Mutagenesis of the phosphatase sequence motif in diacylglycerol pyrophosphate phosphatase from Saccharomyces cerevisiae.
Diacylglycerol pyrophosphate (DGPP) phosphatase, encoded by the DPP1 gene, is a membrane-associated enzyme in the yeast Saccharomyces cerevisiae. The enzyme removes the beta phosphate from DGPP to form phosphatidate. The substrate and product of the DGPP phosphatase reaction play roles in lipid signaling and in cell metabolism. The deduced primary structure of the DGPP phosphatase protein contains a three-domain phosphatase sequence motif. In this work, we examined the hypothesis that the phosphatase sequence motif in the enzyme is involved in the DGPP phosphatase reaction. The amino acid residues Arg(125), His(169), and His(223) in domains 1, 2, and 3, respectively, of the phosphatase sequence motif were changed to alanine residues by site-directed mutagenesis. The mutant DPP1(R125A), DPP1(H169A), and DPP1(H223A) alleles were cloned into a yeast shuttle vector and then expressed in a dpp1Delta lpp1Delta double mutant that lacks DGPP phosphatase activity. Northern blot and immunoblot analyses showed that the mutations in the phosphatase sequence motif did not affect the expression of the enzyme. The DGPP phosphatase activities of the R125A, the H169A, and the H223A mutant enzymes were 0.05, 9, and 0.03%, respectively, of the DGPP phosphatase activity of the wild-type enzyme. Enzymes with mutations in more than one domain of the phosphatase sequence motif had no measurable DGPP phosphatase activity. The R125A and H233A mutant DGPP phosphatase enzymes had reduced V(max) and elevated K(m) values for DGPP when compared with the wild-type enzyme. The H169A mutant enzyme had reduced V(max) and K(m) values when compared with the control. The specificity constants (V(max)/K(m)()) for DGPP of the R125A mutant and H233A mutant enzymes were 4610-fold and 15 367-fold lower, respectively, when compared to the wild-type enzyme. The studies reported here indicated that the phosphatase sequence motif played an important role in the reaction catalyzed by the S. cerevisiae DGPP phosphatase.


Randomized Phase II Trial of Nivolumab Versus Nivolumab and Ipilimumab for Recurrent or Persistent Ovarian Cancer: An NRG Oncology Study.
Single-agent PD-1 blockade exhibits limited efficacy in epithelial ovarian cancer (EOC). We evaluated ipilimumab plus nivolumab compared with nivolumab alone in women with persistent or recurrent EOC. Eligibility criteria included measurable disease, 1-3 prior regimens, and platinum-free interval (PFI) < 12 months. Participants were randomly allocated to intravenous nivolumab (every 2 weeks) or induction with nivolumab plus ipilimumab for 4 doses (every 3 weeks), followed by every-2-week maintenance nivolumab for a maximum of 42 doses. The primary null hypothesis was equal probability of objective response within 6 months of random allocation in each arm. One hundred patients were allocated to receive either nivolumab (n = 49), or nivolumab plus ipilimumab (n = 51), with PFI of < 6 months in 62%. Six (12.2%) responses occurred within 6 months in the nivolumab group and 16 (31.4%) in the nivolumab plus ipilimumab group (odds ratio, 3.28; 85% CI, 1.54 to infinity; P = .034). The median progression-free survival (PFS) was 2 and 3.9 months in the nivolumab and nivolumab plus ipilimumab groups, respectively, with a PFI-stratified hazard ratio of 0.53 (95% CI, 0.34 to 0.82); the respective hazard ratio for death was 0.79 (95% CI, 0.44 to 1.42). Grade ≥ 3 related adverse events occurred in 33% of patients in the nivolumab group and 49% in the combination group, with no treatment-related deaths. PD-L1 expression was not significantly associated with response in either treatment group. Compared with nivolumab alone, the combination of nivolumab and ipilimumab in EOC resulted in superior response rate and longer, albeit limited, PFS, with toxicity of the combination regimen comparable to prior reports. Additional combination studies to enhance durability of the dual regimen are warranted.


An exploratory study on equity in funding allocation for essential medicines and health supplies in Uganda's public sector.
To ascertain equity in financing for essential medicines and health supplies (EMHS) in Uganda, this paper explores the relationships among government funding allocations for EMHS, patient load, and medicines availability across facilities at different levels of care. We collected data on EMHS allocations and availability of selected vital medicines from 43 purposively sampled hospitals and the highest level health centers (HC IV), 44 randomly selected lower-level health facilities (HC II, III), and from over 400 facility health information system records and National Medical Stores records. The data were analyzed to determine allocations per patient within and across levels of care and the effects of allocations on product availability. EMHS funding allocations per patient varied widely within facilities at the same level, and allocations per patient between levels overlapped considerably. For example, HC IV allocations per patient ranged from US$0.25 to US$2.14 (1:9 ratio of lowest to highest allocation), and over 75 % of HC IV facilities had the same or lower average allocation per patient than HC III facilities. Overall, 43 % of all the facilities had optimal stock levels, 27 % were understocked, and 30 % were overstocked. Using simulations, we reduced the ratio between the highest and lowest allocations per patient within a level of care to less than two and eliminated the overlap in allocation per patient between levels. Inequity in EMHS allocation is demonstrated by the wide range of funding allocations per patient and the corresponding disparities in medicines availability. We show that using patient load to calculate EMHS allocations has the potential to improve equity significantly. However, more research in this area is urgently needed. The article does not report any results of human participants. It is implemented in collaboration with the Uganda's Ministry of Health, Pharmacy Division.


A national study of fluoride mouthrinse adoption: implications for school health personnel.
The ongoing adoption of school-based fluoride mouthrinse programs in the U.S. has provided the opportunity to study the issues surrounding the adoption and implementation of a health technology by public schools. Findings from a National Study on the Diffusion of Preventive Health Measures to Schools have been selected for analysis with regard to the role of school health personnel in the adoption and delivery of the service. A more visible role for school health personnel as on-site health experts is discussed, as well as the potential ways for increasing their involvement through facilitating initial program adoption, relaying accurate and up-to-date implementation information to decision-makers, developing a constituency for long-term program support and ensuring program accuracy through monitoring and continuing education.


Telemedicine--the diagnostic tool of the future.
The most immediate concern in the development of telemedicine programs, technology, and equipment is the continuation of federal funding. As Congress continues its efforts to balance this year's federal budget, funds for research generally and particularly for the pilot programs discussed above could be cut or eliminated. Federal funding is surprisingly important to the rapid maturation of telemedicine. Without these pilot projects, HCFA believes that it would be unable to assess the effect of Medicare reimbursement for telemedicine consultations. Without such assessment, HCFA may be unwilling to relax the current restrictions on reimbursement for telemedicine outside of pilot programs. HMOs and third-party payors would then be less likely to support telemedicine programs and reimburse providers who participate in them. Similarly, without the promise of federal or private reimbursement, the telecommunications industry will find it harder to justify continuing research and development in new technologies. Nonetheless, even a complete elimination of all federal research monies is likely only to slow the growth of telemedicine, not stop it.


Sphingosine 1-Phosphate: A Novel Target for Lung Disorders.
Sphingosine 1-phosphate (S1P) is involved in a wide range of cellular processes, which include proliferation, apoptosis, lymphocyte egress, endothelial barrier function, angiogenesis, and inflammation. S1P is produced by two isoenzymes, namely, sphingosine kinase 1 and 2 (SphK1 and 2) and once produced, S1P can act both in an autocrine and paracrine manner. S1P can be dephosphorylated back to sphingosine by two phosphatases (SGPP 1 and 2) or can be irreversibly cleaved by S1P lyase. S1P has a diverse range of functions, which is mediated in a receptor dependent, through G-protein coupled receptors (S1PR1-5) or receptor independent manner, through intracellular targets such as HDACs and TRAF2. The involvement of S1P signaling has been confirmed in various disease conditions including lung diseases. The SphK inhibitors and S1PR modulators are currently under clinical trials for different pathophysiological conditions. There is a significant effort in targeting various components of S1P signaling for several diseases. This review focuses on the ways in which S1P signaling can be therapeutically targeted in lung disorders.


Gamma-interferon is the factor in lymphokine that activates human macrophages to inhibit intracellular Chlamydia psittaci replication.
We have demonstrated previously that mitogen-induced lymphokines activate human monocyte-derived macrophages to inhibit the intracellular replication of Chlamydia psittaci. To identify the factor(s) in crude lymphokines responsible for this antimicrobial effect, we tested human Con A-induced lymphokines for interferon activity. We also attempted to neutralize the lymphokines with a monoclonal antibody directed against human gamma-interferon and examined the ability of partially purified human gamma-interferon to induce macrophage antichlamydial activity. The lymphokine-induced antichlamydial effect was measured by the inhibition of chlamydial inclusion formation in Giemsa-stained macrophage cultures. Our lymphokines were found to be rich in gamma-interferon; treatment of cells for 48 hr before infection with lymphokines containing 300 U/ml of interferon resulted in an 89% inhibition of chlamydial growth. This lymphokine effect was completely abolished by monoclonal antibody against human gamma-interferon, but not by antisera against human alpha- or beta-interferons. In addition, partially purified human gamma-interferon alone induced macrophages to restrict chlamydial growth by 95%. We conclude that it is the gamma-interferon present in human Con A-induced lymphokines that activates monocyte-derived macrophages to inhibit chlamydial replication.


The potential of Fas ligand (apoptosis-inducing molecule) as an unconventional therapeutic target in type 1 diabetes.
The development of type 1 diabetes (T1D) is driven by autoreactive T cells that attack and destroy the insulin-producing β-cells in pancreatic islets, forcing patients to take multiple daily insulin injections. Insulin therapy, however, is not a cure and diabetic patients often develop serious long-term microvascular and cardiovascular complications. Therefore, intensive efforts are being directed toward developing safe immunotherapy for the disease that does not impair host defense and preserves β-cells, leading to better glycemic control than exogenous insulin therapy. Engineering therapies that differentially cripple or tolerate autoreactive diabetogenic T cells while sparing protective T cells necessary for maintaining a competent immune system has proven challenging. Instead, recent efforts have focused on modulating or resetting the immune system through global but transient deletion of T cells or B cells using anti-CD3 or anti-CD20 mAb, respectively. However, phase III clinical trials have shown promising but modest efficacy so far with these approaches. Therefore, there is a need to identify novel biological targets that do not fit the classic properties of being involved in adaptive immune cell activation. In this prospective, we provide preclinical evidence that targeting Fas ligand (FasL) may provide a unique opportunity to prevent or cure T1D and perhaps other organ-specific autoimmune diseases without causing immune suppression. Unlike conventional targets that are involved in T and B lymphocyte activation (such as CD3 and CD20, respectively), FasL is an apoptosis-inducing surface molecule that triggers cell death by binding to Fas (also known as CD95 Apo-1). Therefore, targeting FasL is not expected to cause immune suppression, the Achilles Heel of conventional approaches. We will discuss the hypothesis that targeting FasL has unique benefits that are not offered by current immunomodulatory approaches.


Interplay between tight junctions & adherens junctions.
Cell-cell adhesions are critical for the development and maintenance of tissues. Present at sites of cell-cell contact are the adherens junctions and tight junctions. The adherens junctions mediate cell-cell adhesion via the actions of nectins and cadherins. The tight junctions regulate passage of ions and small molecules between cells and establish cell polarity. Historically, the adherens and tight junctions have been thought of as discrete complexes. However, it is now clear that a high level of interdependency exists between the two junctional complexes. The adherens junctions and tight junctions are physically linked, by the zonula occludens proteins, and linked via signaling molecules including several polarity complexes and actin cytoskeletal modifiers. This review will first describe the individual components of both the adherens and tight junctions and then discuss the coupling of the two complexes with an emphasis on the signaling links and physical interactions between the two junctional complexes.


Rehabilitation facilitates functional improvement following intravenous infusion of mesenchymal stem cells in the chronic phase of cerebral ischemia in rats.
The primary objective of this study was to investigate the potential facilitating effects of daily rehabilitation for chronic cerebral ischemia following the intravenous infusion of mesenchymal stem cells (MSC) in rats. The middle cerebral artery (MCA) was occluded by intraluminal occlusion using a microfilament (MCAO). Eight weeks after MCAO induction, the rats were used as a chronic cerebral ischemia model. Four experimental groups were studied: Vehicle group (medium only, no cells); Rehab group (vehicle + rehabilitation), MSC group (MSC only); and Combined group (MSC + rehabilitation). Rat MSCs were intravenously infused eight weeks after MCAO induction, and the rats received daily rehabilitation through treadmill exercise for 20 min. Behavioral testing, lesion volume assessment using magnetic resonance imaging (MRI), and histological analysis were performed during the observation period until 16 weeks after MCAO induction. All treated animals showed functional improvement compared with the Vehicle group; however, the therapeutic efficacy was greatest in the Combined group. The combination therapy is associated with enhanced neural plasticity shown with histological analysis and MRI diffusion tensor imaging. These findings provide behavioral evidence for enhanced recovery by combined therapy with rehabilitation and intravenous infusion of MSCs, and may form the basis for the development of clinical protocols in the future.


Effect of pinealectomy on serum melatonin, luteinizing hormone and prolactin: a case report.
A patient, aged 47 is described in whom changes in hormonal profiles were observed following surgical excision of the pineal. She presented with a clinical syndrome of increased intracranial pressure and postmenopausal symptoms. Computerized tomographic scans showed obstructive hydrocephalus from a cystic lesion in the pineal region. Surgical removal of the cystic mass revealed a benign ependymal cyst and normal pineal tissue. After the operation, she had no nocturnal increase in serum melatonin. The morning basal LH was further elevated and night-time PRL showed little fluctuation from the basal levels.


TRPM2 cation channels, oxidative stress and neurological diseases: where are we now?
The Na+ and Ca(2+)-permeable melastatin related transient receptor potential 2 (TRPM2) channels can be gated either by ADP-ribose (ADPR) in concert with Ca(2+) or by hydrogen peroxide (H(2)O(2)), an experimental model for oxidative stress, binding to the channel's enzymatic Nudix domain. Since the mechanisms that lead to TRPM2 gating in response to ADPR and H(2)O(2) are not understood in neuronal cells, I summarized previous findings and important recent advances in the understanding of Ca(2+) influx via TRPM2 channels in different neuronal cell types and disease processes. Considering that TRPM2 is activated by oxidative stress, mediated cell death and inflammation, and is highly expressed in brain, the channel has been investigated in the context of central nervous system. TRPM2 plays a role in H(2)O(2) and amyloid β-peptide induced striatal cell death. Genetic variants of the TRPM2 gene confer a risk of developing Western Pacific amyotropic lateral sclerosis and parkinsonism-dementia complex and bipolar disorders. TRPM2 also contributes to traumatic brain injury processes such as oxidative stress, inflammation and neuronal death. There are a limited number of TRPM2 channel blockers and they seem to be cell specific. For example, ADPR-induced Ca(2+) influx in rat hippocampal cells was not blocked by N-(p-amylcinnomoyl)anthralic acid (ACA), the IP(3) receptor inhibitor 2-aminoethoxydiphenyl borate or PLC inhibitor flufenamic acid (FFA). However, the Ca(2+) entry in rat primary striatal cells was blocked by ACA and FFA. In conclusion TRPM2 channels in neuronal cells can be gated by either ADPR or H(2)O(2). It seems to that the exact relationship between TRPM2 channels activation and neuronal cell death still remains to be determined.


The effect of El Niño and La Niña episodes on the existing niche and potential distribution of vector and host species of American Cutaneous Leishmaniasis.
Leishmaniasis is a zoonotic disease transmitted to humans by a protozoan parasite through sandfly vectors and multiple vertebrate hosts. The Pan American Health Organization reported a declining trend in cases, with Brazil, Colombia, Peru, Nicaragua, and Bolivia having the most cases in 2020. There are still knowledge gaps in transmission and the parasite-host relationship. Ecological niche modeling has been used to study host-vector relationships, disease dynamics, and the impact of climate change. Understanding these aspects can aid in early surveillance and vector control strategies. The potential distribution of five host species associated with the transmission of cutaneous leishmaniasis (CL) was modeled. Occurrence data were collected for each host species, and environmental variables were used to build the models. Climatic data from El Niño, La Niña, and Neutral episodes were used to compare the predicted distributions. Additionally, the potential distributions of four vector species were compared to identify overlaps with host species. Niche analysis was conducted to evaluate changes in vector niches across episodes and to identify host-vector pairs based on niche overlap in geographic and environmental spaces. After spatial thinning, 467 records were obtained, and 1,190 candidate models were evaluated for each species. Results showed the distribution of occurrences in the environmental space, highlighting a high risk of extrapolation beyond the calibration areas. Movement-Oriented Parity analysis revealed distinct distribution patterns under different climate conditions, with areas of environmental similarity identified. Bradypus variegatus exhibited a broad potential distribution, while Dasypus novemcinctus and Didelphis marsupialis had more restricted ranges. Sylvilagus braziliensis covered most of the Neotropics. Our study provides valuable insights into ecological niches and geographic ranges of these species, contributing to the understanding of cutaneous leishmaniasis transmission dynamics.


Wide-field endoscopic mucosal resection versus endoscopic submucosal dissection for laterally spreading colorectal lesions: a cost-effectiveness analysis.
To compare the cost-effectiveness of endoscopic submucosal dissection (ESD) and wide-field endoscopic mucosal resection (WF-EMR) for removing large sessile and laterally spreading colorectal lesions (LSLs) >20 mm. An incremental cost-effectiveness analysis using a decision tree model was performed over an 18-month time horizon. The following strategies were compared: WF-EMR, universal ESD (U-ESD) and selective ESD (S-ESD) for lesions highly suspicious for containing submucosal invasive cancer (SMIC), with WF-EMR used for the remainder. Data from a large Western cohort and the literature were used to inform the model. Effectiveness was defined as the number of surgeries avoided per 1000 cases. Incremental costs per surgery avoided are presented. Sensitivity and scenario analyses were performed. 1723 lesions among 1765 patients were analysed. The prevalence of SMIC and low-risk-SMIC was 8.2% and 3.1%, respectively. Endoscopic lesion assessment for SMIC had a sensitivity and specificity of 34.9% and 98.4%, respectively. S-ESD was the least expensive strategy and was also more effective than WF-EMR by preventing 19 additional surgeries per 1000 cases. 43 ESD procedures would be required in an S-ESD strategy. U-ESD would prevent another 13 surgeries compared with S-ESD, at an incremental cost per surgery avoided of US$210 112. U-ESD was only cost-effective among higher risk rectal lesions. S-ESD is the preferred treatment strategy. However, only 43 ESDs are required per 1000 LSLs. U-ESD cannot be justified beyond high-risk rectal lesions. WF-EMR remains an effective and safe treatment option for most LSLs. NCT02000141.


Relationship between neighborhood walkability and older adults' physical activity: results from the Belgian Environmental Physical Activity Study in Seniors (BEPAS Seniors).
Adequate knowledge on environmental correlates of physical activity (PA) in older adults is needed to develop effective health promotion initiatives. However, research in this age group is scarce and most existing studies were conducted in North America. The present study aimed to examine relationships between GIS-based neighborhood walkability and objective and self-reported PA in community-dwelling Belgian older adults. Furthermore, moderating effects of neighborhood income levels were investigated. The Belgian Environmental Physical Activity Study (BEPAS) for Seniors is a cross-sectional study in older adults (≥65 yrs) and was conducted between October 2010 and September 2012. Data from 438 older adults living in 20 neighborhoods across Ghent (Belgium) were analyzed. Stratification of selected neighborhoods was based upon objective walkability and neighborhood income. Participants wore an accelerometer during seven consecutive days to obtain objective levels of moderate-to-vigorous physical activity (MVPA). Self-reported levels of transportation walking/cycling and recreational walking/cycling were assessed using the International Physical Activity Questionnaire (long, last 7 days version) adapted for the elderly. Multi-level regression analyses were conducted. Findings showed a positive relationship between neighborhood walkability and weekly minutes of older adults' self-reported walking for transportation (B = 4.63 ± 1.05;p < 0.001) and a negative relationship between walkability and accelerometer-derived low-light PA (B = -1.38 ± 0.62;p = 0.025). Walkability was not related to any measure of recreational PA. A walkability x income interaction was found for accelerometer-derived MVPA (B = -1.826 ± 1.03;p = 0.075), showing only a positive association between walkability and MVPA in low-income neighborhood residents. This was the first European study to examine walkability-PA relationships in older adults. These Belgian findings suggest that a high neighborhood walkability relates to higher levels of older adults' transport-related walking. As transport-related walking is an accessible activity for older adults and easy to integrate in their daily routine, policy makers and health promoters are advised to provide sufficient destinations and pedestrian-friendly facilities in the close vicinity of older adults' residences, so short trips can be made by foot. Neighborhood income moderated the relationship between walkability and objectively-measured MVPA. Increasing total MVPA levels in older adults should be a key topic in development of promotion initiatives and special attention should be paid to low-income neighborhood residents.


Novel properties of hepatic canalicular reduced glutathione transport revealed by radiation inactivation.
Transport of GSH at the canalicular pole of hepatocytes occurs by a facilitative carrier and can account for approximately 50% of total hepatocyte GSH efflux. A low-affinity unit with sigmoidal kinetics accounts for 90% of canalicular transport at physiological GSH concentrations. A low-capacity transporter with high affinity for GSH has also been reported. It is not known whether the same or different proteins mediate low- and high-affinity GSH transport, although they do differ in inhibitor specificity. The bile of rats with a mutation in the canalicular multispecific organic anion transporter (cMOAT or MRP-2, a 170-kDa protein) is deficient in GSH, implying that cMOAT may transport GSH. However, transport of GSH in canalicular membrane vesicles (CMV) from these mutant rats remains intact. We examined the functional size of the two kinetic components of GSH transport by radiation inactivation of GSH uptake in rat hepatic CMV. High-affinity transport of GSH was inactivated as a single exponential function of radiation dose, yielding a functional size of approximately 70 kDa. In contrast, low-affinity canalicular GSH transport exhibited a complex biexponential response to irradiation, characterized by an initial increase followed by a decrease in GSH transport. Inactivation analysis yielded a approximately 76-kDa size for the low-affinity transporter. The complex inactivation indicated that the low-affinity transporter is associated with a larger protein of approximately 141 kDa, which masked approximately 80% of the potential transport activity in CMV. Additional studies, using inactivation of leukotriene C4 transport, yielded a functional size of approximately 302 kDa for cMOAT, indicating that it functions as a dimer.


Positron emission tomography and granulosa cell tumor recurrence: a report of 2 cases.
Two case reports of women with recurrent granulosa cell tumors identified initially by increasing levels of inhibin. As part of their investigation to assess the extent of the recurrence, an abdominopelvic computed tomography and a positron emission tomography scans were performed. Interestingly, the recurrent tumors were identified on the abdominopelvic computed tomography but not on the positron emission tomography scan. These recurrences were confirmed at surgery, and the histopathologic findings were identical to the original lesion.


Neprilysin inhibitors as a new approach in the treatment of right heart failure in the course of chronic obstructive pulmonary disease.
The aim of the study was to find out scientific evidence on the possible use of the combined angiotensin II receptor antagonist and neprilysin inhibitors (ARNI) in patients with right heart failure (RHF) in the course of chronic obstructive pulmonary disease (COPD). It has been proven that a lack of neprilysin or its reduced expression in hypoxia leads to exacerbation of pulmonary arterial remodelling (PAR) or pulmonary hypertension (PH) in the mechanism related to the platelet-derived growth factor (PDGF) resulting in the proliferation and migration of pulmonary artery smooth muscle cells and endothelial-to-mesenchymal transition. Such action in the course of COPD can lead to RHF, which would signify noxious effect of this group of drugs. However, the inhibition of neprilysin also inhibits natriuretic peptide metabolism. The representative of this group - brain natriuretic peptide (BNP) - acts as a vasodilator and also exerts an anti-proliferative activity through the cGMP-dependent protein kinase G pathway. Additionally, it causes bronchodilation by inducing the release of acetylcholine from bronchial epithelial cells. This suggests that natriuretic peptides may appear to be a potential treatment agent in patients with cardiac complications and COPD. Their effects associated with the immunosuppression capacity by reducing the release of inflammatory mediators - IL-6, IL-1β, and TNF-α can bring benefits to patients with acute lung injury caused by pulmonary inflammation during COPD exacerbations. Considering the potentially positive effect of natriuretic peptides in this group of patients, further research is required in this area, which can provide strong scientific data demonstrating the need for introducing ARNI drugs to the treatment of patients with COPD.


JMJ14, a JmjC domain protein, is required for RNA silencing and cell-to-cell movement of an RNA silencing signal in Arabidopsis.
JMJ14 is a histone H3 Lys4 (H3K4) trimethyl demethylase that affects mobile RNA silencing in an Arabidopsis transgene system. It also influences CHH DNA methylation, abundance of endogenous transposon transcripts, and flowering time. JMJ14 acts at a point in RNA silencing pathways that is downstream from RNA-dependent RNA polymerase 2 (RDR2) and Argonaute 4 (AGO4). Our results illustrate a link between RNA silencing and demethylation of histone H3 trimethylysine. We propose that JMJ14 acts downstream from the Argonaute effector complex to demethylate histone H3K4 at the target of RNA silencing.


Synchronous and metachronous carcinoma of the breast.
To determine the incidence of synchronous bilateral breast cancer, 100 women receiving their initial treatment for primary breast cancer prospectively underwent biopsy of the opposite breast. No patient had a clinically detectable abnormality in the contralateral breast, but 2% had unsuspected invasive ductal carcinomas. An additional 5% had premalignant lesions. To determine the incidence of the development of metachronous breast cancer, 153 women who underwent surgical treatment of primary breast cancer between 1955 and 1979 were followed from 2 months to 26 years, with an average of 3.6 years. During that time, 3.9% developed metachronous carcinomas of the contralateral breast.


CareTrack Kids-part 2. Assessing the appropriateness of the healthcare delivered to Australian children: study protocol for a retrospective medical record review.
Australian and international clinical practice guidelines are available for common paediatric conditions. Yet there is evidence that there are substantial variations between the guidelines, recommendations (appropriate care) and the care delivered. This paper describes a study protocol to determine the appropriateness of the healthcare delivered to Australian children for 16 common paediatric conditions in acute and primary healthcare settings. A random sample of 6000-8000 medical records representing a cross-section of the Australian paediatric population will be reviewed for appropriateness of care against a set of indicators within three Australian states (New South Wales, Queensland and South Australia) using multistage, stratified sampling. Medical records of children aged <16 years who presented with at least one of the study conditions during 2012 and 2013 will be reviewed. Human Research Ethics Committee approvals have been received from the Sydney Children's Hospital Network, Children's Health Queensland Hospital and Health Service and Women's and Children's Hospital Network (South Australia). An application is under review for the Royal Australian College of General Practitioners. The authors will submit the results of the study to relevant journals and offer oral presentations to researchers, clinicians and policymakers at national and international conferences.


Effects of PCB exposure on biochemical and hematological findings in capacitor workers.
Certain former operations in capacitor manufacturing resulted in extensive direct contact of the workers with electrical grade polychlorinated biphenyls (PCBs). A study group of 194 such individuals, all exposed to Aroclor 1016 and many previously exposed to Aroclors 1242 and/or 1254, was examined before (1976) and after (1979) discontinuance of PCB use in the operations (1977). At the two examinations, the approximate geometric mean serum levels (in ppb) and 5 to 95% ranges were for lower PCBs (LPCB), 363 (57-2270) and 68 (12-392); and for higher PCBs (HPCB), 30 (6-142) and 19 (4-108), respectively. The statistical associations among 42 measured clinical chemical and hematological parameters, five different measures of PCB exposure, and seven confounding variables observed in the two examinations were determined by three regression procedures. Similar regressions were performed with DDE, which was present at background levels. The principal statistical findings were a depression in serum bilirubin and elevations in serum GGTP and lymphocyte levels at the time of the first examination, and only an elevation in monocytes at the second. Appraisal of the results suggested an induction of microsomal enzymes which appeared to be subsiding after the cessation of direct exposure to PCBs. The statistical association between serum levels of PCBs and lipids reported by others was confirmed, but shown to be explained by the partitioning behavior of PCB in the body, rather than to changes in liver function. No evidence for health impairment related to PCBs was found, despite the high serum levels of PCBs in the study population.


Interpersonal Dyadic Influences of Pain Catastrophizing Between Caregivers and Children With Chronic Pain.
Pain catastrophizing is an important predictor of pain-related outcomes. Caregiver and child levels of catastrophizing about child chronic pain are associated cross-sectionally, yet predictive associations testing interpersonal influences within caregiver-child dyads are lacking. The present study tested caregiver and child influences on partner catastrophizing about child pain over a period of 1 month following initiation of interdisciplinary pain treatment and examined whether the change in pain catastrophizing was associated with child pain interference. A total of 113 caregiver-child dyads (Mage=14.41) completed measures at the time of initiating care at a pediatric tertiary outpatient pain management clinic (baseline) and ∼1 month later. Caregivers and children independently reported on catastrophizing about child pain and child pain interference at baseline and 1-month follow-up. Caregiver and child pain catastrophizing decreased over 1 month following initial interdisciplinary pain evaluation, with average scores remaining in the moderate to high range. Change in caregiver, but not child, catastrophizing about child pain was predicted by partner baseline pain catastrophizing. Decreases in catastrophizing about child pain were associated with within-person improvement in ratings of child pain interference. In the short period following initial pain evaluation, caregivers and children evidenced reductions in pain catastrophizing, which were associated with increased child function. Findings highlight the important role of child cognitive-affective responses to pain in influencing caregiver catastrophizing about child pain. Understanding the individual contributions children and caregivers make to interpersonal pain processes will inform future family-level clinical interventions.


Lesser sac herniation through a defect in the transverse mesocolon: CT findings.
We herein report a case of lesser sac herniation through a defect in the transverse mesocolon. In addition to CT signs suggestive of bowel obstructions, there are some characteristic CT findings in this extremely rare type of internal herniation, which include clustering of distended jejunal segments directly compressing the pancreas, displacement of the stomach anterolaterally, displacement of the transverse colon anteriorly and a normal appearance around the foramen of Winslow.


V1 as an egocentric cognitive map.
We typically distinguish between V1 as an egocentric perceptual map and the hippocampus as an allocentric cognitive map. In this article, we argue that V1 also functions as a post-perceptual egocentric cognitive map. We argue that three well-documented functions of V1, namely (i) the estimation of distance, (ii) the estimation of size, and (iii) multisensory integration, are better understood as post-perceptual cognitive inferences. This argument has two important implications. First, we argue that V1 must function as the neural correlates of the visual perception/cognition distinction and suggest how this can be accommodated by V1's laminar structure. Second, we use this insight to propose a low-level account of visual consciousness in contrast to mid-level accounts (recurrent processing theory; integrated information theory) and higher-level accounts (higher-order thought; global workspace theory). Detection thresholds have been traditionally used to rule out such an approach, but we explain why it is a mistake to equate visibility (and therefore the presence/absence of visual experience) with detection thresholds.


[Shoulder arthroplasty in the private practice. Analysis of demographics and comorbid conditions in a sample of 120 patients].
To describe the demographics in a series of shoulder arthroplasty cases operated in a single hospital and by the same surgical team. 120 shoulder arthroplasties performed between January 2006 and November 2014 were included. The following variables were analyzed: age, gender, baseline diagnosis, type of prosthesis used, comorbid conditions, involved side, occupation, follow-up time, and surgical history. Females accounted for 66% and males 34%; mean age was 66 years and the most frequent diagnosis was arthropathy resulting from rotator cuff tear (30%). The reverse total shoulder prosthesis was used in 51% of cases, followed by the hemiprosthesis in 38%. Seventy per cent of patients were from the Federal District. Arterial hypertension occurred in 23% of cases; 36% of cases had undergone prior surgery; 40% of cases worked at home. Shoulder arthroplasty is a procedure frequently performed at this hospital. Our case series reports a significant number of cases in which the demographics may give us an overview of shoulder arthroplasty in Mexican patients and describe the characteristics of patients eligible for this procedure, as there are no other registries available.


Adoptive cell therapies for glioblastoma.
Glioblastoma (GBM) is the most common and most aggressive primary brain malignancy and, as it stands, is virtually incurable. With the current standard of care, maximum feasible surgical resection followed by radical radiotherapy and adjuvant temozolomide, survival rates are at a median of 14.6 months from diagnosis in molecularly unselected patients (1). Collectively, the current knowledge suggests that the continued tumor growth and survival is in part due to failure to mount an effective immune response. While this tolerance is subtended by the tumor being utterly "self," it is to a great extent due to local and systemic immune compromise mediated by the tumor. Different cell modalities including lymphokine-activated killer cells, natural killer cells, cytotoxic T lymphocytes, and transgenic chimeric antigen receptor or αβ T cell receptor grafted T cells are being explored to recover and or redirect the specificity of the cellular arm of the immune system toward the tumor complex. Promising phase I/II trials of such modalities have shown early indications of potential efficacy while maintaining a favorable toxicity profile. Efficacy will need to be formally tested in phase II/III clinical trials. Given the high morbidity and mortality of GBM, it is imperative to further investigate and possibly integrate such novel cell-based therapies into the current standards-of-care and herein we collectively assess and critique the state-of-the-knowledge pertaining to these efforts.


[Antigen-binding lymphocytes in acute myocardial infarct].
Sensitized lymphocytes capable of fixing the cardiac antigen and its components, myoglobin and actomyosin, are detected in acute myocardial infarction. Their amount increases particularly sharply in the first 3 or 4 weeks after the infarction, which is indicative of the high activity of the cell link of the autoimmune process.


In situ carbon turnover dynamics and the role of soil microorganisms therein: a climate warming study in an Alpine ecosystem.
Litter decomposition represents one of the largest fluxes in the global terrestrial carbon cycle. The aim of this study was to improve our understanding of the factors governing decomposition in alpine ecosystems and how their responses to changing environmental conditions change over time. Our study area stretches over an elevation gradient of 1000 m on the Hochschwab massif in the Northern Limestone Alps of Austria. We used high-to-low elevation soil translocation to simulate the combined effects of changing climatic conditions, shifting vegetation zones, and altered snow cover regimes. In original and translocated soils, we conducted in situ decomposition experiments with maize litter and studied carbon turnover dynamics as well as temporal response patterns of the pathways of carbon during microbial decomposition over a 2-year incubation period. A simulated mean annual soil warming (through down-slope translocation) of 1.5 and 2.7 °C, respectively, resulted in a significantly accelerated turnover of added maize carbon. Changes in substrate quantity and quality in the course of the decomposition appeared to have less influence on the microbial community composition and its substrate utilization than the prevailing environmental/site conditions, to which the microbial community adapted quickly upon change. In general, microbial community composition and function significantly affected substrate decomposition rates only in the later stage of decomposition when the differentiation in substrate use among the microbial groups became more evident. Our study demonstrated that rising temperatures in alpine ecosystems may accelerate decomposition of litter carbon and also lead to a rapid adaptation of the microbial communities to the new environmental conditions.


[Metabolic stress by guanosin in healthy persons and in patients suffering from gouty arthritis].
The oral administration of 2 g guanosine resulted in both normal and gouty patients in a significant increase of serum uric acid levels, remaining for 24 hours above the initial values. There was no difference in the response of both groups investigated, indicating that a "purine-tolerance test" has no significance in the diagnosis of arthritis urica. The 24-hour-rhythm of serum aicd was studied in 20 normal persons, showing no statistical significant differences. The results of this investigation demonstrate that blood-sampling for uric acid determination can be performed at any time of the day, provided a provided a purine-poor diet is given 24 hours before.


A randomized iterative approach to optimizing an online substance use intervention for collegiate athletes.
Rates of drug use among collegiate athletes are high, yet there are few evidence-based interventions for this population. myPlaybook, an online intervention for collegiate athletes, targets multiple predictors of drug use (i.e., norms, positive and negative expectancies about use, and harm prevention intentions). We aimed to optimize modules from myPlaybook. We evaluated modules through three sequential randomized factorial trials, using the Multiphase Optimization Strategy framework. We recruited and randomized 54 (Trial 1), 47 (Trial 2), and 42 (Trial 3) schools and invited all first-year and transfer collegiate athletes to participate. Athletes completed a baseline survey, their randomly assigned modules, and immediate posttest and 30-day follow-up surveys. Across trials, 3,244 (48.8% female), 2,837 (51.9% female), and 2,193 (51.4% female) athletes participated. In Trial 1, we evaluated and revised less effective modules (defined as d < 0.3-0.4 for targeted outcomes). In Trial 2, we re-evaluated and revised less effective modules. In Trial 3, we re-evaluated the revised modules. Trial 1: All effects were d < 0.15, so we revised modules to target proximal outcomes (i.e., the hypothesized mediating variables in our conceptual model), rather than specific drug use behaviors. Trial 2: Most effects were d < 0.3, so we revised all modules. Trial 3: The norms module improved descriptive and injunctive norms (all d >0.35). The expectancies module improved alcohol positive expectancies (d = 0.3). The other modules were not effective. After three trials, two myPlaybook modules substantially improved proximal outcomes, increasing the likelihood that the combined intervention will have a meaningful clinical impact on collegiate athletes' drug use.


Optimizing insulin therapy in patients with type 1 and type 2 diabetes mellitus: optimal dosing and timing in the outpatient setting.
Management of type 1 and type 2 diabetes is continually evolving, and among these evolving therapies is administration of insulin in its various forms. The insulin regimen needs to be tailored to each individual, not only to maximize compliance and glycemic control but also to minimize hypoglycemia and weight gain.


Development of a bedside tool to predict time to death after withdrawal of life-sustaining therapies in infants and children.
To generate a preliminary bedside predictor of rapid time-to-death after withdrawal of support in children to help identify potential candidates for organ donation after circulatory death. Retrospective chart review. Pediatric intensive care unit of an academic children's hospital. All deaths in the pediatric intensive care unit from May 1996 to April 2007. None. Among 1389 deaths, 634 patients underwent withdrawal of support and 518 with complete data regarding demographics, life-supportive therapies, and end-of-life circumstances were analyzed. Three hundred seventy-three (72%) patients died within 30 mins of withdrawal and 452 (87%) died within 60 mins. Using multiple logistic regression, significant predictors of death within 30 or 60 mins (typical cut-off times for organ donation) were identified and a predictor score was generated. Significant predictors included: age 1 month or younger; norepinephrine, epinephrine, or phenylephrine >0.2 µg/kg/min; extracorporeal membrane oxygenation; and positive end-expiratory pressure >10 cmH2O; and spontaneous ventilation. Possible scores for the 30-min predictor ranged from -17 to 67; a score ≤-9 predicted a 37% probability of death ≤ 30 mins, whereas a score ≥ 38 predicted an 85% probability of death within 30 mins. For the 60-min predictor, scores ranged from -21 to 38; score ≤-10 predicted a 59% probability of death within 60 mins and a score ≥ 16 predicted a 98% probability of death within 60 mins. This tool is a reasonable preliminary predictor for death within 30 or 60 mins after withdrawal of support in terminally ill or injured children and might assist in identifying potential pediatric candidates for donation after circulatory death, although prospective validation is required.


Understanding the role of inflammatory-related pathways in the pathophysiology and treatment of psychiatric disorders: evidence from human peripheral studies and CNS studies.
Many lines of evidence now support the hypothesis that inflammation-related pathways are involved in the pathophysiology of psychiatric disorders. Much of the data underpinning this hypothesis has come from the study of inflammation-related proteins in blood of individuals with mood disorders and schizophrenia. Significantly, recent data have emerged to suggest that changes in inflammation-related pathways are present in the CNS of subjects with psychiatric disorders. It is therefore timely to overview how such data, plus data on the role of inflammation-related proteins in CNS function, is contributing to understanding the pathophysiology of mood disorders and schizophrenia. In addition, it has been suggested that antidepressants, mood stabilizers and antipsychotic drugs act on inflammation-related pathways and therefore measuring levels of inflammation-related proteins in blood may be useful in monitoring treatment responsiveness. Despite these important neuropsychopharmacological discoveries, there is no clear understanding as to how inflammatory-related pathways can precipitate the onset of psychiatric symptoms. This review will focus on data suggesting that acute-reactive proteins and cytokines are affected by the pathophysiology of mood disorders and schizophrenia, that levels of blood inflammation-related proteins before and after treatment might be useful in the diagnosis of psychiatric disorders or measuring responsiveness to drug treatment. Finally, it will be postulated how changes in these proteins affect CNS function to cause psychiatric disorders.


Functional roles of the two domains of phosducin and phosducin-like protein.
Phosducin and phosducin-like protein regulate G protein signaling pathways by binding the betagamma subunit complex (Gbetagamma) and blocking Gbetagamma association with Galpha subunits, effector enzymes, or membranes. Both proteins are composed of two structurally independent domains, each constituting approximately half of the molecule. We investigated the functional roles of the two domains of phosducin and phosducin-like protein in binding retinal G(t)betagamma. Kinetic measurements using surface plasmon resonance showed that: 1) phosducin bound G(t)betagamma with a 2. 5-fold greater affinity than phosducin-like protein; 2) phosphorylation of phosducin decreased its affinity by 3-fold, principally as a result of a decrease in k(1); and 3) most of the free energy of binding comes from the N-terminal domain with a lesser contribution from the C-terminal domain. In assays measuring the association of G(t)betagamma with G(t)alpha and light-activated rhodopsin, both N-terminal domains inhibited binding while neither of the C-terminal domains had any effect. In assays measuring membrane binding of G(t)betagamma, both the N- and C-terminal domains inhibited membrane association, but much less effectively than the full-length proteins. This inhibition could only be described by models that included a change in G(t)betagamma to a conformation that did not bind the membrane. These models yielded a free energy change of +1.5 +/- 0.25 kcal/mol for the transition from the G(t)alpha-binding to the Pd-binding conformation of G(t)betagamma.


Is beauty in the eye of the beholder? Effects of weight and shape on attractiveness ratings of female line drawings by restrained and nonrestrained eaters.
This study tested the differences between restrained and nonrestrained eaters' attractiveness perceptions of female line drawings, of their own figures, and the ideal female figure. Female line drawings varied systematically in body weight and in waist and hip circumference. Forty-six female undergraduate students, 23 nonrestrainers and 23 restrainers, rated stimuli in attractiveness, identified the figure which best represented their own body type (PAF), and the ideal body figure (IF) according to the Restraint Scale [RS; Herman, C. P., & Polivy, J. (1980). Restrained eating. In: A. Stunkard (Ed.), Obesity (pp. 208-225). Philadelphia, PA: Saunders]. Restrainers did not generally differ from nonrestrainers in attractiveness ratings or in their choice of IF. However, differences between IF and PAF were larger in restrainers than in nonrestrainers because restrainers chose PAFs with larger hips than nonrestrainers did. This difference between the restraint groups was independent of between-group differences in hip size. This discrepancy between IF and PAF may contribute to the restrainers' motivation to diet.


Isolation and characterization of protein kinase C from rat brain synaptosome cytoskeleton.
Synaptosomes prepared from brain tissues are known to retain morphological and functional characteristics of the nerve ending. Little information is available, however, as to the biochemical events underlying synaptogenesis and transmitter release. Increasing body of evidence suggests that protein kinase C (PKC) plays crucially important roles through phosphorylation of membrane proteins such as GAP-43 (for 43-kDa growth-associated protein) and 87-kDa MARCKS (for myristoylated, alanine-rich C kinase substrate) in many aspects of the neuronal function. Among them, arrangement of membrane cytoskeletal protein is proposed to be one of the primary sites of PKC action. The present study is an attempt to isolate and characterize PKC associated with synaptosomal membrane cytoskeleton. Rat brain synaptosomal Triton X-100 insoluble elements (cytoskeleton) contains specific [3H]phorbol dibutyrate binding activity and 78-kDa protein which reacts with an antibody against beta II-PKC subspecies. Although 78-kDa protein could not be solublized by the treatment with various ionic and non-ionic detergents and/or high concentrations of salts such as NaCl and LiBr, the fragment of 78-kDa protein was produced and solublized from cytoskeleton by limited proteolysis with calpain II, which cleaves PKC at one or two specific sites of the enzyme to produce catalytic and regulatory fragments. The solubilized 46-kDa fragment was identical with the catalytic fragment of beta II-PKC. The results indicate that this PKC subspecies is tightly associated with the cytoskeletal network of synaptic membranes.


ENZYMATIC ENHANCEMENT OF INFECTIVITY OF REOVIRUS.
Spendlove, R. S. (California State Department of Public Health, Berkeley), and F. L. Schaffer. Enzymatic enhancement of infectivity of reovirus. J. Bacteriol. 89:597-602. 1965.-Enhancement of infectivity by chymotrypsin treatment has been demonstrated with all three types of reovirus, although not in all viral preparations. Enzyme treatment did not produce a simultaneous increase in the hemagglutinating activity of reovirus type 1 (the only type tested). The infectivity of reovirus type 1 (Lang strain) was increased by treatment with chymotrypsin, trypsin, papain, and a filtrate from a culture of a Pseudomonas sp. but not by treatment with pepsin. Sedimentation experiments showed that the property of enhanceability was closely associated with the virus particles themselves. Results of studies involving various sequential treatments with chymotrypsin, and with heat in the presence or absence of 2 m MgCl(2), were compatible with the interpretation that inhibited virus is resistant to exposure to a temperature of 56 C in the absence of MgCl(2), whereas activated virus is thermolabile. Activation of reovirus infectivity by heat in the presence of MgCl(2) and by chymotrypsin was not additive.


How Would You Manage This Patient With Heart Failure With Preserved Ejection Fraction? : Grand Rounds Discussion From Beth Israel Deaconess Medical Center.
The proportion of patients with new-onset heart failure who have preserved rather than reduced left ventricular ejection fraction (HFpEF and HFrEF) has been increasing over recent decades. In fact, HFpEF now outweighs HFrEF as the predominant heart failure subtype and likely remains underdiagnosed in the community. This is due in part to an aging population and a rise in other risk factors for HFpEF, including obesity and associated cardiometabolic disease. Whereas the diagnosis of HFrEF is relatively straightforward, the diagnosis of HFpEF is often more challenging because there can be other causes for symptoms, including dyspnea and fatigue, and cardinal physical examination findings of elevated jugular venous pressure or pulmonary congestion may not be evident at rest. In 2022, the American College of Cardiology, the American Heart Association, and the Heart Failure Society of America published a comprehensive guideline on heart failure that included recommendations for the management of HFpEF. The use of diuretics for the management of congestion remained the only class 1 (strong) recommendation. New recommendations included broader use of sodium-glucose cotransporter-2 inhibitors (SGLT2i, class 2a), and angiotensin receptor-neprilysin inhibitors (class 2b). In 2023, the American College of Cardiology published an expert consensus decision pathway for the management of HFpEF that suggests treatment strategies based on sex assigned at birth, ejection fraction, clinical evidence of congestion, and candidacy for SGLT2i therapy. Here, 2 experts, a cardiologist and a geriatrician, discuss their approach to the diagnosis and management of HFpEF and how they would apply guidelines to an individual patient.


S-Glutathionylation and S-Nitrosylation in Mitochondria: Focus on Homeostasis and Neurodegenerative Diseases.
Redox post-translational modifications are derived from fluctuations in the redox potential and modulate protein function, localization, activity and structure. Amongst the oxidative reversible modifications, the S-glutathionylation of proteins was the first to be characterized as a post-translational modification, which primarily protects proteins from irreversible oxidation. However, a growing body of evidence suggests that S-glutathionylation plays a key role in core cell processes, particularly in mitochondria, which are the main source of reactive oxygen species. S-nitrosylation, another post-translational modification, was identified >150 years ago, but it was re-introduced as a prototype cell-signaling mechanism only recently, one that tightly regulates core processes within the cell's sub-compartments, especially in mitochondria. S-glutathionylation and S-nitrosylation are modulated by fluctuations in reactive oxygen and nitrogen species and, in turn, orchestrate mitochondrial bioenergetics machinery, morphology, nutrients metabolism and apoptosis. In many neurodegenerative disorders, mitochondria dysfunction and oxidative/nitrosative stresses trigger or exacerbate their pathologies. Despite the substantial amount of research for most of these disorders, there are no successful treatments, while antioxidant supplementation failed in the majority of clinical trials. Herein, we discuss how S-glutathionylation and S-nitrosylation interfere in mitochondrial homeostasis and how the deregulation of these modifications is associated with Alzheimer's, Parkinson's, amyotrophic lateral sclerosis and Friedreich's ataxia.


Mining Whole-liver Information with Deep Learning for Preoperatively Predicting HCC Recurrence-free Survival.
Hepatocellular carcinoma (HCC) is globally a leading cause of cancer death. Non-invasive pre-operative prediction of HCC recurrence-free survival (RFS) after resection is essential but remains challenging. Previous models based on medical imaging focus only on tumor area while neglecting the whole liver condition. In fact, HCC patients usually suffer from chronic liver diseases which also hamper the patient survival. This work aims to develop a novel convolutional neural network (CNN) to mine whole-liver information from contrast-enhanced computed tomography (CECT) to predict RFS after hepatic resection in HCC. Our proposed RFSNet takes liver regions from CECT as input, and outputs a risk score for each patient. Cox proportional-hazards loss was applied for model training. A total of 215 patients with primary HCC and treated with hepatic resection were included for analysis. Patients were randomly split into developing subcohort and testing subcohort by 4:1. The developing subcohort was further split into the training subcohort and validation subcohort for model training. Baseline models were built with tumor region, radiomics features and/or clinical features the same as previous tumor-based approaches. Results showed that RFSNet achieved the best performance with concordance-indinces (CIs) of 0.88 and 0.65 for the developing and testing subcohorts, respectively. Adding clinical features did not improve RFSNet. Our findings suggest that the proposed RFSNet based on whole liver is able to extract more valuable information concerning RFS prognosis compared to features from only tumor and the clinical indicators.


Utility of radiolabeled somatostatin receptor analogues for staging/restaging and treatment of somatostatin receptor-positive pediatric tumors.
In this paper, we review the pediatric oncologic applications of somatostatin receptor-targeted imaging and therapy. Somatostatin receptors are expressed in high densities by embryonal tumors, such as neuroblastoma and medulloblastoma, and neuroendocrine tumors like carcinoids and islet cell tumors. We first review the distribution of these receptors in normal tissues and tumor cells. We then discuss the technique of somatostatin receptor scintigraphy (SRS) in the pediatric population. Next, the specific clinical applications of SRS with regard to the imaging of neuroblastoma, central nervous system tumors, and gastroenteropancreatic neuroendocrine tumors of childhood are discussed. Finally, we discuss the potential role of somatostatin receptor-targeted radiotherapy for improving the duration and quality of life of children with these tumors.


Differential docking of high-affinity peptide ligands to type A and B cholecystokinin receptors demonstrated by photoaffinity labeling.
Type A and B cholecystokinin (CCK) receptors are highly homologous members of the class-I family of G protein-coupled receptors that bind CCK with high affinity. However, they have divergent structural specificities, with the type A receptor requiring seven carboxyl-terminal residues including a sulfated tyrosine and the type B receptor requiring only the carboxyl-terminal tetrapeptide. The aim of this work was to utilize affinity labeling to determine spatial approximations with photolabile p-benzoyl-l-phenylalanine (Bpa) residues sited at each end of CCK as docked at the type B CCK receptor, contrasting this with analogous work using similar probes docked at the type A receptor. Both probes were fully efficacious, potent agonists that stimulated intracellular calcium in receptor-bearing CHO-CCKBR cells (EC(50) values: Bpa(24) probe, 41 +/- 9 pM; Bpa(33) probe, 15 +/- 3.3 pM). They bound specifically, with high affinity (K(i) values: Bpa(24) probe, 0.60 +/- 0.17 nM; Bpa(33) probe, 0.58 +/- 0.11 nM). Cyanogen bromide cleavage of the covalently labeled receptor suggested the first extracellular loop as the region of labeling by each probe, distinct from the type A CCK receptor regions labeled using the same probes (third loop and amino-terminal tail, respectively). This was confirmed by subsequent enzymatic and chemical cleavage of labeled wild-type and mutant receptors. Sequential cycles of Edman degradation of labeled receptor fragments identified the specific residues within loop one labeled by each probe (Bpa(24) probe labeled Phe(122); Bpa(33) probe labeled Thr(119)). This provides a direct demonstration of distinct modes of docking the same high-affinity ligand to highly homologous receptors.


Interrogation and validation of the interactome of neuronal Munc18-interacting Mint proteins with AlphaFold2.
Munc18-interacting proteins (Mints) are multi-domain adaptors that regulate neuronal membrane trafficking, signalling and neurotransmission. Mint1 and Mint2 are highly expressed in the brain with overlapping roles in the regulation of synaptic vesicle fusion required for neurotransmitter release by interacting with the essential synaptic protein Munc18-1. Here, we have used AlphaFold2 to identify and then validate the mechanisms that underpin both the specific interactions of neuronal Mint proteins with Munc18-1 as well as their wider interactome. We find a short acidic α-helical motif (AHM) within Mint1 and Mint2 is necessary and sufficient for specific binding to Munc18-1 and binds a conserved surface on Munc18-1 domain3b. In Munc18-1/2 double knockout neurosecretory cells mutation of the Mint-binding site reduces the ability of Munc18-1 to rescue exocytosis, and although Munc18-1 can interact with Mint and Sx1a proteins simultaneously in vitro we find they have mutually reduced affinities, suggesting an allosteric coupling between the proteins. Using AlphaFold2 to then examine the entire cellular network of putative Mint interactors provides a structural model for their assembly with a variety of known and novel regulatory and cargo proteins including ARF3/ARF4 small GTPases, and the AP3 clathrin adaptor complex. Validation of Mint1 interaction with a new predicted binder TJAP1 provides experimental support that AlphaFold2 can correctly predict interactions across such large-scale datasets. Overall, our data provides insights into the diversity of interactions mediated by the Mint family and shows that Mints may help facilitate a key trigger point in SNARE complex assembly and vesicle fusion.


Fumarate hydratase inactivation in hereditary leiomyomatosis and renal cell cancer is synthetic lethal with ferroptosis induction.
Hereditary leiomyomatosis and renal cell cancer (HLRCC) is a hereditary cancer syndrome characterized by inactivation of the Krebs cycle enzyme fumarate hydratase (FH). HLRCC patients are at high risk of developing kidney cancer of type 2 papillary morphology that is refractory to current radiotherapy, immunotherapy and chemotherapy. Hence, an effective therapy for this deadly form of cancer is urgently needed. Here, we show that FH inactivation (FH-/- ) proves synthetic lethal with inducers of ferroptosis, an iron-dependent and nonapoptotic form of cell death. Specifically, we identified gene signatures for compound sensitivities based on drug responses for 9 different drug classes against the NCI-60 cell lines. These signatures predicted that ferroptosis inducers would be selectively toxic to FH-/- cell line UOK262. Preferential cell death against UOK262-FH-/- was confirmed with 4 different ferroptosis inducers. Mechanistically, the FH-/- sensitivity to ferroptosis is attributed to dysfunctional GPX4, the primary cellular defender against ferroptosis. We identified that C93 of GPX4 is readily post-translationally modified by fumarates that accumulate in conditions of FH-/- , and that C93 modification represses GPX4 activity. Induction of ferroptosis in FH-inactivated tumors represents an opportunity for synthetic lethality in cancer.


Double asymmetric hydrogenation of conjugated dienes: a self-breeding chirality route for C2 symmetric 1,4-diols.
The synthesis and double asymmetric hydrogenation of (Z,Z)-1,3-diene-1,4-diyl diacetates is described. In this reaction C2/meso ratios up to 85 : 15 and enantioselectivities up to 97% ee have been achieved. As the hydrogenation products can be converted into chiral 1,4-diols, key starting materials for the preparation of the best catalysts used, this catalytic system enables a self-breeding chirality process.


A hot-deck multiple imputation procedure for gaps in longitudinal recurrent event histories.
We propose a regression-based hot-deck multiple imputation method for gaps of missing data in longitudinal studies, where subjects experience a recurrent event process and a terminal event. Examples are repeated asthma episodes and death, or menstrual periods and menopause, as in our motivating application. Research interest concerns the onset time of a marker event, defined by the recurrent event process, or the duration from this marker event to the final event. Gaps in the recorded event history make it difficult to determine the onset time of the marker event, and hence, the duration from onset to the final event. Simple approaches such as jumping gap times or dropping cases with gaps have obvious limitations. We propose a procedure for imputing information in the gaps by substituting information in the gap from a matched individual with a completely recorded history in the corresponding interval. Predictive mean matching is used to incorporate information on longitudinal characteristics of the repeated process and the final event time. Multiple imputation is used to propagate imputation uncertainty. The procedure is applied to an important data set for assessing the timing and duration of the menopausal transition. The performance of the proposed method is assessed by a simulation study.


Elucidation of the physicochemical properties and potency of siRNA-loaded small-sized lipid nanoparticles for siRNA delivery.
Because nanoparticles with diameters less than 50nm penetrate stromal-rich tumor tissues more efficiently, the synthesis of small-sized nanoparticles encapsulating short interfering RNA (siRNA) is important in terms of realizing novel siRNA medicine for the treatment of various cancers. Lipid nanoparticles (LNPs) are the leading systems for the delivery of siRNA in vivo. Limit size LNPs were successfully synthesized using a microfluidic mixing technique. However, the physicochemical properties and potential for in vivo siRNA delivery of the limit-size LNPs have not been examined in detail. In the present study, we prepared LNPs with different diameters from 32 to 67nm using a microfluidic mixing devise and examined the physicochemical properties of the particles and the potential for their use in delivering siRNA in vitro and in vivo to liver. Reducing the size of the LNPs causes poor-packing and an increased surface area, which result in their instability in serum. Moreover, it was revealed that the ability of endosomal escape (cytosolic siRNA release) of the smaller LNPs is subject to inhibition by serum compared to that of larger counterparts. Taken together, an increase in packing and avoiding the adsorption of serum components are key strategies for the development of next-generation highly potent and small-sized LNPs.


Social phobia: diagnosis and epidemiology, neurobiology and pharmacology, comorbidity and treatment.
Social phobia is a common disorder associated with significant psychosocial impairment, representing a substantial public health problem largely determined by the high prevalence, and the lifelong chronicity. Social phobia starts in early childhood or adolescence and is often comorbid with depression, other anxiety disorders, alcohol and substance abuse or eating disorders. This cascade of comorbidity, usually secondary to social phobia, increases the disability associated with the condition. The possibility that social phobia may be a trigger for later developing comorbid disorders directs attention to the need for early effective treatment as a preventive measure. The most recent drug class to be investigated for the psychopharmacological treatment of social phobia is the SSRI group for which there is growing support. The other drug classes that have been evaluated are monoamine oxidase inhibitors (MAOIs), benzodiazepines, and beta-blockers. The SSRIs represent a new and attractive therapeutic choice for patients with generalized social phobia. Recently the first, large scale, placebo-controlled study to assess the efficacy of drug treatment in generalized social phobia has been completed with paroxetine. Paroxetine was more effective in reducing the symptoms than placebo and was well tolerated. Many now regard SSRIs as the drugs of choice in social phobia because of their effectiveness and because they avoid the problems of treatment with benzodiazepines or classical MAOIs.


Enhancement of lobster growth.
Selected captive lobsters have been successfully mated at the Massachusetts Lobster Hatchery. Progeny raised in warm seawater grew at least four times as fast as lobsters grown at ambient ocean temperatures. These studies demonstrate that lobsters will reach sexual maturity (a weight of about 454 grams) in less than 2 years, compared to 8 years at ambient temperatures in Canadian waters. A further reduction in this time has been achieved by phenotypic selection for fast-growing lobsters. Our initial success in accelerating growth and in mating selected parents suggests that lobster farming may be possible.


Multimodal data acquisition at SARS-CoV-2 drive through screening centers: Setup description and experiences in Saarland, Germany.
SARS-CoV-2 drive through screening centers (DTSC) have been implemented worldwide as a fast and secure way of mass screening. We use DTSCs as a platform for the acquisition of multimodal datasets that are needed for the development of remote screening methods. Our acquisition setup consists of an array of thermal, infrared and RGB cameras as well as microphones and we apply methods from computer vision and computer audition for the contactless estimation of physiological parameters. We have recorded a multimodal dataset of DTSC participants in Germany for the development of remote screening methods and symptom identification. Acquisition in the early stages of a pandemic and in regions with high infection rates can facilitate and speed up the identification of infection specific symptoms and large-scale data acquisition at DTSC is possible without disturbing the flow of operation.


Temporal and spatial variation in major ion chemistry and source identification of secondary inorganic aerosols in Northern Zhejiang Province, China.
To investigate the seasonal and spatial variations of ion chemistry of fine particles in Northern Zhejiang Province (NZP), China, one year-long field sampling was conducted at four representative sites (two urban, one suburb, and one rural sites) in both cities of Hangzhou and Ningbo from December 2014 to November 2015. Twelve water soluble inorganic ions (WSII) were characterized in this comprehensive study. The annual average of PM2.5 concentration in NZP as overall was 66.2 ± 37.7 μg m-3, and urban sites in NZP were observed with more severe PM2.5 pollution than the suburban and rural sites. The annual average concentration of total WSII at four sampling sites in NZP was 29.1 ± 19.9 μg m-3, dominated by SO42- (10.3 μg m-3), and followed by NO3- (8.9 μg m-3), NH4+ (6.6 μg m-3), Cl- (1.3 μg m-3) and K+ (0.7 μg m-3). Among all cations, NH4+ was the predominant neutralizing ion with the highest neutralization factor (NF), while the remaining cations showed limited neutralization capacity. The highest and lowest sulfur oxidation ratio (SOR) values in this region were found in summer and winter, respectively; while the seasonal patterns for nitrogen oxidation ratio (NOR) were opposite to that of SOR. Principal component analysis (PCA) showed that the significant sources of WSII in NZP were industrial emissions, biomass burning, and formation of secondary inorganic aerosols. In addition, contribution from transboundary transport of polluted aerosols was also confirmed from the assessment through air mass backward trajectory analysis.


UV-promoted radical formation, and near-IR-induced and spontaneous conformational isomerization in monomeric 9-methylguanine isolated in low-temperature Ar matrices.
Three low-energy isomers of 9-methylguanine, the amino-oxo (AO) form and two amino-hydroxy (AH1 and AH2) conformers, were trapped from the gas phase into low-temperature argon matrices. The AH1 and AH2 isomers, differing in the orientation of the OH group, were found to transform into each other upon excitation with near-IR light. The population of the AO form of the compound was not changed upon any near-IR irradiation of the matrix samples. Using monochromatic near-IR light, generated by a frequency-tunable laser source, it was possible to selectively induce the AH1 → AH2 or AH2 → AH1 conversion. Photoreversibility of this conformational transformation was then demonstrated. Exposure of matrix-isolated monomers of 9-methylguanine to broadband near-IR light also led to conformational conversions within the amino-hydroxy tautomeric form; the final stage of this process was always the same photostationary state independent of the initial ratio of AH1 and AH2 populations. Spontaneous conformational conversion, transforming the higher-energy AH2 form into the lower-energy AH1 isomer, was observed for matrix-isolated monomers of 9-methylguanine kept in the dark. The mechanism of this process must rely on quantum tunneling of the light hydrogen atom. Irradiation of matrix-isolated 9-methylguanine with UV laser light at λ = 288 or 285 nm led to a substantial consumption of the two AH forms, while the amount of AO isomer remained unchanged. On the other hand, a decrease in the population of the AO isomer occurred upon excitations at shorter wavelengths, λ = 280 or 275 nm. The spectral changes observed after UV-irradiation suggest the generation (and stabilization in the matrix) of a radical species, resulting from the photocleavage of the O-H or N1-H bonds, in the AH or AO isomer, respectively.


Are Myths and Preconceptions Preventing us from Applying Ionic Liquid Forms of Antiviral Medicines to the Current Health Crisis?
At the moment, there are no U.S. Food and Drug Administration (U.S. FDA)-approved drugs for the treatment of COVID-19, although several antiviral drugs are available for repurposing. Many of these drugs suffer from polymorphic transformations with changes in the drug's safety and efficacy; many are poorly soluble, poorly bioavailable drugs. Current tools to reformulate antiviral APIs into safer and more bioavailable forms include pharmaceutical salts and cocrystals, even though it is difficult to classify solid forms into these regulatory-wise mutually exclusive categories. Pure liquid salt forms of APIs, ionic liquids that incorporate APIs into their structures (API-ILs) present all the advantages that salt forms provide from a pharmaceutical standpoint, without being subject to solid-state matter problems. In this perspective article, the myths and the most voiced concerns holding back implementation of API-ILs are examined, and two case studies of API-ILs antivirals (the amphoteric acyclovir and GSK2838232) are presented in detail, with a focus on drug property improvement. We advocate that the industry should consider the advantages of API-ILs which could be the genesis of disruptive innovation and believe that in order for the industry to grow and develop, the industry should be comfortable with a certain element of risk because progress often only comes from trying something different.


Congruence between position descriptions for public health nursing directors and supervisors with national professional standards and competencies.
This study described the extent to which position descriptions (PDs) for public health directors of nursing (DONs) and non-DON public health nursing (PHN) supervisors in Ohio local health departments incorporate national standards of PHN practice and competencies for public health managers. Ninety-four PDs were obtained from 66 local health departments. Statements in each PD were analyzed for congruence with the 2007 American Nurses Association Public Health Nursing Scope and Standards of Practice (ANA Standards) and the Council on Linkages Core Competencies for Public Health Professionals (COL Competencies). Health department and PD characteristics were also obtained. COL Competencies within each COL skill domain were pooled for analysis. Position descriptions addressed an average of 7.6 of the 20 ANA Standards/Substandards (range, 1-15). The most commonly addressed ANA Standards were Leadership, Regulatory Activities, Collegiality and Professional Relationships, and Planning; the least often addressed were Population Diagnosis and Priorities, Professional Practice Evaluation, Outcomes Identification, Advocacy, and Evaluation. Position descriptions addressed an average of 3.6 of the 8 COL skill domains (range, 0-6). Financial Planning and Management, Policy Development/Program Planning, Community Dimensions of Practice, and Analytic/Assessment were the most commonly addressed COL skill domains, whereas Cultural Competence and Basic Public Health Sciences were the least commonly addressed. About 75% of the PDs included task statements that did not correspond to any of the ANA Standards or COL Competencies. Results indicate that PDs do not reflect compliance with professional mandates for the practice of PHN. This lack of fit between PDs and nationally recognized standards of practice and competencies suggest that PHN may be undifferentiated as a public health discipline and as a nursing specialty.


Are there any differences among psoriasis, psoriatic arthritis and rheumatoid arthritis in terms of metabolic syndrome and cardiovascular risk factors?
Although the frequency of metabolic syndrome has been studied separately in psoriasis, psoriatic arthritis (PsA), and rheumatoid arthritis (RA) patients, there is no study that compares the prevalence of metabolic syndrome in all three diseases. The purpose of this study is to evaluate the relationship between metabolic syndrome (MetS) and chronic low-grade inflammatory diseases, and to determine the frequency of MetS and insulin resistance in psoriasis and PsA as compared to RA. A total of 155 patients were included in this cross-sectional study. Fifty patients who were diagnosed with psoriasis, 55 PsA patients who were diagnosed according to the CASPAR criteria, and 50 seropositive RA patients who were diagnosed according to the ACR/EULAR 2010 classification criteria were included in this study. MetS was diagnosed by the 2005 criteria of International Diabetes Federation. The cardiovascular risk factors and parameters associated with MetS were evaluated. The patients' mean age was significantly higher in the RA. MetS was determined in 33.5% of all patients and MetS and insulin resistance showed no significant difference among the three groups (psoriasis: 36%, PsA: 29%, RA: 36%; p: 0.684 and psoriasis: 70%, PsA: 64%, RA: 66%, respectively; p: 0.785). Triglyceride levels were higher in psoriasis and PsA as compared to the RA (psoriasis: 34%, PsA: 32.7%, RA: 16%, respectively; p: 0.045). The frequency of hypertension was 38% in the RA, which was higher than PsA and psoriasis (p: 0.011). In all three groups, the prevalence of MetS was shown to be higher than the general population. The lack of difference between these groups may be due to the small number of patients, the retrospective study design, and the inequality of the population with respect to age and gender.


Atherogenic lipoproteins enhance mesangial cell expression of platelet-derived growth factor: role of protein tyrosine kinase and cyclic AMP-dependent protein kinase A.
Mesangial cell proliferation and extracellular matrix accumulation are fundamental in the pathogenesis of glomerulosclerosis. Platelet-derived growth factor (PDGF) is a major cytokine involved in mesangial cell proliferation, and its increased expression is seen in glomerular injury. Atherogenic lipoproteins stimulate mesangial cell proliferation and induce glomerular injury in experimental animals. We examined the effect of low-density lipoprotein (LDL) and its more atherogenic oxidized forms, minimally modified LDL (mm-LDL) and oxidized LDL (ox-LDL) on mesangial cell PDGF mRNA expression. Incubation with 2.5 to 25 microg/ml LDL or mm-LDL for 1 to 4 hours stimulated mesangial cell PDGF mRNA expression (mm-LDL 2 to 3 times greater than LDL); ox-LDL had no effect. Similarly, both LDL and mm-LDL induced mesangial cell DNA synthesis (mm-LDL 1.5 to 2 times greater). In further studies evaluating key associated intracellular signal transduction mechanisms, the protein tyrosine kinase (PTK) inhibitors herbimycin and genistein markedly decreased basal and lipoprotein-induced PDGF mRNA expression. Both pertussis toxin and isoproterenol, cyclic AMP-generating substances, stimulated PDGF mRNA expression. Preincubation with H-8 or H-89, cyclic AMP-dependent protein kinase A (PKA) inhibitors, blocked the lipoprotein-induced PDGF message, whereas preincubation with calphostin C, a protein kinase C inhibitor, did not alter LDL- or mm-LDL-mediated PDGF mRNA expression. These data suggest that the accumulation of atherogenic lipoproteins and their endogenous oxidized forms within the glomerulus may regulate mesangial cell PDGF expression and related cellular responses. These events appear to be modulated by signal transduction pathways involving PTK and PKA.


Genotype and subtype profiling of PSI-7977 as a nucleotide inhibitor of hepatitis C virus.
PSI-7977, a prodrug of 2'-F-2'-C-methyluridine monophosphate, is the purified diastereoisomer of PSI-7851 and is currently being investigated in phase 3 clinical trials for the treatment of hepatitis C. In this study, we profiled the activity of PSI-7977 and its ability to select for resistance using a number of different replicon cells. Results showed that PSI-7977 was active against genotype (GT) 1a, 1b, and 2a (strain JFH-1) replicons and chimeric replicons containing GT 2a (strain J6), 2b, and 3a NS5B polymerase. Cross-resistance studies using GT 1b replicons confirmed that the S282T change conferred resistance to PSI-7977. Subsequently, we evaluated the ability of PSI-7977 to select for resistance using GT 1a, 1b, and 2a (JFH-1) replicon cells. S282T was the common mutation selected among all three genotypes, but while it conferred resistance to PSI-7977 in GT 1a and 1b, JFH-1 GT 2a S282T showed only a very modest shift in 50% effective concentration (EC(50)) for PSI-7977. Sequence analysis of the JFH-1 NS5B region indicated that additional amino acid changes were selected both prior to and after the emergence of S282T. These include T179A, M289L, I293L, M434T, and H479P. Residues 179, 289, and 293 are located within the finger and palm domains, while 434 and 479 are located on the surface of the thumb domain. Data from the JFH-1 replicon variants showed that amino acid changes within the finger and palm domains together with S282T were required to confer resistance to PSI-7977, while the mutations on the thumb domain serve to enhance the replication capacity of the S282T replicons.


Posterior circulation infarcts in patients with vertebrobasilar dolichoectasia.
Vertebrobasilar dolichoectasia (VBD) may produce symptoms by direct compression of cranial nerves or the brain stem, by obstructive hydrocephalus, or by ischemia in the vertebrobasilar arterial territory. This study was undertaken to examine and characterize clinical and imaging findings in patients with stroke associated with VBD and compare these data with those for patients with VBD who did not have a stroke. We studied 40 consecutive stroke patients with associated VBD. All were evaluated by CT scan (n=9), MRI (n=6), or both (n=25). The diameter of the basilar artery (BA), height of bifurcation, and transverse position were evaluated. Clinical and imaging findings were compared with those found in a group of 40 VBD patients without stroke. More than half of the patients (24 of 40) had infratentorial infarcts, located mainly in the midpons. Sixteen patients had supratentorial lesions localized in the thalamus (n=8) or in the superficial arterial territory of the posterior cerebral artery (PCA; n=8). The diameter and height of the bifurcation of the BA were correlated with the location of the lesion (PCA territory versus BA territory), in that severe ectasia and vertical elongation of the BA were significantly more often observed in patients with infarcts in PCA territory than in patients with infarcts in territories supplied by branches of the BA. Comparison of VBD patients with and without stroke showed that the incidence of arterial hypertension and the degree of ectasia and lateral displacement of the BA were not significantly different in the two groups. Patients with stroke more often had atherosclerotic changes of the posterior circulation (P=.0006) and a higher degree of vertical elongation of the BA (P=.025). In patients with VBD, superimposed atheromatous changes of the posterior circulation may have an important role in precipitating ischemia. However, other factors related to the severity of the dolichoectasia also favor ischemia and in some cases are the only factors responsible.


Reproducibility and responsiveness of health status measures. Statistics and strategies for evaluation.
Before being introduced to wide use, health status instruments should be evaluated for reliability and validity. Increasingly, they are also tested for responsiveness to important clinical changes. Although standards exist for assessing these properties, confusion and inconsistency arise because multiple statistics are used for the same property; controversy exists over how to measure responsiveness; many statistics are unavailable on common software programs; strategies for measuring these properties vary; and it is often unclear how to define a clinically important change in patient status. Using data from a clinical trial of therapy for back pain, we demonstrate the calculation of several statistics for measuring reproducibility and responsiveness, and demonstrate relationships among them. Simple computational guides for several statistics are provided. We conclude that reproducibility should generally be quantified with the intraclass correlation coefficient rather than the more common Pearson r. Assessing reproducibility by retest at one-to-two week intervals (rather than a shorter interval) may result in more realistic estimates of the variability to be observed among control subjects in a longitudinal study. Instrument responsiveness should be quantified using indicators of effect size, a modified effect size statistic proposed by Guyatt, or the use of receiver operating characteristic (ROC) curves to describe how well various score changes can distinguish improved from unimproved patients.


Exhalation of malondialdehyde from the smoke of high grade cigarettes and effectiveness of filtering it.
It has been found that malondialdehyde penetrates during smoking through filters and exhales with smoke from cigarettes such as: "Pall Mall", "Winston", "Camel", "Lucky Strike", "Kansas", "Mustang", "Ronson", "Chesterfield", "Rally", "Oscar" and "Marlboro". These cigarettes exhale malondialdehyde in a different way, in free form in the amount from 8.5 to 23.5 mg/kg cigarettes on an average of 13.3 mg/kg for the whole group and totally (in free and bounded form) from 9.5 to 26.5 mg/kg (average 16.9 mg/kg). Moreover, it has been found that filters retain this aldehyde in free form on an average of 9.9 mg/kg and totally (in free and bounded form) of 19.0 mg/kg. Without filters these amounts would increase aldehyde contents in smoke by 42.7 and 52.9%, respectively. Low malonalogenic properties of tobacco, high effectiveness of cigarette filters, high degree of tar deposition before inhaling into lungs and slow (6 min and longer) smoking decide about low level of exhalation of malondialdehyde in cigarette smoke.


Evaluation of self-supervised pre-training for automatic infant movement classification using wearable movement sensors.
The recently-developed infant wearable MAIJU provides a means to automatically evaluate infants' motor performance in an objective and scalable manner in out-of-hospital settings. This information could be used for developmental research and to support clinical decision-making, such as detection of developmental problems and guiding of their therapeutic interventions. MAIJU-based analyses rely fully on the classification of infant's posture and movement; it is hence essential to study ways to increase the accuracy of such classifications, aiming to increase the reliability and robustness of the automated analysis. Here, we investigated how self-supervised pre-training improves performance of the classifiers used for analyzing MAIJU recordings, and we studied whether performance of the classifier models is affected by context-selective quality-screening of pre-training data to exclude periods of little infant movement or with missing sensors. Our experiments show that i) pre-training the classifier with unlabeled data leads to a robust accuracy increase of subsequent classification models, and ii) selecting context-relevant pre-training data leads to substantial further improvements in the classifier performance.Clinical relevance- This study showcases that self-supervised learning can be used to increase the accuracy of out-of-hospital evaluation of infants' motor abilities via smart wearables.


Methionine Deprivation Induces a Targetable Vulnerability in Triple-Negative Breast Cancer Cells by Enhancing TRAIL Receptor-2 Expression.
Many neoplasms are vulnerable to methionine deficiency by mechanisms that are poorly understood. Because gene profiling studies have revealed that methionine depletion increases TNF-related apoptosis-inducing ligand receptor-2 (TRAIL-R2) mRNA, we postulated that methionine stress sensitizes breast cancer cells to proapoptotic TRAIL-R2 agonists. Human triple (ER/PR/HER2)-negative breast carcinoma cell lines were cultured in control or methionine-free media. The effects of methionine depletion on TRAIL receptor expression and sensitivity to chemotherapy or a humanized agonistic TRAIL-R2 monoclonal antibody (lexatumumab) were determined. The melanoma-associated antigen MAGED2 was silenced to delineate its functional role in sensitizing TNBC cells to methionine stress. An orthotopic TNBC model was utilized to evaluate the effects of dietary methionine deficiency, lexatumumab, or the combination. Methionine depletion sensitized TNBC cells to lexatumumab-induced caspase activation and apoptosis by increasing TRAIL-R2 mRNA and cell surface expression. MCF-10A cells transformed by oncogenic H-Ras, but not untransformed cells, and matrix-detached TNBC cells were highly sensitive to the combination of lexatumumab and methionine depletion. Proteomics analyses revealed that MAGED2, which has been reported to reduce TRAIL-R2 expression, was suppressed by methionine stress. Silencing MAGED2 recapitulated features of methionine deprivation, including enhanced mRNA and cell surface expression of TRAIL receptors and increased sensitivity to TRAIL receptor agonists. Dietary methionine deprivation enhanced the antitumor effects of lexatumumab in an orthotopic metastatic TNBC model. Methionine depletion exposes a targetable defect in TNBC cells by increasing TRAIL-R2 expression. Our findings provide the foundation for a clinical trial combining dietary methionine restriction and TRAIL-R2 agonists. Clin Cancer Res; 21(12); 2780-91. ©2015 AACR.


Effect of sustained water diuresis on prostaglandin E2 excretion in humans.
The relationship of the renal excretion of prostaglandin E2 (PGE2) to urine flow during a water diuresis was examined using radioimmunoassay (RIA) and gas chromatography-mass spectrometry (GC-MS). Seven normal women fasted overnight were water loaded with 20 ml/kg orally, and each hour for 3 h they drank water to replace the urine volume plus 20 ml. The osmolality of the collected urines ranged from 49 to 1,073 mosmol/kg. Assay of urinary PGE2 concentrations by both RIA and GC-MS gave a correlation coefficient of 0.94. Eight normal women were then studied with a water diuresis sustained for 6 h. The excretion of PGE2 (measured with the validated RIA) increased for the 1st 2-3 h (from 1.8 +/- 0.5 to 25.8 +/- 16.6 pg X kg-1 X min-1), but then fell to base-line level by the 5th h (to 2.9 +/- 0.8 pg X kg-1 X min-1) even though the water diuresis was sustained. The urinary concentration vs. time curves for PGE2 and for the freely diffusible solute urea were compared. PGE2 concentration remained elevated for 3 h before falling (from the 100 pg/ml range to 15 pg/ml) while urea concentration decreased steadily from the 1st h. This finding suggests that the early urinary PGE2 excretion was not a washout phenomenon and is consistent with a transient increase in PGE2 synthesis. We conclude that urinary excretion of PGE2 is not a simple function of urine flow after a water load. There is a transient initial increase in urinary PGE2 excretion at the start of a water load that probably reflects an increase in renal PGE2 synthesis. However, with a sustained water diuresis, PGE2 excretion falls, indicating that an enhanced PGE2 synthesis rate is not required to sustain a water diuresis.


[The occurrence, distribution and toxicity of cyanobacteria in the estuary of Lagoa dos Patos, Rio Grande do Sul].
Several blooms of Microcystis aeruginosa have been observed in the Patos Lagoon estuary during the last fifteen years without a proper investigation of their ecological importance or possible toxicity. The present study has identified and quantified the presence of cyanobacteria in the Patos Lagoon estuary, particularly of M. aeruginosa. During this survey, identification and quantification of the main phytoplankton groups were done in relation to geographical distribution in the estuary. The presence of M. aeruginosa colonies in the estuarine region confirmed their superficial distribution throughout the estuarine waters during twelve months with a maximum of 1, 3.10(6) cells. L-1 in December, 1994 and a minimum of 1, 5.10(5) cells. L-1 in August, 1995 and also confirmed that M. aeruginosa originated from waters in the north of the estuary. The period of the highest cell and colonies densities was coincident with high chlorophyll-a levels in surface waters. Toxicity of M. aeruginosa bloom material was determined by bioassay and concentrations of hepatotoxins microcystins were identified by HPLC-DAD. M. aeruginosa blooms were considered highly toxic, presenting a 24 h-LD50 lower than 100 mg.Kg-1 b.w. and a toxin content higher than 1 microgram.mg-1 d.w. Several microcystin variants were found in the extracts with microcystin-LR predominating.


[The specific compound and biological characteristics of agents of nosocomial pneumonia isolated in hospital surgery departments of Makhachkala].
The nosocomial pneumonia is the most prevailing form of hospital-acquired infection. It is the leading cause of mortality among all forms of hospital-acquired infections. In the departments of resuscitation and intensive therapy nosocomial pneumonia consists more than 25% of all hospital-acquired infections. The analysis of lavages of various components of apparatuses of artificial pulmonary ventilation in the departments of resuscitation and intensive therapy of surgery hospital departments demonstrated that gram-negative micro-flora inoculated more often (72.0%). During last years, this type of micro-flora plays leading part in development of severe forms of nosocomial pneumonia. The gram-negative bacteria consisted up to 38/8% of all isolated cultures. The similar micro-flora was isolated and from tracheobronchial lavages and it confirms the possibility of infection spreading by anesthetic breathing equipment.