Anthocyanins extracted from black soybean seed coat protect primary cortical neurons against in vitro ischemia.
The present study investigated the neuroprotective effects of anthocyanins extracted from black soybean (cv. Cheongja 3, Glycine max (L.) MERR.) seed coat against oxygen-glucose deprivation (OGD) and glutamate-induced cell death in rat primary cortical neurons. Lactate dehydrogenase (LDH) release and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assays were employed to assess cell membrane damage and viability of primary neurons, respectively. OGD-induced cell death in 7 d in vitro primary cortical neurons was found to be OGD duration-dependent, and approximately 3.5 h of OGD resulted in ≈60% cell death. Treatment with black soybean anthocyanins dose-dependently prevented membrane damage and increased the viability of primary neurons that were exposed to OGD. Glutamate-induced neuronal cell death was dependent on the glutamate concentration at relatively low concentrations and the number of days the cells remained in culture. Interestingly, black soybean anthocyanins did not protect against glutamate-induced neuronal cell death. They did, however, inhibit the excessive generation of reactive oxygen species (ROS) and preserve mitochondrial membrane potential (MMP) in primary neurons exposed to OGD. In agreement with the neuroprotective effect of crude black soybean anthocyanins, purified cyanidin-3-glucoside (C3G), the major component of anthocyanins, also offered dose-dependent neuroprotection against OGD-induced neuronal cell death. Moreover, black soybean C3G markedly prevented excessive generation of ROS and preserved MMP in primary neurons that were exposed to OGD. Collectively, these results suggest that the neuroprotection of primary rat cortical neurons by anthocyanins that were extracted from black soybean seed coat might be mediated through oxidative stress inhibition and MMP preservation but not through glutamate-induced excitotoxicity attenuation.


2.5-year-olds succeed at a verbal anticipatory-looking false-belief task.
Recent research suggests that infants and toddlers succeed at a wide range of non-elicited-response false-belief tasks (i.e., tasks that do not require children to answer a direct question about a mistaken agent's likely behaviour). However, one exception to this generalization comes from verbal anticipatory-looking tasks, which have produced inconsistent findings with toddlers. One possible explanation for these findings is that toddlers succeed when they correctly interpret the prompt as a self-addressed utterance (making the task a non-elicited-response task), but fail when they mistakenly interpret the prompt as a direct question (making the task an elicited-response task). Here, 2.5-year-old toddlers were tested in a verbal anticipatory-looking task that was designed to help them interpret the anticipatory prompt as a self-addressed utterance: the experimenter looked at the ceiling, chin in hand, during and after the prompt. Children gave evidence of false-belief understanding in this task, but failed when the experimenter looked at the child during and after the prompt. These results reinforce claims of robust continuity in early false-belief reasoning and provide additional support for the distinction between non-elicited- and elicited-response false-belief tasks. Three accounts of the discrepant results obtained with these tasks - and of early false-belief understanding more generally - are discussed.


Microbiota modulate sympathetic neurons via a gut-brain circuit.
Connections between the gut and brain monitor the intestinal tissue and its microbial and dietary content1, regulating both physiological intestinal functions such as nutrient absorption and motility2,3, and brain-wired feeding behaviour2. It is therefore plausible that circuits exist to detect gut microorganisms and relay this information to areas of the central nervous system that, in turn, regulate gut physiology4. Here we characterize the influence of the microbiota on enteric-associated neurons by combining gnotobiotic mouse models with transcriptomics, circuit-tracing methods and functional manipulations. We find that the gut microbiome modulates gut-extrinsic sympathetic neurons: microbiota depletion leads to increased expression of the neuronal transcription factor cFos, and colonization of germ-free mice with bacteria that produce short-chain fatty acids suppresses cFos expression in the gut sympathetic ganglia. Chemogenetic manipulations, translational profiling and anterograde tracing identify a subset of distal intestine-projecting vagal neurons that are positioned to have an afferent role in microbiota-mediated modulation of gut sympathetic neurons. Retrograde polysynaptic neuronal tracing from the intestinal wall identifies brainstem sensory nuclei that are activated during microbial depletion, as well as efferent sympathetic premotor glutamatergic neurons that regulate gastrointestinal transit. These results reveal microbiota-dependent control of gut-extrinsic sympathetic activation through a gut-brain circuit.


Typhoid perforation: Post-operative Intensive Care Unit care and outcome.
Typhoid perforation ileitis is a serious complication of typhoid fever, a common and unfortunate health problem in a resource-poor country like Nigeria. Following bowel perforation, treatment is usually by simple closure or bowel resection and anastomosis after adequate aggressive fluid resuscitation and electrolyte correction. Postoperatively, some of these patients do require management in Intensive Care Unit (ICU) on account of sepsis or septic shock and to improve survival. This is a prospective observational study in which 67 consecutive patients who had exploratory laparotomy for typhoid perforation between August 2009 and October 2012 in the main operating theatre of the University College Hospital, Ibadan, were studied. The attending anaesthetists had the freedom of choosing the appropriate anaesthetic drugs depending on the patients' clinical condition. The reason for admission into the ICU, the types of organ support required and outcomes were recorded. Twenty-five patients (37.3%) out of 67 required critical care. Reasons for admission among others included poor respiratory effort, hypotension, septic shock and delayed recovery from anaesthesia. Twenty-one patients (84%) required mechanical ventilation with a mean duration of 2.14 days (range 1-5 days). Fourteen patients required ionotropic support and the length of ICU stay ranged from 1 to 15 days (mean 4.32 days). Nineteen patients (76%) were successfully managed and discharged to the ward while 24% (6 patients) mortality rate was recorded. This study showed high rate of post-operative ICU admission in patients with typhoid perforation with a high demand for critical care involving mechanical ventilation and ionotropic support. In centres that manage patients presenting with typhoid ileitis and perforation, post-operative critical care should be available.


Redox-regulating sirtuins in aging, caloric restriction, and exercise.
The consequence of decreased nicotinamide adenine dinucleotide (NAD(+)) levels as a result of oxidative challenge is altered activity of sirtuins, which, in turn, brings about a wide range of modifications in mammalian cellular metabolism. Sirtuins, especially SIRT1, deacetylate important transcription factors such as p53, forkhead homeobox type O proteins, nuclear factor κB, or peroxisome proliferator-activated receptor γ coactivator 1α (which controls the transcription of pro- and antioxidant enzymes, by which the cellular redox state is affected). The role of SIRT1 in DNA repair is enigmatic, because it activates Ku70 to cope with double-strand breaks, but deacetylation of apurinic/apyrimidinic endonuclease 1 and probably of 8-oxoguanine-DNA glycosylase 1 decreases the activity of these DNA repair enzymes. The protein-stabilizing effects of the NAD+-dependent lysine deacetylases are readily related to housekeeping and redox regulation. The role of sirtuins in caloric restriction (CR)-related longevity in yeast is currently under debate. However, in mammals, it seems certain that sirtuins are involved in many cellular processes that mediate longevity and disease prevention via the effects of CR through the vascular, neuronal, and muscular systems. Regular physical exercise-mediated health promotion also involves sirtuin-regulated pathways including the antioxidant-, macromolecular damage repair-, energy-, mitochondrial function-, and neuronal plasticity-associated pathways. This review critically evaluates these findings and points out the age-associated role of sirtuins.


PCSK9 Inhibitor Use in the Real World: Data From the National Patient-Centered Research Network.
Background PCSK9 (proprotein convertase subtilisin/kexin type 9) inhibitors effectively lower LDL (low-density lipoprotein) cholesterol and have been shown to reduce cardiovascular outcomes in high-risk patients. We used real-world electronic health record data to characterize use of PCSK9 inhibitors, in addition to standard therapies, according to cardiovascular risk status. Methods and Results Data were obtained from 18 health systems with data marts within the National Patient-Centered Clinical Research Network (PCORnet) using a common data model. Participating sites identified >17.5 million adults, of whom 3.6 million met study criteria. Patients were categorized into 3 groups: (1) dyslipidemia, (2) untreated LDL ≥130 mg/dL, and (3) coronary artery disease or coronary heart disease. Demographics, comorbidities, estimated 10-year atherosclerotic cardiovascular disease risk, and lipid-lowering pharmacotherapies were summarized for each group. Participants' average age was 62 years, 50% were female, and 11% were black. LDL cholesterol ranged from 85 to 151 mg/dL. Among patients in groups 1 and 3, 54% received standard lipid-lowering therapies and a PCSK9 inhibitor was prescribed in <1%. PCSK9 inhibitor prescribing was greatest for patients with coronary artery disease or coronary heart disease and, although prescribing increased during the study period, overall PCSK9 inhibitor prescribing was low. Conclusions We successfully used electronic health record data from 18 PCORnet data marts to identify >3.6 million patients meeting criteria for 3 patient groups. Approximately half of patients had been prescribed lipid-lowering medication, but <1% were prescribed PCSK9 inhibitors. PCSK9 inhibitor prescribing increased over time for patients with coronary artery disease or coronary heart disease but not for those with dyslipidemia.


Disclination mediated dynamic recrystallization in metals at low temperature.
Recrystallization is one of the most important physical phenomena in condensed matter that has been utilized for materials processing for thousands of years in human history. It is generally believed that recrystallization is thermally activated and a minimum temperature must be achieved for the necessary atomic mechanisms to occur. Here, using atomistic simulations, we report a new mechanism of dynamic recrystallization that can operate at temperature as low as T = 10 K in metals during deformation. In contrast to previously proposed dislocation-based models, this mechanism relies on the generation of disclination quadrupoles, which are special defects that form during deformation when the grain boundary migration is restricted by structural defects such as triple junctions, cracks or obstacles. This mechanism offers an alternative explanation for the grain refinement in metals during severe plastic deformation at cryogenic temperature and may suggest a new method to tailor the microstructure in general crystalline materials.


Barriers to adoption of recent technology in cervical screening.
The Pap smear is one of the modern success stories in the field of preventive medicine. Since its introduction as a screening test, there has been a dramatic reduction in the incidence of cervical cancer. However, the search for a better screening test continues. The new technologies, including liquid-based cytology (LBC), Human Papilloma Virus (HPV) testing and automated or machine-assisted screening have been introduced. However, there is continuous debate about whether society's limited resources are better spent on reaching the underserved rather than on these technologies. Another question is whether these technologies create yet another kind of disparity in delivering preventive care. For example, despite the wide use of LBC (99% of tests submitted to our laboratory are LBC), conventional Pap smears are still used to screen/follow up some women. It is not clear why some providers continue to prefer conventional smear over LBC and what are the barriers for adopting LBC in cervical cancer screening. We hypothesize the lower cost of conventional compared to LBC Pap testing, patient's lower socio-economic indices, a patient's medical history and provider's subspecialty/training all appear to play a role in the choice of using conventional Pap testing rather than LBC. Unintentionally, this choice results in repeat testing, delayed treatment and potentially higher costs than intended. The ultimate goal of this review article is to understand and explore possible barriers and disparities to adopting new technology in cancer screening.


Natural history and outcomes in drug-induced autoimmune hepatitis.
Drug-induced autoimmune hepatitis (DIAIH) remains poorly characterized. Our aim was to assess natural history and outcomes in DIAIH. This was a retrospective cohort study. Eighty-two patients with autoimmune hepatitis (AIH) were identified, 11 (13.4%) with DIAIH, implicated drugs being nitrofurantoin (n = 4), statins (n = 4), herbal remedies (n = 2) and diclofenac (n = 1). Female sex, acute onset, elevated serum globulins/immunoglobulin G, fibrosis stage (Ishak), cirrhosis at onset, moderate-severe portal inflammation, interface and lobular hepatitis, remission, relapse and poor outcome were similar in those with DIAIH and AIH (P > 0.05). The former were however more likely to be aged 60 years or more and take longer to relapse on immunosuppression discontinuation (P = <0.05). On Kaplan-Meier analysis, probability of poor outcome was similar in those with DIAIH and AIH (log-rank test, 0.339). On comparing those with (n = 4) and without nitrofurantoin (n = 7) DIAIH, the former were older, had longer duration of drug use prior to DIAIH diagnosis, higher fibrosis stage and were less likely to relapse upon immunosuppression discontinuation. Approximately 15% of patients with AIH have DIAIH with similar outcomes, although the latter are older with a propensity for late relapse, mandating long-term follow up.


A protein assembly mediates Xist localization and gene silencing.
Nuclear compartments have diverse roles in regulating gene expression, yet the molecular forces and components that drive compartment formation remain largely unclear1. The long non-coding RNA Xist establishes an intra-chromosomal compartment by localizing at a high concentration in a territory spatially close to its transcription locus2 and binding diverse proteins3-5 to achieve X-chromosome inactivation (XCI)6,7. The XCI process therefore serves as a paradigm for understanding how RNA-mediated recruitment of various proteins induces a functional compartment. The properties of the inactive X (Xi)-compartment are known to change over time, because after initial Xist spreading and transcriptional shutoff a state is reached in which gene silencing remains stable even if Xist is turned off8. Here we show that the Xist RNA-binding proteins PTBP19, MATR310, TDP-4311 and CELF112 assemble on the multivalent E-repeat element of Xist7 and, via self-aggregation and heterotypic protein-protein interactions, form a condensate1 in the Xi. This condensate is required for gene silencing and for the anchoring of Xist to the Xi territory, and can be sustained in the absence of Xist. Notably, these E-repeat-binding proteins become essential coincident with transition to the Xist-independent XCI phase8, indicating that the condensate seeded by the E-repeat underlies the developmental switch from Xist-dependence to Xist-independence. Taken together, our data show that Xist forms the Xi compartment by seeding a heteromeric condensate that consists of ubiquitous RNA-binding proteins, revealing an unanticipated mechanism for heritable gene silencing.


Fatigue in patients with juvenile idiopathic arthritis: A systematic review of the literature.
To perform a systematic review of the current literature on studies related to fatigue in children with JIA. We studied the measurements that were used to assess fatigue and we focused on three outcome measurements, namely, (1) the prevalence of fatigue in JIA patients, (2) the determinants of and associations with fatigue in JIA patients, and (3) the impact of fatigue in JIA patients on daily life. A search was conducted in the electronic databases Pubmed and Embase from January 1, 2000 until August 27, 2015. The quality in prognostic factors (QUIPS) tool was used to assess the risk of bias (ROB) in the selected studies, focused on the outcome fatigue. Of all, two authors independently judged the ROB. A total of 15 studies were included in this review. To assess fatigue, two unidimensional and three multidimensional scales were used, which hampered comparison. Fatigue was reported to be present in 60-76% of the patients with JIA and was related to time of day, disease activity, pain, psychosocial factors, and sleep. Minor consequences for daily life were found, though this was not studied extensively. Fatigue is common in patients with JIA. The cause reflects a complex interplay of different factors. Based on results from this review and its body of knowledge, a conceptual model for fatigue in patients with JIA is proposed. Consensus is needed for future studies on how to assess fatigue. We promote the use of a combination of unidimensional and validated multidimensional measurements.


Anti-cyclic citrullinated peptide antibodies and rheumatoid factor in Sjögren's syndrome.
The purpose of this study was to evaluate the prevalence and clinical significance of anti-cyclic citrullinated peptide antibodies (anti-CCP-Abs), IgM and IgA rheumatoid factors (RFs) in primary Sjögren's Syndrome (pSS). We compared clinical and serological characteristics of 31 pSS and 31 Rheumatoid Arthritis (RA) patients. Both, anti-CCP-Abs and RFs (IgM, IgA) directed against Fc determinants of IgG from humans and rabbit were detected by enzyme-linked immunosorbent assay (ELISA). We included 31 blood donors as control group for the evaluation of RFs and anti-CCP-Abs. Nine (29%) pSS patients presented arthritis, and 10 (32,3%) RA patients also had secondary Sjögren's syndrome (sSS) RESULTS: IgM and IgA RFs prevalence was similar in pSS and RA, whichever the antigene (Human or Rabbit IgG) used. However, RA patients with sSS showed a tendency to present more often RF positivity, longer disease duration and higher ESR and CRP when compared with pSS patients with arthritis. Anti-CCP-Abs were detected in 64,5% of RA patients and in only 6,9% of pSS patients (p<0,0005). Anti-CCP-Abs were more often positive in RA patients with sSS (RA/sSS) (8 patients, 80%) than in RA patients without sSS (18 patients, 58,1%), and were absent in pSS patients with arthritis. RF-positive pSS patients presented more often pulmonary involvement and higher inflammatory parameters, and less often neuropathy compared to RF-negative patients. In controls, anti-CCP-Abs were absent and RFs were negligible. Anti-CCP-Abs were detected in only a few pSS patients, none of whom presented arthritis, which contrasts with the high frequency of these antibodies in RA/sSS. These results suggest that anti-CCP-Abs could be useful in the distinction between pSS and RA with sSS. Although not useful for the differential diagnosis between RA and pSS, RFs may have a prognostic role in pSS.


Identification of genes controlling germ cell migration and embryonic gonad formation in Drosophila.
Gonadogenesis in the Drosophila embryo is a complex process involving numerous cellular migratory steps and cell-cell interactions. The mechanisms guiding germ cells to move through, recognize and adhere to specific cell types are poorly understood. In order to identify genes that are required for these processes, we have conducted an extensive mutagenesis of the third chromosome and screened for mutations disrupting germ cell migration at any point in embryonic development. Phenotypic analysis of these mutants demonstrates that germ cell migration can be broken down into discrete developmental steps, with each step requiring a specific set of genes. Many of these genes are involved in the development of gonadal mesoderm, the tissue that associates with germ cells to form the embryonic gonad. Moreover, mutations that we isolated affecting embryonic patterning as well as germ cell migration suggest that the origin of gonadal mesoderm lies within the eve domain of the developing mesoderm.


Identification of Outer Membrane Proteins Altered in Response to UVC-Radiation in Vibrio parahaemolyticus and Vibrio alginolyticus.
Vibrio parahaemolyticus and V. alginolyticus, marine foodborne pathogens, were treated with UVC-radiation (240 J/m(2)) to evaluate alterations in their outer membrane protein profiles. Outer membrane protein patterns of UVC-irradiated bacteria were found altered when analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Altered proteins were identified by mass spectrometry (MS and MS/MS) and analysis revealed that OmpW, OmpA, Long-chain fatty acid transport protein, Outer membrane receptor protein, Putative uncharacterized protein VP0167, Maltoporin (lamB), Polar flagellin B/D, Agglutination protein Peptidoglycan-associated lipoprotein and MltA-interacting protein MipA were appeared, thereby they can be considered as UVC-stress proteins in some vibrios. In addition, expression of OmpK decreased to non-detectable level. Furthermore, we observed a decrease or an increase in the expression level of other outer membrane proteins.


A pragmatic randomized controlled trial of a guided self-help intervention versus a waiting list control in a routine primary care mental health service.
OBJECTIVES. To evaluate the effectiveness of a two session guided self-help (GSH) intervention provided by primary care graduate mental health workers (PCGMHWs) in a primary care mental health service. DESIGN. Pragmatic randomized trial, with a wait list control design. METHOD. Patients presenting with significant anxiety and depression problems were given one or more self-help booklets at screening and randomly allocated to an immediate (ITG) or delayed treatment group (DTG). Following this, a two-session GSH intervention was provided by one of two PCGMHWs, with a review session to decide on the need for further intervention. The DTG began the intervention 8 weeks after the screening and the primary outcome was Clinical Outcomes in Routine Evaluation - Outcome Measure (CORE-OM) scores after 8 weeks. RESULTS. A total of 63 patients were allocated to the ITG, 59 to the DTG. Analysis of covariance, carried out on an intention to treat basis, showed a significant treatment effect, F(1,98) = 15, p < .001, and a comparison of means at 8 weeks showed a significant difference, t(116) = 2.1 (95% CI [1.1, 5.9]), p= .042 with an effect size, d= 0.375. Taking the two groups together, CORE-OM scores for patients who completed the intervention reduced between screening and the review session by an average of 7.9 (95% CI [6.3, 9.5]), effect size of 1.2. Between screening and the review session, 47% showed a reliable and clinically significant improvement. CONCLUSIONS. The study provides some support for the effectiveness of a two-session GSH intervention and a stepped-care service model.


Improving methicillin-resistant Staphylococcus aureus surveillance and reporting in intensive care units.
Routine culturing of patients in intensive care units (ICUs) for methicillin-resistant Staphylococcus aureus (MRSA) identifies unrecognized carriers and facilitates timely isolation. However, the benefit of surveillance in detecting prevalent and incident carriers likely varies among ICUs. In addition, many assessments underestimate the incidence of acquisition by including prevalent carriers in the at-risk population. We performed a retrospective cohort study using accurate at-risk populations to evaluate the range of benefit of admission and weekly surveillance cultures in detecting otherwise unrecognized MRSA in 12 ICUs in 5 states. We assessed 142 ICU-months. Among the 12 ICUs, the admission prevalence of imported MRSA was 5%-21%, with admission surveillance providing 30%-135% increases in rates of detection. The monthly hospital-associated incidence was 2%-6%, with weekly surveillance providing 7%-157% increases in detection. The common practice of reporting incidence using the total number of patients or total patient-days underestimated incidence by one-third. Surgical ICUs had lower MRSA importation but higher MRSA incidence. Overall, routine surveillance prevented the misclassification of 17% (unit range, 11%-29%) of "incident" carriers, compared with clinical cultures, and increased precaution days by 18% (unit range, 11%-91%). Routine surveillance significantly increases the detection of MRSA, but this benefit is not uniform across ICUs, even with high compliance and the use of correct denominators.


Turnover in close friendships.
Humans are social animals and the interpersonal bonds formed between them are crucial for their development and well being in a society. These relationships are usually structured into several layers (Dunbar's layers of friendship) depending on their significance in an individual's life with closest friends and family being the most important ones taking major part of their time and communication effort. However, we have little idea how the initiation and termination of these relationships occurs across the lifespan. Mobile phones, in particular, have been used extensively to shed light on the different types of social interactions between individuals and to explore this, we analyse a national cellphone database to determine how and when changes in close relationships occur in the two genders. In general, membership of this inner circle of intimate relationships is extremely stable, at least over a three-year period. However, around 1-4% of alters change every year, with the rate of change being higher among 17-21 year olds than older adults. Young adult females terminate more of their opposite-gender relationships, while older males are more persistent in trying to maintain relationships in decline. These results emphasise the variability in relationship dynamics across age and gender, and remind us that individual differences play an important role in the structure of social networks. Overall, our study provides a holistic understanding of the dynamic nature of close relationships during different stages of human life.


Role of fine needle aspiration and frozen section in determining the extent of thyroidectomy.
The routine use of fine needle aspiration (FNA) and frozen section (FS) in the management of a thyroid nodule is controversial and needs to be evaluated on an institution to institution basis. Our aim was to determine the role of FNA and FS in determining the extent of thyroidectomy. We performed a comparative study of FNA and FS examination of all patients presenting with nodular thyroid disease between September 2002 and December 2005. Data were collected on a proforma by reviewing FNA, FS and histopathological reports. Data were analyzed on SPSS 11. Sensitivity, specificity, accuracy, positive predictive value and negative predictive values were calculated. We included 44 patients with preoperative FNA, intraoperative FS examination and final histopathology reports available. We excluded patients with local invasion and distant metastases. Final histopathological report was taken as gold standard. FNA reported 8 benign, 7 papillary carcinoma, 22 follicular neoplasm, 1 medullary and 6 suspicious lesions. On final pathology there were 16 benign and 28 malignant cases. Thus a total of 20 carcinomas were missed by FNA. When routine FS was done, a total of ten patients who had malignancy were missed. Both FNA and FS have high specificity for diagnosis of thyroid cancer but lacked sensitivity at our institution. This is mainly because of high false negative results.


Functions of Stress-Induced Lipid Droplets in the Nervous System.
Lipid droplets are highly dynamic intracellular organelles that store neutral lipids such as cholesteryl esters and triacylglycerols. They have recently emerged as key stress response components in many different cell types. Lipid droplets in the nervous system are mostly observed in vivo in glia, ependymal cells and microglia. They tend to become more numerous in these cell types and can also form in neurons as a consequence of ageing or stresses involving redox imbalance and lipotoxicity. Abundant lipid droplets are also a characteristic feature of several neurodegenerative diseases. In this minireview, we take a cell-type perspective on recent advances in our understanding of lipid droplet metabolism in glia, neurons and neural stem cells during health and disease. We highlight that a given lipid droplet subfunction, such as triacylglycerol lipolysis, can be physiologically beneficial or harmful to the functions of the nervous system depending upon cellular context. The mechanistic understanding of context-dependent lipid droplet functions in the nervous system is progressing apace, aided by new technologies for probing the lipid droplet proteome and lipidome with single-cell type precision.


The chevron osteotomy for correction of hallux valgus. Comparison of findings after two and five years of follow-up.
The chevron osteotomy, an accepted method for the correction of mild and moderate hallux valgus, is generally advocated for patients younger than the age of fifty years. The purposes of this prospective study were to compare the short-term (two-year) and intermediate-term (five-year) results of this operation with respect to patient satisfaction, flexion and extension of the metatarsophalangeal joint, maintenance of correction, and development of arthrosis and to determine whether the effectiveness of the procedure was limited by age. Between April 1991 and September 1992, the chevron osteotomy was performed for the treatment of mild-to-moderate hallux valgus deformity in sixty-six consecutive feet. Forty-three patients (fifty-seven feet) were available for follow-up at both two and five years postoperatively. The two-year and five-year clinical assessments were based on the American Orthopaedic Foot and Ankle Society's hallux-metatarsophalangeal-interphalangeal scale. Between the two-year and five-year follow-up evaluations, there was only a minimal change in overall patient satisfaction, and the average score on the hallux-metatarsophalangeal-interphalangeal scale was unchanged. The passive range of motion of the first metatarsophalangeal joint decreased between the preoperative assessment and the two-year follow-up evaluation and was unchanged at the five-year follow-up evaluation. Radiographic evaluation showed no changes in the hallux valgus or intermetatarsal angle between the two-year and five-year evaluations, although the number of feet with arthrosis of the metatarsophalangeal joint increased slightly, from eight to eleven. Patients aged fifty years or older did as well as younger patients. At these two follow-up periods, the chevron osteotomy was found to be a reliable procedure for the correction of mild and moderate hallux valgus deformity, and outcome did not differ on the basis of age.


Increasing cropping system diversity balances productivity, profitability and environmental health.
Balancing productivity, profitability, and environmental health is a key challenge for agricultural sustainability. Most crop production systems in the United States are characterized by low species and management diversity, high use of fossil energy and agrichemicals, and large negative impacts on the environment. We hypothesized that cropping system diversification would promote ecosystem services that would supplement, and eventually displace, synthetic external inputs used to maintain crop productivity. To test this, we conducted a field study from 2003-2011 in Iowa that included three contrasting systems varying in length of crop sequence and inputs. We compared a conventionally managed 2-yr rotation (maize-soybean) that received fertilizers and herbicides at rates comparable to those used on nearby farms with two more diverse cropping systems: a 3-yr rotation (maize-soybean-small grain + red clover) and a 4-yr rotation (maize-soybean-small grain + alfalfa-alfalfa) managed with lower synthetic N fertilizer and herbicide inputs and periodic applications of cattle manure. Grain yields, mass of harvested products, and profit in the more diverse systems were similar to, or greater than, those in the conventional system, despite reductions of agrichemical inputs. Weeds were suppressed effectively in all systems, but freshwater toxicity of the more diverse systems was two orders of magnitude lower than in the conventional system. Results of our study indicate that more diverse cropping systems can use small amounts of synthetic agrichemical inputs as powerful tools with which to tune, rather than drive, agroecosystem performance, while meeting or exceeding the performance of less diverse systems.


Establishment and characterization of conditionally immortalized cells from the mouse urogenital ridge.
Cell cultures from the urogenital ridge have been established to facilitate the study of the regulation and downstream interactions of Sry in mammalian sex determination. Cells have been explanted from transgenic mice carrying a temperature sensitive SV40 large T-antigen, and established in ongoing cultures. Analysis of the cells in these cultures at the electron microscope level reveals multiple cell types that compare to the cell types found in vivo during this period of development. Primordial germ cells, that are simultaneously explanted in the course of these experiments, also survive in culture. The explants undergo a morphogenetic organization into branching cord-like structures when cells are trypsinized and plated in extracellular matrix (Matrigel). We analyzed the expression of a number of molecular markers of the fetal gonad during monolayer culture, during in vitro morphogenesis in Matrigel, and in clonal lines derived from the complex explants. This analysis included Sry which is found to be expressed in some cultures from XY urogenital ridges that have been maintained for as long as 8 months.


Hrd1 suppresses Nrf2-mediated cellular protection during liver cirrhosis.
Increased endoplasmic reticulum (ER) stress and reactive oxygen species (ROS) are the salient features of end-stage liver diseases. Using liver tissues from liver cirrhosis patients, we observed up-regulation of the XBP1-Hrd1 arm of the ER stress response pathway and down-regulation of the Nrf2-mediated antioxidant response pathway. We further confirmed this negative regulation of Nrf2 by Hrd1 using Hrd1 conditional knockout mice. Down-regulation of Nrf2 was a surprising result, since the high levels of ROS should have inactivated Keap1, the primary ubiquitin ligase regulating Nrf2 levels. Here, we identified Hrd1 as a novel E3 ubiquitin ligase responsible for compromised Nrf2 response during liver cirrhosis. In cirrhotic livers, activation of the XBP1-Hrd1 arm of ER stress transcriptionally up-regulated Hrd1, resulting in enhanced Nrf2 ubiquitylation and degradation and attenuation of the Nrf2 signaling pathway. Our study reveals not only the convergence of ER and oxidative stress response pathways but also the pathological importance of this cross-talk in liver cirrhosis. Finally, we showed the therapeutic importance of targeting Hrd1, rather than Keap1, to prevent Nrf2 loss and suppress liver cirrhosis.


Managing Spaghetti Syndrome in Critical Care With a Novel Device: A Nursing Perspective.
Managing "spaghetti syndrome," the tangle of therapeutic cables, tubes, and cords at patients' bedsides, can be challenging. To assess nurses' perceptions of the effectiveness of a novel banding device in management of spaghetti syndrome. A simple color-coded elastomeric banding strap with ribbed flaps was attached to bed rails of adult critical care patients to help organize therapeutic cables, tubes, wires, and cords. Nurses were surveyed before and after use of the bands and after the nursing shift to assess the burden of spaghetti syndrome and the effectiveness of using the bands. Use of the bands decreased the time spent untangling cords, reduced the frequency of contact of tubing with the floor, and diminished disruptions in care. Use of a simple flexible latex-free elastomeric band may help organize therapeutic tubing at patients' bedsides and may promote improvements in nursing care.


Incidence and pattern of direct blunt neurovascular injury associated with trauma to the skull base.
Skull base fractures are often associated with potentially devastating injuries to major neural arteries in the head and neck, but the incidence and pattern of this association are unknown. Between April and September 2002, 1738 Level 1 trauma patients were admitted to St. Joseph's Hospital and Medical Center in Phoenix, Arizona. Among them, a skull base fracture was diagnosed in 78 patients following computed tomography (CT) scans. Seven patients had no neurovascular imaging performed and were excluded. Altogether, 71 patients who received a diagnosis of skull base fractures after CT and who also underwent a neurovascular imaging study were included (54 men and 17 women, mean age 29 years, range 1-83 years). Patients underwent CT angiography, magnetic resonance angiography, or digital subtraction angiography of the head and craniovertebral junction, or combinations thereof. Nine neurovascular injuries were identified in six (8.5%) of the 71 patients. Fractures of the clivus were very likely to be associated with neurovascular injury (p < 0.001). A high risk of neurovascular injury showed a strong tendency to be associated with fractures of the sella turcica-sphenoid sinus complex (p = 0.07). The risk of associated blunt neurovascular injury appears to be significant in Level 1 trauma patients in whom a diagnosis of skull base fracture has been made using CT. The incidence of neurovascular trauma is particularly high in patients with clival fractures. The authors recommend neurovascular imaging for Level 1 trauma patients with a high-risk fracture pattern of the central skull base to rule out cerebrovascular injuries.


Population impact of direct-acting antiviral treatment on new presentations of hepatitis C-related decompensated cirrhosis: a national record-linkage study.
Population-based studies demonstrating the clinical impact of interferon-free direct-acting antiviral (DAA) therapies are lacking. We examined the impact of the introduction of DAAs on HCV-related decompensated cirrhosis (DC) through analysis of population-based data from Scotland. Through analysis of national surveillance data (involving linkage of HCV diagnosis and clinical databases to hospital and deaths registers), we determined i) the scale-up in the number of patients treated and achieving a sustained viral response (SVR), and ii) the change in the trend of new presentations with HCV-related DC, with the introduction of DAAs. Approximately 11 000 patients had been treated in Scotland over the 8-year period 2010/11 to 2017/18. The scale-up in the number of patients achieving SVR between the pre-DAA and DAA eras was 2.3-fold overall and 5.9-fold among those with compensated cirrhosis (the group at immediate risk of developing DC). In the pre-DAA era, the annual number of HCV-related DC presentations increased 4.6-fold between 2000 (30) and 2014 (142). In the DAA era, presentations decreased by 51% to 69 in 2018 (and by 67% among those with chronic infection at presentation), representing a significant change in trend (rate ratio 0.88, 95% CI 0.85 to 0.90). With the introduction of DAAs, an estimated 330 DC cases had been averted during 2015-18. National scale-up in interferon-free DAA treatment is associated with the rapid downturn in presentations of HCV-related DC at the population-level. Major progress in averting HCV-related DC in the short-term is feasible, and thus other countries should strive to achieve the same.


Is the prevalence of overactive bladder overestimated? A population-based study in Finland.
In earlier studies, one in six adults had overactive bladder which may impair quality of life. However, earlier studies have either not been population-based or have suffered from methodological limitations. Our aim was to assess the prevalence of overactive bladder symptoms, based on a representative study population and using consistent definitions and exclusions. The aim of the study was to assess the age-standardized prevalence of overactive bladder defined as urinary urgency, with or without urgency incontinence, usually with urinary frequency and nocturia in the absence of urinary tract infection or other obvious pathology. In 2003-2004, a questionnaire was mailed to 6,000 randomly selected Finns aged 18-79 years who were identified from the Finnish Population Register Centre. Information on voiding symptoms was collected using the validated Danish Prostatic Symptom Score, with additional frequency and nocturia questions. Corrected prevalence was calculated with adjustment for selection bias due to non-response. The questionnaire also elicited co-morbidity and socio-demographic information. Of the 6,000 subjects, 62.4% participated. The prevalence of overactive bladder was 6.5% (95% CI, 5.5% to 7.6%) for men and 9.3% (CI, 7.9% to 10.6%) for women. Exclusion of men with benign prostatic hyperplasia reduced prevalence among men by approximately one percentage point (to 5.6% [CI, 4.5% to 6.6%]). Among subjects with overactive bladder, urgency incontinence, frequency, and nocturia were reported by 11%, 23%, and 56% of men and 27%, 38%, and 40% of women, respectively. However, only 31% of men and 35% of women with frequency, and 31% of subjects of both sexes with nocturia reported overactive bladder. Our results indicate a prevalence of overactive bladder as low as 8% suggesting that, in previous studies, occurrence has been overestimated due to vague criteria and selected study populations regarding age distribution and low participation.


The effect of noncognitive traits on health behaviours in adolescence.
This paper investigates the relationship between personality traits and health behaviours in adolescence using a large and recent cohort study. In particular, we investigate the impact of locus of control, self-esteem and work ethics at ages 15-16 years on the incidence of health behaviours such as alcohol consumption, cannabis and other drug use, unprotected and early sexual activity and sports and physical activity. We use matching methods to control for a very rich set of adolescent and family characteristics, and we find that personality traits do affect health behaviours. In particular, individuals with external locus of control, low self-esteem or with low levels of work ethics seem more likely in engage in risky health behaviours.


Community-Supported Agriculture as a Dietary and Health Improvement Strategy: A Narrative Review.
This narrative review summarizes the literature regarding community-supported agriculture (CSA) with a focus on its use as a dietary and health improvement strategy. CSA members are typically women, white, highly educated, and affluent. The majority of members are motivated to participate in CSA by a concern for the environment and a desire for locally grown, high-quality, and organic produce. Numerous studies have provided evidence of the economic, community, environmental, and food quality related benefits of CSAs. A substantial body of literature has also explored the CSA member experience and has found that members are generally very satisfied, but membership turnover rates are often high. Research regarding the association between dietary intake and health is more limited and mostly descriptive in nature. CSA members often report increased consumption and variety of fruits and vegetables, changes in the household food environment, and changes in meal patterns. A small number of anecdotal reports also support the association between CSA participation and improved health status. However, there is a dearth of experimental research in this area, and results of these studies are mixed. Future research opportunities include longitudinal studies to evaluate repeat CSA participation and the long-term sustainability of CSA-related dietary and health changes. In addition, research is needed to address some of the methodologic limitations of the current research with regard to survey tools, generalizability of results, self-reporting bias, and CSA member support.


Two-level burst fractures: clinical evaluation and treatment options.
Two-level burst fractures are rare. In a series of 180 surgically treated spinal fracture-dislocations, seven had such injuries, with an incidence of 3.9%. Four had fracture sites without contiguity: C4-T12 (one), L1-L4 (one), L2-L4 (one), and L2-L5 (one); and three with contiguity: T12-L1 (one), L1-L2 (one), and L2-L3 (one). L2 was the most frequently involved site, accounting for four in seven. Falling from height was the most common mechanism of injury, accounting for four in seven. Five in seven patients (71%) sustained multiple injuries. Chest traumas and extremity fractures were the ones most frequently associated. All of these patients had incomplete neurologic deficits at initial presentation. In the four discontiguous bursts, the neurologic levels corresponded to the cephalic ones. Six patients had follow-up periods of more than 2 years. Transpediculate systems were used in five, and at follow-up, two had screw breakages. In this series, the average neurologic recovery was 1.3 grades on the Frankel scale. In conclusion, it is mandatory to have a thorough organ system review when such patients are first seen. Then each fracture site would be judged seperately as either a stable or unstable burst preoperatively. Every effort should be made to treat any unstable segment via anterior, posterior, or combined approaches.


High frequency of deletion mutations in p53 gene from squamous cell lung cancer patients in Taiwan.
Lung cancer is the leading and second-leading cause of cancer deaths among women and men in Taiwan, respectively. However, the molecular mechanisms involved in lung tumorigenesis in Taiwan remain poorly defined. A study that analyzed the mutation spectrum of the p53 tumor suppressor gene in 35 female lung cancer patients in Hong Kong showed that a high proportion of the mutations observed were deletions, suggesting the possible involvement of a distinct mutagenic factor(s) in Chinese female lung cancer patients (Y. Takagi et al., Cancer Res., 55: 5354-5357, 1995). Therefore, to gain insight into the role of the p53 tumor suppressor gene and possible etiological factors in lung tumorigenesis in Taiwan, we investigated the mutation spectra of exons 4-11 in the p53 tumor suppressor gene of 60 lung cancer patients in Taiwan. These data were also correlated with clinical pathological characteristics of patients. Lung tumors were surgically resected, genomic DNA was isolated, and their mutation spectra were examined using PCR/single-strand conformational polymorphism analysis and direct sequencing. The frequency of p53 gene mutation was 18% (11 of 60). However, distinct patterns of p53 gene mutation were observed. Seven of 11 mutations detected (64%) were deletions of 1-12 bp at G:C bp or at bp in the immediate vicinity of repetitive sequences and/or tandem repeat sequences. In addition, two patients (2 of 11, 18%) exhibited nonsense mutations. In contrast to the frequent occurrence of missense mutations in the p53 gene reported in the literature, the majority (82%) of the mutations in lung cancer patients in Taiwan were nonmissense mutations, ie., deletions and nonsense mutations. Immunohistochemical staining indicated that p53 mutations including non-in-frame deletions and nonsense mutations all resulted in no expression of p53 protein. Notably, mutations occurred more frequently in patients suffering from squamous cell carcinoma (SQ). Nine of 31 SQ patients (29%) exhibited deletions or nonsense mutations, suggesting that deletions and nonsense mutations in the p53 gene are involved in the formation of SQ in Taiwan. In addition, mutations occurred more frequently in patients with stage III or IV lung cancer. However, mutations were not correlated with patients' smoking habits. Our data suggest that p53 gene mutation involved in the formation of SQ and distinct environmental factor(s) and/or genetic factor(s) that induced specific short deletions in repeat sequences may be involved in lung tumorigenesis in Taiwan.


Endodontic Periapical Lesion: An Overview on the Etiology, Diagnosis and Current Treatment Modalities.
Nonsurgical and surgical endodontic treatments have a high success rate in the treatment and prevention of apical periodontitis when carried out according to standard and accepted clinical principles. Nevertheless, endodontic periapical lesions remain in some cases, and further treatment should be considered when apical periodontitis persists. Although several treatment modalities have been proposed for endodontically treated teeth with persistent apical periodontitis, there is a need for less invasive methods with more predictable outcomes. The advantages and shortcomings of existing approaches for the diagnosis and treatment of endodontic periradicular lesions are discussed in this review.


Evaluation of an automated knowledge based treatment planning system for head and neck.
This study evaluated an automated inverse treatment planning algorithm, Pinnacle Auto-Planning (AP), and compared automatically generated plans with historical plans in a large cohort of head and neck cancer patients. Fifty consecutive patients treated with volumetric modulated arc therapy (Eclipse, Varian Medical System, Palo Alto, CA) for head and neck were re-planned with AP version 9.10. Only one single cycle of plan optimization using one single template was allowed for AP. The dose to the planning target volumes (PTV's; 3-4 dose levels), the organs at risk (OAR's) and the effective working time for planning was evaluated. Additionally, two experienced radiation oncologists blind-reviewed and ranked 10 plans. Dose coverage and dose homogeneity of the PTV were significantly improved with AP, however manually optimized plans showed significantly improved dose conformity. The mean dose to the parotid glands, oral mucosa, swallowing muscles, dorsal neck tissue and maximal dose to the spinal cord were significantly reduced with AP. In 64 % of the plans, the mean dose to any OAR (spinal cord excluded) was reduced by >20 % with AP in comparison to the manually optimized plans. In 12 % of the plans, the manually optimized plans showed reduced doses by >20 % in at least one OAR. The experienced radiation oncologists preferred the AP plan and the clinical plan in 80 and 20 % of the cases, respectively. The average effective working time was 3.8 min ± 1.1 min in comparison to 48.5 min ± 6.0 min using AP compared to the manually optimized plans, respectively. The evaluated automated planning algorithm achieved highly consistent and significantly improved treatment plans with potentially clinically relevant OAR sparing by >20 % in 64 % of the cases. The effective working time was substantially reduced with Auto-Planning.


Auditory rehabilitation of patients with neurofibromatosis Type 2 by using cochlear implants.
The aim of this study was to determine whether patients with neurofibromatosis Type 2 (NF2) who have intact ipsilateral cochlear nerves can have open-set speech discrimination following cochlear implantation. Records of 7 patients with documented NF2 were reviewed to determine speech discrimination outcomes following cochlear implantation. Outcomes were measured using consonant-nucleus-consonant words and phonemes; Hearing in Noise Test sentences in quiet; and City University of New York sentences in quiet and in noise. Preoperatively, none of the patients had open-set speech discrimination. Five of the 7 patients had previously undergone excision of ipsilateral vestibular schwannoma (VS). One of the patients who received a cochlear implant had received radiation therapy for ipsilateral VS, and another was undergoing observation for a small ipsilateral VS. Following cochlear implantation, 4 of 7 patients with NF2 had open-set speech discrimination following cochlear implantation during extended follow-up (15-120 months). Two of the 3 patients without open-set speech understanding had a prolonged period between ipsilateral VS resection and cochlear implantation (120 and 132 months), and had cochlear ossification at the time of implantation. The other patient without open-set speech understanding had good contralateral hearing at the time of cochlear implantation. Despite these findings, 6 of the 7 patients were daily users of their cochlear implants, and the seventh is an occasional user, indicating that all of the patients subjectively gained some benefit from their implants. Cochlear implantation can provide long-term auditory rehabilitation, with open-set speech discrimination for patients with NF2 who have intact ipsilateral cochlear nerves. Factors that can affect implant performance include the following: 1) a prolonged time between VS resection and implantation; and 2) cochlear ossification.


Screening vaccine formulations for biological activity using fresh human whole blood.
Understanding the relevant biological activity of any pharmaceutical formulation destined for human use is crucial. For vaccine-based formulations, activity must reflect the expected immune response, while for non-vaccine therapeutic agents, such as monoclonal antibodies, a lack of immune response to the formulation is desired. During early formulation development, various biochemical and biophysical characteristics can be monitored in a high-throughput screening (HTS) format. However, it remains impractical and arguably unethical to screen samples in this way for immunological functionality in animal models. Furthermore, data for immunological functionality lag formulation design by months, making it cumbersome to relate back to formulations in real-time. It is also likely that animal testing may not accurately reflect the response in humans. For a more effective formulation screen, a human whole blood (hWB) approach can be used to assess immunological functionality. The functional activity relates directly to the human immune response to a complete formulation (adjuvant/antigen) and includes adjuvant response, antigen response, adjuvant-modulated antigen response, stability, and potentially safety. The following commentary discusses the hWB approach as a valuable new tool to de-risk manufacture, formulation design, and clinical progression.


Proton and phosphorus magnetic resonance spectroscopy of a mouse model of Alzheimer's disease.
The development of new diagnostic criteria for Alzheimer's disease (AD) requires new in vivo markers reflecting early pathological changes in the brain of patients. Magnetic resonance (MR) spectroscopy has been shown to provide useful information about the biochemical changes occurring in AD brain in vivo. The development of numerous transgenic mouse models of AD has facilitated the evaluation of early biomarkers, allowing researchers to perform longitudinal studies starting before the onset of the pathology. In addition, the recent development of high-field animal scanners enables the measurement of brain metabolites that cannot be reliably quantified at lower magnetic fields. In this report, we studied a new transgenic mouse model of AD, the 5xFAD model, by in vivo proton and phosphorus MR spectroscopy. This model, which is characterized by an early-onset and a robust amyloid pathology, developed changes in the neurochemical profile, which are typical in the human disease, i.e., an increase in myo-inositol and a decrease in N-acetylaspartate concentrations, as early as in the 40th week of age. In addition, a significant decrease in the γ-aminobutyrate concentration was observed in transgenic mice at this age compared to controls. The pseudo-first-order rate constant of the creatine kinase reaction as well as relative concentrations of phosphorus-containing metabolites were not changed significantly in the 36 and 72-week old transgenic mice. Overall, these results suggest that mitochondrial activity in the 5 × FAD mice is not substantially affected but that the model is relevant for studying early biomarkers of AD.


Do we need the third dimension? Quantifying the effect of the z-axis in 3D geometric morphometrics based on sailfin silversides (Telmatherinidae).
This study investigated the impact of the third dimension in geometric morphometrics (GM) using sailfin silversides (Telmatherinidae) from the Malili Lakes of Sulawesi (Indonesia). The three morphospecies of the monophyletic "roundfin" radiation are laterally compressed and vary in shape traits. The results of 2D and 3D GM were compared and quantified to discuss the advantages and disadvantages of both methods for closely related species and their sexes. This approach focused on the head because it is far more complex and three-dimensionally structured than the trunk or the caudal region. The results revealed no significant benefit concerning repeatability and measurement error in 3D GM compared to 2D GM. The z-axis contributed substantially to the variance of the 3D data set but was irrelevant for discrimination of species and sexes in the approach. Limited gain in information was contrasted by substantially higher effort for 3D compared to the 2D analyses. The study concluded that 2D GM is the more efficient shape analysis approach for discriminating roundfins. Broader studies are needed to test which of the two methods is more efficient in distinguishing laterally compressed fishes in general. For future studies, due to the high investment required, this study recommends carefully evaluating the necessity of 3D GM. If in doubt, this study suggests testing for congruence between 2D and 3D GM with a subsample and consequently applying 2D GM in the case of high congruence.


Internally generated reactivation of single neurons in human hippocampus during free recall.
The emergence of memory, a trace of things past, into human consciousness is one of the greatest mysteries of the human mind. Whereas the neuronal basis of recognition memory can be probed experimentally in human and nonhuman primates, the study of free recall requires that the mind declare the occurrence of a recalled memory (an event intrinsic to the organism and invisible to an observer). Here, we report the activity of single neurons in the human hippocampus and surrounding areas when subjects first view cinematic episodes consisting of audiovisual sequences and again later when they freely recall these episodes. A subset of these neurons exhibited selective firing, which often persisted throughout and following specific episodes for as long as 12 seconds. Verbal reports of memories of these specific episodes at the time of free recall were preceded by selective reactivation of the same hippocampal and entorhinal cortex neurons. We suggest that this reactivation is an internally generated neuronal correlate for the subjective experience of spontaneous emergence of human recollection.


No detectable genetic correlation between male and female mating frequency in the stalk-eyed fly Cyrtodiopsis dalmanni.
There is much interest in explaining why female insects mate multiply. Females of the stalk-eyed fly Cyrtodiopsis dalmanni can mate several times each day in a lifetime which may span several months. There are many adaptive explanations, but one hypothesis that has received little rigorous empirical attention is that female multiple mating has evolved for non-adaptive reasons as a correlated response to selection for high male mating frequency rather than because of direct or indirect benefits accruing to females. We tested this hypothesis in stalk-eyed flies by measuring the mating frequency of females from lines that exhibited a direct response in males to artificial selection for increased ('high') and decreased ('low') male mating frequency. We found that the mating frequency of high-line females did not differ from that of low-line females. Hence, there was no support for a genetic correlation between male and female mating frequency in this species. Our study suggests that the genes which influence remating may not be the same in the sexes, and that females remate frequently in this species to gain as yet unidentified benefits.


High-throughput mouse phenomics for characterizing mammalian gene function.
We are entering a new era of mouse phenomics, driven by large-scale and economical generation of mouse mutants coupled with increasingly sophisticated and comprehensive phenotyping. These studies are generating large, multidimensional gene-phenotype data sets, which are shedding new light on the mammalian genome landscape and revealing many hitherto unknown features of mammalian gene function. Moreover, these phenome resources provide a wealth of disease models and can be integrated with human genomics data as a powerful approach for the interpretation of human genetic variation and its relationship to disease. In the future, the development of novel phenotyping platforms allied to improved computational approaches, including machine learning, for the analysis of phenotype data will continue to enhance our ability to develop a comprehensive and powerful model of mammalian gene-phenotype space.


Characterization of sulphate transporters in isolated bovine articular chondrocytes.
Uptake of SO(4) (2-) by articular chondrocytes is an essential step in the pathway for sulphation of glycosaminoglycans (GAGs), with mutations in SO(4) (2-) transport proteins resulting in abnormalities of skeletal growth. In the present study, the transporters mediating SO(4) (2-) transport in bovine articular chondrocytes have been characterized. Expression of candidate transporters was determined using RT-PCR, while SO(4) (2-) transport was measured in radioisotope flux experiments. RT-PCR experiments showed that bovine articular chondrocytes express three transporters known to transport SO(4) (2-): AE2 (SLC4a2), DTDST (SLC26a2), and SLC26a11. Other transporters--NaS-1 (SLC13a1), SAT-1 (SLC26a1), DRA (SLC26a3), SLC26a6 (PAT1), SLC26a7, SLC26a8 (Tat-1), and SLC26a9--were, however, not detected. In functional experiments, SO(4) (2-) uptake was temperature-sensitive, inhibited by 60% by DIDS (50 microM) and exhibited saturation kinetics, with a K(m) value of 16 mM. Uptake was also inhibited at alkaline extracellular pH. In further experiments, a K(i) value for DIDS inhibition of SO(4) (2-) efflux of 5 microM was recorded. A DIDS-sensitive component of SO(4) (2-) efflux persisted in solutions lacking Cl(-) ions. These data are interpreted as evidence for the preferential operation of carrier-mediated exchange of SO(4) (2-) for Cl(-), while an alternative SO(4) (2-)-OH(-) exchange mode is also possible.


There are no no-responders to low or high resistance training volumes among older women.
To assess the prevalence of non-responders to different tests and to compare the effects of different resistance training (RT) volumes on muscle strength, anthropometric and functional performance of older women. Three hundred seventy six women performed 12weeks of RT with either low or high volume (LV, 71.29±5.77years and HV 69.73±5.88years, respectively). Both groups performed the same exercises, and all parameters were held constant except for the number of sets performed per week. LV performed 8-12 for upper and 4-6 for lower body, while HV performed 16-20 and 8-10, respectively. Before and after the training period, the participants were tested for bench press and leg press 1RM, 30-s chair stand, 30-s arm curl, six-minute walk test, sit and reach, body weight and waist circumference. Both groups significantly improved in all strength and functional tests and reduced their body weight and waist circumference. ANOVA revealed higher gains in the leg press 1RM, 30-s arm curls and 6-min walk test for the HV group and higher increases in the results of the sit and reach test for the LV group. However, the differences were negligible and may be attributable to a type I error due to the large sample size. Non-responsiveness was not apparent in any subject, as a positive response on at least one outcome was present in every participant. Our results suggest that RT, even at low volume, improves waist circumference, muscle strength and physical function in the older population, with no evidence of non-responsiveness. Therefore, we should not be restrictive in prescribing this type of exercise to this population.


A next-generation sequencing method for gene doping detection that distinguishes low levels of plasmid DNA against a background of genomic DNA.
Gene doping confers health risks for athletes and is a threat to fair competition in sports. Therefore the anti-doping community has given attention on its detection. Previously published polymerase chain reaction-based methodologies for gene doping detection are targeting exon-exon junctions in the intron-less transgene. However, because these junctions are known, it would be relatively easy to evade detection by tampering with the copyDNA sequences. We have developed a targeted next-generation sequencing based assay for the detection of all exon-exon junctions of the potential doping genes, EPO, IGF1, IGF2, GH1, and GH2, which is resistant to tampering. Using this assay, all exon-exon junctions of copyDNA of doping genes could be detected with a sensitivity of 1296 copyDNA copies in 1000 ng of genomic DNA. In addition, promotor regions and plasmid-derived sequences are readily detectable in our sequence data. While we show the reliability of our method for a selection of genes, expanding the panel to detect other genes would be straightforward. As we were able to detect plasmid-derived sequences, we expect that genes with manipulated junctions, promotor regions, and plasmid or virus-derived sequences will also be readily detected.


Pathological Characterization Of IFNAR(-/-) Mice Infected With Bluetongue Virus Serotype 4.
Bluetongue virus (BTV) replicates in lymphoid tissues where infected mononuclear leukocytes secrete proinflammatory and vasoactive mediators that can contribute to bluetongue (BT) pathogenesis. Using the well-characterized IFNAR(-/-) mice animal model, we have now studied the histopathology and dynamics of leukocyte populations in different target tissues (spleen, thymus, and lung) during BTV-4 infection by histological and immunohistochemical techniques. The spleen and thymus of BTV-4 infected mice showed severe lymphoid depletion on H&E stained sections. This finding was confirmed by IHC, showing moderate decreased immunopositivity against CD3 in the thymus, and scarce immunoreactivity against CD3 and CD79 in the rest of the white pulp in the spleen, together with an increase in MAC387 immunostaining. BTV-4 infection also induced the expression of active caspase-3 in the spleen, where apoptotic debris was observed by H&E. A dramatic increase in iNOS immunoreactivity associated to necrotic areas of the white pulp was observed, being less noticeable in the thymus and the lung. The induction of pro-inflammatory cytokines in tissues where BTV replicates was evaluated by measuring transcript levels by RT-qPCR. BTV-4 infection led to enhance transcription of IFN-γ, TNF, IL-6, IL-12-p40, and IL-1β mRNA in the thymus, spleen and lung, correlating with the level of virus replication in these tissues. Disease progression and pathogenesis in IFNAR(-/-) mice closely mimics hallmarks of bluetongue disease in ruminants. IFNAR(-/-) mice are a good choice to facilitate a faster advance in the field of orbiviruses.


Eliminating complement factor D reduces photoreceptor susceptibility to light-induced damage.
Genetic risk factors such as variations in complement factors H (CFH) and B (CFB) have been implicated in the etiology of age-related macular degeneration. It has been hypothesized that inadequate control of complement-driven inflammation may be a major factor in disease pathogenesis. The authors tested the involvement of the complement system in an experimental model for oxidative stress-mediated photoreceptor degeneration, the light-damage mouse model. Changes in gene expression were assessed in BALB/c retinas in response to constant-light (CL) exposure using microarrays and real-time PCR. Susceptibility to CL exposure was tested in CFD(-/-) mice on a BALB/c background. Eyes were analyzed using electrophysiologic and histologic techniques. Genes encoding for proteins involved in complement activation were significantly upregulated after CL. The altered gene profiles were similar to proteins accumulated in drusen and to genes identified in the retina and RPE/choroid of patients with age-related macular degeneration. Cyclic-light reared CFD(-/-) and CFD(+/+) mice had indistinguishable rod function and number; however, after CL challenge, CFD(-/-) photoreceptors were significantly protected. These results suggest that rod degeneration in the CL-damaged retina involves the activity of the alternative complement pathway and that eliminating the alternative pathway is neuroprotective. Thus, the light damage albino mouse model may be a good model to study complement-mediated photoreceptor degeneration.


Increased neural correlations in primate auditory cortex during slow-wave sleep.
During sleep, changes in brain rhythms and neuromodulator levels in cortex modify the properties of individual neurons and the network as a whole. In principle, network-level interactions during sleep can be studied by observing covariation in spontaneous activity between neurons. Spontaneous activity, however, reflects only a portion of the effective functional connectivity that is activated by external and internal inputs (e.g., sensory stimulation, motor behavior, and mental activity), and it has been shown that neural responses are less correlated during external sensory stimulation than during spontaneous activity. Here, we took advantage of the unique property that the auditory cortex continues to respond to sounds during sleep and used external acoustic stimuli to activate cortical networks for studying neural interactions during sleep. We found that during slow-wave sleep (SWS), local (neuron-neuron) correlations are not reduced by acoustic stimulation remaining higher than in wakefulness and rapid eye movement sleep and remaining similar to spontaneous activity correlations. This high level of correlations during SWS complements previous work finding elevated global (local field potential-local field potential) correlations during sleep. Contrary to the prediction that slow oscillations in SWS would increase neural correlations during spontaneous activity, we found little change in neural correlations outside of periods of acoustic stimulation. Rather, these findings suggest that functional connections recruited in sound processing are modified during SWS and that slow rhythms, which in general are suppressed by sensory stimulation, are not the sole mechanism leading to elevated network correlations during sleep.


Targeting mesothelioma using an infectivity enhanced survivin-conditionally replicative adenoviruses.
Mesothelioma is a highly malignant neoplasm with no effective treatment. Conditionally replicative adenoviruses (CRAds) represent a promising new modality for the treatment of cancer in general. A key contribution in this regard is the introduction of tumor-selective viral replication for amplification of the initial inoculum in the neoplastic cell population. Under ideal conditions following cellular infection, the viruses replicate selectively in the infected tumor cells and kill the cells by cytolysis, leaving normal cells unaffected. However, to date there have been two limitations to clinical application of these CRAd agents; viral infectivity and tumor specificity have been poor. Herein we report on two CRAd agents, CRAd-S.RGD and CRAd-S.F5/3, in which the tumor specificity is regulated by a tumor-specific promoter, the survivin promoter, and the viral infectivity is enhanced by incorporating a capsid modification (RGD or F5/3) in the adenovirus fiber region. These CRAd agents effectively target human mesothelioma cell lines, induce strong cytoxicity in these cells in vitro, and viral replication in a H226 murine xenograft model in vivo. In addition, the survivin promoter has extremely low activity both in the non-transformed cell line, HMEC, and in human liver tissue. Our results suggest that the survivin-based CRAds are promising agents for targeting mesothelioma with low host toxicity. These agents should provide important insights into the identification of novel therapeutic strategies for mesothelioma.


Risk of ischemic stroke during the initiation period of α-blocker therapy among older men.
Alpha-blockers are notorious for their first-dose effect of acute hypotension during the early initiation period. Because acute cerebral hypoperfusion may precipitate an episode of ischemic stroke, we aimed to provide a quantitative estimate of the risk of ischemic stroke during the early initiation period of α-blocker therapy, using a self-controlled case series design. We identified all men aged 50 years or more as of 2007 who were incident users of α-blockers and had a diagnosis of ischemic stroke during the 2007-2009 study period using claims data from Taiwan's National Health Insurance claims database. The first day on which the α-blocker was prescribed was the index date. We partitioned different risk periods according to their relationship to the index date (pre-exposure risk periods 1 and 2 = ≤ 21 d and 22-60 d before index date, respectively; post-exposure risk periods 1 and 2 = ≤ 21 d and 22-60 d after index date, respectively); the remainder of the study period was defined as the unexposed period. We estimated the incidence rate ratio (IRR) of ischemic stroke in each risk period relative to the unexposed period using a conditional Poisson regression model. A total of 7502 men were included. Compared with the risk in the unexposed period, the risk of ischemic stroke was increased in post-exposure risk period 1 among all patients in the study population (adjusted IRR 1.40, 95% confidence interval [CI], 1.22-1.61) and among patients without concomitant prescriptions for other antihypertensive agents (adjusted IRR 2.11, 95% CI 1.73-2.57). Alpha-blocker therapy was associated with an increased risk of ischemic stroke during the early initiation period, especially among patients who were not taking other antihypertensive agents.


Differential caveolin-1 polarization in endothelial cells during migration in two and three dimensions.
Endothelial cell (EC) migration is a critical event during multiple physiological and pathological processes. ECs move in the plane of the endothelium to heal superficially injured blood vessels but migrate in three dimensions during angiogenesis. We herein investigate differences in these modes of movement focusing on caveolae and their defining protein caveolin-1. Using a novel approach for morphological analysis of transmigrating cells, we show that ECs exhibit a polarized distribution of caveolin-1 when traversing a filter pore. Strikingly, in these cells caveolin-1 seems to be released from caveolar structures in the cell rear and to relocalize at the cell front in a cytoplasmic form. In contrast, during planar movement caveolin-1 is concentrated at the rear of ECs, colocalizing with caveolae. The phosphorylatable Tyr14 residue of caveolin-1 is required for polarization of the protein during transmigration but does not alter polarization during planar movement. Palmitoylation of caveolin-1 is not essential for redistribution of the protein during either mode of movement. Thus, ECs migrating in three dimensions uniquely exhibit dissociation of caveolin-1 from caveolae and phosphorylation-dependent relocalization to the cell front.


Coffee Drinking and Mortality in 10 European Countries: A Multinational Cohort Study.
The relationship between coffee consumption and mortality in diverse European populations with variable coffee preparation methods is unclear. To examine whether coffee consumption is associated with all-cause and cause-specific mortality. Prospective cohort study. 10 European countries. 521 330 persons enrolled in EPIC (European Prospective Investigation into Cancer and Nutrition). Hazard ratios (HRs) and 95% CIs estimated using multivariable Cox proportional hazards models. The association of coffee consumption with serum biomarkers of liver function, inflammation, and metabolic health was evaluated in the EPIC Biomarkers subcohort (n = 14 800). During a mean follow-up of 16.4 years, 41 693 deaths occurred. Compared with nonconsumers, participants in the highest quartile of coffee consumption had statistically significantly lower all-cause mortality (men: HR, 0.88 [95% CI, 0.82 to 0.95]; P for trend < 0.001; women: HR, 0.93 [CI, 0.87 to 0.98]; P for trend = 0.009). Inverse associations were also observed for digestive disease mortality for men (HR, 0.41 [CI, 0.32 to 0.54]; P for trend < 0.001) and women (HR, 0.60 [CI, 0.46 to 0.78]; P for trend < 0.001). Among women, there was a statistically significant inverse association of coffee drinking with circulatory disease mortality (HR, 0.78 [CI, 0.68 to 0.90]; P for trend < 0.001) and cerebrovascular disease mortality (HR, 0.70 [CI, 0.55 to 0.90]; P for trend = 0.002) and a positive association with ovarian cancer mortality (HR, 1.31 [CI, 1.07 to 1.61]; P for trend = 0.015). In the EPIC Biomarkers subcohort, higher coffee consumption was associated with lower serum alkaline phosphatase; alanine aminotransferase; aspartate aminotransferase; γ-glutamyltransferase; and, in women, C-reactive protein, lipoprotein(a), and glycated hemoglobin levels. Reverse causality may have biased the findings; however, results did not differ after exclusion of participants who died within 8 years of baseline. Coffee-drinking habits were assessed only once. Coffee drinking was associated with reduced risk for death from various causes. This relationship did not vary by country. European Commission Directorate-General for Health and Consumers and International Agency for Research on Cancer.


Top-down pulses reduce prey population sizes and persistence.
Resource pulses are well documented and have important consequences for population dynamics relative to continuous inputs. However, pulses of top-down factors (e.g. predation) are less explored and appreciated in the ecological literature. Here, we use a simple differential equation population model to show how pulsed removals of individuals from a population alter population size relative to continuous dynamics. Pulsed removals result in lower equilibrium population sizes relative to continuous removals, and the differences are greatest at low population growth rates, high removal rates, and with large, infrequent pulses. Furthermore, the timing of the removal pulses (either stochastic or cyclic) affects population size. For example, cyclic removals are less likely than stochastic removals to result in population eradication, but when eradication occurs, the time until eradication is shorter for cyclic than with stochastic removals.


Metabolic Profiling as Well as Stable Isotope Assisted Metabolic and Proteomic Analysis of RAW 264.7 Macrophages Exposed to Ship Engine Aerosol Emissions: Different Effects of Heavy Fuel Oil and Refined Diesel Fuel.
Exposure to air pollution resulting from fossil fuel combustion has been linked to multiple short-term and long term health effects. In a previous study, exposure of lung epithelial cells to engine exhaust from heavy fuel oil (HFO) and diesel fuel (DF), two of the main fuels used in marine engines, led to an increased regulation of several pathways associated with adverse cellular effects, including pro-inflammatory pathways. In addition, DF exhaust exposure was shown to have a wider response on multiple cellular regulatory levels compared to HFO emissions, suggesting a potentially higher toxicity of DF emissions over HFO. In order to further understand these effects, as well as to validate these findings in another cell line, we investigated macrophages under the same conditions as a more inflammation-relevant model. An air-liquid interface aerosol exposure system was used to provide a more biologically relevant exposure system compared to submerged experiments, with cells exposed to either the complete aerosol (particle and gas phase), or the gas phase only (with particles filtered out). Data from cytotoxicity assays were integrated with metabolomics and proteomics analyses, including stable isotope-assisted metabolomics, in order to uncover pathways affected by combustion aerosol exposure in macrophages. Through this approach, we determined differing phenotypic effects associated with the different components of aerosol. The particle phase of diluted combustion aerosols was found to induce increased cell death in macrophages, while the gas phase was found more to affect the metabolic profile. In particular, a higher cytotoxicity of DF aerosol emission was observed in relation to the HFO aerosol. Furthermore, macrophage exposure to the gas phase of HFO leads to an induction of a pro-inflammatory metabolic and proteomic phenotype. These results validate the effects found in lung epithelial cells, confirming the role of inflammation and cellular stress in the response to combustion aerosols.


Ensemble cryo-EM uncovers inchworm-like translocation of a viral IRES through the ribosome.
Internal ribosome entry sites (IRESs) mediate cap-independent translation of viral mRNAs. Using electron cryo-microscopy of a single specimen, we present five ribosome structures formed with the Taura syndrome virus IRES and translocase eEF2•GTP bound with sordarin. The structures suggest a trajectory of IRES translocation, required for translation initiation, and provide an unprecedented view of eEF2 dynamics. The IRES rearranges from extended to bent to extended conformations. This inchworm-like movement is coupled with ribosomal inter-subunit rotation and 40S head swivel. eEF2, attached to the 60S subunit, slides along the rotating 40S subunit to enter the A site. Its diphthamide-bearing tip at domain IV separates the tRNA-mRNA-like pseudoknot I (PKI) of the IRES from the decoding center. This unlocks 40S domains, facilitating head swivel and biasing IRES translocation via hitherto-elusive intermediates with PKI captured between the A and P sites. The structures suggest missing links in our understanding of tRNA translocation.


The P gene product of hepatitis B virus is required as a structural component for genomic RNA encapsidation.
Encapsidation of the pregenomic RNA into nucleocapsids is a selective process which depends on specific RNA-protein interactions. The signal involved in the packaging of the hepatitis B virus (HBV) RNA pregenome was recently defined as a short sequence located near the 5' end of that molecule (Junker-Niepmann et al., EMBO J., in press), but it remained an open question which viral proteins are required. Using a genetic approach, we analyzed whether proteins derived from the HBV P gene play an important role in pregenome encapsidation. The results obtained with point mutations, deletions, and insertions scattered throughout the P gene clearly demonstrate that (i) a P gene product containing all functional domains is required both for the encapsidation of HBV pregenomic RNA and for packaging of nonviral RNAs fused to the HBV encapsidation signal, (ii) known enzymatic activities are not involved in the packaging reaction, suggesting that P protein is required as a structural component, and (iii) P protein acts primarily in cis, i.e., pregenomic RNAs from which P protein is synthesized are preferentially encapsidated.


Morphology of the male reproductive system of the social wasp, Polistes versicolor versicolor, with phylogenetic implications.
Variation in the morphology of the adult male reproductive system among different groups of Hymenoptera offer characteristics that help studies of behavior and the evolutionary history of this group. The objective of this study was to describe the adult male reproductive system of the wasp Polistes versicolor versicolor Olivier (Vespidae: Polistini). The reproductive systems were dissected, fixed and embedded for light microscopy. In P. v. versicolor, the reproductive system includes a pair of testes, each one with three fusiform follicles. From each follicle emerges an efferent duct that later join together, forming a deferent duct. The first half of the deferent duct is enlarged and differentiated into a region specialized for sperm storage, the seminal vesicle. At the post-vesicular region of each of the deferent ducts an accessory gland emerges. The seminal vesicle and the accessory gland are covered with a capsule forming a vesicle-gland complex, also observed in some species of North American Polistes. Sperm are released from testes in bundles, which are disorganized inside seminal vesicles. In the testicular follicles, 95 spermatozoa were observed per cyst on average.


Identification of nine novel mutations in the hepatocyte nuclear factor 1 alpha gene associated with maturity-onset diabetes of the young (MODY3).
Maturity-onset diabetes of the young (MODY) is a genetically heterogeneous subtype of non-insulin-dependent diabetes mellitus (NIDDM) characterised by early onset, autosomal dominant inheritance and a primary defect in insulin secretion. Recent studies have shown that mutations in the two functionally related transcription factors, hepatocyte nuclear factor 4 alpha (HNF-4alpha) and hepatocyte nuclear factor 1 alpha (HNF-1alpha) are associated with the MODY1 and MODY3 forms of diabetes respectively, whereas mutations in the enzyme glucokinase are the cause of the MODY2 form. We have examined 10 unrelated Caucasian families in which MODY/NIDDM co-segregated with markers for MODY3 for mutations in the HNF-1alpha gene (TCF1). Ten different mutations were observed in these families, all of which co-segregated with diabetes. There were no obvious relationships between the nature of the mutations observed (i.e. frameshift, nonsense, or missense) or their location in the gene with clinical features of diabetes (age at onset, severity) in these families. The mechanisms by which mutations in the HNF-1alpha gene cause diabetes mellitus are unclear but might include abnormal pancreatic islet development during foetal life thereby limiting their later function, as well as impaired transcriptional regulation of genes that play a key role in normal pancreatic beta cell function.


Platelet adhesion and aggregation under flow using microfluidic flow cells.
Platelet aggregation occurs in response to vascular injury where the extracellular matrix below the endothelium has been exposed. The platelet adhesion cascade takes place in the presence of shear flow, a factor not accounted for in conventional (static) well-plate assays. This article reports on a platelet-aggregation assay utilizing a microfluidic well-plate format to emulate physiological shear flow conditions. Extracellular proteins, collagen I or von Willebrand factor are deposited within the microfluidic channel using active perfusion with a pneumatic pump. The matrix proteins are then washed with buffer and blocked to prepare the microfluidic channel for platelet interactions. Whole blood labeled with fluorescent dye is perfused through the channel at various flow rates in order to achieve platelet activation and aggregation. Inhibitors of platelet aggregation can be added prior to the flow cell experiment to generate IC50 dose response data.


Effect of photonic crystal structure on the nonlinear optical anisotropy of birefringent porous silicon.
Anisotropic photonic crystal structures consisting of birefringent porous silicon layers with alternating porosity were fabricated. The in-plane birefringence formed as a result of anisotropic etching in Si(110) results in unique multilayered structures with two distinct photonic bandgaps for orthogonal light polarizations. Nonlinear optical studies based on the third-harmonic generation from these structures demonstrate variation in the symmetry of the nonlinear optical response.


Predicting the onset of knee pain: results from a 2-year prospective study of new workers.
To determine the relative contribution of work-related mechanical (injury) factors and psychosocial factors to the onset of a new episode of knee pain, in a cohort of newly employed workers. A prospective cohort study of newly employed workers from 12 diverse occupational settings in England (The New Workers Study). 859 newly employed workers, free of knee pain, were identified. Information about occupational mechanical factors (manual handling and postural activities), the occupational physical environment, and psychological and psychosocial factors was collected by self-completion questionnaires. Participants were followed up after 12 and 24 months to identify cases of knee pain onset. Generalised estimating equations were used to estimate the risk of new-onset knee pain, with respect to the exposures previously measured. In total, over the 2-year follow-up period, 108 cases of new-onset knee pain were observed. Mechanical load, postural factors, psychological distress and work-place psychosocial factors all influenced the risk of new-onset knee pain over the 2-year follow-up period. On multivariate analysis, two factors remained independently predictive of knee pain onset: lifting or carrying heavy weights in one hand, and the level of general psychological distress. In addition to mechanical (injury) factors, psychological factors are important risk factors for knee pain onset as shown in a population of young newly employed workers.


Autoimmunity in autism.
Autism spectrum disorders is a heterogenous group of neurodevelopmental disorders, the etiology or etiologies of which remain unknown. Increasing evidence of autoimmune phenomena in individuals with autism could represent the presence of altered or inappropriate immune responses in this disorder, and this immune system dysfunction may represent novel targets for treatment. Furthermore, in recent studies, antibodies directed against the fetal brain have been detected in some mothers of children with autism; these antibodies have the ability to alter behavioral outcomes in the offspring of animal models. A better understanding of the involvement of the immune response in early brain development, with respect to autism, may have important therapeutic implications.


Glycine transporter type 1 occupancy by bitopertin: a positron emission tomography study in healthy volunteers.
Deficient N-methyl-D-aspartate (NMDA) receptor transmission is thought to underlie schizophrenia. An approach for normalizing glutamate neurotransmission by enhancing NMDA receptor transmission is to increase glycine availability by inhibiting the glycine transporter type 1 (GlyT1). This study investigated the relationship between the plasma concentration of the glycine reuptake inhibitor bitopertin (RG1678) and brain GlyT1 occupancy. Healthy male volunteers received up to 175 mg bitopertin once daily, for 10-12 days. Three positron emission tomography scans, preceded by a single intravenous infusion of ∼30 mCi [(11)C]RO5013853, were performed: at baseline, on the last day of bitopertin treatment, and 2 days after drug discontinuation. Eighteen subjects were enrolled. At baseline, regional volume of distribution (V(T)) values were highest in the pons, thalamus, and cerebellum (1.7-2.7 ml/cm(3)) and lowest in cortical areas (∼0.8 ml/cm(3)). V(T) values were reduced to a homogeneous level following administration of 175 mg bitopertin. Occupancy values derived by a two-tissue five-parameter (2T5P) model, a simplified reference tissue model (SRTM), and a pseudoreference tissue model (PRTM) were overall comparable. At steady state, the relationship between bitopertin plasma concentration and GlyT1 occupancy derived by the 2T5P model, SRTM, and PRTM exhibited an EC(50) of ∼190, ∼200, and ∼130 ng/ml, respectively. E(max) was ∼92% independently of the model used. Bitopertin plasma concentration was a reliable predictor of occupancy because the concentration-occupancy relationship was superimposable at steady state and 2 days after drug discontinuation. These data allow understanding of the concentration-occupancy-efficacy relationship of bitopertin and support dose selection of future molecules.


Nitrogen uptake potential under different temperature-salinity conditions: Implications for nitrogen cycling under climate change scenarios.
As projected by climate change models, increase in sea surface temperature and precipitation in the future may alter nutrient cycling in the coastal regions due to potential changes in phytoplankton community structure and their ability to assimilate nitrogen (N) and carbon (C). An experiment simulating different temperature and salinity conditions (28°C-35 ambient conditions, 28ºC-31, 31ºC-35 and 31ºC-31) in mesocosms containing 1000 L of coastal water from the Arabian Sea was performed and N uptake rates were measured using 15N tracer technique on 2nd, 5th, 7th and 10th day of the experiment. The results show that, under all conditions, the total N (NO3- + NH4+) uptake rates were lower in the beginning and on the final day of the tracer experiment, while it peaked during middle, consistent with chlorophyll a concentrations. Total N uptake rate was significantly lower (p = 0.003) under ambient temperature-lower salinity condition (28ºC-31) than the others. This indicates that lowering of salinity in coastal regions due to excessive rainfall in the future may affect the N uptake potential of the phytoplankton, which may change the regional C and N budget.


A comparison of cardiac (31)P MRS at 1.5 and 3 T.
(31)P MRS was evaluated on normal volunteers at 1.5 and 3 T, and the signal-to-noise ratio (SNR) of the two field strengths was calculated. The in vivo spin-lattice, T(1), relaxation times for PCr and gamma-ATP, which are essential for correcting for the effects of radiofrequency saturation on the PCr/ATP ratio, were determined at 3 T. The T(1) values for six volunteers were 3.8 +/- 0.7 s for PCr (mean +/- SD) and 2.4 +/- 1.1 s for gamma-ATP, which are similar to reported values at 1.5 T, allowing us to use protocols developed at 1.5 T at the new clinical field strength of 3 T. Direct comparison between 1.5 T and 3 T in the same 10 subjects, using coils of identical geometry and identical pulse sequences gave a mean SNR for PCr at 3 T which was 206 +/- 94% of that at 1.5 T. The linewidth for PCr increased from 13 +/- 6 Hz at 1.5 T to 22 +/- 12 Hz at 3 T. The coefficient of variation in the measurement of PCr/ATP, based on the Cramer-Rao lower bounds, was reduced from 32 +/- 25% at 1.5 T to 18 +/- 13% at 3 T. Thus, (31)P MRS at 3 T is greatly improved by the increase in SNR compared with acquisitions at 1.5 T because of the higher field strength.


Eukaryotic translesion synthesis DNA polymerases: specificity of structure and function.
This review focuses on eukaryotic translesion synthesis (TLS) DNA polymerases, and the emphasis is on Saccharomyces cerevisiae and human Y-family polymerases (Pols) eta, iota, kappa, and Rev1, as well as on Polzeta, which is a member of the B-family polymerases. The fidelity, mismatch extension ability, and lesion bypass efficiencies of these different polymerases are examined and evaluated in the context of their structures. One major conclusion is that, despite the overall similarity of basic structural features among the Y-family polymerases, there is a high degree of specificity in their lesion bypass properties. Some are able to bypass a particular DNA lesion, whereas others are efficient at only the insertion step or the extension step of lesion bypass. This functional divergence is related to the differences in their structures. Polzeta is a highly specialized polymerase specifically adapted for extending primer termini opposite from a diverse array of DNA lesions, and depending upon the DNA lesion, it contributes to lesion bypass in a mutagenic or in an error-free manner. Proliferating cell nuclear antigen (PCNA) provides the central scaffold to which TLS polymerases bind for access to the replication ensemble stalled at a lesion site, and Rad6-Rad18-dependent protein ubiquitination is important for polymerase exchange.