Seasonal temperature variability observed at abyssal depths in the Arabian Sea.
The abyssal ocean is generally considered an aseasonal environment decoupled from the variabilities observed at and just below the ocean's surface. Herein, we describe the first in-situ timeseries record of seasonal warming and cooling in the Arabian Sea at a depth of 4000 m. The seasonal cycle was observed over the nearly four-year-long record (from November 2018 to March 2022). The abyssal seasonal temperature cycle also exhibited noticeable interannual variability. We investigate whether or not surface processes influence the near-seabed temperature through deep meridional overturning circulation modulated by the Indian monsoon or by Rossby wave propagation. We also consider if bottom water circulation variability and discharge of the dense Persian Gulf and Red Sea Water may contribute to the observed seasonality.


Childhood Sexual Abuse and Intimate Partner Violence among Women in Methadone Treatment: A Direct or Mediated Relationship?
Women in substance use treatment report rates of childhood sexual abuse and intimate partner violence that far exceed those reported by women in the general population. Previous research with nonrandom samples of women in substance use treatment suggests that there is a statistically significant relationship between childhood sexual abuse and intimate partner violence; however, little is known about the mechanisms of risk between these two public health concerns among this population of women. To address this gap in knowledge and to inform intervention strategies, this study examined direct and mediated relationships between childhood sexual abuse and intimate partner violence risk among a random sample of 416 women in methadone treatment. In addition to high rates of childhood sexual abuse (57.9%), intimate partner violence (lifetime prevalence, 89.7%; 6-month prevalence, 78.4%), posttraumatic stress disorder (PTSD, 28.6%), and global psychological distress (19.5%), findings suggest that the relationship between childhood sexual abuse and intimate partner violence is mediated by mental health problems and that women experiencing PTSD or global psychological distress are 2.7 and 2.4 times more likely to experience intimate partner violence than women without such experiences, respectively. Although not a mediator in this relationship, financial independence reduced women's risk of partner violence by two-thirds. The paper includes discussion of social learning and stress and coping theories to explicate the findings and to inform intervention strategies.


Relatively minor influence of individual characteristics on critical wet-bulb globe temperature (WBGT) limits during light activity in young adults (PSU HEAT Project).
Critical environmental limits are temperature-humidity thresholds above which heat balance cannot be maintained for a given metabolic heat production. This study examined the association between individual characteristics [sex, body surface area (AD), aerobic capacity (V̇o2max), and body mass (mb)] and critical environmental limits in young adults at low metabolic rates. Forty-four (20 M/24 F; 23 ± 4 yr) subjects were exposed to progressive heat stress in an environmental chamber at two low net metabolic rates (Mnet); minimal activity (MinAct; Mnet = ∼160 W) and light ambulation (LightAmb; Mnet = ∼260 W). In two hot-dry (HD; ≤25% rh) environments, ambient water vapor pressure (Pa = 12 or 16 mmHg) was held constant and dry-bulb temperature (Tdb) was systematically increased. In two warm-humid (WH; ≥50% rh) environments, Tdb was held constant at 34°C or 36°C, and Pa was systematically increased. The critical wet-bulb globe temperature (WBGTcrit) was determined for each condition. During MinAct, after entry of Mnet into the forward stepwise linear regression model, no individual characteristics were entered into the model for WH (R2adj = 0.01, P = 0.27) or HD environments (R2adj = -0.01, P = 0.44). During LightAmb, only mb was entered into the model for WH environments (R2adj = 0.44, P < 0.001), whereas only V̇o2max was entered for HD environments (R2adj = 0.22; P = 0.002). These data demonstrate negligible importance of individual characteristics on WBGTcrit during low-intensity nonweight-bearing (MinAct) activity with a modest impact of mb and V̇o2max during weight-bearing (LightAmb) activity in extreme thermal environments.NEW & NOTEWORTHY Our laboratory has recently published a series of papers establishing the upper ambient temperature-humidity thresholds for maintaining heat balance, termed critical environmental limits, in young adults. However, no studies have investigated the relative influence of individual characteristics, such as sex, body size, and aerobic fitness, on those environmental limits. Here, we demonstrate the contributions of sex, body mass, body surface area, and maximal aerobic capacity on critical wet-bulb globe temperature (WBGT) limits in young adults.


An analytical model of enterococci inactivation, grazing, and transport in the surf zone of a marine beach.
An analytical model of enterococci (ENT) concentrations in the surf zone of a long sandy beach is constructed considering the physical processes of dilution by rip currents and alongshore littoral drift, and the biological processes of inactivation and mortality by grazing. The solution is used to construct an expression for the length of shoreline adversely impacted by ENT from a point source. Two non-dimensional parameters are developed whose magnitude can be used to ascertain whether dilution, inactivation, or grazing is the dominant sink for ENT in the surf zone. The model is applied to beaches in southern California, USA. Model input parameters related to physical processes and inactivation are compiled from the literature. Laboratory experiments are conducted to determine grazing mortality rates of ENT (6.5 x 10(-6) s(-1)). Results indicate that at the field sites, between 1000 and 5000 m of shoreline are typically impacted by a continuous point source of ENT. Dilution is the primary cause of decline in ENT concentrations within the surf zone, with inactivation secondary and grazing tertiary. Results recommend strategic positioning of point sources and timing of effluent releases to take advantage of high dilution conditions. Our estimates for grazing mortality rates are within the same order of magnitude as some published inactivation rates, thus we cannot rule out the possibility that grazing is an important sink for ENT, especially in low dilution environments like enclosed bays.


High incidence of fever in patients after biointegral pulmonic valved conduit implantation.
To describe our short- and medium-term outcomes using the BioIntegral pulmonic conduit. Between August 2018 and September 2019, the BioIntegral pulmonic valved conduit was used for right ventricular outflow tract reconstruction in 48 patients. The data were retrospectively retrieved from the patient charts. The median age at surgery was 36 months (interquartile range [IQR] = 18-62 months). The diagnoses were pulmonary atresia-ventricular septal defect in 28 patients, absent pulmonary valve in four patients, truncus arteriosus in six patients, TGA-VSD-PS in five patients, conduit stenosis in three patients, and left venticular outflow tract obstruction requiring a Ross operation in two patients. In the postoperative short-term follow-up, 15 patients out of 48 had a high fever. Of these, 12 patients had concomitantly elevated C-reactive protein levels. There were no patients with blood culture positivity. The median postoperative length of hospital stay was 14 days (IQR = 8-21 days). The overall mortality was 4% in two patients, one died of right ventricular failure and multiple organ failure and one died of pulmonary embolism. The two patients who died were not among the 15 patients with fever. However, four patients with fever underwent balloon angioplasty for conduit restenosis in their medium-term follow-up. There was a high incidence of fever and adverse outcomes in the short-term postoperative follow-up of patients in whom the BioIntegral pulmonic valved conduit was implanted. Caution is advisable in using these conduits until there is convincing evidence about the sterilization and storage standards of these grafts.


Hearing loss in children with Fabry disease.
Hearing loss (HL) is a well-known feature of Fabry disease (FD). Its presence and characteristics have mainly been studied in adult patients, while only limited data are available on the presence and degree of HL in children with FD. This prompted us to study hearing sensitivity in pediatric FD patients. All available audiograms of the Dutch and Norwegian children with FD were retrospectively collected. First, hearing sensitivity was determined by studying hearing thresholds at low, high, and ultra-high frequencies in children with FD and comparing them to zero dB HL, i.e., healthy children. In addition, the presence and type of slight/mild HL (defined as hearing thresholds at low frequencies of 25-40 dB HL) and moderate to severe HL (hearing thresholds >40 dB HL) at first visit were analyzed. If available, follow-up data were used to estimate the natural course of hearing sensitivity and HL in children with FD. One-hundred-thirteen audiograms of 47 children with FD (20 boys, median age at first audiogram 12.0 (range 5.1-18.0) years) were analyzed. At baseline, slight/mild or moderate to severe HL was present in three children (6.4%, 2 boys). Follow-up measurements showed that three additional children developed HL before the age of 18. Of these six children, five had sensorineural HL, most likely caused by FD. Compared to healthy children (zero dB HL), FD children showed increased hearing thresholds at all frequencies (p < 0.01), which was most prominent at ultra-high frequencies (>8 kHz). Hearing sensitivity at these ultra-high frequencies deteriorated in a period of 5 years of follow-up. A minority of children with FD show slight/mild or moderate to severe HL, but their hearing thresholds are poorer than the reference values for normal-hearing children. Clinical trials in FD children should demonstrate whether HL can be prevented or reversed by early treatment and should specifically study ultra-high frequencies.


Influence of chrysin on hepatic marker enzymes and lipid profile against D-galactosamine-induced hepatotoxicity rats.
Chrysin is a flavonoid that exists in nature and is the major component of some traditional medicinal herbs. We investigated the hepatoprotective and antihyperlipidaemic potential of chrysin against D-galactosamine (a single intraperitoneal injection 400 mg/kg BW) induced hepatotoxicity in male albino Wistar rats. D-GalN rats exhibited an increased hepato and nephro toxicity marker activities aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and gamma glutamyl transpeptidase and total bilirubin level while urea, uric acid and creatinine and lipid profile. It also negatively affected the serum total protein, albumin and A/G ratio. Rats treated with chrysin at different concentrations (25, 50 and 100 mg/kg BW) caused a significant improvement in serum protein level, decreased hepato and nephro toxicity markers. It also decreased the levels of very low density lipoprotein cholesterol and low density lipoprotein cholesterol while high density lipoprotein cholesterol significantly increased. It also decreased the levels of total cholesterol, phospholipids, triglycerides, free fatty acids in the plasma and tissues of liver and kidney. The effect of chrysin (25 mg/kg) is comparable with silymarin, a known hepatoprotective drug. Chrysin thus exhibits hepatoprotective and antihyperlipidaemic activity.


Surgical volume reduction and the announcement of triage during the 1st wave of the COVID-19 pandemic in Japan: a cohort study using an interrupted time series analysis.
The coronavirus disease (COVID-19) pandemic has caused unprecedented challenges for surgical staffs to minimize exposure to COVID-19 or save medical resources without harmful patient outcomes, in accordance with the statement of each surgical society. No research has empirically validated declines in surgical volume in Japan, based on the usage of surgical triage. We aimed to identify whether the announcement of surgical priorities by each Japanese surgical society may have affected the surgical volume decline during the 1st wave of this pandemic. We extracted 490,719 available cases of patients aged > 15 years who underwent elective major surgeries between July 1, 2018, and June 30, 2020. After the categorization of surgical specialities, we calculated descriptive statistics to compare the year-over-year trend and conducted an interrupted time series analysis to validate the decline of each surgical procedure. Monthly surgical cases of eight surgical specialities, especially ophthalmology and ear/nose/throat surgeries, decreased from April 2020 and reached a minimum in May 2020. An interrupted time series analysis showed no significant trends in oncological and critical surgeries. Non-critical surgeries showed obvious and statistically significant declines in case volume during the 1st wave of the COVID-19 pandemic according to the statement of each surgical society in Japan.


Design and application of self-healable polymeric films and coatings for smart food packaging.
Smart packaging materials enable active control of parameters that potentially influence the quality of a packaged food product. One type of these that have attracted extensive interest is self-healable films and coatings, which show the elegant, autonomous crack repairing ability upon the presence of appropriate stimuli. They exhibit increased durability and effectively lengthen the usage lifespan of the package. Over the years, extensive efforts have been paid to the design and development of polymeric materials that show self-healing properties; however, till now most of the discussions focus on the design of self-healable hydrogels. Efforts devoted to delineating related advances in the context of polymeric films and coatings are scant, not to mention works reviewing the use of self-healable polymeric materials for smart food packaging. This article fills this gap by offering a review of not only the major strategies for fabrication of self-healable polymeric films and coatings but also the mechanisms of the self-healing process. It is hoped that this article cannot only provide a snapshot of the recent development of self-healable food packaging materials, but insights into the optimization and design of new polymeric films and coatings with self-healing properties can also be gained for future research.


Engineering soluble artificial epidermal growth factor receptor mimics capable of spontaneous in vitro dimerization.
Epidermal growth factor receptor (EGFR) is a clinically validated target for a multitude of human cancers. The receptor is activated upon ligand binding through a critical dimerization step. Dimerization can be replicated in vitro by locally concentrating the receptor kinase domains on the surface of lipid-based vesicles. In this study we investigated the use of coiled coils to induce spontaneous receptor kinase domain dimerization in vitro to form non-membrane-bound artificial receptor mimics in solution. Two engineered forms of EGFR kinase domain fused to coiled coil complementary peptides were designed to self-associate upon mixing. Two fusion protein species (P3-EGFR and P4-EGFR) independently showed the same activity and polymerization profile known to exist with EGFR kinase domains. Upon mixing the two species, coiled coil heterodimers were formed that induced EGFR association to form dimers of the kinase domains. This was accompanied by 11.5-fold increase in the phosphorylation rate indicative of kinase domain activation equivalent to the levels achieved using vesicle localization and mimicking in vivo ligand-induced activation. This study presents a soluble tyrosine kinase receptor mimic capable of spontaneous in vitro activation that can facilitate functional and drug discovery studies for this clinically important receptor class.


New Clones on the Block.
Although immune checkpoint blockade (ICB) has yielded striking clinical responses in subsets of cancer patients, the mechanism of action is still unclear. In a recent issue of Nature Medicine, Yost et al., 2019 report that the T cell clones that dominate the intra-tumoral T cell landscape after ICB are distinct from those prior to treatment, a phenomenon referred to by the authors as "clonal replacement."


Characteristics of lactate metabolism phenotype in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is a highly heterogeneous cancer, and more effective prognostic markers are needed. Lactic acid has been proved to be an important metabolite involved in cancer development, metastasis, and the tumor microenvironment, affecting the prognosis of patients. The role of lactic acid metabolism regulators (LAMRs) in HCC is still unclear. In this study, we analyzed the status of LAMRs, a gene list containing lactate from Molecular Signatures database, in HCC and consensus clustering was performed based on these LAMRs. Cluster B showed higher infiltrations of immune cells, higher TME scores, and a poorer prognosis. We further constructed a risk score based on DEGs using LASSO and COX regression analysis between two clusters, which could effectively predict the prognosis of TCGA-LIHC patients. The GSE14520 cohort confirmed the result. We also examined the correlation of risk scores with clinical characteristics, genetic mutations, drug sensitivity, immune checkpoint inhibitors(ICIs), and immunotherapy. In conclusion, our findings will facilitate a further understanding of the role of partial lactate metabolism related genes in HCC and suggest a new risk score to predict prognosis.


Molecular form and subcellular distribution of acid beta-galactosidase in fibroblasts from patients with GM1 gangliosidosis, Morquio B disease and galactosialidosis.
The molecular form and subcellular distribution of acid beta-galactosidase in cultured fibroblasts from patients with beta-galactosidase deficiency (GM1-gangliosidosis, Morquio B disease and galactosialidosis) were studied, using antibodies against three different forms of the human enzyme: a high-molecular-weight multienzymic complex, a recombinant 84-kDa precursor, and a 64-kDa tryptic product of the precursor. The mature enzyme from normal fibroblasts was immunoprecipitated by the anti-complex and anti-64-kDa protein antibodies, but not by the anti-84-kDa precursor one. immunofluorescence staining of normal fibroblasts revealed the granular (lysosomal) distribution with anti-64-kDa protein antibody and the perinuclear reticular distribution with anti-84-kDa precursor antibody, probably representing the Golgi apparatus. Both patterns were demonstrated in Morquio B disease, but the residual enzyme activity was exclusively due to the mature enzyme. In Type 1 galactosialidosis, most of the expressed enzyme was detected as the precursor form with a perinuclear reticular distribution. In type 2 galactosialidosis, more than half of the enzyme activity was due to the mature form with a lysosomal distribution. Fibroblasts from a patient with GM1 gangliosidosis, expressing no beta-galactosidase mRNA, did not react against either anti-64-kDa protein antibody or anti-84-kDa precursor antibody. The combined use of immunoprecipitation and immunostaining was useful for analysing the pathophysiology of the intracellular processing and transport of the mutant beta-galactosidase.


Determinants of effective treatment coverage for posttraumatic stress disorder: findings from the World Mental Health Surveys.
Posttraumatic stress disorder (PTSD) is associated with significant morbidity, but efficacious pharmacotherapy and psychotherapy are available. Data from the World Mental Health Surveys were used to investigate extent and predictors of treatment coverage for PTSD in high-income countries (HICs) as well as in low- and middle-income countries (LMICs). Seventeen surveys were conducted across 15 countries (9 HICs, 6 LMICs) by the World Health Organization (WHO) World Mental Health Surveys. Of 35,012 respondents, 914 met DSM-IV criteria for 12-month PTSD. Components of treatment coverage analyzed were: (a) any mental health service utilization; (b) adequate pharmacotherapy; (c) adequate psychotherapy; and (d) effective treatment coverage. Regression models investigated predictors of treatment coverage. 12-month PTSD prevalence in trauma exposed individuals was 1.49 (S.E., 0.08). A total of 43.0% (S.E., 2.2) received any mental health services, with fewer receiving adequate pharmacotherapy (13.5%), adequate psychotherapy (17.2%), or effective treatment coverage (14.4%), and with all components of treatment coverage lower in LMICs than HICs. In a multivariable model having insurance (OR = 2.31, 95 CI 1.17, 4.57) and severity of symptoms (OR = .35, 95% CI 0.18, 0.70) were predictive of effective treatment coverage. There is a clear need to improve pharmacotherapy and psychotherapy coverage for PTSD, particularly in those with mild symptoms, and especially in LMICs. Universal health care insurance can be expected to increase effective treatment coverage and therefore improve outcomes.


Telemedical Interventions for Chronic Obstructive Pulmonary Disease Management: Umbrella Review.
Chronic obstructive pulmonary disease (COPD) is a growing epidemic, with a heavy associated economic burden. Education, physical activity, and pulmonary rehabilitation programs are important aspects of the management of COPD. These interventions are commonly delivered remotely as part of telemedicine interventions. Several systematic reviews and meta-analyses have been conducted to assess the effectiveness of these interventions. However, these reviews often have conflicting conclusions. We aim to conduct an umbrella review to critically appraise and summarize the available evidence on telemedicine interventions for the management of COPD. In this umbrella review, the MEDLINE, Embase, PsycINFO, and Cochrane databases were searched from inception to May 2022 for systematic reviews and meta-analyses relating to telemedicine interventions for the management of COPD. We compared odds ratios, measures of quality, and heterogeneity across different outcomes. We identified 7 systematic reviews that met the inclusion criteria. Telemedicine interventions used in these reviews were teletreatment, telemonitoring, and telesupport. Telesupport interventions significantly reduced the number of inpatient days and quality of life. Telemonitoring interventions were associated with significant reductions in respiratory exacerbations and hospitalization rates. Teletreatment showed significant effectiveness in reducing respiratory exacerbations, hospitalization rate, compliance (acceptance and dropout rate), and physical activity. Among studies that used integrated telemedicine interventions, there was a significant improvement in physical activity. Telemedicine interventions showed noninferiority or superiority over the standard of care for the management of COPD. Telemedicine interventions should be considered as a supplement to usual methods of care for the outpatient management of COPD, with the aim of reducing the burden on health care systems.


Effects of thoracic and fine PM and their components on heart rate and pulmonary function in COPD patients.
Population-based personal exposures to particulate matter (PM) and personal-ambient relationships of PM and component concentrations for outpatients with COPD and/or asthma were investigated in New York City (NYC) and Seattle for thoracic PM (PM(10)) and fine PM (PM(2.5)). Measurements of outdoor, indoor, and personal PM(10) and PM(2.5) concentrations were made concurrently for 12-consecutive days at 24 patients' residences. Filters were analyzed for elemental components, using XRF and black carbon (BC), by reflectance. Daily morning and evening measurements of heart rate (HR) and blood oxygen saturation (SpO(2)) by pulse oximeter, and forced expiratory volume in 1 s (FEV(1)) and peak expiratory flowrate (PEF) by spirometry were also measured, and symptom data were collected. Central monitoring site, outdoor, indoor, and personal concentration-response relationships of PM(2.5), PM(10-2.5), and their components were examined using mixed-effect models. The relatively small sample size of the study limited the interpretation of results, but of the PM chemical components examined, only nickel concentrations showed consistent associations, and only with HR in the NYC COPD patients.


Effects of substituent pattern on the intracellular target of antiproliferative benzo[b]thiophenyl chromone derivatives.
A new biological scaffold was produced by replacing the 6π-electron phenyl ring-B of a natural flavone skeleton with a 10π-electron benzothiophene (BT). Since aromatic rings are important for ligand protein interactions, this expansion of the π-electron system of ring-B might change the bioactivity profile. One of the resulting novel natural product-inspired compounds, 2-(benzo[b]thiophen-3-yl)-5-hydroxy-7-isopropoxy-6-methoxyflavone (6), effectively arrested the cell cycle at the G2/M phase and displayed significant antiproliferative effects with IC50 values of 0.05-0.08 μM against multiple human tumor cell lines, including a multidrug resistant line. A structure-activity relationship study revealed that a 10π-electron system with high aromaticity, juxtaposed 4-oxo and 5-hydroxy groups, and 7-alkoxy groups were important for potent antimitotic activity. Interestingly, two BT-flavonols (3-hydroxyflavone), 16 and 20, with 3-hydroxy and 5-alkoxy groups, induced distinct biological profiles affecting the cell cycle at the G1/S phase by inhibition of DNA replication through an interaction with topoisomerase I.


A new potentiator of insulin action. Post-receptor activation in vitro.
A new potentiator of insulin action, 5-[4-(1-methylcyclohexylmethoxy)-benzyl]thiazolidine-2,4-dione, was tested for activation of insulin action in vitro. The agent (50 mg X kg-1 X day-1) was orally administered to rats for 14 days and adipocytes from treated rats were used to assess insulin-binding, glucose uptake and glucose oxidation. In obese rats, the agent increased glucose uptake and oxidation without any change in insulin binding, whereas in lean or streptozotocin-treated rats it failed to increase glucose metabolism. Fat tissues were cultured with the agent for 24 h and were tested for insulin action. In the presence of insulin (10 ng/ml) in the culture media, the agent increased glucose oxidation in these cells without any change in insulin binding. However, without insulin in the culture media the agent did not increase glucose oxidation. Thus, the agent appeared to potentiate insulin action at the post receptor process.


Deep generative models for 3D molecular structure.
Deep generative models have gained recent popularity for chemical design. Many of these models have historically operated in 2D space; however, more recently explicit 3D molecular generative models have become of interest, which are the topic of this article. Dozens of published models have been developed in the last few years to generate molecules directly in 3D, outputting both the atom types and coordinates, either in one-shot or adding atoms or fragments step-by-step. These 3D generative models can also be guided by structural information such as a binding pocket representation to successfully generate molecules with docking score ranges similar to known actives, but still showing lower computational efficiency and generation throughput than 1D/2D generative models and sometimes producing unrealistic conformations. We advocate for a unified benchmark of metrics to evaluate generation and propose perspectives to be addressed in next implementations.


Effect of disinfectant solutions on the surface free energy and wettability of filling material.
Gutta-percha (GP) and Resilon (Res) cones are usually disinfected by the use of solutions such as sodium hypochlorite (NaOCl) and chlorhexidine (CHX). The aims of this study were to evaluate the surface free energy of GP and Res cones after disinfection procedures and to investigate the wettability of endodontic sealers in contact with these surfaces. GP and Res flat smooth surfaces were prepared. Thirty-six samples of each material were used. The samples were divided into 6 groups as follows: Control group GP, immersed in Milli-Q water (MQW); CHX group GP, immersed in 2% CHX solution and then washed with MQW; NaOCl group GP, immersed in 5.25% NaOCl and then washed with MQW; Control group Res, immersed in MQW; CHX group Res, immersed in 2% CHX solution and then washed with MQW; and NaOCl group Res, immersed in 5.25% NaOCl and then washed with MQW. Samples were analyzed first with a goniometer to measure the contact angle between the test surfaces and 3 solutions (water, ethylene glycol, and diiodomethane) to determine the surface free energy. Then the contact angle between surfaces and each one of the sealers tested, AH Plus and Real Seal SE, was measured. Data were statistically analyzed. The results showed that when disinfectant solutions were used, the surface free energy was greater. When evaluating the interaction between materials and sealers, the use of CHX presented lower values of contact angle, followed by NaOCl. It was concluded that the solutions used in decontamination increase the surface free energy, promoting high interaction between GP/Res and sealers.


Can the effects of a 1-day CBT psychoeducational workshop on self-confidence be maintained after 2 years? A naturalistic study.
The continued high prevalence of depression in the general population has been in part attributed to a reluctance to consult and also to the limited capacity of psychological therapy services. In a previous randomized controlled trial, self-referral day-long workshops, each for 25 people, offering a cognitive-behavioral therapy approach, seemed to be effective at 3-month follow-up [Brown et al., 2004]. In this study, both experimental group participants and waiting list control participants who went on to attend the workshops (n=102) were followed up and 54.9% provided data after 2 years. The dropout mechanism was investigated and random effects models were used for all analyses. This is a naturalistic study that lacked a control group and had a relatively high attrition rate. The results nevertheless suggest that positive changes in depression, anxiety, distress, and self-esteem achieved at 3 months follow-up were largely maintained at 2 years for those who were "depressed" (Beck Depression Inventory [BDI] scores of 14 and above). However, nondepressed (BDI scores below 14) did not show any significant change. The overall results of this naturalistic study indicate that a very brief, intensive, and large-scale intervention can largely maintain its effects for participants with depression over a 2-year period.


Apoptotic cell nuclei favour aggregation and fluorescence quenching of DNA dyes.
Apoptotic cell nuclei are known to stain hyperchromatically with absorption dyes and dimly with many DNA fluorochromes. We hypothesised that both optical phenomena have the same cause--the ability of apoptotic chromatin to aggregate cationic dyes. This hypothesis was tested using prednisolone-primed rat thymus, which is known to contain apoptotic cells. The apoptotic cells were classified as early and late, based on their morphology, in thin and semithin sections and in thymus imprints on slides. Direct reaction for DNA strand breaks (TUNEL) indicated the presence of breaks in both categories of cells, with more intense labelling in late apoptosis. The chromatin ultrastructure of early apoptotic cells initially retained the supranucleosomal order of packaging which characterises control cells, whereas the dense chromatin of late apoptotic cells possessed the degraded structure. Absorption spectra of the toluidine blue-stained early apoptotic cell chromatin revealed a metachromatic shift, indicating a change of DNA conformation and polymerisation of the dye. When the staining was performed by acridine orange (preceded by a short acid treatment), a paradoxical several-fold increase of fluorescence intensity at a several-fold dilution of the dye was found. The simultaneous reduction of the ratio of red to green components of fluorescence confirmed that the concentration-dependent fluorescence quenching was due to aggregation of the dye. The results suggest that the enhanced affinity of the chromatin of early apoptotic cells for cationic dyes is associated with conformational relaxation rather than degradation of DNA. In late apoptotic cells, the very dense packaging of degraded DNA promotes further aggregation of dyes. The results suggest alternative methods for detection and discrimination of early and late apoptotic cells.


Prediction of Motor Function in Stroke Patients Using Machine Learning Algorithm: Development of Practical Models.
Machine learning (ML) techniques are being increasingly adopted in the medical field. We developed a deep neural network (DNN) model and applied 2 well-known ML algorithms, logistic regression and random forest, in predicting motor outcome at 6 months after stroke. In the present study, by using 14 input variables which are easily measured by clinicians, we developed ML models and investigated their applicability to predicting motor outcome in hemiplegic stroke patients. We retrospectively analyzed data of 1,056 consecutive stroke patients. Favorable outcomes of the upper and lower limbs were defined as a modified Brunnstrom classification (MBC) score of ≥5 (able to perform activities of daily living with the affected upper limb) and a functional ambulation category (FAC) score of ≥4 (able to walk without guardian's assistance), respectively. Poor outcomes of the upper and lower limbs were defined as MBC and FAC scores of <5 and <4, respectively. We developed 3 ML algorithms, namely the DNN, logistic regression, and random forest. Regarding the prediction of upper limb function, for the DNN model, the area under the curve (AUC) was 0.906. For the logistic regression and random forest models, the AUC were 0.874 and 0.882, respectively. For the prediction of lower limb function, for the DNN, logistic regression, and random forest models, the AUCs were 0.822, 0.768, and 0.802, respectively. We demonstrated that the ML algorithms, particularly the DNN, can be useful for predicting motor outcomes in the upper and lower limbs at 6 months after stroke.


A shorter immunoperoxidase technique for the demonstration of carcinoembryonic antigen and other cell products.
A reliable immunoperoxidase schedule for the cellular demonstration of tumour and normal cell products which can be completed in well under two hours is described. The preparations are as clear and readable as those stained by routine histological techniques and should form a valuable adjunct in research and diagnostic studies.


Association of the C(-1019)G 5-HT1A promoter polymorphism with exposure to stressors preceding hospitalization for bipolar depression.
In animals, a higher density of 5-HT1A receptors has been associated with increased behavioral despair after stress. In humans, the G variant of the C(-1019)G 5-HT1A receptor promoter gene polymorphism (rs6295) has been associated with higher expression of 5-HT1A receptors, increased depression, and lower stress preceding completed suicide. We studied the association of rs6295 with the amount of stress in early life and preceding hospitalization for a major depressive episode in course of bipolar disorder. In 74 consecutively admitted inpatients, early life and recent stressors were rated on the Social Readjustment Rating Scale and on the Risky Family Questionnaire. Homozygote carriers of the rs6295 G variant reported less stressful events before current hospitalization for bipolar depression, but not in early life. The G variant was also associated with a higher overall medication load in naturalistic settings before hospitalization. This is the first study that associated 5-HT1A receptor promoter gene variants with stressors preceding the need of hospitalization for bipolar depression. Our findings support the hypothesis that genetic factors affecting serotonergic neurotransmission might contribute to shape the individual resilience to the depressogenic effects of stress in clinical settings.


The normal fundus fluorescein angiogram. V. Intraobserver-, interobserver- and interocular variation of the fundus fluorescein angiogram.
An independent assessment of red free fundus photographs and serial fundus fluorescein angiograms from 11 clinically healthy subjects and 5 diabetics was performed by 3 ophthalmologists twice with an interval of one month and showed no significant interobserver or intraobserver variation (P greater than 0.10). Among 8 clinically healthy subjects, fundus fluorescein angiograms of both eyes disclosed a difference in number and distribution of the angiographic elements (P less than 0.05) between the two eyes in 6 subjects.


Degradation of staphylococcal enterotoxin A by a Pseudomonas aeruginosa isolate from raw milk.
Recently, we found that staphylococcal enterotoxin A (SEA)-producing Staphylococcus aureus strains produced SEA in raw milk with microbial contaminants at high temperatures like 40 °C only. Moreover, the concentration of SEA produced in raw milk gradually decreased after the peak. The reason(s) for SEA degradation in raw milk was studied in this study. Degradation of SEA spiked in raw milk was observed at 40 °C, but not at 25 °C. A Pseudomonas aeruginosa isolate from raw milk degraded SEA spiked in broth at 40 °C. A sample partially purified with a chromatographic method from culture supernatant of the isolate degraded SEA. Two main proteolytic bands were observed in the sample by zymographic analysis with casein. These results suggested that the SEA in raw milk might be degraded by a protease(s) produced by the P. aeruginosa isolate. This finding might be the first report on SEA degradation by a proteolytic enzyme(s) derived from Pseudomonas bacteria to our knowledge.


Evaluating our progress with trauma transfer imaging: repeat CT scans, incomplete imaging, and delayed definitive care.
Many trauma patients present at non-trauma centers and require transfer. CT imaging obtained at the initial hospital (IH) may lead to delays in definitive trauma care, and previous studies have shown imaging is often repeated at the trauma center (TC). A retrospective review was performed of all tier 1 trauma patients transferred to our TC between May 2018 and April 2019. Patients that did and did not undergo CT imaging at the IH were compared (n = 147). Of those with IH CT imaging (n = 68), we identified 4 imaging "inadequacies": (1) repeat CT scans: CT scan of the same body region performed at IH and at TC; (2) C-spine inadequacies: severely injured patients who underwent head CT without a C-spine CT; (3) incomplete chest-abdomen-pelvis (CAP): patients with partial CAP CT imaging at IH that underwent an additional portion of CAP imaging at TC; (4) CAP CT without IV contrast. IH time was significantly prolonged when CT imaging was obtained. Of those that had IH imaging, 13 patients (19%) required repeat CT, ten (15%) had a C-spine inadequacy, 11 (16%) had incomplete CAP, and 28 (41%) had one or more inadequacy. Patients with any inadequacy underwent more CT imaging. Most patients (76%) with imaging at the IH returned to the CT scan at the TC. In severely injured trauma patients transferred to our TC, we identified many continuing issues with IH CT imaging. The imaging inadequacies detailed above lead to delays in definitive care and subject patients to increased radiation.


A biomimetic piezoelectric scaffold with sustained Mg2+ release promotes neurogenic and angiogenic differentiation for enhanced bone regeneration.
Natural bone is a composite tissue made of organic and inorganic components, showing piezoelectricity. Whitlockite (WH), which is a natural magnesium-containing calcium phosphate, has attracted great attention in bone formation recently due to its unique piezoelectric property after sintering treatment and sustained release of magnesium ion (Mg2+). Herein, a composite scaffold (denoted as PWH scaffold) composed of piezoelectric WH (PWH) and poly(ε-caprolactone) (PCL) was 3D printed to meet the physiological demands for the regeneration of neuro-vascularized bone tissue, namely, providing endogenous electric field at the defect site. The sustained release of Mg2+ from the PWH scaffold, displaying multiple biological activities, and thus exhibits a strong synergistic effect with the piezoelectricity on inhibiting osteoclast activation, promoting the neurogenic, angiogenic, and osteogenic differentiation of bone marrow mesenchymal stromal cells (BMSCs) in vitro. In a rat calvarial defect model, this PWH scaffold is remarkably conducive to efficient neo-bone formation with rich neurogenic and angiogenic expressions. Overall, this study presents the first example of biomimetic piezoelectric scaffold with sustained Mg2+ release for promoting the regeneration of neuro-vascularized bone tissue in vivo, which offers new insights for regenerative medicine.


Designing a sustainable logistics network for hazardous medical waste collection a case study in COVID-19 pandemic.
The process of collecting and transporting hazardous medical waste poses a potential threat to the environment and public safety. Furthermore, the waste management system faces higher transportation costs due to the increasing human activities related to rapid population growth. The absence of an efficient and safe logistics network for the timely collection and transportation of hazardous wastes may have negative effects on the environment and public health. Therefore, more sustainable transportation of hazardous waste services is a necessity This paper attempts to design a sustainable network for hazardous medical waste collection services during the COVID-19 pandemic. An electric medical waste collection vehicle routing problem is introduced to construct optimal routes and rosters for a fleet of electric vehicles as well as cover their choice of charging technologies, times and locations. This problem allows us to minimize the health risk of hazardous medical waste while providing cost-effective, zero-emission waste management logistics. Therefore, this problem covers environmental and economic objectives to achieve sustainable development. An effective heuristic that covers adaptive large neighbourhood search and a local search is designed to deal with the complex problem. A series of extensive computational experiments is carried out using real-life benchmark instances to assess the performance of the algorithm. A sensitivity analysis is also conducted to investigate the effect of multiple charger types on the cost and risk objectives. The experiment results indicate that mixed-use of different charger types can reduce the total energy cost and transport risk compared to the case of using only a single charger.


Pro-inflammatory cytokines downregulate platelet derived growth factor-alpha receptor gene expression in human osteoblastic cells.
Platelet derived growth factor (PDGF) is thought to play a significant role in bone repair and regeneration. We previously demonstrated that PDGF-AA binding can be modulated by interleukin-1 (IL-1). We now report that TNF-alpha significantly reduces PDGF-AA binding by decreasing the number of PDGF-alpha receptor subunits on the surface of normal human osteoblastic cells. This inhibition is likely due to a decrease in synthesis of PDGF-alpha receptors since TNF-alpha causes a relatively rapid decrease in PDGF-alpha receptor mRNA levels as determined by Northern blot analysis. The physiologic importance of this inhibition is demonstrated by a TNF-alpha induced decrease in PDGF-AA stimulated tyrosine kinase activity. When saturating concentrations of TNF-alpha were used, the addition of IL-1 further inhibited PDGF-AA binding and further decreased surface expression of PDGF-alpha receptors. In contrast, other mediators such as IL-6, PTH, 1,25(OH)2 vit D3, hydrocortisone, PGE2, bFGF, and IGF-1 had no effect. These results suggest that binding to the PDGF-alpha receptor is decreased by the strong pro-inflammatory cytokines such as IL-1 beta and TNF-alpha rather than as a general response to mediators important in bone resorption or bone formation. TNF-alpha and IL-1 are often co-expressed during destructive inflammatory processes. Thus, TNF-alpha and IL-1 may work in concert to limit the response of osteoblastic cells to PDGF-AA during periods of osseous inflammation.


Clinical and dosimetric predictors of radiation pneumonitis in early-stage lung cancer treated with Stereotactic Ablative radiotherapy (SABR) - An analysis of UK's largest cohort of lung SABR patients.
Stereotactic Ablative Radiotherapy (SABR) is the standard treatment for early-stage medically inoperable lung cancer. Predictors of radiation pneumonitis (RP) in patients treated with SABR are poorly defined. In this study, we investigate clinical and dosimetric parameters, which can predict symptomatic RP in early-stage lung cancer patients treated with SABR. Patients treated with lung SABR between May 2009 and August 2018, in a single United Kingdom (UK) radiotherapy center were included. The patient's baseline characteristics, treatment details, and toxicity were retrieved from the electronic medical record. Dosimetric data was extracted from Xio and Monaco treatment planning systems. Patients were treated according to the UK SABR consortium guidelines. RP was graded retrospectively using Common Terminology Criteria for Adverse Events (CTCAE) version 4.0, based on available clinical and imaging information. Univariate and multivariate binary logistic regression was performed to determine predictive factors for grade ≥ 2 radiation pneumonitis, using Statistical Package for the Social Sciences (SPSS) statistics version 21 software. The goodness of fit was assessed using the Hosmer and Lemeshow test. The optimal diagnostic threshold was tested using the Receiver operating characteristics (ROC) curve. The chi-square test was carried out to test the different risk factors against the likelihood of developing grade ≥ 2 pneumonitis. A total of 1266 patients included in the analysis. The median age of patients was 75 years. Six hundred sixty-six patients (52.6%) were female. Median follow up was 56 months. Sixty-five percent of patients received 55 Gy in 5 fractions. Forty-three percent of patients had Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 2 and 16.2% had PS of 3. The Median Charlson comorbidity index was 6 (range 2-11). Median Standardized Uptake Value (SUV) max of the tumor was 6.5. Four hundred two patients (31.8%) had confirmed histological diagnosis; other patients were treated based on a radiological diagnosis. The median tumor size was 20 mm (range 4 mm-63 mm). Median Planning Target Volume (PTV) was 30.3 cc. Median values of R100, R50, and D2cm were 1.1, 5.6, 32.8 Gy. The median value of mean lung dose, V20, and V12.5 were 3.9 Gy, 5 %and 9.3% respectively. Eighty-five (6.7%) patients developed symptomatic RP (grade ≥ 2) with only 5(0.4%) developing grade 3 RP. Five percent of patients developed rib fractures but only 28% of these were symptomatic. On univariate analysis lower lobe tumor location, larger tumor size, PTV, mean lung dose, lung V20Gy, and V12.5 Gy were significantly associated with grade ≥ 2 RP. On multivariate analysis, only mean lung dose was associated with grade ≥ 2 pneumonitis. ROC curve analysis showed optimal diagnostic threshold for tumour size, PTV, mean lung dose, V20 and V12.5; are 22.5 mm ((Area Under Curve (AUC)-0.565)), 27.15 cc (AUC-0.58), 3.7 Gy (AUC-0.633), 4.6% (AUC-0.597), 9.5% (AUC-0.616). The incidence of ≥grade 2 RP was significantly high for values higher than the ROC threshold. SABR treatment resulted in a very low rate of grade 3 pneumonitis. Lower lobe tumor location, larger tumor size, PTV, mean lung dose, V20, and V12.5 were found to be significant predictors of symptomatic radiation pneumonitis.


Personalized biomarkers to monitor disease progression in advanced non-small-cell lung cancer patients treated with icotinib.
Disease-specific humoral immune response-related protein complexes in blood are associated with disease progression. Thirty-one patients with stage IIIB and IV non-small-cell lung cancer (NSCLC) were administered with oral dose of icotinib hydrochloride (150 mg twice daily or 125 mg 3 times daily) for a 28-continuous-day cycle until diseases progressed or unacceptable toxicity occurred. The levels of immunoinflammation-related protein complexes (IIRPCs) in a series of plasma samples from 31 NSCLC patients treated with icotinib hydrochloride were determined by an optimized native polyacrylamide gel electrophoresis. Three characteristic patterns of the IIRPCs, named as patterns a, b, and c, respectively, were detected in plasma samples from 31 patients. Prior to the treatment, there were 18 patients in pattern a consisting of 5 IIRPCs, 9 in pattern b consisting of six IIRPCs, and 4 in pattern c without the IIRPCs. The levels of the IIRPCs in 27 patients were quantified. Our results indicate that the time length of humoral immune and inflammation response (TLHIIR) was closely associated with disease progression, and the median TLHIIR was 22.0 weeks, 95% confidence interval: 16.2 to 33.0 weeks, with a lead time of median 11 weeks relative to clinical imaging evidence confirmed by computed tomography or magnetic resonance imaging (the median progression-free survival, 34.0 weeks, 95% confidence interval: 27.9 to 49.0 weeks). The complex relationships between humoral immune response, acquired resistance, and disease progression existed. Personalized IIRPCs could be indicators to monitor the disease progression.


A comparison of spatial smoothing methods for small area estimation with sampling weights.
Small area estimation (SAE) is an important endeavor in many fields and is used for resource allocation by both public health and government organizations. Often, complex surveys are carried out within areas, in which case it is common for the data to consist only of the response of interest and an associated sampling weight, reflecting the design. While it is appealing to use spatial smoothing models, and many approaches have been suggested for this endeavor, it is rare for spatial models to incorporate the weighting scheme, leaving the analysis potentially subject to bias. To examine the properties of various approaches to estimation we carry out a simulation study, looking at bias due to both non-response and non-random sampling. We also carry out SAE of smoking prevalence in Washington State, at the zip code level, using data from the 2006 Behavioral Risk Factor Surveillance System. The computation times for the methods we compare are short, and all approaches are implemented in R using currently available packages.


Sedative hypnotics in older people with insomnia: meta-analysis of risks and benefits.
To quantify and compare potential benefits (subjective reports of sleep variables) and risks (adverse events and morning-after psychomotor impairment) of short term treatment with sedative hypnotics in older people with insomnia. Medline, Embase, the Cochrane clinical trials database, PubMed, and PsychLit, 1966 to 2003; bibliographies of published reviews and meta-analyses; manufacturers of newer sedative hypnotics (zaleplon, zolpidem, zopiclone) regarding unpublished studies. Randomised controlled trials of any pharmacological treatment for insomnia for at least five consecutive nights in people aged 60 or over with insomnia and otherwise free of psychiatric or psychological disorders. 24 studies (involving 2417 participants) with extractable data met inclusion and exclusion criteria. Sleep quality improved (effect size 0.14, P < 0.05), total sleep time increased (mean 25.2 minutes, P < 0.001), and the number of night time awakenings decreased (0.63, P < 0.001) with sedative use compared with placebo. Adverse events were more common with sedatives than with placebo: adverse cognitive events were 4.78 times more common (95% confidence interval 1.47 to 15.47, P < 0.01); adverse psychomotor events were 2.61 times more common (1.12 to 6.09, P > 0.05), and reports of daytime fatigue were 3.82 times more common (1.88 to 7.80, P < 0.001) in people using any sedative compared with placebo. Improvements in sleep with sedative use are statistically significant, but the magnitude of effect is small. The increased risk of adverse events is statistically significant and potentially clinically relevant in older people at risk of falls and cognitive impairment. In people over 60, the benefits of these drugs may not justify the increased risk, particularly if the patient has additional risk factors for cognitive or psychomotor adverse events.


Management of symptomatic uterine fibroids after ulipristal acetate suspension and identification of high risk patients for surgery at 6-months of follow-up.
The use of ulipristal acetate (UPA) was indicated for the treatment of uterine fibroids. Following UPA suspension in March 2020, some patients presented worsening and required surgery. We aimed to identify patients at high-risk for undergoing surgery after UPA suspension. We evaluated 85 women receiving intermittent UPA treatment until March 2020. Following UPA suspension, patients received other medical treatments or surgery. The clinico-pathological features were recoded and a quality of life health survey was completed by patients at the time of UPA suspension and at 6-months thereafter. After the suspension of UPA, 17 of the 85 patients receiving intermittent UPA (20%) required surgery, and 68 (80%) required other medical treatments. Patients who underwent surgery were younger and had greater fibroid volume. In our series, 20% of clinically stable patients receiving intermittent UPA required surgery following UPA suspension. These women should be considered for future medical strategies.


Enhanced adjuvanticity of interleukin-2 plasmid DNA administered in polyethylenimine complexes.
We report here the enhanced adjuvant effect of murine interleukin-2 plasmid DNA (mIL-2) by complexation with a cationic polymer polyethylenimine (PEI). The mRNA expression of mIL-2 was observed at C2C12 cells after treatment with pVAXmIL-2 in PEI-complexed form, but not in naked plasmid DNA. In vivo, the plasmid DNA levels of mIL-2 at the administered muscle tissues were prolonged up to 14 days after intramuscular administration in PEI complexes. Moreover, the mRNA expression level in the muscle was 54-fold higher in PEI-complexed pVAXmIL-2 than in naked pVAXmIL-2 at 14 days after intramuscular administration. Mice were immunized on Days 0 and 28 by intramuscular administration of hepatitis B surface antigen (HBsAg) alone or with pVAXmIL-2 in naked or PEI complexes. At 8 weeks after the first immunization, mice coadministered with HBsAg plus pVAXmIL-2 in PEI complexes showed the serum IgG titer 72- and 208-fold higher as compared to those treated with HBsAg plus naked pVAXmIL-2 and with HBsAg alone, respectively. Though both HBsAg-specific IgG(1) and IgG(2a) immune responses were increased upon complexation of pVAXmIL-2 with PEI, IgG(2a) was more effectively induced relative to IgG(1). These results suggest that the complexation with PEI could be useful for prolonging in vivo expression of genetic cytokine adjuvants and enhancing their adjuvanticity for subunit vaccines.


Quaking mouse: an ultrastructural study of the peripheral nerves.
Ultrastructural study of the peripheral nervous system of quaking mice has revealed several unusual features in the pattern of myelination in addition to hypomyelination. These are 1) the presence of 'atypical' Schmidt-Lanterman incisures, 2) irregularities of the nodal and internodal termination of Schwann cell cytoplasm and 3) the presence of non-myelinated segments without associated Schwann cell processes, but covered by basal lamina. In view of the observation of similar features during the development of the P.N.S. and also during P.N.S. remyelination, we suggest that these features are the results of modified myelination due to a defect in the control mechanisms necessary for normal myelination.


Clinical nuclear magnetic resonance imaging of the body.
NMR promises great advances in diagnosis and has delivered so much already that it is expected that in the future it will replace many applications of the currently used imaging modalities. Although x-ray computed tomography is continuing to advance in speed of scanning and resolving power, NMR will most likely soon eliminate its use in many studies of the central nervous system and also in many other areas of the body. The promise of combining topical spectroscopy with imaging is also exciting and should provide further information about metabolic processes of various organs. Progress in NMR is so rapid and the future is so bright that one of the great problems will be to develop a new breed of radiologists who are versatile in biochemistry, mathematics, and computers, as well as competent in morphologic anatomy and pathologic physiology. As time goes on, advances in NMR will be achieved only by teams of clinical and basic scientists encompassing multiple disciplines.


A microassay for Gaucher's disease.
We report a new assay for the detection of individuals heterozygous and homozygous for Gaucher's disease which requires relatively small samples of whole blood (0.3 ml), and which determines 4-methylumbelliferyl-beta-D-glucopyranoside:beta-glucosidase activity under conditions optimal for the determination of leukocyte glucocerebroside:beta-glucocereborsidase activity. The procedure involves the preparation of a leukocyte pellet from 50 mul of whole blood by hypotonic lysis of erythrocytes, followed by assay of beta-glucosidase activity at pH 5.5 in the presence of sodium taurocholate (0.6 g/100 ml). The methods described may also prove to be useful for the diagnosis of other diseases of enzyme deficiency which use fluorogenic substrates and leukocytes as a source of enzyme, such as Fabry's disease, Tay-Sachs disease, and generalized gangliosidosis.


Competitive Effect of Overexpressed C-terminal of Snail-1 (CSnail) in Control of the Growth and Metastasis of Melanoma Cells.
Epithelial-to-mesenchymal transition (EMT) plays a role in the invasion and metastasis of cancer cells. During this phenomenon, Snail can promote tumor progression by upregulating mesenchymal factors and downregulating the expression of pro-apoptotic proteins. Therefore, interventions on the expression rate of Snails may show beneficial therapeutic applications. In this study, the C-terminal region of Snail1, capable of binding to E-box genomic sequences, was subcloned into the pAAV-IRES-EGFP backbone to make complete AAV-CSnail viral particles. B16F10 as a metastatic melanoma cell line, with a null expression of wild type TP53 was transduced by AAV-CSnail. Moreover, the transduced cells were analyzed for in-vitro expression of apoptosis, migration, and EMT-related genes, and in-vivo inhibition of metastasis. In more than 80% of the AAV-CSnail transduced cells, the CSnail gene expression competitively reduced the wild-type Snail functionality and consequently lowered the mRNA expression level of EMT-related genes. Furthermore, the transcription level of cell cycle inhibitory factor p21 and pro-apoptotic factors were promoted. The scratch test showed a decrease in the migration ability of AAV-CSnail transduced group compared to control. Finally, metastasis of cancer cells to lung tissue in the AAV-CSnail-treated B16F10 melanoma mouse model was significantly reduced, pointing out to prevention of EMT by the competitive inhibitory effect of CSnail on Snail1 and increased apoptosis of B16F10 cells. The capability of this successful competition in reducing the growth, invasion, and metastasis of melanoma cells indicates that gene therapy is a promising strategy for the control of the growth and metastasis of cancer cells.


Admission and readmission rate incidences from deprived areas-impact of a classical or multi-dimensional model.
Classical deprivation instruments use a factor analytical approach relying on a smaller number of dimensions, factors or components. Multi-dimensional deprivation models attempt classification in fine detail-even down to street level. Single-centre retrospective cohort study using routinely collected aggregated and anonymised data on emergency medical admissions (96,526 episodes in 50,731 patients; 2002-2016). We calculated admission/readmission rate incidences for the 74 small areas within the hospital catchment area. We compared a classical Small Area Health Research Unit (SAHRU) to the multi-dimensional POBAL Haase and Pratschke Deprivation Index for Small Areas (POBAL) deprivation instrument and their deprivation ranks for two Irish censuses (2006/ 2011). There was poor agreement between the instruments of the Deprivation Ranks by Quintile-with agreement in 46 and 42% of small areas for the respective 2006 and 2011 censuses. The classical model (SAHRU) suggested more areas with severe deprivation (Q5 66 and 55%) compared with POBAL (Q5 32 and 24%) from the respective censuses. SAHRU classical instrument had a higher prediction level incidence rate ratio (IRR) 1.48 (95% CI 1.47, 1.49)) compared with POBAL IRR 1.28 (95% CI 1.27, 1.28) and systematically lower estimates of hospital admission and readmission rate incidences. Earlier Census data modelled more powerfully, suggesting a long latency between social circumstances and the ultimate expression of the emergency medical admission. Deprivation influences hospital incidence rates for emergency medical admissions and readmissions; instruments focusing at the very small area (individual or street level) have a utility but appear inferior in terms of representing the population risk of environmental/socio-economic factors which seem best approximated at a larger scale.


Guiding Opioid Administration by 3 Different Analgesia Nociception Monitoring Indices During General Anesthesia Alters Intraoperative Sufentanil Consumption and Stress Hormone Release: A Randomized Controlled Pilot Study.
This pilot study investigated the effect of sufentanil titration by 3 different analgesia monitoring devices or clinical signs during general anesthesia. Forty-eight patients undergoing radical retropubic prostatectomy with sevoflurane/sufentanil anesthesia were randomly assigned into 4 groups and received sufentanil guided either by 1 of 3 analgesia monitoring devices (Surgical Pleth Index [SPI], Pupillary Pain Index [PPI], Nociception Level [NoL]) or by clinical judgment (control). The primary end point was intraoperative sufentanil consumption. Adrenocorticotropic hormone (ACTH) and cortisol were measured at 4 time points during the day of surgery. Data were analyzed by Kruskal-Wallis and Mann-Whitney U tests and by mixed model and area under the curve (AUC) analyses for group comparisons and time effects of stress hormones. The total amount of sufentanil administration (μg·kg·minute·10) differed between the groups (median [quartiles]: control = 5.6 [4.4-6.4], SPI = 7.2 [4.8-8.4], PPI = 2.0 [1.8-2.9], NoL = 3.8 [3.3-5.1]; PPI versus SPI, -5.1 [-6.6 to -1.3], P < .001; NoL versus SPI, -3.0 [-5.2 to 0.2], P = .024; control versus SPI, -1.6 [-3.7 to 1.7], P = .128; NoL versus PPI, 1.7 [0.6-3.4], P < .001; control versus PPI, 3.4 [2.0-4.6], P < .001; control versus NoL, 1.6 [-0.2 to 3.3], P = .017) (Hodges-Lehmann estimator [99% confidence interval {CI}], P values). The AUC analysis indicated differences among groups in cumulative ACTH levels (ng·liter·minute, natural logarithm (ln)-transformed data) of NoL versus PPI (-1.079 [-1.950 to -0.208], P = .001) and PPI versus SPI (1.192 [0.317-2.068], P= .001), as well as differences in cortisol levels (µg·liter·minute) for PPI versus SPI (46,710 [21,145-72,274], P < .001), NoL versus SPI (27,645 [3163-52,126], P = .003), and control versus SPI (31,824 [6974-56,675], P = .001) (differences in means [99% CI], P value). Secondary end points (postoperative recovery, pain level, and analgesia medication) showed no differences. The type of analgesia nociception monitoring affected the total amount of sufentanil administered. Lower sufentanil doses in the PPI group were associated with an increased endocrine stress response. Titration by SPI caused no opioid reduction compared to the control but was associated with a reduced endocrine stress response.


Canine Zollinger-Ellison syndrome.
The unusual finding of peptic esophagitis and duodenal ulceration in a dog was associated with a malignant pancreatic islet cell tumor producing gastrin and ACTH. The finding of a gastrinoma in a non-human species introduces the potential for developing an animal model for the study of the protean genetic biochemical, physiologic and metabolic aspects of the Zollinger-Ellison syndrome.


Optimal injection method for long-range computed tomography angiography.
We aimed to obtain high-stable computed tomography (CT) values for CT angiography (CTA). We verified the optimal use of the new protocol for long-range scanning using a contrast material flow phantom (Ichikawa, CT imaging theory. Igaku syoin Publishing, Tokyo, 8; Awai et al., Am J Roentgenol 186(2):379-385, 1) to obtain high-stable CT values (Fig. 2). We have developed a novel contrast injection method called the stable line imaging protocol (SLIP). This method involved a gradual reduction in the rate of administration of the contrast medium, which was accompanied by simultaneous administration of saline to compensate for the resultant decrease in the rate of administration. A saline flush was further added after the contrast medium injection. The rate of administration of the saline flush was the same as that of the initial contrast medium. We injected a contrast medium from the injector into the flow phantom. The time-enhancement curve (TEC) data were obtained five times by changing the injection method, and the average CT values were determined. The maximum simulation of the aortic peak enhancement of TEC was defined as pTEC. The time required to keep the CT value more than 80% of the pTEC was defined as the time period of contrast enhancement (p80). The pTEC of a single injection was 295.7 ± 5.3 HU, while that of SLIP was 296.5 ± 2.3 HU; the two values did not differ significantly. The p80 of a single injection was 19.1 ± 0.2 s, while that of SLIP was 23.2 ± 0.4 s; p80 of SLIP was 18% longer than that of the single injection.


An influenza H1 hemagglutinin stem-only immunogen elicits a broadly cross-reactive B cell response in humans.
Current yearly seasonal influenza vaccines primarily induce an antibody response directed against the immunodominant but continually diversifying hemagglutinin (HA) head region. These antibody responses provide protection against the vaccinating strain but little cross-protection against other influenza strains or subtypes. To focus the immune response on subdominant but more conserved epitopes on the HA stem that might protect against a broad range of influenza strains, we developed a stabilized H1 stem immunogen lacking the immunodominant head displayed on a ferritin nanoparticle (H1ssF). Here, we evaluated the B cell response to H1ssF in healthy adults ages 18 to 70 in a phase 1 clinical trial (NCT03814720). We observed both a strong plasmablast response and sustained elicitation of cross-reactive HA stem-specific memory B cells after vaccination with H1ssF in individuals of all ages. The B cell response was focused on two conserved epitopes on the H1 stem, with a highly restricted immunoglobulin repertoire unique to each epitope. On average, two-thirds of the B cell and serological antibody response recognized a central epitope on the H1 stem and exhibited broad neutralization across group 1 influenza virus subtypes. The remaining third recognized an epitope near the viral membrane anchor and was largely limited to H1 strains. Together, we demonstrate that an H1 HA immunogen lacking the immunodominant HA head produces a robust and broadly neutralizing HA stem-directed B cell response.


A transcytotic transport mechanism across the tympanic membrane.
Drug treatments for middle ear diseases are currently delivered systemically, or locally after opening the impermeable tympanic membrane (TM). We previously used bacteriophage display to discover novel peptides that are actively transported across the intact TM, with a variety of transport rates. Peptide structures were analyzed for evidence regarding the mechanism for this unexpected transport, which was then tested by the application of chemical inhibitors. Primary sequences indicated that trans-TM peptides share one of two amino acid motifs. Secondary structures revealed that linear configurations associate with higher transport rates than coiled structures. Tertiary analysis indicated that the shared sequence motifs are prominently displayed at the free ends of rapidly transported peptide phage. The shared motifs were evaluated for similarity to known motifs. The highest probability matches were for protein motifs involved in transmembrane transport and exosomes. Overall, structural findings suggest that the shared motifs represent binding sequences. They also implicate transcytosis, a polarized cell transport mechanism consisting of endocytosis, transcellular transport, and exocytosis. Inhibitor studies indicated that macropinocytosis, retrograde transport through Golgi and exocytosis participate in transport across the TM, consistent with transcytosis. This process can be harnessed to noninvasively deliver therapeutics to the middle ear.


Fine needle aspiration of breast masses. An analysis of 1,398 patients in a community hospital.
To determine the diagnostic efficacy of breast fine needle aspiration (FNA) in the routine practice of a community hospital. The study group consisted of 1,398 consecutive FNA biopsies of the female breast performed at our institution during a five-year period; 450 (32%) had histologic follow-up. Both FNA and the histologic diagnosis were compared in order to establish the accuracy, efficiency and safety of the FNA breast technique. The sensitivity of the FNA procedure was 93.49% and specificity 95.73%, with a predictive value for a positive diagnosis of 93.49% and for a negative diagnosis, 95.73%. The overall diagnostic accuracy was 94.84%, and the sensitivity of cytologic diagnosis was 98.75%. The false negative and false positive rates were 2.5%, including 10 cases with suspicious diagnoses and only 1 false positive diagnosis (0.23%). The false positive and false negative cases were recorded as having no effect or a minimal effect on patient management, and no mastectomy was performed because of this FNA diagnosis. FNA breast biopsy is an efficient tool for diagnosis when performed by surgical pathologists with cytologic training but not exclusively dedicated to cytopathology.


The effect of immunizing female rats with modified rat pituitary extract.
Immune reactions to a number of hormones have been induced, but information is lacking on the feasibility of inducing immune reactions to homologous gonadotropins. Female rats immunized with diazotized and tyrosylated rat pituitary extract containing gonadotropic activity and emulsified in complete Freund's adjuvant expressed various reproductive disturbances such as increased cycle length (7.0 days), increased number of sterile matings, reduced implantation in animals becoming pregnant, and reduced ability to carry fetuses to term. These effects were not exhibited by rats immunized similarly with diazotized or tyrosylated rat pituitary extract containing no detectable gonadotropic activity. Rats not becoming pregnant had a significantly higher antibody level to rat luteinizing hormone than did rats becoming pregnant. No significant elevation of rat follicle-stimulating hormone antibodies was noted. It is concluded that auto-immune reactions to gonadotropins can be induced and that very low levels of antibodies have a significant effect on reproduction.


Baseline asynchrony, assessed circumferentially using temporal uniformity of strain, besides coincidence between site of latest mechanical activation and presumed left ventricular lead position, predicts favourable prognosis after resynchronization therapy.
Traditional indexes of LV dyssynchrony (DYS) in pts to be resynchronized are sensitive to noise, while the concordance between LV lead position and site of latest mechanical activation is suggested to be, in these patients, clinically relevant. Both aspects, asynchrony and lead position have been addressed separately but unclear is their potential synergistic role in the clinical evolution of CRT patients. We assessed clinical and echocardiographic outcome, as well as mid-term prognosis, in a population of CHF patients submitted to CRT, stratified according to a novel asynchrony quantitation (temporal uniformity of strain: TUS) method and concordance or not between presumed LV lead position and site of latest mechanical activation. TUS was computed in 85 pts (QRS > 120 ms, EF < 0.35) in whom we measured circumferential and longitudinal strains using speckle-tracking 2D-echocardiography before and 3-6 months after CRT, together with triplane apical LV volumes. Optimal LV lead position in short axis view was defined as concordance of the segment with latest systolic circumferential strain prior-CRT and segment with assumed LV lead position. Assumed LV lead position was defined from a chest X-ray obtained 1 day after implantation and scored as anterior, lateral, posterior or inferior using 2 orthogonal views (antero-posterior and lateral). Following CRT, LV volume decreased (diastolic -8 ± 20%) and EF improved (+6 ± 9%, P < 0.001 for both). Two-way ANOVA revealed TUS improvement post-CRT (+22 ± 68%, P = 0.025), with a clear evidence for more marked asynchrony detectable at circumferential (from 0.53 ± 0.20 to 0.55 ± 0.19) as compared with longitudinal level (from 0.56 ± 0.14 to 0.62 ± 0.14) (P = 0.017). Multivariate analysis revealed that greater baseline asynchrony, as assessed circumferentially (P = 0.079), together with concordance between LV lead position and site of activation (P = 0.012), besides younger age (P = 0.051), longer QRS duration (P = 0.021) and higher baseline EF (P = 0.04),), but not longitudinal TUS (P = 0.231) did predict death from any cause or new episodes of pulmonary or systemic congestion requiring i.v. diuretics during a 529 ± 357 days clinical follow-up. We conclude that DYS indexed by circumferential TUS yields CRT benefits, supporting the idea of targeting TUS-measured DYS as the informative asynchrony quantitative measurement in CRT pts. Significant predictability in medium-term clinical follow-up of patients to be resynchronized is also associated with concordance between site of latest mechanical activation and presumed LV lead position in the present study.


Diagnostic RET genetic testing in 1,058 index patients: A UK centre perspective.
Diagnostic germline RET analysis is offered to all patients with a diagnosis of medullary thyroid carcinoma (MTC), or other conditions associated with multiple endocrine neoplasia type 2 (MEN2) in the United Kingdom. Here, we report the experience of a single centre's germline RET analysis over a 21-year period. Retrospective case-note review. All index patients referred to the Exeter Genomics Laboratory for diagnostic germline RET analysis between 1997 and 2018, and unaffected family members, undergoing predictive testing. The rate and nature of pathogenic variant detection were recorded, as well as the indication for testing. 1,058 index patients and 551 unaffected family members were tested. The overall rate of pathogenic variant detection was 10.2% amongst index patients and 29% amongst unaffected family members. The commonest indication was isolated MTC, and amongst the 690 patients with isolated MTC, 68 (9.9%) were found to harbour a RET pathogenic variant. Of those with presumed sporadic MTC, 8.5% were found to harbour germline RET pathogenic variants, compared with 36.4% of those with a family history of MEN2-associated conditions. Pathogenic variants were identified in 3.6% and 0% of patients with isolated phaeochromocytoma and primary hyperparathyroidism, respectively. Although the detection rate of RET germline pathogenic variants in patients with presumed sporadic MTC was significant, the overall detection rate in those with MTC was lower than expected in this series. Advances in RET analysis in response to reports of new variants over the last two decades are likely to have improved the pick-up rate in recent years.


The effect of dihydrotestosterone and culture conditions on proliferation of the human prostatic cancer cell line LNCaP.
Cell density, nutritional state, and serum factors modify the growth response of LNCaP human prostatic cancer cells to dihydrotestosterone. Evaluation of growth response to dihydrotestosterone requires logarithmic transformation of cell count or thymidine incorporation data. Under conditions of dose response, growth increases with cell density but no significant interaction of dihydrotestosterone with cell density was found under optimal culture conditions. The frequency of media change was a significant factor in modulating dose response. When cells from cultures maintained at different feeding periods were plated at different cell densities of (trypan blue) viable cells, significant effects of plating density on dihydrotestosterone response were found. Dihydrotestosterone protects cells under the adverse effects of media deprivation. Under the extreme adverse effects of serum deprivation, cells respond to dihydrotestosterone even under conditions of increasing cell loss. The effects of dihydrotestosterone on final cell density were significant. In the absence of serum, the elongated cells of LNCaP assume a round shape, but many remain adherent to the culture dish and can be restored to normal morphology by serum. A number of growth factors fail to restore normal morphology that was completely restored by a combination of fibronectin and dihydrotestosterone. We have not developed a practicable serum-free system for LNCaP.


The Association between Mothers' Smartphone Dependency and Preschoolers' Problem Behavior and Emotional Intelligence.
This study aimed to identify the level of mothers' smartphone dependency and determine its correlation with preschoolers' problem behavior and emotional intelligence. From 1 November to 30 December 2020, 141 mothers of preschool children (aged three to six years) were recruited to complete questionnaires that assessed their smartphone dependency and their child's problem behavior and emotional intelligence. The result revealed that the younger the mother and the higher the perception of boredom in daily living, the higher was the level of her smartphone dependency. Maternal smartphone dependency was also significantly correlated with the aggression, oppositional, and emotional instability subscales of the tool assessing children's problem behavior. To prevent problem behaviors among preschoolers, strategies to reduce mothers' smartphone dependency are needed.


Facial expression and imagery in depression: an electromyographic study.
When subjects are instructed to self-generate happy, sad, and angry imagery, discrete patterns of facial muscle activity can be detected using electromyographic (EMG) procedures. Prior research from this laboratory suggests that depressed subjects show attenuated facial EMG patterns during imagery conditions, particularly during happy imagery. In the present experiment, 12 depressed subjects and 12 matched normals were requested to generate happy and sad imagery, first with the instruction to simply "think" about the imagery, and then to self-regulate the affective state by "reexperiencing the feelings" associated with the imagery. Continuous recordings of facial EMG were obtained from the corrugator, zygomatic major, depressor anguli oris, and mentalis muscle regions. It was hypothesized that (a) these muscle sites would reliably differentiate between happy and sad imagery. (b) the instruction to self-generate the affective feeling state would produce greater EMG differences than the "think" instructions, and (c) the "think" instructions would be a more sensitive indicator of the difference between depressed and nondepressed subjects, especially for happy imagery. All three hypotheses were confirmed. The application of facial electromyography to the assessment of normal and clinical mood states, and the role of facial muscle patterning in the subjective experience of emotion, are discussed.


Prevalence and determinant factors of anemia in children aged 6-12 months after starting an iron supplement in the east of Iran.
Anemia in infants is a common problem, with regular iron supplementation the suggested policy for prevention and control of anemia. The aim of this study was to determine the prevalence and the determining factors of anemia as well as the benefits of regular iron supplementation in 6- to 12-month-old children. In this cross-sectional study, 897 children aged from 6 to 12 months cared for at the Tabas Health Centers, affiliated with Birjand University of Medical Sciences, South Khorasan, Iran, were enrolled in the study. Demographic and anthropometric data as well as the level of hemoglobin were collected through interviews and laboratory tests, respectively. Data analysis was performed using SPSS-22 and stata-13. Chi2, polychromic PCA, and logistic regression were used. The statistical significance level was 0.05. The prevalence of anemia, according to the WHO criterion for hemoglobin, was 36.8% (95% CI, 33.6-40.0). Its prevalence showed a decreasing trend as age and duration of iron supplement increased. Each 1-month increase in infant age was associated with a decreased risk of anemia [OR=0.88 (95% CI, 0.80-0.98)]. Anemia was not significantly different between genders. Individuals in the highest category for parity (≥3) were at a 2.3-fold greater risk of anemia compared with the lowest category [OR=2.35 (95% CI, 1.43-3.84)]. In contrast, individuals in the highest category for maternal age (>35 years) had a 62% lower risk of anemia compared with the reference category (<25 years) [OR=0.38 (95% CI, 0.20-0.72)]. Implementation of an iron supplementation plan in Iran has decreased anemia among 6- to 12-month-old children.


Quantitation of cellular components in Cryptococcus neoformans for system biology analysis.
Methods and procedures in molecular biology used to study fungal pathogenesis have significantly improved during the last decade. In this chapter, we provide step-by-step procedures for performing genetics and biochemical studies in the human pathogenic fungal microorganism Cryptococcus neoformans (Cn). These methods are employed for studying the pathobiology of Cn and for experimental validation of theoretical models of fungal pathogenicity.


Fibroblast growth factor 21 is an independent predictor of prevalent and incident obstructive sleep apnea.
Fibroblast growth factor 21 (FGF21) is a metabolic regulator and a potential biomarker of metabolic diseases. Limited data are available on the association between FGF21 and obstructive sleep apnea (OSA), which is considered as a manifestation of metabolic syndrome. In the present cross-sectional and longitudinal analyses, the FGF21 level was associated with OSA. This analysis of two clinical cohorts is the first to show that the FGF21 level increased significantly with OSA severity and was an independent predictor of incident OSA in Chinese adults. The circulating FGF21 level could serve as a potential serum biomarker of OSA and its comorbidities and thus aid risk evaluation and early intervention.


Motion sickness in amphibians.
We explored the question of whether amphibians get motion sickness by exposing anurans (frogs) and urodeles (salamanders) to the provocative stimulus of parabolic aircraft flight. Animals were fed before flight, and the presence of vomitus in their containers after flight was used to indicate motion-induced emesis. None of the species that we studied vomited during the 8 to 10 parabolas of each flight. However, at least one specimen from each of the anuran species Rana rugosa, Rana nigromaculata, Hyla japonica, and Rhacophorus schlegelii vomited in a period of 0.5 to 42 h after flight. Some specimens of R. nigromaculata, H. japonica, and R. schlegelii were also observed retching without emesis either during or shortly after exposure to parabolic flight. We were unable to induce either emesis or retching behavior in the aquatic from Xenopus laevis. Among the urodeles studied we saw no signs of motion sickness in either adult or larval Cynops pyrrhogaster, but at least one larval Hynobius nebulosus vomited shortly after parabolic flight. The amphibian species that exhibited the most motion sickness were the same ones that showed the greatest amount of tumbling during the microgravity phases of their parabolic flights. The most distinctive difference between motion sickness in amphibians and mammals that vomit, including man, is the long delay between a provocative stimulus and emesis proper in the amphibians. The retching behavior we induced in the frogs was identical to that described previously for frogs treated with emetic drugs. H. japonica, exposed to extended periods of microgravity on the MIR Space Station, flattened their bellies against the substrate and dorsiflexed their heads in a manner reminiscent of drug-induced nausea. In light of our current observations of retching behavior in motion sick H. japonica, we suggest that the previously observed behavior of three frogs on the MIR Space Station was a manifestation of motion sickness.


Establishment and characterization of two human cutaneous angiosarcoma cell lines, KU-CAS3 and KU-CAS5.
Cutaneous angiosarcoma (CAS) is a rare but fatal cancer. Established CAS cell lines are necessary for the investigation of their properties and treatment options. Two cell lines, KU-CAS3 and KU-CAS5, were established from human angiosarcoma specimens obtained from the scalp. Flow cytometric assay, tube formation assay, low-density lipoprotein (LDL) uptake assay, immunofluorescence analysis, real-time PCR, tumorigenesis assay, and STR analysis were conducted. The cells showed endothelial cell properties, based on the cobblestone appearance upon reaching confluence, CD31 positivity, tube-formation activity, active uptake of acetylated LDL, and vWF expression. The two cell lines expressed relatively high levels of adrenergic β2 receptor, and the VEGF1 and VEGF2 receptors. In the in vivo study, the growing neoplasms, confirmed as CAS, were identified as subcutaneous dark papules. KU-CAS cell lines were considered authentic based on STR profiling. KU-CAS3 and KU-CAS5 are the first human CAS cell lines having tumorigenic potential in vivo.


Evaluation of the effect of ambrisentan on digital microvascular flow in patients with systemic sclerosis using laser Doppler perfusion imaging: a 12-week randomized double-blind placebo controlled trial.
In patients with systemic sclerosis (SSc), digital ischemia results from an occlusive microvasculopathy that may not respond adequately to conventional vasodilators. Endothelin receptor antagonists can potentially modify the fibroproliferative vascular remodeling in SSc, and hence their use may be justified in the management of digital ischemia. The objective of this clinical trial was to evaluate the effect of ambrisentan, a selective endothelin type A receptor antagonist, on microvascular blood flow in patients with limited systemic sclerosis (SSc) using laser Doppler perfusion imaging (LDPI). In this randomized, double-blind, placebo controlled study we enrolled 20 patients with limited SSc. Fifteen patients received ambrisentan 5 mg daily for one month and then 10 mg daily for two months, and five received a placebo. There were three visits: weeks 0 (baseline), one and 12. Three patient-oriented questionnaires were completed at each visit: Scleroderma-Health Assessment Questionnaire (S-HAQ), Raynaud Condition Score (RCS), and Pain-Visual Analog Scale (P-VAS). At each visit, LDPI was used to obtain three blood flow readings involving regions of interest in second to fifth fingers of the non-dominant hand at room temperature (25°C) and after cooling (10°C) for two minutes. There were 16 females (80%); mean age was 50 years. None of the differences in blood flow (as measured by LDPI) were significant both at baseline and after cooling. However, patients in the ambrisentan group showed significant improvement in the patient-oriented outcomes: RCS (P=0.001) and S-HAQ score (P=0.005). This pilot study did not show evidence of significant increase in digital blood flow over time; however, there was an improvement in RCS and S-HAQ score. We conclude that continuous use of ambrisentan for three months does not seem to significantly improve digital blood flow in SSc patients. Clinicaltrials.gov NCT01072669. Registered 19 February 2010.


Exceeding body composition standards is associated with a more negative body image and increased weight cycling in active duty U.S. soldiers.
Negative body image is more often identified in individuals with overweight or obesity. More than 65% of U.S. soldiers have a body mass index (BMI) that classifies them with overweight or obesity. Additionally, all soldiers must meet body composition and physical fitness standards which may increase the risk of negative body image. This cross-sectional study aimed to identify associations between compliance with body composition standards, body image, and weight cycling through surveying 969 active-duty soldiers (86% male, 24.0 ± 5.5 years of age, BMI 26.0 ± 3.6 kg/m2). Compliance with body composition standards was measured by whether a soldier had ever taken and failed the circumference-based body fat assessment. Weight cycling was self-reported as ≥3 weight fluctuations of ≥5% of body weight during their military career. Multivariate linear and logistic regression models were used to examine the relationship between compliance with body composition standards, body image, weight cycling, and sociodemographic characteristics while controlling for BMI. Failing the circumference-based body fat assessment was associated with increased concern with conforming to military image. Negative body image was associated with higher odds of weight cycling. Further examination is warranted to understand the effects of body composition standards on soldiers' body image and weight cycling.


Absenteeism-Related Wage Loss Associated With Multimorbidity Among Employed Adults in the United States.
To estimate the incremental absenteeism-related wage loss associated with multimorbidity and examine the influence of paid sick leave policy (PSLP) on multimorbidity-associated absenteeism wage loss. We used the Medical Expenditure Panel Survey (MEPS) 2015 data. Two-part generalized linear models (GLM) were employed, using binomial distribution and gamma distribution with a log link. Nationally, multimorbidity was associated with a $9 billion incremental absenteeism-related wage loss annually among working adults. Absenteeism-related wage loss was higher among those with multimorbidity than those without multimorbidity. The incremental annual absenteeism-related wage loss associated with multimorbidity was lower in settings that offered paid sick leave than that did not offer paid sick leaves. Multimorbidity is associated with higher absenteeism-related wage loss. Paid sick leave policies can reduce the impact of multimorbidity on absenteeism-related wage loss.


Disposable laryngeal tube suction: standard insertion technique versus two modified insertion techniques for patients with a simulated difficult airway.
The disposable laryngeal tube suction (LTS-D) is a supraglottic airway device that can be used as an alternative to tracheal tube to provide ventilation. We tested the hypothesis that, with a frontal jaw thrust insertion technique (FIT/JT), the rate of correct placement attempts in patients with a simulated difficult airway by means of a rigid cervical immobilization collar could be significantly increased compared to the standard insertion technique (SIT) recommended by the manufacturer. 70 adult patients undergoing trauma surgery under general anaesthesia had an LTS-D inserted, randomly assigned to the SIT or FIT/JT. In the FIT/JT, the operator was standing in front of the patient's head, and forced chin lift to create sufficient retropharyngeal space was performed. The rate of successful tube placements within 180s and with a maximum of two attempts was the main outcome variable. To distinguish between the effects of the frontal approach and the jaw thrust manoeuvre, a third group was studied after completion of the SIT and FIT/JT groups. The standard insertion technique, but with a jaw thrust manoeuvre (SIT/JT), was employed in another 35 consecutive patients. Overall placement success was 49% (SIT, 17/35 patients, P<0.001), 91% (SIT/JT, 32/35 patients) and 100% (FIT/JT). The time required for successful insertion was shortest in the FIT/JT group (23±6s), and significantly longer in the SIT/JT (42±29s, P<0.001) and SIT groups (51±29s, P<0.0001). In anaesthetised patients with a simulated difficult airway created with a rigid cervical collar, the overall LTS-D placement success was significantly higher when a jaw thrust manoeuvre was performed, regardless of the particular technique used to introduce the LTS-D. Therefore, an intense jaw thrust manoeuvre should be performed whenever an LTS-D is being inserted.


Resolution of retinoschisis after vitreous surgery in X-linked retinoschisis.
To describe the efficacy of vitreous surgery to treat X-linked retinoschisis. Retrospective, comparative, interventional case series. Three patients (5 eyes) had foveal retinoschisis with peripheral retinoschisis. The patients' ages were 26, 7, and 17 years at the first surgery. The preoperative best-corrected visual acuity (BCVA) was 20/100 and 20/40 in patient 1, 20/30 in patient 2, and 20/100 and 20/200 in patient 3. The vitreous surgery consisted of core vitrectomy, surgically induced posterior vitreous detachment (PVD), removal of the internal limiting membrane (except for the right eye of patient 1), and 30% sulfur hexafluoride gas tamponade. The follow-up periods ranged from 6 months to 12 years. Best-corrected visual acuity and retinal tomography monitored by optical coherence tomography. Restoration of the foveal depression with collapse of the schisis cavity was achieved with the first surgery in 4 (80%) of the 5 eyes. In the right eye of patient 1, the retinoschisis persisted after the first surgery because of failed surgically induced PVD; the retinoschisis resolved promptly after the second surgery with successful induction of PVD. The BCVA improved in 3 eyes and was unchanged in 2 eyes. Vitreous surgery resulted in reattachment of the retinoschisis with visual improvement. Vitreous traction may play a role in the development of foveal retinoschisis.


Fludarabine modulates immune response and extends in vivo survival of adoptively transferred CD8 T cells in patients with metastatic melanoma.
Adoptive T cell therapy involving the use of ex vivo generated antigen-specific cytotoxic T lymphocytes provides a promising approach to immunotherapy. It has become increasingly apparent that anti-tumor efficacy using adoptively transferred T cells is linked to their duration of in vivo persistence and can only be achieved when combined with some form of pre-infusion patient conditioning regimen. An optimal conditioning regimen that provides a positive benefit without serious toxicities has yet to be defined. We have established a unique clinical model that allows for evaluation of a given conditioning regimen on adoptively transferred T cells in humans. In this first-in-human study (FHCRC #1796), we evaluate the use of fludarabine, an FDA-approved reagent with predictable lymphodepleting kinetics and duration of action, as a conditioning regimen that promotes homeostatic upregulation of cytokines and growth signals contributing to in vivo T cell persistence. We conducted a phase I study in patients with refractory metastatic melanoma. Patients received two infusions of a single tumor-reactive antigen-specific CTL clone expanded to 10(10)/m(2); the first infusion was given without fludarabine conditioning, and the second CTL infusion was given after a course of fludarabine (25 mg/m(2)/dayx5 days). This design permits intra-patient comparison of in vivo T cell persistence pre- and post-fludarabine. Nineteen CTL infusions were administered to ten patients. No serious toxicities were observed. Three of nine evaluable patients experienced minor response or stable disease for periods of 5.8-11.0 months with two additional patients demonstrating delayed disease stabilization. The median overall survival in this heavily pre-treated population was 9.7 months. Fludarabine led to a 2.9 fold improvement in the in vivo persistence of transferred CTL clones from a median of 4.5 days (range 0-38+) to 13.0 days (range 2-63+) (p<0.05). Fludarabine lymphodepletion increased plasma levels of the homeostatic cytokines IL-7 and IL-15. Surprisingly, fludarabine also increased the relative percentage of CD4+ T cells expressing the regulatory protein Foxp3. Lymphodepletion with fludarabine enhances transferred T cell persistence but suggest that additional improvements to optimize T cell survival and address regulatory T cells are critical in providing anti-tumor efficacy. ClinicalTrials.gov NCT00317759.