Adaptation of teleosts to very high salinity.
A number of species of euryhaline teleosts have the remarkable ability to adapt and survive in environments of extreme salinity, up to two or even three times the osmolality of seawater. This review looks at some of the literature describing the adaptive changes that occur, primarily with intestinal water absorption and with the properties of the gill epithelium. While there is much that is still not completely understood, recent work has begun to look at these adaptations at the cellular and molecular level. As with seawater osmoregulation, fish adapting to hypersaline conditions generally increase drinking rates and water absorption across the intestine by solute-linked transport. This process requires increased activity, expression levels and possibly changes in subunit isoforms of Na(+)/K(+)-ATPase pumps, along with increases in other components of the NaCl absorptive pathway. Additionally, recent studies have demonstrated the importance of luminal anion exchange activity and of luminal alkalinization, which can support absorption against increasing osmotic gradients by promoting divalent ion precipitation (CaCO(3)) and by conversion of HCO(3)(-) ions to CO(2). The steepness of the lumen to blood osmotic gradient vis-a-vis the NaCl absorptive capacity, along with the accumulation of non-absorbed Mg(++) and SO(4)(--), likely become the limiting factors for survival in hypersaline conditions. Of interest is the observation of predicted hyperosmotic fluid absorption by the intestine in several species. Adaptive changes in the gill epithelium are also critical in this process, allowing for secretion of absorbed NaCl from the extracellular fluids. Most notably there are important changes in the numbers and size of mitochondrion-rich (MR) cells, the sites of active secretion of Cl(-), which ultimately drives the overall process of NaCl secretion. Balance studies of intake and output clearly indicate that a decrease in the osmotic permeability of the gill epithelium must also occur. The molecular correlates of this effect are not known, although decreased expression of one or more aquaporins seems to be a likely possibility. Finally, the regulatory changes seen with hypersaline adaptation may provide important new insights into epithelial function, including the role of organized transport assemblies ("metabolons") and changes in the expression of tight junction proteins such as claudins or occludins, which may modulate electrolyte permeabilities.


Yoga reduces perceived stress and exhaustion levels in healthy elderly individuals.
This study investigated whether a 7-week yoga intervention could improve physical function, perceived stress, and mental/emotional wellness in elderly participants. 8 participants (66.5 ± 0.3 years) attended 2 60-min Hatha yoga sessions/week for 7 weeks, and performed pre- and post-intervention assessments. Balance was assessed using a 5-test battery. Flexibility was measured by sit-and-reach and shoulder flexibility tests. Functional mobility tests included 8-ft up-and-go, 5 chair stands, and 4-m walk. Participants completed SF-12, exhaustion level, and Perceived Stress Scale (PSS) questionnaires. SF-12 Mental Component Summary scores, exhaustion levels, and PSS scores improved post-intervention. No differences were found for physical function measures. Yoga participation can improve mental/emotional wellness, exhaustion levels, and stress levels in elderly individuals, even without measurable improvements in physical function. Clinicians and health practitioners who work with the elderly should consider yoga as a potential therapeutic modality for improving important aspects of quality of life in this population.


Novel 3-arylethynyl-substituted thieno[3,4-b]pyrazine derivatives as human transglutaminase 2 inhibitors.
In the process of optimization, we developed a novel core skeleton of thieno[3,4-b]pyrazine via GK-13. The derivatives synthesized were shown to inhibit TGase 2 activity in cancer cells. Some of the hit compounds such as the arylethynyl group-coupled thieno[3,4-b]pyrazine derivatives were shown to exhibit promising activity for use as potential therapeutic small-molecules in renal cancer by inhibiting TGase 2 activity.


Dermatopathic lymphadenitis associated with human papilloma virus infection and verruca vulgaris.
Here we present a pediatric case of human papilloma virus associated with dermatopathic lymphadenitis (DL). A 5-year-old boy presented to the pediatric oncology clinic with swelling of the neck and warts on his lower jaw. His blood chemistry and complete blood count were normal, as was chest x-ray. HIV, EBV, CMV, and parvovirus serologies were negative. The patient was investigated for malignancy and lymphoma but no association was found. Histopathologic examination of the lymph node and the lesion revealed DL and verruca vulgaris, respectively. DL represents a benign form of reactive lymph node hyperplasia and described in patients with HIV and EBV infections. It is a rare entity described in patients with human papilloma virus infection. To our knowledge, this is the first report of DL in a patient with human papilloma virus infection.


A pilot study on voucher-based incentives to promote abstinence from cigarette smoking during pregnancy and postpartum.
We report results from a pilot study examining the use of vouchers redeemable for retail items as incentives for smoking cessation during pregnancy and postpartum. Of 100 study-eligible women who were still smoking upon entering prenatal care, 58 were recruited from university-based and community obstetric practices to participate in a smoking cessation study. Participants were assigned to either contingent or noncontingent voucher conditions. Vouchers were available during pregnancy and for 12 weeks postpartum. In the contingent condition, vouchers were earned for biochemically verified smoking abstinence. In the noncontingent condition, vouchers were earned independent of smoking status. Abstinence monitoring and associated voucher delivery was conducted daily during the initial 5 days of the cessation effort, gradually decreased to every other week antepartum, increased to once weekly during the initial 4 weeks postpartum, and then decreased again to every other week for the remaining 8 weeks of the postpartum intervention period. Contingent vouchers increased 7-day point-prevalence abstinence at the end-of-pregnancy (37% vs. 9%) and 12-week postpartum (33% vs. 0%) assessments. That effect was sustained through the 24-week postpartum assessment (27% vs. 0%), which was 12 weeks after discontinuation of the voucher program. Total mean voucher earnings across antepartum and postpartum were 397 US dollars (SD=414 US dollars) and 313 US dollars (SD=142 dollars) in the contingent and noncontingent conditions, respectively. The magnitude of these treatment effects exceed levels typically observed with pregnant and recently postpartum smokers, and the maintenance of effects through 24 weeks postpartum extends the duration beyond those reported previously.


Decoupled pelvis rotation in sitting: a passive motion technique that regulates buttock load associated with pressure ulcer development.
Wheelchair-users who cannot reposition themselves often suffer from pressure ulcers which are places of tissue breakdown in the buttock region under the sacrum and ischial tuberosities. Periodic pressure relief is needed to recover the buttock tissue from continuous deformation and impairment of tissue perfusion. Because pelvis alignment directly affects body posture and buttock load, a passive motion technique was developed that adjusts pelvis orientation independent from the trunk and seat support. This study investigates the effects of the so-called decoupled pelvis rotation (DPR) on the loads at the buttock-seat interface and evaluates whether this technique is applicable to regulate buttock load in sitting. Eighteen healthy male subjects participated in this study. Experiments were performed with a computer-aided adjustable simulator chair, instrumented with a concept of DPR. Measurements involved quasi-static actuated pelvis movements in the sagittal and frontal plane. Pelvis orientation, buttock interface pressures, seat reaction forces and centre of pressure were simultaneously measured. As a result of the induced passive pelvis movements, the pelvis rotated 19+/-2 degrees and 9+/-2 degrees in the sagittal and frontal plane, respectively. Significant relations were found between pelvis rotation and most quantities of buttock load. Findings suggest that DPR is an effective technique to regulate buttock load in able-bodied individuals. For clinical application, this technique is still to be evaluated on individuals who cannot functionally reposition themselves. Impairments to the neuromuscular function influence postural response from chair adjustments which makes clinical investigation necessary.


Fabrication and Characterization of a Capacitive Photodetector Comprising a ZnS/Cu Particle/Poly(vinyl butyral) Composite.
Most photodetectors developed to date essentially measure photocurrents induced by the generation and separation of electron-hole pairs in semiconductors during irradiation. Although the above light detection method is well established, highly sensitive, and applicable to a broad range of semiconductor materials, it requires the presence of a stable and direct contact between the semiconductor and the electrode for accurate photocurrent measurements. In turn, this prerequisite necessitates the use of various costly processes for device fabrication (e.g., photolithography and vacuum deposition of semiconductors/metals) and complicates the development of flexible devices. Herein, inspired by the fact that the dielectric properties of certain materials can be changed by light irradiation, we dispersed ZnS/Cu semiconducting particles in poly(vinyl butyral) to prepare a free-standing composite film and formed two layers of Ag nanowire electrodes on both sides of the cured composite to fabricate a photodetector of a completely new type. The developed device exhibited a capacitance very sensitive to irradiation with light of a specific wavelength and additionally featured the advantages of simple structure/operation mechanism, mechanical flexibility, and transparency, not showing any signs of performance deterioration even after severe damage.


The phosphorylation of CapZ-interacting protein (CapZIP) by stress-activated protein kinases triggers its dissociation from CapZ.
A protein expressed in immune cells and muscle was detected in muscle extracts as a substrate for several SAPKs (stress-activated protein kinases). It interacted specifically with the F-actin capping protein CapZ in splenocytes, and was therefore termed 'CapZIP' (CapZ-interacting protein). Human CapZIP was phosphorylated at Ser-179 and Ser-244 by MAPKAP-K2 (mitogen-activated protein kinase-activated protein kinase 2) or MAPKAP-K3 in vitro. Anisomycin induced the phosphorylation of CapZIP at Ser-179 in Jurkat cells, which was prevented by SB 203580, consistent with phosphorylation by MAPKAP-K2 and/or MAPKAP-K3. However, osmotic shock-induced phosphorylation of Ser-179 was unaffected by SB 203580. These and other results suggest that CapZIP is phosphorylated at Ser-179 in cells by MAPKAP-K2/MAPKAP-K3, and at least one other protein kinase. Stress-activated MAP kinase family members phosphorylated human CapZIP at many sites, including Ser-68, Ser-83, Ser-108 and Ser-216. Ser-108 became phosphorylated when Jurkat cells were exposed to osmotic shock, which was unaffected by SB 203580 and/or PD 184352, or in splenocytes from mice that do not express either SAPK3/p38gamma or SAPK4/p38delta. Our results suggest that CapZIP may be phosphorylated by JNK (c-Jun N-terminal kinase), which phosphorylates CapZIP to >5 mol/mol within minutes in vitro. Osmotic shock or anisomycin triggered the dissociation of CapZIP from CapZ in Jurkat cells, suggesting that phosphorylation of CapZIP may regulate the ability of CapZ to remodel actin filament assembly in vivo.


Thermal characterization of carbon nanotube foam using MEMS microhotplates and thermographic analysis.
Thermal material properties play a fundamental role in the thermal management of microelectronic systems. The porous nature of carbon nanotube (CNT) arrays results in a very high surface area to volume ratio, which makes the material attractive for surface driven heat transfer mechanisms. Here, we report on the heat transfer performance of lithographically defined micropins made of carbon nanotube (CNT) nanofoam, directly grown on microhotplates (MHPs). The MHP is used as an in situ characterization platform with controllable hot-spot and integrated temperature sensor. Under natural convection, and equivalent power supplied, we measured a significant reduction in hot-spot temperature when augmenting the MHP surface with CNT micropins. In particular, a strong enhancement of convective and radiative heat transfer towards the surrounding environment is recorded, due to the high aspect ratio and the foam-like morphology of the patterned CNTs. By combining electrical characterizations with high-resolution thermographic microscopy analysis, we quantified the heat losses induced by the integrated CNT nanofoams and we found a unique temperature dependency of the equivalent convective heat transfer coefficient, Hc. The obtained results with the proposed non-destructive characterization method demonstrate that significant improvements can be achieved in microelectronic thermal management and hierarchical structured porous material characterization.


Effects of gamma-glutamyl hydrolase on folyl and antifolylpolyglutamates in cultured H35 hepatoma cells.
A subline of H35 hepatoma cells (H35D cells) that have been made resistant to 5,10-dideazatetrahydrofolate exhibits an increase in gamma-glutamyl hydrolase (GH) activity. GH is a lysosomal enzyme in H35 and H35D cells on the basis of comparison of the distribution of enzyme activity with other known lysosomal enzymes. The hydrolysis rate of methotrexate polyglutamate with isolated, intact lysosomes is 4-5-fold greater in H35D cells than in H35 cells. GH activity in isolated lysosomes is in part dependent on the presence of a reducing agent such as mercaptoethanol. Permeabilization of lysosomal preparations from both cell types by Triton X-100 causes a 10-fold enhancement in GH activity. The result of the enhanced activity of GH in H35D cells is a marked reduction in antifolylpolyglutamate concentration, with the parent antifolate being the predominant intracellular species found under all conditions tested. Unlike antifolates, the total intracellular folate concentration is nearly identical in both cells under standard culture conditions up to 10 microns folic acid. However, the chain length of folylpolyglutamates consists of predominantly triglutamates and tetraglutamates in H35D cells with increased GH, whereas it consists of pentaglutamates and hexaglutamates in the parental cells. At 50 and 100 microns folic acid, the folate accumulation in H35D cells is less than half that of H35 cells, and the predominant polyglutamate species in the H35D cells are the diglutamates through the tetraglutamates. The results demonstrate that the two H35 cell lines having equal folylpolyglutamate synthetase but that one with enhanced lysosomal GH activity exhibits a marked reduction in the amount and gamma-glutamyl chain length of folylpolyglutamates and antifolylpolyglutamates.


Geographical variation and population structure in the White-rumped Sandpiper Calidris fuscicollis as shown by morphology, mitochondrial DNA and carbon isotope ratios.
We studied the population structure of a high arctic breeding wader bird species, the White-rumped Sandpiper Calidris fuscicollis. Breeding adults, chicks and juveniles were sampled at seven localities throughout the species' breeding range in arctic Canada in 1999. The mitochondrial control region was analysed by DNA sequencing, feathers were analysed for carbon isotope ratios (C13/C12) by isotope ratio mass spectrometry, and morphological measurements were analysed using principal component analyses, taking the effect of sex into account (identified by molecular genetic methods). In general, our results support the notion that the White-rumped Sandpiper is a monotypic species with no subspecies, and most of the morphological and genetic variation occurs within sites. Nevertheless, some differences between sites were found. Birds from the two northernmost sites (Ellesmere and Devon Islands) had relatively longer bill and wing and shorter tarsus than birds sampled further south, possibly reflecting genetic differences between populations. The carbon isotope ratios were higher at the easternmost site (Baffin Island), revealing differences in the isotope content of the food. The mtDNA sequences showed no significant differentiation between sites and no pattern of isolation-by-distance was found. Based on the mtDNA variation, the species was estimated to have a long-term effective population size of approximately 9,000 females. The species shows no clear evidence of any population expansion or decline. Our results indicate that carbon isotope ratios, and possibly also certain mtDNA haplotypes, may be useful as tools for identifying the breeding origin of White-rumped Sandpipers on migration and wintering sites.


The best COVID-19 predictor is recent smell loss: a cross-sectional study.
COVID-19 has heterogeneous manifestations, though one of the most common symptoms is a sudden loss of smell (anosmia or hyposmia). We investigated whether olfactory loss is a reliable predictor of COVID-19. This preregistered, cross-sectional study used a crowdsourced questionnaire in 23 languages to assess symptoms in individuals self-reporting recent respiratory illness. We quantified changes in chemosensory abilities during the course of the respiratory illness using 0-100 visual analog scales (VAS) for participants reporting a positive (C19+; n=4148) or negative (C19-; n=546) COVID-19 laboratory test outcome. Logistic regression models identified singular and cumulative predictors of COVID-19 status and post-COVID-19 olfactory recovery. Both C19+ and C19- groups exhibited smell loss, but it was significantly larger in C19+ participants (mean±SD, C19+: -82.5±27.2 points; C19-: -59.8±37.7). Smell loss during illness was the best predictor of COVID-19 in both single and cumulative feature models (ROC AUC=0.72), with additional features providing no significant model improvement. VAS ratings of smell loss were more predictive than binary chemosensory yes/no-questions or other cardinal symptoms, such as fever or cough. Olfactory recovery within 40 days was reported for ~50% of participants and was best predicted by time since illness onset. As smell loss is the best predictor of COVID-19, we developed the ODoR-19 tool, a 0-10 scale to screen for recent olfactory loss. Numeric ratings ≤2 indicate high odds of symptomatic COVID-19 (10<OR<4), especially when viral lab tests are impractical or unavailable.


Retrospective analysis of posttransplantation diabetes mellitus in black renal allograft recipients.
To study the incidence, outcome, and possible etiopathogenic factors involved in posttransplantation diabetes mellitus in cyclosporine-treated black renal allograft recipients. One hundred thirty-eight nondiabetic black renal transplant recipients whose grafts survived greater than 1 yr were studied retrospectively. Twenty-eight (20.3%) patients developed posttransplantation diabetes mellitus, 46 and 75% were diagnosed by 6- and 12-mo posttransplantation, respectively, and 46% were insulin dependent. Diabetes was more frequently encountered in older recipients and recipients of cadaveric kidneys but was independent of sex, number of transplants, incidence of acute rejection, percentage of body weight gain, steroid or cyclosporine dose, and use of beta-blockers and/or diuretics. Renal function was similar in the diabetic group compared with the control group. Actuarial 5-yr graft survival was 82% in the diabetic cohort compared with 78% in the control group, with chronic rejection accounting for all graft losses within the diabetic group. Twenty percent of black cyclosporine-treated renal allograft recipients developed diabetes mellitus in the posttransplantation period. However, its presence did not appear to influence intermediate-term graft or patient survival.


Collaborative active learning: bioimpedance and anthropometry in higher education.
Research in the health sciences devotes much attention to overweight and obesity and, consequently, to body composition. In recent years, traditional body measures have been questioned as efficient variables in health sciences due to the fact that they cannot give information about body fat mass. Our aim is to teach how to analyze body composition through anthropometry and bioelectrical impedance analysis to our "Physiology of Vegetative and Reproductive Functions" students, who are studying for their degree in Biology. We proposed project-oriented-learning to promote collaborative interactions among students. Fifty-two students voluntarily formed five groups; they worked with the concepts of basal metabolic rate and body composition from a theoretical point of view and later transformed these concepts into a practical perspective by preparing a manuscript in groups with objectives proposed by our teaching team. In this research, we show a collaborative educational scenario for university students in which students are tutored from a constructivist perspective to promote social interactions, resulting in new knowledge acquisition.


Diversity in global maize germplasm: characterization and utilization.
Maize (Zea mays L.) is not only of worldwide importance as a food, feed and as a source of diverse industrially important products, but is also a model genetic organism with immense genetic diversity. Although it was first domesticated in Mexico, maize landraces are widely found across the continents. Several studies in Mexico and other countries highlighted the genetic variability in the maize germplasm. Applications of molecular markers, particularly in the last two decades, have led to new insights into the patterns of genetic diversity in maize globally, including landraces as well as wild relatives (especially teosintes) in Latin America, helping in tracking the migration routes of maize from the centers of origin, and understanding the fate of genetic diversity during maize domestication. The genome sequencing of B73 (a highly popular US Corn Belt inbred) and Palomero (a popcorn landrace in Mexico) in the recent years are important landmarks in maize research, with significant implications to our understanding of the maize genome organization and evolution. Next-generation sequencing and high-throughput genotyping platforms promise to further revolutionize our understanding of genetic diversity and for designing strategies to utilize the genomic information for maize improvement. However, the major limiting factor to exploit the genetic diversity in crops like maize is no longer genotyping, but high-throughput and precision phenotyping. There is an urgent need to establish a global phenotyping network for comprehensive and efficient characterization of maize germplasm for an array of target traits, particularly for biotic and abiotic stress tolerance and nutritional quality. 'Seeds of Discovery' (SeeD), a novel initiative by CIMMYT with financial support from the Mexican Government for generating international public goods, has initiated intensive exploration of phenotypic and molecular diversity of maize germplasm conserved in the CIMMYT Gene Bank; this is expected to aid in effective identification and use of novel alleles and haplotypes for maize improvement. Multi-institutional efforts are required at the global level to systematically explore the maize germplasm to diversify the genetic base of elite breeding materials, create novel varieties and counter the effects of global climate changes.


T2 mapping of the heart with a double-inversion radial fast spin-echo method with indirect echo compensation.
The abnormal signal intensity in cardiac T2-weighted images is associated with various pathologies including myocardial edema. However, the assessment of pathologies based on signal intensity is affected by the acquisition parameters and the sensitivities of the receiver coils. T2 mapping has been proposed to overcome limitations of T2-weighted imaging, but most methods are limited in spatial and/or temporal resolution. Here we present and evaluate a double inversion recovery radial fast spin-echo (DIR-RADFSE) technique that yields data with high spatiotemporal resolution for cardiac T2 mapping. DIR-RADFSE data were collected at 1.5 T on phantoms and subjects with echo train length (ETL) = 16, receiver bandwidth (BW) = ±32 kHz, TR = 1RR, matrix size = 256 × 256. Since only 16 views per echo time (TE) are collected, two algorithms designed to reconstruct highly undersampled radial data were used to generate images for 16 time points: the Echo-Sharing (ES) and the CUrve Reconstruction via pca-based Linearization with Indirect Echo compensation (CURLIE) algorithm. T2 maps were generated via least-squares fitting or the Slice-resolved Extended Phase Graph (SEPG) model fitting. The CURLIE-SEPG algorithm accounts for the effect of indirect echoes. The algorithms were compared based on reproducibility, using Bland-Altman analysis on data from 7 healthy volunteers, and T2 accuracy (against a single-echo spin-echo technique) using phantoms. Both reconstruction algorithms generated in vivo images with high spatiotemporal resolution and showed good reproducibility. Mean T2 difference between repeated measures and the coefficient of repeatability were 0.58 ms and 2.97 for ES and 0.09 ms and 4.85 for CURLIE-SEPG. In vivo T2 estimates from ES were higher than those from CURLIE-SEPG. In phantoms, CURLIE-SEPG yielded more accurate T2s compared to reference values (error was 7.5-13.9% for ES and 0.6-2.1% for CURLIE-SEPG), consistent with the fact that CURLIE-SEPG compensates for the effects of indirect echoes. The potential of T2 mapping with CURLIE-SEPG is demonstrated in two subjects with known heart disease. Elevated T2 values were observed in areas of suspected pathology. DIR-RADFSE yielded TE images with high spatiotemporal resolution. Two algorithms for generating T2 maps from highly undersampled data were evaluated in terms of accuracy and reproducibility. Results showed that CURLIE-SEPG yields T2 estimates that are reproducible and more accurate than ES.


Myocardial Bridging: A Case Presentation of Atypical Chest Pain Syndrome in a Young Woman.
BACKGROUND Myocardial bridging, although frequent, is often a forgotten cause of angina. It is a congenital anomaly in which the coronary artery tunnels through the myocardium with the overlying muscle, termed a myocardial bridge. The tunneled artery is prone to increased myocardial compression, mechanical load, endothelial damage, and vascular remodeling. During myocardial systole, the tunneled artery undergoes narrowing caused by myocardial compression, which leads to disruption of blood flow, thereby precipitating angina, arrhythmias, infarction, and sudden cardiac death. CASE REPORT Here, we present a case of a 29-year-old white woman who presented with atypical left-sided achy chest pain, occurring primarily at rest. Further evaluation showed mildly elevated troponins, with normal electrocardiogram, chest x-ray, and CTA chest. She subsequently underwent coronary angiography and was found to have myocardial bridging of her left anterior descending artery, with compression of up to 40% during systole. She was initially treated with diltiazem, but due to adverse effects was transitioned to metoprolol succinate, which she has tolerated well. CONCLUSIONS Myocardial bridging, although benign in nature, carries a vast array of complications requiring these patients to undergo prompt diagnosis and treatment. Vascular spasm, wall stress of the tunneled artery, and intensity of systolic constriction coupled with any delay in management can lead to ischemia, infarction, dysrhythmias, and death. Therefore, it is imperative that patients who have low clinical suspicion for atherosclerosis but who are presenting with anginal equivalents undergo coronary angiography to assess for myocardial bridging and receive immediate treatment.


Challenges and opportunities in augmentative and alternative communication: Research and technology development to enhance communication and participation for individuals with complex communication needs.
The field of augmentative and alternative communication (AAC) has witnessed significant changes since its inception. AAC services are now considered for a much greater number of individuals with complex communication needs and there are many more AAC options available as communication supports, including a proliferation of technologies. The scope and options for communication within society have increased substantially to include a wide array of digital and social media. Individuals with complex communication needs have increased expectations for participation and engagement across a full range of environments-education, employment, family, healthcare, and community living. Despite these advances, there remain critical challenges that must be addressed. This paper discusses key advances in the AAC field, delineates challenges, and discusses future directions to address these challenges, specifically as they relate to research and development to enhance AAC interventions and technologies for individuals with complex communication needs and their families.


Relieving occupational fatigue by consumption of a beverage containing γ-amino butyric acid.
To elucidate the effect of γ-aminobutyric acid (GABA) on both psychological and physical fatigue and on the performance advances for task solving, we assigned an arithmetic task for the Uchida-Kraepelin Psychodiagnostic Test (UKT) to 30 healthy Japanese subjects, 9 of whom were diagnosed as having chronic fatigue. The subjects were administered 250 mL of a test beverage containing GABA at the dose of 0, 25, and 50 mg before assigning task for the UKT. Psychological fatigue assessed by the Visual Analogue Scale (VAS) was significantly lower in the group administrated the beverage containing 50 mg GABA than in the control group (p<0.05). The results of the Profile of Mood States (POMS) also indicated that psychological fatigue was significantly reduced in the 50-mg-GABA group. The salivary secretion levels of chromogranin A and cortisol-markers of physical fatigue-in both 25-mg and 50-mg-GABA groups were significantly lower than those in the control group. The 50-mg-GABA group also showed higher score on UKT by solving the arithmetic task more accurately than the control group (p<0.01). The results suggest that intake of GABA-containing beverages, especially those containing 50 mg of GABA, may help reduce both psychological and physical fatigue and improve task-solving ability.


Biology and genome of a newly discovered sibling species of Caenorhabditis elegans.
A 'sibling' species of the model organism Caenorhabditis elegans has long been sought for use in comparative analyses that would enable deep evolutionary interpretations of biological phenomena. Here, we describe the first sibling species of C. elegans, C. inopinata n. sp., isolated from fig syconia in Okinawa, Japan. We investigate the morphology, developmental processes and behaviour of C. inopinata, which differ significantly from those of C. elegans. The 123-Mb C. inopinata genome was sequenced and assembled into six nuclear chromosomes, allowing delineation of Caenorhabditis genome evolution and revealing unique characteristics, such as highly expanded transposable elements that might have contributed to the genome evolution of C. inopinata. In addition, C. inopinata exhibits massive gene losses in chemoreceptor gene families, which could be correlated with its limited habitat area. We have developed genetic and molecular techniques for C. inopinata; thus C. inopinata provides an exciting new platform for comparative evolutionary studies.


Whole genome assembly of the armored loricariid catfish Ancistrus triradiatus highlights herbivory signatures.
The catfish Ancistrus triradiatus belongs to the species-rich family Loricariidae. Loricariids display remarkable traits such as herbivory, a benthic lifestyle, the absence of scales but the presence of dermal bony plates. They are exported as ornamental fish worldwide, with escaped fishes becoming a threat locally. Although genetic and phylogenetic studies are continuously increasing and developmental genetic investigations are underway, no genome assembly has been formally proposed for Loricariidae yet. We report a high-quality genome assembly of Ancistrus triradiatus using long and short reads, and a newly assembled transcriptome. The genome assembly is composed of 9530 scaffolds, including 85.6% of ray-finned fish BUSCOs, and 26,885 predicted protein-coding genes. The genomic GC content is higher than in other catfishes, reflecting the higher metabolism associated with herbivory. The examination of the SCPP gene family indicates that the genes presumably triggering scale loss when absent, are present in the scaleless A. triradiatus, questioning their explanatory role. The analysis of the opsin gene repertoire revealed that gene losses associated to the nocturnal lifestyle of catfishes were not entirely found in A. triradiatus, as the UV-sensitive opsin 5 is present. Finally, most gene family expansions were related to immunity except the gamma crystallin gene family which controls pupil shape and sub-aquatic vision. Thus, the genome of A. triradiatus reveals that fish herbivory may be related to the photic zone habitat, conditions metabolism, photoreception and visual functions. This genome is the first for the catfish suborder Loricarioidei and will serve as backbone for future genetic, developmental and conservation studies.


Water-actuated microcapsules fabricated by microfluidics.
We found a new water-actuated feature of poly(N-isopropylacrylamide) microgels and fabricated microcapsules with this feature based on microfluidic double emulsions. The microcapsules would release encapsulated actives by simple hydration, while forming biphasic hybrid microparticles by gradual dehydration. More complex microcapsules and hybrid microparticles could be produced by varying flow rates and inner oil types. These novel microcapsules could potentially be used for controllable storage or release of chemicals, fabrication of complex microparticles and applications in biochemical fields.


The effect of Mindfulness-Based Stress Reduction on wound healing: a preliminary study.
Psychological factors have been shown to influence the process of wound healing. This study examined the effect of Mindfulness-Based Stress Reduction (MBSR) on the speed of wound healing. The local production of pro-inflammatory cytokines and growth factors was studied as potential underlying mechanism. Forty-nine adults were randomly allocated to a waiting-list control group (n = 26) or an 8-week MBSR group (n = 23). Pre- and post-intervention/waiting period assessment for both groups consisted of questionnaires. Standardized skin wounds were induced on the forearm using a suction blister method. Primary outcomes were skin permeability and reduction in wound size monitored once a day at day 3, 4, 5, 6, 7, and 10 after injury. Secondary outcomes were cytokines and growth factors and were measured in wound exudates obtained at 3, 6, and 22 h after wounding. Although there was no overall condition effect on skin permeability or wound size, post hoc analyses indicated that larger increases in mindfulness were related to greater reductions in skin permeability 3 and 4 days after wound induction. In addition, MBSR was associated with lower levels of interleukin (IL)-8 and placental growth factor in the wound fluid 22 h after wound induction. These outcomes suggest that increasing mindfulness by MBSR might have beneficial effects on early stages of wound healing. Trial Registration NTR3652, http://www.trialregister.nl.


Cardiovascular risk among adults with chronic kidney disease, with or without prior myocardial infarction.
This study sought to determine whether chronic kidney disease (CKD) should be considered a coronary heart disease (CHD) risk equivalent for cholesterol treatment. It is unclear whether patients with CKD have a risk of CHD events or cardiovascular disease (CVD) mortality equivalent to patients with a prior myocardial infarction (MI). Using data from the ARIC (Atherosclerosis Risk in Communities) study, we categorized nondiabetic participants based on their average level of kidney function (estimated glomerular filtration rate > or =60 or 30 to 59 ml/min/1.73 m2, which defines stage 3 CKD) and on prior MI (yes or no). Rates and relative risks (RR) of CHD (MI or fatal CHD) events (n = 653) and CVD mortality (n = 209) that occurred over 10 years were compared across these populations. Among 12,243 middle-age participants, 271 had stage 3 CKD. After adjustment for age, gender, race, and center, CHD incidence and CVD mortality rates per 1,000 person-years by presence of CKD and MI were 4.1 and 1.0 in the presence of neither condition, 8.0 and 3.4 in CKD only, 18.8 and 7.0 in MI only, and 30.8 and 18.0 in CKD and MI. After further adjustment for CVD risk factors, RR of CHD and CVD mortality were statistically significantly lower in subjects with CKD and no prior MI (RR = 0.44 [95% confidence interval (CI) 0.28 to 0.72] for CHD and RR = 0.46 [95% CI 0.24 to 0.90] for CVD mortality) than for subjects with no CKD and a prior MI. Stage 3 CKD confers CHD risk that is lower and not equivalent to a prior MI in this middle-aged, general, nondiabetic population.


First Description of a Yersinia pseudotuberculosis Clonal Outbreak in France, Confirmed Using a New Core Genome Multilocus Sequence Typing Method.
Yersinia pseudotuberculosis is an enteric pathogen causing mild enteritis that can lead to mesenteric adenitis in children and septicemia in elderly patients. Most cases are sporadic, but outbreaks have already been described in different countries. We report for the first time a Y. pseudotuberculosis clonal outbreak in France, that occurred in 2020. An epidemiological investigation based on food queries pointed toward the consumption of tomatoes as the suspected source of infection. The Yersinia National Reference Laboratory (YNRL) developed a new cgMLST scheme with 1,921 genes specific to Y. pseudotuberculosis that identified the clustering of isolates associated with the outbreak and allowed to perform molecular typing in real time. In addition, this method allowed to retrospectively identify isolates belonging to this cluster from earlier in 2020. This method, which does not require specific bioinformatic skills, is now used systematically at the YNRL and proves to display an excellent discriminatory power and is available to the scientific community. IMPORTANCE We describe in here a novel core-genome MLST method that allowed to identify in real time, and for the first time in France, a Y. pseudotuberculosis clonal outbreak that took place during the summer 2020 in Corsica. Our method allows to support epidemiological and microbiological investigations to establish a link between patients infected with closely associated Y. pseudotuberculosis isolates, and to identify the potential source of infection. In addition, we made this method available for the scientific community.


Induction of acute inflammation in vivo by staphylococcal superantigens I: Leukocyte recruitment occurs independently of T lymphocytes and major histocompatibility complex Class II molecules.
Studies in our laboratory and others have recently shown that staphylococcal enterotoxin-derived superantigens stimulate proinflammatory cytokine gene expression in vitro. We have therefore investigated the ability of superantigens to induce leukocyte accumulation at extravascular sites in vivo using the subcutaneous air pouch model. Injection of staphylococcal enterotoxin A (SEA) induced a significant accumulation of leukocytes over basal levels in a time- and dose-dependent manner. It was also shown that superantigens are capable of inducing this response in mice depleted of CD4 T cells, as well as in severe combined immune-deficient and nude mice. These observations suggest that superantigens are capable of inducing leukocyte accumulation independently of the presence of T lymphocytes. Experiments were also conducted using mutant SEAs that have a reduced binding affinity for major histocompatibility complex (MHC) Class II molecules, as well as using MHC Class II-deficient mice. The results of these experiments indicated that MHC Class II molecules are not required for the observed effect of superantigens in vivo. Taken together, these results indicate, first, that bacterial superantigens promote inflammation in subcutaneous tissue in vivo and, second, the potential existence of a novel receptor for superantigens that mediates this subcutaneous inflammatory response.


A checklist that enhances the provision of education during insulin initiation simulation: a randomized controlled trial.
The study tested the hypothesis that doctors using an insulin information checklist during simulated insulin initiation would impart more information regarding insulin use. A total of 128 simulations were conducted. Doctors (n = 64) were recruited from practitioners recently completing internship (n = 19) and those established in primary care (n = 45). Both groups of doctors were strata randomized to control (n = 32) and intervention groups (n = 32), so that each group contained equal numbers. Doctors in each group experienced two identical simulations of insulin initiation with an intervening period of 10 min. Doctors in the intervention arm were introduced to an insulin initiation checklist, which they reviewed independently and utilized in the second simulation. Trained assessors captured the provision of education in 21 predefined educational areas. Differences in the change of the total education provided between the first and second simulations were assessed using linear regression. The difference in the mean change of education provided between the first and second simulations within the 21 educational areas for the control and treatment groups was 9.7 [95% confidence interval (CI): 8.8-11.1, P < 0.001] - an increase of 46.2%. The difference for the 15 areas relevant to pen use was 7.3 (95% CI: 6.2-8.4, P < 0.001) - an increase of 51.6%. The checklist resulted in doctors providing significantly more education applicable to syringe and insulin pen routes of insulin administration during simulations. Further research is needed on the checklist's impact on healthcare professionals and patient outcomes in the clinical context. (Clinical Trials Registry No: NCT02266303).


Multi-state outcome analysis of treatments (MOAT): application of a new approach to evaluate outcomes in longitudinal studies of bipolar disorder.
Survival analyzes are usually based on a single point in time predefined event. Dissatisfied with this approach to evaluating maintenance treatment outcomes, we developed the Multi-state Outcome Analysis of Treatments (MOAT) methodology using a combined database from two FDA registration studies of lamotrigine, lithium and placebo. MOAT partitions total survival time into clinically distinct periods operationally defined by cutpoints on rating scales. For bipolar disorder (BD), the clinical states are remission, subsyndromal and syndromal mania, mixed states or depression. MOAT results can be crossed with information about tolerability and functioning to yield an outcome system integrating efficacy and tolerability. As found in the original analysis, both drugs were associated with longer time in study compared with the placebo. MOAT supplements this by finding that both drugs increased the time remitted compared with placebo. However, a substantial amount of time in all three treatments was spent in subsyndromal depression. Time with manic symptoms was reduced with lithium, but not lamotrigine. Patients on placebo neither benefitted nor had adverse effects from the assignment but experienced more syndromal levels of symptoms and were terminated from the study sooner than either drug treated group. Lithium was associated with both benefit in time manic and worse tolerability compared with placebo. In summary, lamotrigine was associated with limited therapeutic benefit but not harm; lithium with both benefit and harm; and placebo with neither. MOAT describes not only quantity but also quality of time spent in longitudinal studies, providing a more clinically informative picture than Kaplan-Meier survival analysis.


Intrathecal ropivacaine in cesarean delivery.
The aim of the present study was to evaluate the optimum dose of ropivacaine by comparing three different dosing regimens of isobaric ropivacaine 1% (naropin 10 mg/ml, Astra Zeneca) administered intrathecally and to demonstrate the effects of anesthesia in pregnant women scheduled for cesarean section. Sixty ASA grade I-II patients were scheduled to undergo elective cesarean sections under spinal anesthesia. The patients were randomly assigned into three groups. Group 1 received 15 mg ropivacaine 1%, Group 2 received 20 mg ropivacaine 1%, and Group 3 received 25 mg ropivacaine 1%. Intraoperative hemodynamic variables were not significantly different between the three groups, and sensory block time, motor block time and time to reach maximal sensory block time, and motor block time were similar between the three groups. The time to two-segment regression of sensory block was longer in Group 3 compared to other groups, and the difference was statistically significant (p < 0.05). The motor block time was longer with higher doses of ropivacaine; however, the difference was not statistically significant. Ropivacaine administration produced rapid induction of anesthesia and satisfactory anesthesia level, ropivacaine 15 mg and 20 mg dosing regimens are satisfactory for spinal anesthesia.


Mixed and Multi-Methods Protocol to Evaluate Implementation Processes and Early Effects of the Pradhan Mantri Jan Arogya Yojana Scheme in Seven Indian States.
In September 2018, India launched Pradhan Mantri Jan Arogya Yojana (PM-JAY), a nationally implemented government-funded health insurance scheme to improve access to quality inpatient care, increase financial protection, and reduce unmet need for the most vulnerable population groups. This protocol describes the methodology adopted to evaluate implementation processes and early effects of PM-JAY in seven Indian states. The study adopts a mixed and multi-methods concurrent triangulation design including three components: 1. demand-side household study, including a structured survey and qualitative elements, to quantify and understand PM-JAY reach and its effect on insurance awareness, health service utilization, and financial protection; 2. supply-side hospital-based survey encompassing both quantitative and qualitative elements to assess the effect of PM-JAY on quality of service delivery and to explore healthcare providers' experiences with scheme implementation; and 3. process documentation to examine implementation processes in selected states transitioning from either no or prior health insurance to PM-JAY. Descriptive statistics and quasi-experimental methods will be used to analyze quantitative data, while thematic analysis will be used to analyze qualitative data. The study design presented represents the first effort to jointly evaluate implementation processes and early effects of the largest government-funded health insurance scheme ever launched in India.


All-time low period fertility in Finland: Demographic drivers, tempo effects, and cohort implications.
The ongoing period fertility decline in the Nordic countries is particularly strong in Finland, where the total fertility rate (TFR) reached an all-time low of 1.41 in 2018. We analyse the decrease in Finland's TFR in 2010-17, and assess its consequences for cohort fertility using complementary approaches. Decomposition of this fertility decline shows that first births and women aged <30 are making the largest contributions. However, women aged 30-39 are also, for the first time in decades, experiencing a sustained fertility decline. Tempo adjustments to the TFR suggest that quantum change is part of the decline. Several forecasting methods indicate that cohort fertility is likely to decline from the long-lasting level of 1.85-1.95 to 1.75 or lower among women born in the mid-1980s. Without an exceptionally strong recovery in fertility, Finnish cohort fertility is likely to decline to levels currently observed among countries with very low fertility.


The extreme C terminus of the Pseudomonas aeruginosa effector ExoY is crucial for binding to its eukaryotic activator, F-actin.
Bacterial nucleotidyl cyclase toxins are potent virulence factors that upon entry into eukaryotic cells are stimulated by endogenous cofactors to catalyze the production of large amounts of 3'5'-cyclic nucleoside monophosphates. The activity of the effector ExoY from Pseudomonas aeruginosa is stimulated by the filamentous form of actin (F-actin). Utilizing yeast phenotype analysis, site-directed mutagenesis, functional biochemical assays, and confocal microscopy, we demonstrate that the last nine amino acids of the C terminus of ExoY are crucial for the interaction with F-actin and, consequently, for ExoY's enzymatic activity in vitro and toxicity in a yeast model. We observed that isolated C-terminal sequences of P. aeruginosa ExoY that had been fused to a carrier protein bind to F-actin and that synthetic peptides corresponding to the extreme ExoY C terminus inhibit ExoY enzymatic activity in vitro and compete with the full-length enzyme for F-actin binding. Interestingly, we noted that various P. aeruginosa isolates of the PA14 family, including highly virulent strains, harbor ExoY variants with a mutation altering the C terminus of this effector. We found that these naturally occurring ExoY variants display drastically reduced enzymatic activity and toxicity. Our findings shed light on the molecular basis of the ExoY-F-actin interaction, revealing that the extreme C terminus of ExoY is critical for binding to F-actin in target cells and that some P. aeruginosa isolates carry C-terminally mutated, low-activity ExoY variants.


Effect of soymilk consumption on serum estrogen and androgen concentrations in Japanese men.
Soy consumption has been associated with a reduced risk of prostate cancer. The mechanism for this association may involve the effect of soy on the endocrine system. We conducted a randomized dietary intervention study to determine the effects of soy consumption on serum levels of steroid hormones in men. Thirty-five men were randomly assigned to either a soymilk-supplemented group or a control group. The men in the soy-supplemented group were asked to consume 400 ml of soymilk daily for 8 weeks. The men in the control group maintained their usual diet. Blood samples were obtained just before the initiation of the dietary period and thereafter every two weeks for 12 weeks. Changes in hormone concentrations were analyzed and compared between the two groups using the mixed linear regression model against weeks from the start of the dietary period. The mean (SD) soymilk intake estimated from dietary records during the dietary study period was 342.9 (SD, 74.2) ml in the soymilk-supplemented group. There was a significant difference between the two groups in terms of changes in serum estrone concentrations, which tended to decrease in the soy-supplemented group and increase in the control group over time. None of the other hormones measured (estradiol, total and free-testosterone, or sex hormone-binding globulin) showed any statistical difference between the two groups in terms of patterns of change. The results of the study indicate that soymilk consumption may modify circulating estrone concentrations in men.


Interfacial mixing in heteroepitaxial growth.
We investigate the growth of a film of some element B on a substrate made of another substance A in a model of molecular beam epitaxy. A vertical exchange mechanism (partial surfactant behavior) allows the A atoms to stay on the growing surface with a certain probability. Using kinetic Monte Carlo simulations as well as scaling arguments, the incorporation of the A 's into the growing B layer is investigated. Moreover, we develop a rate equation theory for this process. The concentration of A impurities decays in the B -film like (distance from the interface)(-1-beta), where beta approximately 0.5 for two-dimensional surfaces, approximately 0.8 in the one-dimensional case, and 1 in mean-field approximation. The power law is cut off exponentially at a characteristic thickness of the interdiffusion zone that depends on the rate of exchange of a B adatom with an A atom in the surface and on the diffusion length. Under certain conditions the interdiffusion zone is predicted to become narrower, if the growth temperature is increased.


The breeding biology of lemon sharks at a tropical nursery lagoon.
Surprisingly little is known about the reproductive behaviour and breeding biology of most shark species, especially in natural populations. Here, we characterize reproductive patterns and use of a natal nursery at Bimini, Bahamas by lemon sharks, Negaprion brevirostris. We systematically and exhaustively sampled young lemon sharks at Bimini annually from 1995 to 2000 and opportunistically sampled adults over the same period. Out of the 897 young sharks sampled, 119 could be assigned to five sampled mothers using microsatellite genotyping. Reproductive females showed strong philopatry to the nursery, returning to Bimini every two years to give birth. Each of these females may rely entirely on the Bimini nursery for recruitment. The protection of known nursery grounds should therefore figure prominently in conservation efforts for large coastal shark species. The reconstruction of paternal genotypes indicates that litters are sired by multiple males, and females mate with different males nearly every breeding cycle. The ubiquitous polyandry reported here raises the possibility that genetic incompatibility and post-copulatory paternity-biasing mechanisms may operate in viviparous sharks.


Integration of EGFR and LIN-12/Notch Signaling by LIN-1/Elk1, the Cdk8 Kinase Module, and SUR-2/Med23 in Vulval Precursor Cell Fate Patterning in Caenorhabditis elegans.
Six initially equivalent, multipotential Vulval Precursor Cells (VPCs) in Caenorhabditis elegans adopt distinct cell fates in a precise spatial pattern, with each fate associated with transcription of different target genes. The pattern is centered on a cell that adopts the "1°" fate through Epidermal Growth Factor Receptor (EGFR) activity, and produces a lateral signal composed of ligands that activate LIN-12/Notch in the two flanking VPCs to cause them to adopt "2°" fate. Here, we investigate orthologs of a transcription complex that acts in mammalian EGFR signaling-lin-1/Elk1, sur-2/Med23, and the Cdk8 Kinase module (CKM)-previously implicated in aspects of 1° fate in C. elegans and show they act in different combinations for different processes for 2° fate. When EGFR is inactive, the CKM, but not SUR-2, helps to set a threshold for LIN-12/Notch activity in all VPCs. When EGFR is active, all three factors act to resist LIN-12/Notch, as revealed by the reduced ability of ectopically-activated LIN-12/Notch to activate target gene reporters. We show that overcoming this resistance in the 1° VPC leads to repression of lateral signal gene reporters, suggesting that resistance to LIN-12/Notch helps ensure that P6.p becomes a robust source of the lateral signal. In addition, we show that sur-2/Med23 and lin-1/Elk1, and not the CKM, are required to promote endocytic downregulation of LIN-12-GFP in the 1° VPC. Finally, our analysis using cell fate reporters reveals that both EGFR and LIN-12/Notch signal transduction pathways are active in all VPCs in lin-1/Elk1 mutants, and that lin-1/Elk1 is important for integrating EGFR and lin-12/Notch signaling inputs in the VPCs so that the proper gene complement is transcribed.


Realignment of the ventricular septum using partial direct closure of the ventricular septal defect in Tetralogy of Fallot.
The aim is to describe our technique of partial direct closure of the ventricular septal defect (VSD) in Tetralogy of Fallot (TOF), and assess its influence on the realignment and remodeling of the left ventricular outflow tract. Between 2004 and 2010, 32 non-consecutive patients with TOF underwent a direct or partial direct closure of VSD. Median age and weight were 5.2 months and 6.7 kg, respectively. An approach through the right atrium was used in 30 patients and through the infundibulum in two patients. The conal septum was mobilized by transecting the hypertrophic trabeculae to facilitate the approximation of the VSD. The membranous part of the VSD was closed (in the later part of the series) with a small xenopericardial patch to avoid tension on the suture line traversing the area of risk to the bundle of His. Follow-up was complete, with a median duration of 46.9 (range 12-75.3) months. The VSD could be closed successfully in all patients. A residual VSD was partly responsible for one early postoperative re-operation. There were no early or late deaths. At follow-up, all patients were in sinus rhythm. Three patients showed a small residual VSD. Thirty patients had none, one showed trivial, and one had mild aortic regurgitation. The left ventricular outflow showed a good realignment of the ventricular septum in all the patients. Partial direct closure of the VSD corrects the primary defect in TOF, that is, the malalignment of the septum. It results in a straight, wide open left ventricular outflow tract and brings better support to the aortic root.


All-cause mortality benefit of coronary revascularization vs. medical therapy in patients without known coronary artery disease undergoing coronary computed tomographic angiography: results from CONFIRM (COronary CT Angiography EvaluatioN For Clinical Outcomes: An InteRnational Multicenter Registry).
To date, the therapeutic benefit of revascularization vs. medical therapy for stable individuals undergoing invasive coronary angiography (ICA) based upon coronary computed tomographic angiography (CCTA) findings has not been examined. We examined 15 223 patients without known coronary artery disease (CAD) undergoing CCTA from eight sites and six countries who were followed for median 2.1 years (interquartile range 1.4-3.3 years) for an endpoint of all-cause mortality. Obstructive CAD by CCTA was defined as a ≥50% luminal diameter stenosis in a major coronary artery. Patients were categorized as having high-risk CAD vs. non-high-risk CAD, with the former including patients with at least obstructive two-vessel CAD with proximal left anterior descending artery involvement, three-vessel CAD, and left main CAD. Death occurred in 185 (1.2%) patients. Patients were categorized into two treatment groups: revascularization (n = 1103; 2.2% mortality) and medical therapy (n = 14 120, 1.1% mortality). To account for non-randomized referral to revascularization, we created a propensity score developed by logistic regression to identify variables that influenced the decision to refer to revascularization. Within this model (C index 0.92, χ2 = 1248, P < 0.0001), obstructive CAD was the most influential factor for referral, followed by an interaction of obstructive CAD with pre-test likelihood of CAD (P = 0.0344). Within CCTA CAD groups, rates of revascularization increased from 3.8% for non-high-risk CAD to 51.2% high-risk CAD. In multivariable models, when compared with medical therapy, revascularization was associated with a survival advantage for patients with high-risk CAD [hazards ratio (HR) 0.38, 95% confidence interval 0.18-0.83], with no difference in survival for patients with non-high-risk CAD (HR 3.24, 95% CI 0.76-13.89) (P-value for interaction = 0.03). In an intermediate-term follow-up, coronary revascularization is associated with a survival benefit in patients with high-risk CAD by CCTA, with no apparent benefit of revascularization in patients with lesser forms of CAD.


Long-term consumption of Moringa oleifera-supplemented diet enhanced neurocognition, suppressed oxidative stress, acetylcholinesterase activity and neuronal degeneration in rat's hippocampus.
This study investigates protection against oxidative stress and memory enhancing potential of long-term consumption of Moringa oleifera leaves. Male Wistar rat were fed with mixture of M. oleifera-supplemented diets (MOSD) partitioned in 1, 5, 10, and 20% continuously for 12 weeks. Object recognition test (ORT) and Morris water maze (MWM) was used for assessing neurocognition. Changes in body weight, Lipid peroxidation (MDA), Glutathione (GSH), Catalase (CAT) and Acetylcholinesterase (AChE) activity was assayed in the brain tissue. Histomorphometric of the hippocampus was also examined. The diets progressively increase the body weigh after the 12 weeks, improved spatial (MWM) and non-spatial (ORT) memory performance, protect against oxidative stress, inhibit AChE activity and suppresses neuronal degeneration in the hippocampus when stained with Cresyl violent stain. Conclusively, long-term consumption of MOSD shows strong protection against oxidative stress and hippocampal degeneration and improves neurocognition with dose dependent effect.


Direct electrospinning of poly(vinyl butyral) onto human dermal fibroblasts using a portable device.
To demonstrate that uniform poly(vinyl butyral) (PVB) fibres can be safely electrospun onto a monolayer of human dermal fibroblasts using a portable device. PVB in solvent mixtures containing various amounts of ethanol and water was electrospun. Six percent (weight-to-volume ratio) PVB in a 9:1 ethanol:water ratio was the solution with the highest content in water that could be electrospun into consistent fibres with an average diameter of 0.9 μm (± 0.1 μm). Four and five percent PVB solutions created beaded fibres. A 8:2 ethanol:water solution lead to microbead formation while a 7:3 ethanol:water mix failed to fully dissolve. The selected solution was successfully electrospun onto a monolayer of human dermal fibroblasts and the process had no significant effect (p < 0.05) on cell viability compared to the control without fibres. PVB-ethanol-water solutions could be electrospun without damaging the exposed cell layer. However, further work is required to demonstrate the long-term effect of PVB as a wound healing material.


How did New Zealand's regional District Health Board groupings work to improve service integration and health outcomes: a realist evaluation.
In Aotearoa New Zealand (NZ), integration across the healthcare continuum has been a key approach to strengthening the health system and improving health outcomes. A key example has been four regional District Health Board (DHB) groupings, which, from 2011 to 2022, required the country's 20 DHBs to work together regionally. This research explores how this initiative functioned, examining how, for whom and in what circumstances regional DHB groupings worked to deliver improvements in system integration and health outcomes and equity. We used a realist-informed evaluation study design. We used documentary analysis to develop programme logic models to describe the context, structure, capabilities, implementation activities and impact of each of the four regional groupings and then conducted interviews with stakeholders. We developed a generalised context-mechanisms-outcomes model, identifying key commonalities explaining how regional work 'worked' across NZ while noting important regional differences. NZ's four regional DHB groupings. Forty-nine stakeholders from across the four regional groupings. These included regional DHB governance groups and coordinating regional agencies, DHB senior leadership, Māori and Pasifika leadership and lead clinicians for regional work streams. Regional DHB working was layered on top of an already complex DHB environment. Organisational heterogeneity and tensions between local and regional priorities were key contextual factors. In response, regional DHB groupings leveraged a combination of 'hard' policy and planning processes, as well as 'soft', relationship-based mechanisms, aiming to improve system integration, population health outcomes and health equity. The complexity of DHB regional working meant that success hinged on building relationships, leadership and trust, alongside robust planning and process mechanisms. As NZ reorients its health system towards a more centralised model underpinned by collaborations between local providers, our findings point to a need to align policy expectations and foster environments that support connection and collegiality across the health system.


[Principles of cognitive remediation in schizophrenia].
Cognitive remediation is an innovative psychosocial therapy which can provide a substantial benefit, especially for schizophrenic patients. As its name implies, the aim of cognitive remediation is to restore cognitive functions. Most cognitive domains (attention, memory and executive functions) are impaired in schizophrenia. Remediation therapy must be administered by an expert, and is based on cognitive training on the one hand, and on learning of cognitive strategies on the other hand. With these techniques the patient is better able to solve complex cognitive problems and to apply these new skills to everyday situations. Several techniques are available in France, using either computer-based or paper/pencil approaches. The programs are administered over several months, with one or more sessions per week. Cognitive remediation itself provides only a modest cognitive benefit, which must be enhanced by the adjunction of other therapies such as behavioral therapy, learning of social skills, or a vocational program during the first months of employment.


An oncogenic epidermal growth factor receptor signals via a p21-activated kinase-caldesmon-myosin phosphotyrosine complex.
Many ligand-independent receptor tyrosine kinases are tumorigenic. The biochemical signals that mediate ligand-independent transformation of cells by these transmembrane receptors are poorly defined. In this report, we demonstrate that a constitutively activated mutant epidermal growth factor receptor (v-ErbB) induces the formation of a transformation-specific signaling module that complexes with myosin II. The components of this signaling complex include the signal adapter proteins Shc, Grb2, and Nck, and tyrosine-phosphorylated forms of p21-activated kinase (Pak), caldesmon, and myosin light chain kinase. Transformation-specific, tyrosine phosphorylation of Pak enhances the catalytic activity of this serine/threonine kinase. Furthermore, the tyrosine phosphorylation of Pak is Rho-, but not Ras-, Rac-, or Cdc42-dependent. These results demonstrate that a ligand-independent epidermal growth factor receptor mutant can transduce oncogenic signals that are distinct from ligand-dependent, mitogenic signals. In addition, these data provide evidence for the coupling of oncogenic receptor tyrosine kinases with the actomyosin molecular motor. This myosin-associated signaling module may mediate some of the biochemical changes of myosin found in v-ErbB- transformed fibroblasts, thereby contributing to the regulation of the mechanical forces governing cellular adhesion, cytoskeletal tension, and, hence, anchorage-independent cell growth.


Transfer and Redistribution of Salmonella Typhimurium LT2 and Escherichia coli O157:H7 during Pilot-Scale Processing of Baby Spinach, Cilantro, and Romaine Lettuce.
Several outbreaks of foodborne illness traced to leafy greens and culinary herbs have been hypothesized to involve cross-contamination during washing and processing. This study aimed to assess the redistribution of Salmonella Typhimurium LT2 during pilot-scale production of baby spinach and cilantro and redistribution of Escherichia coli O157:H7 during pilot-scale production of romaine lettuce. Four inoculated surrogate:uninoculated product weight ratios (10:100, 5:100, 1:100, and 0.5:100) and three inoculation levels (103, 101, and 10-1 CFU/g) were used for the three commodities. For each of three trials per condition, 5-kg batches containing uninoculated product and spot-inoculated surrogate products at each ratio and inoculation level were washed for 90 s in a 3.6-m-long flume tank through which 890 L of sanitizer-free, filtered tap water was circulated. After washing and removing the inoculated surrogate products, washed product (∼23, 225-g samples per trial) was analyzed for presence or absence of Salmonella Typhimurium or E. coli O157:H7 by using the GeneQuence Assay. For baby spinach, cilantro, and romaine lettuce, no significant differences ( P > 0.05) in the percentage of positive samples were observed at the same inoculation level and inoculated:uninoculated weight ratio. For each pathogen product evaluated (triplicate trials), inoculation level had a significant impact on the percentage of positive samples after processing, with the percentage of positive samples decreasing, as the initial surrogate inoculation level decreased. The weight ratio of contaminated:noncontaminated product plays an important role: positive samples ranged from 0% to 11.6% ± 2.05% and from 68.1% ± 33.6% to 100% among the four ratios at inoculation of 10-1 and 101 CFU/g, respectively. To our knowledge, this study is the first to assess the redistribution of low levels of pathogens from incoming product to leafy greens during processing and should provide important data for microbial risk assessments and other types of food safety analyses related to fresh-cut leafy greens.


Fixed subaortic stenosis: anatomical spectrum and nature of progression.
Retrospective echocardiographic review identified 58 consecutive infants and children with fixed subaortic stenosis. Mean (SD) age at diagnosis was 4.8 (3.6) years (range two days to 14.7 years), and diagnosis occurred in infancy in eight. Associated cardiac abnormalities were present in 41 (71%) whereas fixed subaortic stenosis was an isolated lesion in 17 (29%). Four types of fixed subaortic stenosis were identified: short segment (47 (81%)), long segment (7 (12%)), posterior displacement of the infundibular septum with additional discrete narrowing of the left ventricular outflow tract (3 (5%)), and redundant tissue arising from the membranous septum (1 (2%)). Echocardiographic studies had been performed before the diagnosis of fixed subaortic stenosis in nine patients, all with associated abnormalities. These were performed in infancy in each and showed a "normal" left ventricular outflow tract in six and posterior deviation of the infundibular septum in three. In 16 patients serial echocardiographic studies had been performed after the diagnosis of fixed subaortic stenosis but before surgery of the left ventricular outflow tract. Rapid evolution of short segment to long segment narrowing was seen in one patient, and tethering of the aortic valve or mitral valve developed in a further four patients. Aortic valve or mitral valve involvement was not seen before the age of three years but was common thereafter (10/40 patients, 25%). Fixed subaortic stenosis may be an "acquired" lesion with the potential for changes in form as well as progression in severity of left ventricular outflow tract obstruction.


Bioactive cytokinins are selectively secreted by Sinorhizobium meliloti nodulating and nonnodulating strains.
Bacteria present in the rhizosphere of plants often synthesize phytohormones, and these signals can consequently affect root system development. In legumes, plants adapt to nitrogen starvation by forming lateral roots as well as a new organ, the root nodule, following a symbiotic interaction with bacteria collectively referred to as rhizobia. As cytokinin (CK) phytohormones were shown to be necessary and sufficient to induce root nodule organogenesis, the relevance of CK production by symbiotic rhizobia was questioned. In this study, we analyzed quantitatively, by liquid chromatography-tandem mass spectrometry, the production of 25 forms of CK in nine rhizobia strains belonging to four different species. All bacterial strains were able to synthesize a mix of CK, and bioactive forms of CK, such as iP, were notably found to be secreted in bacterial culture supernatants. Use of a mutant affected in extracellular polysaccharide (EPS) production revealed a negative correlation of EPS production with the ability to secrete CK. In addition, analysis of a nonnodulating Sinorhizobium meliloti strain revealed a similar pattern of CK production and secretion when compared with a related nodulating strain. This indicates that bacterially produced CK are not sufficient to induce symbiotic nodulation.


Effect of specific exercise-based football injury prevention programmes on the overall injury rate in football: a systematic review and meta-analysis of the FIFA 11 and 11+ programmes.
To investigate the effect of FIFA injury prevention programmes in football (FIFA 11 and FIFA 11+). Systematic review and meta-analysis. Randomised controlled trials comparing the FIFA injury prevention programmes with a control (no or sham intervention) among football players. MEDLINE via PubMed, EMBASE via OVID, CINAHL via Ebsco, Web of Science, SportDiscus and Cochrane Central Register of Controlled Trials, from 2004 to 14 March 2016. 6 cluster-randomised controlled trials had assessed the effect of FIFA injury prevention programmes compared with controls on the overall football injury incidence in recreational/subelite football. These studies included 2 specific exercise-based injury prevention programmes: FIFA 11 (2 studies) and FIFA 11+ (4 studies). The primary analysis showed a reduction in the overall injury risk ratio of 0.75 (95% CI 0.57 to 0.98), p=0.04, in favour of the FIFA injury prevention programmes. Secondary analyses revealed that when pooling the 4 studies applying the FIFA 11+ prevention programme, a reduction in the overall injury risk ratio (incidence rate ratio (IRR) 0.61; 95% CI 0.48 to 0.77, p<0.001) was present in favour of the FIFA 11+ prevention programme. No reduction was present when pooling the 2 studies including the FIFA 11 prevention programme (IRR 0.99; 95% CI 0.80 to 1.23, p=0.940). An injury-preventing effect of the FIFA injury prevention programmes compared with controls was shown in football. This effect was induced by the FIFA 11+ prevention programme which has a substantial injury-preventing effect by reducing football injuries by 39%, whereas a preventive effect of the FIFA 11 prevention programme could not be documented. PROSPERO CRD42015024120.


A nonrandom chromosomal abnormality, del 3p(14-23), in human small cell lung cancer (SCLC).
In order to determine whether or not there are specific chromosomal changes in small cell lung cancer (SCLC), karyotypic analyses of 16 continuous SCLC tissue culture lines, three fresh tumor specimens (bone marrow), one direct preparation of bone marrow involved with SCLC, and two lymphoblastoid lines derived from SCLC patients were studied. Cell lines were derived from primary tumor, or metastases to bone marrow, subcutaneous nodules, or pleural fluid; all 16 lines had biochemical and histologic properties characteristic of SCLC. Of the 15 males and 3 females, 6 patients had no prior treatment. All of the 16 cell lines, the 3 fresh specimens, and the direct bone marrow preparation had a common deletion of the short arm of chromosome #3. Use of the shortest region of overlap analysis showed the common deletion was of the short arm in the regions p(14-23). This specific chromosomal abnormality, del 3p, was not found in five non-SCLC cell lines studied and is of major potential biological and diagnostic importance.


The fatty acid synthase inhibitor orlistat reduces the growth and metastasis of orthotopic tongue oral squamous cell carcinomas.
Fatty acid synthase (FASN) is the biosynthetic enzyme responsible for the endogenous synthesis of fatty acids. It is downregulated in most normal cells, except in lipogenic tissues such as liver, lactating breast, fetal lung, and adipose tissue. Conversely, several human cancers, including head and neck squamous cell carcinomas (HNSCC), overexpress FASN, which has been associated with poor prognosis and recently suggested as a metabolic oncoprotein. Orlistat is an irreversible inhibitor of FASN activity with cytotoxic properties on several cancer cell lines that inhibits tumor progression and metastasis in prostate cancer xenografts and experimental melanomas, respectively. To explore whether the inhibition of FASN could impact oral tongue squamous cell carcinoma (OTSCC) metastatic spread, an orthotopic model was developed by the implantation of SCC-9 ZsGreen LN-1 cells into the tongue of BALB/c nude mice. These cells were isolated through in vivo selection, show a more invasive behavior in vitro than the parental cells, and generate orthotopic tumors that spontaneously metastasize to cervical lymph nodes in 10 to 15 days only. SCC-9 ZsGreen LN-1 cells also exhibit enhanced production of MMP-2, ERBB2, and CDH2. The treatment with orlistat reduced proliferation and migration, promoted apoptosis, and stimulated the secretion of VEGFA165b by SCC-9 ZsGreen LN-1 cells. In vivo, the drug was able to decrease both the volume and proliferation indexes of the tongue orthotopic tumors and, importantly, reduced the number of metastatic cervical lymph nodes by 43%. These results suggest that FASN is a potential molecular target for the chemotherapy of patients with OTSCC.


Association of Active and Sedentary Behaviors with Postmenopausal Estrogen Metabolism.
Physical activity may reduce endogenous estrogens, but few studies have assessed effects on estrogen metabolism and none have evaluated sedentary behavior in relation to estrogen metabolism. We assessed relationships between accelerometer-measured physical activity and sedentary behavior and 15 urinary estrogens and estrogen metabolites (EM) among postmenopausal controls from a population-based breast cancer case-control study conducted in Poland (2000-2003). Postmenopausal women (N = 542) were ages 40 to 72 yr and not currently using hormone therapy. Accelerometers, worn for 7 d, were used to derive measures of average activity (counts per day) and sedentary behavior (<100 counts per minute per day). Estrogen metabolites were measured in 12-h urine samples using liquid chromatography-tandem mass spectrometry. Estrogen metabolites were analyzed individually, in metabolic pathways (C-2, -4, or -16), and as ratios relative to parent estrogens. Geometric means of estrogen metabolites by tertiles of accelerometer-measures, adjusted for age and body mass, were computed using linear models. High activity was associated with lower levels of estrone and estradiol (P trend = 0.01), whereas increased sedentary time was positively associated with these parent estrogens (P trend = 0.04). Inverse associations were observed between high activity and 2-methoxyestradiol, 4-methoxyestradiol, 17-epiestriol, and 16-epiestriol (P trend = 0.03). Sedentary time was positively associated with methylated catechols in the 2- and 4-hydroxylation pathways (P trend ≤ 0.04). Women in the highest tertile of activity had increased hydroxylation at the C-2, -4, and -16 sites relative to parent estrogens (P trend ≤ 0.02), whereas increased sedentary time was associated with a lower 16-pathway/parent estrogen ratio (P trend = 0.01). Higher activity was associated with lower urinary estrogens, possibly through increased estrogen hydroxylation and subsequent metabolism, whereas sedentary behavior may reduce metabolism.


When Good Bugs Go Bad: Epidemiology and Antimicrobial Resistance Profiles of Corynebacterium striatum, an Emerging Multidrug-Resistant, Opportunistic Pathogen.
Infections with Corynebacterium striatum have been described in the literature over the last 2 decades, with the majority being bacteremia, central line infections, and occasionally, endocarditis. In recent years, the frequency of C. striatum infections appears to be increasing; a factor likely contributing to this is the increased ease and accuracy of the identification of Corynebacterium spp., including C. striatum, from clinical cultures. The objective of this study was to retrospectively characterize C. striatum isolates recovered from specimens submitted as part of routine patient care at a 1,250-bed, tertiary-care academic medical center. Multiple strain types were recovered, as demonstrated by repetitive-sequence-based PCR. Most of the strains of C. striatum characterized were resistant to antimicrobials commonly used to treat Gram-positive organisms, such as penicillin, ceftriaxone, meropenem, clindamycin, and tetracycline. The MIC50 for ceftaroline was >32 μg/ml. Although there are no interpretive criteria for susceptibility with telavancin, it appeared to have potent in vitro efficacy against this species, with MIC50 and MIC90 values of 0.064 and 0.125 μg/ml, respectively. Finally, as previously reported in case studies, we demonstrated rapid in vitro development of daptomycin resistance in 100% of the isolates tested (n = 50), indicating that caution should be exhibited when using daptomycin for the treatment of C. striatum infections. C. striatum is an emerging, multidrug-resistant pathogen that can be associated with a variety of infection types.


Variants in the 3'-untranslated region of CUL3 is associated with risk of esophageal squamous cell carcinoma.
Background: Esophageal squamous cell carcinoma (ESCC) is one of the most lethal cancers in China. Recently, a study identified that cullin 3 (CUL3) was significantly mutated and deleted in ESCC. We then hypothesis that germline variants in CUL3 may also associated with the susceptibility of ESCC. Variants in the gene 3'-untranslated region (3'-UTR) may associate with gene expression by altering miRNAs binding. Material and Methods: We systematically searched for variants in the 3'-UTR of CUL3 using the Ensembl database. Taqman SNP Genotyping Assay was performed in 638 ESCC cases and 546 controls to examine the association between the rs2396092 and the risk of ESCC. The eQTL analysis for CUL3 were conducted by using the GTEx database. Results: We identified that the rs2396092 was significantly associated with the susceptibility of ESCC. Compared with the TT genotype carriers, the CT genotype and CC genotype carriers were correlated with risk of ESCC with odds ratio being 1.33 (95% CI: 1.04-1.70, P=0.0222) and 1.63 (95% CI: 1.07-2.50, P=0.0241), respectively. Different genotypes of rs2396092 was also shown to be correlated with altered CUL3 expression. Conclusion: The results emphasize the importance of CUL3 in the development of ESCC and may contribute to the personalized prevention of this cancer in the future.


Rapid response of a marine mammal species to holocene climate and habitat change.
Environmental change drives demographic and evolutionary processes that determine diversity within and among species. Tracking these processes during periods of change reveals mechanisms for the establishment of populations and provides predictive data on response to potential future impacts, including those caused by anthropogenic climate change. Here we show how a highly mobile marine species responded to the gain and loss of new breeding habitat. Southern elephant seal, Mirounga leonina, remains were found along the Victoria Land Coast (VLC) in the Ross Sea, Antarctica, 2,500 km from the nearest extant breeding site on Macquarie Island (MQ). This habitat was released after retreat of the grounded ice sheet in the Ross Sea Embayment 7,500-8,000 cal YBP, and is within the range of modern foraging excursions from the MQ colony. Using ancient mtDNA and coalescent models, we tracked the population dynamics of the now extinct VLC colony and the connectivity between this and extant breeding sites. We found a clear expansion signal in the VLC population approximately 8,000 YBP, followed by directional migration away from VLC and the loss of diversity at approximately 1,000 YBP, when sea ice is thought to have expanded. Our data suggest that VLC seals came initially from MQ and that some returned there once the VLC habitat was lost, approximately 7,000 years later. We track the founder-extinction dynamics of a population from inception to extinction in the context of Holocene climate change and present evidence that an unexpectedly diverse, differentiated breeding population was founded from a distant source population soon after habitat became available.


Experiences in end-of-life care in the Intensive Care Unit: A survey of resident physicians.
The practice of intensive care includes withholding and withdrawal of care, when appropriate, and the goals of care change around this time to comfort and palliation. We decided to survey the attitudes, training, and skills of intensive care residents in relation to end-of-life (EoL) care. All residents at our institute who has worked for at least a month in an adult Intensive Care Unit were invited to participate. After Institutional Ethics Committee approval, a Likert-scale questionnaire, divided into five composite measures of EoL skills including training and attitude, was handed over to individual residents and completed data were anonymized. Frequency and descriptive analysis was performed for the demographic variables. Central tendency, variability, and reliability were examined for the five composite measures. Scale internal consistency was checked by Cronbach's coefficient alpha. Multivariate forward conditional regression analysis was conducted to examine the association of demographic data or EoL experience to composite measures. Of the 170 eligible residents, we received 120 (70.5%) responses. Internal medicine residents have more experience in caring for dying patients and conducting EoL discussions. Even though majority of participants reported that they are comfortable with the concept of EoL care, this does not always reflect the actual practice in the hospital. There is a need for further training in skills around EoL care. As this is a self-assessment survey, the specific measures of attitudes and skills in EoL are poorly reflected, indicating a need for further research.


Characterization of Posa and Posa-like virus genomes in fecal samples from humans, pigs, rats, and bats collected from a single location in Vietnam.
Porcine stool-associated RNA virus (posavirus), and Human stool-associated RNA virus (husavirus) are viruses in the order Picornavirales recently described in porcine and human fecal samples. The tentative group (Posa and Posa-like viruses: PPLVs) also includes fish stool-associated RNA virus (fisavirus) as well as members detected in insects (Drosophila subobscura and Anopheles sinensis) and parasites (Ascaris suum). As part of an agnostic deep sequencing survey of animal and human viruses in Vietnam, we detected three husaviruses in human fecal samples, two of which share 97-98% amino acid identity to Dutch husavirus strains and one highly divergent husavirus with only 25% amino acid identity to known husaviruses. In addition, the current study found forty-seven complete posavirus genomes from pigs, ten novel rat stool-associated RNA virus genomes (tentatively named rasavirus), and sixteen novel bat stool-associated RNA virus genomes (tentatively named basavirus). The five expected Picornavirales protein domains (helicase, 3C-protease, RNA-dependent RNA polymerase, and two Picornavirus capsid domain) were found to be encoded by all PPLV genomes. In addition, a nucleotide composition analysis revealed that the PPLVs shared compositional properties with arthropod viruses and predicted non-mammalian hosts for all PPLV lineages. The study adds seventy-six genomes to the twenty-nine PPLV genomes currently available and greatly extends our sequence knowledge of this group of viruses within the Picornavirales order.


Preventive home visits and health--experiences among very old people.
As more people reach older age, there is a growing interest in improving old person's health, activity, independence and social participation, thereby adding quality to the extended years. Preventive home visits (PHV) programs for old people have received much attention in recent decades. A large body of research shows mixed effects, and argues that a home visit is a complex social process influenced by numerous factors. To evaluate the impact of PHV, as well as making decisions on whether, how, and to whom the service should be provided, requires a deeper understanding of PHV than we have now. Consequently, the aim of the study was to describe the variations in older people's (80+) experiences of a single preventive home visit and its consequences for health. Seventeen participants between 80 and 92 years of age who had all received a structured PHV were interviewed in their own homes. The interviews were analyzed using the phenomenographic method, looking at the variations in the participants' experiences. The interviews revealed four categories: "The PHV made me visible and proved my human value"; "The PHV brought a feeling of security"; "The PHV gave an incentive to action"; and "The PHV was not for me". The experiences of a PHV were twofold. On one hand, the positive experiences indicate that one structured PHV was able to empower the participants and strengthen their self-esteem, making them feel in control over their situation and more aware of the importance of keeping several steps ahead. Together this could motivate them to take measures and engage in health-promoting activities. On the other hand, the PHV was experienced as being of no value by a few. These findings may partly explain the positive results from PHV interventions and emphasize that one challenge for health care professionals is to motivate older people who are healthy and independent to engage in health-promoting and disease-preventive activities.


Low socioeconomic status associates with higher serum phosphate irrespective of race.
Hyperphosphatemia, which associates with adverse outcomes in CKD, is more common among blacks than whites for unclear reasons. Low socioeconomic status may explain this association because poverty both disproportionately affects racial and ethnic minorities and promotes excess intake of relatively inexpensive processed and fast foods enriched with highly absorbable phosphorus additives. We performed a cross-sectional analysis of race, socioeconomic status, and serum phosphate among 2879 participants in the Chronic Renal Insufficiency Cohort Study. Participants with the lowest incomes or who were unemployed had higher serum phosphate concentrations than participants with the highest incomes or who were employed (P < 0.001). Although we also observed differences in serum phosphate levels by race, income modified this relationship: Blacks had 0.11 to 0.13 mg/dl higher serum phosphate than whites in the highest income groups but there was no difference by race in the lowest income group. In addition, compared with whites with the highest income, both blacks and whites with the lowest incomes had more than twice the likelihood of hyperphosphatemia in multivariable-adjusted analysis. In conclusion, low socioeconomic status associates with higher serum phosphate concentrations irrespective of race. Given the association between higher levels of serum phosphate and cardiovascular disease, further studies will need to determine whether excess serum phosphate may explain disparities in kidney disease outcomes among minority populations and the poor.


Active-site-directed inactivation of wheat-germ aspartate transcarbamoylase by pyridoxal 5'-phosphate.
Treatment of 1 microM wheat-germ aspartate transcarbamoylase with 1 mM-pyridoxal 5'-phosphate caused a rapid loss of activity, concomitant with the formation of a Schiff base. Complete loss of activity occurred within 10 min when the Schiff base was reduced with a 100-fold excess of NaBH4. Concomitantly, one amino group per chain was modified. No further residues were modified in the ensuing 30 min. The kinetics of inactivation were examined under conditions where the Schiff base was reduced before assay. Inactivation was apparently first-order. The pseudo-first-order rate constant, kapp., showed a hyperbolic dependence upon the concentration of pyridoxal 5'-phosphate, suggesting that the enzyme first formed a non-covalent complex with the reagent, modification of a lysine then proceeding within this complex. Inactivation of the enzyme by pyridoxal was 20 times slower than that by pyridoxal 5'-phosphate, indicating that the phosphate group was important in forming the initial complex. Partial protection against pyridoxal phosphate was provided by the leading substrate, carbamoyl phosphate, and nearly complete protection was provided by the bisubstrate analogue, N-phosphonoacetyl-L-aspartate, and the ligand-pair carbamoyl phosphate plus succinate. Steady-state kinetic studies, under conditions that minimized inactivation, showed that pyridoxal 5'-phosphate was also a competitive inhibitor with respect to the leading substrate, carbamoyl phosphate. Pyridoxal 5'-phosphate therefore appears to be an active-site-directed reagent. A sample of the enzyme containing one reduced pyridoxyl group per chain was digested with trypsin, and the labelled peptide was isolated and shown to contain a single pyridoxyl-lysine residue. Partial sequencing around the labelled lysine showed little homology with the sequence surrounding lysine-84, an active-centre residue of the catalytic subunit of aspartate transcarbamoylase from Escherichia coli, whose reaction with pyridoxal 5'-phosphate shows many similarities to the results described in the present paper. Arguably the reactive lysine is conserved between the two enzymes whereas the residues immediately surrounding the lysine are not. The same conclusion has been drawn in a comparison of reactive histidine residues in the two enzymes [Cole & Yon (1986) Biochemistry 25, 7168-7174].


Protein kinase C isoform delta is involved in the stimulation of the Na(+)-H+ exchanger in C6 glioma cells.
Protein kinase C (PKC), the major receptor for tumor-promoting phorbol esters, is a family consisting of at least 12 distinct isoforms. In C6 glioma cells, short term (10 min) treatment with 12-O-tetradecanoyl phorbol-13-acetate (TPA) results in a dose-dependent translocation of PKC alpha, PKC delta and PKC theta. Long term (24 hr) treatment with appropriate doses of TPA results in the complete down-regulation of PKC delta but not of PKC alpha PKC theta. This property was used to determine which isoform might be involved in the activation of the glial cell Na(+)-H+ exchanger. It was found that (a) TPA has dose-dependent effects on PKC alpha, PKC delta, and PKC theta translocation and on Na(+)-dependent, EIPA-sensitive Na(+)-H+ exchanger stimulation; (b) the antiporter is blocked both by staurosporine and in cells in which PKC alpha, PKC delta, and PKC theta have been completely down-regulated; (c) the inactive form (alpha-TPA) of TPA, which does not include translocation of the three isoforms (alpha, delta, and theta) has no effect on the antiporter; (d) Western blot analysis demonstrated dose-dependent TPA (10, 30, 100, and 1000 nM)-induced translocation of PKC alpha, PKC delta, and PKC theta. TPA (10, 30, 100, and 1000 nM)-induced Na(+)-H+ exchanger activation was also found to be dose dependent. There was no difference in TPA-induced Na(+)-H+ exchanger activation between 30 and 100 nM; it correlated with the extent of TPA-induced PKC delta translocation over the same concentration range, suggesting that isoform responsible for the exchanger activation is PKC delta; (e) When 1 microM TPA is added after prior treatment of the cells with 10 nM TPA, an additive effect on Na(+)-H+ exchanger is seen that is not observed when the initial stimulus in 30 nM TPA, as would be expected if the PKC delta isoform were responsible for exchanger activation; (f) Finally, when PKC delta, but not PKC alpha and PKC theta, is completely down-regulated by 24-hr pretreatment with 10, 30, or 100 nM TPA, the Na(+)-H+ exchanger can no longer be stimulated by 1 microM TPA; when 1 nM TPA pretreatment is used, no down-regulation occurs and the exchanger still responds to 1 microM TPA. We have shown that the Na(+)-H+ exchanger in C6 glioma cells can be stimulated by TPA-induced PKC activation and, for the first time, that PKC delta is involved in the activation of this antiporter. Our results also suggest that different members of the PKC family within a single cell elicit specific physiological responses.


A population-based study showing better renal prognosis for proteinase 3 antineutrophil cytoplasmic antibody (ANCA)-associated nephritis versus myeloperoxidase ANCA-associated nephritis.
Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is usually differentiated based on clinical phenotypes, but recent data indicate that myeloperoxidase (MPO)-AAV is genetically distinct from proteinase 3 (PR3)-AAV. We reviewed a population-based cohort of AAV, focusing on differences in clinical and laboratory characteristics and to compare renal outcome between MPO-ANCA and PR3-ANCA nephritis. All new cases of AAV diagnosed between 1997 and 2009 in a geographically defined area in southern Sweden were retrieved using a validated search algorithm. Data were collected from time of diagnosis and end of followup. Renal and patient survival were analyzed according to ANCA serotype. During the study period, 201 patients were diagnosed with AAV, 98 tested positive for PR3-ANCA, and 85 for MPO-ANCA. Patients with PR3-ANCA were younger, had significantly higher inflammatory activity, and had a larger number of organs involved at diagnosis, but nephritis was more prevalent among patients with MPO-associated (72/85; 85%) versus PR3-associated disease (67/98, 68%). When comparing only patients with ANCA-associated nephritis, those with MPO-ANCA were more likely to develop endstage renal disease (n = 27, 38%) than those with PR3-ANCA (n = 10, 15%), p = 0.003. The risk remained significantly elevated after adjusting for sex, age, and s-creatinine level at diagnosis (HR 2.64; 95% CI 1.25-5.58; p = 0.003). There were no significant differences in mortality rates between the 2 groups. The outcome in this population-based cohort indicates that among AAV patients with nephritis, renal prognosis is better in the PR3-ANCA group, even after adjustment for sex, age, and renal function at diagnosis.