paper. Patients operated on for a Baha at our hospital from 1989 to 2007. The response rate was 92.4%. Eighty-six percent were satisfied or very satisfied with their Baha. Ninety-one percent of respondents could communicate using their Baha in a one-on-one conversational setting. A primary factor leading to dissatisfaction, experienced by 70% of responding patients, was annoyance from wind noise. Baha was found to yield good overall patient satisfaction over the long-term, and it was possible to identify specific factors attributing to satisfaction/dissatisfaction.


Multiple brain metastases - current management and perspectives for treatment with electrochemotherapy.
Due to the advanced oncological treatments of cancer, an overall increase in cancer incidence, and better diagnostic tools, the incidence of brain metastases is on the rise. This review addresses the current treatment options for patients with multiple brain metastases, presenting electrochemotherapy (ECT) as one of the new experimental treatments for this group of patients. Neurosurgery, stereotactic surgery, and whole-brain radiotherapy are the evidence-based treatments that can be applied for patients with multiple brain metastases. Treatment with chemotherapy and molecularly targeted agents may also be warranted. Several experimental treatments are emerging, one of which is ECT, an effective cancer treatment comprising electric pulses given by electrodes in the tumor tissue, causing electroporation of the cell membrane, and thereby augmenting uptake and the cytotoxicity of the chemotherapeutic drug bleomycin by 300 times. Preclinical data are promising and the first patient has been treated in an ongoing clinical trial for patients with brain metastases. Perspectives for ECT in the brain include treatment of primary and secondary brain tumors as well as soft tissue metastases elsewhere.


Molecular interactions of serotonin (5-HT) and endothelin-1 in vascular smooth muscle cells: in vitro and ex vivo analyses.
Elevated levels of serotonin (5-HT) and endothelin-1 (ET-1) may be involved in cardiovascular complications of diabetes mellitus. Data suggest supraphysiological concentrations of 5-HT (10(-6) M) potentiate the ability of ET-1 to stimulate DNA synthesis and vascular smooth muscle cell (VSMC) proliferation in vitro via activation of mitogen-activated protein kinase (p42/44 MAPK) and Janus kinase 2 (JAK2) pathways. Additionally, 5-HT enhances agonist-induced contractions via p42/44 MAPK and an unknown tyrosine kinase. However, the exact mechanisms of the 5-HT/ET-1 interactions and whether these effects occur at physiological levels (10(-9) M) are unknown. Therefore, we hypothesized that interactions between 5-HT and ET-1 at physiological concentrations in VSMC enhanced activation of both p42/44 MAPK and JAK2 pathways contributing to vascular growth and contractile responses. With the use of rat VSMC and Western blot analysis, our data suggest no effect of acute (30 min) preincubation with 5-HT (10(-9) M) and/or ET-1 (10(-9) M) on the activation of either pathway in normal or high glucose conditions. To determine if there was altered vascular reactivity in intact vessels we tested the effects of 5-HT and ET-1 interaction using myographs to measure isometric contractions of rat thoracic aortic rings. 5-HT (10(-9) M) and ET-1 (10(-12) M) stimulate enhanced contractile responses to each other that were inhibited by JAK2 and p42/44 MAPK antagonists. Our findings demonstrate that both 5-HT and ET-1 at physiological concentrations could interact with each other and activate p42/44 MAPK and JAK2 signaling pathways to cause an increase in smooth muscle contraction that could lead to altered vascular function.


Molecular mechanisms of muscular dystrophies: old and new players.
The study of the muscle cell in the muscular dystrophies (MDs) has shown that mutant proteins result in perturbations of many cellular components. MDs have been associated with mutations in structural proteins, signalling molecules and enzymes as well as mutations that result in aberrant processing of mRNA or alterations in post-translational modifications of proteins. These findings have not only revealed important insights for cell biologists, but have also provided unexpected and exciting new approaches for therapy.


The representation of oral fat texture in the human somatosensory cortex.
How fat is sensed in the mouth and represented in the brain is important in relation to the pleasantness of food, appetite control, and the design of foods that reproduce the mouthfeel of fat yet have low energy content. We show that the human somatosensory cortex (SSC) is involved in oral fat processing via functional coupling to the orbitofrontal cortex (OFC), where the pleasantness of fat texture is represented. Using functional MRI, we found that activity in SSC was more strongly correlated with the OFC during the consumption of a high fat food with a pleasant (vanilla) flavor compared to a low fat food with the same flavor. This effect was not found in control analyses using high fat foods with a less pleasant flavor or pleasant-flavored low fat foods. SSC activity correlated with subjective ratings of fattiness, but not of texture pleasantness or flavor pleasantness, indicating a representation that is not involved in hedonic processing per se. Across subjects, the magnitude of OFC-SSC coupling explained inter-individual variation in texture pleasantness evaluations. These findings extend known SSC functions to a specific role in the processing of pleasant-flavored oral fat, and identify a neural mechanism potentially important in appetite, overeating, and obesity.


Human milk, breastfeeding, and transmission of human immunodeficiency virus type 1 in the United States. American Academy of Pediatrics Committee on Pediatric AIDS.
Transmission of human immunodeficiency virus type 1 (HIV-1) through breastfeeding has been conclusively demonstrated. The risk of such transmission has been quantified, the timing has been clarified, and certain risk factors for breastfeeding transmission have been identified. In areas where infant formula is accessible, affordable, safe, and sustainable, avoidance of breastfeeding has represented one of the main components of mother-to-child HIV-1 transmission prevention efforts for many years. In areas where affordable and safe alternatives to breastfeeding may not be available, interventions to prevent breastfeeding transmission are being investigated. Complete avoidance of breastfeeding by HIV-1-infected women has been recommended by the American Academy of Pediatrics and the Centers for Disease Control and Prevention and remains the only means by which prevention of breastfeeding transmission of HIV-1 can be absolutely ensured. This technical report summarizes the information available regarding breastfeeding transmission of HIV-1.


Renal cell carcinoma: biological features and rationale for molecular-targeted therapy.
Cancer cells are characterized by indefinite proliferation, invasiveness and metastases. These characteristics are usually related to one another. Namely, cancer cells that proliferate rapidly tend to invade and metastasize. Renal cell carcinoma (RCC) typically does not proliferate rapidly nor does it invade the surrounding tissues, but it does metastasize. RCC has several peculiar characteristics that are not observed in other cancers: a relatively late recurrence, a high frequency of paraneoplastic syndrome, hypervascularity and the spontaneous regression of metastatic lesions after the excision of the primary tumor. These clinical observations suggest that cytokines or growth factors are important contributors to microenvironments favoring the growth of cancer cells. Thus, the blockade of cell-to-cell communication might have some therapeutic potential. Accordingly, a popular strategy for molecular-targeted therapy for RCC targets the vasculization of RCC induced by vascular endothelial growth factor (VEGF). This review highlights the biological features of RCC that are relevant to molecular-targeted therapy.


Shear force at failure and stiffness of all-inside meniscal repair devices.
The purpose of this study was to determine the failure load and stiffness of various meniscal repair devices. A total of 61 fresh-frozen porcine menisci (medial and lateral) were used for the study. A 30-mm vertical, full-thickness tear was created and repaired using one of three all-inside fixation devices and one inside-out repair in the vertical mattress pattern. We used the MaxBraid (Biomet, Warsaw, IN) inside-out suture as a control. The other devices tested were the Meniscal Cinch (Arthrex, Naples, FL), Ultra FasT-Fix (Smith & Nephew, Andover, MA), and the MaxFire MarXmen (Biomet, Warsaw, IN). In addition, two devices, MaxFire MarXmen and Ultra FasT-Fix, were tested using a horizontal mattress configuration. Using the vertical mattress pattern, the Meniscal Cinch had the highest average load to failure. The Meniscal Cinch was significantly less stiff than the other three devices (p < 0.04). For the MarXmen and Ultra FasT-Fix, no differences were noted for load to failure between horizontal and vertical mattress patterns. The mode of failure was significantly different when comparing the two different surgical techniques for the MaxFire MarXmen (p = 0.005). The MaxFire MarXmen device produced a significantly stiffer (p < 0.001) construct when following the manufacturer's instructions (5.8 N/mm) than with the technique used for the other all-inside devices (2.5 N/mm) The Meniscal Cinch had the highest load-to-failure value but the lowest stiffness of the group in the vertical mattress configuration. There was little difference in biomechanical properties between vertical and horizontal repair. Importantly, there was a significant difference in stiffness and failure mode for the MaxFire MarXmen when the manufacturer guidelines were not specifically followed.


Youth drinking in decline: What are the implications for public health, public policy and public debate?
Youth drinking has declined across most high-income countries in the last 20 years. Although researchers and commentators have explored the nature and drivers of decline, they have paid less attention to its implications. This matters because of the potential impact on contemporary and future public health, as well as on alcohol policy-making. This commentary therefore considers how youth drinking trends may develop in future, what this would mean for public health, and what it might mean for alcohol policy and debate. We argue that the decline in youth drinking is well-established and unlikely to reverse, despite smaller declines and stabilising trends in recent years. Young people also appear to be carrying their lighter drinking into adulthood in at least some countries. This suggests we should expect large short- and long-term public health benefits. The latter may however be obscured in population-level data by increased harm arising from earlier, heavier drinking generations moving through the highest risk points in the life course. The likely impact of the decline in youth drinking on public and policy debate is less clear. We explore the possibilities using two model scenarios, the reinforcement and withdrawal models. In the reinforcement model, a 'virtuous' circle of falling alcohol consumption, increasing public support for alcohol control policies and apparent policy successes facilitates progressive strengthening of policy, akin to that seen in the tobacco experience. In the withdrawal model, policy-makers turn their attention to other problems, public health advocates struggle to justify proposed interventions and existing policies erode over time as industry actors reassert and strengthen their partnerships with government around alcohol policy. We argue that disconnects between the tobacco experience and the reinforcement model make the withdrawal model a more plausible scenario. We conclude by suggesting some tentative ways forward for public health actors working in this space.


Developing, Implementing, and Evaluating a Multimedia Patient Decision Aid Program to Reform the Informed Consent Process of a Peripherally Inserted Central Venous Catheter Procedure: Protocol for Quality Improvement.
Informed consent has considerable clinical, ethical, and legal implications for patient safety and liability. Little information is available about the use of multimedia patient decision aids (PtDA) in the consent process for therapeutic invasive procedures such as the peripherally inserted central venous catheter (PICC). In addition, none of the available studies have designed their multimedia PtDAs based on the Agency for Healthcare Research and Quality's (AHRQ) comprehensive guide for informed consent. This paper describes a patient-centered, systematic, multidisciplinary approach to develop, implement, and alpha test a multimedia PtDA to reform the informed consent process of a PICC for patients in 10 acute and intensive care units. The development, implementation, and evaluation processes of the PtDA followed the phases in the Multimedia Production Framework: preproduction, production, and postproduction. Within this framework, we applied the criteria for judging the quality of PtDAs, the AHRQ's Health Literacy Universal Precautions Toolkit, and the AHRQ's Patient Education Materials Assessment Tool Guide. The methodology was guided by the Interprofessional Shared Decision-Making Model and the AHRQ's Making Informed Consent an Informed Choice guide. In the preproduction phase, we (1) reviewed the current consent form; (2) observed 18 consent processes; (3) surveyed the vascular access team (N=6 nurses) about their perception of the current process; (4) surveyed 30 patients for knowledge recall and retention, overall satisfaction, and attitude toward using a multimedia PtDA; and (5) wrote and reviewed the script for the multimedia program. The production phase focused on filming the PtDA in English and Spanish languages. The postproduction phase included integrating the multimedia programs into the care processes, developing a modified workflow for the consent process, and alpha testing of the English and Spanish PtDAs by (1) a group of 5 patients for clarity and understandability of the information; (2) nurses using the AHRQ's Patient Education Materials Assessment Tool Audio and Video; and (3) by the multidisciplinary change team. Based on the alpha testing, patients indicated that the content was easy to follow and read; nurses provided positive feedback, and their comments were mainly related to the changes in the workflow in the consent process of the PICC after using the PtDA; and the multidisciplinary change team suggested edits related to changing a few scenes. The final multimedia program consisted of 7 min and 37 s demonstrating detailed information about the PICC. A systematic development of PtDAs for nonurgent invasive procedures may eliminate many limitations of the conventional consent process by ensuring comprehensive, standardized, and easy-to-comprehend information and providing sufficient time for the patients to reflect on the information. To be effective, PtDAs should follow a systematic, patient-centered, evidence-based, and rigorous approach in the development, implementation, and evaluation processes. RR1-10.2196/10709.


Ubiquitin Specific Protease 26 (USP26) expression analysis in human testicular and extragonadal tissues indicates diverse action of USP26 in cell differentiation and tumorigenesis.
Ubiquitin specific protease 26 (USP26), a deubiquitinating enzyme, is highly expressed early during murine spermatogenesis, in round spermatids, and at the blood-testis barrier. USP26 has also been recognized as a regulator of androgen receptor (AR) hormone-induced action involved in spermatogenesis and steroid production in in vitro studies. Prior mutation screening of USP26 demonstrated an association with human male infertility and low testosterone production, but protein localization and expression in the human testis has not been characterized previously. USP26 expression analysis of mRNA and protein was completed using murine and human testis tissue and human tissue arrays. USP26 and AR mRNA levels in human testis were quantitated using multiplex qRT-PCR. Immunofluorescence colocalization studies were performed with formalin-fixed/paraffin-embedded and frozen tissues using primary and secondary antibodies to detect USP26 and AR protein expression. Human microarray dot blots were used to identify protein expression in extra-gonadal tissues. For the first time, expression of USP26 and colocalization of USP26 with androgen receptor in human testis has been confirmed predominantly in Leydig cell nuclei, with less in Leydig cell cytoplasm, spermatogonia, primary spermatocytes, round spermatids, and Sertoli cells. USP26 likely affects regulatory proteins of early spermatogenesis, including androgen receptor with additional activity in round spermatids. This X-linked gene is not testis-specific, with USP26 mRNA and protein expression identified in multiple other human organ tissues (benign and malignant) including androgen-dependent tissues such as breast (myoepithelial cells and secretory luminal cells) and thyroid tissue (follicular cells). USP26/AR expression and interaction in spermatogenesis and androgen-dependent cancer warrants additional study and may prove useful in diagnosis and management of male infertility.


Isoenzymic characterization of seven strains of Toxoplasma gondii by isoelectrofocusing in polyacrylamide gels.
Toxoplasma gondii strains are usually defined by biological parameters such as pathogenicity in mice. A characterization of toxoplasma strains by biochemical techniques has not been reported. In this study, extracts of tachyzoites of 7 toxoplasma strains were compared on the basis of their isoenzyme patterns for 39 enzymes by means of isoelectrofocusing in polyacrylamide gels. Eighteen enzymes gave clear and reproducible bands. Of these, 14 had identical electrophoretic patterns for all strains. Two different isoenzyme types were found for the enzymes aspartate aminotransferase, glutathione reductase, glucose phosphate isomerase, and amylase. This allowed the description of 3 isoenzyme pattern groups among the 7 toxoplasma strains. The possible relationship between biological behavior and isoenzyme pattern groups is discussed.


Force and twist dependence of RepC nicking activity on torsionally-constrained DNA molecules.
Many bacterial plasmids replicate by an asymmetric rolling-circle mechanism that requires sequence-specific recognition for initiation, nicking of one of the template DNA strands and unwinding of the duplex prior to subsequent leading strand DNA synthesis. Nicking is performed by a replication-initiation protein (Rep) that directly binds to the plasmid double-stranded origin and remains covalently bound to its substrate 5'-end via a phosphotyrosine linkage. It has been proposed that the inverted DNA sequences at the nick site form a cruciform structure that facilitates DNA cleavage. However, the role of Rep proteins in the formation of this cruciform and the implication for its nicking and religation functions is unclear. Here, we have used magnetic tweezers to directly measure the DNA nicking and religation activities of RepC, the replication initiator protein of plasmid pT181, in plasmid sized and torsionally-constrained linear DNA molecules. Nicking by RepC occurred only in negatively supercoiled DNA and was force- and twist-dependent. Comparison with a type IB topoisomerase in similar experiments highlighted a relatively inefficient religation activity of RepC. Based on the structural modeling of RepC and on our experimental evidence, we propose a model where RepC nicking activity is passive and dependent upon the supercoiling degree of the DNA substrate.


A molecular imprinted SPR biosensor for sensitive determination of citrinin in red yeast rice.
A novel and sensitive molecular imprinted surface plasmon resonance (SPR) biosensor was developed for selective determination of citrinin (CIT) in red yeast rice. Firstly, the gold surface of SPR chip was modified with allyl mercaptane. Then, CIT-imprinted poly(2-hydroxyethyl methacrylate-methacryloylamidoglutamic acid) (p(HEMA-MAGA)) film was generated on the gold surface modified with allyl mercaptane. The unmodified and imprinted surfaces were characterized by Fourier transform infrared (FTIR) spectroscopy, atomic force microscopy (AFM) and contact angle measurements. The linearity range and the detection limit were obtained as 0.005-1.0 ng/mL and 0.0017 ng/mL, respectively. The SPR biosensor was applied to determination of CIT in red yeast rice sample.


Acceptance of provider-initiated testing and counseling for HIV infection by caregivers in a tertiary health institution in Abuja, Nigeria: a cross sectional study.
Less than 10% of HIV positive children are enrolled into antiretroviral treatment program in the country. Provider-initiated testing and counseling was introduced to increasing uptake of HIV testing. The aim of this study is to determine the acceptability and factors undermining the acceptance of this laudable initiative by parents/caregivers of children attending paediatric out patient clinical services in our health institution. A cross sectional study of children aged 18 months to 18 years and their parents/caregivers attending paediatric outpatient clinic of the hospital was undertaken for the above objectives. There were statistically more female parents/caregivers (82.5%, p=0.00), more male patients (52.9 %, p= 0.02), and 11.9% adolescents in this study. While 91.7% of parents/caregivers admitted not having knowledge of provider-initiated testing and counseling, 95.6% knew what HIV was. Acceptance of the program was high (98.7%), majority (89.7%) wanting to know the HIV status of their children/wards. Non-acceptance was small (1.2%), there main reason being prior knowledge of their HIV status. Prevalence of HIV among tested children was 1.7%. There was a strong relationship between having willingness to test for HIV and many of the study variables with religion of the parents/caregivers having the strongest relationship [OR: 13.94, (CI 1.82, 55.34)], and tribe having list association, [OR: 3.60, (CI 1.85, 17.14)]. There was general wiliness to accept HIV test for children by their parents/caregiver in this study, and HIV prevalence in children is on a downward trend; its sustenance to be continued and adolescent clinics need to be created.


Recognising the style of spatially exaggerated tennis serves.
A technique for the construction of exaggerated human movements was developed and its effectiveness tested for the case of categorising tennis serves as flat, slice, or topspin. The technique involves treating movements as points in a high-dimensional space and uses average movements as the basis for constructing exaggerated movements. Exaggerated movements of a particular style are defined as those points in the space of movements which lie on a line originating at the style average and in the direction defined by the difference between the style average and the grand average. In order to visualise the movements, computer animation techniques were employed to transform the three-dimensional coordinates of the movement into the motion of a solid-body figure. These solid-body models were used in perceptual experiments to assess the effectiveness of the exaggeration technique. After an initial training session on the exemplars from the original library, subjects viewed the synthetic tennis-serve motions and in two separate sessions either made three-alternative, categorisation judgments after viewing a single serve or rated dissimilarity after viewing a pair of serves. Results from both accuracy in the categorisation task and structure of a multidimensional scaling solution of the matrix of dissimilarities indicated that, as distance from the grand average increased, the service motion became more distinct and more accurately identified.


Population growth dynamics of carbon nanotubes.
Understanding the population growth behavior of filamentary nanostructures, such as carbon nanotubes (CNTs), is hampered by the lack of characterization techniques capable of probing statistical variations with high spatial resolution. We present a comprehensive methodology for studying the population growth dynamics of vertically aligned CNT forests, utilizing high-resolution spatial mapping of synchrotron X-ray scattering and attenuation, along with real-time height kinetics. We map the CNT alignment and dimensions within CNT forests, revealing broadening and focusing of size distributions during different stages of the process. Then, we calculate the number density and mass density of the CNT population versus time, which are true measures of the reaction kinetics. We find that the mass-based kinetics of a CNT population is accurately represented by the S-shaped Gompertz model of population growth, although the forest height and CNT length kinetics are essentially linear. Competition between catalyst activation and deactivation govern the rapid initial acceleration and slow decay of the CNT number density. The maximum CNT density (i.e., the overall catalyst activity) is limited by gas-phase reactions and catalyst-surface interactions, which collectively exhibit autocatalytic behavior. Thus, we propose a comprehensive picture of CNT population growth which combines both chemical and mechanical cooperation. Our findings are relevant to both bulk and substrate-based CNT synthesis methods and provide general insights into the self-assembly and collective growth of filamentary nanostructures.


Volunteers in plastic surgery guidelines for providing surgical care for children in the less developed world: part II. Ethical considerations.
Many international volunteer groups provide free reconstructive plastic surgery for the poor and underserved in developing countries. An essential issue in providing this care is that it meets consistent guidelines for both quality and safety-a topic that has been addressed previously. An equally important consideration is how to provide that care in an ethical manner. No literature presently addresses the various issues involved in making those decisions. With these ethical considerations in mind, the Volunteers in Plastic Surgery Committee of the American Society of Plastic Surgeons/Plastic Surgery Foundation undertook a project to create a comprehensive set of guidelines for volunteer groups planning to provide this type of reconstructive plastic surgery in developing countries. The committee worked in conjunction with the Society for Pediatric Anesthesia on this project. The Board of the American Society of Plastic Surgeons/Plastic Surgery Foundation has approved the ethical guidelines created for the delivery of care in developing countries. The guidelines address the variety of ethical decisions that may be faced by a team working in an underdeveloped country. These guidelines make it possible for a humanitarian effort to anticipate the types of ethical decisions that are often encountered and be prepared to deal with them appropriately. Any group seeking to undertake an international mission trip in plastic surgery should be able to go to one source to find a detailed discussion of the perceived needs in providing ethical humanitarian care. This document was created to satisfy that need and is a companion to our original guidelines addressing safety and quality.


Caffeine increases motor output entropy and performance in 4 km cycling time trial.
Caffeine improves cycling time trial performance through enhanced motor output and muscle recruitment. However, it is unknown if caffeine further increases power output entropy. To investigate the effects of caffeine effects on cycling time trial performance and motor output entropy (MOEn), nine cyclists (VO2MAX of 55 ± 6.1 mL.kg.-1min-1) performed a 4 km cycling time trial (TT4km) after caffeine and placebo ingestion in a counterbalanced order. Power output data were sampled at a 2 Hz frequency, thereafter entropy was estimated on a sliding-window fashion to generate a power output time series. A number of mixed models compared performance and motor output entropy between caffeine and placebo every 25% of the total TT4km distance. Caffeine ingestion improved power output by 8% (p = 0.003) and increased MOEn by 7% (p = 0.018). Cyclists adopted a U-shaped pacing strategy after caffeine ingestion. MOEn mirrored power output responses as an inverted U-shape MOEn during the time trial. Accordingly, a strong inverse correlation was observed between MOEn and power output responses over the last 25% of the TT4km (p < 0.001), regardless of the ingestion, likely reflecting the end spurt during this period (p = 0.016). Caffeine ingestion improved TT4km performance and motor output responses likely due to a greater power output entropy.


Pre- and early-postnatal nutrition modify gene and protein expressions of muscle energy metabolism markers and phospholipid Fatty Acid composition in a muscle type specific manner in sheep.
We previously reported that undernutrition in late fetal life reduced whole-body insulin sensitivity in adult sheep, irrespective of dietary exposure in early postnatal life. Skeletal muscle may play an important role in control of insulin action. We therefore studied a range of putative key muscle determinants of insulin signalling in two types of skeletal muscles (longissimus dorsi (LD) and biceps femoris (BF)) and in the cardiac muscle (ventriculus sinister cordis (VSC)) of sheep from the same experiment. Twin-bearing ewes were fed either 100% (NORM) or 50% (LOW) of their energy and protein requirements during the last trimester of gestation. From day-3 postpartum to 6-months of age (around puberty), twin offspring received a high-carbohydrate-high-fat (HCHF) or a moderate-conventional (CONV) diet, whereafter all males were slaughtered. Females were subsequently raised on a moderate diet and slaughtered at 2-years of age (young adults). The only long-term consequences of fetal undernutrition observed in adult offspring were lower expressions of the insulin responsive glucose transporter 4 (GLUT4) protein and peroxisome proliferator-activated receptor gamma, coactivator 1α (PGC1α) mRNA in BF, but increased PGC1α expression in VSC. Interestingly, the HCHF diet in early postnatal life was associated with somewhat paradoxically increased expressions in LD of a range of genes (but not proteins) related to glucose uptake, insulin signalling and fatty acid oxidation. Except for fatty acid oxidation genes, these changes persisted into adulthood. No persistent expression changes were observed in BF and VSC. The HCHF diet increased phospholipid ratios of n-6/n-3 polyunsaturated fatty acids in all muscles, even in adults fed identical diets for 1½ years. In conclusion, early postnatal, but not late gestation, nutrition had long-term consequences for a number of determinants of insulin action and metabolism in LD. Tissues other than muscle may account for reduced whole body insulin sensitivity in adult LOW sheep.


Demonstration of the anti-viral activity of garlic extract against human cytomegalovirus in vitro.
The in vitro anti-viral activity of garlic extract (GE) on human cytomegalovirus (HCMV) was evaluated by tissue culture, plaque reduction and early antigen assay. A dose dependent inhibitory effect of GE was evident when GE was applied simultaneously with HCMV. But the effect was stronger when the monolayers were pretreated with GE. In addition, the anti-viral effect of GE persisted long in infected cells after its being removed from the culture medium. The strongest anti-viral effect of GE was demonstrated when it was applied continuously. It is therefore recommended that clinical use of GE against HCMV infection should be persistent and the prophylactic use of GE is preferable in immunocompromised patients.


'We are despised in the hospitals': sex workers' experiences of accessing health care in four African countries.
Sex workers in east and southern Africa are exposed to multiple occupational health and safety risks. Detailed understanding of barriers to accessing care would optimise design of improved services for this population. In this study, trained sex workers conducted 55 in-depth interviews and 12 focus group discussions with 106 female, 26 male and 4 transgender sex workers across 6 urban sites in Kenya, Zimbabwe, Uganda and South Africa. Data were analysed thematically, following an interpretive framework. Participants cited numerous unmet health needs, including diagnosis and treatment for sexually transmitted infections and insufficient access to condoms and lubricant. Denial of treatment for injuries following physical assault or rape and general hostility from public-sector providers was common. Resources permitting, many sex workers attended private services, citing higher quality and respect for dignity and confidentiality. Sex workers in southern Africa accessed specialised sex worker clinics, reporting mostly positive experiences. Across sites, participants called for additional targeted services, but also sensitisation and training of public-sector providers. Criminalisation of sex workers and associated stigmatisation, particularly of transgender and male sex workers, hinder HIV-prevention efforts and render access to mainstream healthcare precarious. Alongside law reform, sex worker-led peer outreach work should be strengthened and calls by sex workers for additional targeted services heeded.


Relation of hormone-replacement therapy to measures of plasma fibrinolytic activity. Atherosclerosis Risk in Communities (ARIC) Study Investigators.
The mechanisms by which replacement hormones may reduce the risk of coronary heart disease are not fully understood. Of specific interest is a potential effect of replacement hormones on plasma fibrinolytic activity, a putative determinant of thrombotic events. We investigated the relation of current use of replacement hormones to three measures of plasma fibrinolytic activity: tissue-type plasminogen activator (TPA) antigen, plasminogen activator inhibitor-1 (PAI-1) antigen, and D-dimer. The sample was composed of 288 women, free of clinical cardiovascular disease, who were selected for a case-control study of atherosclerosis: 142 women with ultrasonographic evidence of carotid intimal-medial thickening (cases) and 146 control subjects. Twenty percent (59 women) reported current use of replacement hormones. TPA antigen and PAI-1 antigen were highly correlated with each other (r = .67), whereas D-dimer correlated only weakly with TPA or PAI-1. Compared with nonusers, current users of replacement hormones had lower mean levels of TPA and PAI-1 antigens, suggesting enhanced fibrinolytic potential. In the entire sample, the multivariate-adjusted geometric mean values of TPA antigen were 6.3 and 7.3 ng/mL among current users and nonusers, respectively (P = .01); the corresponding values for PAI-1 antigen were 6.1 and 7.5 ng/mL (P = .13). These results were generally consistent for both atherosclerosis cases and their control subjects. D-dimer levels were lower in current hormone users than in nonusers, but the difference was not statistically significant (P > .15) in any of the analyses. The use of replacement hormones appears to be associated with enhancement of endogenous fibrinolytic potential. Enhanced plasma fibrinolytic activity among hormone users may explain, in part, the inverse association between hormone replacement therapy and coronary heart disease.


Mock jurors' evaluation of firearm examiner testimony.
Firearms experts traditionally have testified that a weapon leaves "unique" toolmarks, so bullets or cartridge casings can be visually examined and conclusively matched to a particular firearm. Recently, due to scientific critiques, Department of Justice policy, and judges' rulings, firearms experts have tempered their conclusions. In two experiments, we tested whether this ostensibly more cautious language has its intended effect on jurors (Experiment 1), and whether cross-examination impacts jurors' perception of firearm testimony (Experiment 2). Four hypotheses were tested. First, jurors will accord significant weight to firearm testimony that declares a "match" compared to testimony that does not (Experiments 1 and 2). Second, variations to "match" language will not affect guilty verdicts (Experiment 1). Third, only the most cautious language ("cannot exclude the gun") would lower guilty verdicts (Experiment 1). Fourth, cross-examination will reduce guilty verdicts depending on specific language used (Experiment 2). In two preregistered, high-powered experiments with 200 mock jurors per cell, participants recruited from Qualtrics Panels were presented with a criminal case containing firearms evidence, which varied the wording of the examiner's conclusion and whether cross-examination was present. These variations include conclusion language used by practitioners, language advised by government organizations, and language required by judges in several cases. Participants gave a verdict, rated the evidence and expert in all conditions. Guilty verdicts significantly increased when a match was declared compared to when a match was not declared. Variation in conclusion language did not affect guilty verdicts nor did it affect jurors' estimates of the likelihood the defendant's gun fired the bullet recovered at the crime scene. In contrast, however, a more cautious conclusion that an examiner "cannot exclude the defendant's gun" did significantly reduce guilty verdicts and likelihood estimates alike. The presence of cross-examination did not affect these findings. Apart from the most limited language ("cannot exclude the defendant's gun"), judicial intervention to limit firearms conclusion language is not likely to produce its intended effect. Moreover, cross-examination does not appear to affect perceptions or individual juror verdicts. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Effects of the agility boot camp with cognitive challenge (ABC-C) exercise program for Parkinson's disease.
Few exercise interventions practice both gait and balance tasks with cognitive tasks to improve functional mobility in people with PD. We aimed to investigate whether the Agility Boot Camp with Cognitive Challenge (ABC-C), that simultaneously targets both mobility and cognitive function, improves dynamic balance and dual-task gait in individuals with Parkinson's disease (PD). We used a cross-over, single-blind, randomized controlled trial to determine efficacy of the exercise intervention. Eighty-six people with idiopathic PD were randomized into either an exercise (ABC-C)-first or an active, placebo, education-first intervention and then crossed over to the other intervention. Both interventions were carried out in small groups led by a certified exercise trainer (90-min sessions, 3 times a week, for 6 weeks). Outcome measures were assessed Off levodopa at baseline and after the first and second interventions. A linear mixed-effects model tested the treatment effects on the Mini-BESTest for balance, dual-task cost on gait speed, SCOPA-COG, the UPDRS Parts II and III and the PDQ-39. Although no significant treatment effects were observed for the Mini-BESTest, SCOPA-COG or MDS-UPDRS Part III, the ABC-C intervention significantly improved the following outcomes: anticipatory postural adjustment sub-score of the Mini-BESTest (p = 0.004), dual-task cost on gait speed (p = 0.001), MDS-UPDRS Part II score (p = 0.01), PIGD sub-score of MDS-UPDRS Part III (p = 0.02), and the activities of daily living domain of the PDQ-39 (p = 0.003). Participants with more severe motor impairment or more severe cognitive dysfunction improved their total Mini-BESTest scores after exercise. The ABC-C exercise intervention can improve specific balance deficits, cognitive-gait interference, and perceived functional independence and quality of life, especially in participants with more severe PD, but a longer period of intervention may be required to improve global cognitive and motor function.


Robotic Pyelolithotomy, Extended Pyelolithotomy, Nephrolithotomy, and Anatrophic Nephrolithotomy.
We are a reporting on the indications, techniques, and limitations of robotic surgery in the management of renal stones disease. Robotic surgery is a good tool to manage large kidney and ureteral stones, particularly in patients with anatomic anomalies. We describe three techniques in managing staghorn kidney stones: robotic anatrophic nephrolithotomy, robotic pyelolithotomy, and robotic nephrolithotomy. Robotic pyelolithotomy (RP) is ideal for patients with large renal pelvis and partial staghorn stone with a wide extra-renal pelvis. Robotic nephrolithotomy (RN) is ideal for stones inside a calyceal diverticulum or a partial staghorn eroding into the renal parenchyma. Renal vascular control could be avoided in most of those surgeries. Robotic anatrophic nephrolithotomy (RAN) is the most complex procedure and is reserved for patients with complete staghorn stones when percutaneous approach was not successful or not feasible. Control of renal vasculature is required for RAN. For robotic kidney surgeries, patients are positioned in a lateral decubitus position. Four or five ports are placed based on the stone location and surgeon's preference. We prefer the trans-peritoneal approach as it gives us the optimal exposure. For RP and RN, hilar control is usually not required. The renal pelvis/ renal parenchyma is incised, and the stones are carefully removed. If needed intra-operative flexible nephoscopy can be used to remove residual stones fragments. The collecting system is closed using an absorbable suture. DJ stent if needed is placed in an antegrade fashion. For RAN, the kidney is fully mobilized, and hilar control is required to avoid excessive bleeding. The kidney is incised along Brodel's line and the stones are extracted. The kidney parenchyma is then closed using 1 or 2 layers. We achieved an almost 100% stone free rate with RP and RN. RAN remains a challenging procedure with a success rate around 70%. Robotic surgery is a viable option to manage large renal and ureteral stones particularly in situations where endoscopic approach is not successful or feasible.


The melatonin rhythm-generating enzyme: molecular regulation of serotonin N-acetyltransferase in the pineal gland.
A remarkably constant feature of vertebrate physiology is a daily rhythm of melatonin in the circulation, which serves as the hormonal signal of the daily light/dark cycle: melatonin levels are always elevated at night. The biochemical basis of this hormonal rhythm is one of the enzymes involved in melatonin synthesis in the pineal gland-the melatonin rhythm-generating enzyme-serotonin N-acetyltransferase (arylalkylamine N-acetyltransferase, AA-NAT, E.C. 22.214.171.124). In all vertebrates, enzyme activity is high at night. This reflects the influences of internal circadian clocks and of light. The dynamics of this enzyme are remarkable. The magnitude of the nocturnal increase in enzyme activity ranges from 7- to 150-fold on a species-to-species basis among vertebrates. In all cases the nocturnal levels of AA-NAT activity decrease very rapidly following exposure to light. A major advance in the study of the molecular basis of these changes was the cloning of cDNA encoding the enzyme. This has resulted in rapid progress in our understanding of the biology and structure of AA-NAT and how it is regulated. Several constant features of this enzyme have become apparent, including structural features, tissue distribution, and a close association of enzyme activity and protein. However, some remarkable differences among species in the molecular mechanisms involved in regulating the enzyme have been discovered. In sheep, AA-NAT mRNA levels show relatively little change over a 24-hour period and changes in AA-NAT activity are primarily regulated at the protein level. In the rat, AA-NAT is also regulated at a protein level; however, in addition, AA-NAT mRNA levels exhibit a 150-fold rhythm, which reflects cyclic AMP-dependent regulation of expression of the AA-NAT gene. In the chicken, cyclic AMP acts primarily at the protein level and a rhythm in AA-NAT mRNA is driven by a noncyclic AMP-dependent mechanism linked to the clock within the pineal gland. Finally, in the trout, AA-NAT mRNA levels show little change and activity is regulated by light acting directly on the pineal gland. The variety of mechanisms that have evolved among vertebrates to achieve the same goal-a rhythm in melatonin-underlines the important role melatonin plays as the hormonal signal of environmental lighting in vertebrates.


HLA class II antigens (DR, DQ loci) and peripheral arthritis in ankylosing spondylitis.
Fifty one patients with ankylosing spondylitis (AS) were typed for HLA-A, B, C, DR, and DQ antigens. The antigen frequencies were compared with those of a normal population and with a B27 positive control group. All but one of the patients with AS were HLA-B27 positive. A positive linkage disequilibrium between Cw1, Cw2, DR1, and the B27 antigen was observed. Patients with AS showed a significant increase in DQw2 antigen compared with the B27 positive control group. No differences in antigenic frequencies were observed in patients having peripheral arthritis and patients with only axial involvement. Seven out of nine patients (78%) with an erosive peripheral arthritis were DR7 positive, suggesting that DR7 or genes closely linked could be related with a more aggressive peripheral joint involvement in patients with AS.


Yeast Sm-like proteins function in mRNA decapping and decay.
One of the main mechanisms of messenger RNA degradation in eukaryotes occurs by deadenylation-dependent decapping which leads to 5'-to-3' decay. A family of Sm-like (Lsm) proteins has been identified, members of which contain the 'Sm' sequence motif, form a complex with U6 small nuclear RNA and are required for pre-mRNA splicing. Here we show that mutations in seven yeast Lsm proteins (Lsm1-Lsm7) also lead to inhibition of mRNA decapping. In addition, the Lsm1-Lsm7 proteins co-immunoprecipitate with the mRNA decapping enzyme (Dcp1), a decapping activator (Pat1/Mrt1) and with mRNA. This indicates that the Lsm proteins may promote decapping by interactions with the mRNA and the decapping machinery. In addition, the Lsm complex that functions in mRNA decay appears to be distinct from the U6-associated Lsm complex, indicating that Lsm proteins form specific complexes that affect different aspects of mRNA metabolism.