Enhanced Homing Technique of Mesenchymal Stem Cells Using Iron Oxide Nanoparticles by Magnetic Attraction in Olfactory-Injured Mouse Models.
Intranasal delivery of mesenchymal stem cells (MSCs) to the olfactory bulb is a promising approach for treating olfactory injury. Additionally, using the homing phenomenon of MSCs may be clinically applicable for developing therapeutic cell carriers. Herein, using superparamagnetic iron oxide nanoparticles (SPIONs) and a permanent magnet, we demonstrated an enhanced homing effect in an olfactory model. Superparamagnetic iron oxide nanoparticles with rhodamine B (IRBs) had a diameter of 5.22 ± 0.9 nm and ζ-potential of +15.2 ± 0.3 mV. IRB concentration of 15 µg/mL was injected with SPIONs into MSCs, as cell viability significantly decreased when 20 μg/mL was used (p ≤ 0.005) compared to in controls. The cells exhibited magnetic attraction in vitro. SPIONs also stimulated CXCR4 (C-X-C chemokine receptor type 4) expression and CXCR4-SDF-1 (Stromal cell-derived factor 1) signaling in MSCs. After injecting magnetized MSCs, these cells were detected in the damaged olfactory bulb one week after injury on one side, and there was a significant increase compared to when non-magnetized MSCs were injected. Our results suggest that SPIONs-labeled MSCs migrated to injured olfactory tissue through guidance with a permanent magnet, resulting in better homing effects of MSCs in vivo, and that iron oxide nanoparticles can be used for internalization, various biological applications, and regenerative studies.


Telescopic horizon scanning.
The problem of "distortionless" viewing with terrestrial telescopic systems (mainly "binoculars") remains problematic. The so called "globe effect" is only partially counteracted in modern designs. Theories addressing the phenomenon have never reached definitive closure. In this paper, we show that exact distortionless viewing with terrestrial telescopic systems is not possible in general, but that it is in principle possible in-very frequent in battle field and marine applications-the case of horizon scanning. However, this involves cylindrical optical elements. For opto-electronic systems, a full solution is more readily feasible. The solution involves a novel interpretation of the relevant constraints and objectives. For final design decisions, it is not necessary to rely on a corpus of psychophysical (or ergonomic) data, although one has to decide whether the instrument is intended as an extension of the eye or as a "pictorial" device.


A risk-adjusted and anatomically stratified cohort comparison study of open surgery, endovascular techniques and medical management for juxtarenal aortic aneurysms-the UK COMPlex AneurySm Study (UK-COMPASS): a study protocol.
In one-third of all abdominal aortic aneurysms (AAAs), the aneurysm neck is short (juxtarenal) or shows other adverse anatomical features rendering operations more complex, hazardous and expensive. Surgical options include open surgical repair and endovascular aneurysm repair (EVAR) techniques including fenestrated EVAR, EVAR with adjuncts (chimneys/endoanchors) and off-label standard EVAR. The aim of the UK COMPlex AneurySm Study (UK-COMPASS) is to answer the research question identified by the National Institute for Health Research Health Technology Assessment (NIHR HTA) Programme: 'What is the clinical and cost-effectiveness of strategies for the management of juxtarenal AAA, including fenestrated endovascular repair?' UK-COMPASS is a cohort study comparing clinical and cost-effectiveness of different strategies used to manage complex AAAs with stratification of physiological fitness and anatomical complexity, with statistical correction for baseline risk and indication biases. There are two data streams. First, a stream of routinely collected data from Hospital Episode Statistics and National Vascular Registry (NVR). Preoperative CT scans of all patients who underwent elective AAA repair in England between 1 November 2017 and 31 October 2019 are subjected to Corelab analysis to accurately identify and include every complex aneurysm treated. Second, a site-reported data stream regarding quality of life and treatment costs from prospectively recruited patients across England. Site recruitment also includes patients with complex aneurysms larger than 55 mm diameter in whom an operation is deferred (medical management). The primary outcome measure is perioperative all-cause mortality. Follow-up will be to a median of 5 years. The study has received full regulatory approvals from a Research Ethics Committee, the Confidentiality Advisory Group and the Health Research Authority. Data sharing agreements are in place with National Health Service Digital and the NVR. Dissemination will be via NIHR HTA reporting, peer-reviewed journals and conferences. ISRCTN85731188.


Comparative Genomic Reveals Clonal Heterogeneity in Persistent Staphylococcus aureus Infection.
Persistent infections caused by Staphylococcus aureus remain a clinical challenge. Adaptational mechanisms of the pathogen influencing infection persistence, treatment success, and clinical outcome in these types of infections by S. aureus have not been fully elucidated so far. We applied a whole-genome sequencing approach on fifteen isolates retrieved from a persistent S. aureus infection to determine their genetic relatedness, virulome, and resistome. The analysis of the genomic data indicates that all isolates shared a common clonal origin but displayed a heterogenous composition of virulence factors and antimicrobial resistance. This heterogeneity was reflected by different mutations in the rpoB gene that were related to the phenotypic antimicrobial resistance towards rifampicin and different minimal inhibitory concentrations of oxacillin. In addition, one group of isolates had acquired the genes encoding for staphylokinase (sak) and staphylococcal complement inhibitor (scn), leading to the truncation of the hemolysin b (hlb) gene. These features are characteristic for temperate phages of S. aureus that carry genes of the immune evasion cluster and confer triple conversion by integration into the hlb gene. Modulation of immune evasion mechanisms was demonstrated by significant differences in biofilm formation capacity, while invasion and intracellular survival in neutrophils were not uniformly altered by the presence of the immune evasion cluster. Virulence factors carried by temperate phages of S. aureus may contribute to the course of infection at different stages and affect immune evasion and pathogen persistence. In conclusion, the application of comparative genomic demonstrated clonal heterogeneity in persistent S. aureus infection.


Modulation of serotonin-induced vasospasm by endothelium and monoamine oxidase.
Endothelial modulation of flow induced by intraluminal serotonin (5-HT) in isolated and perfused bovine coronary artery segments was studied. A constant-pressure continuous perfusion apparatus was utilized. Control coronary arteries were perfused with a fixed volume of serotonin-containing solution followed by a serotonin-free solution, and flow-rate changes during onset and relaxation of vasospasm were measured. Both monoamine oxidase inhibition by iproniazide and endothelium disruption by collagenase increased the rate of onset and magnitude of vasospasm. When the endothelium was intact the vasospasm continued to increase, reaching maximum well after the end of the serotonin perfusion, followed by slow relaxation toward baseline. This contrasted with de-endothelialized vessels in which the increase in contractile response terminated abruptly at the end of the serotonin perfusion and returned rapidly to baseline. Coronary arteries stimulated with prostaglandin F2 alpha responded similarly to de-endothelialized vessels stimulated by 5-HT, although further de-endothelialization of F2 alpha-stimulated vessels showed increased rates of onset and relaxation of vasospasm, suggesting a physical barrier role for the endothelium towards unmetabolized agents. These observations are consistent with the hypothesis that endothelial cells are capable of taking up, storing and subsequently releasing serotonin. The results suggest a protective role of the endothelium as a metabolic and physical barrier. This may represent an anatomical substrate favouring the development of localized vasospasm at sites where the endothelium is injured.


Exercise as a therapeutic intervention in chronic kidney disease: are we nearly there yet?
The opportunity to review the more recent evidence for prescribing exercise-based physical rehabilitation for people living with chronic kidney disease (CKD) is timely. There has been a recent global focus evaluating how physical activity interventions might improve health-related quality of life and outcomes for people living with chronic health conditions in a post-COVID era. There is finally a long overdue commitment from the kidney research and clinical community to deliver pragmatic interventions to help people living with CKD to be able to live well with their condition. This article reviews recent research, and discusses the challenges and potential solutions, for providing exercise-based therapeutic options for people living with CKD; including predialysis self-management interventions, options for both prehabilitation and posttransplant rehabilitation, pragmatic considerations for delivery of exercise therapy for people receiving haemodialysis treatment and the role of virtual kidney-specific rehabilitation. Whilst there remains a need for further research in this area of patient care, there is now a body of evidence and kidney-specific guidelines that firmly support a rollout of pragmatic and scalable exercise-based interventions for people living with CKD. We are indeed nearly there now.


Regulation of hepatitis B virus mRNA expression in a hepatitis B surface antigen transgenic mouse model by IFN-gamma-secreting T cells after DNA-based immunization.
The immunotherapeutic effect of DNA-mediated immunization against chronic hepatitis B virus (HBV) infection has been evaluated in transgenic mice expressing the sequences that code for the envelope proteins of HBV in the liver. In this model of HBV chronic carriers, a single i.m. injection of plasmid DNA encoding HBV envelope proteins is sufficient to generate specific immune responses leading to the clearance of the transgene expression product and the control of HBV mRNA. The relative contributions of the T cell subpopulations induced by DNA immunization were examined using adoptive transfer experiments. It was shown that either CD8+ or CD4+ T lymphocytes from immunocompetent DNA-immunized animals were sufficient to control viral gene expression in the livers of the recipient transgenic mice. This effect was mediated by a cytokine-dependent mechanism common to both T cell subpopulations; this mechanism did not require cell lysis, but did involve the production of IFN-gamma by the activated T cells.


Participation of neural crest-derived cells in the genesis of the skull in birds.
The differentiation of cephalic neural crest cells into skeletal tissue in birds has been observed using the quail-chick nuclear marking system, which is based on specific differences in the distribution of the nuclear DNA. Chimaeras were formed by replacing a fragment of cephalic neural primordium of a 2- to 12-somite chicken embryo by the corresponding fragment isolated from an equivalent quail embryo. The participation of the graft-derived cells in the formation of the skull of these embryos was studied on histological sections after Feulgen and Rossenbeck staining. Cells from the prosencephalic neural crest migrate into the frontal nasal process and mix with the mesencephalic neural crest cells in the lateral nasal processes, around the optic cupule and beneath the diencephalon. In addition, the mesencephalic neural crest cells form the bulk of the mesenchyme of the maxillary processes and mandibular arch, whereas the rhombencephalic neural crest cells become located in the branchial arches. The origin of cartilages of the chondrocranium and bones of the neurocranium and viscerocranium has been shown in the chimaeric embryos: the basal plate cartilages, occipital bones, sphenoid bones and the cranial vault are mainly of mesodermal origin. However some parts have a dual origin: rhombo-mesencephalic neural crest cells are found in the otic capsule, and the frontal bone, the rostrum of parasphenoid and the orbital cartilages contain diverse amounts of prosencephalo-mesencephalic neural crest cells. The squamosals and the columella auris are formed from mesectodermic cells as are the nasal skeleton, the palatines and the maxillar bones. The mesectodermal origin of mandibular and hyoid bones and cartilages was already known. From these results it appears that the cephalic neural crest is particularly important in the formation of the facial part of the skull, while the vault and dorsal part are mesodermal and cartilages and bones found in the intermediary region are of mixed origin. The presence of mixed structures implies that the mesoderm and the mesectoderm are equally competent towards the specific inducers of these bones and cartilages. This correlates with the equivalence in differentiation capacities already shown for cephalic mesodermal and mesectodermal mesenchymes.


Translational relevance of SOS1 targeting for KRAS-mutant colorectal cancer.
It has been challenging to target mutant KRAS (mKRAS) in colorectal cancer (CRC) and other malignancies. Recent efforts have focused on developing inhibitors blocking molecules essential for KRAS activity. In this regard, SOS1 inhibition has arisen as an attractive approach for mKRAS CRC given its essential role as a guanine nucleotide exchange factor for this GTPase. Here, we demonstrated the translational value of SOS1 blockade in mKRAS CRC. We used CRC patient-derived organoids (PDOs) as preclinical models to evaluate their sensitivity to SOS1 inhibitor BI3406. A combination of in silico analyses and wet lab techniques was utilized to define potential predictive markers for SOS1 sensitivity and potential mechanisms of resistance in CRC. RNA-seq analysis of CRC PDOs revealed two groups of CRC PDOs with differential sensitivities to SOS1 inhibitor BI3406. The resistant group was enriched in gene sets involving cholesterol homeostasis, epithelial-mesenchymal transition, and TNF-α/NFκB signaling. Expression analysis identified a significant correlation between SOS1 and SOS2 mRNA levels (Spearman's ρ 0.56, p < 0.001). SOS1/2 protein expression was universally present with heterogeneous patterns in CRC cells but only minimal to none in surrounding nonmalignant cells. Only SOS1 protein expression was associated with worse survival in patients with RAS/RAF mutant CRC (p = 0.04). We also found that SOS1/SOS2 protein expression ratio >1 by immunohistochemistry (p = 0.03) instead of KRAS mutation (p = 1) was a better predictive marker to BI3406 sensitivity of CRC PDOs, concordant with the significant positive correlation between SOS1/SOS2 protein expression ratio and SOS1 dependency. Finally, we showed that GTP-bound RAS level underwent rebound even in BI3406-sensitive PDOs with no change of KRAS downstream effector genes, thus suggesting upregulation of guanine nucleotide exchange factor as potential cellular adaptation mechanisms to SOS1 inhibition. Taken together, our results show that high SOS1/SOS2 protein expression ratio predicts sensitivity to SOS1 inhibition and support further clinical development of SOS1-targeting agents in CRC.


Biogenesis, delivery, and function of extracellular RNA.
The Extracellular RNA (exRNA) Communication Consortium was launched by the National Institutes of Health to focus on the extent to which RNA might function in a non-cell-autonomous manner. With the availability of increasingly sensitive tools, small amounts of RNA can be detected in serum, plasma, and other bodily fluids. The exact mechanism(s) by which RNA can be secreted from cells and the mechanisms for the delivery and uptake by recipient cells remain to be determined. This review will summarize current knowledge about the biogenesis and delivery of exRNA and outline projects seeking to understand the functional impact of exRNA.


LEUNIG_HOMOLOG Mediates MYC2-Dependent Transcriptional Activation in Cooperation with the Coactivators HAC1 and MED25.
Groucho/Thymidine uptake 1 (Gro/Tup1) family proteins are evolutionarily conserved transcriptional coregulators in eukaryotic cells. Despite their prominent function in transcriptional repression, little is known about their role in transcriptional activation and the underlying mechanism. Here, we report that the plant Gro/Tup1 family protein LEUNIG_HOMOLOG (LUH) activates MYELOCYTOMATOSIS2 (MYC2)-directed transcription of JAZ2 and LOX2 via the Mediator complex coactivator and the histone acetyltransferase HAC1. We show that the Mediator subunit MED25 physically recruits LUH to MYC2 target promoters that then links MYC2 with HAC1-dependent acetylation of Lys-9 of histone H3 (H3K9ac) to activate JAZ2 and LOX2 Moreover, LUH promotes hormone-dependent enhancement of protein interactions between MYC2 and its coactivators MED25 and HAC1. Our results demonstrate that LUH interacts with MED25 and HAC1 through its distinct domains, thus imposing a selective advantage by acting as a scaffold for MYC2 activation. Therefore, the function of LUH in regulating jasmonate signaling is distinct from the function of TOPLESS, another member of the Gro/Tup1 family that represses MYC2-dependent gene expression in the resting stage.


Changing health policies and clinical laboratory services.
Recent changes in Medicare financing policy and in state certificate of need (CON) policies have greatly altered the environment in which hospitals and physicians must operate. These policy changes likely will influence the development, adoption, and use of new technologies, including those employed in clinical laboratories. Some hospitals, particularly large academic health centers, are responding to the new incentives under Medicare prospective payment by (1) implementing integrated case mix and financial information systems, (2) monitoring and evaluating physician practice patterns, (3) improving the quality of analyses underlying their technology acquisition decisions, and (4) striving toward improved relations with medical staff. Clinical laboratories are also responding to these financial pressures through (1) more cautious adoption of new technologies, (2) selective automation of tests that reduce direct costs, (3) more prudent "make-or-buy" decisions regarding test performance sites, and (4) focused effort to improve turnaround times for inpatient testing and reporting.


Efficiency considerations of donor fatigue, universal access to ARTs and health systems.
To investigate trends in official development assistance for health, HIV and non-HIV activities over time and to discuss the efficiency implications of these trends in the context of achieving universal access to treatment and health systems. Official development assistance for health, HIV programmes and non-HIV programmes were tracked using data from 2000 to 2009. A review of the literature on efficiency, treatment and health systems was conducted. The rate of growth of donor funding to HIV programmes has slowed in recent years at levels below those required to sustain programmes and to move towards universal access to treatment. These trends are likely due to increased pressure on foreign aid budgets and donor fatigue for HIV programmes. There is great need to consider how the limited resources available can be used most efficiently to increase the number of lives saved and to ensure that these resources also benefit health systems. Improving efficiency is much more than just improving the productive efficiency and also about ensuring that resources are going to where they will be the most beneficial and making investments that are the most efficient over time. These choices may be essential to achieving the goal of universal access to treatment as well as the sustainability of these programmes.


The gap in injury mortality rates between urban and rural residents of Hubei Province, China.
Injury is a growing public health concern in China. Injury death rates are often higher in rural areas than in urban areas in general. The objective of this study is to compare the injury mortality rates in urban and rural residents in Hubei Province in central China by age, sex and mechanism of injury. Using data from the Disease Surveillance Points (DSP) system maintained by the Hubei Province Centers for Disease Control and Prevention (CDC) from 2006 to 2008, injury deaths were classified according to the International Classification of Disease-10th Revision (ICD-10). Crude and age-adjusted annual mortality rates were calculated for rural and urban residents of Hubei Province. The crude and age-adjusted injury death rates were significantly higher for rural residents than for urban residents (crude rate ratio 1.9, 95% confidence interval 1.8-2.0; adjusted rate ratio 2.4, 95% confidence interval 2.3-2.4). The age-adjusted injury death rate for males was 81.6/100,000 in rural areas compared with 37.0/100,000 in urban areas; for females, the respective rates were 57.9/100,000 and 22.4/100,000. Death rates for suicide (32.4 per 100,000 vs 3.9 per 100,000), traffic-related injuries (15.8 per 100,000 vs 9.5 per 100,000), drowning (6.9 per 100,000 vs 2.3 per 100,000) and crushing injuries (2.0 per 100,000 vs 0.7 per 100,000) were significantly higher in rural areas. Overall injury death rates were much higher in persons over 65 years, with significantly higher rates in rural residents compared with urban residents for suicide (279.8 per 100,000 vs 10.7 per 100,000), traffic-related injuries, and drownings in this age group. Death rates for falls, poisoning, and suffocation were similar in the two geographic groups. Rates of suicide, traffic-related injury deaths and drownings are demonstrably higher in rural compared with urban locations and should be targeted for injury prevention activity. There is a need for injury prevention policies targeted at elderly residents, especially with regard to suicide prevention in rural areas in Central China.


Pharmacokinetics of 3,3',5,5'-tetrachlorobiphenyl in the male rat.
The pharmacokinetics of 3,3',5,5'-tetrachlorobiphenyl (4-CB) was studied to determine the importance of chlorine position to the kinetics of polychlorinated biphenyl (PCB) distribution, metabolism, and excretion by the rat. Tissue samples and excreta were collected at times ranging from 2 hr to 42 days after intravenous injection of a single 0.6-mg/kg dose of 14C-labeled 4-CB. The relative fractions of 4-CB and its metabolites were determined in selected samples, and 4-hr bile cannulations were performed to establish the rate of biliary excretion. A mathematical model description of PCB kinetics in the rat was defined here for 4-CB and used to generate tissue and excretion concentration-time profiles for both the parent compound and its metabolites. Model predictions were in good agreement with the experimental data, and showed how the individual pharmacokinetics of 4-CB compared to several other PBC's. The rate of 4-CB metabolism and excretion was slower than would be predicted according to its degree of chlorination. The relative rates of metabolism for 4-CB and five other differently chlorinated biphenyls could be ordered according to the number of adjacent unsubstituted carbon-atom pairs they contained. It appears that chlorine position is more important than degree of chlorination because the former can alter the availability of vicinal hydrogens thought to facilitate rapid PCB metabolism by mammals.


Reduction in phosphoenolpyruvate carboxykinase in rat liver parenchymal cells following experimentally induced cholestasis.
The effect of experimentally induced cholestasis on the amount of phosphoenolpyruvate carboxykinase (PEPCK) was studied immunohistochemically in rat liver parenchyma. In control liver, the enzyme was mainly localized periportally and, although the enzyme content was much reduced, this distribution pattern was maintained up to 2 weeks after ligation of the common bile duct. At 4 and 8 weeks after ligation the enzyme content in parenchymal cells remained low, but became distributed homogeneously throughout the liver parenchyma. This suggests that after bile duct ligation, gluconeogenesis from lactate is impaired. This may well be the cause of the adaptive changes to enhance the glycogenolytic capacity of parenchymal cells to maintain as far as possible a constant blood glucose level.


Non-alcoholic fatty liver disease is not independent risk factor for cardiovascular disease event: A cohort study.
There are no consistent results between previous studies for an independent association between non-alcoholic fatty liver disease (NAFLD) and cardiovascular disease (CVD) events. To determine if there is an independent association between NAFLD and CVD events. In the present study, valid outcome data of 4808 subjects were available for phase 2 of our cohort study. These subjects had been followed up for seven years from phase 1, beginning in 2009-2010 to phase 2 during 2016-2017. Simple and multiple Cox proportional models were used to determine the association between NAFLD in the primary phase of the cohort and subsequent fatal and non-fatal CVD events during follow-up. The incidence of non-fatal CVD events in males with NAFLD was significantly higher (P = 0.004) than in males without NAFLD. A positive association was demonstrated between NAFLD and non-fatal CVD events in males (Hazard ratio = 1.606; 95%CI: 1.166-2.212; P = 0.004) by the simple Cox proportional hazard model, but no independent association was detected between these in the multiple Cox models. No independent association was detected between NAFLD and CVD. It is likely that diabetes mellitus and age may be the principle mediators in this regard.


Robustness and accuracy of cell division in Escherichia coli in diverse cell shapes.
Cell division in typical rod-shaped bacteria such as Escherichia coli shows a remarkable plasticity in being able to adapt to a variety of irregular cell shapes. Here, we investigate the roles of the Min system and the nucleoid-occlusion factor SlmA in supporting this adaptation. We study "squeezed" E. coli in narrow nanofabricated channels where these bacteria exhibit highly irregular shapes and large volumes. Despite the severely anomalous morphologies we find that most of these bacteria maintain their ability to divide into two equally sized daughters with an accuracy comparable to that of normal rod-shaped cells (about 4%). Deletion of either slmA or minC shows that the molecular systems associated with these genes are largely dispensable for accurate cell division in these irregular cell shapes. Using fluorescence time-lapse microscopy, we determine that the functionality of the Min system is affected by the cell shape, whereas the localization of a nucleoid relative to the cell division proteins (the divisome) remains unperturbed in a broad spectrum of morphologies, consistent with nucleoid occlusion. The observed positioning of the nucleoid relative to the divisome appears not to be affected by the nucleoid-occlusion factor SlmA. The current study underscores the importance of nucleoid occlusion in positioning the divisome and shows that it is robust against shape irregularities.


Polyandrous females acquire indirect benefits in a nuptial feeding species.
The relative force of direct and indirect selection underlying the evolution of polyandry is contentious. When females acquire direct benefits during mating, indirect benefits are often considered negligible. Although direct benefits are likely to play a prominent role in the evolution of polyandry, post-mating selection for indirect benefits may subsequently evolve. We examined whether polyandrous females acquire indirect benefits and quantified direct and indirect effects of multiple mating on female fitness in a nuptial gift-giving spider (Pisaura mirabilis). In this system, the food item donated by males during mating predicts direct benefits of polyandry. We compared fecundity, fertility and survival of singly mated females to that of females mated three times with the same (monogamy) or different (polyandry) males in a two-factorial design where females were kept under high and low feeding conditions. Greater access to nutrients and sperm had surprisingly little positive effect on fitness, apart from shortening the time until oviposition. In contrast, polyandry increased female reproductive success by increasing the probability of oviposition, and egg hatching success indicating that indirect benefits arise from mating with several different mating partners rather than resources transferred by males. The evolution of polyandry in a male-resource-based mating system may result from exploitation of the female foraging motivation and that indirect genetic benefits are subsequently derived resulting from co-evolutionary post-mating processes to gain a reproductive advantage or to counter costs of mating. Importantly, indirect benefits may represent an additional explanation for the maintenance of polyandry.


MiR-499 regulates cell proliferation and apoptosis during late-stage cardiac differentiation via Sox6 and cyclin D1.
MiR-499 is a cardiac-abundant miRNA. However, the biological functions of miR-499 in differentiated cardiomyocytes or in the cardiomyocyte differentiation process is not very clear. Sox6 is believed to be one of its targets, and is also believed to play a role in cardiac differentiation. Therefore, our aim was to investigate the association between Sox6 and miR-499 during cardiac differentiation. Using a well-established in vitro cardiomyocyte differentiation system, mouse P19CL6 cells, we found that miR-499 was highly expressed in the late stage of cardiac differentiation. In cells stably transfected with miR-499 (P-499 cells), it was found that miR-499 could promote the differentiation into cardiomyocytes at the early stage of cardiac differentiation. Notably, cell viability assay, EdU incorporation assay, and cell cycle profile analysis all showed that the P-499 cells displayed the distinctive feature of hyperplastic growth. Further investigation confirmed that miR-499 could promote neonatal rat cardiomyocyte proliferation. MiR-499 knock-down enhanced apoptosis in the late differentiation stage in P19CL6 cells, but overexpression of miR-499 resulted in a decrease in the apoptosis rate. Sox6 was identified as a direct target of miR-499 and its expression was detected from day 8 or day 10 of cardiac differentiation of P19CL6 cells. Sox6 played a role in cell viability, inhibited cell proliferation and promoted cell apoptosis in P19CL6 cells and cardiomyocytes. The overexpression of Sox6 could reverse the proliferation and anti-apoptosis effects of miR-499. It was also found that miR-499 might exert its function by regulating cyclin D1 via its influence on Sox6. miR-499 probably regulates the proliferation and apoptosis of P19CL6 cells in the late stage of cardiac differentiation via its effects on Sox6 and cyclin D1.


ATP citrate-lyase and glycogen synthase kinase-3 beta in 3T3-L1 cells during differentiation into adipocytes.
ATP citrate-lyase (CL), acetyl-CoA carboxylase (ACC) and glycogen synthase kinase-3 beta (GSK-3 beta) levels were measured in cytosol from 3T3-L1 cells during differentiation from fibroblasts into fat-cells. Protein levels were estimated from immunoblots using specific antisera. Cytosol from confluent cells contain significant amounts of GSK-3 beta, which fell during differentiation of these cells into adipocytes. CL from confluent cells was found to be mostly in the form of a single protein band of apparent mass 110 kDa. Levels of CL and ACC increased during cell differentiation into adipocytes. During the first 3 days of differentiation, CL migration changed, and it was expressed as a complex of protein bands of apparent mass 110 kDa, 113 kDa and 115 kDa. At later stages of differentiation, when these cells had assumed the phenotype of fat-cells, they expressed CL mainly as protein bands of 110 and 113 kDa. When samples containing these bands were treated with alkaline phosphatase, the 113 kDa protein band collapsed into the 110 kDa species. This suggests that the slower-migrating species of CL is a higher-order phosphorylation state of the same protein. Furthermore, when purified CL, mostly expressed as the 110 kDa species, was phosphorylated with cyclic AMP-dependent protein kinase alone or together with GSK-3 and resolved by SDS/PAGE, the phosphorylated CL now migrated more slowly as the 113 kDa and 115 kDa forms. CL phosphorylation was hormone-regulated, since, in samples from fat-cells that had the complex two-band pattern, when cultured in medium without serum or hormones, CL migration reverted to a single band of 110 kDa, similar to confluent cells. Treatment of these 'down-regulated' cells with insulin rapidly induced substantial amounts of the 113 kDa species, with a concomitant decrease in the 110 kDa species.


Gut Microbiota and Cognitive Function Among Women Living with HIV.
Altered gut microbiota has been associated with cognitive dysfunction and Alzheimer's disease, but little is known among people living with HIV. To examine associations between gut microbiota and cognitive impairment among women with or without HIV. This is a cross-sectional study of 446 women (302 HIV+) who had completed a neuropsychological test battery and stool sample collected within 1 year. Gut microbiota composition was quantified using 16SV4 rRNA gene sequencing and microbial functional pathways were predicted using PICRUSt. Cognitive domains included attention, executive function, learning, memory, fluency, processing speed, and motor function. Cognitive impairment was defined as two or more domains with T scores < 1 SD below mean. ANCOM-II was used to identify taxa and functional pathways associated with cognitive impairment, and the associations were further examined by multivariable logistic regression. In overall sample, adjusting for multiple covariates including HIV status, we found that higher abundance of Methanobrevibacter, Odoribacter, Pyramidobacter, Eubacterium, Ruminococcus, and Gemmiger, and lower abundance of Veillonella were associated with cognitive impairment. The associations between these taxa and cognitive impairment were more profound in HIV+ women compared to HIV- women. Most associations with bacterial taxa were observed for learning and memory. We found accompanying microbial functional differences associated with cognitive impairment, including twelve enriched pathways and three depleted pathways. In women with or without HIV infection, this study identified multiple altered gut bacterial taxa and functional pathways associated with cognitive impairment, supporting the potential role of gut microbiota in cognitive dysfunction and Alzheimer's disease.


Bilateral versus unilateral placement of metal stents for inoperable high-grade malignant hilar biliary strictures: a multicenter, prospective, randomized study (with video).
The efficacy of palliative biliary drainage by using bilateral or unilateral self-expandable metal stents (SEMSs) for a malignant hilar biliary stricture (MHS) remains controversial. This prospective, randomized, multicenter study investigated whether bilateral drainage by using SEMSs is superior to unilateral drainage in patients with inoperable MHSs. Patients with inoperable high-grade MHSs who underwent palliative endoscopic insertion of bilateral or unilateral SEMSs were enrolled. The main outcome measurements were the rate of primary reintervention for malfunction after successful placement of SEMSs, stent patency, technical and clinical success rates, adverse events, and survival duration. A total of 133 pathology-diagnosed patients were randomized to the bilateral group (n = 67) or the unilateral group (n = 66). The primary technical success rates were 95.5% (64/67) and 100% (66/66) in the bilateral and unilateral groups, respectively (P = .244). The clinical success rates were 95.3% (61/64) and 84.9% (56/66), respectively (P = .047). The primary reintervention rates based on the per-protocol analysis were 42.6% (26/61) in the bilateral group and 60.3% (38/63) in the unilateral group (P = .049). The median cumulative stent patency duration was 252 days in the bilateral group and 139 days in the unilateral group. The risk of stent patency failure was significantly higher in the unilateral group (log-rank test; P < .01). In a multivariate Cox proportional hazard model to assess stent patency, bilateral SEMS placement was a favorable factor (adjusted hazard ratio 0.30, 95% confidence interval, 0.172-0.521; P < .001). Survival probability and late adverse events were not different between the 2 groups. Unilateral and bilateral drainage strategies by using SEMSs had similar technical success rates, but bilateral drainage resulted in fewer reinterventions and more durable stent patency in patients with inoperable high-grade MHSs. (Clinical trial registration number: NCT02166970.).


Fetal swallowing: correlation of electromyography and esophageal fluid flow.
Patterns of fetal drinking behavior were quantified in five singleton fetuses (128 +/- 1 days gestation) by computer analysis of laryngeal-esophageal electromyography (EMG) and thoracic esophageal fluid flow. Esophageal flow was noted to be bidirectional with an average antegrade flow-to-retrograde flow ratio of 4.4 +/- 1.5. Retrograde esophageal flow coincided with diaphragmatic contractions, suggesting that inspiratory gastroesophageal pressure gradients may be operative. The regurgitated fluid was returned to the fetal rumen by thoracic esophageal contractions. Significant net fluid intake was noted during periods of primarily unidirectional antegrade esophageal flow. Fetuses swallowed an average of 35 +/- 9 ml/h during the 12-h study, extrapolating to an average daily volume of 840 +/- 224 ml. An EMG "propagated swallow" representing coordinated contractions of the thyrohyoid, nuchal esophagus, and thoracic esophagus averaged 43 +/- 3 swallows/h and was highly correlated with net esophageal flow. Nearly 60% of the total fluid intake occurred during "bouts" of propagated swallows, which represented only 8% of the study period. Bouts averaged 11.7 +/- 0.4 propagated swallows, lasted 2.1 +/- 0.2 min, and occurred every 27.6 +/- 1.7 min. Although EMG swallowing activity was similar, there were significant differences among fetuses in net esophageal fluid flow and volume per propagated swallow. We speculate that fetal fluid intake is dependent on the availability and physical properties of the swallowed fluid, whereas swallowing activity is primarily related to cortical maturation and thirst stimulation.


Hypertension evaluated in the public and private Brazilian health system hypertension in public and private service.
Hypertension (HT) remains the leading cause of death worldwide. In Brazil it is estimated that 35% of the adult population has HT and that about 20% of these have blood pressure values within the targets recommended for the reduction of cardiovascular risk. There are some data that point to different control rates in patients treated by cardiologists in public and private referral center and this is an important point to be investigated and discussed. To compare sociodemographic characteristics, body mass index (BMI), antihypertensive (AH) drugs, blood pressure (BP) and control rate in public (PURC) and private (PRRC) referral centers. A cross-sectional multicenter study that analyzed data from hypertensive patients assisted by the PURC (one in Midwest Region and other in Northeast region) and PRRC (same distribution). Variables analyzed: sex, age, BMI, classes, number of AH used and mean values of systolic and diastolic BP by office measurement and home blood pressure measurement (HBPM). Uncontrolled hypertension (HT) phenotypes and BP control rates were assessed. Descriptive statistics and χ2 tests or unpaired t-tests were performed. A significance level of p < 0.05 was considered. A predominantly female (58.9%) sample of 2.956 patients and a higher prevalence of obesity in PURC (p < 0.001) and overweight in PRRC (p < 0.001). The mean AH used was 2.9 ± 1.5 for PURC and 1.4 ± 0.7 for PRRC (p < 0.001). Mean systolic and diastolic BP values were higher in PURC as were rates of uncontrolled HT of 67.8% and 47.6% (p < 0.001) by office measurement and 60.4% and 35.3% (p < 0.001) by HBPM in PURC and PRRC, respectively. Patients with HT had a higher prevalence of obesity in the PURC and used almost twice as many AH drugs. BP control rates are worse in the PURC, on average 15.3 mmHg and 12.1 mmHg higher than in the PRRC by office measurement.


Randomized, dim light controlled, crossover test of morning bright light therapy for rest-activity rhythm disorders in patients with vascular dementia and dementia of Alzheimer's type.
The authors compared the therapeutic effect of morning bright and dim light exposure on rest-activity (R-A) rhythm disorders in patients with vascular dementia (VD) and patients with dementia of Alzheimer's type (DAT). Participants in this study were 12 patients with VD (M/F = 5/7; average age = 81 years) and 10 patients with DAT (M/F = 4/6; average age = 78 years). They were exposed to 2 weeks of bright light (BL; 5000-8000 lux) and 2 weeks of dim light (DL; 300 lux) in the morning (09:00-11:00) in a randomized crossover design in which the 2-week treatment period took place between pretreatment (1 week) and posttreatment (1 week) periods. Continuous R-A monitoring was performed at 1-minute intervals throughout the study using an actigraph around the nondominant wrist. The BL exposure for 2 weeks induced a significant reduction in both nighttime activity and percentages of nighttime activity to total activity compared with the pretreatment period, as well as compared with the DL condition in the VD group, but not in the DAT group. These findings support the assumption that the therapeutic efficacies of morning BL exposure are prominent in VD patients and are mainly due to its photic effect rather than nonphotic effects such as the intensification of social interaction accompanying light therapy.


Responses of model cortical neurons to temporal interference stimulation and related transcranial alternating current stimulation modalities.
Objective.Temporal interference stimulation (TIS) was proposed as a non-invasive, focal, and steerable deep brain stimulation method. However, the mechanisms underlying experimentally-observed suprathreshold TIS effects are unknown, and prior simulation studies had limitations in the representations of the TIS electric field (E-field) and cerebral neurons. We examined the E-field and neural response characteristics for TIS and related transcranial alternating current stimulation modalities.Approach.Using the uniform-field approximation, we simulated a range of stimulation parameters in biophysically realistic model cortical neurons, including different orientations, frequencies, amplitude ratios, amplitude modulation, and phase difference of the E-fields, and obtained thresholds for both activation and conduction block.Main results. For two E-fields with similar amplitudes (representative of E-field distributions at the target region), TIS generated an amplitude-modulated (AM) total E-field. Due to the phase difference of the individual E-fields, the total TIS E-field vector also exhibited rotation where the orientations of the two E-fields were not aligned (generally also at the target region). TIS activation thresholds (75-230 V m-1) were similar to those of high-frequency stimulation with or without modulation and/or rotation. For E-field dominated by the high-frequency carrier and with minimal amplitude modulation and/or rotation (typically outside the target region), TIS was less effective at activation and more effective at block. Unlike AM high-frequency stimulation, TIS generated conduction block with some orientations and amplitude ratios of individual E-fields at very high amplitudes of the total E-field (>1700 V m-1).Significance. The complex 3D properties of the TIS E-fields should be accounted for in computational and experimental studies. The mechanisms of suprathreshold cortical TIS appear to involve neural activity block and periodic activation or onset response, consistent with computational studies of peripheral axons. These phenomena occur at E-field strengths too high to be delivered tolerably through scalp electrodes and may inhibit endogenous activity in off-target regions, suggesting limited significance of suprathreshold TIS.


A method for the determination of estrogen receptor concentration in calf uterus and other tissues using an aqueous two-phase system.
An assay for the cytoplasmic estrogen receptor in calf, human and rat uterus has been developed. The method is based on partial separation of free and bound estradiol (E2) by means of an aqueous two-phase system containing dextran and poly(ethylene glycol), respectively, in the two phases. Low-speed supernatant from uterus homogenate is equilibrated with E2 and [3H]E2. A two-phase mixture is then added and bound E2 will partition into the lower phase while free E2 is distributed in both phases according to its partition coefficient. The amounts of bound and free E2 are calculated and the receptor concentration and association constant are obtained from a Scatchard plot. No dissociation of bound E2 in the phase system could be demonstrated at 4 degrees C. The interassay coefficient of variation for receptor concentration at 4 degrees C was 20 and 14% for calf and human uterus, respectively. The intraassay variation for receptor concentration in calf uterus determined at 4 degrees C and 23 degrees C was 7.1 and 4.1%, respectively. The influence of freezing the tissue and supernatant preparation was examined and results from supernatant preparations obtained with different centrifugations were compared. The method is simple and rapid, permitting large numbers of samples to be handled efficiently by a single technician.


Abdominal hernia with formation of a urate concretion in a cockatiel.
A female cockatiel had a 2-week history of abdominal distention, lethargy, and diarrhea. The cockatiel had a history of frequent egg-laying, and the owner suspected that she was egg-bound. A solid mass was removed through the cloaca and found to be a concretion of urates that had formed within a hernial pouch of the caudal abdominal musculature. Diagnosis was aided by contrast radiography, and surgery was performed. However, the abdominal hernias recurred 16 months later. The continual egg-laying probably predisposed this bird to hernia formation. Abdominal hernias in birds may be a consequence of continual egg-laying and associated hormonal effects leading to a weakening of abdominal musculature.


Drugs in context: a historical perspective on theories of psychopharmaceutical efficacy.
This article demonstrates that psychoanalysis and socially oriented psychiatry were crucial to the understanding and adoption of the first effective psychopharmaceuticals in North American psychiatry. In the 1950s and the early 1960s, psychoanalysts, socially oriented psychiatrists, and biologists collaborated, debated, and organized interdisciplinary conferences to situate the biochemistry of new psychopharmaceuticals, such as chlorpromazine, in the broader psychosocial context of patients' lives. Psychoanalytical and sociological perspectives not only helped American psychiatrists explain the mechanism of drug action in research but also established the professional authority of psychiatrists over the new pharmaceuticals. As modern pharmacology narrows its focus to microscopic targets in the body, I argue that this early drug research illustrates the present-day need for holistic and interdisciplinary approaches to drug response that acknowledge the psychosocial significance of psychiatric medication in the lives of individuals.


Longitudinal study of adaptation to the stress of bone marrow transplantation.
This prospective longitudinal study of adaptation to bone marrow transplantation (BMT) addressed three questions: (1) When during BMT do individuals experience the greatest distress? (2) What factors are associated with this distress? (3) Are there variables that could be potential clinical indicators of persons in greatest need of preventive intervention? One hundred one participants undergoing either an autologous or allogeneic BMT completed questionnaires before hospitalization, before bone marrow infusion, 7 days and 14 days after transplantation, and then 1 month, 3 months, and 12 months after hospitalization. Adaptation was indicated by the degree of emotional distress. Independent variables were personal control, social support from specific sources, cognitive response, self-perception, and coping strategies, controlling for symptomatology. The greatest emotional distress occurred after admission to the hospital and before the bone marrow infusion. Anxiety and depression decreased 1 week after the transplant, although symptomatology increased during this time. The periods of least emotional distress were 3 months and 1 year after transplantation. Factors that accounted for the greatest variance in emotional distress/adaptation were the degree of emotional distress at baseline, personal control, cognitive response, and symptomatology. According to this longitudinal study, which includes pretransplant data, data from in-hospital transplantation, and posttransplant data, (1) psychosocial vulnerability of these BMT recipients was greatest during hospitalization before the transplant, (2) perceived personal control may be a potential indicator of vulnerability to secondary psychosocial morbidity, and (3) the demonstrated significance of psychosocial well-being before BMT indicates the importance of obtaining prospective data for both research and clinical purposes.


The role of community outreach and engagement in evaluation of NCI Cancer Center Support Grants.
National Cancer Institute (NCI)-designated cancer centers are required to consider their impact on the catchment area they serve. These activities are facilitated by community outreach and engagement (COE) activities as specified in the Cancer Center Support Grant (CCSG) request for applications. While the critical importance of COE activities to NCI-designated cancer centers is well known, it is less clear what impact the COE component has on the overall CCSG merit descriptor and score. We undertook an online survey of all 62 NCI-designated Comprehensive and Clinical centers who reported their COE merit descriptor and overall CCSG priority score as of Fall 2021. Of 48 (77%) of responding centers, we identified a strong correlation between the COE merit descriptor and the overall numerical CCSG score received by the center (Spearman's rank correlation coefficient r = 0.360, p = 0.0053). When stratifying this relationship by center type, we observed a very strong correlation between COE and CCSG ratings for comprehensive cancer centers (n = 40; r = 0.544; p = 0.0003) but not for non-comprehensive cancer centers (n = 8; r = 0.073; p = 0.864). COE component merit descriptors for comprehensive cancer center CCSG evaluations are strongly correlated with the overall cancer center review score.


Rat brain N-methyl-D-aspartate receptors require multiple molecules of agonist for activation.
N-Methyl-D-aspartate (NMDA) receptors mediate important physiological and pathological processes, including long term potentiation and neuronal excitotoxicity. Elucidation of mechanisms underlying NMDA receptor functioning will promote understanding of the molecular bases of NMDA receptor-mediated processes. The localization of the phencyclidine (PCP) receptor within the ionophore of the NMDA receptor-gated ion channel permits the binding of PCP receptor ligands to serve as a functional marker of channel activation. We have previously demonstrated that the highly selective PCP receptor ligand [3H]MK-801 displays multiexponential kinetics of association, indicating that the NMDA receptor functions according to a multistate model. Using the fast component of [3H]MK-801 binding to PCP receptors as a marker for activated NMDA channels, we demonstrate here a Hill coefficient of 2 for activation of NMDA channels by L-glutamate. A multistate model of NMDA receptor functioning analogous to the model known to account for the functioning of nicotinic cholinergic and gamma-aminobutyric acidA receptors fits well to our experimental data, supporting the concept that the NMDA receptor is properly classified in the Class 1 superfamily of ligand-gated channels.


A helical conotoxin from Conus imperialis has a novel cysteine framework and defines a new superfamily.
Cone snail venoms are a rich source of peptides, many of which are potent and selective modulators of ion channels and receptors. Here we report the isolation and characterization of two novel conotoxins from the venom of Conus imperialis. These two toxins contain a novel cysteine framework, C-C-C-CC-C, which has not been found in other conotoxins described to date. We name it framework XXIII and designate the two toxins im23a and im23b; cDNAs of these toxins exhibit a novel signal peptide sequence, which defines a new K-superfamily. The disulfide connectivity of im23a has been mapped by chemical mapping of partially reduced intermediates and by NMR structure calculations, both of which establish a I-II, III-IV, V-VI pattern of disulfide bridges. This pattern was also confirmed by synthesis of im23a with orthogonal protection of individual cysteine residues. The solution structure of im23a reveals that im23a adopts a novel helical hairpin fold. A cluster of acidic residues on the surface of the molecule is able to bind calcium. The biological activity of the native and recombinant peptides was tested by injection into mice intracranially and intravenously to assess the effects on the central and peripheral nervous systems, respectively. Intracranial injection of im23a or im23b into mice induced excitatory symptoms; however, the biological target of these new toxins has yet to be identified.


[Hematomas of the lenticular nucleus and pons in a hypertensive patient].
Secondary pontine hemorrhages are not rare in intracranial hypertension caused by a supratentorial mass lesion. Cerebral herniation and caudal dislocation of the brainstem account for the secondary pontine hemorrhages. In the present case, two unrelated hemorrhages were demonstrated by an early CT. Neurological findings and CT thus suggest that this is a rare case in which supratentorial and infratentorial hypertensive intracerebral hemorrhages occurred simultaneously.


Hippocampal, but not amygdala, activity at encoding correlates with long-term, free recall of nonemotional information.
Participation of two medial temporal lobe structures, the hippocampal region and the amygdala, in long-term declarative memory encoding was examined by using positron emission tomography of regional cerebral glucose. Positron emission tomography scanning was performed in eight healthy subjects listening passively to a repeated sequence of unrelated words. Memory for the words was assessed 24 hr later with an incidental free recall test. The percentage of words freely recalled then was correlated with glucose activity during encoding. The results revealed a striking correlation (r = 0.91, P < 0.001) between activity of the left hippocampal region (centered on the dorsal parahippocampal gyrus) and word recall. No correlation was found between activity of either the left or right amygdala and recall. The findings provide evidence for hippocampal involvement in long-term declarative memory encoding and for the view that the amygdala is not involved with declarative memory formation for nonemotional material.


In vitro concentrative accumulation of D-xylose by jejunum from horses and rabbits.
Accumulation of D-xylose by jejunal mucosa from healthy horses and rabbits was studied in vitro. When tissue sheets were incubated with 1 mM D-xylose for 60 minutes, mucosa from horses and rabbits accumulated D-xylose against a concentration gradient. There was no accumulation when equine specimens were incubated with 5 mM D-xylose. By comparison, equine jejunum accumulated D-glucose against a concentration gradient when incubated in 5 mM D-glucose. In equine and rabbit jejunum, 13.3 +/- 7.0% and 36 +/- 11.0%, respectively, of accumulated D-xylose was phosphorylated when sheets were incubated in 1 mM D-xylose. Short-circuit current and potential difference were lower in equine jejunum than in rabbit jejunum, possibly because of differences in tissue thickness. None of the transmucosal electrical measurements increased after addition of D-xylose (1 mM and 5 mM) or D-glucose (5 mM). The active transport system for D-xylose has a low affinity for this sugar and becomes saturated at low intraluminal concentrations. Therefore, abnormal D-xylose absorption test results in horses are more likely caused by abnormalities in mucosal surface area and mucosal permeability than by abnormalities of nutrient carbohydrate absorption.


[Nirmatrelvir/ritonavir drug-drug interactions in primary care outpatients].
The oral antiviral nirmatrelvir/ritonavir interacts with a range of drugs. Candidate patients to receive this antiviral agent are usually vulnerable, multipathological and polymedicated. The objective is to evaluate the pharmaceutical validation prior to the administration of the antiviral. Nirmatrelvir/ritonavir drug-drug interactions to patients' usual treatment medications were checked in product information and in the UpToDate® and the University of Liverpool® interaction tools. We included validated prescriptions between April/2022 and April/2023 by a Primary Care pharmacist. Of the 159 study patients, 168 interactions were found in 83 individuals, which may have led to changes of their usual treatment. Statins (25.6%), anticoagulants (10.7%), and antihypertensives (10.7%) were the most frequently implicated therapeutic groups. Discontinuation (53.0%) and dose reduction (22.6%) were the most common treatment changes. Our search of potential drug interactions and subsequent dose adjustments and modifications of the patient's usual treatment has helped avoid potential toxicities ensuring a safe use of nirmatrelvir/ritonavir.


Theoretical studies on the photoinduced rearrangement mechanism of α-santonin.
α-Santonin is the first organic compound observed to feature a photoinduced rearrangement and is now known to undergo a series of photochemical processes under UV irradiation. On the basis of the considerable interest of this system as a prototype, and of the yet limited insights reached for the basic photo mechanisms, we calculate the high-level electronic structures and explore the potential energy surfaces (PES) of α-santonin in the ground and lowest-lying excited states, their couplings, and the possible photoinduced isomerization pathways. The calculations identify the low-lying singlet excited state (1)(nπ*) accessible under light irradiation, which decays to the low-energy (3)(ππ*) state through an intersystem crossing in the Franck-Condon region to initiate the photoinduced rearrangement. The initial reaction from the C3-C5 bond coupling, which takes place on the (3)(ππ*) state potential energy surface, leads to a three-membered alkyl-ring compound intermediate state INT. The following photochemical reactions have the possibility to arise from two distinct C-C bond cleavages, C4-C5 and C3-C4, denoted as path A and path B. Path A is favored both dynamically on the excited-state PES and thermodynamically on the ground-state PES in vacuo. Experiments show that it also becomes the dominant photoinduced rearrangement process in the crystal, which can be explained by considering the requirement for less space and the stacking effect under the confined environment. Path B is dynamical advantaged both on the ground- and excited-state PESs in a weak polar solvent, such as dioxane. Once the biradical intermediate B-INT is accessible on the ground-state PES, the formation of the product B-P is almost barrier free.


Antitumor promoting potential of selected phytochemicals derived from spices: a review.
Carcinogenesis is a multistep process exhibiting deregulation in multiple cellular signaling pathways. Therefore, specific agent based treatments that target only one pathway usually fail in cancer therapy. The combination treatments using chemotherapeutic agents with distinct molecular mechanisms are considered more promising for higher efficacy; however, using multiple agents contributes to added toxicity. However, the in-vitro and in-vivo studies in the last few decades have demonstrated that some phytochemicals derived from 'natural products' such as fruits, vegetables and certain spices, referred to as chemopreventive agents, including capsaicin, trans-anethole, thymoquinone, diosgenin, allicin, can not only reduce the risk of acquiring specific cancer but also have been shown to suppress cancer cell proliferation, inhibit growth factor signaling pathways, induce apoptosis, inhibit nuclear factor-κB, AP-1, Akt, MAPK, Wnt, Notch, p53, AR, ER, and JAK-STAT, etc., activation pathways, inhibit angiogenesis, suppress the expression of antiapoptotic proteins, and inhibit cyclooxygenase-2. This study describes the above active components of some of the major spices, their mechanisms of action and their potential in prevention of various cancers.


Attitude towards reciprocity as a motive for oocyte donation.
Finding out whether patients would be motivated by reciprocity when considering donating oocytes to others. This is a prospective monocentric study in the CECOS of the Centre Hospitalier Universitaire (CHU) of Rennes (France) on the opinion of patients regarding reciprocity. Couples who had a child with donor sperm were asked whether they would consider oocyte donation as a way of giving something back. Twenty six couples and one man answered the questionnaire. About half of the respondents (49%) felt that they should contribute to the system from which they benefitted. Although the patients would benefit from a reduction in waiting time, this advantage was only important for one in four persons. The only items on which the answers between men and women were significantly different concerned the results of the donation: women would think more often about the potential recipient and the child and they more often wanted to know whether children were born from their donation. The results show that beside altruism, reciprocity may be an important moral reason for people to donate gametes.


Estimating fluctuating selection in age-structured populations.
In age-structured populations, viability and fecundity selection of varying strength may occur in different age classes. On the basis of an original idea by Fisher of weighting individuals by their reproductive value, we show that the combined effect of selection on traits at different ages acts through the individual reproductive value defined as the stochastic contribution of an individual to the total reproductive value of the population the following year. The selection differential is a weighted sum of age-specific differentials that are the covariances between the phenotype and the age-specific relative fitness defined by the individual reproductive value. This enables estimation of weak selection on a multivariate quantitative character in populations with no density regulation by combinations of age-specific linear regressions of individual reproductive values on the traits. Demographic stochasticity produces random variation in fitness components in finite samples of individuals and affects the statistical inference of the temporal average directional selection as well as the magnitude of fluctuating selection. Uncertainties in parameter estimates and test power depend strongly on the demographic stochasticity. Large demographic variance results in large uncertainties in yearly estimates of selection that complicates detection of significant fluctuating selection. The method is illustrated by an analysis of age-specific selection in house sparrows on a fitness-related two-dimensional morphological trait, tarsus length and body mass of fledglings.


Improvement Initiative to Develop and Implement a Tool for Detecting Drug-Drug Interactions During Oncology Clinical Trial Enrollment Eligibility Screening.
Screening subjects for drug-drug interactions (DDIs) before enrollment in oncology clinical trials is integral to ensuring safety, but standard procedures or tools are not readily available to screen DDI in this setting. Our objectives were to develop a DDI screening tool for use during oncology clinical trial enrollment and to test usability in single-center and multicenter pilot studies. A multistage approach was used for this quality improvement intervention. Semistructured interviews with individuals responsible for DDI screening were conducted to develop a prototype tool. The tool was used for screening DDI in subjects enrolling in National Clinical Trials Network trials of commercially available agents during a single-center 3-month pilot. Improvements were made, and a 3-month multicenter pilot was conducted at volunteer SWOG Cancer Research Network sites. Participants were surveyed to determine tool usability and efficiency. A tool was developed from semistructured interviews. A critical feature was reporting which medications had specific pharmacokinetic and pharmacodynamic characteristics including transporter and cytochrome P450 substrates, inhibitors, or inducers and QT prolongation. In the 12-site study, average (SD) DDI screening time for each patient decreased by 15.7 (10.2) minutes (range, 3-35 minutes; P < 0.001). Users reported the tool highly usable, with >90% agreeing with all positive usability characterizations and disagreeing with all negative complexity characterizations. A DDI screening tool for oncology clinical trial enrollment was created and its usability confirmed. Further testing with more diverse investigator sites and study drugs during eligibility screening is warranted to improve safety and data accuracy within clinical trials.


Modelling the potential health and economic benefits of reducing population sitting time in Australia.
Strong evidence indicates that excessive time spent sitting (sedentary behaviour) is detrimentally associated with multiple chronic diseases. Sedentary behaviour is prevalent among adults in Australia and has increased during the COVID-19 pandemic. Estimating the potential health benefits and healthcare cost saving associated with reductions in population sitting time could be useful for the development of public health initiatives. A sedentary behaviour model was developed and incorporated into an existing proportional, multi-state, life table Markov model (ACE-Obesity Policy model). This model simulates the 2019 Australian population (age 18 years and above) and estimates the incidence, prevalence and mortality of five diseases associated with sedentary behaviour (type 2 diabetes, stroke, endometrial, breast and colorectal cancer). Key model inputs included population sitting time estimates from the Australian National Health Survey 2014-2015, healthcare cost data from the Australian Institute of Health and Welfare (2015) and relative risk estimates assessed by conducting literature reviews and meta-analyses. Scenario analyses estimated the potential change in disease incidence as a result of changes in population sitting time. This, in turn, resulted in estimated improvements in long term health outcomes (Health-adjusted life years (HALYs)) and healthcare cost-savings. According to the model, if all Australian adults sat no more than 4 h per day, the total HALYs gained would be approximately 17,211 with health care cost savings of approximately A$185 million over one year. Under a more feasible scenario, where sitting time was reduced in adults who sit 4 or more hours per day by approximately 36 min per person per day (based on the results of the Stand Up Victoria randomised controlled trial), potential HALYs gained were estimated to be 3,670 and healthcare cost saving could reach A$39 million over one year. Excessive sedentary time results in considerable population health burden in Australia. This paper describes the development of the first Australian sedentary behaviour model that can be used to predict the long term consequences of interventions targeted at reducing sedentary behaviour through reductions in sitting time. These estimates may be used by decision makers when prioritising healthcare resources and investing in preventative public health initiatives.


'How did you cope with such concerns?': insights from a monitored online infertility peer support forum.
The Research Question posed was 'What coping strategies do fertility patients and trained peer supporters discuss in an online infertility peer support forum?' A thematic analysis was used to examine 244 online posts from a sample of 39 users (37 women and two men) for themes in coping with fertility-related stressors. Data were collected from Connect, a monitored online discussion forum with trained peer supporters. Connect users ranged from 27 to 44 years of age (mean = 34.38) and 33 (84.6%) were nulliparous at the time of the study. A variety of cognitive-reappraisal and practical management coping strategies was discussed across four themes: (i) balancing interpersonal relationships; (ii) partner support; (iii) uncertainty and lack of control; and (iv) positivity and negativity. Experiences of uncertainty and lack of control related to time and schedule, outcomes and waiting for physical symptoms, and the clinic. Connect users commonly discussed the helpfulness of coping strategies. Connect users actively requested experiential information about ways of coping from other patients and peer supporters, highlighting the importance of lived experience to those currently in treatment. Findings support conceptualizations of infertility coping as a process that is unique to the infertility treatment context and that may change throughout a patient's treatment. Trained peer supporters may benefit fertility patients by normalizing, reappraising, and providing practical strategies to ameliorate difficult infertility-related challenges.


Genetic studies of human neuropathic pain conditions: a review.
Numerous studies have shown associations between genetic variants and neuropathic pain disorders. Rare monogenic disorders are caused by mutations of substantial effect size in a single gene, whereas common disorders are likely to have a contribution from multiple genetic variants of mild effect size, representing different biological pathways. In this review, we survey the reported genetic contributors to neuropathic pain and submit them for validation in a 150,000-participant sample of the U.K. Biobank cohort. Successfully replicated association with a neuropathic pain construct for 2 variants in IL10 underscores the importance of neuroimmune interactions, whereas genome-wide significant association with low back pain (P = 1.3e-8) and false discovery rate 5% significant associations with hip, knee, and neck pain for variant rs7734804 upstream of the MAT2B gene provide evidence of shared contributing mechanisms to overlapping pain conditions at the molecular genetic level.


[Age-related differences and changes in clinical aspects and follow-up of cervix cancer since 1973].
In an attempt to clarify, whether the malignant potential of cervical cancer is age-dependent and whether this has changed in the last decade, we analysed the data from 1496 patients suffering from cervical cancer. The patients were treated between 1973 and 1988 at the second Universitäts-Frauenklinik, Lindwurmstrasse and at the Frauenklinik Grosshadern, both of the Ludwig-Maximilians-Universität München. After 1973, the total number of patients with cervical cancer decreased, whilst at the same time, the percentage of young patients increased. In the sub-group of surgically treated patients (n = 555), the percentage of patients with lymph node metastases increased after 1973. The increase in lymph node involvement was most pronounced in young patients with small tumours (stage Ib disease, 35 years of age). In this group, the percentage of lymph node involvement rose from 6.3% in 1973 to 38.6% in 1988. Lower stage disease was significantly more frequent in the young patients. Accordingly, the overall mean survival time was longer in the young patients. In the sub-group of surgically treated patients, however, when comparing equal stages, the survival time was shorter in the young patients. In small tumours (confined to the uterine cervix), pelvic lymph node involvement was more frequently seen in the younger patients. Secondary metastases developed more rapidly and significantly more often in the young patients. Our data supply evidence to indicate an enhanced malignancy of cervical cancer in young patients.


MR-Guided Kernel EM Reconstruction for Reduced Dose PET Imaging.
PET image reconstruction is highly susceptible to the impact of Poisson noise, and if shorter acquisition times or reduced injected doses are used, the noisy PET data become even more limiting. The recent development of kernel expectation maximisation (KEM) is a simple way to reduce noise in PET images, and we show in this work that impressive dose reduction can be achieved when the kernel method is used with MR-derived kernels. The kernel method is shown to surpass maximum likelihood expectation maximisation (MLEM) for the reconstruction of low-count datasets (corresponding to those obtained at reduced injected doses) producing visibly clearer reconstructions for unsmoothed and smoothed images, at all count levels. The kernel EM reconstruction of 10% of the data had comparable whole brain voxel-level error measures to the MLEM reconstruction of 100% of the data (for simulated data, at 100 iterations). For regional metrics, the kernel method at reduced dose levels attained a reduced coefficient of variation and more accurate mean values compared to MLEM. However, the advances provided by the kernel method are at the expense of possible over-smoothing of features unique to the PET data. Further assessment on clinical data is required to determine the level of dose reduction that can be routinely achieved using the kernel method, whilst maintaining the diagnostic utility of the scan.


Behaviors, movements, and transmission of droplet-mediated respiratory diseases during transcontinental airline flights.
With over 3 billion airline passengers annually, the inflight transmission of infectious diseases is an important global health concern. Over a dozen cases of inflight transmission of serious infections have been documented, and air travel can serve as a conduit for the rapid spread of newly emerging infections and pandemics. Despite sensational media stories and anecdotes, the risks of transmission of respiratory viruses in an airplane cabin are unknown. Movements of passengers and crew may facilitate disease transmission. On 10 transcontinental US flights, we chronicled behaviors and movements of individuals in the economy cabin on single-aisle aircraft. We simulated transmission during flight based on these data. Our results indicate there is low probability of direct transmission to passengers not seated in close proximity to an infectious passenger. This data-driven, dynamic network transmission model of droplet-mediated respiratory disease is unique. To measure the true pathogen burden, our team collected 229 environmental samples during the flights. Although eight flights were during Influenza season, all qPCR assays for 18 common respiratory viruses were negative.


Does maternal environmental tobacco smoke interact with social-demographics and environmental factors on congenital heart defects?
Congenital heart defects (CHDs) are a major cause of death in infancy and childhood. Major risk factors for most CHDs, particularly those resulting from the combination of environmental exposures with social determinants and behaviors, are still unknown. This study evaluated the main effect of maternal environmental tobacco smoke (ETS), and its interaction with social-demographics and environmental factors on CHDs in China. A population-based, matched case-control study of 9452 live-born infants and stillborn fetuses was conducted using the Guangdong Registry of Congenital Heart Disease data (2004-2014). The CHDs were evaluated by obstetrician, pediatrician, or cardiologist, and confirmed by cardia tomography/catheterization. Controls were randomly chosen from singleton newborns without any malformation, born in the same hospital as the cases and 1:1 matched by infant sex, time of conception, and parental residence (same city and town to ensure sufficient geographical distribution for analyses). Face-to-face interviews were conducted to collect information on demographics, behavior patterns, maternal disease/medication, and environmental exposures. Conditional logistic regression was used to estimate odds ratios and 95% confidence intervals of ETS exposure on CHDs while controlling for all risk factors. Interactive effects were evaluated using a multivariate delta method for maternal demographics, behavior, and environmental exposures on the ETS-CHD relationship. Mothers exposed to ETS during the first trimester of pregnancy were more likely to have infants with CHD than mothers who did not (aOR = 1.44, 95% CI 1.25-1.66). We also observed a significant dose-response relationship when mothers were exposed to ETS and an increasing number of risk factors and CHDs. There were greater than additive interactions for maternal ETS and migrant status, low household income and paternal alcohol consumption on CHDs. Maternal low education also modified the ETS-CHD association on the multiplicative scale. These findings may help to identify high-risk populations for CHD, providing an opportunity for targeted preventive interventions.


Promotion of tumor progression induced by continuous low-dose administration of antineoplastic agent gemcitabine or gemcitabine combined with cisplatin.
This study observes a previously neglected pharmacological phenomenon and investigates its mechanism of that the continuous low-dose administration of some antineoplastic agents in certain dose ranges can promote tumorigenesis and tumor progression in vitro and in vivo, through stimulation of tumor cell functions directly as well as enhancement of tumor angiogenesis by BMDCs recruitment indirectly. The results alert to a potential risk in current empirically based continuous low-dose chemotherapy regimens such as metronomic chemotherapy. There are indications that certain antineoplastic agents at low dosages may exhibit abnormal pharmacological actions, such as promoting tumor growth. However, the phenomenon still needs to be further confirmed, and its underlying mechanisms have not yet been fully elucidated. Gemcitabine (GEM) and cisplatin (CDDP) were employed as representative antineoplastic agents to observe effects of continuous low-dose chemotherapy with GEM or GEM combined with CDDP (GEM+CDDP) on tumor formation and growthin xenograft tumor models in vivo. Tumor and endothelial cell functions, apoptosis, cell cycle analysis, as well as bone marrow derived cells (BMDCs) mobilization, were evaluated with transwell, MTT or flow cytometry analysis in vitro, respectively. Histological methods were employed to assess angiogenesis in tumor tissues. The results showed that tumor formation and growth were both significantly promoted by GEM or GEM+CDDP at as low as half of the metronomic dosages, which were accompanied by enhancements of angiogenesis in tumor tissues and the release of proangiogenic BMDCs in the circulating blood. Additionally, GEM or GEM+CDDP at low concentrations dramatically facilitated the proliferation, migration, and invasion of tumor cells in vitro. Cell-cycle arrest, activation of associated apoptotic proteins, and inhibition of apoptosis were also observed in tumor cells. These findings indicate that, the continuous low-dose administration of GEM and GEM+CDDP can promote tumorigenesis and tumor progression in vivo by inhibiting apoptosis, mobilizing BMDCs, and promoting angiogenesis in certain dose ranges. These findings urge further investigations to avoid the potential risks in current empiric continuous low-dose chemotherapy regimens with antineoplastic agents.


Clinical Importance of Plasma Drug Concentration of Oral Molecular Targeted Drugs for Renal Cell Carcinoma.
Therapeutic drug monitoring (TDM) is widely used in clinical practice to maximize drug efficacy and minimize toxicities. Currently, it is also practiced in the use of oral molecular targeted drugs. The objective of this study was to assess the clinical importance of measuring the systemic concentration of oral molecular targeted drugs used to treat renal cell carcinoma (RCC). The systemic concentrations of the oral molecular targeted drugs sorafenib, sunitinib, axitinib, pazopanib, and everolimus used for RCC were useful for therapeutic interventions, and clinical outcomes were evaluated retrospectively. The interventional use of systemic drug concentration was confirmed in 26 of 87, and their categories are presented. The systemic concentration of sunitinib was useful in dose reduction and/or discontinuation (n = 10), dose escalation (n = 3), and adherence monitoring (n = 2). Nine of the 10 patients whose dose was reduced showed reduced adverse event. Two patients who were intervened in adherence monitor showed improved adherence. For axitinib, dose reduction and/or discontinuation (n = 1) and dose escalation (n = 6) were confirmed. For pazopanib, dose reduction and/or discontinuation (n = 1) and drug interaction detection (n = 1) were confirmed, both of them were confirmed to have reduced adverse events. For everolimus, dose reduction and/or discontinuation (n = 1) and drug interaction detection (n = 1) were confirmed, a patient with reduced dose recovered from adverse events. Interventions for sorafenib were not identified. This study demonstrated that systemic concentrations of oral molecular targeted drugs for RCC were considered to be clinically useful for dose adjustment, monitoring of treatment adherence, and the detection of drug interactions. Moreover, this information could be successfully used to guide individualized therapy to maximize the antitumor effects of these drugs.


Donor-type activated natural killer cells promote marrow engraftment and B cell development during allogeneic bone marrow transplantation.
Purified NK cells were obtained from mice with severe combined immune deficiency and were activated with human IL-2 (hrIL-2) in vitro to determine if, once activated, these cells could be transferred with compatible bone marrow cells (BMC) and promote marrow engraftment in irradiated allogeneic recipients. After culture with hrIL-2, these cells maintained a phenotypic and lytic spectrum consistent with a pure population of activated NK cells. These activated NK cells were then adoptively transferred with the donor BMC and rhIL-2 into lethally irradiated allogeneic hosts. The addition of NK cells with the BMC allowed for more rapid hematopoietic engraftment as determined through short term studies, and greater donor-derived chimerism with accelerated reconstitution of the B cell population as determined with long term analysis. No evidence of graft-vs-host disease was detected in the recipients receiving the activated NK cells with allogeneic T cell replete BMC and hrIL-2. The mechanism by which the transferred NK cells improved BMC engraftment was at least partly through the abrogation of the host effector cell's ability to mediate resistance to the marrow graft. Thus, the administration of donor-type activated NK cells with BMC and hrIL-2 may significantly augment hematopoietic engraftment and immune reconstitution in the clinical setting of allogeneic BMT without giving rise to graft-vs-host disease.


Sonic Landau Levels and Synthetic Gauge Fields in Mechanical Metamaterials.
Mechanical strain can lead to a synthetic gauge field that controls the dynamics of electrons in graphene sheets as well as light in photonic crystals. Here, we show how to engineer an analogous synthetic gauge field for lattice vibrations. Our approach relies on one of two strategies: shearing a honeycomb lattice of masses and springs or patterning its local material stiffness. As a result, vibrational spectra with discrete Landau levels are generated. Upon tuning the strength of the gauge field, we can control the density of states and transverse spatial confinement of sound in the metamaterial. We also show how this gauge field can be used to design waveguides in which sound propagates with robustness against disorder as a consequence of the change in topological polarization that occurs along a domain wall. By introducing dissipation, we can selectively enhance the domain-wall-bound topological sound mode, a feature that may potentially be exploited for the design of sound amplification by stimulated emission of radiation (SASER, the mechanical analogs of lasers).


Development of a Risk Scoring System for Predicting Anastomotic Leakage Following Laparoscopic Rectal Cancer Surgery.
To develop a risk scoring system that can predict the incidence of anastomotic leakage after laparoscopic rectal cancer surgery. The clinical data of 387 patients with rectal cancer who underwent laparoscopic low anterior resection were retrospectively collected. Univariable and multivariable logistic regression analyses were used to evaluate independent risk factors for postoperative anastomotic leakage. A simplified points system was then developed based on the corresponding regression coefficient β of each risk factor. Receiver operating characteristic (ROC) analysis was used to evaluate the performance and the optimal cut-off value in predicting anastomotic leakage. The performance of the points system was then externally validated in an independent cohort of 192 patients based in another institution. Anastomotic leakage occurred in 36 of 387 patients with rectal cancer (9.30%). Logistic multivariable regression analysis showed that males, maximum tumor diameter (≥5cm), operation time (≥180min), preoperative chemoradiation, intraoperative blood transfusion and the anastomosis level from the anal verge (≤5cm) were independent risk factors for the incidence of anastomotic leakage. According to the scoring standard, the risk points of each patient were calculated. ROC analysis based on the risk points showed that the area under the curve (AUC) was 0.795 (95% CI:0.752-0.834) and the optimal cut-off value was 6, yielding a sensitivity of 88.89% and a specificity of 62.96%. Using this risk points system, the AUC of another cohort of 192 patients from another institution who underwent laparoscopic low anterior resection for rectal cancer was 0.853 (95% CI:0.794-0.900, p<0.001) and patients with risk points ≥6 had a 21.05% chance of developing anastomotic leakage. This risk points system for predicting anastomotic leakage following laparoscopic rectal cancer surgery may be useful for surgeons in their decisions to perform intraoperative diversion stoma, which can reduce the incidence of postoperative anastomotic leakage.


The Asian species of strychnos. Part IV. The alkaloids.
Two hundred thirty-four samples from thirty-six Asian species of Strychnos have been screened for the presence of tertiary alkaloids; the data are set out in table 1. Thin-layer and gas-liquid chromatographic as well as mass-spectrometric studies have enabled alkaloids to be identified in many of the extracts prepared; the data are listed in table 2. The literature on the alkaloids and pharmacology of Asian Strychnos species is reviewed (altogether 99 plant names are considered) and compared with the findings obtained in the present work. The alkaloid economy of the plants is also discussed.


Defensive Functioning Moderates the Effects of Nondirective Meditation.
We have recently found that nondirective meditation facilitates stress reduction. This supplementary study investigated whether defensive functioning would moderate these beneficial effects. We explored the occurrence of defense mechanisms and the impact of defensive functioning on the outcome of companies' stress management programs regarding worries nervousness, mental distress, sleep problems, and muscle pain. The sample was a population of active, working professionals recruited from Norwegian companies (n = 105). The intervention group obtained significant benefits on all outcome measures, but there were no effects in the control group. We analyzed defensive functioning with the self-report questionnaire, Life Style Index, at four time points. The healthy adults who participated had a low level of defense scores at the outset. There was a significant reduction in the level of defenses in both groups over the study period, 6 months. Defensive functioning significantly moderated the change of the outcome measures from baseline to follow-up in the intervention group, but not in the control group.


Spectrophotometric and liquid-chromatographic studies of thymolphthalein monophosphate. Specifications for high-quality substrate for the measurement of prostatic acid phosphatase activity.
Fourteen lots of thymolphthalein monophosphate (TMP), disodium salt, obtained from 10 commercial suppliers were compared spectrophotometrically at 445 and 595 nm, liquid-chromatographically with monitoring at 254 nm, and enzymically by measurements of activity of prostatic acid phosphatase in human serum. Eight lots were classified as "unacceptable," six as "acceptable." Spectrophotometric testing revealed four lots with excessive thymolphthalein and three lots with grossly deficient amounts of TMP. In general, the chromatographic results paralleled those obtained by spectrophotometry, and both results correlated well with enzymic activity. Changing water content in this hygroscopic salt was a major problem, which resulted in great uncertainty as to the formula weight and therefore as to the moles of TMP actually taken. From these studies, specifications for high-quality TMP were determined. The critical importance of simultaneous enzymic activity measurements in comparisons with other "acceptable" lots in defining an adequate TMP substrate is stressed. Use of these specifications for selecting TMP for acid phosphatase activity measurements should improve intra- and inter-laboratory analytical performance.


Multilocus sequence typing (MLST) and whole-genome MLST of Campylobacter jejuni isolates from human infections in three districts during a seasonal peak in Finland.
A total of 95 human Campylobacter jejuni isolates acquired from domestic infections and collected from three districts in Finland during the seasonal peak (June to September) in 2012 were analyzed by PCR-based multilocus sequence typing (MLST) and by whole-genome sequencing (WGS). Four predominant sequence types (STs) were detected among the isolates: ST-45 (21%) and ST-230 (14%, ST-45 clonal complex [CC]), ST-267 (21%, ST-283 CC), and ST-677 (19%, ST-677 CC). In districts 1 and 3, most of the infections occurred from early July to the middle of August, with a peak at weeks 29 to 31, but in district 2, the infections were dispersed more evenly throughout 3 months (June to August). WGS data were used for further whole-genome MLST (wgMLST) analyses of the isolates representing the four common STs. Shared loci of the isolates within each ST were analyzed as distance matrices of allelic profiles by the neighbor-net algorithm. The highest allelic variations (>400 different alleles) were detected between different clusters of ST-45 isolates (1,121 shared loci), while ST-230 (1,264 shared loci), ST-677 (1,169 shared loci), and ST-267 isolates (1,217 shared loci) were less diverse with the clusters differing by <40 alleles. Closely related isolates showing no allelic variation (subclusters) were detected among all four major STs. In some cases, they originated from different districts, suggesting that isolates can be epidemiologically connected and may have the same infection source despite being originally identified as sporadic infections.


Translational regulation of the gradual increase in histone H1 kinase activity in maturing mouse oocytes.
In maturing mouse oocytes, p34cdc2-associated histone H1 kinase activity gradually increases until it reaches its maximum at metaphase I (Choi et al., 1991: Development 113:789-795). In this study, treatment of oocytes with cycloheximide resulted in a failure to increase the level of histone H1 activity above that detected at approximately the time of germinal vesicle breakdown (GVB), which is approximately 20-30% of the level normally achieved at metaphase I. Cyclin B was detected in GV-stage oocytes, but there was a 2-2.5-fold increase in the amount of cyclin B in maturing oocytes from GV-stage to metaphase I and a burst of cyclin B synthesis during the first 3 hr of maturation. Okadaic acid-treatment of mouse oocytes did not accelerate activation of histone H1 kinase but rather arrested its activity at the same level observed in cycloheximide-treated oocytes. Thus the components of the p34cdc2 kinase activating system in mouse oocytes are apparently not present in GV-stage oocytes in an amount or configuration that would allow maximum kinase activation when meiosis is reinitiated by okadaic acid. Importantly, okadaic acid-treatment dramatically inhibited protein synthesis. Therefore, the inhibition of protein synthesis by okadaic acid probably abrogates the possibility of de novo synthesis of the regulators of p34cdc2 kinase required to drive its activity to the maximum level normally achieved by metaphase I. It is concluded that there is a critical point in driving the continued activation of histone H1 kinase that occurs at approximately the time of GVB. Progression beyond this point requires de novo protein synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)


Detection of Zhang-Rice singlets using spin-polarized photoemission.
From a spin-resolved photoemission study on the Bi(2)Sr(2)CaCu(2)O(8+delta) superconductor, we show experimentally that the first ionization state is of nearly pure singlet character. This is true both above and below the superconducting transition and in the presence of doping and band formation. This provides direct support for the existence and stability of Zhang-Rice singlets in high-temperature superconductors, justifying the ansatz of single-band models. Moreover, we establish this technique as an important probe for a wide range of cuprates and strongly correlated materials.


[The effect of morphine on the input neuronal system of the spinal cord, participating in the formation of nociceptive pressor reflexes].
Effect of morphine applied to the spinal cord segments L4-S2 or C6-tI on pressor reflexes evoked by supramaximal stimulation of radial and tibial nerve with low frequency (I-2 Hz) was studied in anesthetized cats. Only pressor reflexes elicited by excitation of the tibial nerve afferents were suppressed by morphine applied to the L4-S2 segments. This effect was characterized by diminution of amplitude and steepness of the reflexes and by augmentation of their latency. Both the degree and the rate of the reflex suppression were found to enhance with increasing of morphine concentration from 0.02 to 0.5%. When applied to C6-tI segments, morphine did not suppress the pressor reflexes to the tibial nerve stimulation while reflexes to the radial nerve signals were decreased considerably. In addition to this local action of morphine, the effects resulting from it's distant action, namely, some reduction in systemic arterial pressure and an increase of pressor reflexes evoked by afferent signals entering into the spinal cord segments remote from the application region, were found to occur. All these effects were reversed by naloxone (0.2 mg/kg i. v.). It is concluded that along with attenuation of different withdrawal components of the defence reaction, action of morphine on the opiate receptors of some neurons situated near the entrance of afferent signals into the spinal cord results in suppression of the circulatory components of this reaction.


Childhood outcomes of assisted reproductive technology.
There is a large population of children conceived via assisted reproductive technology (ART), which continues to increase worldwide, without a clear understanding of associated long-term outcomes. ART children are more likely to be the result of multiple pregnancies, and thus to be born prematurely or low birthweight. There is growing evidence that ART children are phenotypically and biochemically different from naturally conceived children, but the mechanism(s) leading to these changes have not been elucidated. There is a possible increased risk of rare imprinted gene disorders in these children. However, it remains unclear whether more subtle changes in DNA methylation occur commonly, leading to differences in gene expression and phenotype in ART children. Although an increased risk of cancer among ART children has been reported, the role of ART in the development of cancer has not been demonstrated. Further research and ongoing surveillance of ART children is essential to better understand the possible effects of ART on the long-term health of this population.