HIV/SIV DNA vaccine combined with protein in a co-immunization protocol elicits highest humoral responses to envelope in mice and macaques.
Vaccination with HIV/SIV DNAs elicits potent T-cell responses. To improve humoral immune responses, we combined DNA and protein in a co-immunization protocol using in vivo electroporation in mice and macaques. DNA&protein co-immunization induced higher antibody responses than DNA or protein alone, or DNA prime/protein boost in mice. DNA&protein co-immunization induced similar levels of cellular responses as those obtained by DNA only vaccination. The inclusion of SIV or HIV Env gp120 protein did not impair the development of cellular immune responses elicited by DNA present in the vaccine regimen. In macaques, the DNA&protein co-immunization regimen also elicited higher levels of humoral responses with broader cross-neutralizing activity. Despite the improved immunogenicity of DNA&protein co-immunization, the protein formulation with the EM-005 (GLA-SE) adjuvant further increased the anti-Env humoral responses. Dissecting the contribution of EM-005, we found that its administration upregulated the expression of co-stimulatory molecules and stimulated cytokine production, especially IL-6, by dendritic cells in vivo. These terminally differentiated, mature, dendritic cells possibly promote higher levels of humoral responses, supporting the inclusion of the EM-005 adjuvant with the vaccine. Thus, DNA&protein co-immunization is a promising strategy to improve the rapidity of development, magnitude and potency of the humoral immune responses.


Mechanisms of docetaxel resistance in prostate cancer: The key role played by miRNAs.
One of the main problems with the treatment of metastatic prostate cancer is that, despite an initial positive response, the majority of patients develop resistance and progress. In particular, the resistance to docetaxel, the gold standard therapy for metastatic prostate cancer since 2010, represents one of the main factors responsible for the failure of prostate cancer therapy. According to the present knowledge, different processes contribute to the appearance of docetaxel resistance and non-coding RNA seems to play a relevant role in them. In this review, a comprehensive overview of the miRNA network involved in docetaxel resistance is described, highlighting the pathway/s affected by their activity.


Flow injection immunoassay for carcinoembryonic antigen combined with time-resolved fluorometric detection.
Time-resolved fluorescence has been developed for immunoassay to obtain higher sensitivity than usual immunoassays. In this paper, a simple, sensitive and specific method was developed for immunoassay of serum carcinoembryonic antigen (CEA) by combining time-resolved fluoroimmunoassay (TRFIA) and flow injection analysis. Based on a sandwich immunoassay format, a monoclonal antibody immobilized immunoaffinity column inserted in a flow system was used for immunoreactions. The cleaved solution was collected after the reaction between the immunocomplex in the immunoaffinity column and the enhancement solution that was used to cleave the Eu-labels from the immunocomplex, and then detected by time-resolved fluorescence. Serum CEA could be detected in the linear range from 2.5 to 100 ng/ml with a correlation coefficient of 0.997 and the detection limit of 1.0 ng/ml. Twenty human serum samples detected by this method were in good agreement with the results obtained by the electrochemiluminescence immunoassay. This method could be further developed for fast practical clinical detection of serum CEA levels.


Fluorescent in-situ hybridization of cattle and sheep chromosomes with cloned human fragile-X DNA.
An extensive study on spontaneous and 5-Fluorodeoxyuridine induced fragile sites identified Xq31 in cattle (Bos taurus) and (Xq24, Xq26) in sheep (Ovis aries) in addition to several autosomal fragile sites (under publication). A ZOO-FISH study using three cloned human fragile-X probes with CCG/CGG(n) trinucleotide repeat sequence was carried out to determine homology between human and bovine fragile-X. The hybridisation results showed only a weak signal on a human chromosome that was not an X with all three fragile site probes. No signals were detected in sheep chromosomes. The signal of all three human fragile-X probes on cattle chromosomes was however, medium-prominent sub-centromeric signal on two homologues. BrdU administration in 12 h before harvesting identified these homologues to be chromosome number 5. In addition retrospective slides of cattle and sheep chromosomes used for fragile site studies showed no signals whatsoever. It was therefore concluded that no homology existed between human and bovine fragile-X.


Association between endothelial dysfunction and frailty: the Toledo Study for Healthy Aging.
Cardiovascular disease (CVD), both clinical and subclinical, has been proposed as one of the mechanisms underlying frailty. However, there is no evidence addressing the relationship between the earliest stage of CVD (endothelial dysfunction) and frailty. The goal of the study was to analyze the association between endothelial dysfunction, evaluated by asymmetric dimethylarginine (ADMA) levels, and frailty. We used data from the Toledo Study for Healthy Aging, a prospective Spanish cohort study. Biological samples were obtained and ADMA levels were determined using an enzyme immunoassay method. Logistic regression was used to estimate the odds ratio (OR) and 95 % confidence intervals of frailty associated with ADMA. Adjustments were made for age, gender, cardiovascular risk factors, and presence of atherosclerotic disease (assessed by ankle–brachial index; ABI). One thousand two hundred eighty-seven community-dwelling elderly were included. One hundred seven (8.3 %) were identified as frail, 542 (42.1 %) as pre-frail, and 638 (49.6 %) as non-frail. ADMAvalues were higher in frail subjects than in non-frail ones. In addition, an interaction between the presence of atherosclerotic disease and ADMA on the odds of frailty (p=0.045) was detected. After adjustments for age, classical cardiovascular risk factors, and ABI, the risk of frailty was associated with increasing levels of ADMA in subjects without atherosclerotic disease [OR for 1 standard deviation increase in ADMA=1.14 (1.01–1.28), p=0.032] but not in those with atherosclerotic disease. In our study, endothelial dysfunction, assessed by ADMA levels, is associated with frailty. These findings provide additional support for a relevant role of vascular system since its earliest stage in frailty.


Detection of a neurotoxic quaternary pyridinium metabolite in the liver of haloperidol-treated rats.
Various theories have been proposed in the past to explain the side effects associated with haloperidol treatment. In this study, we tested the hypothesis that in vivo biotransformation of haloperidol to a quaternary pyridinium metabolite might contribute to some of these effects. Administration of haloperidol (IP or by gavage) to male Wistar rats results in its biotransformation to a butyrophenone quaternary pyridinium metabolite similar to MPTP metabolism to MPP+. The corresponding methyl quaternary pyridinium compound was not detected in the liver or brain of the rats used in this study. The proposed methyl quaternary pyridinium compound and the observed butyrophenone metabolite were tested for neurotoxicity in a frog model of MPTP action and were found to have neurotoxic effects, although the methyl quaternary pyridinium compound was much more toxic.


Assessment of paraoxonase 1 activity and malondialdehyde levels in patients with rheumatoid arthritis.
We aimed to evaluate antioxidant paraoxonase 1 activity together with malondialdehyde (MDA) (an oxidative stress parameter) levels in patients with rheumatoid arthritis. Fifty-seven rheumatoid arthritis patients were included in the study and subgrouped according to disease activity (active, n = 31; inactive, n = 26) and compared with healthy controls (n = 25). Serum paraoxonase 1 activity and MDA levels were measured according to an enzymatic spectrophotometric method. Serum MDA level was higher (P = 0.001) whereas paraoxonase 1 activity was lower (P = 0.001) in the patient group than the controls. When active and inactive subgroups were compared with the control group, there was a statistically significant difference between each parameter. Serum MDA levels were significantly higher, while paraoxonase 1 activity was lower in the active and inactive rheumatoid arthritis groups than the control group. But there was not any difference between active and inactive patients with RA. There was a negative correlation between MDA levels and paraoxonase 1 activity. Increased reactive oxygen species levels in rheumatoid arthritis may result in a pro-oxidation environment, which in turn could result in decreased antioxidant paraoxonase 1 activity and increased MDA levels.


Adsorption and desorption performance of benzene over hierarchically structured carbon-silica aerogel composites.
Hierarchically structured carbon-silica aerogel (CSA) composites were synthesized from cheap water glass precursors and granulated activated carbon via a post-synthesis surface modification with trimethylchlorosilane (TMCS) and a low-cost ambient pressure drying procedure. The resultant CSA composites possess micro/mesoporous structure and hydrophobic surface. The adsorption and desorption performance of benzene on carbon-silica aerogel composite (CSA-2) under static and dynamic conditions were investigated, comparing with pure silica aerogel (CSA-0) and microporous activated carbon (AC). It was found that CSA-2 has high affinity towards aromatic molecules and fast adsorption kinetics. Excellent performance of dynamic adsorption and desorption observed on CSA-2 is related to its higher adsorption capacity than CSA-0 and less mass transfer resistance than AC, arising from the well-developed microporosity and open foam mesostructure in the CSA composites.


Increasing incidence of childhood thyrotoxicosis in a population-based area of central Sweden.
To study the incidence, prevalence and clinical characteristics of childhood thyrotoxicosis in a population-based area comprising five counties in central Sweden. Retrospective analysis of the medical records of all children below 16 y of age with thyrotoxicosis during a 10-y study period. Forty-six children were identified with thyrotoxicosis. The median age at onset was 11.7 y and 85% were females. At the end of the study period on 31 December 1999, the prevalence was 0.08 per 1000. The mean annual incidence was four times higher in the second 5-y period than in the first (2.7 and 0.7 per 100000, respectively; p < 0.001). A first-degree relative with thyrotoxicosis was found in 21% of the children and 11/46 children (24%) showed autoimmune-related co-morbidity. Increased heart rate was observed in 95%, goitre in 67% and exophthalmos in 33% of the children. TSH was completely suppressed and free T4 was clearly above the upper reference limit in all children. TSH receptor-stimulating antibodies and peroxidase antibodies were observed in 80% and 71% of the children, respectively. This study indicates an increase in the incidence of childhood thyrotoxicosis in a population-based area of central Sweden. A prospective study aimed at identifying specific aetiological risk factors for development of thyrotoxicosis has been initiated.


Norepinephrine content of discrete brain nuclei in acutely and chronically stressed borderline hypertensive rats.
Forty-three male borderline hypertensive rats were subjected to either 3 days, or 4, 10, or 16 weeks of daily stress. An additional 43 animals served as unstressed, age-matched controls. At the end of study, animals were sacrificed, brains were removed, and cardiovascularly-important nuclei in the brainstem and hypothalamus were removed by micropunch. Assays revealed that norepinephrine (NE) levels were initially elevated in the brainstem in animals stressed for 3 days. As stress continued, NE levels were significantly lower in the brainstem, and eventually in the hypothalamus, of stressed animals. The relationship of these observations to environmentally-induced hypertension is discussed.


Theophylline plasma clearance in critically ill geriatric patients receiving total parenteral nutrition and in control patients.
Mean values and standard deviations are reported for theophylline plasma clearance in 45 hospitalized geriatric patients (age greater than 65 years), distributed into two groups: Group I (n = 24) included critically ill patients admitted to the ICU on total parenteral nutrition (TPN). Group II (n = 21) included patients admitted to the Internal Medicine Ward without TPN. All patients were treated with a constant intravenous infusion of aminophylline. Group I patients had a lower mean theophylline clearance of 0.42 +/- 0.23 ml/kg/min compared to Group II 0.64 +/- 0.24 ml/kg/min (P = 0.003).


Quality of life and sexual problems in disease-free survivors of cervical cancer compared with the general population.
The purpose of this study was to identify problems related to long-term quality of life (QOL) and sexual function in cervical cancer survivors. The authors enrolled 860 women (median time since diagnosis, 5.86 years) with a history of cervical cancer (stage I to IVa) who had been treated at any of 6 hospitals from 1983 through 2004 and 494 control subjects selected randomly from a representative sample of Korean women. Subjects filled out a questionnaire that included the European Organization for Research and Treatment of Cancer (EORTC) QLQ-C30, its Cervical Cancer Module, and additional sexual function items. Cervical cancer survivors had clinically significant worse problems with social functioning, constipation, diarrhea, and difficulties with their finances than controls (P < .01). Survivors also reported more severe lymphedema and menopausal symptoms and worse body image, sexual and/or vaginal functioning, and sexual worry (P < .01). Anxiety about sexual performance was more problematic in survivors than in controls (P < .01), as was dyspareunia for women who received radiotherapy (P < .01). These findings can increase the awareness of healthcare providers to the potential need for counseling and other interventions among women who have been successfully treated for cervical cancer and could help them improve their impaired QOL.


Practice variation in bilateral salpingo-oophorectomy at benign abdominal hysterectomy: a population-based study.
Bilateral salpingo-oophorectomy at benign hysterectomy is not recommended in premenopausal women who are in the premenopausal stage because of its potential associations with increased all-cause mortality and cardiovascular disease; however, contemporary practice patterns are unknown. This study aimed to quantify between-surgeon variation in bilateral salpingo-oophorectomy and identify surgeon and patient characteristics associated with bilateral salpingo-oophorectomy to evaluate current quality of care and identify targets for knowledge translation and future research. We conducted a population-based retrospective cross-sectional study of adult women (≥20 years) undergoing benign abdominal hysterectomy from 2014 to 2018 in Ontario, Canada. Hierarchical multivariable logistic regression models, stratified by age group (<45, 45-54, ≥55 years), were used to model between-surgeon variation after multivariable adjustment for patient and surgeon characteristics. Cases of bilateral salpingo-oophorectomy were classified as potentially appropriate or potentially avoidable based on the presence or absence of diagnostic indications. Of 44,549 eligible women, 17,797 (39.9%) underwent concurrent bilateral salpingo-oophorectomy, and 26,752 (60.1%) did not. In all three age strata, the individual surgeon providing care was one of the strongest factors influencing whether patients received bilateral salpingo-oophorectomy (median odds ratio, 2.00-2.53). Surgeons accounted for more than 22% of the residual observed variation in bilateral salpingo-oophorectomy in women aged 45-54 years compared with 16% and 14% of the residual observed variation in bilateral salpingo-oophorectomy in women aged <45 and ≥55 years, respectively. Non-gynecologic patient factors, such as obesity (odds ratio, 1.33; 95% confidence interval, 1.17-1.52; P<.001) and residing in low-income regions (odds ratio, 1.34; 95% confidence interval, 1.16-1.55; P<.001), were also associated with bilateral salpingo-oophorectomy. Approximately 40% of patients who underwent bilateral salpingo-oophorectomy had no indication for the procedure in their discharge records. Marked between-surgeon variation in bilateral salpingo-oophorectomy rates, even after adjusting for patient case mix, suggests ongoing uncertainty in practice. Stronger evidence-based guidelines on the risks and benefits of salpingo-oophorectomy as women age are needed, particularly focusing on perimenopausal women.


Receptor preferences of cysteinyl-leukotrienes in the guinea pig lung parenchyma.
Two cysteinyl-leukotriene receptors, CysLT(1) and CysLT(2) receptors, have been cloned, but the contractions to cysteinyl-leukotrienes in the guinea pig lung parenchyma have been reported to be resistant to CysLT(2) receptor antagonism and to be only partially inhibited by CysLT(1) receptor antagonism. The receptor preferences of the individual cysteinyl-leukotrienes (leukotriene C(4), D(4) and E(4)) in the guinea pig lung parenchyma were studied in organ baths. CysLT(1) receptor antagonists competitively inhibited the contraction to leukotriene E(4), but exhibited only weak antagonism of contractions to leukotriene C(4) and D(4). In the presence of the cyclooxygenese inhibitor indomethacin and the nitric oxide synthase inhibitor N(omega)-nitro-L-arginine (L-NOARG), the CysLT(1) receptor antagonists did not further inhibit the leukotriene D(4)-induced contraction. These results suggest that leukotriene E(4) solely activates a CysLT(1) receptor, and that the CysLT(1) receptor antagonist-resistant contraction to leukotriene D(4) and C(4) is mediated via another CysLT receptor.


Monitoring of aerosols and studying its effects on the environment and humans health in Iran.
Natural hazards affect different parts of living organisms. One of these hazards is aerosols. Addressing this issue in areas that suffer from this hazard is of critical importance. Scientific research over the past two decades has shown that aerosol particles are one of the main pollutants from the perspective of public health and health. The purpose of the present study is to monitor and model aerosol temporal and spatial variations in Iran. Aerosols or airborne particles with health effects such as heart, vascular and respiratory diseases are associated. For this purpose, MODIS (or Moderate Resolution Imaging Spectroradiometer) and NOAA (or National Oceanic and Atmospheric Administration) satellite data and BTD (or Brightness Temperature Difference) were used. Given the 20-year study period (2000-2019), the output of much satellite data was divided into four five-year periods to monitor aerosols with high accuracy. The results showed that aerosol values in terms of intensity and frequency of optical depth (AOD) increased over the 20-year time series, and its intensity was higher in the last 5 years. According to the results obtained from the NODIS and NOAA satellite data and comparing their outputs, NOAA satellite data were associated with outliers, which was significantly different from the other cohort data. For this reason, the MODIS satellite image output was used to monitor aerosol images. The innovation of the present study is the use of remote sensing science to monitor the effects of aerosols on the environment and human's health. According to the results from MODIS satellite data, the maximum optical depth (AOD) of the aerosols is for July 2003 with a value of 0.63, but according to the NOAA satellite data output, the maximum optical depth (AOD) of the aerosols is for March 2013 with a value of 2.54. As the values of aerosols increase in frequency and intensity and the areas with the highest intensity of aerosols have been identified, it can be overcome by careful planning of their problems. Areas, where the amount and volume of aerosols were higher, should be observed in health protocols. By preventing and maintaining good hygiene, the negative effects of aerosols can be reduced.


Domain-specific associations between disability and depression, anxiety, and somatization in primary care patients.
This study explores the associations between different disability domains and the most prevalent symptoms of mental disorders in primary care patients (i.e. depression, anxiety, and somatization). A total of 1241 participants from 28 primary care centres completed self-report measures of depression, anxiety, and somatization. This same sample also completed the Sheehan Disability Scale (SDS) to assess functional impairment in work, social life, and family life domains. Associations between the symptoms and each disability domain were examined using hierarchical regression analyses. Depression emerged as the strongest predictor of all three disability domains. Somatization was associated only with the work domain, and anxiety was associated only with the family life domain. Clinical symptoms explained a greater proportion of the variance than sociodemographic variables. In primary care patients, depression, anxiety and somatizations were associated with distinct domains of disability. Early provision of effective treatments in the primary care setting may be crucial to reduce the societal burden of common mental disorders.


Dual role of arginine metabolism in establishing pathogenesis.
Arginine is an integral part of host defense when invading pathogens are encountered. The arginine metabolite nitric oxide (NO) confers antimicrobial properties, whereas the metabolite ornithine is utilized for polyamine synthesis. Polyamines are crucial to tissue repair and anti-inflammatory responses. iNOS/arginase balance can determine Th1/Th2 response. Furthermore, the host arginine pool and its metabolites are utilized as energy sources by various pathogens. Apart from its role as an immune modulator, recent studies have also highlighted the therapeutic effects of arginine. This article sheds light upon the roles of arginine metabolism during pathological conditions and its therapeutic potential.


Personalized persuasion: Quantifying susceptibility to information exploitation in spear-phishing attacks.
Many cyberattacks begin with a malicious email message, known as spear phishing, targeted at unsuspecting victims. Although security technologies have improved significantly in recent years, spear phishing continues to be successful due to the bespoke nature of such attacks. Crafting such emails requires attackers to conduct careful research about their victims and collect personal information about them and their acquaintances. Despite the widespread nature of spear-phishing attacks, little is understood about the human factors behind them. This is particularly the case when considering the role of attack personalization on end-user vulnerability. To study spear-phishing attacks in the laboratory, we developed a simulation environment called SpearSim that simulates the tasks involved in the generation and reception of spear-phishing messages. Using SpearSim, we conducted a laboratory experiment with human subjects to study the effect of information availability and information exploitation end-user vulnerability. The results of the experiment show that end-users in the high information-availability condition were 2.97 times more vulnerable to spear-phishing attacks than those in the low information-availability condition. We found that access to more personal information about targets can result in attacks involving contextually meaningful impersonation and narratives. We discuss the implications of this research for the design of anti-phishing training solutions.


Early prophylactic anticoagulation via transjugular intrahepatic route for portal vein thrombosis after splenectomy in cirrhotic portal hypertension.
To evaluate early transcatheter anticoagulation via the transjugular intrahepatic route to prevent portal vein thrombosis (PVT) after splenectomy in cirrhotic patients with portal hypertension. This retrospective study included 98 cirrhotic patients with portal hypertension who underwent open splenectomy (48 men and 50 women; age, 45.4 y ± 13.6). Systemic anticoagulation was given to 52 patients in group I, and transcatheter anticoagulation was performed in 46 patients in group II. The technical success rate of catheterization by the transjugular intrahepatic route was 93.5% in group II. The 30-day (6.52% vs 23.1%, P < .05) and 6-month (8.70% vs 26.9%, P < .05) incidences of PVT were significantly lower in group II than in group I. The postoperative bleeding rate was 6.52% in group II and 25% in group I (P < .05). There was no significant difference between groups in 30-day (5.77% vs 2.17%) and 6-month (1.92% vs 6.52%) mortality. After splenectomy, the portal trunk vessel diameter was 16.0 mm ± 3.5 in group I and 14.5 mm ± 2.5 in group II (P < .05). The portal flow velocity was 25.9 cm/s ± 7.1 in group I and 28.2 cm/s ± 5.3 in group II (P > .05). During the first week after splenectomy, notable hypercoagulability was detected within the portal vein compared with peripheral blood. Decreased portal flow velocity was considered an independent risk factor for PVT by univariate and multivariate analysis. Transcatheter anticoagulation via the transjugular intrahepatic route can decrease the incidence of PVT and postoperative bleeding after open splenectomy in cirrhotic patients with portal hypertension.


Genomic Profiling of Childhood Tumor Patient-Derived Xenograft Models to Enable Rational Clinical Trial Design.
Accelerating cures for children with cancer remains an immediate challenge as a result of extensive oncogenic heterogeneity between and within histologies, distinct molecular mechanisms evolving between diagnosis and relapsed disease, and limited therapeutic options. To systematically prioritize and rationally test novel agents in preclinical murine models, researchers within the Pediatric Preclinical Testing Consortium are continuously developing patient-derived xenografts (PDXs)-many of which are refractory to current standard-of-care treatments-from high-risk childhood cancers. Here, we genomically characterize 261 PDX models from 37 unique pediatric cancers; demonstrate faithful recapitulation of histologies and subtypes; and refine our understanding of relapsed disease. In addition, we use expression signatures to classify tumors for TP53 and NF1 pathway inactivation. We anticipate that these data will serve as a resource for pediatric oncology drug development and will guide rational clinical trial design for children with cancer.


Combination therapy versus monotherapy for pulmonary arterial hypertension: a meta-analysis.
Several randomised controlled studies and a previous meta-analysis have reported conflicting results regarding the effect of combined targeted therapy compared with monotherapy for pulmonary arterial hypertension (PAH). We did a systematic review and meta-analysis to assess the effects of a combination of PAH-specific therapies compared with monotherapy on predefined clinical worsening in PAH. We searched MEDLINE, Embase, and the Cochrane Library for reports published from Jan 1, 1990, to May 31, 2015, of prospective randomised controlled trials of at least 12 weeks that assessed a combination of PAH-specific therapies (upfront and sequential add-on) compared with background PAH-specific monotherapy in patients older than 12 years. We extracted data from the reports, and assessed the primary outcome of risk of clinical worsening, as defined a priori in each trial, using the Mantel-Haenszel method based on a fixed-effects model. Of 2017 studies that we identified from our search, we included 17 (4095 patients) in our analysis. 15 studies assessed clinical worsening and were included in the primary analysis. Combined therapy was associated with significant risk reduction for clinical worsening compared with monotherapy (combined therapy 17% [332 of 1940 patients] vs monotherapy 28% [517 of 1862 patients], risk ratio [RR] 0·65 [95% CI 0·58-0·72], p<0·00001). We noted no heterogeneity between the studies (I(2)=18%, phomogeneity=0·25). A publication bias was suggested by the results of an Egger test (t=-2·3982, p=0·031), but when we excluded the four studies with the highest SEs, the RR for clinical worsening was identical (0·65 [95% CI 0·58-0·73], p<0·00001). In our analysis, combined therapy for PAH was associated with a significant reduction in clinical worsening compared with monotherapy. However, our study was limited by the variable definition of clinical worsening among the trials and possible publication bias. Because many patients still had clinical worsening with combination therapy, identification of innovative therapeutic targets for PAH is thus urgently needed. None.


In vivo transformation of factor-dependent hemopoietic cells: role of intracisternal A-particle transposition for growth factor gene activation.
Cells of the granulocyte-macrophage colony stimulating factor (GM-CSF) or multi-lineage colony stimulating factor (Multi-CSF) dependent line FDC-P1 undergo leukemic transformation after injection into irradiated DBA/2 mice. About one third of factor-independent FDC-P1 variants isolated from leukemic animals express GM-CSF or Multi-CSF, assessed either by bioassay or by sensitive RNA detection using the polymerase chain reaction. All of the GM-CSF-secreting lines studied had a rearrangement in one allele of the GM-CSF gene, three of four Multi-CSF-secreting lines had Multi-CSF gene rearrangements, while factor-independent lines lacking evidence of growth factor production had no demonstrable CSF gene alterations. All rearrangements were characterized by insertions of novel DNA in the 5'-flanking regions of the CSF genes. The inserted segments of DNA varied in size between 0.35 and 6.5 kb and displayed restriction enzyme cleavage maps reminiscent of intracisternal A-particle (IAP) genomes. This was confirmed in two cases by molecular cloning and nucleotide sequence analysis. In these instances, the insertion consisted of solitary IAP long terminal repeats. The transformation system described provides a model for the study of IAP transpositions and their effects on gene activation.


Labor Analgesia: A Systematic Review and Meta-Analysis of Non-Pharmacological Complementary and Alternative Approaches to Pain during First Stage of Labor.
the aim of the study was to conduct a meta-analysis to evaluate the efficacy of non-invasive and non-pharmacological techniques on labor first-stage pain intensity. Literature databases were searched from inception to May 2021, and research was expanded through the screening of previous systematic reviews. Inclusion criteria were: (1) population: women in first stage of labor; (2) intervention: non-pharmacological, non-invasive, or minimally invasive intrapartum analgesic techniques alternative and/or complementary to pharmacological analgesia; (3) comparison: routine intrapartum care or placebos; (4) outcomes: subjective pain intensity; and (5) study design: randomized controlled trial. Risk of bias of included studies was investigated, data analysis was performed using R version 3.5.1. Effect size was calculated as difference between the control and experimental groups at posttreatment in terms of mean pain score. A total of 63 studies were included, for a total of 6146 patients (3468 in the experimental groups and 2678 in the control groups). Techniques included were massage (n = 11), birth balls (n = 5) mind-body interventions (n = 8), heat application (n = 12), music therapy (n = 9), dance therapy (n = 2), acupressure (n = 16), and transcutaneous electrical nerve stimulation (TENS) (n = 8). The present review found significant evidence in support of the use of complementary and alternative medicine for labor analgesia, and different methods showed different impact. However, more high-quality trials are needed.


Both chloride-binding sites are required for KCC2-mediated transport.
The K+-Cl- cotransporter 2 (KCC2) plays an important role in inhibitory neurotransmission, and its impairment is associated with neurological and psychiatric disorders, including epilepsy, schizophrenia, and autism. Although KCCs transport K+ and Cl- in a 1:1 stoichiometry, two Cl- coordination sites were indicated via cryo-EM. In a comprehensive analysis, we analyzed the consequences of point mutations of residues coordinating Cl- in Cl1 and Cl2. Individual mutations of residues in Cl1 and Cl2 reduce or abolish KCC2WT function, indicating a crucial role of both Cl- coordination sites for KCC2 function. Structural changes in the extracellular loop 2 by inserting a 3xHA tag switches the K+ coordination site to another position. To investigate, whether the extension of the extracellular loop 2 with the 3xHA tag also affects the coordination of the two Cl- coordination sites, we carried out the analogous experiments for both Cl- coordinating sites in the KCC2HA construct. These analyses showed that most of the individual mutation of residues in Cl1 and Cl2 in the KCC2HA construct reduces or abolishes KCC2 function, indicating that the coordination of Cl- remains at the same position. However, the coupling of K+ and Cl- in Cl1 is still apparent in the KCC2HA construct, indicating a mutual dependence of both ions. In addition, the coordination residue Tyr569 in Cl2 shifted in KCC2HA. Thus, conformational changes in the extracellular domain affect K+ and Cl--binding sites. However, the effect on the Cl--binding sites is subtler.


The higher toxicity of cereulide relative to valinomycin is due to its higher affinity for potassium at physiological plasma concentration.
Valinomycin and cereulide are bacterial toxins with closely similar chemical structure and properties but different toxic effects. Emetic poisoning is induced by cereulide but not by valinomycin. Both are specific potassium ionophores. Such compounds may affect mitochondrial functions. Both compounds cause a potassium-dependent drop in the transmembrane inner membrane potential due to the uptake of K+ as positively charged ionophore complex. Valinomycin is more potent than cereulide at high [K+] (>80 mM), whereas cereulide in contrast to valinomycin is active already at <1 mM. With cereulide, there is a substantial lag, while valinomycin acts without lag. Both ionophores induce mitochondrial swelling in the presence of K+, in the case of cereulide with a lag. These toxins strongly inhibited respiration at the level of complex IV when used at higher concentrations than that used for detection of ionophoretic transport of K+. At high [KCl] (120 mM), valinomycin was more potent than cereulide both as ionophore and inhibitor, but at low [KCl] (2.5 mM), cereulide was much more potent. Thus, valinomycin needed 20-30 mM KCl for substantial effects, cereulide only 1-3 mM K+, which is close to its level in blood serum. This explains the higher toxicity of cereulide at low concentrations with the positively charged potassium complex being accumulated in the cell by transport through the plasma membrane driven by the membrane potential. Furthermore, with similar concentrations, the final concentration of cereulide in the cells may become higher than that of valinomycin.


The effect of methallibure (I.C.I. 33,828) on the steriodogenesis in the ovary and testis of a fresh water teleost, Cyprinus carpio.
Methallibure treatment of mature Cyprinus carpio causes a marked reduction in the steroidogenesis, as indicated by the fall in 3beta-HSD activity in the gonads. Significance of these results is discussed.


Identification of apamin binding sites in rat intestinal mucosa.
Apamin, a specific blocker of one class of Ca(2+)-activated K+ channes, was used to detect the apamin receptors associated with K+ channels in the mucosa of the rat jejunum and colon. Two receptor sites for 125I-apamin have been identified. These sites differed in their affinity for apamin (jejunum: KD1 = 1.1 nM and KD2 = 170 nM; colon: KD1 = 0.5 nM and KD2 = 1.1 nM and KD2 = 140 nM) and the maximum number of sites (jejunum: B(max1) = 111 and B(max2) = 4030; colon: B(max1) = 187 and B(max2) = 7550 fmol/mg of protein). 125I-apamin binding was stimulated by K+ ions with K0.5 = 1.0 mM and inhibited by the neuromuscular blocker tubocurarine (KI = 50 microM). We interpret these data to demonstrate that the high-affinity, low-capacity binding sites reflect the existence of apamin-sensitive K+ channels in the intestinal mucosa.


An evaluation of training in family health for medical students and internes in Sri Lanka.
The results of an evaluation of an integrated training programme in family health for medical students and intern medical officers in Sri Lanka are presented. The course objectives were translated into expected student competences in specific tasks and the responses of the internes obtained using a semi-structured questionnaire. While the course had reinforced the students' awareness and commitment on the need for actively promoting family health it had not provided the expected competences in many tasks to the level of confidence that was anticipated. A number of deficiencies in the training programme in medical school as well as during the medical internship become evident and these have been discussed with the respective authorities with a view to effecting satisfactory improvements.


Prognostic value of bcl-2 immunoreactivity in medullary thyroid carcinoma.
Bcl-2 expression has been evaluated immunocytochemically in a series of 33 medullary thyroid carcinomas (MTC) with long-term (mean, 10.3 years) follow-up. Twenty-six of 33 cases showed intense bcl-2 immunoreactivity in more than 25% neoplastic cells. Bcl-2 immunoreactivity did not correlate with several clinicopathologic parameters including sex and age of the patients, sporadic or familial disease, tumor size and stage, amount of amyloid stroma, and immunoreactivity for calcitonin, chromogranin A, proliferating cell nuclear antigen (PCNA), N-myc, and p53. Lack of bcl-2 immunoreactivity, however, correlated significantly (P = .0001) with a shorter survival. Indeed, the seven patients with tumors devoid of bcl-2 immunoreactivity all died of disease within 8 years from the diagnosis. In multivariate analysis, lack of bcl-2 immunoreactivity was an independent predictor of worse prognosis (P = .001 for disease-free survival and P = .0001 for overall survival). None of the other clinicopathologic variable investigated proved to be an independent prognostic parameter. It is concluded that down-regulation of bcl-2 expression in MTC may identify a subset of tumors with a more aggressive clinical course.


Comparative studies of chromosomal aberration and mutagenicity of the trivalent and hexavalent chromium.
The comparative cytogenetic and mutagenic effects between trivalent and hexavalent chromium were investigated. Five chromium compounds, K2Cr2O7 and K2CrO4 containing Cr6+, and Cr(CH3COO)3, Cr(NO3)3 and CrCl3 containing Cr3+, were examined for their ability to induce chromosomal damage in cultures of human leukocytes, for their reactivity with DNA by a rec-assay system and for mutagenicity in the E. coli Hs30R test system. Chromosome-breaking activity was significantly higher for the compounds with hexavalent than trivalent chromium, the efficiency being in the decreasing order K2Cr2OM greater than K2CrO4 greater than Cr(CHCOO)3 greater than Cr(NO3)3, CrCl3. In the rec-assay and mutation assay, hexavalent (K2Cr2O7 and K2CrO4) and trivalent Cr(CH3COO)3) compounds gave positive results, their mutagenic potential being higher in the same order of clastogenic magnitude.


Structural analysis of the activation-induced deoxycytidine deaminase required in immunoglobulin diversification.
Activation-induced deoxycytidine deaminase (AID) initiates somatic hypermutation (SHM) and class-switch recombination (CSR) by deaminating C→U during transcription of Ig-variable (V) and Ig-switch (S) region DNA, which is essential to produce high-affinity antibodies. Here we report the crystal structure of a soluble human AID variant at 2.8Å resolution that favors targeting WRC motifs (W=A/T, R=A/G) in vitro, and executes Ig V SHM in Ramos B-cells. A specificity loop extending away from the active site to accommodate two purine bases next to C, differs significantly in sequence, length, and conformation from APOBEC proteins Apo3A and Apo3G, which strongly favor pyrimidines at -1 and -2 positions. Individual amino acid contributions to specificity and processivity were measured in relation to a proposed ssDNA binding cleft. This study provides a structural basis for residue contributions to DNA scanning properties unique to AID, and for disease mutations in human HIGM-2 syndrome.


A New Durable Surface Nanoparticles-Modified Perovskite Cathode for Protonic Ceramic Fuel Cells from Selective Cation Exsolution under Oxidizing Atmosphere.
A high-performance cathode of a protonic ceramic fuel cell (PCFC) should possess excellent oxygen reduction reactivity, high proton/oxygen-ion/electron conductivity, and sufficient operational stability, thus requiring a delicate tuning of both the bulk and surface properties of the electrode material. Although surface modification of perovskites with nanoparticles from reducing-atmosphere exsolution has been demonstrated effective at improving the electrochemical anodic oxidation, such nanoparticles would easily re-incorporate into the perovskite lattice causing a big challenge for their application as a cathode. Here, a durable perovskite-based nanocomposite cathode for PCFCs is reported, which is facilely prepared via the exsolution of nanoparticles in an oxidizing atmosphere. Through composition and cation nonstoichiometry manipulation, a precursor with the nominal composition of Ba0.95 (Co0.4 Fe0.4 Zr0.1 Y0.1 )0.95 Ni0.05 O3-δ (BCFZYN-095) is designed, synthesized, and investigated, which, upon calcination, gives rise to the formation of a perovskite-based nanocomposite comprising a major perovskite phase and a minor NiO phase enriched on the perovskite surface. The major perovskite phase enabled by the proper cation nonstoichiometry manipulation promotes bulk proton conduction while the NiO nanoparticles facilitate the oxygen surface exchange process, leading to a superior cathodic performance with a maximum peak power density of 1040 mW cm-2 at 650 °C and excellent operational stability of 400 h at 550 °C.


Home and Community-Based Service Use by Vulnerable Older Adults.
The purpose of this study was to identify different types of clients who use home and community-based services. Enrollment characteristics of 76 clients at risk of nursing home placement and Medicaid spend-down who were enrolled in the Virginia Community Living Program were analyzed. Two-step cluster analysis identified 4 groups of service users. Enabling resources (caregiver relationship to participant, participant living arrangement, and length of time caregiver provided assistance to participant) and disability type (physical, cognitive, traumatic brain injury, or other) differentiated the client groups. Groups differed on average service cost per day and likelihood of nursing home placement if services were not provided. Findings point to the value of having practitioners assist vulnerable clients in tailoring services to meet different care needs and the need for refining policies guiding home and community-based care.


A vibrational spectroscopic study of planchéite Cu₈Si₈O₂₂(OH)₄·H₂O.
Planchéite Cu(8)Si(8)O(22)(OH)(4)·H(2)O is a hydrated copper hydroxy silicate. Raman spectroscopy complimented with infrared spectroscopy has been used to determine the molecular structure of planchéite. Raman bands at around 1048, 1081 and 1127 are described as the ν(1)-SiO(3) symmetric stretching vibrations; Raman bands at 828, 906 are attributed to the ν(3)-SiO(3) antisymmetric stretching vibrations. The Raman band at 699 cm(-1) is assigned to the ν(4) bending modes of the -SiO(3) units. The intense Raman band at 3479 cm(-1) is ascribed to the stretching vibration of the OH units. The Raman band at 3250 cm(-1) is evidence for water in the structure.


Cryo-fluorescence micro-optical sectioning tomography for volumetric imaging of various whole organs with subcellular resolution.
Optical visualization of complex microstructures in the entire organ is essential for biomedical research. However, the existing methods fail to accurately acquire the detailed microstructures of whole organs with good morphological and biochemical preservation. This study proposes a cryo-fluorescence micro-optical sectioning tomography (cryo-fMOST) to image whole organs in three dimensions (3D) with submicron resolution. The system comprises a line-illumination microscope module, cryo-microtome, three-stage refrigeration module, and heat insulation device. To demonstrate the imaging capacity and wide applicability of the system, we imaged and reconstructed various organs of mice in 3D, including the healthy tongue, kidney, and brain, as well as the infarcted heart. More importantly, imaged brain slices were performed sugar phosphates determination and fluorescence in situ hybridization imaging to verify the compatibility of multi-omics measurements. The results demonstrated that cryo-fMOST is capable of acquiring high-resolution morphological details of various whole organs and may be potentially useful for spatial multi-omics.


Fasciitis ossificans: imaging features, histology, and differential diagnosis.
To describe the imaging features of fasciitis ossificans and its histopathological features. Using a word search of existing pathology reports at the Mayo Clinic, six cases of fasciitis ossificans were identified. The clinical history, histology, and available imaging of the affected area were reviewed. Imaging consisted of radiographs, mammograms, ultrasound images, bone scintigraphs, computed tomography (CT), and magnetic resonance imaging (MRI) images. All cases demonstrated a soft-tissue mass. The characteristic MRI appearance was a T2 hyperintense enhancing mass with surrounding soft-tissue oedema. Peripheral calcifications were seen on radiographs, CT, and/or ultrasound. Histological sections showed distinct zonation, with nodular fasciitis-like zones of myofibroblastic proliferation, which merged with osteoblasts that rim the ill-defined trabeculae of woven bone and became continuous with the mature lamellar bone surrounded by a thin layer of compressed fibrous tissue. Imaging features of fasciitis ossificans are that of an enhancing soft-tissue mass located within a fascial plane with prominent surrounding oedema and mature peripheral calcification. Imaging and histology are that of myositis ossificans but occurring within the fascia. It is important that radiologists are aware of the diagnosis of fasciitis ossificans and appreciate its similarity to myositis ossificans. This is particularly important in anatomical locations with fascias but no muscle. Given the radiographic and histological overlap between these entities, nomenclature that encompasses both could be considered in the future.


Crystal structure of bis-(μ-3-nitro-benzoato)-κ3O,O':O;κ3O:O,O'-bis-[bis-(3-cyano-pyridine-κN1)(3-nitro-benzoato-κ2O,O')cadmium].
The asymmetric unit of the title compound, [Cd2(C7H4NO4)4(C6H4N2)4], contains one CdII atom, two 3-nitro-benzoate (NB) anions and two 3-cyano-pyridine (CPy) ligands. The two CPy ligands act as monodentate N(pyridine)-bonding ligands, while the two NB anions act as bidentate ligands through the carboxyl-ate O atoms. The centrosymmetric dinuclear complex is generated by application of inversion symmetry, whereby the CdII atoms are bridged by the carboxyl-ate O atoms of two symmetry-related NB anions, thus completing the distorted N2O5 penta-gonal-bipyramidal coordination sphere of each CdII atom. The benzene and pyridine rings are oriented at dihedral angles of 10.02 (7) and 5.76 (9)°, respectively. In the crystal, C-H⋯N hydrogen bonds link the mol-ecules, enclosing R22(26) ring motifs, in which they are further linked via C-H⋯O hydrogen bonds, resulting in a three-dimensional network. In addition, π-π stacking inter-actions between parallel benzene rings and between parallel pyridine rings of adjacent mol-ecules [shortest centroid-to-centroid distances = 3.885 (1) and 3.712 (1) Å, respectively], as well as a weak C-H⋯π inter-action, may further stabilize the crystal structure.


Pressurized intraperitoneal aerosol chemotherapy (PIPAC) for rare gynecologic indications: peritoneal metastases from breast and endometrial cancer.
Peritoneal metastasis (PM) in patients with breast (BC) and endometrial cancer (EC) is rare and treatment options are limited. Pressurized IntraPeritoneal Aerosol Chemotherapy (PIPAC) has demonstrated efficacy against PM from various cancers, but its efficacy in BC/EC patients is unknown. Retrospective cohort study of patients with PM from BC/EC undergoing PIPAC with doxorubicin 1.5 mg/m2 and cisplatin 7.5 mg/m2. Data were collected within an international prospective PIPAC registry. Study outcomes were microscopic tumor regression grade (TRG), survival, adverse events (CTCAE), and quality of life (QoL). 150 PIPAC procedures in 44 patients (BC/EC = 28/16; mean age 58.8 ± 10.1 and 63.2 ± 10.1 years, respectively) were analyzed. The mean number of PIPACs per patient was 3 (range 0-9) and 3.5 (range 0-10), respectively. Primary/secondary non-access occurred in 4/3 of 150 (5%) procedures. PIPAC induced objective tumor regression as demonstrated by repetitive PM biopsies in 73% (32/44) of patients. Peri- and postoperative CTCAE grade 3 and 4 complications were observed in 12/150 (8%) of procedures. No grade 5 event was observed. After a median follow up of 5.7 (IQR 2.7-13.0) months, overall median survival was 19.6 (95% CI: 7.8-31.5) months (from first PIPAC). QoL indicators (general health, nausea, fatigue, constipation, pain, dyspnea, social, cognitive, emotional, and physical functioning) all improved or were maintained throughout PIPAC treatments. Repetitive intraperitoneal chemotherapy with PIPAC is feasible and safe in patients with PM from BC and EC. PIPAC induces significant histological regression of PM while maintaining QoL.


Lomefloxacin: microbiologic assessment and unique properties.
In comparative studies, lomefloxacin, a new difluorinated quinolone, exhibits broad antibacterial activity in vitro, similar or superior to that of other quinolones (enoxacin, ofloxacin, pipemidic acid, nalidixic acid, and norfloxacin) but less than that of ciprofloxacin. Lomefloxacin inhibited Neisseria gonorrhoeae, Moraxella (Branhamella) catarrhalis, Haemophilus influenzae, Pseudomonas aeruginosa, Staphylococcus aureus, and the majority of aerobic gram-negative rods, including nosocomial isolates, at concentrations readily achievable in biologic fluids and tissues. Lomefloxacin was less active against obligate anaerobes and streptococci. Organisms resistant to methicillin, penicillin, or the aminoglycosides were susceptible to lomefloxacin. No significant lomefloxacin resistance was identified in 18 countries in which in vitro studies were conducted, with the exception of a small number of strains tested in France. The frequency with which spontaneous single-step resistance to lomefloxacin develops in vitro is low.


Temporal-spatial expression and transcriptional regulation of alpha7 nicotinic acetylcholine receptor by thyroid transcription factor-1 and early growth response factor-1 during murine lung development.
Nicotinic acetylcholine receptors are ligand-gated ion channels formed by five homologous subunits that are involved in processes including signal transduction, proliferation, and apoptosis. The developmental role of these receptors, however, is unclear. In the present investigation, alpha(7) nicotinic acetylcholine receptor expression was assessed by immunohistochemistry in mouse lungs from embryonic day (E)13.5 to postnatal day (PN)20. Transcriptional mechanisms that regulate alpha(7) were assessed by the transfection of murine bronchiolar cells with a reporter containing 1.1 kb of the mouse alpha(7) promoter, TTF-1, and Egr-1. alpha(7) was initially detected at E13.5 in pulmonary mesenchymal cells and in the epithelium of the primitive tubules at E15.5. From E18.5 to PN1, alpha(7) was expressed in conducting airway and saccule epithelial cells. By PN10, expression was observed in the peripheral epithelium and on luminal membranes of bronchiolar epithelial cells in the proximal lung, a pattern that continued through PN20. From E15.5 to PN20, type II alveolar cells expressed both prosurfactant protein C and alpha(7). From E18.5 to PN20, Clara cells in the bronchiolar epithelium co-expressed Clara cell secretory protein and alpha(7). TTF-1 dose-dependently activated alpha(7) transcription in vitro by binding specific TTF-1 regulatory elements in the mouse alpha(7) promoter. Furthermore, alpha(7) was not detected in TTF-1-null mice and markedly increased in TTF-1-overexpressing mice. Conversely, Egr-1 inhibited alpha(7) expression. Temporal-spatial alpha(7) expression supports the concept that these receptors function during normal pulmonary morphogenesis. A model is also supported whereby alpha(7) is induced by the essential pulmonary transcription factor TTF-1 and suppressed by Egr-1 during pulmonary development.


Oral L-arginine and vitamins E and C improve endothelial function in women with type 2 diabetes.
Endothelial dilator function is impaired in people with type 2 diabetes mellitus (T2DM). Prior research indicates that this can be improved with intravenous administration of ascorbate or L-arginine, but whether these agents have this effect when administered by the clinically practical oral route is unknown. To investigate this question, 10 premenopausal women with T2DM and 10 healthy, premenopausal, non-diabetic women received, in random sequence, a 1-week administration of oral L-arginine (9 g daily) or vitamins E (1800 mg) and C (1000 mg) with an intervening 1-week washout period. Flow-mediated brachial artery dilation (FMD) was measured by ultrasonography and forearm blood flow was measured by plethysmography before and following blood pressure cuff-induced forearm ischemia before and after each week of treatment. At baseline, the women with T2DM had lesser FMD responses (0.028 +/- 0.006 cm vs 0.056 +/- 0.008 cm, p < 0.05). Post-ischemic forearm hyperemia was reduced at baseline in T2DM compared with controls (16.4 +/- 1.8 vs 26.0 +/- 1.4 ml 100 ml(-1) min(-1), p < 0.05). Administration of L-arginine caused a 50 +/- 12% increase in FMD in T2DM (p < 0.05) and raised post-ischemic forearm blood flow by 29 +/- 8% (p < 0.05). No significant changes were seen in controls. Administration of vitamins E and C in women with T2DM produced an increase in the brachial artery diameter response of 79 +/- 15% (p < 0.05), but did not significantly increase the hyperemic blood flow response (p = NS). No significant changes in the responses of controls from pre to post vitamin administration were observed. We concluded that administration of two types of oral agents improved measures of endothelial function in people with T2DM.


Primary small-cell carcinoma of the breast: a case report.
We report a case of small-cell carcinoma of the breast. The patient was a 54-year-old Japanese woman with a left breast mass. Mammography revealed a 1.2-cm microlobulated mass with partially ill-defined borders. Ultrasonography disclosed a hypoechoic mass measuring 1.8 × 1.2 × 1.2 cm with heterogeneous internal echoes and partially ill-defined borders. The shape of the mass was round and taller than it was wide in the orthogonal section to the longest axis of the mass. These imaging findings suggested a malignant tumor. Fine-needle aspiration cytology of the tumor also suggested a ductal carcinoma. Breast-conserving surgery was performed with axillary sentinel lymph node biopsy. A diagnosis of small-cell carcinoma of the breast was made based on the histologic and immunohistochemical findings. This rare breast tumor has been reported to be aggressive and associated with a poor prognosis; however, our patient is currently well and has had no clinical recurrence of the disease after 5 years of follow-up without radiotherapy or chemotherapy. Therefore, the prognosis may be better if the tumor is detected early and there is an absence of lymph node metastasis.


A fan-attached jacket worn in an environment exceeding body temperature suppresses an increase in core temperature.
We examined whether blowing hot air above body temperature under work clothing may suppress core temperature. Nine Japanese men engaged in two 30-min bicycle ergometer sessions at a workload of 40% VO2max at 40 °C and 50% relative humidity. The experiment was conducted without wearing any cooling apparatus (CON), wearing a cooling vest that circulated 10.0 °C water (VEST), and wearing a fan-attached jacket that transferred ambient air underneath the jacket at a rate of 30 L/s (FAN). The VEST and FAN conditions suppressed the increases of rectal temperature (CON, VEST, FAN; 38.01 ± 0.19 °C, 37.72 ± 0.12 °C (p = 0.0076), 37.54 ± 0.19 °C (p = 0.0023), respectively), esophageal temperature (38.22 ± 0.30 °C, 37.55 ± 0.18 °C (p = 0.0039), 37.54 ± 0.21 °C (p = 0.0039), respectively), and heart rate (157.3 ± 9.8 bpm, 136.9 ± 8.9 bpm, (p = 0.0042), 137.5 ± 6.5 bpm (p = 0.0023), respectively). Two conditions also reduced the estimated amount of sweating and improved various subjective evaluations. Even in the 40 °C and 50% relative humidity environment, we may recommend wearing a fan-attached jacket because the heat dissipation through evaporation exceeded the heat convection from the hot ambient air.


Correction to: Mucormycosis in hospitalized patients at a tertiary care center in Lebanon: a case series.
The spelling of the name "Ali Bazarbachi" was incorrect.


Comparison of an in-house method and the commercial Sepsityper™ kit for bacterial identification directly from positive blood culture broths by matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry.
The identification of bacteria directly from positive blood cultures using matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry (MALDI-TOF MS) is a new challenge to microbiologists. However, the protocols previously described are often difficult to implement in routine and comparisons are not always possible due to the variability of interpretative criteria. This study evaluated the analytical and practical performances of an in-house (IH) method, adapted from previous protocols, and the Sepsityper™ kit (Bruker Daltonics, Bremen, Germany). Positive blood cultures from 63 different patients were prospectively evaluated by both methods. To enhance the sensitivity of these methods, lowered cut-offs were assessed and validated on 66 additional samples. The IH method produced 86.4% and 73.7% correct genus and species identifications, respectively, when using the lowered cut-offs of 1.4 and 1.6 for correct genus and species identifications. The Sepsityper™ kit showed similar results (78.0% and 68.4% correct genus and species identification, respectively). However, the IH method is ten-fold less expensive than the commercial option (0.72 vs. 7.45 <euro>/analysis) and its turnaround time is approximately 20 min versus the nearly 40 min required for the Sepsityper™ kit, which includes an extraction step. Finally, the IH method was introduced twice-daily in our routine practice.


Comparison of abilities of various interpretations of bio-electrical impedance to predict reference method body composition assessment.
Body composition was assessed in 28 normal men and women using three established reference methods (deuterium dilution for total body water, and densitometry and dual-energy X-ray absorptiometry for body fat and fat-free mass), the four- and three-component models based on these methods, and eight different predictions of the same whole-body bio-electrical impedance measurement. The comparative value of each prediction was assessed using bias and 95% limits of agreement against all reference methods. The magnitude of bias between each interpretation of whole body impedance and each reference method estimate was found to vary considerably, but in general this was consistent over the range studied. However, with few exceptions, magnitudes of the limits of agreement were considerable. For example, a majority of values of 95% limits of agreement in predicting total body water reached bias+/-3 kg or above, and all values in predicting fat (% body weight) for all subjects reached bias+/-6%, and for most purposes these could well be unacceptable. Although in absolute terms bias and limits of agreement for fat-free mass were the same as for body fat, estimates of fat-free mass were relatively more acceptable because fat-free mass was (as is the usual case) the larger of these two body components.


Clearance of pneumococcal organisms after repair of injured spleens.
Although the safety of splenic preservation has been demonstrated in a small number of cases, the follow-up in these cases has been too short to determine whether the patients are adequately protected from sepsis. The purpose of this study was to test the ability of the spleen to clear pneumococcal organisms after repairing lacerations with a surface hemostatic agent. Three groups of 20 New Zealand white rabbits were studied. Group I animals received sham operations, group II animals were given total splenectomies, and group III animals had splenic lacerations repaired with Collastat (American Medical Products Corp., Freehold, New Jersey), a local hemostatic agent. Three months later, when challenged with intracardiac Streptococcus pneumoni type III, the animals without spleens failed to clear pneumococcal organisms. This spleen-intact and spleen-repaired animals, however, both cleared pneumococcal organisms in a normal fashion.


Latencies of calcium elevation and depolarization in Limulus ventral photoreceptors injected with GDP-beta S.
We have used confocal fluorescence microscopy and fluorescent calcium indicators to investigate the relationship between light-induced elevation of intracellular calcium ion concentration (Cai) and depolarization in small volumes of cytosol close to the microvillar plasma membrane of the ventral photoreceptor of Limulus polyphemus. We prolonged the latency of the light response by treatment of cells with hydroxylamine and injection of the G-protein blocker, guanosine-5'-O-(2-thiodiphosphate (GDP-beta S). Such treatment increased the latency of the cell's response from approximately 20 to 50 ms. In both treated and untreated cells we observed a close correlation between the times at which we first detected the electrical response and the elevation of Cai. We obtained 18 out of 54 and 12 out of 22 recordings, in untreated and treated cells respectively, for which the elevation of Cai was detected simultaneous with, or 1-4 ms prior to, the electrical response to light. The prolonged latent period exhibited by treated cells may make possible future investigation of the effects on the initial response to light of local photolytic release of caged compounds at the microvillar membrane.


Amino-Acid-Induced Circular Polarized Luminescence in One-Dimensional Manganese(II) Halide Hybrid.
Circular polarized luminescence (CPL)-active materials attract great attentions owing to their widely applications in 3D optical displays and encrypted transmission. Inspired by the strategies adopted in perovskite based CPL materials, herein, CPL-active hybrids (D)- and (L)-(tert-butyl prolinate)MnCl3 were successfully prepared by assembling chiral D/L tert-butyl prolinate with manganese (II) chloride. Single crystal structures show the as-formed hybrids possess one-dimensional (1D) structure containing linear chains of face-sharing MnCl6 octahedral surrounded by prolinate cations. The 1D Mn(II) hybrids display strong red emission peaked at 646 nm with PLQY of 67.1 % and 57.2 % for d-type and l-type, respectively, representing the highest PLQY for 1D MnII hybrids. Interestingly, the 1D Mn(II) hybrids exhibit prominent circular dichroism (CD) signals and remarkable CPL activity with the dissymmetry factor g of 6.1*10-3 and -6.3*10-3 from 550 to 800 nm for (D)- and (L)-(tert-butyl prolinate)MnCl3 , respectively, owing to the existence of chiral cations. It is worthy noted the obtained g represents the highest value for non-lead organic-inorganic hybrids.


A snap-shot of orthokeratology lens rinsing habits: An online survey in a major eye hospital in Shanghai.
A hospital-based survey to explore lens rinsing habits of orthokeratology (ortho-k) patients, including the solution used for rinsing. An anonymous survey, via an online questionnaire, was administered to ortho-k lens wearers or their parents when they attended Fudan University Eye and ENT Hospital for routine follow-up visits. Three major issues were investigated: basic demographics, lens rinsing solution used, and specific rinsing habits. Relationships between demographics and rinsing solution used and rinsing behaviours were explored. Of the total of 541 online questionnaires delivered, 296 were completed and returned. The respondents were the person responsible for lens care, either the ortho-k users themselves, or their parents. The median age of the lens wearers was 12.0 years (range: 8-22 years), and the median period of ortho-k lens wear was 18.0 months (range: 1-109 months). The most common rinsing solutions reported were a particular brand of multidose preserved saline (28.4%), cooled boiled water (23.3%), and non-preserved multidose saline (16.2%). Overall, the main reason for choice of rinsing solution was recommendation of eye care practitioners (90.2%). The respondents with a longer lens wear history tended to use commercially available bottled water, drinking water, or cooled boiled water rather than non-preserved saline (unidose or multidose) (p ≤ 0.009) and were more likely to use the rinsing solution to soak their lenses (p = 0.009). Lens wearers who rinsed the lenses themselves tended to use tap water compared to those whose lenses were handled by parents (p = 0.040). Inappropriate choice and usage of rinsing solution appeared to be a common issue among ortho-k lens wearers in Shanghai, China. The problem was more pronounced in long-term wearers and in those who performed rinsing themselves.


Disability: how successful are we in determining disability?
Individuals who have severe back pain have trouble bending, may not be able to put on their shoes, have difficulty in ambulation, and may not be able to clean their own houses. What distinguishes these individuals with low back pain from those afflicted with other conditions (i.e., congestive heart failure, metastatic cancer, symptomatic acquired immune deficiency syndrome) is prognosis. Back pain essentially never shortens life. Acute back pain has a very benign prognosis, with more than 90% of the individuals returning to functional status equivalent to their baseline status within 3 months of the onset of pain. Patients who have chronic back pain have a significantly worse prognosis, but most cohort studies show that substantial numbers of chronic back pain patients improve over time with supportive therapy.


Human immunodeficiency virus type-1 (HIV-1) Pr55gag virus-like particles are potent activators of human monocytes.
Human immunodeficiency virus type-1 (HIV-1) Pr55(Gag) virus-like particles (VLP) represent an interesting HIV vaccine component since they stimulate strong humoral and cellular immune responses. We demonstrated that VLP expressed by recombinant baculoviruses activate human PBMC to release pro-inflammatory (lL-6, TNF-alpha), anti-inflammatory (IL-10) and Th1-polarizing (IFN-gamma) cytokines as well as GM-CSF and MIP-1alpha in a dose-and time-dependent manner. Herein, residual baculoviruses within the VLP preparations showed no or minor effects. Monocytes could be identified as a main target for VLP to induce cytokine production. Furthermore, VLP-induced monocyte activation was shown by upregulation of molecules involved in antigen presentation (MHC II, CD80, CD86) and cell adhesion (CD54). Exposure of VLP to serum inactivates its capacity to stimulate cytokine production. In summary, these investigations establish VLP as strong activators of PBMC and monocytes therein, potently enhancing their functionality and potency to promote an efficient immune response. This capacity makes VLP an interesting component of combination vaccines.


Improving the climate safety of workers in Almería-type greenhouses in Spain by predicting the periods when they are most likely to suffer thermal stress.
The humidex and wind-chill indices were used to determine the periods in which labourers working in Almería-type greenhouses in southeastern Spain are most likely to suffer conditions able to induce heat and cold stress. Over 500,000 pieces of data for wet and dry bulb temperatures and relative humidity, recorded over a period in excess of five years by a weather station located in an Almería-type greenhouse containing a grass crop, were used in the calculation of these indices. The wind-chill index results showed cold stress never to be a problem, but the humidex index results showed that conditions under which heat stress could develop were common in the warmer months. A clock diagram was produced showing the hours when heat stress is likely to occur in each month of the year. This information could be used to improve the conditions of labourers working in this type of greenhouse; some ways of reducing their exposure to heat stress-inducing conditions are discussed.


BLINCAR: a reusable bioluminescent and Cas9-based genetic toolset for repeatedly modifying wild-type Scheffersomyces stipitis.
Scheffersomyces stipitis is a yeast that robustly ferments the 5-carbon sugar xylose, making the yeast a valuable candidate for lignocellulosic ethanol fermentation. However, the non-canonical codon usage of S. stipitis is an obstacle for implementing molecular tools that were developed for other yeast species, thereby limiting the molecular toolset available for S. stipitis. Here, we developed a series of molecular tools for S. stipitis including BLINCAR, a Bio-Luminescent Indicator that is Nullified by Cas9-Actuated Recombination, which can be used repeatedly to add different exogenous DNA payloads to the wild-type S. stipitis genome or used repeatedly to remove multiple native S. stipitis genes from the wild-type genome. Through the use of BLINCAR tools, one first produces antibiotic-resistant, bioluminescent colonies of S. stipitis whose bioluminescence highlights those clones that have been genetically modified; then second, once candidate clones have been confirmed, one uses a transient Cas9-producing plasmid to nullify the antibiotic resistance and bioluminescent markers from the prior introduction, thereby producing non-bioluminescent colonies that highlight those clones which have been re-sensitized to the antibiotic and are therefore susceptible to another round of BLINCAR implementation. IMPORTANCE Cellulose and hemicellulose that comprise a large portion of sawdust, leaves, and grass can be valuable sources of fermentable sugars for ethanol production. However, some of the sugars liberated from hemicellulose (like xylose) are not easily fermented using conventional glucose-fermenting yeast like Saccharomyces cerevisiae, so engineering robust xylose-fermenting yeast that is not inhibited by other components liberated from cellulose/hemicellulose will be important for maximizing yield and making lignocellulosic ethanol fermentation cost efficient. The yeast Scheffersomyces stipitis is one such yeast that can ferment xylose; however, it possesses several barriers to genetic manipulation. It is difficult to transform, has only a few antibiotic resistance markers, and uses an alternative genetic code from most other organisms. We developed a genetic toolset for S. stipitis that lowers these barriers and allows a user to deliver and/or delete multiple genetic elements to/from the wild-type genome, thereby expanding S. stipitis's potential.


Robotic-assisted apical lateral suspension for advanced pelvic organ prolapse: surgical technique and perioperative outcomes.
Abdominal sacral hystero-cervicopexy (ASC) is the gold standard for the treatment of apical prolapse, but requires advanced laparoscopic skills and exposes to potentially life-threatening complications. Lateral apical suspension to the abdominal wall with mesh is a feasible alternative of ASC where robotic assistance may offer specific advantages. We here describe the surgical technique and the short-term outcomes of robotic-assisted lateral apical suspension (R-ALS) with the use of a titan-covered T-shaped mesh. Forty consecutive patients with stage III or IV symptomatic anterior and apical pelvic organ prolapse underwent R-ALS between September 2014 and September 2015. R-ALS was completed without complications in all cases with a mean operative time of 117 ± 26 min. From a technical standpoint, robotic assistance allowed for an extremely reproducible technique, with a swift learning curve and consistent length of the surgical steps. The procedure was extremely well tolerated and resulted in complete resolution of POP-associated symptoms and in improvements of POP- and incontinence-related quality-of-life scores (PQOL and IIQ7) at 1 month from surgery. R-ALS is feasible, safe, well-tolerated and effective at a short-term follow-up. R-ALS may represent an effective and simple alternative to abdominal sacral hystero-cervicopexy for the treatment of high-grade apical and anterior POP, avoiding the challenges of sacral mesh fixation. Robotic assistance helps achieving optimally tailored anatomic reconstruction, allowing seamless deep pelvic dissection and suturing.


Heart rate variability increases in elderly depressed patients who respond to electroconvulsive therapy.
To evaluate the parasympathetic modulation in elderly inpatients with major depressive disorder (MDD) before and after electroconvulsive therapy (ECT) using both linear and nonlinear methods of heart rate variability (HRV) analysis. A measure of local dimensional complexity (pointwise correlation dimension, PD2), as well as spectral analysis measures (LF, low-frequency range; HF, high-frequency range) were calculated for the heart rate time series of 10 elderly inpatients with MDD (70+/-7 years) before and after ECT. Hamilton Depression Rating Scale (HAM-D) was evaluated concomitantly. Only the responders to ECT (n=7; >or=50% reduction in HAM-D) exhibited a significant increase in PD2 (P=.0035), which showed a tendency towards a correlation with symptom improvement (r=.73, P=.06). Spectral analysis measures did not show a significant difference after ECT. Elderly patients with MDD, who respond to ECT, might show increased vagal modulation. Since nonlinear HRV measures have been shown to be reduced by aging, similar to cholinergic deficit, they might shed light on the increased risk for cardiac mortality in depression.


Upregulation of myeloperoxidase in patients with opticospinal multiple sclerosis: positive correlation with disease severity.
To clarify the role of myeloperoxidase (MPO) in multiple sclerosis (MS), we measured serum MPO levels in 86 Japanese patients with relapsing remitting MS, 47 with opticospinal MS (OSMS) and 39 with conventional MS (CMS), and 85 healthy subjects by sandwich enzyme immunoassays and analyzed relationships with clinical features. We found a significant increase in serum MPO in OSMS patients at relapse and remission, and in CMS patients at remission compared with controls. By logistic regression analysis, the clinical variable associated with high level of MPO at remission in OSMS patients (higher than the mean+/-2 S.D. of healthy controls) was only Kurtzke's Expanded Disability Status Scale (EDSS) score in blood sampling (p=0.0245); that is, a greater EDSS scores in the high MPO group, whereas in CMS none were associated. The results of our study suggest that MPO levels in remission are related with severe tissue destruction in OSMS.


Malignant rhabdoid tumor of soft tissues: a cytopathological and immunohistochemical study.
We describe the cytopathological and immunohistochemical features of a primary malignant rhabdoid tumor (MRT) located in the left forehead region of an 8-mo-old female. The cardinal characteristics are hyaline cytoplasmic inclusions, rounded or bean-shaped nucleus, vesicular chromatin, central prominent nucleolus, and poorly defined cytoplasmic profile. Both the cytoplasm and the hyaline inclusion are vimentin and keratin positive, but negative with desmin, S-100 protein, neuron-specific enolase, (NSE) myoglobin, leukocyte common antigen (LCA), and alpha-1-antichymotrypsin. This immunohistochemical pattern, although not exclusive to MRT, is fairly typical of MRT, and when it is assessed in conjunction with cytopathological findings and clinical data permits a confident diagnosis of MRT. We emphasize the utility of immunohistochemical techniques in making the differential diagnosis of MRT against other childhood round-cell tumors.


Measurement of Michaelis constant for human erythrocyte transketolase and thiamin diphosphate.
Human erythrocyte transketolase could be resolved from thiamin diphosphate (TDP) by acidification of the ammonium sulfate precipitate to pH 3.5, but not by other tested procedures. Resolution was 98% by chemical measurement of residual thiamin and 95% by residual enzyme activity. Reconstitution of the resolved preparation by incubation with TDP was dependent upon TDP concentration, duration, temperature, and the presence of dithiothreitol. At low TDP concentrations, 1 h was required for maximum activation; kinetic analysis then yielded an apparent Km value for TDP of 65 nM (SD 14 nM) from 100 erythrocyte lysates and similar values for reconstituted resolved preparations previously purified 400-fold and 10,000-fold. Velocity data obtained by transketolase assays in which the TDP was added to resolved preparations simultaneously with substrates yielded an apparent Km value for TDP of 2.3 microM (SD 1.6 microM) from 114 erythrocyte lysates and similar values for purified preparations. The recovery of activity following resolution and reconstitution ranged from 21 to 60% from lysates and 38 to 70% from purified preparations. Residual ammonium sulfate up to 4.9 mM decreased the apparent Km value for TDP, while a concentration of 11.3 mM increased the value in a manner competitive with TDP and with an apparent Ki value of 2.3 mM. The spectrophotometric assay of transketolase activity was greatly affected by storage of frozen solutions of the substrate ribose 5-phosphate.


A role for Akt in the rapid regulation of inflammatory and apoptotic pathways in mouse bladder.
Akt is linked to both inflammatory and neoplastic pathways. Akt activation is dependent on the phosphatidylinositol-3 kinase (PI3K) signaling pathways. Upon phosphorylation by PI3K, Akt can phosphorylate nuclear factor kappa B (NF-kappaB) and members of the forkhead family of transcription factors, which includes AFX. Our goal is to examine the effect of Escherichia coli lipopolysaccharide (LPS) on early cellular signaling in inflammatory (NF-kappaB) and apoptotic pathways (AFX) in a mouse-bladder model and in T-24 urothelial cancer cells. Female C57BL/6 mice were given an intraperitoneal (IP) injection of LPS or LPS free water and sacrificed 0-120 minutes later. Bladders were harvested, and immunohistochemistry (IHC) and/or immunoblotting performed using antibodies to PI3K, inhibitor kappa B-alpha (IkappaB-alpha), and total and phosphorylated Akt, NF-kappaB and AFX. Levels of IkappaB-alpha and total and phosphorylated Akt and NF-kappaB were determined in T-24 cells treated with LPS for 0-120 minutes. Bladders and T-24 cells were treated with PI3K inhibitors in some experiments. Protein amounts in different samples were normalized to immunoreactive actin. Phosphorylated and non-phosphorylated species of Akt, NF-kappaB, and AFX were localized to the urothelium. IP LPS injection rapidly (within 30 minutes) increased Akt phosphorylation. IP LPS injection decreased IkappaB-alpha levels, and increased NF-kappaB and AFX phosphorylation. Wortmannin effectively blocked phosphorylation of Akt in LPS-treated mice, and also reduced phosphorylation of AFX and, to a lesser extent, NF-kappaB. After treatment with LPS, Akt and NF-kappaB phosphorylation was rapidly increased in T-24 cells. Akt phosphorylation, and to a lesser extent NF-kappaB phosphorylation, were blocked by LY-294,002. LPS/PI3K/Akt is a cellular signaling pathway which rapidly activates downstream pathways of inflammation and neoplasia in bladder urothelium.


EGFR L2 domain mutation is not correlated with resistance to cetuximab in metastatic colorectal cancer patients.
The KRAS mutation has been associated with resistance to cetuximab, an anti-epidermal growth factor receptor (EGFR) monoclonal antibody, in metastatic colorectal cancer (mCRC). However, the predictive biomarkers of cetuximab resistance in KRAS wild-type mCRC remain unknown except BRAF, NRAS, and PIK3CA exon 20. The objective of the study is to study the impact of EGFR L2 mutations on resistance to cetuximab in KRAS wild-type patients. A total of 247 mCRC patients were screened for KRAS status at the National Cancer Center Hospital between September 2008 and April 2010. We analyzed the EGFR L2 domain mutation status in KRAS wild type and in the patients treated with cetuximab-based therapy. There were 136 patients with wild-type KRAS (55%). Sixty-five patients were analyzed for the L2 domain mutation status, and all patients received cetuximab-based therapy. One patient who had a mutation at exon 9 showed a partial response to cetuximab plus irinotecan. Mutation of the EGFR L2 domain was analyzed in mCRC patients. Our findings do not provide sufficient evidence that EGFR L2 domain mutation is correlated with resistance to cetuximab.


Characterization of canine triiodothyronine (T3) autoantibodies and their effect on total T3 in canine serum.
A study of 3,5,3'-L-triiodothyronine autoantibody (T3 AA) in 18 dogs revealed an average apparent affinity constant for T3 of 2.24 +/- 1.78 X 10(10) M-1, an average T3 binding capacity of 639.3 +/- 666.5 ng/dl and a low thyroxine (T4) cross-reactivity (less than 1%) in all samples tested. A valid radioimmunoassay (RIA) procedure which involved heat treatment of samples for 1 hr at 70 degrees C and assay on Sephadex minicolumns was developed for measuring T3 in the presence of T3 AA. Total T3 was elevated (mean = 374.8 +/- 158.4 ng/dl) in samples in which T4 was in the normal canine range, but T3 was lower (mean = 96.1 +/- 63.3 ng/dl) in samples with T4 values in the hypothyroid range. For each sample the concentration of T3 not bound by T3 AA was calculated from the total T3 concentration, the affinity constant, and the binding capacity. In dogs with normal total T4 concentrations the average calculated T3 not bound by T3 AA was 147.2 +/- 144.4 ng/dl while in dogs with low total T4 the value was 15.7 +/- 26.3 ng/dl (normal canine range is 45-150 ng/dl). Canine samples containing T3 AA were compared to serum from three rabbits actively immunized against T3 to provide anti-T3 for commercial RIA. The rabbit T3-antisera had an average T3 affinity constant similar to those of the canine samples (1.57 X 10(10) M-1), but had average titer, T3 binding capacity, and total T3 values more than 10-fold higher. Our findings indicate that, in dogs with serum containing T3 AA and normal total T4 concentrations, a compensatory mechanism appears to exist to maintain non-T3 AA bound T3 within the range of normal total T3. This compensatory mechanism does not operate in those dogs with insufficient thyroid activity to maintain normal total T4 values.


Is the Fab-fragment from monoclonal rheumatoid IgM flexible? A spin label dynamics study.
A comparative study of the Fab- and Fab-RF-fragments in the molecules of monoclonal IgM and IgM-RF, respectively, was performed by the spin label method. The spin label, 2,2,6,6-tetramethyl-4-dichloro-triazinylaminopiperidine-1-oxyl, was introduced into the peptide part of the protein. On the basis of the data on the temperature-viscosity dependences of the EPR spectral parameters of the resultant spin-labeled proteins, the rotational correlation time tau of the spin carrier was determined. It turned out that the reduced to normal conditions tau values for the molecules of the Fab- and Fab-RF-fragments were 21+/-2 and 11+/-1 ns, respectively. Analysis of the resultant data provides sufficient grounds for assuming that such a sharp decrease in the tau value for the molecule of the Fab-RF fragment is due to local flexibility of its globular structure, which, in turn, can determine the specific features of the IgM-RF functioning as an autoantibody.


Novel Venetin-1 nanoparticle from earthworm coelomic fluid as a promising agent for the treatment of non-small cell lung cancer.
The present research shows the antitumor activity of a protein-polysaccharide complex Venetin-1 obtained from the coelomic fluid of Dendrobaena veneta earthworms against A549 cancer cells. The investigations are a continuation of experiments on the antitumor activity of coelomic fluid obtained from this species. The Venetin-1 nanoparticle was obtained after thermal treatment of the coelomic fluid, separation from coelomocytes, filtration, and lyophilization. The preparation showed a selective effect on cancer cells, whereas normal cells were unaffected. Venetin-1 was effective against the lung cancer cells at doses of 31.3 and 62.5 µg/ml, and the results were imaged using light microscopy and scanning electron microscopy (SEM). The cells died mainly via the apoptosis pathway. Necrotic cells appeared sporadically in the microscopic view. SEM imaging revealed complete destruction of the A549 cells after the incubation with Venetin-1. The atomic force microscopy (AFM) analyses showed changes in the topography, peak force error images, and Young's modulus (elasticity) of the A549 cells after the incubation with Venetin-1. The transmission electron cryomicroscopy (Cryo-TEM) analysis indicated a polymeric nature of the analyzed preparation. The samples of Venetin-1 showed a very homogeneous size profile with the microparticle size of approximately 58.23 nm. A significant decrease in Venetin-1 binding to sphingomyelin was observed. Venetin-1 lost its pore-forming activity or deactivation of the pore-forming activity occurred. This confirms the absence of hemolytic capacity of Venetin-1 towards red blood cells. The conducted analyses show the suitability of the obtained complex for biomedical research. The next step will consist in analyses of the effect of Venetin-1 on the immune system in mice.


Nascent alt-protein chemoproteomics reveals a pre-60S assembly checkpoint inhibitor.
Many unannotated microproteins and alternative proteins (alt-proteins) are coencoded with canonical proteins, but few of their functions are known. Motivated by the hypothesis that alt-proteins undergoing regulated synthesis could play important cellular roles, we developed a chemoproteomic pipeline to identify nascent alt-proteins in human cells. We identified 22 actively translated alt-proteins or N-terminal extensions, one of which is post-transcriptionally upregulated by DNA damage stress. We further defined a nucleolar, cell-cycle-regulated alt-protein that negatively regulates assembly of the pre-60S ribosomal subunit (MINAS-60). Depletion of MINAS-60 increases the amount of cytoplasmic 60S ribosomal subunit, upregulating global protein synthesis and cell proliferation. Mechanistically, MINAS-60 represses the rate of late-stage pre-60S assembly and export to the cytoplasm. Together, these results implicate MINAS-60 as a potential checkpoint inhibitor of pre-60S assembly and demonstrate that chemoproteomics enables hypothesis generation for uncharacterized alt-proteins.


Searching in clutter: Visual behavior and performance of expert action video game players.
Searching for targets among distractors in visual scenes can be more difficult due to the presence of clutter. However, studies in various domains have shown differentiated effects according to the expertise of the searcher. The present study extended these findings to the domain of action video games expertise. 58 participants, split in 2 groups (action video game players and non-action video game players) searched for targets in visual scenes under two clutter conditions (uncluttered and high clutter). Reaction times and accuracy served as measures of performance, and the visual behavior was assessed using the number and duration of eye fixations. Our findings suggest that visual clutter has a negative influence on performance and alters the visual behavior during visual search in action video game scenes. Our results also suggest that expert action video game players might use different visual strategies to cope with clutter, leading however to no performance benefits.


Connectivity strength of the EEG functional network in schizophrenia and bipolar disorder.
The application of graph theory measures in the study of functional brain networks allows for the description of their general properties and their alterations in mental illness. Among these measures, connectivity strength (CS) estimates the degree of functional connectivity of the whole network. Previous studies in schizophrenia patients have reported higher baseline CS values and modulation deficits in EEG spectral properties during cognitive activity. The specificity of these alterations and their relationships with pharmacological treatments remain unknown. Therefore, in the present study, we assessed functional CS on EEG-based brain networks in 79 schizophrenia and 29 bipolar patients in addition to 63 healthy controls. The subjects performed a P300 task during the EEG recordings from which the pre-stimulus and the task-related modulation CS values were computed in the global and theta bands. These values were compared between the groups and between the patients who had and had not received different treatments. The global band pre-stimulus CS was significantly higher in the schizophrenia group compared with the bipolar and control groups. Theta band CS modulation was decreased in schizophrenia and bipolar patients. Treatment with antipsychotics, lithium, benzodiazepines, and anticonvulsants did not significantly alter these CS values. The first-episode and chronic schizophrenia patients did not show significant differences in CS values. Higher global band pre-stimulus CS values were associated with worse general cognition in schizophrenia patients. These data support increased connectivity in the whole-brain network that is specific to schizophrenia and suggest a general hyper-synchronized basal state that might hamper cognition in this syndrome.


The eruption of third molars in adults: a 10-year longitudinal study.
Among 829 participants of the Veterans Administration Dental Longitudinal Study examined over four triennial cycles, 97 persons (11.7%) had one or more unerupted third molars, for a total of 151 teeth, when the study was initiated. During the first 10 years of the study, 10 subjects had 14 unerupted third molars extracted, while in 10 persons 11 third molars erupted. Four of the latter persons each had one third molar removed subsequent to eruption. Three of the 11 erupted third molars were in functional occlusion. The position of 4 of the 11 "erupting" third molars appeared to remain unchanged relative to radiographic landmarks. Thus, their apparent clinical eruption may have been due to a reduction in the level of the gingival tissue in the area distal to the second molar tooth in association with periodontitis. The eruption of third molars in older adults appears to be a more frequent phenomenon than was previously recognized. Most erupting third molars had some associated pathologic condition, including caries and periodontitis.


Synthesis and cytotoxicity of acetyl-4H,9H-naphtho[2,3-b]thiophene-4,9-diones.
Several new acetyl-4H,9H-naphtho[2,3-b]thiophene-4,9-diones were synthesized and evaluated for in vitro cytotoxicity by NCI against seven cancer cell types. 2,7-Diacetyl naphtho[2,3-b]thiophene-4,9-dione (9) showed significant cytotoxicity against leukemia cells with log GI50 values of -7.61 against SR cells and -7.18 against MOLT-4 cells. 3-Acetyl-naphtho[2,3-b]thiophene-4,9-dione (6) also demonstrated potent cytotoxicity in the latter cell line with log GI50 < -8.


Assessment and treatment of psychotic speech in an autistic child.
The psychotic speech of autistic and other developmentally disabled children can be defined as words or phrases that are intelligible, but appear out of context. In the present investigation we conducted an analysis of the psychotic speech of a 9-year-old autistic boy. Three experiments were constructed to determine the functional significance of this child's psychotic speech and a method of intervention. The first study involved an analysis of the role of adult attention and task demands in the maintenance of psychotic speech. When task demands were increased, the frequency of psychotic speech increased. Varying adult attention had no effect on psychotic speech. We then performed a second analysis in which the consequence for psychotic speech was a 10-second time-out. Psychotic speech increased, suggesting that it may have been maintained through escape from task demands. Finally, the third experiment involved teaching an appropriate escape response ("Help me"). Psychotic speech was greatly reduced by this intervention. Thus, teaching an appropriate equivalent phrase proved to be a viable alternative to interventions using aversive consequences. The present study represents the first observation that psychotic speech may serve to remove children from unpleasant situations and also introduces a nonaversive intervention for this behavior.


The effect of a temporary nasoantral window on mucociliary clearance. An experimental study.
Although many studies have clearly shown that nasoantral windows do not help ciliary clearance of sinus disease, studies have not addressed the possible adverse effects of nasoantral windows. This study was performed to determine if an inferior antrostomy can actually improve ciliary clearance. Even with the advent of endoscopic sinus surgery we need to have a better understanding of inferior antrostomies.


Endoscopy-aided retrieval of broken drainage tube after lumbar spine surgery.
A ruptured drainage tube which remains in the incision is a rare surgical complication. The usual mode of retrieval is to detach the suture and explore the pre-existing incisional wound. However, spinal endoscopy provides an alternative method for successful removal, avoiding the enlargement of the surgical wound. A 53-year-old male patient underwent open lumbar spine surgery for lumbar spondylolisthesis between the 5th lumbar and 1st sacral vertebral bodies. Prior to closure, two negative pressure ball drainage tubes were inserted, one of which broke during removal,beneath the fascia. Use of spinal endoscopy enabled the complete removal of the broken drainage tube. Both the original incisional and endoscopic wounds healed well without any sign of infection. The use of spinal endoscopy to remove the broken drainage tube is an alternative to open the surgical wound and should be took into account.


Spinal Cord Injury With Tetraplegia in Young Persons After Diving Into Shallow Water: What Has Changed in the Past 10 to 15 Years?
Retrospective, monocentric, observational study in a tertiary health care center. To analyze prehospital and clinical findings, complications, neurological improvement and follow-up in a young person cohort with spinal cord injury (SCI) and tetraplegia according to the American Spinal Injury Association (ASIA) Impairment Scale (AIS) A to D after diving into shallow water. Included were all persons younger than 50 years with SCI after head-first diving into shallow water between June 2001 and June 2019. All persons with SCI were divided into complete tetraplegia (AIS A) and incomplete tetraplegia (AIS B, C, and D) to test differences. A total of 59 males (98.7%) and 1 female with a mean age of 27.7 years suffered an SCI. Alcohol use was documented in 25 cases (41.7%). At the time of admission, 33 people (55%) showed a complete tetraplegia (AIS A) and 27 showed an incomplete tetraplegia with 8 AIS B (13.3%), 15 AIS C (25%), and 4 AIS D (6.7%). At the time of discharge, people with initially complete tetraplegia showed a significant improvement from admission to discharge (P ≤ .004). Persons with incomplete tetraplegia were more likely to improve their neurological status compared with complete tetraplegia patients (P ≤ .001). Especially persons with complete tetraplegia suffered from typical SCI-related problems and complications. People with SCI and tetraplegia at the time admission show neurological improvement in 50% of the cases with an overall better outcome in persons with incomplete tetraplegia. The surgical treatment of SCI within 24 hours seems to be associated with a better neurological outcome and a lower level of tetraplegia. The incidence of SCI caused by diving into shallow water remains stable without a significant change, especially in high-risk groups. More education and prevention programs are necessary to avoid these injuries.