Renal blood flow and oxygenation drive nephron progenitor differentiation.
During kidney development, the vasculature develops via both angiogenesis (branching from major vessels) and vasculogenesis (de novo vessel formation). The formation and perfusion of renal blood vessels are vastly understudied. In the present study, we investigated the regulatory role of renal blood flow and O2 concentration on nephron progenitor differentiation during ontogeny. To elucidate the presence of blood flow, ultrasound-guided intracardiac microinjection was performed, and FITC-tagged tomato lectin was perfused through the embryo. Kidneys were costained for the vasculature, ureteric epithelium, nephron progenitors, and nephron structures. We also analyzed nephron differentiation in normoxia compared with hypoxia. At embryonic day 13.5 (E13.5), the major vascular branches were perfused; however, smaller-caliber peripheral vessels remained unperfused. By E15.5, peripheral vessels started to be perfused as well as glomeruli. While the interior kidney vessels were perfused, the peripheral vessels (nephrogenic zone) remained unperfused. Directly adjacent and internal to the nephrogenic zone, we found differentiated nephron structures surrounded and infiltrated by perfused vessels. Furthermore, we determined that at low O2 concentration, little nephron progenitor differentiation was observed; at higher O2 concentrations, more differentiation of the nephron progenitors was induced. The formation of the developing renal vessels occurs before the onset of blood flow. Furthermore, renal blood flow and oxygenation are critical for nephron progenitor differentiation.


The use of cytokeratin 7 and EMA in differentiating ovarian yolk sac tumors from endometrioid and clear cell carcinomas.
Yolk sac tumors (YSTs) have a variety of morphologic patterns, some of which can resemble either endometrioid adenocarcinoma (EAC) or clear cell carcinoma (CCC). Immunohistochemical staining for alpha-fetoprotein (AFP) is usually only focal and thus is not always helpful in the diagnosis of YST, and pancytokeratin (CK) is expressed by all three tumors. We studied a battery of immunohistochemical markers with specific attention to the utility of cytokeratin 7 (CK7) in differentiating YST from EAC and CCC. A total of 46 ovarian tumors were retrieved for this study: 16 YST, 19 EAC, and 11 CCC. The three groups were analyzed for the expression of CK7, AFP, Leu-M1 (CD15), EMA, and WT1 by immunohistochemistry. In addition, CK and c-kit (CD117) were studied in the YSTs. All of the YSTs tested (100%) were positive for CK. CK7 was considered negative in all 16 YST cases (100%), although a few tumor cells (1%-2%) stained in 4 cases. In contrast, 17 of 19 EACs and all 11 CCCs had diffuse 3+ to 4+ positivity for CK7; the two other EACs showed 2+ positivity for CK7 (40% and 30% of the tumors). AFP was positive in 12 of 15 YSTs (80%), but was generally focal with 1+ staining in 10 cases (67%); only 2 cases were 3+. All of the EACs and CCCs were negative for AFP. Leu-M1 was 1+ in 9 of 15 YSTs (60%), while the remaining 6 were considered negative. Leu-M1 was positive in 10 of 15 EACs tested (67%), but the staining was variable with 1 case 3+, 3 cases 2+, and 6 cases 1+. In the CCCs, 10 cases (91%) were 3+ to 4+, and 1 case was 1+. EMA was essentially negative in 15 of 15 YSTs (100%), with 3 completely negative and 12 showing very focal (<5%) staining. Eight of 12 EACs showed 4+ staining, 3 showed 3+ staining, and 1 showed 2+ staining. All of the 11 CCCs (100%) showed 4+ staining. WT1 was negative in all cases of YST and CCC; 16 of 18 EAC tested (89%) were negative for WT1, but 2 (11%) were 4+ positive. C-kit was negative in all YSTs. In conclusion, it is important for pathologists to be aware that YSTs may mimic EACs and CCCs and that this distinction is important for the clinical management of patients with these tumors. AFP staining is focal in most YST, so an absence of staining does not exclude this diagnosis. CK7 and EMA are essentially negative in YST but are diffusely positive in CCC and EAC, making them useful markers for differentiating YSTs from both CCCs and EACs. Leu-M1 may also be helpful for distinguishing YSTs from CCCs.


The effects of positive airway pressure on cardiovascular responses, perceived effort and time to exhaustion during high-intensity exercise in healthy subjects.
We investigated the effects of continuous positive airway pressure (CPAP) on the time to fatigue, rates of perceived exertion (RPE), and cardiovascular responses during sustained, high-intensity exercise (80% of VO2peak). Healthy male subjects (N.=11) underwent CPAP adaptation at rest to exclude those who felt incapable of breathing with the device and to determine the hemodynamic-induced changes. Subjects performed two exercise sessions, pedaling at 80% of VO2peak with or without CPAP until volitional fatigue. Heart rate was measured continuously, and blood pressure and RPE were recorded every minute. The time to exhaustion was increased in the sessions with CPAP compared with sessions without CPAP (889±270 s vs. 689±256 sec, respectively, P=0.037). There were no differences in the cardiovascular variables at the end of the exercise sessions with or without CPAP. The RPE was lower during exercise with CPAP, but it reached maximal values similar to those observed at the end of exercise without CPAP. During sustained, high-intensity exercise (80% of VO2peak), CPAP extended the time to fatigue in healthy subjects but did not change the heart rate, blood pressure or rate pressure product. Additionally, the central and peripheral RPEs remained lower for a longer period of time during exercise with CPAP.


PCMD-1 bridges the centrioles and the pericentriolar material scaffold in C. elegans.
Correct cell division relies on the formation of a bipolar spindle. In animal cells, microtubule nucleation at the spindle poles is facilitated by the pericentriolar material (PCM), which assembles around a pair of centrioles. Although centrioles are essential for PCM assembly, the proteins that anchor the PCM to the centrioles are less known. Here, we investigate the molecular function of PCMD-1 in bridging the PCM and the centrioles in Caenorhabditis elegans. We demonstrate that the centrosomal recruitment of PCMD-1 is dependent on the outer centriolar protein SAS-7. The most C-terminal part of PCMD-1 is sufficient to target it to the centrosome, and the coiled-coil domain promotes its accumulation by facilitating self-interaction. We reveal that PCMD-1 interacts with the PCM scaffold protein SPD-5, the mitotic kinase PLK-1 and the centriolar protein SAS-4. Using an ectopic translocation assay, we show that PCMD-1 can selectively recruit downstream PCM scaffold components to an ectopic location in the cell, indicating that PCMD-1 is able to anchor the PCM scaffold proteins at the centrioles. Our work suggests that PCMD-1 is an essential functional bridge between the centrioles and the PCM.


Untangling the evolutionary history of a highly polymorphic species: introgressive hybridization and high genetic structure in the desert cichlid fish Herichtys minckleyi.
Understanding the origin of biodiversity requires knowledge on the evolutionary processes that drive divergence and speciation, as well as on the processes constraining it. Intraspecific polymorphisms can provide insight into the mechanisms that generate and maintain phenotypic, behavioural and life history diversification, and can help us understand not only the processes that lead to speciation but also the processes that prevent local fixation of morphs. The 'desert cichlid' Herichtys minckleyi is a highly polymorphic species endemic to a biodiversity hotspot in northern Mexico, the Cuatro Ciénegas valley. This species is polymorphic in body shape and trophic apparatus, and eco-morphotypes coexist in small spring-fed lagoons across the valley. We investigated the genetic structure of these polymorphisms and their phylogeographic history by analysing the entire control region of the mitochondrial DNA and 10 nuclear microsatellite markers in several populations from different sites and morphs. We found two very divergent mitochondrial lineages that most likely predate the closing of the valley and are not associated with morphotypes or sites. One of these lineages is also found in the sister species Herichthys cyanoguttatus. Data from neutral microsatellite markers suggest that most lagoons or drainages constitute their own genetic cluster with sympatric eco-morphotypes forming panmictic populations. Alternative mechanisms such as phenotypic plasticity and a few loci controlled traits provide possible explanations for the sympatric coexistence of discrete nonoverlapping eco-morphotypes with apparent lack of barriers to gene flow within multiple lagoons and drainages.


Unconscious and Distinctive Control of Vocal Pitch and Timbre During Altered Auditory Feedback.
Vocal control plays a critical role in smooth social communication. Speakers constantly monitor auditory feedback (AF) and make adjustments when their voices deviate from their intentions. Previous studies have shown that when certain acoustic features of the AF are artificially altered, speakers compensate for this alteration in the opposite direction. However, little is known about how the vocal control system implements compensations for alterations of different acoustic features, and associates them with subjective consciousness. The present study investigated whether compensations for the fundamental frequency (F0), which corresponds to perceived pitch, and formants, which contribute to perceived timbre, can be performed unconsciously and independently. Forty native Japanese speakers received two types of altered AF during vowel production that involved shifts of either only the formant frequencies (formant modification; Fm) or both the pitch and formant frequencies (pitch + formant modification; PFm). For each type, three levels of shift (slight, medium, and severe) in both directions (increase or decrease) were used. After the experiment, participants were tested for whether they had perceived a change in the F0 and/or formants. The results showed that (i) only formants were compensated for in the Fm condition, while both the F0 and formants were compensated for in the PFm condition; (ii) the F0 compensation exhibited greater precision than the formant compensation in PFm; and (iii) compensation occurred even when participants misperceived or could not explicitly perceive the alteration in AF. These findings indicate that non-experts can compensate for both formant and F0 modifications in the AF during vocal production, even when the modifications are not explicitly or correctly perceived, which provides further evidence for a dissociation between conscious perception and action in vocal control. We propose that such unconscious control of voice production may enhance rapid adaptation to changing speech environments and facilitate mutual communication.


Skeletal morphogenesis in the sea urchin embryo: regulation of primary mesenchyme gene expression and skeletal rod growth by ectoderm-derived cues.
The skeleton of the sea urchin embryo is synthesized by the primary mesenchyme cells (PMCs). Previous studies have shown that local interactions between PMCs and the neighboring ectoderm regulate several aspects of skeletal morphogenesis, including PMC distribution in the blastocoel, the size of the skeleton and its branching pattern. In the present study, we have further examined the regulation of skeletogenesis by the ectoderm. We generated a 'rate map' of skeletal growth, which revealed stereotypical changes in the rates at which specific skeletal elements elongate during development. We showed that three transcripts encoding PMC-specific gene products known to be involved in the synthesis of the skeleton exhibited dynamic, spatially regulated patterns of expression within the PMC syncytium. All three gene products showed high levels of expression at sites of skeletal rod growth, although the specific patterns varied among the genes. We present direct evidence, based upon cell transplantation experiments, that the expression of one of these genes, SM30, is responsive to local, ectoderm-derived cues. Based upon our studies, we suggest that short-range signals from different ectodermal territories may regulate the expression of PMC-specific gene products that are rate-limiting in skeletal biosynthesis, thereby locally influencing skeletal rod growth.


Systolic blood pressure and mortality in prevalent haemodialysis patients in the HEMO study.
Previous studies of blood pressure and mortality in haemodialysis have yielded mixed results, perhaps due to confounding by comorbid conditions. We hypothesized that after improved accounting for confounding factors, higher systolic blood pressure (SBP) would be associated with higher all-cause mortality. We conducted a secondary analysis of data from the haemodialysis study, a randomized trial in prevalent haemodialysis patients. We used three proportional hazard models to determine the relative hazard at different levels of SBP: (1) Model-BL used baseline SBP; (2) Model-TV used SBP as a time-varying variable; and (3) Model-TV-Lag added a 3-month lag to Model-TV to de-emphasize changes in SBP associated with acute illness. In all the models, pre-dialysis SBP <120 mm Hg was associated with a higher risk of mortality compared with the referent group (140-159 mm Hg); higher pre-dialysis SBP was not associated with higher risk of mortality. In conclusion, we observed a robust association between lower pre-dialysis SBP and higher risk for all-cause and cardiovascular mortality in a well-characterized cohort of prevalent haemodialysis patients. Randomized clinical trials are needed to define optimal blood pressure targets in the haemodialysis population.


Gardnerella vaginalis promotes group B Streptococcus vaginal colonization, enabling ascending uteroplacental infection in pregnant mice.
Group B Streptococcus is a common vaginal bacterium and the leading cause of invasive fetoplacental infections. Group B Streptococcus in the vagina can invade through the cervix to cause ascending uteroplacental infections or can be transmitted to the neonate during vaginal delivery. Some studies have found that women with a "dysbiotic" polymicrobial or Lactobacillus-depleted vaginal microbiota are more likely to harbor group B Streptococcus. Gardnerella vaginalis is often the most abundant bacteria in the vaginas of women with dysbiosis, while being detected at lower levels in most other women, and has been linked with several adverse pregnancy outcomes. Mouse models of group B Streptococcus and Gardnerella vaginalis colonization have been reported but, to the best of our knowledge, the two have not been studied together. The overarching idea driving this study is that certain members of the dysbiotic vaginal microbiota, such as Gardnerella vaginalis, may directly contribute to the increased rate of group B Streptococcus vaginal colonization observed in women with vaginal dysbiosis. We used a mouse model to test the hypothesis that vaginal exposure to Gardnerella vaginalis may facilitate colonization and/or invasive infection of the upper reproductive tract by group B Streptococcus during pregnancy. Timed-pregnant mice were generated using an allogeneic mating strategy with BALB/c males and C57Bl/6 females. Dams were vaginally inoculated at gestational day 14 with group B Streptococcus alone (using a 10-fold lower dose than previously reported models) or coinoculated with group B Streptococcus and Gardnerella vaginalis. Bacterial titers were enumerated in vaginal, uterine horn, and placental tissues at gestational day 17. The presence (Fisher exact tests) and levels (Mann-Whitney U tests) of bacterial titers were compared between mono- and coinoculated dams in each compartment. Relative risks were calculated for outcomes that occurred in both groups. Tissue samples were also examined for evidence of pathophysiology. Inoculation of pregnant mice with 107 group B Streptococcus alone did not result in vaginal colonization or ascending infection. In contrast, coinoculation of group B Streptococcus with Gardnerella vaginalis in pregnant mice resulted in a 10-fold higher risk of group B Streptococcus vaginal colonization (relative risk, 10.31; 95% confidence interval, 2.710-59.04; P=.0006 [Fisher exact test]). Ascending group B Streptococcus infection of the uterus and placenta occurred in approximately 40% of coinoculated animals, whereas none of those receiving group B Streptococcus alone developed uterine or placental infections. Immunofluorescence microscopy revealed group B Streptococcus in both the maternal and fetal sides of the placenta. Histologic inflammation and increased proinflammatory cytokines were evident in the setting of group B Streptococcus placental infection. Interestingly, placentas from dams exposed to group B Streptococcus and Gardnerella vaginalis, but without recoverable vaginal or placental bacteria, displayed distinct histopathologic features and cytokine signatures. These data suggest that Gardnerella vaginalis vaginal exposure can promote group B Streptococcus vaginal colonization, resulting in a greater likelihood of invasive perinatal group B Streptococcus infections. These findings suggest that future clinical studies should examine whether the presence of Gardnerella vaginalis is a risk factor for group B Streptococcus vaginal colonization in women. Because Gardnerella vaginalis can also be present in women without bacterial vaginosis, these findings may be relevant both inside and outside of the context of vaginal dysbiosis.


Depleting neuronal PrP in prion infection prevents disease and reverses spongiosis.
The mechanisms involved in prion neurotoxicity are unclear, and therapies preventing accumulation of PrPSc, the disease-associated form of prion protein (PrP), do not significantly prolong survival in mice with central nervous system prion infection. We found that depleting endogenous neuronal PrPc in mice with established neuroinvasive prion infection reversed early spongiform change and prevented neuronal loss and progression to clinical disease. This occurred despite the accumulation of extraneuronal PrPSc to levels seen in terminally ill wild-type animals. Thus, the propagation of nonneuronal PrPSc is not pathogenic, but arresting the continued conversion of PrPc to PrPSc within neurons during scrapie infection prevents prion neurotoxicity.


GP Trainees experience of learning opportunities and support mechanisms on the GP training programme: a qualitative study.
This article was migrated. The article was marked as recommended. Background: General Practice (GP) is the cornerstone of the NHS and has faced workload and workforce issues for many years. Enabling GP trainees to successfully complete their training and become independent practitioners is crucial to addressing these challenges. There is limited previous research exploring the postgraduate training experience specific to General Practice. Aim: To explore GP Trainees' experience of learning opportunities and support available during the three-year vocational training programme in the Southampton GP Education Unit. Design and Setting: 15 semi structured interviews were undertaken December 2016-2018 with participants recruited from four cohorts. Telephone interviews lasting between 30-60 minutes were audio-recorded and transcribed verbatim; and analysed using inductive thematic analysis. Results and Conclusion: Two themes were identified as affecting the training experience: 'Developing clinical competence' and 'Sources of support'. Trainees perceived developing competencies relevant to General Practice was key. Busier hospital rotations with limited time for formal teaching were viewed less favourably. Trainers and peers were the main sources of support. Despite seeking a broad range of participants, interviewing trainees in difficulty was hard to achieve.


Oxidative stress increases placental and endothelial cell activin A secretion.
Circulating levels of activin A are significantly increased in women with preeclampsia when compared with those with a normal pregnancy. The mechanisms underlying these increased levels are unknown. We undertook these studies to explore whether oxidative stress might be the mechanism. We exposed trophoblast explants, human umbilical vein endothelial cells (HUVECs) and peripheral blood monocytes to oxidative stress in vitro using xanthine/xanthine oxidase (X/XO), measuring activin A and isoprostane in conditioned media and mRNA for activin beta(A) in explants and HUVECs. We also measured isoprostane and activin A in serum from 21 women with preeclampsia and from 20 women with a normal pregnancy. Treatment with X/XO significantly increased 8-isoprostane production from placental explants, HUVECs and monocytes, indicative of oxidative stress, and significantly increased activin A output from placental explants (139.1 +/- 27.4 per mg wet weight vs 322.9 +/- 89.7 pg/ml per mg wet weight, P = 0.02) and from HUVECs (1.2 +/- 0.2 vs 3.2 +/- 1.8 ng/ml, P = 0.04). There was no effect on activin A output from monocytes. X/XO significantly increased beta(A) mRNA in placental explants but not in HUVECs. Maternal plasma levels of 8-isoprostane and activin A were significantly higher in women with preeclampsia when compared with controls (333.8 +/- 70 vs 176.3 +/- 26.2 pg/ml, P = 0.04 and 49.5 +/- 7 vs 13.1 +/- 1.2 ng/ml, P < 0.001 respectively). In the women with preeclampsia, but not in those with a normal pregnancy, circulating levels of 8-isoprostane and activin A were significantly and positively correlated (r(2) = 0.72; P < 0.001). These data suggest that oxidative stress may be one of the mechanisms underlying increased circulating activin A in preeclampsia.


Medicalization, reproductive agency, and the desire for surgical sterilization among low-income women in urban Brazil.
This article draws on data from ethnographic fieldwork in an urban housing project to examine the social context and meanings of surgical sterilization for low-income women in Brazil. Low-income women resort to sterilization because they distrust or are unsatisfied with alternative methods and because it helps them to fulfill the requirements of modern, responsible motherhood. Although sterilization is an option among few alternatives, and one that has subjected women to greater medical management and intervention, I argue that sterilization also represents poor women's active struggle to improve their lives and to resist the burdens placed on them by unequal gender relations. This article contributes to a growing anthropological literature that demonstrates how reproduction has become a central site where social values are constituted and contested, and it details women's diverse responses to the process of medicalization.


Effect of basic and basic/acid modifications on the surface of PVDF membranes for the insertion of TiO2 and its use in environmental applications.
Supporting titanium oxide (TiO2) on polymeric membrane surfaces is a strategy to increase the photocatalytic activity of this material as well as to modify membrane surface with antifouling properties or to develop hybrid processes of water treatment. The chemical characteristics of the polymeric membrane surfaces are a determining factor in the correct impregnation of TiO2 particles. In this work, the titanium oxide was immobilized on polyvinylidene fluoride (PVDF) membrane surface by direct impregnation during the synthesis of the inorganic particles by sol-gel route. The PVDF membranes were previously modified by treatments based on an alkaline attack followed by acid treatment. The final TiO2-modified membranes were characterized by infrared and Raman spectroscopy, as well as by scanning electron microscopy. In addition, the changes on the surface characteristics were determined by contact angle measurements. Finally, the membranes were tested on the photocatalytic degradation of methyl orange (MO). The results obtained indicate that the basic/acid pretreatment allows the generation of active sites in the membrane and that when carrying out the synthesis of TiO2 on the membrane, it can be anchored stably on its surface and through the pores. The microscopies indicate that the structure of the membrane is not compromised by the pretreatment. The amount of TiO2 deposited on the membrane was of 0.1580 ± 0.01773 mg TiO2/cm2 membrane. With this amount of TiO2, a degradation percentage of 98.2% is achieved after 450 min; when the membrane is used for a second cycle, a degradation percentage of 82.0% is obtained, which remains constant for 3 subsequent cycles. This method, which uses the PVDF membrane as a support for TiO2 particles, represents a low-cost and easy-to-prepare insertion procedure, with good degradation percentages, which means that the membrane can be used for subsequent studies in filtration systems in the treatment of effluents from the textile industry.


Geoelectrochemistry-driven alteration of amino acids to derivative organics in carbonaceous chondrite parent bodies.
A long-standing question regarding carbonaceous chondrites (CCs) is how the CCs' organics were sourced and converted before and after the accretion of their parent bodies. Growing evidence shows that amino acid abundances in CCs decrease with an elongated aqueous alteration. However, the underlying chemical processes are unclear. If CCs' parent bodies were water-rock differentiated, pH and redox gradients can drive electrochemical reactions by using H2 as an electron source. Here, we simulate such redox conditions and demonstrate that α-amino acids are electrochemically altered to monoamines and α-hydroxy acids on FeS and NiS catalysts at 25 °C. This conversion is consistent with their enrichment compared to amino acid analogs in heavily altered CCs. Our results thus suggest that H2 can be an important driver for organic evolution in water-rock differentiated CC parent bodies as well as the Solar System icy bodies that might possess similar pH and redox gradients.


Role of interventional radiology in the management of acute gastrointestinal bleeding.
Acute gastrointestinal bleeding (GIB) can lead to significant morbidity and mortality without appropriate treatment. There are numerous causes of acute GIB including but not limited to infection, vascular anomalies, inflammatory diseases, trauma, and malignancy. The diagnostic and therapeutic approach of GIB depends on its location, severity, and etiology. The role of interventional radiology becomes vital in patients whose GIB remains resistant to medical and endoscopic treatment. Radiology offers diagnostic imaging studies and endovascular therapeutic interventions that can be performed promptly and effectively with successful outcomes. Computed tomography angiography and nuclear scintigraphy can localize the source of bleeding and provide essential information for the interventional radiologist to guide therapeutic management with endovascular angiography and transcatheter embolization. This review article provides insight into the essential role of Interventional Radiology in the management of acute GIB.


The clinical laboratory of the future: re-engineering laboratory services.
Re-engineering of the process to deliver laboratory services to patients is essential to meet the pressures for patient-focussed care and cost control. Re-engineering requires that laboratory staff abandon their current thinking and assumptions about organization, roles, procedures, skills and tools to focus on a totally new way of delivering value to the patient. The rewards are high for the organization. The risks must be managed by following a proven approach. The opportunities for personal development are exciting.


Molecular mechanism of immunoglobulin V-region diversification regulated by transcription and RNA metabolism in antigen-driven B cells.
The immune system produces specific antibodies (Ab) against any antigens (Ag) of exogenous and endogenous origins with a diverse repertoire of V-region specificities. The primary V-region repertoire is created by the rearrangement of immunoglobulin (Ig) V-region, D- and J-segments with the insertion of N- and P-sequences during early B cell differentiation. Recent studies revealed that secondary diversification of the IgV-region generated in the peripheral lymphoid organs plays a critical role in the generation of effective Ab production for protection from various pathogens. Naïve B cells that react with Ags initiate proliferation and differentiation in the follicular region and create the germinal centres (GCs), where activation-induced cytidine deaminase (AID)-dependent IgV-region somatic hypermutation (SHM) and class-switch recombination generate high-affinity and class-switched mature Ag-specific B cells. Our studies have discovered a 210-kDa nuclear protein, named GC-associated nuclear protein (GANP) that is up-regulated in GC B cells during the T cell-dependent (TD) immune responses. By studying mice with mutant forms of the ganp gene, we demonstrated that GANP is essential for the generation of high-affinity B cells against TD-Ag by affecting SHM at the IgV-regions. GANP is associated with AID in the cytoplasm and the GANP/AID complex is recruited to the nucleus, specifically, the chromatin, and targeted selectively to the IgV-region gene in B cells. GANP augments the access of AID towards IgV-regions in B cells. Here, we review the role of GANP in acquired immunity through the detailed analysis of the molecular mechanism generating SHM specifically at IgV-regions in B cells.


Outer membrane proteins of Escherichia coli. VII. Evidence that bacteriophage-directed protein 2 functions as a pore.
Protein 1, a major protein of the outer membrane of Escherichia coli, has been shown to be the pore allowing the passage of small hydrophilic solutes across the outer membrane. In E. coli K-12 protein 1 consists of two subspecies, 1a and 1b, whereas in E. coli B it consists of a single species which has an electrophoretic mobility similar to that of 1a. K-12 strains mutant at the ompB locus lack both proteins 1a and 1b and exhibit multiple transport defects, resistance to toxic metal ions, and tolerance to a number of colicins. Mutation at the tolF locus results in the loss of 1a, in less severe transport defects, and more limited colicin tolerance. Mutation at the par locus causes the loss of protein 1b, but no transport defects or colicin tolerance. Lysogeny of E. coli by phage PA-2 results in the production of a new major protein, protein 2. Lysogeny of K-12 ompB mutants resulted in dramatic reversal of the transport defects and restoration of the sensitivity to colicins E2 and E3 but not to other colicins. This was shown to be due to the production of protein 2, since lysogeny by phage mutants lacking the ability to elicit protein 2 production did not show this effect. Thus, protein 2 can function as an effective pore. ompB mutations in E. coli B also resulted in loss of protein 1 and similar multiple transport defects, but these were only partially reversed by phage lysogeny and the resulting production of protein 2. When the ompB region from E. coli B was moved by transduction into an E. coli K-12 background, only small amounts of proteins 1a and 1b were found in the outer membrane. These results indicate that genes governing the synthesis of outer membrane proteins may not function interchangeably between K-12 and B strains, indicating differences in regulation or biosynthesis of these proteins between these strains.


Interferon-gamma potentiates NMDA receptor signaling in spinal dorsal horn neurons via microglia-neuron interaction.
Glia-neuron interactions play an important role in the development of neuropathic pain. Expression of the pro-inflammatory cytokne →cytokine Interferon-gamma (IFNγ) is upregulated in the dorsal horn after peripheral nerve injury, and intrathecal IFNγ administration induces mechanical allodynia in rats. A growing body of evidence suggests that IFNγ might be involved in the mechanisms of neuropathic pain, but its effects on the spinal dorsal horn are unclear. We performed blind whole-cell patch-clamp recording to investigate the effect of IFNγ on postsynaptic glutamate-induced currents in the substantia gelatinosa neurons of spinal cord slices from adult male rats. IFNγ perfusion significantly enhanced the amplitude of NMDA-induced inward currents in substantia gelatinosa neurons, but did not affect AMPA-induced currents. The facilitation of NMDA-induced current by IFNγ was inhibited by bath application of an IFNγ receptor-selective antagonist. Adding the Janus activated kinase inhibitor tofacitinib to the pipette solution did not affect the IFNγ-induced facilitation of NMDA-induced currents. However, the facilitatory effect of IFNγ on NMDA-induced currents was inhibited by perfusion of the microglial inhibitor minocycline. These results suggest that IFNγ binds the microglial IFNγ receptor and enhances NMDA receptor activity in substantia gelatinosa neurons. Next, to identify the effector of signal transmission from microglia to dorsal horn neurons, we added an inhibitor of G proteins, GDP-β-S, to the pipette solution. In a GDP-β-S-containing pipette solution, IFNγ-induced potentiation of the NMDA current was significantly suppressed after 30 min. In addition, IFNγ-induced potentiation of NMDA currents was blocked by application of a selective antagonist of CCR2, and its ligand CCL2 increased NMDA-induced currents. Our findings suggest that IFNγ enhance the amplitude of NMDA-induced inward currents in substantia gelatinosa neurons via microglial IFNγ receptors and CCL2/CCR2 signaling. This mechanism might be partially responsible for the development of persistent neuropathic pain.


Tailored multilobar disconnective epilepsy surgery in the posterior quadrant.
Surgical treatment of drug-resistant epilepsy originating from the posterior quadrant (PQ) of the brain often requires large multilobar resections, and disconnective techniques have been advocated to limit the risks associated with extensive tissue removal. Few previous studies have described a tailored temporoparietooccipital (TPO) disconnective approach; only small series with short postoperative follow-ups have been reported. The aim of the present study was to present a tailored approach to multilobar PQ disconnections (MPQDs) for epilepsy and to provide details about selection of patients, presurgical investigations, surgical technique, treatment safety profile, and seizure and cognitive outcome in a large, single-center series of patients with a long-term follow-up. In this retrospective longitudinal study, the authors searched their prospectively collected database for patients who underwent MPQD for drug-resistant epilepsy in the period of 2005-2017. Tailored MPQDs were a posteriori grouped as follows: type I (classic full TPO disconnection), type II (partial TPO disconnection), type III (full temporooccipital [TO] disconnection), and type IV (partial TO disconnection), according to the disconnection plane in the occipitoparietal area. A bivariate statistical analysis was carried out to identify possible predictors of seizure outcome (Engel class I vs classes II-IV) among several presurgical, surgical, and postsurgical variables. Preoperative and postoperative cognitive profiles were also collected and evaluated. Forty-two consecutive patients (29 males, 24 children) met the inclusion criteria. According to the presurgical evaluation (including stereo-electroencephalography in 13 cases), 12 (28.6%), 24 (57.1%), 2 (4.8%), and 4 (9.5%) patients received a type I, II, III, or IV MPQD, respectively. After a mean follow-up of 80.6 months, 76.2% patients were in Engel class I at last contact; at 6 months and 2 and 5 years postoperatively, Engel class I was recorded in 80.9%, 74.5%, and 73.5% of cases, respectively. Factors significantly associated with seizure freedom were the occipital pattern of seizure semiology and the absence of bilateral interictal epileptiform abnormalities at the EEG (p = 0.02). Severe complications occurred in 4.8% of the patients. The available neuropsychological data revealed postsurgical improvement in verbal domains, whereas nonunivocal outcomes were recorded in the other functions. The presented data indicate that the use of careful anatomo-electro-clinical criteria in the presurgical evaluation allows for customizing the extent of surgical disconnections in PQ epilepsies, with excellent results on seizures and an acceptable safety profile.


AdipoRon reduces cisplatin-induced ototoxicity in hair cells:possible relation to the regulation of mitochondrial biogenesis.
AdipoRon (AR) can exert antidiabetic and anti-inflammatory effects by maintaining mitochondrial structure and function. The present study was designed to explore whether AR protects the auditory cells from cisplatin-induced damage and, if so, to probe the possible mechanisms underlying its action on this type of cells. Cell viability and apoptosis in House Ear Institute-Organization of Corti 1 (HEI-OC1 cells) and mouse cochlea hair cells (HCs) were detected by CCK8 and immunofluorescence. The expressions of apoptosis-related proteins (cleaved caspase-3 and Bcl-2), adiponectin receptor 1 (AdipoR 1) and the key factors relevant to mitochondrial biogenesis(SIRT1 and TFAM)were determined by Western blot and immunofluorescence. Changes in apoptotic rate and expression of SIRT1 and TFAM after silencing of AdipoR 1 (AdipoR 1-siRNA) in HEI-OC1 cells were measured by flow cytometry and Western blot. The levels of reactive oxygen species (ROS) were evaluated by MitoSox red staining. We found that 30 μM cisplatin exposure induced severe cellular damage, which resulted from activation of the mitochondrial apoptotic pathway. Cisplatin decreased the expression of AdipoR 1, SIRT1, and TFAM proteins, leading to impaired mitochondrial biogenesis and increased mitochondrial ROS production. 10 μM AR pre-treatment enhanced mitochondrial biogenesis, decreased mitochondrial ROS levels, alleviated imbalances in the mitochondrial apoptotic pathway, thus reducing cisplatin-induced apoptosis. Taken together, this work reveals that AR exerts anti-apoptotic effects, possibly via regulating mitochondrial biogenesis and function. Interestingly, AR might possess the promising potential to be a novel drug for the prevention and/ or treatment of cisplatin-induced ototoxicity.


Gametocytogenesis in malaria parasite: commitment, development and regulation.
Malaria parasites have evolved a complicated life cycle alternating between two hosts. Gametocytes are produced in the vertebrate hosts and are obligatory for natural transmission of the parasites through mosquito vectors. The mechanism of sexual development in Plasmodium has been the focus of extensive studies. In the postgenomic era, the advent of genome-wide analytical tools and genetic manipulation technology has enabled rapid advancement of our knowledge in this area. Patterns of gene expression during sexual development, molecular distinction of the two sexes, and mechanisms underlying subsequent formation of gametes and their fertilization have been progressively elucidated. However, the triggers and mechanism of sexual development remain largely unknown. This article provides an update of our understanding of the molecular and cellular events associated with the decision for commitment to sexual development and regulation of gene expression during gametocytogenesis. Insights into the molecular mechanisms of gametocyte development are essential for designing proper control strategies for interruption of malaria transmission and ultimate elimination.


Radiolucent lines around knee arthroplasty components : a narrative review.
Aseptic loosening of total knee arthroplasty (TKA) components is one of the frequent reasons for early revision together with infection and instability. Aseptic loosening is usually preceded by the observation of radiolucent lines (RLL) on radiographs. Radiolucent lines have conventionally been considered a sign of osteolysis due to particles disease of either polyethylene or cement wear. However, RLL can be observed quite early after TKA, way before wear and osteolysis can even occur. Immediate postoperative RLL are secondary to surgical technique with either inadequate cement penetration in sclerotic bone, insufficient preparation of the bone or malpositioning of the component relative to the bone cuts. This type of RLL can be observed radiologically but remains often without clinical symptoms. Early development of RLL, on an initially satisfying radiograph, is secondary to changes to the cement-bone interface. These are most often related to micromotion because of constraint, malalignment, remaining mechanical deformity, erroneous bone cuts or osteoporosis. This type of RLL are observed progressively on follow-up radiographs and can be accompanied by pain complaints despite of initial good outcome. Young age, male sex or osteoporotic bones often found in elderly females, are all risk factors. A special form of aseptic loosening is tibial debonding that has been observed for different types of implants and different types of cement. It occurs at the cement-implant interface with cement remaining well attached to the trabecular bone. Probably it is a lack of cement adhesion between the high viscosity cement and the component. Revision is proposed upon diagnosis to avoid component's displacement with secondary destruction of the proximal tibial bone. Finally, RLL can develop over time secondary to polyethylene wear. These lines appear because of osteolysis and bone loss and will lead at the end to aseptic loosening of the components. Symptoms are related to failure of the implant-bone construct. Radiolucent lines without clinical symptoms should be analysed according to their potential reason of development and followed up closely with adequate radiological techniques. If symptoms develop or radiological imaging objectivizes failure and component mobility, revision knee arthroplasty might be necessary.


Restoration of bacterial killing activity of human respiratory cystic fibrosis cells through cationic vector-mediated cystic fibrosis transmembrane conductance regulator gene transfer.
In vitro and in vivo studies have demonstrated that gene transfer of the CFTR (cystic fibrosis transmembrane conductance regulator) cDNA into human respiratory cells through nonviral vectors can occur safely and can be done repeatedly. Although functional evaluation of CFTR in cystic fibrosis (CF) patients enrolled in phase I clinical trials using cationic liposomes has shown a partial correction of nasal potential difference, a biological assay indicating a therapeutic relevance of CFTR gene transfer is still missing. Our aims were to study the induction of killing activity toward Pseudomonas aeruginosa (PA) in CF cells by cationic vector-mediated CFTR gene transfer and to use this assay as a therapeutic end point. Luciferase expression and GFP FACS analysis were used to evaluate the optimal vector and the efficiency of gene transfer into non-CF human respiratory cells growing from nasal polyp explants at the air-liquid interface. To prove that transgenic CFTR was expressed in CF cell cultures under the same experimental conditions, a specific RT-PCR was performed. Challenge of the outgrowths with a known amount of PA showed a bacterial clearance activity by non-CF respiratory cells, while in the case of CF cells it even resulted in bacterial growth. Cationic vector-mediated CFTR cDNA determined the recovery of bacterial clearance activity only under those conditions yielding 5% or more of GFP-positive cells. The results shown in this study might be helpful in considering cationic vectors as therapeutic nonviral vectors for transferring CFTR into human CF respiratory cells, as well as for restoring the bacterial killing activity defective in cystic fibrosis.


Relations among daily stressors, childhood maltreatment, and sleep in college students.
This study aimed to assess (a) the within- and between-person associations between daily stress and sleep; (b) the relation between childhood maltreatment and sleep; and (c) whether the relation between stress and sleep was moderated by the extent of childhood maltreatment among college students. Participants (N = 181) comprised the active control group in a previous intervention study. Participants completed a self-report measure of childhood maltreatment and 14 daily self-report measures of stressor exposure and severity (evenings) and 6 sleep measures (e.g., quality, duration; mornings). Experiencing more daily stressors than usual (within-person relation) was significantly associated with delayed sleep latency (i.e., time falling asleep). Greater daily stressor severity was also significantly associated with lower sleep efficiency within persons. Participants who reported more stressors in general had shorter sleep duration, poorer sleep quality, and less restful sleep (between-person relations). Students who reported more childhood maltreatment also reported significantly lower quality sleep and feeling less rested upon awakening. Childhood maltreatment did not moderate the within-person association between daily stress and sleep. Unexpectedly, at the between-person level, maltreatment moderated the association between stressors and stressor severity and several sleep parameters (e.g., efficiency and latency) such that there was a weaker relation between stress and sleep among those with more maltreatment. Interventions on campus could aim to reduce stress and improve sleep. Additional awareness of the prevalence of maltreatment and how it may be related to sleep also appears warranted. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Diversity and Distribution of Cryptic Species of the Bemisia tabaci (Hemiptera: Aleyrodidae) complex in Pakistan.
Bemisia tabaci (Gennadius; Hempitera: Aleyrodidae) is considered to be a cryptic (sibling) species complex, the members of which exhibit morphological invariability while being genetically and behaviorally distinct. Members of the complex are agricultural pests that cause direct damage by feeding on plants, and indirectly by transmitting viruses that cause diseases leading to reduced crop yield and quality. In Pakistan, cotton leaf curl disease, caused by multiple begomovirus species, is the most economically important viral disease of cotton. In the study outlined here, the diversity and geographic distribution of B. tabaci cryptic species was investigated by analyzing a taxonomically informative fragment of the mitochondrial cytochrome c oxidase 1 gene (mtCOI-3'). The mtCOI-3' sequence was determined for 285 adult whiteflies and found to represent six cryptic species, the most numerous being Asia II-1 and Middle East Asia Minor 1 (MEAM-1), the later also referred to as the B-biotype, which was previously thought to be confined to Sindh province but herein, was also found to be present in the Punjab province. The endemic Asia I was restricted to Sindh province, while an individual in the Asia II-8 was identified in Pakistan for the first time. Also for the first time, samples were collected from northwestern Pakistan and Asia II-1 was identified. Results indicate that in Pakistan the overall diversity of B. tabaci cryptic species is high and, based on comparisons with findings from previous studies, the distribution is dynamic.


The factors affecting neonatal presentations to the pediatric emergency department.
A pediatric emergency department (PED) may be utilized by neonates for nonurgent complaints. Various factors, such as primiparity, maternal age, early postnatal discharge, race, income, and maternal and paternal educational levels, have been reported to affect the acuity of neonatal emergency department utilization. To determine the characteristics of PED visits by neonates (infants ≤ 28 days of age) and to evaluate the factors affecting the acuity of these visits. We prospectively collected the data of neonates who were admitted to the PED of a tertiary university hospital within a 6-month period. Presenting problems were classified as acute if diagnostic tests were requested or the patient was hospitalized, unless the final diagnosis was "normal newborn." Over this period, 28,389 children (0-18 years of age) visited the PED, of which 531 were newborns (1.9%). The mean age was 14.1 ± 8.3 days, with a slight predominance of males (57.3%). The chief complaints were jaundice (23.4%), irritability (9.5%), and vomiting (7.1%), and the most common diagnoses were normal newborn (33.9%), indirect hyperbilirubinemia (13.2%), and colic (5.8%). Acute visits were 55.7% of the total visits. Premature infants, infants of multiparous mothers, infants of older mothers (≥25 years), and physician-referred infants were more likely to present with acute problems (p values were 0.001, 0.013, 0.006, and <0.001, respectively). The results suggest that there may be a relationship between nonacute neonatal visits to a PED and insufficient knowledge of the caretaker on newborn care. Thus, more detailed education and early postnatal support programs regarding newborn care may help to decrease nonacute PED visits by neonates.


Properties of some norvaline-resistant mutants of Bacillus subtilis.
DL-Norvaline inhibits growth of wild-type Bacillus subtilis. A number of mutants resistant to growth inhibition by this analogue were isolated and studied. Cross-feeding experiments and paper chromatography of culture supernatants indicated that the mutants excreted leucine and possibly valine and glutamate. Enzymic analysis indicated that the mutants were derepressed for acetohydroxy-acid synthetase and alpha-isopropylmalate synthetase; however, no derepression of threonine deaminase, dihydroxyacid dehydrase or transaminase B was observed.


Development of novel formulations to enhance in vivo transdermal permeation of tocopherol.
Tocopherol represents a big challenge for transdermal permeation owing to its extreme hydrophobicity and large molecular mass. The aim of the present study was to develop alpha-tocopherol (T) topical formulations and evaluate their ex vivo and in vivo permeation. Franz diffusion cells were used for ex vivo permeation, and neonatal rats were used for in vivo permeation. Seven gel formulations and 21 liquid formulations were investigated for physical stability, viscosity and permeation of T. Analysis of T was performed by a validated HPLC method using a UV detector. The ex vivo permeation from gel and emulsion formulations was very poor (0.001-0.015%). Highest permeation was observed from monophasic liquid formulations containing dimethyl sulfoxide (DMSO), tocopheryl polyethylene glycols (TPGs), propylene glycol, ethanol and 9.5% T. The in vivo results demonstrated higher retention in the epidermis compared to subcutaneous tissues, 1377 and 1.13 μg g⁻¹, respectively. Increasing T concentration from 4.8 to 9.5% did not increase the amount permeated or % of T retained. It was concluded that simple solutions of T in the presence of DMSO and TPGs were more promising systems for effective transdermal permeation compared to gel, emulsion or oleaginous systems.


A novel application of teleneurology: robotic telepresence in supervision of neurology trainees.
Telemedicine is used for specialist consultation, patient evaluation, and remote monitoring. We propose that telemedicine has a valuable role in medical education through facilitating faculty supervision of trainees. We surveyed resident physician trainees and faculty at a single institution who provided service for acute neurologic consultations over 29 months. The survey compared experiences using the supervisory methods of telephone, robotic telepresence (RTP), and in-person interaction. Participants answered 10 questions addressing their experience relevant to faculty-trainee interaction quality and clinical supervision, achieved with the three supervisory methods under study. Surveys were received from 20 of 36 trainees (56%) and 8 of 10 faculty members (80%). More than 90% of trainees and faculty strongly agreed that in-person communication was satisfactory for patient data review, problem synthesis, and refinement of the trainees' impressions, with RTP ranking a close second. Moreover, 38% of faculty and 70% of trainees reported the telephone method as unsatisfactory for reacquisition of patient history, data synthesis, and direct observation of trainees' clinical skills. RTP provided more than 90% of trainees with sufficient staff support and facilitated trainee autonomy. Overall, 85% of both trainees and faculty preferred in-person supervision most, with RTP ranked second and telephone being least favored. Although in-person faculty supervision of trainees' management of neurologic emergencies is always preferred, it is not always practical. This survey highlights the valuable role of RTP in graduate medical education and RTP as preferred over telephone as an adjunctive supervision tool in residency.


LGR5 promotes survival in human colorectal adenoma cells and is upregulated by PGE2: implications for targeting adenoma stem cells with NSAIDs.
Cyclooxygenase-2 is overexpressed in the majority of colorectal tumours leading to elevated levels of prostaglandin E2 (PGE2), promoting many hallmarks of cancer. Importantly, PGE2 is reported to enhance Wnt/β-catenin signalling in colorectal carcinoma cells and in normal haematopoietic stem cells where it promotes stem cell function. Although Wnt signalling plays a crucial role in intestinal stem cells, the relationship between PGE2 and intestinal stem cells is unclear. Given that the key intestinal cancer stem cell marker LGR5 (leucine-rich G-protein coupled receptor 5) is a Wnt target and PGE2 enhances Wnt signalling, the focus of this study was to investigate whether PGE2 regulated LGR5 expression in colorectal adenoma cells and whether LGR5 was important for tumour cell survival. PGE2 upregulated LGR5 protein in adenoma (RG/C2) and carcinoma (DLD-1) cell lines. LGR5 knockdown induced cell death in RG/C2 and AA/C1 adenoma cells, suggesting that LGR5 has an important survival-promoting role in adenoma cells. Indeed, we detected LGR5 protein expression in 4 of 4 human adenoma cell lines. Furthermore, LGR5 small interfering RNA inhibited the survival-promoting effects of PGE2 in RG/C2, suggesting that PGE2 promotes adenoma cell survival, at least in part, by increasing LGR5 expression. These studies, therefore, show the first link between PGE2 and LGR5 in human colorectal adenoma and carcinoma cells and demonstrate a survival-promoting role of LGR5. As non-steroidal anti-inflammatory drugs (NSAIDs) cause adenomas to regress in FAP patients, these studies could have important implications for the mechanism by which NSAIDs are chemopreventive, as lowering PGE2 levels could reduce LGR5 expression and survival of LGR5(+) adenoma stem cells.


Food Workers as a Reservoir of Extended-Spectrum-Cephalosporin-Resistant Salmonella Strains in Japan.
Dissemination of extended-spectrum-cephalosporin (ESC)-resistant Salmonella, especially extended-spectrum-β-lactamase (ESBL)-producing Salmonella, is a concern worldwide. Here, we assessed Salmonella carriage by food workers in Japan to clarify the prevalence of ESC-resistant Salmonella harboring blaCTX-M We then characterized the genetic features, such as transposable elements, of blaCTX-M-harboring plasmids using whole-genome sequencing. A total of 145,220 stool samples were collected from food workers, including cooks and servers from several restaurants, as well as food factory workers, from January to October 2017. Isolated salmonellae were subjected to antimicrobial susceptibility testing (disk diffusion method), and whole-genome sequencing was performed for Salmonella strains harboring blaCTX-M Overall, 164 Salmonella isolates (0.113%) were recovered from 164 samples, from which we estimated that at least 0.113% (95% confidence interval [CI]: 0.096 to 0.132%) of food workers may carry Salmonella Based on this estimation, 3,473 (95% CI = 2,962 to 4,047) individuals among the 3,075,330 Japanese food workers are likely to carry Salmonella Of the 158 culturable isolates, seven showed resistance to ESCs: three isolates harbored blaCMY-2 and produced AmpC β-lactamase, while four ESBL-producing isolates harbored blaCTX-M-14 (n = 1, Salmonella enterica serovar Senftenberg) or blaCTX-M-15 (n = 3, S. enterica serovar Haardt). blaCTX-M-15 was chromosomally located in the S Haardt isolates, which also contained ISEcp1, while the S Senftenberg isolate contained an IncFIA(HI1)/IncHI1A/IncHI1B(R27) hybrid plasmid carrying blaCTX-M-14 along with ISEcp1 This study indicates that food workers may be a reservoir of ESBL-producing Salmonella and associated genes. Thus, these workers may contribute to the spread of blaCTX-M via plasmids or mobile genetic elements such as ISEcp1IMPORTANCE Antimicrobial-resistant Salmonella bacteria arise in farm environments through imprudent use of antimicrobials. Subsequently, these antimicrobial-resistant strains, such as extended-spectrum-β-lactamase (ESBL)-producing Salmonella, may be transmitted to humans via food animal-derived products. Here, we examined Salmonella carriage among food handlers in Japan. Overall, 164 of 145,220 fecal samples (0.113%) were positive for Salmonella Among the 158 tested isolates, four were identified as ESBL-producing isolates carrying ESBL determinants blaCTX-M-15 or blaCTX-M-14 In all cases, the genes coexisted with ISEcp1, regardless of whether they were located on the chromosome or on a plasmid. Our findings suggest that food workers may be a reservoir of ESBL-producing strains and could contribute to the spread of resistance genes from farm-derived Salmonella to other bacterial species present in the human gut.


Reshaping students' attitudes toward individuals with mental illness through a clinical nursing clerkship.
The purpose of this study was to evaluate the degree to which a psychiatric clinical clerkship alters nursing students' attitudes toward individuals with mental illness. The goal of the 4-week clerkship is to provide students with the knowledge, skills, and professional attitudes that will enable them to care for individuals with mental illness in different health care settings. A pencil-and-paper questionnaire was administered to 126 third-year students before and after the clerkship. Students were presented with four hypothetical patients with schizophrenia and were asked to what extent the patient was responsible for his condition, the emotions students felt, and whether students were willing to care for the patient or to segregate him in the hospital. After the clinical clerkship, students became more compassionate and less frightened by psychiatric patients, were more willing to care for individuals with mental illness, and expressed less need to segregate them from the community. In addition, in accord with professional attitudes, students became aware of their own attitudes (the responsibility attributed to patients) and their emotional responses, but these were no longer associated with reluctance to provide care.


[The effect of age on short-wavelength sensitive cone electroretinograms and long-and middle-wavelength sensitive cone electroretinograms].
We studied the effect of age on short-wavelength sensitive cone electroretinogram (S-cone ERG) and long-and middle-wavelength sensitive cone ERG (LM-cone ERG) using a contact lens electrode with a built-in light-emitting diode. We recorded ERGs in 31 pseudophakic subjects to avoid the effect of yellowing in human crystalline lens. The intensities of our stimuli were on the asymptote of the intensity-response curve. We performed linear regression analysis against age on the S-cone ERG b-wave (S-b), and the LM-cone ERG a-, b- and d-waves (LM-a, LM-b, LM-d). We found significant age-dependent reduction in the amplitude of S-b, LM-a, and LM-d, significant prolongation in the peak time of LM-b and LM-d, and significant increase in the b/a ratio of the LM-cone ERG, but no significant age correlation between the peak time of S-b and LM-a and the amplitude of the b-wave. Our results provide evidence that age-related changes in S- and LM-cone systems begin in the twenties in humans. Furthermore, a significant increase of the b/a ratio suggests that off-bipolar cells are more vulnerable to aging than on-bipolar cells.


Click-evoked responses in vestibular afferents in rats.
Sound activates not only the cochlea but also the vestibular end organs. Research on this phenomenon led to the discovery of the sound-evoked vestibular myogenic potentials recorded from the sternocleidomastoid muscles (cervical VEMP, or cVEMP). Since the cVEMP offers simplicity and the ability to stimulate each labyrinth separately, its values as a test of human vestibular function are widely recognized. Currently, the cVEMP is interpreted as a test of saccule function based on the assumption that clicks primarily activate the saccule. However, sound activation of vestibular end organs other than the saccule has been reported. To provide the neural basis for interpreting clinical VEMP testing, we employed the broadband clicks used in clinical VEMP testing to examine the sound-evoked responses in a large sample of vestibular afferents in Sprague-Dawley rats. Recordings were made from 924 vestibular afferents from 106 rats: 255 from the anterior canal (AC), 202 from the horizontal canal (HC), 177 from the posterior canal (PC), 207 from the superior vestibular nerve otolith (SO), and 83 from the inferior nerve otolith (IO). Sound sensitivity of each afferent was quantified by computing the cumulative probability of evoking a spike (CPE). We found that clicks activated irregular afferents (normalized coefficient of variation of interspike intervals >0.2) from both the otoliths (81%) and the canals (43%). The order of end organ sound sensitivity was SO = IO > AC > HC > PC. Since the sternocleidomastoid motoneurons receive inputs from both the otoliths and the canals, these results provide evidence of a possible contribution from both of them to the click-evoked cVEMP.


Progesterone receptor-dependent regulation of genes in the oviducts of female mice.
Oviducts play a critical role in gamete and embryo transport, as well as supporting early embryo development. Progesterone receptor (PGR) is a transcription factor highly expressed in oviductal cells, while its activating ligand, progesterone, surges to peak levels as ovulation approaches. Progesterone is known to regulate oviduct cilia beating and muscular contractions in vitro, but how PGR may mediate this in vivo is poorly understood. We used PGR null mice to identify genes potentially regulated by PGR in the oviducts during the periovulatory period. Histologically, oviducts from PGR null mice showed no gross structural or morphological defects compared with normal littermates. However, microarray analysis of oviducts at 8 h posthuman chorionic gonadotropin revealed >1,000 PGR-dependent genes. Using reverse-transcription polymerase chain reaction (RT-PCR) we selected 10 genes for validation based on their potential roles in oocyte/embryo transport and support. Eight genes were confirmed to be downregulated (Adamts1, Itga8, Edn3, Prlr, Ptgfr, Des, Myocd, and Actg2) and one upregulated (Agtr2) in PGR null oviducts. Expression of these genes was also assessed in oviducts of naturally cycling mice during ovulation and day 1 and day 4 of pregnancy. Adamts1, Itga8, Edn3, Prlr, and Ptgfr were significantly upregulated in oviducts at ovulation/mating. However, most genes showed basal levels of expression at other times. The exceptions were Prlr and Ptgfr, which showed pulsatile increases on day 1 and/or day 4 of pregnancy. This is the first, comprehensive study to elucidate putative PGR-regulated genes in the oviduct and reveals key downstream targets potentially mediating oocyte and embryo transport.


The Occurrence, Causes and Perceived Performance Effects of Breast Injuries in Elite Female Athletes.
Female breasts are vulnerable to direct blows or frictional injuries during sport; however, little research has investigated breast injuries experienced by female athletes. This study aimed to investigate the occurrence, causes and perceived performance effects of breast injuries in elite female athletes across a wide range of sports. A custom-designed survey was distributed to female athletes aged over 18 years who were competing nationally or internationally in their chosen sport. The survey included questions about breast injuries sustained during training and competition and any perceived performance effects of these injuries. 504 elite female athletes from 46 different sports completed the survey. 36% of participants (n = 182) reported experiencing breast injuries and 21% (n = 37) perceived that their breast injury negatively affected their performance. Contact breast injuries were reported by significantly more athletes involved in contact or combat sports and by athletes with larger breasts or a higher body mass index. Frictional breast injuries were reported by significantly more older athletes or those with larger breasts. Less than 10% of participants who experienced breast injuries reported their injury to a coach or medical professional and only half used any prevention strategies. Athletes, coaches and medical professionals associated with women's sport need to be made aware of the occurrence and potential negative effects of breast injuries. It is critical to normalise conversations around breast health so that athletes can be encouraged to report and, when necessary, receive treatment for breast injuries. Further research is also required to better understand factors that affect breast injuries in sport in order to develop evidence-based breast injury prevention strategies.


Gene targeting in murine embryonic stem cells: introduction of specific alterations into the mammalian genome.
The ability to create targeted mutations in the mouse will have an impact on many areas of research in mammalian biology. Mutations are generated in embryonic stem (ES) cells by homologous recombination between exogenously added DNA and the endogenous chromosomal sequences. These cells are then used to generate chimeric intermediates that pass the mutant allele through the germ line, initiating a strain of mice that carry the desired mutation. This review focuses on the selection of a starting ES cell line, introduction of DNA into ES cells, construction of gene targeting vectors, and selection/enrichment schemes for the isolation of targeted cell lines. The generation of mice that carry the targeted allele is briefly outlined.


Phenotype of Children with QT Prolongation Identified Using an Institution-Wide QT Alert System.
QT prolongation is an independent risk factor for cardiovascular mortality in adults. However, there is little information available on pediatric patients with QT prolongation and their outcomes. Herein, we evaluated the prevalence of QT prolongation in pediatric patients identified by an institution-wide QT alert system, and the spectrum of their phenotype. Patients with documented QT prolongation on an ECG obtained between November 2010 and June 2011 were included. There were 1303 pediatric ECGs, and 68 children had electrographically isolated QT prolongation. Comprehensive review of medical records was performed with particular attention to QT-prolonging clinical, laboratory, and medication data, which were summarized into a pro-QTc score. Overall, 68 (5 %) pediatric patients had isolated QT prolongation. The mean age of this pediatric cohort was 9 ± 6 years, and the average QTc was 494 ± 42 ms. All children had 1 or more QT-prolonging risk factor(s), most commonly QT-prolonging medications. One patient was identified with congenital long QT syndrome (LQTS), which was not previously diagnosed. In one-year follow-up, only one pediatric death (non-cardiac) occurred (1.5 %). Potentially QT-offending/pro-arrhythmic medications were changed in 80 % of pediatric patients after the physician received the QT alert. Children with QT prolongation had very low mortality and minimal polypharmacy. Still, medications and other modifiable conditions were the most common causes of QT prolongation. Children with a prolonged QTc should be evaluated for modifiable QT-prolonging factors. However, if no risk factors are present or the QTc does not attenuate after risk factor modification/removal, the child should be evaluated for congenital LQTS.


[The becoming of public medicine in the second half of XVIII-first half of XIX centuries. Report II. The development of public systems of training of medical manpower and charity provision to socially unprotected groups of population].
The present report considers the history of becoming of concept of medical police in second half of XVIII century. This concept became one of the most important instruments of public management in Austria, France, Prussia and Russia. Two directions of activity of public authorities in the area of implementation of medical police are discussed i.e. control of frauds and development of public systems of training of medical manpower and charity provision to socially unprotected groups of population. The historiographical data is presented concerning the development of public systems of training of medical manpower, reform of university medical education, implementation of hospital reform.


[A mobile genetic element on a Bordetella pertussis chromosome stimulates cointegrate formation in Escherichia coli strains].
This work continues a series of reports devoted to the study of a repeated sequence isolated from the Bordetella pertussis chromosome (RSBP--Repeated Sequence of Bordetella Pertussis). The RSBP was earlier demonstrated to have a structure similar to IS elements and exhibit some properties of mobile genetic elements. The results of this work demonstrate the ability of this novel genetic element to stimulate the formation of cointegrates between independent replicons. We have studied the structure of five cointegrates formed by integration of a resident RSBP-containing plasmid into three different sites of a recipient replicon. The studied cointegrates are capable of resolution with the release of a resident plasmid independent of the host cell RecA function.


Wood Anatomy of Douglas-Fir in Eastern Arizona and Its Relationship With Pacific Basin Climate.
Dendroclimatic reconstructions, which are a well-known tool for extending records of climatic variability, have recently been expanded by using wood anatomical parameters. However, the relationships between wood cellular structures and large-scale climatic patterns, such as El Niño-Southern Oscillation (ENSO) and the Pacific Decadal Oscillation (PDO), are still not completely understood, hindering the potential for wood anatomy as a paleoclimatic proxy. To better understand the teleconnection between regional and local climate processes in the western United States, our main objective was to assess the value of these emerging tree-ring parameters for reconstructing climate dynamics. Using Confocal Laser Scanning Microscopy, we measured cell lumen diameter and cell wall thickness (CWT) for the period 1966 to 2015 in five Douglas-firs [Pseudotsuga menziesii (Mirb.) Franco] from two sites in eastern Arizona (United States). Dendroclimatic analysis was performed using chronologies developed for 10 equally distributed sectors of the ring and daily climatic records to identify the strongest climatic signal for each sector. We found that lumen diameter in the first ring sector was sensitive to previous fall-winter temperature (September 25th to January 23rd), while a precipitation signal (October 27th to February 13th) persisted for the entire first half of the ring. The lack of synchronous patterns between trees for CWT prevented conducting meaningful climate-response analysis for that anatomical parameter. Time series of lumen diameter showed an anti-phase relationship with the Southern Oscillation Index (a proxy for ENSO) at 10 to 14year periodicity and particularly in 1980-2005, suggesting that chronologies of wood anatomical parameters respond to multidecadal variability of regional climatic modes. Our findings demonstrate the potential of cell structural characteristics of southwestern United States conifers for reconstructing past climatic variability, while also improving our understanding of how large-scale ocean-atmosphere interactions impact local hydroclimatic patterns.


MicroRNA profiling reveals new aspects of HIV neurodegeneration: caspase-6 regulates astrocyte survival.
MicroRNAs (miRNAs) are small noncoding RNA molecules, which are known to regulate gene expression in physiological and pathological conditions. miRNA profiling was performed using brain tissue from patients with HIV encephalitis (HIVE), a neuroinflammatory/degenerative disorder caused by HIV infection of the brain. Microarray analysis showed differential expression of multiple miRNAs in HIVE compared to control brains. Target prediction and gene ontology enrichment analysis disclosed targeting of several gene families/biological processes by differentially expressed miRNAs (DEMs), with cell death-related genes, including caspase-6, showing a bias toward down-regulated DEMs. Consistent with the miRNA data, HIVE brains exhibited higher levels of caspase-6 transcripts compared with control patients. Immunohistochemical analysis showed localization of the cleaved form of caspase-6 in astrocytes in HIVE brain sections. Exposure of cultured human primary astrocytes to HIV viral protein R (Vpr) induced p53 up-regulation, loss of mitochondrial membrane potential, and caspase-6 activation followed by cell injury. Transgenic mice, expressing Vpr in microglial cells, demonstrated astrocyte apoptosis in brain, which was associated with caspase-6 activation and neurobehavioral abnormalities. Overall, these data point to previously unrecognized alterations in miRNA profile in the brain during HIV infection, which contribute to cell death through dysregulation of cell death machinery.


Acheulean technological behaviour in the Middle Pleistocene landscape of Mieso (East-Central Ethiopia).
The Mieso valley is a new paleoanthropological sequence located in East-Central Ethiopia. It contains Middle and Upper Pleistocene deposits with fossil and lithic assemblages in stratified deposits. This paper introduces the Middle Pleistocene archaeological sequence, attributed to the late Acheulean. Low density clusters of artefacts suggest short-term use of the landscape by Acheulean hominins. In Mieso 31, one of the excavated assemblages, refit sets indicate fragmentation of the reduction sequences and enable study of the initial stages of biface manufacture. Mieso 7, also a stratified site, is primarily characterized by a small concentration of standardized cleavers, and portrays another dimension of Acheulean technology, that related to final stages of use and discard of large cutting tools. Available radiometric dates place the Mieso Acheulean around 212 ka (thousands of years) ago, which would make this sequence among the latest evidence of the Acheulean in East Africa, in a time span when the Middle Stone Age is already documented in the region.


Persistent sleep-disordered breathing independently contributes to metabolic syndrome in prepubertal children.
Obstructive sleep apnea (OSA) is a risk factor for metabolic syndrome (MetS) in adults, but its association in prepubertal children is still questionable due to the relatively limited cardiometabolic data available and the phenotypic heterogeneity. To identify the role of OSA as a potential mediator of MetS in prepubertal children. A total of 255 prepubertal children from the Childhood Adenotonsillectomy Trial were included, with standardized measurements taken before OSA treatment and 7 months later. MetS was defined if three or more of the following criteria were present: adiposity, high blood pressure, elevated glycemia, and dyslipidemia. A causal mediation analysis was conducted to assess the effect of OSA treatment on MetS. OSA treatment significantly impacted MetS, with the apnea-hypopnea index emerging as mediator (p = .02). This mediation role was not detected for any of the individual risk factors that define MetS. We further found that the relationship between MetS and OSA is ascribable to respiratory disturbance caused by the apnea episodes, while systemic inflammation as measured by C-reactive protein, is mediated by desaturation events and fragmented sleep. In terms of evolution, patients with MetS were significantly more likely to recover after OSA treatment (odds ratio = 2.56, 95% confidence interval [CI] 1.20-5.46; risk ratio = 2.06, 95% CI 1.19-3.54) than the opposite, patients without MetS to develop it. The findings point to a causal role of OSA in the development of metabolic dysfunction, suggesting that persistent OSA may increase the risk of MetS in prepubertal children. This mediation role implies a need for developing screening for MetS in children presenting OSA symptoms.


Pickering emulsion templated layer-by-layer assembly for making microcapsules.
Pickering emulsions stabilized by poly(sodium styrenesulfonate) (PSS) surface-modified LUDOX CL particles were used as templates for the layer-by-layer (LbL) deposition of polyelectrolytes and charged nanoparticles to form composite shells. The microcapsules resulting from repeated LbL coating with poly(diallyldimethylammonium chloride) (PDADMAC) and PSS had porous walls due to the loose arrangement of the original nanoparticle aggregates at the oil-water interface, leading to significant microcapsule rupture and low encapsulation efficiency. Microcapsules formed by coating with PDADMAC and anionic LUDOX HS nanoparticles led to dense walls and stronger microcapsules, suitable for microencapsulation of hydrophobic materials with a wide range of polarities.


Fast Calculation of Computer Generated Holograms for 3D Photostimulation through Compressive-Sensing Gerchberg-Saxton Algorithm.
The use of spatial light modulators to project computer generated holograms is a common strategy for optogenetic stimulation of multiple structures of interest within a three-dimensional volume. A common requirement when addressing multiple targets sparsely distributed in three dimensions is the generation of a points cloud, focusing excitation light in multiple diffraction-limited locations throughout the sample. Calculation of this type of holograms is most commonly performed with either the high-speed, low-performance random superposition algorithm, or the low-speed, high performance Gerchberg-Saxton algorithm. This paper presents a variation of the Gerchberg-Saxton algorithm that, by only performing iterations on a subset of the data, according to compressive sensing principles, is rendered significantly faster while maintaining high quality outputs. The algorithm is presented in high-efficiency and high-uniformity variants. All source code for the method implementation is available as Supplementary Materials and as open-source software. The method was tested computationally against existing algorithms, and the results were confirmed experimentally on a custom setup for in-vivo multiphoton optogenetics. The results clearly show that the proposed method can achieve computational speed performances close to the random superposition algorithm, while retaining the high performance of the Gerchberg-Saxton algorithm, with a minimal hologram quality loss.


Function of UDP-glucuronosyltransferase 2B17 (UGT2B17) is involved in endometrial cancer.
Endometrial cancer (EC) is a steroid hormone-dependent cancer. Uridine 5'-diphospho-glucuronosyltransferase enzymes conjugate and detoxify endogenous and exogenous steroid hormones and environmental carcinogens. Among these enzymes, the function of UGT2B17 is unknown except for glucuronidation. The messenger RNA expression of UGT2B17 and myeloid cell leukemia-1 (Mcl-1) was significantly increased in EC tissues compared with matched normal endometrial tissues. Therefore, we focused on the function of UGT2B17 in EC. A total of nine patients with confirmed EC were enrolled in this study to investigate the expression of UGT2B17 and target genes. EC cell lines were used for functional tests including cell growth, invasion, apoptosis and cell cycle analyses. To find the target genes of UGT2B17, we performed microarray analysis to see which genes were upregulated or downregulated by UGT2B17-transfected cells. Functional analysis showed decreased numbers of viable cells and increased numbers of apoptotic cells in si-UGT2B17-transfected Ishikawa cells. Among microarray target genes, Mcl-1 was significantly downregulated in si-UGT2B17-transfected cells. We also found upregulation of Puma protein, a target of Mcl-1, in si-UGT2B17-transfected cells. This is the first report to show that UGT2B17 and Mcl-1 expression are upregulated in EC tissues and that UGT2B17 depletion induces inhibition of cell growth and apoptosis in EC cells through Mcl-1 downregulation.


[Interferon-inducing action of polysaccharide-containing biopolymers from ginseng root and cell culture].
Induction of interferon (IF) and tumor necrosis factor (TNF) under the action of two polysaccharide preparations of ginseng i.e. panaxan-1 (from ginseng root) and panaxan-2 (from ginseng cell culture) was studied. Both the preparations induced production of TNF and IF in human leukocytes. By its properties and the typing results the induced IF proved to be gamma-IF. The preparation from the ginseng cell culture in the doses used had a higher IF inducing activity which could be explained by the difference in the polysaccharide composition of the preparations.


The psychosis treatment gap and its consequences in rural Ethiopia.
The "treatment gap" (TG) for mental disorders, widely advocated by the WHO in low-and middle-income countries, is an important indicator of the extent to which a health system fails to meet the care needs of people with mental disorder at the population level. While there is limited research on the TG in these countries, there is even a greater paucity of studies looking at TG beyond a unidimensional understanding. This study explores several dimensions of the TG construct for people with psychosis in Sodo, a rural district in Ethiopia, and its implications for building a more holistic capacity for mental health services. The study was a cross-sectional survey of 300 adult participants with psychosis identified through community-based case detection and confirmed through subsequent structured clinical evaluations. The Butajira Treatment Gap Questionnaire (TGQ), a new customised tool with 83 items developed by the Ethiopia research team, was administered to evaluate several TG dimensions (access, adequacy and effectiveness of treatment, and impact/consequence of the treatment gap) across a range of provider types corresponding with the WHO pyramid service framework. Lifetime and current access gap for biomedical care were 41.8 and 59.9% respectively while the corresponding figures for faith and traditional healing (FTH) were 15.1 and 45.2%. Of those who had received biomedical care for their current episode, 71.7% did not receive minimally adequate care. Support from the community and non-governmental organisations (NGOs) were negligible. Those with education (Adj. OR: 2.1; 95% CI: 1.2, 3.8) and history of use of FTH (Adj. OR: 3.2; 95% CI: 1.9-5.4) were more likely to use biomedical care. Inadequate biomedical care was associated with increased lifetime risk of adverse experiences, such as history of restraint, homelessness, accidents and assaults. This is the first study of its kind. Viewing TG not as a unidimensional, but as a complex, multi-dimensional construct, offers a more realistic and holistic understanding of health beliefs, help-seeking behaviors, and need for care. The reconceptualized multidimensional TG construct could assist mental health services capacity building advocacy and policy efforts and allow community and NGOs play a larger role in supporting mental healthcare.


The decision sampling framework: a methodological approach to investigate evidence use in policy and programmatic innovation.
Calls have been made for greater application of the decision sciences to investigate and improve use of research evidence in mental health policy and practice. This article proposes a novel method, "decision sampling," to improve the study of decision-making and research evidence use in policy and programmatic innovation. An illustrative case study applies the decision sampling framework to investigate the decisions made by mid-level administrators when developing system-wide interventions to identify and treat the trauma of children entering foster care. Decision sampling grounds qualitative inquiry in decision analysis to elicit information about the decision-making process. Our case study engaged mid-level managers in public sector agencies (n = 32) from 12 states, anchoring responses on a recent index decision regarding universal trauma screening for children entering foster care. Qualitative semi-structured interviews inquired on questions aligned with key components of decision analysis, systematically collecting information on the index decisions, choices considered, information synthesized, expertise accessed, and ultimately the values expressed when selecting among available alternatives. Findings resulted in identification of a case-specific decision set, gaps in available evidence across the decision set, and an understanding of the values that guided decision-making. Specifically, respondents described 14 inter-related decision points summarized in five domains for adoption of universal trauma screening protocols, including (1) reach of the screening protocol, (2) content of the screening tool, (3) threshold for referral, (4) resources for screening startup and sustainment, and (5) system capacity to respond to identified needs. Respondents engaged a continuum of information that ranged from anecdote to research evidence, synthesizing multiple types of knowledge with their expertise. Policy, clinical, and delivery system experts were consulted to help address gaps in available information, prioritize specific information, and assess "fit to context." The role of values was revealed as participants evaluated potential trade-offs and selected among policy alternatives. The decision sampling framework is a novel methodological approach to investigate the decision-making process and ultimately aims to inform the development of future dissemination and implementation strategies by identifying the evidence gaps and values expressed by the decision-makers, themselves.


Severity of autism is related to children's language processing.
Problems in language processing have been associated with autism spectrum disorder (ASD), with some research attributing the problems to overall language skills rather than a diagnosis of ASD. Lexical access was assessed in a looking-while-listening task in three groups of 5- to 7-year-old children; two had high-functioning ASD (HFA), an ASD severe (ASD-S) group (n = 16) and an ASD moderate (ASD-M) group (n = 21). The third group were typically developing (TD) (n = 48). Participants heard sentences of the form "Where's the x?" and their eye movements to targets (e.g., train), phonological competitors (e.g., tree), and distractors were recorded. Proportions of looking time at target were analyzed within 200 ms intervals. Significant group differences were found between the ASD-S and TD groups only, at time intervals 1000-1200 and 1200-1400 ms postonset. The TD group was more likely to be fixated on target. These differences were maintained after adjusting for language, verbal and nonverbal IQ, and attention scores. An analysis using parent report of autistic-like behaviors showed higher scores to be associated with lower proportions of looking time at target, regardless of group. Further analysis showed fixation for the TD group to be significantly faster than for the ASD-S. In addition, incremental processing was found for all groups. The study findings suggest that severity of autistic behaviors will impact significantly on children's language processing in real life situations when exposed to syntactically complex material. They also show the value of using online methods for understanding how young children with ASD process language.


Regenerating the nephron with human pluripotent stem cells.
Nephrogenesis in humans is limited to the period of embryonic kidney development in utero, with no new nephrons formed after birth. Although the kidneys possess the capacity to self-repair segments of the nephron, nephron loss from acute or chronic kidney injury is irreversible and results in impaired function. Human pluripotent stem cells (PSCs), including embryonic stem cells and induced pluripotent stem cells, are an attractive source of cells to regenerate nephron progenitor cells (NPCs) and ultimately functional kidney tissue. NPCs are found exclusively during the period of embryonic development, but their nephron-forming capacity makes them an ideal cell population to regenerate with PSCs. Significant progress has been made in the effort to direct the differentiation of human PSCs into NPCs. Differentiation protocols designed to recapitulate the complex process of kidney organogenesis in vitro can generate cells that express characteristic NPC markers and these cells can assemble into three-dimensional nephron-like structures. Additional studies are required to evaluate the functionality of these putative kidney cells and to test their ability to integrate into three-dimensional organized kidney tissue structures, either spontaneously or facilitated by bioengineered structures or scaffolds with appropriate matrix materials. The successful recreation of human nephrons from PSCs would offer a novel therapeutic approach to treating patients with kidney disease.


Documentation of simulated shoulder dystocia: accurate and complete?
To investigate completeness and accuracy of record keeping by comparison of documentation and actual events, recorded on video and through a force-monitoring device, during simulated shoulder dystocia. An observational study. Six maternity units in South West of England. Seventy-one midwives and 39 doctors. Doctors and midwives documented their management of a shoulder dystocia simulation on paper used in their hospital (simple notepaper or preformatted form). Documentation was compared with video recording of each simulation and an electronic record of force applied during delivery. Documentation of head-to-body delivery time (and comparison with actual delivery time). Documentation of force (and comparison with actual applied force). A total of 110 participants documented their actions, 70.9% used a preformatted sheet, 29.1% used hospital notepaper. Fifty-six percent documented head-to-body delivery interval (HBDI) with 56% overestimating the time by more than 1 minute. Force used during the simulation was documented by 70.9%, with no relationship between the subjective description of force applied and the maximum recorded force. The anterior shoulder was documented by 78.2% and correctly identified in 80%. Documentation of force was more likely if a preformatted sheet was used (88 versus 53%, P = 0.016). Documentation of the laterality of the anterior shoulder was tended to be more accurate with plain hospital notepaper (93 versus 78%, P = 0.3526). Manoeuvres performed were well documented. HBDI and force applied were not documented accurately in the majority of simulated deliveries. Use of a preformatted sheet appeared to improve completeness, but not accuracy, of documentation.


Mechanisms, impact and prevention of pathological bone regeneration in spondyloarthritis.
To discuss different aspects of new bone formation in patients with spondyloarthritis based on emerging data from clinical trials, prospective cohort studies and translational laboratory investigations. New bone formation potentially leading to ankylosis of the spine and sacroiliac joints remains an important concern for patients with axial spondyloarthritis. New therapeutic strategies, in particular targeting of interleukin-17, have emerged in addition to the antitumor necrosis factor drugs, but we still fail to fully understand the mechanisms of structural disease progression. A new paradigm is developing in which sustained and effective suppression of inflammation likely inhibits this structural disease progression. Biomechanical factors, in particular changes in bone microarchitecture in the vertebrae, and the need for core stability could provide a new framework to understand the relationship between bone remodeling and inflammation and to develop long-term strategies. New bone formation leading to ankylosis remains a hallmark of axial spondyloarthritis and should be further investigated. The clinical data that progressively become available support the concept that effective and sustained therapy will be beneficial for the patients not only in short-term, but also in long-term outcomes.


Liposome drugs' loading efficiency: a working model based on loading conditions and drug's physicochemical properties.
Remote loading of liposomes by transmembrane gradients is one of the best approaches for achieving the high enough drug level per liposome required for the liposomal drug to be therapeutically efficacious. This breakthrough, which enabled the approval and clinical use of nanoliposomal drugs such as Doxil, has not been paralleled by an in-depth understanding that allows predicting loading efficiency of drugs. Here we describe how applying data-mining algorithms on a data bank based on Barenholz's laboratory's 15 years of liposome research experience on remote loading of 9 different drugs enabled us to build a model that relates drug physicochemical properties and loading conditions to loading efficiency. This model enables choosing candidate molecules for remote loading and optimizing loading conditions according to logical considerations. The model should also help in designing pro-drugs suitable for remote loading. Our approach is expected to improve and accelerate development of liposomal formulations for clinical applications.


Pharmacogenetic analysis of human steroid 5 alpha reductase type II: comparison of finasteride and dutasteride.
Human steroid 5 alpha-reductase type II is a prostate-specific, membrane-associated enzyme that catalyzes the conversion of testosterone to dihydrotestosterone, the most potent androgen in the prostate gland. Genetic variants of this enzyme have been associated with both the development and the progression of prostate cancer. Both finasteride and dutasteride are competitive inhibitors of the type II steroid 5 alpha-reductase that have been effectively used for the treatment of benign prostatic hyperplasia. Finasteride has also been successfully utilized for prostate cancer chemoprevention. We here investigate 5 alpha-reductase inhibition assays in vitro to measure the effect of incubation time on the apparent inhibition constant (Ki) for both constitutional and somatic (prostate cancer) enzyme variants. Our systematic pharmacogenetic analysis shows that both finasteride and dutasteride are slow, time-dependent inhibitors of steroid 5 alpha-reductase type II, and that the inhibition kinetics depend on the 5 alpha-reductase genotype. We also show that, overall, dutasteride is a more efficient steroid 5 alpha-reductase inhibitor than finasteride. Based on our data, we are able to map areas of the enzyme that are responsible for this time-dependent inhibition for either (or both) enzyme inhibitor(s). This comprehensive pharmacogenetic analysis of steroid 5 alpha-reductase variants unveiled significant pharmacogenetic variation for both finasteride and dutasteride and thus should be taken into account when designing protocols for treatment and/or chemoprevention of prostatic diseases with either one of these 5 alpha-reductase inhibitors since there is considerable pharmacogenetic variation for both drugs.


Illustrating idiographic methods for translation research: moderation effects, natural clinical experiments, and complex treatment-by-subgroup interactions.
A critical juncture in translation research involves the preliminary studies of intervention tools, provider training programs, policies, and other mechanisms used to leverage knowledge garnered at one translation stage into another stage. Potentially useful for such studies are rigorous techniques for conducting within-subject clinical trials, which have advanced incrementally over the last decade. However, these methods have largely not been utilized within prevention or translation contexts. The purpose of this manuscript is to demonstrate the flexibility, wide applicability, and rigor of idiographic clinical trials for preliminary testing of intervention mechanisms. Specifically demonstrated are novel uses of state-space modeling for testing intervention mechanisms of short-term outcomes, identifying heterogeneity in and moderation of within-person treatment mechanisms, a horizontal line plot to refine sampling design during the course of a clinic-based experimental study, and the need to test a treatment's efficacy as treatment is administered along with (e.g., traditional 12-month outcomes).