PIP5K-driven PtdIns(4,5)P2 synthesis: regulation and cellular functions.
It has long been known that phosphoinositides are present in cellular membranes, but only in the past four decades has our understanding of their importance for proper cell function advanced significantly. Key to determining the biological roles of phosphoinositides is understanding the enzymes involved in their metabolism. Although many such enzymes have now been identified, there is still much to learn about their cellular functions. Phosphatidylinositol 4-phosphate 5-kinases (PIP5Ks) are a group of kinases that catalyse the production of phosphatidylinositol (4,5)-bisphosphate [PtdIns(4,5)P(2)]. As well as being a substrate for the enzymes phospholipase C (PLC) and phosphatidylinositol 3-kinase (PI3K), PtdIns(4,5)P(2) acts as a second messenger in its own right, influencing a variety of cellular processes. In this Commentary, we review how PIP5Ks are modulated to achieve regulated PtdIns(4,5)P(2) production, and discuss the role of these proteins in different cellular processes.


How we decide when a neonate needs a transfusion.
The decision to transfuse a neonate can be approached by addressing a series of questions that cover the cause of anaemia, alternatives to transfusion, the need for transfusion and the risks. Recent clinical trials of red cell transfusions have started to inform evidence-based transfusion practice, but have raised uncertainties about neurological outcomes when policies advocating use of fewer red cell transfusions at lower haemoglobin concentration (Hb) thresholds were tested. Red cell transfusions should be considered when the Hb <120 g/l for premature neonates requiring mechanical ventilation support, with lower thresholds applying for oxygen-dependent neonates not requiring ventilation or for late anaemia (Hb <70-100 g/l, depending on gestational and post-natal age). There is no recent high quality evidence to inform thresholds for prophylactic platelet transfusions in stable non-bleeding premature neonates with platelet count levels of 50 × 10(9) /l, although common practice has become more restrictive, using lower safe thresholds for platelet transfusion between 20 and 30 × 10(9) /l. A more appropriate transfusion strategy for fresh frozen plasma (FFP) in neonates is one that emphasizes the therapeutic use of FFP in the face of bleeding, rather than prophylactic use in stable non-bleeding neonates who often have mild to moderate apparent abnormalities of standard coagulation tests, after allowing for appropriate reference ranges.


Smoking cessation in workplace settings: quit rates and determinants in a group behaviour therapy programme.
To capitalise on the opportunities that the smoking ban legislation in Switzerland offers for the prevention of tobacco-related diseases, a smoking cessation programme in a workplace setting was developed and implemented in companies across the language and cultural regions of Switzerland. Our goal was to identify factors associated with relapse into smoking that may be overcome during training sessions or that should be considered for the optimisation of future interventions. Between 2006 and 2012, 1287 smokers aged 16 to 68 years voluntarily attended smoking cessation training at their workplace. The intervention was based on a cognitive behavioural group therapy combined with individual proactive telephone counselling. The evaluation consisted of three anonymised questionnaires (pre- and postintervention, and 12-month follow-up). In this prospective cohort study, we investigated the association of smoking quit rates with training and participant characteristics, including withdrawal symptoms, by use of multilevel logistic regression analysis with a random intercept for training courses. The self-reported abstinence rate was 72.4% at the end of the training, and 18.6% 1 year later. The risk of relapse during the training was positively associated with the number of years and daily cigarettes smoked, and negatively with increased appetite, sleeping troubles and satisfaction with learned techniques. Failed abstinence within the first year was associated with younger age, higher numbers of daily smoked cigarettes and unsuccessful recent quit attempts. Our evaluation suggests that younger and more addicted smokers attending smoking cessation trainings may need additional support to achieve long lasting abstinence rates. Offering smoking cessation training in a workplace setting can achieve reasonable long-term quit rates, but a subset of employees needs additional support at the group or personal level. Group behaviour therapy could be an effective method to achieve long-term smoking abstinence. The workplace is an important setting to reach and encourage a large number of smokers to participate in a smoking cessation programme, but only few studies investigated its potential. The findings of this study of a modern real-life workplace-based smoking cessation programme endorse the effectiveness and viability of cognitive behavioural group therapy. This group-level intervention at the workplace may be insufficient for young and heavy smokers, as well as women with increased appetite after cessation, who potentially benefit from re-intervention and additional individual level counselling.


An Eco1-independent sister chromatid cohesion establishment pathway in S. cerevisiae.
Cohesion between sister chromatids, mediated by the chromosomal cohesin complex, is a prerequisite for their alignment on the spindle apparatus and segregation in mitosis. Budding yeast cohesin first associates with chromosomes in G1. Then, during DNA replication in S-phase, the replication fork-associated acetyltransferase Eco1 acetylates the cohesin subunit Smc3 to make cohesin's DNA binding resistant to destabilization by the Wapl protein. Whether stabilization of cohesin molecules that happen to link sister chromatids is sufficient to build sister chromatid cohesion, or whether additional reactions are required to establish these links, is not known. In addition to Eco1, several other factors contribute to cohesion establishment, including Ctf4, Ctf18, Tof1, Csm3, Chl1 and Mrc1, but little is known about their roles. Here, we show that each of these factors facilitates cohesin acetylation. Moreover, the absence of Ctf4 and Chl1, but not of the other factors, causes a synthetic growth defect in cells lacking Eco1. Distinct from acetylation defects, sister chromatid cohesion in ctf4Δ and chl1Δ cells is not improved by removing Wapl. Unlike previously thought, we do not find evidence for a role of Ctf4 and Chl1 in Okazaki fragment processing, or of Okazaki fragment processing in sister chromatid cohesion. Thus, Ctf4 and Chl1 delineate an additional acetylation-independent pathway that might hold important clues as to the mechanism of sister chromatid cohesion establishment.


Longitudinal changes in COVID-19 vaccination intent among South African adults: evidence from the NIDS-CRAM panel survey, February to May 2021.
COVID-19 vaccine hesitancy has threatened the ability of many countries worldwide to contain the pandemic. Given the severe impact of the pandemic in South Africa and disruptions to the roll-out of the vaccine in early 2021, slower-than-expected uptake is a pressing public health challenge in the country. We examined longitudinal changes in COVID-19 vaccination intent among South African adults, as well as determinants of intent to receive a vaccine. We used longitudinal data from Wave 4 (February/March 2021) and Wave 5 (April/May 2021) of the National Income Dynamics Study: Coronavirus Rapid Mobile Survey (NIDS-CRAM), a national and broadly representative panel survey of adults in South Africa. We conducted cross-sectional analyses on aggregate and between-group variation in vaccination intent, examined individual-level changes between waves, and modeled demographic predictors of intent. We analysed data for 5629 (Wave 4; 48% male, mean age 41.5 years) and 5862 (Wave 5; 48% male, mean age 41.6 years) respondents. Willingness to get a COVID-19 vaccine significantly increased from 70.8% (95% CI: 68.5-73.1) in Wave 4 to 76.1% (95% CI: 74.2-77.8) in Wave 5. Individual-level analyses indicated that only 6.6% of respondents remained strongly hesitant between survey waves. Although respondents aged 18-24 years were 8.5 percentage points more likely to report hesitancy, hesitant respondents in this group were 5.6 percentage points more likely to change their minds by Wave 5. Concerns about rushed testing and safety of the vaccines were frequent and strongly-held reasons for hesitancy. Willingness to receive a COVID-19 vaccine has increased among adults in South Africa, and those who were entrenched in their reluctance make up a small proportion of the country's population. Younger adults, those in formal housing, and those who trusted COVID-19 information on social media were more likely to be hesitant. Given that stated vaccination intent may not translate into behaviour, our finding that three-quarters of the population were willing to accept the vaccine may reflect an upper bound. Vaccination promotion campaigns should continue to frame vaccine acceptance as the norm and tailor strategies to different demographic groups.


Hepatic insulin signaling is required for obesity-dependent expression of SREBP-1c mRNA but not for feeding-dependent expression.
Dissecting the role of insulin in the complex regulation of triglyceride metabolism is necessary for understanding dyslipidemia and steatosis. Liver insulin receptor knockout (LIRKO) mice show that in the physiological context of feeding, hepatic insulin signaling is not required for the induction of mTORC1, an upstream activator of the lipogenic regulator, SREBP-1c. Feeding induces SREBP-1c mRNA in LIRKO livers, though not to the extent observed in controls. A high fructose diet also partially induces SREBP-1c and lipogenic gene expression in LIRKO livers. Insulin signaling becomes more important in the pathological context of obesity, as knockdown of the insulin receptor in ob/ob mice, a model of Type 2 diabetes, using antisense oligonucleotides, abolishes the induction of SREBP-1c and its targets by obesity and ameliorates steatosis. Thus, insulin-independent signaling pathways can partially compensate for insulin in the induction of SREBP-1c by feeding but the further induction by obesity/Type 2 diabetes is entirely dependent upon insulin.


CT and MRI findings in X-linked progressive deafness.
Congenital X-linked mixed deafness is a rare anomaly that has typical features and can be diagnosed on the basis of progressive mixed hearing loss and the typical imaging findings. Recognition of these findings may alter the course of treatment and perilymph gushing can be avoided. A 10-year-old male patient presented with a history of progressive hearing loss. Computed tomography of the temporal bones showed bulbous dilatation of the fundi of the internal auditory canals (IAC) and the absence of the bony plates separating the basal turn of the cochleas and IAC. Magnetic resonance imaging demonstrated obliteration of the labyrinthine spaces at the right side.


Convergent development of anodic bacterial communities in microbial fuel cells.
Microbial fuel cells (MFCs) are often inoculated from a single wastewater source. The extent that the inoculum affects community development or power production is unknown. The stable anodic microbial communities in MFCs were examined using three inocula: a wastewater treatment plant sample known to produce consistent power densities, a second wastewater treatment plant sample, and an anaerobic bog sediment. The bog-inoculated MFCs initially produced higher power densities than the wastewater-inoculated MFCs, but after 20 cycles all MFCs on average converged to similar voltages (470±20 mV) and maximum power densities (590±170 mW m(-2)). The power output from replicate bog-inoculated MFCs was not significantly different, but one wastewater-inoculated MFC (UAJA3 (UAJA, University Area Joint Authority Wastewater Treatment Plant)) produced substantially less power. Denaturing gradient gel electrophoresis profiling showed a stable exoelectrogenic biofilm community in all samples after 11 cycles. After 16 cycles the predominance of Geobacter spp. in anode communities was identified using 16S rRNA gene clone libraries (58±10%), fluorescent in-situ hybridization (FISH) (63±6%) and pyrosequencing (81±4%). While the clone library analysis for the underperforming UAJA3 had a significantly lower percentage of Geobacter spp. sequences (36%), suggesting that a predominance of this microbe was needed for convergent power densities, the lower percentage of this species was not verified by FISH or pyrosequencing analyses. These results show that the predominance of Geobacter spp. in acetate-fed systems was consistent with good MFC performance and independent of the inoculum source.


Structural insights into the stabilization of MALAT1 noncoding RNA by a bipartite triple helix.
Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is a highly abundant nuclear long noncoding RNA that promotes malignancy. A 3'-stem-loop structure is predicted to confer stability by engaging a downstream A-rich tract in a triple helix, similar to the expression and nuclear retention element (ENE) from the KSHV polyadenylated nuclear RNA. The 3.1-Å-resolution crystal structure of the human MALAT1 ENE and A-rich tract reveals a bipartite triple helix containing stacks of five and four U•A-U triples separated by a C+•G-C triplet and C-G doublet, extended by two A-minor interactions. In vivo decay assays indicate that this blunt-ended triple helix, with the 3' nucleotide in a U•A-U triple, inhibits rapid nuclear RNA decay. Interruption of the triple helix by the C-G doublet induces a 'helical reset' that explains why triple-helical stacks longer than six do not occur in nature.


Differential effects of estrogen and progesterone on development of primate secondary follicles in a steroid-depleted milieu in vitro.
What are the direct effects of progesterone (P4) and estradiol (E2) on the development and function of primate follicles in vitro from the pre-antral to early antral stage? In a steroid-depleted milieu, E2 improved follicle survival, growth, antrum formation and oocyte health, whereas P4 exerted minimal beneficial effects on follicle survival and reduced oocyte health. Effects of P4 and E2 on follicle development have been studied primarily in large antral and pre-ovulatory follicles. Chronic P4 exposure suppresses antral follicle growth, but acute P4 exposure promotes oocyte maturation in pre-ovulatory follicles. Effects of E2 can be stimulatory or inhibitory depending upon species, dose and duration of exposure. Non-human primate model, randomized, control versus treatment. Macaque (n = 6) secondary follicles (n = 24 per animal per treatment group) were cultured for 5 weeks. Adult rhesus macaque secondary follicles were encapsulated in 0.25% alginate and cultured individually in media containing follicle stimulating hormone plus (i) vehicle, (ii) a steroid-synthesis inhibitor, trilostane (TRL, 250 ng/ml), (iii) TRL + low E2 (100 pg/ml) or progestin (P, 10 ng/ml R5020) and (iv) TRL + high E2 (1 ng/ml E2) or P (100 ng/ml R5020). Follicles reaching the antral stage (≥750 µm) were treated with human chorionic gonadotrophin for 34 h. End-points included follicle survival, antrum formation, growth pattern, plus oocyte health and maturation status, as well as media concentrations of P4, E2 and anti-Müllerian hormone (AMH). In a steroid-depleted milieu, low dose, but not high dose, P improved (P < 0.05) follicle survival, but had no effect (P > 0.05) on antrum formation and AMH production. Low-dose P increased (P < 0.05) P4 production in fast-grow follicles, and both doses of P elevated (P < 0.05) E2 production in slow-grow follicles. Additionally, low-dose P increased (P < 0.05) the percentage of no-grow follicles, and high-dose P promoted oocyte degeneration. In contrast, E2, in a steroid-depleted milieu, improved (P < 0.05) follicle survival, growth, antrum formation and oocyte health. E2 had no effect on P4 or E2 production. Follicles exposed to E2 yielded mature oocytes capable of fertilization and early cleavage, at a rate similar to untreated control follicles. This study is limited to in vitro effects of P and E2 during the interval from the secondary to small antral stage of macaque follicles. This study provides novel information on the direct actions of P4 and E2 on primate pre-antral follicle development. Combined with our previous report on the actions of androgens, our findings suggest that androgens appear to be a survival factor but hinder antral follicle differentiation, E2 appears to be a survival and growth factor at the pre-antral and early antral stage, whereas P4 may not be essential during early folliculogenesis in primates. NIH P50 HD071836 (NCTRI), NIH ORWH/NICHD 2K12HD043488 (BIRCWH), ONPRC 8P51OD011092. There are no conflicts of interest.


The use of arm span as a predictor of height: A study of South Indian women.
This study was undertaken to find out the relationship between various body parameters to identify the measurement that correlates most closely to stature. Sitting height, standing height, arm span and leg lengths of 505 healthy women between the ages of 20 and 29 were measured. The relationship between the parameters was determined by computing correlation coefficients and 95% confidence intervals. The correlation between sitting height, arm span and leg length were poor (r=0.56 and 0.29 respectively), while the correlation of arm span and leg length with standing height were good (r=0.82 and 0.84 respectively). These estimations can be of use in quantifying the age related loss in stature and in identifying individuals with disproportionate growth abnormalities and skeletal dysplasias. It can also be useful to find out the alterations in the height of an individual that may occur due to progressive deformities of the spine. The adequacy of the correction of spinal deformities may also be facilitated by computing the ratio of arm span to standing height.


Analytic calculation of finite-population reproductive numbers for direct- and vector-transmitted diseases with homogeneous mixing.
The basic reproductive number, R0, provides a foundation for evaluating how various factors affect the incidence of infectious diseases. Recently, it has been suggested that, particularly for vector-transmitted diseases, R0 should be modified to account for the effects of finite host population within a single disease transmission generation. Here, we use a transmission factor approach to calculate such "finite-population reproductive numbers," under the assumption of homogeneous mixing, for both vector-borne and directly transmitted diseases. In the case of vector-borne diseases, we estimate finite-population reproductive numbers for both host-to-host and vector-to-vector generations, assuming that the vector population is effectively infinite. We find simple, interpretable formulas for all three of these quantities. In the direct case, we find that finite-population reproductive numbers diverge from R0 before R0 reaches half of the population size. In the vector-transmitted case, we find that the host-to-host number diverges at even lower values of R0, while the vector-to-vector number diverges very little over realistic parameter ranges.


Cycloheximide-Producing Streptomyces Associated With Xyleborinus saxesenii and Xyleborus affinis Fungus-Farming Ambrosia Beetles.
Symbiotic microbes help a myriad of insects acquire nutrients. Recent work suggests that insects also frequently associate with actinobacterial symbionts that produce molecules to help defend against parasites and predators. Here we explore a potential association between Actinobacteria and two species of fungus-farming ambrosia beetles, Xyleborinus saxesenii and Xyleborus affinis. We isolated and identified actinobacterial and fungal symbionts from laboratory reared nests, and characterized small molecules produced by the putative actinobacterial symbionts. One 16S rRNA phylotype of Streptomyces (XylebKG-1) was abundantly and consistently isolated from the galleries and adults of X. saxesenii and X. affinis nests. In addition to Raffaelea sulphurea, the symbiont that X. saxesenii cultivates, we also repeatedly isolated a strain of Nectria sp. that is an antagonist of this mutualism. Inhibition bioassays between Streptomyces griseus XylebKG-1 and the fungal symbionts from X. saxesenii revealed strong inhibitory activity of the actinobacterium toward the fungal antagonist Nectria sp. but not the fungal mutualist R. sulphurea. Bioassay guided HPLC fractionation of S. griseus XylebKG-1 culture extracts, followed by NMR and mass spectrometry, identified cycloheximide as the compound responsible for the observed growth inhibition. A biosynthetic gene cluster putatively encoding cycloheximide was also identified in S. griseus XylebKG-1. The consistent isolation of a single 16S phylotype of Streptomyces from two species of ambrosia beetles, and our finding that a representative isolate of this phylotype produces cycloheximide, which inhibits a parasite of the system but not the cultivated fungus, suggests that these actinobacteria may play defensive roles within these systems.


Standards for Pediatric Immunization Practices: an osteopathic physician's commentary and 'reality check'.
The "Standards for Pediatric Immunization Practices," recommended by the National Vaccine Advisory Committee and approved by the US Public Health Service, provides guidance to clinicians as to what the National Vaccine Advisory Committee considers optimal practice. Each of the 18 recommendations reflects the ideal; unfortunately, life in the trenches is less than ideal. Providers must reflect on each point and consider how they might modify their own practices to help us to realize the goal of Healthy People 2000, that is, 90% immunization coverage by the child's second birthday. The Standards are clear. They are reprinted herein from MMWR (April 23, 1993;42[5]). Following each is a response, sort of a reality check, from the author, a practicing clinician and practice administrator.


Impact of a statewide Internet-based tobacco cessation intervention.
An increasing number of people have access to the Internet, and more people are seeking tobacco cessation resources online every year. Despite the proliferation of various online interventions and their evident acceptance and reach, little research has addressed their impact in the real world. Typically, low response rates to Internet-based follow-up surveys generate unrepresentative samples and large confidence intervals when reporting results. The aim of this study was to achieve a high response rate on follow-up evaluation in order to better determine the impact of an Internet-based tobacco cessation intervention provided to tobacco users in Minnesota, United States. Participants included 607 men and women aged 18 and over residing in Minnesota who self-reported current tobacco use when registering for an Internet-based tobacco cessation program between February 2 and April 13, 2004. Participants were given access to an interactive website with features including social support, expert systems, proactive email, chat sessions, and online counselors. Mixed-mode follow-up (online survey with telephone survey for online nonrespondents) occurred 6 months after registration. Of the study participants, 77.6% (471/607) responded to the 6-month follow-up survey (39.4% online and 38.2% by telephone). Among respondents, 17.0% (80/471, 95% CI = 13.6%-20.4%) reported that they had not smoked in the past 7 days (observed rate). Assuming all nonrespondents were still smoking (missing=smoking rate), the quit rate was 13.2% (80/607, 95% CI = 10.5%-15.9%). This mixed-mode follow-up survey of an online smoking cessation program achieved a high response rate and provides a more accurate estimate of long-term cessation rates than has been previously reported. Quit rates for the Internet-based tobacco cessation program were higher than those expected for unassisted quit attempts and are comparable to other evidence-based behavioral interventions. The similarities between quit rates demonstrates that an Internet-based cessation program may have as great an impact as, and can have wider reach than, other cessation programs such as those delivered by telephone. With over 100000 people having visited the website and over 23000 having registered, a 6-month self-reported quit rate of 13.2% suggests that the quitplan.com program helped over 3000 Minnesotans remain tobacco free for at least 6 months. Results of this study suggest that an Internet-based cessation program is a useful tool in states' efforts to provide comprehensive cessation tools for smokers.


Live-cell monochromatic dual-label sub-diffraction microscopy by mt-pcSOFI.
We expand photochromic super-resolution optical fluctuation imaging (pcSOFI) to monochromatic dual-channel sub-diffraction microscopy. Multi-tau (mt-)pcSOFI unmixes spectrally identical reversibly switchable fluorescent proteins (RSFPs) based on their blinking kinetics. We show that mt-pcSOFI can be used to simultaneously image two structures in living cells with existing RSFPs and the newly developed ffDronpa-F.


The Toxoplasma gondii Active Serine Hydrolase 4 Regulates Parasite Division and Intravacuolar Parasite Architecture.
Hydrolase are enzymes that regulate diverse biological processes, including posttranslational protein modifications. Recent work identified four active serine hydrolases (ASHs) in Toxoplasma gondii as candidate depalmitoylases. However, only TgPPT1 (ASH1) has been confirmed to remove palmitate from proteins. ASH4 (TgME49_264290) was reported to be refractory to genetic disruption. We demonstrate that recombinant ASH4 is an esterase that processes short acyl esters but not palmitoyl thioesters. Genetic disruption of ASH4 causes defects in cell division and premature scission of parasites from residual bodies. These defects lead to the presence of vacuoles with a disordered intravacuolar architecture, with parasites arranged in pairs around multiple residual bodies. Importantly, we found that the deletion of ASH4 correlates with a defect in radial dispersion from host cells after egress. This defect in dispersion of parasites is a general phenomenon that is observed for disordered vacuoles that occur at low frequency in wild-type parasites, suggesting a possible general link between intravacuolar organization and dispersion after egress.IMPORTANCE This work defines the function of an enzyme in the obligate intracellular parasite Toxoplasma gondii We show that this previously uncharacterized enzyme is critical for aspects of cellular division by the parasite and that loss of this enzyme leads to parasites with cell division defects and which also are disorganized inside their vacuoles. This leads to defects in the ability of the parasite to disseminate from the site of an infection and may have a significant impact on the parasite's overall infectivity of a host organism.


Evaluation of the chicken transcriptome by SAGE of B cells and the DT40 cell line.
The understanding of whole genome sequences in higher eukaryotes depends to a large degree on the reliable definition of transcription units including exon/intron structures, translated open reading frames (ORFs) and flanking untranslated regions. The best currently available chicken transcript catalog is the Ensembl build based on the mappings of a relatively small number of full length cDNAs and ESTs to the genome as well as genome sequence derived in silico gene predictions. We use Long Serial Analysis of Gene Expression (LongSAGE) in bursal lymphocytes and the DT40 cell line to verify the quality and completeness of the annotated transcripts. 53.6% of the more than 38,000 unique SAGE tags (unitags) match to full length bursal cDNAs, the Ensembl transcript build or the genome sequence. The majority of all matching unitags show single matches to the genome, but no matches to the genome derived Ensembl transcript build. Nevertheless, most of these tags map close to the 3' boundaries of annotated Ensembl transcripts. These results suggests that rather few genes are missing in the current Ensembl chicken transcript build, but that the 3' ends of many transcripts may not have been accurately predicted. The tags with no match in the transcript sequences can now be used to improve gene predictions, pinpoint the genomic location of entirely missed transcripts and optimize the accuracy of gene finder software.


Interactions of saponin with microsomal membranes isolated from vascular smooth muscle.
The effects of the plant glycosides extract, saponin, on various biochemical properties of microsomal fractions isolated from dog aortae and mesenteric arteries were investigated. These properties include binding and transport of Ca2+, ouabain-sensitive K+ activated p-nitrophenyl phosphatase (hereafter termed K+-pNPPase) and binding of radiolabelled antagonists to adrenoceptors. We found that saponin inhibited both Ca2+ binding and transport by the vascular muscle microsomes in a dose-dependent manner, but it caused marked enhancement of the K+-pNPPase activity. Saponin only slightly reduced the Bmax, but not the Kd of [125I]-iodocyanopindolol binding to the beta-adrenoceptors of the dog mesenteric artery microsomes. A similar finding on the [3H]-prazosin binding to the alpha 1-adrenoceptor in dog aortic microsomes was also observed. Our findings are consistent with the "skinning" action of saponin on the smooth muscle and provide direct evidence that saponin does not attenuate the recognition properties of the adrenoceptors in the cell membranes.


ONS Guidelines™ for Cancer Treatment-Related Hot Flashes in Women With Breast Cancer and Men With Prostate Cancer.
Hot flashes are a common and troublesome side effect of surgery or endocrine therapy. They may lead to physical and psychological distress and negatively affect quality of life. This clinical practice guideline presents evidence-based recommendations for pharmacologic, behavioral, and natural health product interventions for treatment-related hot flashes in patients with breast or prostate cancer. An interprofessional panel of healthcare professionals with patient representation prioritized clinical questions and patient outcomes for the management of hot flashes. Systematic reviews of the literature were conducted. The GRADE (Grading of Recommendations Assessment, Development, and Evaluation) approach was used to assess the evidence and make recommendations. The panel agreed on 14 pharmacologic, behavioral, and natural health recommendations. Conditional recommendations include the use of antidepressants rather than no treatment, physical activity rather than no treatment, and the avoidance of gabapentin and dietary supplements in the treatment of hot flashes. //onf.ons.org/ons-guidelines-hot-flashes-supplementary-material.


Geranylgeraniol suppresses the viability of human DU145 prostate carcinoma cells and the level of HMG CoA reductase.
The rate-limiting enzyme of the mevalonate pathway, 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, provides essential intermediates for the prenylation of nuclear lamins and Ras and dolichol-mediated glycosylation of growth factor receptors. The diterpene geranylgeraniol downregulates the level of HMG CoA reductase and suppresses the growth of human liver, lung, ovary, pancreas, colon, stomach, and blood tumors. We evaluated the growth-suppressive activity of geranylgeraniol in human prostate carcinoma cells. Geranylgeraniol induced dose-dependent suppression of the viability of human DU145 prostate carcinoma cells (IC50=80±18 µmol/L, n=5) following 72-h incubations in 96-well plates. Cell cycle was arrested at the G1 phase with a concomitant decrease in cyclin D1 protein. Geranylgeraniol-induced apoptosis was detected by flow cytometric analysis, fluorescence microscopy following acridine orange and ethidium bromide dual staining, and caspase-3 activation. Geranylgeraniol-induced viability suppression was accompanied by concentration-dependent decrease in the level of HMG CoA reductase protein. As a nonsterol molecule that downregulates HMG CoA reductase in the presence of sterols, geranylgeraniol may have potential in the chemoprevention and/or therapy of human prostate cancer.


Sex-linked inheritance of hearing and song in the Belgian Waterslager canary.
Belgian Waterslager canaries have less sensitive hearing at high frequencies and produce songs with more energy at low frequencies than wild-type canaries. A backcross pedigree between Belgian Waterslager canaries and a domestic strain with wild-type song revealed inheritance patterns consistent with a factor of major effect located on the Z sex chromosome affecting both poor high-frequency hearing at 4 kHz and the relative energy in the spectra of the learned songs of males. Hearing thresholds at 4 kHz were significant predictors of the relative amount of song energy at 4 kHz for individual males. One hypothesis for the mechanistic basis of this correlation between hearing and song abnormalities is that a reduction in the ability to hear higher-frequency songs biases males towards learning lower-frequency songs.


Peptidylarginine Deiminase Inhibition Prevents Diabetes Development in NOD Mice.
Protein citrullination plays a role in several autoimmune diseases. Its involvement in murine and human type 1 diabetes has recently been recognized through the discovery of antibodies and T-cell reactivity against citrullinated peptides. In the current study, we demonstrate that systemic inhibition of peptidylarginine deiminases (PADs), the enzymes mediating citrullination, through BB-Cl-amidine treatment, prevents diabetes development in NOD mice. This prevention was associated with reduced levels of citrullination in the pancreas, decreased circulating autoantibody titers against citrullinated glucose-regulated protein 78, and reduced spontaneous neutrophil extracellular trap formation of bone marrow-derived neutrophils. Moreover, BB-Cl-amidine treatment induced a shift from Th1 to Th2 cytokines in the serum and an increase in the frequency of regulatory T cells in the blood and spleen. In the pancreas, BB-Cl-amidine treatment preserved insulin production and was associated with a less destructive immune infiltrate characterized by reduced frequencies of effector memory CD4+ T cells and a modest reduction in the frequency of interferon-γ-producing CD4+ and CD8+ T cells. Our results point to a role of citrullination in the pathogenesis of autoimmune diabetes, with PAD inhibition leading to disease prevention through modulation of immune pathways. These findings provide insight in the potential of PAD inhibition for treating autoimmune diseases like type 1 diabetes.


Lamin proteolysis facilitates nuclear events during apoptosis.
Expression of the adenovirus E1A oncogene stimulates both cell proliferation and p53-dependent apoptosis in rodent cells. p53 implements apoptosis in all or in part through transcriptional activation of bax, the product of which promotes cell death. The adenovirus E1B 19K product is homologous in sequence and in function to Bcl-2, both of which bind to and inhibit the activity of Bax and thereby suppress apoptosis. The E1B 19K protein also interacts with the nuclear lamins, but the role of this interaction in the regulation of apoptosis is not known. Lamins are, however, substrates for members of the interleukin-1 beta-converting enzyme (ICE) family of cysteine proteases that are activated during apoptosis and function downstream of Bcl-2 in the cell death pathway. lamins are degraded during E1A-induced p53-dependent apoptosis. Lamin A and C are cleaved into 47- and 37-kD fragments, respectively, and the site of proteolysis is mapped to a conserved aspartic acid residue at position 230. The cleavage of lamins during apoptosis is consistent with the activation of an ICE-related cysteine protease down-stream of p53. No lamin protease activity was detected in cells expressing the E1B 19K protein, indicating that 19K functions upstream of protease activation in inhibiting apoptosis. Substitution of the aspartic acid at the cleavage site produced a mutant lamin protein that was resistant to proteolysis both in vitro and in vivo. Expression of uncleavable mutant lamin A or B attenuated apoptosis, delaying cell death and the associated DNA fragmentation by 12 h. Mutant lamin expressing cells failed to show the signs of chromatin condensation and nuclear shrinkage typical of cell death by apoptosis. Instead, the nuclear envelope collapsed and the nuclear lamina remained intact. However, the late stage of apoptosis was morphologically unaltered and formation of apoptotic bodies was evident. Thus, lamin breakdown by proteolytic degradation facilitates the nuclear events of apoptosis perhaps by facilitating nuclear breakdown.


Modulation of CYP1A1, CYP1A2 and CYP1B1 expression by cabbage juices and indoles in human breast cell lines.
Epidemiological studies have shown that consumption of cabbage and sauerkraut is connected with significant reduction of breast cancer incidences. Estrogens are considered a major breast cancer risk factor and their metabolism by P450 enzymes substantially contributes to carcinogenic activity. The aim of this study was to investigate the effect of cabbage and sauerkraut juices of different origin on the expression profile of the estrogen metabolism key enzymes (CYP1A1, CYP1A2, CYP1B1) in breast cell lines MCF7, MDA-MB-231, and MCF10A. The effects of cabbage juices were compared with that exerted by indole-3-carbinol (I3C) and 3,3'-diindolylmethane (DIM). The treatment with cabbage juices or indoles for 72 h affected the expression of CYP1 family genes in cell-type dependent manner. Their induction was found in all cell lines, but the ratio of CYP1A1 to CYP1B1 was 1.22- to 10.6-fold in favor to CYP1A1 in MCF7 and MCF10A cells. Increased levels of CYP1A2 in comparison with CYP1B1 were also observed in MCF7 cells. In contrast, in MDA-MB-231 cells CYP1B1 was preferentially induced. Since the cell lines investigated differ in invasion capacity, these results support epidemiological observations and partly explain the mechanism of the chemopreventive activity of white cabbage products.


Maternal occupational exposure and congenital malformations.
A case-referent study was conducted to assess the risk of congenital malformations in relation to maternal occupational exposure before and during pregnancy. Three hundred and twenty-five cases of major malformations and 325 normal (at birth) referents identified in 15 maternity hospitals were included in the study. The occupational history obtained from an interview of the mother was blindly reviewed by an industrial hygienist who assessed the presence of chemical exposure and the probability of exposure. The results suggested that mothers of the case children with oral clefts were more often exposed to solvents during pregnancy [odds ratio (OR) 7.9, 90% confidence interval (90% CI) 1.8-44.9] and worked more often as cleaners (four cases, no referents). Digestive anomalies (OR 11.9, 90% CI 2.0-149) and multiple anomalies (OR 4.5, 90% CI 1.4-16.9) were also associated with occupational exposure to solvents at work. These results were not modified when differences in maternal age, area of residence, and socioeconomic status were taken into account.


Correlation of Bone Mineral Density Scores and Proton Pump Inhibitors Use in the Elderly.
We aimed to identify any correlation between long-term proton pump inhibitor use and low bone mineral density in the elderly. This is a retrospective cohort study looking at PPI usage and bone mineral density scores of patients age 65 and above within a period of two years. Lumbar spine, right hip, and left hip bone mineral density t-scores were reviewed in PPI users and non-users. A total of 1,016 patients who had bone mineral density scans within that period were identified. Non-adjusted data showed no statistically significant association between PPI users and non-users in lumbar spine (p = 0.903), right hip (p = 0.775) and left hip (p = 0.407) bone mineral density scores. After adjusting for age and gender, PPI use was still not associated with low bone mineral density at the lumbar spine (p = 0.957), right hip (p = 0.440), or left hip (p = 0.691). The logistic regression did not show a statistically significant difference in low bone mineral density scores in lumbar spine (p = 0.162), right hip (p = 0.796), and left hip (p = 0.196) when length of use was considered. Among adults 65 years and older proton pump inhibitors users; there appears to be no correlation between PPI use and low bone mineral density and/or the length of PPI use.


A mechanism for initiating RNA-dependent RNA polymerization.
In most RNA viruses, genome replication and transcription are catalysed by a viral RNA-dependent RNA polymerase. Double-stranded RNA viruses perform these operations in a capsid (the polymerase complex), using an enzyme that can read both single- and double-stranded RNA. Structures have been solved for such viral capsids, but they do not resolve the polymerase subunits in any detail. Here we show that the 2 A resolution X-ray structure of the active polymerase subunit from the double-stranded RNA bacteriophage straight phi6 is highly similar to that of the polymerase of hepatitis C virus, providing an evolutionary link between double-stranded RNA viruses and flaviviruses. By crystal soaking and co-crystallization, we determined a number of other structures, including complexes with oligonucleotide and/or nucleoside triphosphates (NTPs), that suggest a mechanism by which the incoming double-stranded RNA is opened up to feed the template through to the active site, while the substrates enter by another route. The template strand initially overshoots, locking into a specificity pocket, and then, in the presence of cognate NTPs, reverses to form the initiation complex; this process engages two NTPs, one of which acts with the carboxy-terminal domain of the protein to prime the reaction. Our results provide a working model for the initiation of replication and transcription.


[Some reflections on body dysmorphic disorder by proxy].
Body dysmorphic disorder by proxy is a body image disorder that causes considerable distress and discomfort to the patient and to his or her close relatives and friends. There have been hardly any publications on this topic and little is know about treatment. In this paper we present the case of a 36-year-old male who exhibited extreme dissatisfaction with his wife's appearance. In particular, cognitive behaviour therapy and several systemic interventions led to a marked reduction in the patient's symptoms.


Protected State Transfer via an Approximate Quantum Adder.
We propose a decoherence protected protocol for sending single photon quantum states through depolarizing channels. This protocol is implemented via an approximate quantum adder engineered through spontaneous parametric down converters, and shows higher success probability than distilled quantum teleportation protocols for distances below a threshold depending on the properties of the channel.


Targeted c-myc gene expression in mammary glands of transgenic mice induces mammary tumours with constitutive milk protein gene transcription.
We investigated the consequences of augmented c-myc gene expression in the mammary gland of transgenic mice. For this purpose we directed the expression of a mouse c-myc transgene to the differentiating mammary epithelial cells by subjecting the protein coding region to the 5' regulatory sequences of the murine whey acidic protein gene (Wap). Analogous to the expression pattern of the endogenous Wap gene, the Wap-myc transgene is abundantly expressed in the mammary gland during lactation. The tissue-specific and hormone-dependent expression of the Wap-myc transgene results in an 80% incidence of mammary adenocarcinomas. As early as two months after the onset of Wap-myc expression, tumours occur in the mammary glands of the transgenic animals. The tumours express not only the Wap-myc transgene, but also the endogenous Wap and beta casein genes. The expression of the milk protein genes becomes independent of the lactogenic hormonal stimuli and persists even in transplanted nude mouse tumours.


MinION rapid sequencing: Review of potential applications in neurosurgery.
Gene sequencing has played an integral role in the advancement and understanding of disease pathology and treatment. Although historically expensive and time consuming, new sequencing technologies improve our capability to obtain the genetic information in an accurate and timely manner. Within neurosurgery, gene sequencing is routinely used in the diagnosis and treatment of neurosurgical diseases, primarily for brain tumors. This paper reviews nanopore sequencing, an innovation utilized by MinION and outlines its potential use for neurosurgery. A literature search was conducted for publications containing the keywords of Oxford MinION, nanopore sequencing, brain tumor, glioma, whole genome sequencing (WGS), epigenomics, molecular neuropathology, and next-generation sequencing (NGS). In total, 64 articles were selected and used for this review. The Oxford MinION nanopore sequencing technology has had successful applications within clinical microbiology, human genome sequencing, and cancer genotyping across multiple specialties. Technical details, methodology, and current use of MinION sequencing are discussed through the prism of potential applications to solve neurosurgery-related scientific and diagnostic questions. The MinION device has proven to provide rapid and accurate reads with longer read lengths when compared with NGS. For applications within neurosurgery, the MinION device is capable of providing critical diagnostic information for central nervous system (CNS) tumors within a single day. MinION provides rapid and accurate gene sequencing with better affordability and convenience compared with current NGS methods. Widespread success of the MinION nanopore sequencing technology in providing accurate, rapid, and convenient gene sequencing suggests a promising future within research laboratories and to improve care for neurosurgical patients.


Triple-bridged ferromagnetic nickel(II) complexes: a combined experimental and theoretical DFT study on stabilization and magnetic coupling.
Two dinuclear [Ni2L2(o-(NO2)C6H4COO)2(H2O)] (1), [Ni2L2(p-(NO2)C6H4COO)2(H2O)]·0.5CH3OH (2) complexes and one trinuclear [Ni3L2(p-(NO2)C6H4COO)4]·C2H5OH (3) Ni(II) complex have been synthesized using a tridentate Schiff base ligand, 1-[(3-dimethylamino-propylimino)-methyl]-naphthalen-2-ol (HL) along with ortho- and para-nitro benzoate as co-ligands. All these three (1-3) complexes have been characterized by spectral analysis, X-ray crystallography and variable temperature magnetic susceptibility measurements. The structural analyses reveal that complexes 1 and 2 are dinuclear in which two μ2-phenoxido and a water molecule bridge the two Ni(ii) centers to make the complexes face sharing bioctahedra. Complex 3 is a linear triple bridged (phenoxido and carboxylato) trinuclear Ni(II) complex. Variable-temperature magnetic susceptibility studies indicate the presence of ferromagnetic exchange coupling in complexes 1-3 with J values of 25.4, 28.1 and 6.2 cm(-1) respectively. Theoretically obtained J values of 21.4 cm(-1) (for 1), 22.0 cm(-1) (for 2) and 9.3 cm(-1) (for 3) corroborate very well the experimental results. DFT calculation also shows that stronger H-bonding interactions present in the case of carboxylate based coligands stabilise the triple oxido-bridged complexes.


Endothelin-1 Mediated Decrease in Mitochondrial Gene Expression and Bioenergetics Contribute to Neurodegeneration of Retinal Ganglion Cells.
Endothelin-1 (ET-1) is a vasoactive peptide that is elevated in aqueous humor as well as circulation of primary open angle glaucoma (POAG) patients. ET-1 has been shown to promote degeneration of optic nerve axons and apoptosis of retinal ganglion cells (RGCs), however, the precise mechanisms are still largely unknown. In this study, RNA-seq analysis was used to assess changes in ET-1 mediated gene expression in primary RGCs, which revealed that 23 out of 156 differentially expressed genes (DEGs) had known or predicted mitochondrial function, of which oxidative phosphorylation emerged as the top-most enriched pathway. ET-1 treatment significantly decreased protein expression of key mitochondrial genes including cytochrome C oxidase copper chaperone (COX17) and ATP Synthase, H+ transporting, Mitochondrial Fo Complex (ATP5H) in primary RGCs and in vivo following intravitreal ET-1 injection in rats. A Seahorse ATP rate assay revealed a significant decrease in the rate of mitochondrial ATP production following ET-1 treatment. IOP elevation in Brown Norway rats showed a trend towards decreased expression of ATP5H. Our results demonstrate that ET-1 produced a decrease in expression of vital components of mitochondrial electron transport chain, which compromise bioenergetics and suggest a mechanism by which ET-1 promotes neurodegeneration of RGCs in glaucoma.


Reversal of vasectomy: the effects of sperm antibodies on subsequent fertility.
Antisperm antibodies were measured in serum and seminal plasma in 130 males before and after vasectomy reversal and the occurrence of pregnancy was analysed in the partners of 77 who were followed for more than one year. Sperm-agglutinating antibodies were found in the serum of 79% of patients; seminal plasma antibodies were present in only 9.5% before reversal and this rose to 26% afterwards. Pregnancies occurred in the partners of 53% of those men who were trying to produce children. A pregnancy was significantly less likely when the pre-operative serum antisperm antibody titre was 512 or more, but no decrease in fertility was seen with titres below this. Several pregnancies were produced by patients with seminal plasma antibodies, but numbers and follow-up are too small to permit detailed analysis. A randomised controlled trial of per-operative steroids showed that they produced no benefit. The antisperm antibodies associated with vasectomy reversal appear to differ fundamentally from those occurring in naturally subfertile males.


Planning the human variome project: the Spain report.
The remarkable progress in characterizing the human genome sequence, exemplified by the Human Genome Project and the HapMap Consortium, has led to the perception that knowledge and the tools (e.g., microarrays) are sufficient for many if not most biomedical research efforts. A large amount of data from diverse studies proves this perception inaccurate at best, and at worst, an impediment for further efforts to characterize the variation in the human genome. Because variation in genotype and environment are the fundamental basis to understand phenotypic variability and heritability at the population level, identifying the range of human genetic variation is crucial to the development of personalized nutrition and medicine. The Human Variome Project (HVP; http://www.humanvariomeproject.org/) was proposed initially to systematically collect mutations that cause human disease and create a cyber infrastructure to link locus specific databases (LSDB). We report here the discussions and recommendations from the 2008 HVP planning meeting held in San Feliu de Guixols, Spain, in May 2008.


Predictive Value of Midsagittal Tissue Bridges on Functional Recovery After Spinal Cord Injury.
The majority of patients with spinal cord injury (SCI) have anatomically incomplete lesions and present with preserved tissue bridges, yet their outcomes vary. To assess the predictive value of the anatomical location (ventral/dorsal) and width of preserved midsagittal tissue bridges for American Spinal Injury Association (ASIA) Impairment Scale (AIS) grade conversion and SCI patient stratification into recovery-specific subgroups. This retrospective longitudinal study includes 70 patients (56 men, age: 52.36 ± 18.58 years) with subacute (ie, 1 month) SCI (45 tetraplegics, 25 paraplegics), 1-month neuroimaging data, and 1-month and 12-month clinical data. One-month midsagittal T2-weighted scans were used to determine the location and width of tissue bridges. Their associations with functional outcomes were assessed using partial correlation and unbiased recursive partitioning conditional inference tree (URP-CTREE). Fifty-seven (81.4%) of 70 patients had tissue bridges (2.53 ± 2.04 mm) at 1-month post-SCI. Larger ventral (P = .001, r = 0.511) and dorsal (P < .001, r = 0.546) tissue bridges were associated with higher AIS conversion rates 12 months post-SCI (n = 39). URP-CTREE analysis identified 1-month ventral tissue bridges as predictors of 12-month total motor scores (0.4 mm cutoff, P = .008), recovery of upper extremity motor scores at 12 months (1.82 mm cutoff, P = .002), 12-month pin-prick scores (1.4 mm cutoff, P = .018), and dorsal tissue bridges at 1 month as predictors of 12-month Spinal Cord Independence Measure scores (0.5 mm cutoff, P = .003). Midsagittal tissue bridges add predictive value to baseline clinical measures for post-SCI recovery. Based on tissue bridges' width, patients can be classified into subgroups of clinical recovery profiles. Midsagittal tissue bridges provide means to optimize patient stratification in clinical trials.


Dividing the mind: the necessary role of the frontal lobes in separating memory from search.
Working memory plays a crucial role in the control of visual selection. Previous research has shown that attentional deployment can be biased to objects in an array matching the contents of working memory. Here, we examined the role of the frontal cortex in determining the interaction between working memory and attention. At the start of each trial, participants memorized an object cue that could contain either the target or a distracter, when the object reappeared in the subsequent search array. Relative to age-matched controls, patients suffering from damage to the frontal lobes showed a stronger effect of the memory stimulus on search. Interestingly, there was an effect of frontal damage on the mean latencies to fixate targets but the effect of memory validity on the number of first saccades to the target, and their time of initiation, was similar across the groups. The results suggest that, following the earliest deployment of attention, frontal lobe structures are involved in separating relevant target from irrelevant (object cue) information, when both are held in memory.


Determinants of Nam8-dependent splicing of meiotic pre-mRNAs.
Nam8, a component of yeast U1 snRNP, is optional for mitotic growth but required during meiosis, because Nam8 collaborates with Mer1 to promote splicing of essential meiotic mRNAs AMA1, MER2 and MER3. Here, we identify SPO22 and PCH2 as novel targets of Nam8-dependent meiotic splicing. Whereas SPO22 splicing is co-dependent on Mer1, PCH2 is not. The SPO22 intron has a non-consensus 5' splice site (5'SS) that dictates its Nam8/Mer1-dependence. SPO22 splicing relies on Mer1 recognition, via its KH domain, of an intronic enhancer 5'-AYACCCUY. Mutagenesis of KH and the enhancer highlights Arg214 and Gln243 and the CCC triplet as essential for Mer1 activity. The Nam8-dependent PCH2 pre-mRNA has a consensus 5'SS and lacks a Mer1 enhancer. For PCH2, a long 5' exon and a non-consensus intron branchpoint dictate Nam8-dependence. Our results implicate Nam8 in two distinct meiotic splicing regulons. Nam8 is composed of three RRM domains, flanked by N-terminal leader and C-terminal tail segments. The leader, tail and RRM1 are dispensable for splicing meiotic targets and unnecessary for vegetative Nam8 function in multiple synthetic lethal genetic backgrounds. Nam8 activity is enfeebled by alanine mutations in the putative RNA binding sites of the RRM2 and RRM3 domains.


Consequences of child emotional abuse, emotional neglect and exposure to intimate partner violence for eating disorders: a systematic critical review.
Child maltreatment and eating disorders are significant public health problems. Yet, to date, research has focused on the role of child physical and sexual abuse in eating-related pathology. This is despite the fact that globally, exposure to emotional abuse, emotional neglect and intimate partner violence are the three of the most common forms of child maltreatment. The objective of the present study is to systematically identify and critically review the literature examining the association between child emotional abuse (EA), emotional neglect (EN), and exposure to intimate partner violence (IPV) and adult eating-disordered behavior and eating disorders. A systematic search was conducted of five electronic databases: Medline, Embase, PsycINFO, CINAHL, and ERIC up to October 2015 to identify original research studies that investigated the association between EA, EN and children's exposure to IPV, with adult eating disorders or eating-disordered behavior using a quantitative research design. Database searches were complemented with forward and backward citation chaining. Studies were critically appraised using the Quality in Prognosis Studies (QUIPS) tool. A total of 5556 publications were screened for this review resulting in twenty-three articles included in the present synthesis. These studies focused predominantly on EA and EN, with a minority examining the role of child exposure to IPV in adult eating-related pathology. Prevalence of EA and EN ranged from 21.0% to 66.0%, respectively. No prevalence information was provided in relation to child exposure to IPV. Samples included predominantly White women. The methodological quality of the available literature is generally low. Currently, the available literature precludes the possibility of determining the extent to which EA, EN or child exposure to IPV have independent explanatory influence in adult eating-related pathology above what has been identified for physical and sexual abuse. While a large proportion of adults with eating disorders or eating-disordered behavior report EA, EN, or child exposure to IPV , there is a paucity of high-quality evidence about these relationships.


Cullin-4 regulates Wingless and JNK signaling-mediated cell death in the Drosophila eye.
In all multicellular organisms, the fundamental processes of cell proliferation and cell death are crucial for growth regulation during organogenesis. Strict regulation of cell death is important to maintain tissue homeostasis by affecting processes like regulation of cell number, and elimination of unwanted/unfit cells. The developing Drosophila eye is a versatile model to study patterning and growth, where complex signaling pathways regulate growth and cell survival. However, the molecular mechanisms underlying regulation of these processes is not fully understood. In a gain-of-function screen, we found that misexpression of cullin-4 (cul-4), an ubiquitin ligase, can rescue reduced eye mutant phenotypes. Previously, cul-4 has been shown to regulate chromatin remodeling, cell cycle and cell division. Genetic characterization of cul-4 in the developing eye revealed that loss-of-function of cul-4 exhibits a reduced eye phenotype. Analysis of twin-spots showed that in comparison with their wild-type counterparts, the cul-4 loss-of-function clones fail to survive. Here we show that cul-4 clones are eliminated by induction of cell death due to activation of caspases. Aberrant activation of signaling pathways is known to trigger cell death in the developing eye. We found that Wingless (Wg) and c-Jun-amino-terminal-(NH2)-Kinase (JNK) signaling are ectopically induced in cul-4 mutant clones, and these signals co-localize with the dying cells. Modulating levels of Wg and JNK signaling by using agonists and antagonists of these pathways demonstrated that activation of Wg and JNK signaling enhances cul-4 mutant phenotype, whereas downregulation of Wg and JNK signaling rescues the cul-4 mutant phenotypes of reduced eye. Here we present evidences to demonstrate that cul-4 is involved in restricting Wg signaling and downregulation of JNK signaling-mediated cell death during early eye development. Overall, our studies provide insights into a novel role of cul-4 in promoting cell survival in the developing Drosophila eye.


Increased pressor responsiveness to enkephalin in spontaneously hypertensive rats: the role of vasopressin.
1. The cardiovascular effects of an enkephalin analogue were examined in spontaneously hypertensive and normotensive Wistar-Kyoto rats. (D-Ala2)-methionine enkephalin caused a biphasic increase in blood pressure and an increase in heart rate after intracerebroventricular injection. 2. The initial pressor response to (D-Ala2)-methionine enkephalin was greater in hypertensive than in normotensive rats. No difference was noted between groups during the secondary pressor response. Heart rate increases paralleled the secondary increase in blood pressure. 3. Naloxone pretreatment abolished the secondary increase in blood pressure and the tachycardia, but did not blunt the initial pressor response in female Wistar-Kyoto rats. 4. Plasma levels of arginine vasopressin were depressed during the plateau phase of the pressor response in hypertensive rats given intracerebroventricular (D-Ala2)-methionine enkephalin. 5. The results suggest that the cardiovascular effects of central enkephalin are not due to vasopressin, but may involve activation of the sympathetic nervous system.


Autism and the Question of the Human.
The article explores how normative notions of emotions and interaction are active in constructions of the categories of "human" and "animal" in different discourses about autism: scientific and autobiographical. In the scientific discourse of autistic emotionality, a deficit perspective of autism is central. The general affective deficit discourse relies on normative discursive notions of "humanity" or "human emotionality." Thus, neurotypicals are produced as real "humans" and neurotypical emotionality as "normal" human emotionality. This human normativity is challenged in the Swedish autobiographical texts by Gunilla Gerland (b. 1963), Iris Johansson (b. 1945) and Immanuel Brändemo (b. 1980). Along with American authors of autobiographies about autism, such as Temple Grandin's Thinking in Pictures (1995) and Dawn Prince-Hughes' Songs of the Gorilla Nation (2004) they destabilize the categories of "human" and "animal" by identifying with nonhuman animals, describing themselves as such, or feeling disqualified as real humans.


Broadband TE-pass slot waveguide polarizer using an asymmetrical directional coupler.
A compact TE-pass polarizer for silicon-based slot waveguides is numerically proposed based on a directional coupler where an asymmetrical slot waveguide (ASW) and hybrid plasmonic waveguide (HPW) are involved. The input section is linearly tapered to an ASW to markedly enlarge the modal birefringence. Beneficially, only the TM mode can be coupled into the adjacent HPW and attenuated while the injected TE mode just passes through the slot waveguide without coupling by properly choosing the dimensions. As a consequence, an efficient TE-pass polarizer can be implemented. From the simulation results, the proposed polarizer has an extinction ratio (ER) of 45 dB and insertion loss (IL) of 0.44 dB at 1.55 μm, and its bandwidth exceeds 130 nm with the ER>30 dB and IL<1 dB. In addition, fabrication tolerances to the key structural parameters are analyzed and field evolution along the propagation distance is also demonstrated.


Add-on bevacizumab can prevent early clinical deterioration and prolong survival in newly diagnosed partially resected glioblastoma patients with a poor performance status.
The AVAglio trial established the beneficial effect of add-on bevacizumab (BEV) for the treatment of newly diagnosed glioblastomas (nd-GBMs) that led to the approval of BEV for the treatment of these patients in Japan. However, the rationality of using BEV as a first-line treatment for nd-GBMs remains controversial. The purpose of this study was to analyze the outcomes of a case series of nd-GBM patients. The outcomes of 69 nd-GBM patients treated after 2006 were retrospectively analyzed. Clinical and genetic analyses were performed, and estimates of progression-free survival (PFS) and overall survival (OS) were calculated using the Kaplan-Meier method. Since add-on BEV therapy was only used for partially resected GBMs (pr-GBMs) after its approval in 2013, the patients were subdivided into 3 treatment groups: Type I, partial removal with temozolomide (TMZ)/BEV and concurrent radiotherapy (CCRT); Type II, partial removal with TMZ and CCRT; and Type III, gross total removal with TMZ and CCRT. The PFS rate of Type I patients was significantly higher than that of Type II patients (P=0.014), but comparable to that of Type III patients. Differences in OS rates between Type I and Type II patients were less apparent (P=0.075), although the median OS of Type I patients was ~8 months higher than that of Type II patients (17.4 vs 9.8 months, respectively). The clinical deterioration rate during initial treatment was significantly (P=0.024) lower in Type I than in Type II patients (7.7% vs 47.4%, respectively). Differences in OS rates between Type I and Type II patients with a poor performance status (PS) were significant (P=0.017). Our findings suggest that add-on BEV can prevent early clinical deterioration of pr-GBM patients and contribute to a prolonged survival, especially for those with a poor PS.


Mapping the Molecular Surface of the Analgesic NaV1.7-Selective Peptide Pn3a Reveals Residues Essential for Membrane and Channel Interactions.
Compelling human genetic studies have identified the voltage-gated sodium channel NaV1.7 as a promising therapeutic target for the treatment of pain. The analgesic spider-venom-derived peptide μ-theraphotoxin-Pn3a is an exceptionally potent and selective inhibitor of NaV1.7; however, little is known about the structure-activity relationships or channel interactions that define this activity. We rationally designed 17 Pn3a analogues and determined their activity at hNaV1.7 using patch-clamp electrophysiology. The positively charged amino acids K22 and K24 were identified as crucial for Pn3a activity, with molecular modeling identifying interactions of these residues with the S3-S4 loop of domain II of hNaV1.7. Removal of hydrophobic residues Y4, Y27, and W30 led to a loss of potency (>250-fold), while replacement of negatively charged D1 and D8 residues with a positively charged lysine led to increased potencies (>13-fold), likely through alterations in membrane lipid interactions. Mutating D8 to an asparagine led to the greatest improvement in Pn3a potency at NaV1.7 (20-fold), while maintaining >100-fold selectivity over the major off-targets NaV1.4, NaV1.5, and NaV1.6. The Pn3a[D8N] mutant retained analgesic activity in vivo, significantly attenuating mechanical allodynia in a clinically relevant mouse model of postsurgical pain at doses 3-fold lower than those with wild-type Pn3a, without causing motor-adverse effects. Results from this study will facilitate future rational design of potent and selective peptidic NaV1.7 inhibitors for the development of more efficacious and safer analgesics as well as to further investigate the involvement of NaV1.7 in pain.


Invited review: piglet mortality: management solutions.
Preweaning mortality varies greatly among herds and this is partly attributed to differences in farrowing house management. In this review, we describe the various management strategies than can be adopted to decrease mortality and critically examine the evidence that exists to support their use. First, we consider which management procedures are effective against specific causes of death: intrapartum stillbirth, hypothermia, starvation, disease, crushing, and savaging. The most effective techniques include intervention to assist dystocic sows, measures to prevent and treat sow hypogalactia, good farrowing house hygiene, providing newborn piglets with a warm microenvironment, early fostering of supernumerary piglets, methods that assist small and weak piglets to breathe and obtain colostrum, and intervention to prevent deaths from crushing and savaging. The provision of nest-building material and modifications to the pen to assist the sow when lying down may also be beneficial, but the evidence is less clear. Because most deaths occur around the time of farrowing and during the first few days of life, the periparturient period is a particularly important time for management interventions intended to reduce piglet mortality. A number of procedures require a stockperson to be present during and immediately after farrowing. Second, we consider the benefits of farrowing supervision for preweaning mortality in general, focusing particularly on methods for the treatment of dystocia and programs of piglet care that combine multiple procedures. Third, we discuss the need for good stockmanship if farrowing supervision is to be effective. Stockmanship refers not only to technical skills but also to the manner in which sows are handled because this influences their fearfulness of humans. We conclude that piglet survival can be improved by a range of management procedures, many of which occur in the perinatal period and require the supervision of farrowing by trained staff. Although this incurs additional labor costs, there is some evidence that this can be economically offset by improved piglet survival.


Requirement of I-E molecule for thymocyte apoptosis induced by staphylococcal enterotoxin B in vivo.
In vivo administration of bacterial superantigen staphylococcal enterotoxin B (SEB) to BALB/c mice led to thymus atrophy resulting from thymocyte apoptosis. In this study, we demonstrated that SEB induced a substantial reduction in thymocyte numbers in BALB/c, B10. D2 (H-2(d) haplotype), B10.BR, C3H/HeJ, C3H/HeN (H-2(k)), and (BALB/c x B6)F1 (H-2(dxb)), but caused little or no effect in I-E- strains such as B6, B10, A.BY (H-2(b)), and A.SW (H-2(s)) mice. Elimination of CD4(+)CD8(+) cells predominantly accounted for the thymocyte loss, although the numbers of other subpopulations may also be reduced. Thymocyte apoptosis was shown by an increase in the level of DNA fragmentation in BALB/c but not in B6 mice after SEB administration. Treatment with anti-I-Ed monoclonal antibody to BALB/c mice blocked SEB-induced thymocyte apoptosis when anti-I-Ad exerted less effect. In contrast to SEB, staphylococcal enterotoxin A led to comparable levels of thymus atrophy in BALB/c and B6 mice. Studies on the surface marker expression indicated that CD25 expression was upregulated on BALB/c mouse thymocytes but with only a moderate increase in B6 mice. The CD4(+)CD8(+) cells were the major (>90%) population that expressed elevated levels of CD25 in BALB/c mice. An increase in the expression of TCRalphabeta, CD3, and CD69 surface markers was also observed on thymocytes from BALB/c mice, but not from I-E- strains. The differential response of I-E+ and I-E- mice to SEB may be exploited as a model for the study of apoptosis in the thymus.


No association of multiple sclerosis activity and progression with EBV or tobacco use in BENEFIT.
To evaluate whether Epstein-Barr virus (EBV) immunoglobulin G (IgG) antibody levels or tobacco use were associated with conversion to multiple sclerosis (MS) or MS progression/activity in patients presenting with clinically isolated syndrome (CIS). In this prospective, longitudinal study, we measured EBV IgG antibody and cotinine (biomarker of tobacco use) levels at up to 4 time points (baseline, months 6, 12, and 24) among 468 participants with CIS enrolled in the BENEFIT (Betaferon/Betaseron in Newly Emerging Multiple Sclerosis for Initial Treatment) clinical trial. Outcomes included time to conversion to clinically definite or McDonald MS, number of relapses, Expanded Disability Status Scale (EDSS) changes, brain and T2 lesion volume changes, and number of new active lesions over 5 years. Analyses were adjusted for age, sex, treatment allocation, baseline serum 25-hydroxyvitamin D level, number of T2 lesions, body mass index, EDSS, steroid treatment, and CIS onset type. We found no associations between any EBV IgG antibody or cotinine levels with conversion from CIS to MS or MS progression as measured by EDSS or activity clinically or on MRI. The relative risk of conversion from CIS to clinically definite MS was 1.14 (95% confidence interval 0.76-1.72) for the highest vs the lowest quintile of EBNA-1 IgG levels, and 0.96 (95% confidence interval 0.71-1.31) for cotinine levels >25 ng/mL vs <10. Neither increased levels of EBV IgG antibodies, including against EBNA-1, nor elevated cotinine levels indicative of tobacco use, were associated with an increased risk of CIS conversion to MS, or MS activity or progression over a 5-year follow-up.


Neolithic mitochondrial haplogroup H genomes and the genetic origins of Europeans.
Haplogroup H dominates present-day Western European mitochondrial DNA variability (>40%), yet was less common (~19%) among Early Neolithic farmers (~5450 BC) and virtually absent in Mesolithic hunter-gatherers. Here we investigate this major component of the maternal population history of modern Europeans and sequence 39 complete haplogroup H mitochondrial genomes from ancient human remains. We then compare this 'real-time' genetic data with cultural changes taking place between the Early Neolithic (~5450 BC) and Bronze Age (~2200 BC) in Central Europe. Our results reveal that the current diversity and distribution of haplogroup H were largely established by the Mid Neolithic (~4000 BC), but with substantial genetic contributions from subsequent pan-European cultures such as the Bell Beakers expanding out of Iberia in the Late Neolithic (~2800 BC). Dated haplogroup H genomes allow us to reconstruct the recent evolutionary history of haplogroup H and reveal a mutation rate 45% higher than current estimates for human mitochondria.


Percutaneous angioplasty and stenting of the superficial femoral artery.
The objectives of this study were to examine factors predictive of success or failure after percutaneous angioplasty (PTA) and stenting (S) of the superficial femoral artery (SFA) and to compare the results of PTA/S with a contemporary group of patients treated with femoropopliteal bypass. A database of patients undergoing PTA and/or S of the SFA between 1986 and 2004 was maintained. Intention-to-treat analysis was performed. Patients underwent duplex scanning follow-up at 1, 3, and every 6 months after the intervention. Angiograms were reviewed in all cases to assess lesion characteristics and preprocedure and postprocedure runoff. Results were standardized to current TransAtlantic Inter-Society Consensus (TASC) and Society for Vascular Surgery (SVS) criteria. Kaplan-Meier survival analyses were performed to assess time-dependent outcomes. Cox proportional hazard analyses were performed to assess factors associated with patient survival and treatment efficacy. Three hundred eighty total limbs underwent PTA/S in 329 patients (67% male, 33% female; average age, 65 years). Mean follow-up was 1.8 years from the date of initial intervention. Indications for intervention were claudication in 66%, rest pain in 16%, and tissue loss in 18%. Runoff at the tibial level was 2.1 +/- 0.8 patent vessels. Mean SVS ischemia grade was 3.1 (range, 1 to 5). TASC lesion grades were A (48%), B (18%), C (22%), and D (12%). Angioplasty alone was used in 63% of cases. Primary treatment failure (inability to cross lesion) was seen in 7% of patients. There was one periprocedural death. Primary patency rates were 86% at 3 months, 80% at 6 months, 75% at 12 months, 66% at 24 months, 60% at 36 months, 58% at 48 months, and 52% at 60 months. Assisted primary patency rates were slightly higher ( P = not significant). By Cox proportional hazards analysis, patency of PTA/S was associated with higher preoperative ankle/brachial index ( P = .016) and the performance of angioplasty only ( P = .011). Failed or occluded PTA/S was associated with TASC C ( P < .0001) and TASC D lesions ( P < .0001). Patient death was associated with the presence of congestive heart failure ( P = .003). Subgroup analysis revealed that primary patency rates are highly dependent on lesion type (A > B > C > D, P < .0001). PTA/S patency for TASC A and B lesions compared favorably to prosthetic and venous femoropopliteal bypass. Surgical bypass was superior to PTA/S for TASC C and D lesions. PTA and stenting of the SFA can be performed safely with excellent procedural success rates. Improved patency of these interventions was seen with increased ankle/brachial index and the performance of angioplasty only. Worse patency was seen with TASC C and TASC D lesions. Patency rates were strongly dependent on lesion type, and the results of angioplasty and stenting compared favorably with surgical bypass for TASC A and B lesions.


14-3-3 amplifies and prolongs adrenergic stimulation of HERG K+ channel activity.
Acute stress provokes lethal cardiac arrhythmias in the hereditary long QT syndrome. Here we provide a novel molecular mechanism linking beta-adrenergic signaling and altered human ether-a-go-go related gene (HERG) channel activity. Stress stimulates beta-adrenergic receptors, leading to cAMP elevations that can regulate HERG K+ channels both directly and via phosphorylation by cAMP-dependent protein kinase (PKA). We show that HERG associates with 14-3-3epsilon to potentiate cAMP/PKA effects upon HERG. The binding of 14-3-3 occurs simultaneously at the N- and C-termini of the HERG channel. 14-3-3 accelerates and enhances HERG activation, an effect that requires PKA phosphorylation of HERG and dimerization of 14-3-3. The interaction also stabilizes the lifetime of the PKA-phosphorylated state of the channel by shielding the phosphates from cellular phosphatases. The net result is a prolongation of the effect of adrenergic stimulation upon HERG activity. Thus, 14-3-3 interactions with HERG may provide a unique mechanism for plasticity in the control of membrane excitability and cardiac rhythm.


Double-digest RAD sequencing outperforms microsatellite loci at assigning paternity and estimating relatedness: A proof of concept in a highly promiscuous bird.
Information on genetic relationships among individuals is essential to many studies of the behaviour and ecology of wild organisms. Parentage and relatedness assays based on large numbers of single nucleotide polymorphism (SNP) loci hold substantial advantages over the microsatellite markers traditionally used for these purposes. We present a double-digest restriction site-associated DNA sequencing (ddRAD-seq) analysis pipeline that, as such, simultaneously achieves the SNP discovery and genotyping steps and which is optimized to return a statistically powerful set of SNP markers (typically 150-600 after stringent filtering) from large numbers of individuals (up to 240 per run). We explore the trade-offs inherent in this approach through a set of experiments in a species with a complex social system, the variegated fairy-wren (Malurus lamberti) and further validate it in a phylogenetically broad set of other bird species. Through direct comparisons with a parallel data set from a robust panel of highly variable microsatellite markers, we show that this ddRAD-seq approach results in substantially improved power to discriminate among potential relatives and considerably more precise estimates of relatedness coefficients. The pipeline is designed to be universally applicable to all bird species (and with minor modifications to many other taxa), to be cost- and time-efficient, and to be replicable across independent runs such that genotype data from different study periods can be combined and analysed as field samples are accumulated.


Comprehensive predictions of target proteins based on protein-chemical interaction using virtual screening and experimental verifications.
Identification of the target proteins of bioactive compounds is critical for elucidating the mode of action; however, target identification has been difficult in general, mostly due to the low sensitivity of detection using affinity chromatography followed by CBB staining and MS/MS analysis. We applied our protocol of predicting target proteins combining in silico screening and experimental verification for incednine, which inhibits the anti-apoptotic function of Bcl-xL by an unknown mechanism. One hundred eighty-two target protein candidates were computationally predicted to bind to incednine by the statistical prediction method, and the predictions were verified by in vitro binding of incednine to seven proteins, whose expression can be confirmed in our cell system.As a result, 40% accuracy of the computational predictions was achieved successfully, and we newly found 3 incednine-binding proteins. This study revealed that our proposed protocol of predicting target protein combining in silico screening and experimental verification is useful, and provides new insight into a strategy for identifying target proteins of small molecules.


Reduction of Pain and Spinal Nociceptive Transmission by Working Memory is Load Dependant.
Working memory (WM) engagement produces pain inhibition. However, it remains unclear whether higher WM load increases this effect. The aim of this study was to investigate the interaction between WM load and pain inhibition by WM and examine the contribution of cerebrospinal mechanism. Thirty-eight healthy volunteers were assigned to one of 2 n-back groups for which WM load was different (2-back or 3-back). The experimental protocol comprised 5 counterbalanced conditions (0-back, n-back, pain, 0-back with pain, and n-back with pain). Pain and the nociceptive flexion reflex (NFR) were evoked by transcutaneous electrical stimulation of the sural nerve. Pain was significantly different between conditions, but not between n-back groups. Both the 0-back and n-back tasks reduced pain compared with pain alone, but the n-back task produced stronger pain inhibition compared with the 0-back task. NFR amplitude was significantly different between conditions but not between n-back groups. NFR was inhibited by the 0-back and n-back tasks, with no difference between the 2 tasks. These findings indicate that pain inhibition by WM is increased by WM load, but only to a certain point. NFR inhibition by WM suggests that inhibition of pain by WM depends, at least in part, on cerebrospinal mechanism. PERSPECTIVE: This behavioral and electrophysiological study shows that engaging in a cognitive task reduces pain by decreasing spinal nociceptive transmission, depending on task difficulty. These findings may yield better nonpharmacological pain therapies based on individual differences in working memory performance and capacity as well as several factors that regulate working memory.


"Like an uncle but more, but less than a father"--Irish children's relationships with nonresident fathers.
Although previous research has highlighted the importance of the quality of nonresident father-child relationships for children's well-being, little is known about children's perspectives on what underpins feelings of closeness to their nonresident fathers. This qualitative study explored the processes that facilitate or constrain children's feelings of closeness to their nonresident fathers. Semistructured interviews were conducted with 27 children (ages 8 to 17) who had grown up in a single-mother household, where fathers were nonresident from early in the child's life. Findings revealed the fragility of children's ties with their nonresident fathers and the risk that nonresidence from the outset placed upon these relationships. Children's experiences of closeness to fathers were related to perceptions of their fathers' commitment to their relationship and his obligation to his parenting role, and to a sense of connection to and familiarity with their fathers. It was a challenge for children to feel connected to their fathers when contact arrangements were detached from caregiving activities and precluded immersion in each other's daily lives. Lack of effort on the part of fathers to maintain contact or failure to keep arrangements constrained children's feelings of closeness and gave rise to feelings of disappointment and anger. Children demonstrated their capacity to act as agents within their families as they made sense of these relationships for themselves and accepted or rejected their father as a person who could play a meaningful role in their lives. The implications of the findings for promoting positive relationships with nonresident fathers are discussed.


Factors associated with non-adherence to three hypertension self-management behaviors: preliminary data for a new instrument.
Clinicians have difficulty in identifying patients that are unlikely to adhere to hypertension self-management. Identifying non-adherence is essential to addressing suboptimal blood pressure control and high costs. 1) To identify risk factors associated with non-adherence to three key self-management behaviors in patients with hypertension: proper medication use, diet, and exercise; 2) To evaluate the extent to which an instrument designed to identify the number of risk factors present for non-adherence to each of the three hypertension self-management behaviors would be associated with self-management non-adherence and blood pressure. Cross-sectional analysis of randomized trial data. Six hundred and thirty-six primary care patients with hypertension. 1) Demographic, socioeconomic, psychosocial, and health belief-related factors; 2) measures of self-reported adherence to recommended medication use, diet recommendations, and exercise recommendations, all collected at baseline assessment; 3) systolic blood pressure (SBP) and diastolic blood pressure (DBP). We identified patient factors associated with measures of non-adherence to medications, diet, and exercise in hypertension. We then combined risk factors associated with ≥1 adherence measure into an instrument that generated three composite variables (medication, diet, and exercise composites), reflecting the number of risk factors present for non-adherence to the corresponding self-management behavior. These composite variables identified subgroups with higher likelihood of medication non-adherence, difficulty following diet recommendations, and difficulty following exercise recommendations. Composite variable levels representing the highest number of self-management non-adherence risk factors were associated with higher SBP and DBP. We identified factors associated with measures of non-adherence to recommended medication use, diet, and exercise in hypertension. We then developed an instrument that was associated with non-adherence to these self-management behaviors, as well as with blood pressure. With further study, this instrument has potential to improve identification of non-adherent patients with hypertension.


Do positional variations of epididymis affect infertility?
To determine the ratio of epididymal position variations and whether this had any effect on seminal parameters. Scrotal examination results and epididymal positions were retrospectively assessed in subjects presenting to our clinic for infertility, varicocele, and sexual developmental delay. Scrotal examination consisted of epididymal position, testicular volumes, and vas deferens and varicocele presence. The subjects were categorized according to sperm counts per mL. Data were assessed whether there was a significant difference in terms of epididymal positions among the groups. A total of 5812 epididymides were examined. There was no significant difference with respect to one or two epididymides being in an anterior position between the groups allocated with respect to a sperm count above 15 million per mL, between 5 and 15 million per mL, and below 5 million per mL (p=0.542). Sperm counts and motility did not differ according to the position of the epididymis in analyzes performed separately within each group. Positional variations of epididymis can be seen at a considerable rate. Nevertheless, there was no clear relationship between these variations and infertility. In terms of avoiding possible misdiagnoses and treatment complications possible epididymal position variations should be kept in mind during clinical practice.


Ischemia induces regulator of G protein signaling 2 (RGS2) protein upregulation and enhances apoptosis in astrocytes.
Regulator of G protein signaling (RGS) family members, such as RGS2, interact with Galpha subunits of heterotrimeric G proteins, accelerating the rate of GTP hydrolysis and attenuating the intracellular signaling triggered by the G protein-coupled receptor-ligand interaction. They are also reported to regulate G protein-effector interactions and form multiprotein signaling complexes. Ischemic stress-induced changes in RGS2 expression have been described in astrocytes, and these changes are associated with intracellular signaling cascades, suggesting that RGS2 upregulation may be an important mechanism by which astrocytes may regulate RGS2 function in response to physiological stress. However, information on the functional roles of stress-induced modulation of RGS2 protein expression in astrocyte function is limited. We report the role of ischemic stress in RGS2 protein expression in rat C6 astrocytoma cells and primary mouse astrocytes. A marked increase in RGS2 occurred after ischemic stress induced by chemicals (sodium azide and 2-deoxyglucose) or oxygen-glucose deprivation (OGD, real ischemia). RGS2 mRNA expression was markedly enhanced by 1 h of exposure to chemical ischemia or 6 h of OGD followed by 2 or 6 h of recovery, respectively. This enhanced expression in primary astrocytes and C6 cells was restored to baseline levels after 12 h of recovery from chemically induced ischemic stress or 4-6 h of recovery from OGD. RGS2 protein was also significantly expressed at 12-24 h of recovery from ischemic insult. Ischemia-induced RGS2 upregulation was associated with enhanced apoptosis. It significantly increased annexin V-positive cells, cleaved caspase-3, and enhanced DNA ladder formation and cell cycle arrest. However, a small interfering RNA (siRNA)-mediated RGS2 knockdown reversed the apoptotic cell death associated with ischemia-induced RGS2 upregulation. Upregulated RGS2 was significantly inhibited by SB-203580, a p38 MAPK inhibitor. Rottlerin, a potent inhibitor of PKCdelta, completely abrogated the increased RGS2 expression. We also examine whether ischemia-induced RGS2-mediated apoptosis is affected by siRNA-targeted endogenous PKCdelta downregulation or its phosphorylation. Although RGS2 upregulation was not affected, siRNA transfection significantly suppressed endogenous PKCdelta mRNA and protein expressions. Ischemia-induced PKCdelta phosphorylation and caspase-3 cleavage were dose dependently inhibited by PKCdelta knockdown, and this endogenous PKCdelta suppression reversed ischemia-induced annexin V-positive cells. This study suggests that ischemic stress increases RGS2 expression and that this condition contributes to enhanced apoptosis in C6 cells and primary astrocytes. The signaling it follows may involve PKCdelta and p38 MAPK pathways.


Sugar feeding by invasive mosquito species on ornamental and wild plants.
Feeding on plant-derived sugars is an essential component of mosquito biology, affecting key aspects of their lives such as survival, metabolism, and reproduction. Among mosquitoes, Aedes aegypti and Aedes albopictus are two invasive mosquito species in the US, and are vectors of diseases such as dengue fever, chikungunya, and Zika. These species live in heavily populated, urban areas, where they have high accessibility to human hosts as well as to plants in backyards and public landscapes. However, the range of plants that are suitable sugar hosts for these species remains to be described, despite the importance of understanding what plants may attract or repel mosquitoes to inform citizens and municipal authorities accordingly. Here, we tested whether Ae. aegypti and Ae. albopictus would sugar-feed on eleven commonly planted ornamental plant species. We confirmed feeding activity using the anthrone method and identified the volatile composition of plant headspace using gas-chromatography mass-spectroscopy. These chemical analyses revealed that a broad range of olfactory cues are associated with plants that mosquitoes feed on. This prompted us to use plant DNA barcoding to identify plants that field-caught mosquitoes feed on. Altogether, results show that native and invasive mosquito species can exploit a broader range of plants than originally suspected, including wild and ornamental plants from different phyla throughout the Spring, Summer and Fall seasons.


Angioarchitecture of arteriovenous fistulas at the craniocervical junction: a multicenter cohort study of 54 patients.
OBJECTIVE The aim of this retrospective multicenter cohort study was to assess the details of the angioarchitecture of arteriovenous fistulas (AVFs) at the craniocervical junction (CCJ) and to determine the associations between the angiographic characteristics and the clinical presentations and outcomes. METHODS The authors analyzed angiographic and clinical data for patients with CCJ AVFs from 20 participating centers that are members of the Japanese Society for Neuroendovascular Therapy (JSNET). Angiographic findings (feeding artery, location of AV shunt, draining vein) and patient data (age, sex, presentation, treatment modality, outcome) were tabulated and stratified based on the angiographic types of the lesions, as diagnosed by a member of the CCJ AVF study group, which consisted of a panel of 6 neurointerventionalists and 1 spine neurosurgeon. RESULTS The study included 54 patients (median age 65 years, interquartile range 61-75 years) with a total of 59 lesions. Five angiographic types were found among the 59 lesions: Type 1, dural AVF (22 [37%] of 59); Type 2, radicular AVF (17 [29%] of 59); Type 3, epidural AVF (EDAVF) with pial feeders (8 [14%] of 59); Type 4, EDAVF (6 [10%] of 59); and Type 5, perimedullary AVF (6 [10%] of 59). In almost all lesions (98%), AV shunts were fed by radiculomeningeal arteries from the vertebral artery that drained into intradural or epidural veins through AV shunts on the dura mater, on the spinal nerves, in the epidural space, or on the spinal cord. In more than half of the lesions (63%), the AV shunts were also fed by a spinal pial artery from the anterior spinal artery (ASA) and/or the lateral spinal artery. The data also showed that the angiographic characteristics associated with hemorrhagic presentations-the most common presentation of the lesions (73%)-were the inclusion of the ASA as a feeder, the presence of aneurysmal dilatation on the feeder, and CCJ AVF Type 2 (radicular AVF). Treatment outcomes differed among the angiographic types of the lesions. CONCLUSIONS Craniocervical junction AVFs commonly present with hemorrhage and are frequently fed by both radiculomeningeal and spinal pial arteries. The AV shunt develops along the C-1 or C-2 nerve roots and can be located on the spinal cord, on the spinal nerves, and/or on the inner or outer surface of the dura mater.


Accurate and fast urinalysis in febrile patients by flow cytometry.
The urine culture is worldwide accepted as the gold standard in diagnosing urinary tract infections, but is time consuming and costly, other methods are fast but moderately reliable. We investigated whether counting the number of bacteria by flow cytometry could be a fast and accurate method to analyze urine samples in febrile patients at the emergency department (ED). Urine samples were obtained from 140 febrile patients at the ED. Urinalysis was performed according to standard procedures. Flow cytometric analysis for bacteria was performed with the Accuri C6 flow cytometer. Diagnostic values were determined at various cut-off points by using urine culture as the gold standard. The highest diagnostic accuracy of urinalysis of bacteria was obtained with flow cytometric analysis (AUC of 0.96). The best cut-off value for bacteria counted by flow cytometry based on the ROC-curve was 3.72 × 106 bacteria/mL, this resulted in a sensitivity of 94.7% and a specificity of 88.2%. Counting bacteria by flow cytometry has the highest diagnostic accuracy and is superior to other methods in urinalysis in febrile patients in the ED when using urine culture as the gold standard.


Apathy, but not depression, reflects inefficient cognitive strategies in Parkinson's disease.
The relationship between apathy, depression and cognitive impairment in Parkinson's disease (PD) is still controversial. The objective of this study is to investigate whether apathy and depression are associated with inefficient cognitive strategies in PD. In this prospective clinical cohort study conducted in a university-based clinical and research movement disorders center we studied 48 PD patients. Based on clinical evaluation, they were classified in two groups: PD with apathy (PD-A group, n = 23) and PD without apathy (PD-NA group, n = 25). Patients received clinical and neuropsychological evaluations. The clinical evaluation included: Apathy Evaluation Scale-patient version, Hamilton Depression Rating Scale-17 items, the Unified Parkinson's Disease Rating Scale and the Hoehn and Yahr staging system; the neuropsychological evaluation explored speed information processing, attention, working memory, executive function, learning abilities and memory, which included several measures of recall (immediate free, short delay free, long delay free and cued, and total recall). PD-A and PD-NA groups did not differ in age, disease duration, treatment, and motor condition, but differed in recall (p<0.001) and executive tasks (p<0.001). Immediate free recall had the highest predictive value for apathy (F = 10.94; p = 0.002). Depression and apathy had a weak correlation (Pearson index= 0.3; p<0.07), with three items of the depression scale correlating with apathy (Pearson index between .3 and.4; p<0.04). The depressed and non-depressed PD patients within the non-apathetic group did not differ. Apathy, but not depression, is associated with deficit in implementing efficient cognitive strategies. As the implementation of efficient strategies relies on the fronto-striatal circuit, we conclude that apathy, unlike depression, is an early expression of executive impairment in PD.