[Reliability of the abbreviated version of the Late Life Function and Disability Instrument--a meaningful and feasible tool to assess physical function and disability in the elderly].
The early assessment of prevalent disability and functional decline is of increasing importance for prevention issues and for the evaluation of change in clinical studies. The assessment of common daily tasks and resulting disability is essential for a complete impression of a patient's capabilities. There are plenty of instruments that cover basic and instrumental activities of daily living by assessing the person's dependence at a specific functional task. However, the early stages of everyday life limitations and functional decline are usually not being captured. In this article we have examined psychometric properties of the German adaptation of the abbreviated "late life function and disability instrument", a promising tool that is capable of capturing early aspects of functional decline and disability. Older Caucasian men and women (Median=82 years) with a wide range of functional status (Short Physical Performance Battery: Median=6, Min.=0, Max.=12) completed the abbreviated version of the LLFDI once (n=173). A subgroup was assessed twice to examine interrater (n=13) and intrarater (n=10) reliability. Cognitive and functional status was assessed at baseline. Internal consistency (Cronbach's ) was acceptable for the personal role and social role of the frequency items of the disability component (0.55 to 0.62) and very good for all other parts of the instrument (0.78 to 0.89) without a significant difference when compared for overall cognitive performance or executive function. Test-retest reliability was very good (0.81 to 0.96) and interrater reliability was acceptable to very good (0.62 to 0.96) with only acceptable results for the limitation items of the disability component. The total scores of the two components did not demonstrate significant floor or ceiling effects and the interview lasted on average 17 minutes (range 8 to 35 min). The German adaptation of the abbreviated version of the "Late Life Function and Disability Instrument" has minimal ceiling and floor effects, acceptable to good internal consistency and interrater reliability, and very good test-retest reliability. Hence, it constitutes an excellent alternative to common self-rating instruments for the assessment of functional capacities in everyday activities.


Enhancing the Interface Contact of Stacking Perovskite Solar Cells with Hexamethylenediammonium Diiodide-Modified PEDOT:PSS as an Electrode.
As an indispensable component of perovskite solar cells (PSCs), the commonly used Au and Ag electrodes still have some problems such as high cost and instability issues with regard to being corroded by iodide ions. In this paper, we report stacking perovskite solar cells (S-PSCs), which can avoid the use of precious metal electrodes and reduce the cost of devices and the requirements of equipment compared to conventional PSCs. The S-PSCs are composed of two semicells: a photoanode and a counter electrode (CE). For stacked devices, effective contact of the photoanode/CE interface is very important to the performance of the device. We used poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) as the electrode and modified it by hexamethylenediammonium diiodide (HDADI2) to improve its physical and electrical properties. The surface of the HDADI2-modified PEDOT:PSS becomes rough and achieves higher adhesion, which enables the photoanode and CE to be sufficiently connected. In addition, the energy-level structure of the HDADI2-modified PEDOT:PSS matches better with that of the adjacent functional layers. Therefore, the S-PSCs performance has been significantly improved. Under an illumination area of 1 cm2, the power-conversion efficiency (PCE) of the S-PSCs can reach 15.21%. Moreover, the S-PSCs can be disassembled and assembled flexibly and repeatedly disassembled 500 times with almost no change in the PCE. This has a positive impact on cell maintenance and modular production.


A framework for the analysis of polar anticancer drugs in wastewater: On-line extraction coupled to HILIC or reverse phase LC-MS/MS.
With the consumption of chemotherapy agents, residues of anticancer drugs may be increasingly found in hospital and municipal wastewaters. Quantification of these highly polar micropollutants remains challenging due to poor chromatographic retention on typical reversed phases. This study investigated different solid-phase extraction (SPE) materials for automated on-line preconcentration of complex matrices (hospital and municipal wastewaters) and various chromatographic column options. A hyper crosslinked hydroxylated polystyrene-divinylbenzene copolymer SPE sorbent coupled on-line with hydrophilic interaction liquid chromatography tandem mass spectrometry (HILIC-MS/MS) yielded suitable limits of detection (LOD: 1-2 ng L-1) for 5-fluorouracil (5-FU) and 2',2'-difluorodeoxyuridine (dFdU). Optimization of chromatographic conditions led to a single LC-MS/MS method for the analysis of other cytostatic drugs including cytarabine (CYT), gemcitabine (GEM), methotrexate (MTX), ifosfamide (IFO), cyclophosphamide (CYC) and capecitabine (CAP). The filter membrane for sample pre-treatment, HPLC mobile phase additives, and on-line SPE loading parameters were also investigated. The methods were validated in wastewater matrix with suitable determination coefficients (R2 range: 0.9982-0.9999), LODs (0.5-5 ng L-1), accuracy (78-111%), intraday precision (2.6-12%), and interday precision (2.1-13%). The occurrence of cytostatic drugs was examined in field-collected water samples from hospital effluents and municipal wastewater treatment plants (WWTP) in Canada. CAP (3.7-64 ng L-1), dFdU (6.1-300 ng L-1), and MTX (1.8-68 ng L-1) were frequently detected across both matrix types, while IFO was detected in hospital wastewater (23-140 ng L-1) but not in municipal WWTPs.


A modular, reversible sealing, and reusable microfluidic device for drug screening.
Preclinical tests for evaluating potential drug candidates using conventional protocols can be exhaustive and high-cost processes. Microfluidic technologies that can speed up this process and allow fast screening of drugs are promising alternatives. This work presents the design, concept, and operational conditions of a simple, modular, and reversible sealing microdevice useful for drug screening. This microdevice allows for the operation of 4 parallel simultaneous conditions and can also generate a diffusive concentration gradient in sextuplicates. We used laminated polydimethylsiloxane (PDMSLAM) and glass as building materials as proof of concept. The PDMSLAM parts can be reused since they can be easily sterilized. We cultured MCF-7 (Michigan Cancer Foundation-7) breast cancer cells. Cells were exposed to a doxorubicin diffusive concentration gradient for 3 h. They were monitored by automated microscopy, and after data processing, it was possible to determine cell viability as a function of doxorubicin concentration. The reversible sealing enabled the recovery of the tested cells and image acquisition. Therefore, this microdevice is a promising tool for drug screening that allows assessing the cellular behavior in dynamic conditions and the recovery of cells for afterward processing and imaging.


Trend of Conflict Rate in Iran from 2014 to 2020: An Ecological Study.
Detecting the correlation of conflict rate within provinces over time provides a better understanding for health policymakers in identifying potential causes. The purpose of this study was to assess the trend of conflict rate in 31 provinces of Iran using the growth mixture model (GMM). This ecologic study was conducted based on the data obtained from the Iranian Legal Medicine Organization (ILMO) by gender and provinces between March 21, 2014 and March 21, 2020. First, the 7-year cumulative incidence rates were described; second, the trend of conflict rate was modeled by a linear mixed-effects model according to gender and overall; finally, distinct classes of provinces with similar conflict trends in seven years were identified using the GMM. The significance level was considered less than 0.05. Among provinces, Ardebil and Sistan Baluchistan had the highest and the lowest 7-year conflict incidence rates (95% CI) with 66.6 (52.38, 84.67) and 20.79 (13.53, 31.95) per 100000, respectively. The results of the linear mixed-effects model showed that the annual rate of conflict in Iran decreased by 0.37% from 2014 to 2017 and then slightly increased by 0.07% after 2017. In addition, the GMM results indicated that the trends for Iranian provinces can be clustered into four distinct classes. Our study showed the increasing growth of conflict in the last years in most provinces of Iran. Necessary interventions are important to prevent the rising conflict rate due to the various effects of conflict on psychological, social, and health factors.


Specific cancer immunotherapy adjunctive to surgery for advanced gastric cancer: a report of six long term survivors.
In order to improve the results with non-curatively resected advanced gastric cancer, we have tried specific cancer immunotherapy adjunctive to surgery. In this article, six long term survivors (median 6.2 years) are reported.


Monocyte-mediated immune defense against murine Listeria monocytogenes infection.
Infection of mice with Listeria monocytogenes induces a robust innate inflammatory response that restricts bacterial growth in the liver and spleen prior to the development of protective T cell responses. Ly6C(hi) monocytes contribute to the innate immune response following L. monocytogenes infection and in their absence, mice rapidly succumb to infection. Emigration of Ly6C(hi) monocytes from the bone marrow into the circulation is the first step in their recruitment to sites of L. monocytogenes infection and is triggered by CCL2- and CCL7-mediated stimulation of CCR2 chemokine receptors on monocytes. CCL2 expression by mesenchymal stem cells in the bone marrow, in response to TLR stimulation, drives monocyte emigration from cellular compartments into vascular sinuses of the bone marrow. In addition to TLR ligands, type I interferon-mediated signals can also drive monocyte emigration from the bone marrow during L. monocytogenes infection. Once Ly6C(hi) monocytes enter the bloodstream, trafficking to sites of infection in the liver and spleen is CCR2 independent. In the liver, CD11b on the monocyte and ICAM-1 on the surface of endothelial cells target Ly6C(hi) monocytes to foci of L. monocytogenes infection. At the site of infection, Ly6C(hi) monocytes undergo MyD88-dependent differentiation into TNF and iNOS-producing dendritic cells (TipDCs) and express MHC class II, B7.1, and CD40 on their cell surface. How TipDCs mediate bacterial clearance during early L. monocytogenes infection remains an active area of investigation.


[Pituitary investigations in idiopathic haemochromatosis (author's transl)].
Twelve patients with idiopathic hemochromatosis have been studied. Hypogonadotropic hypogonadism with normal prolactin appears to be a most constant finding. Other hypophysal functions are preserved. A rapid exploration of such patients with LHRH-TRH and insulin tolerance test is proposed.


Diagnostic accuracy of diastolic pressure ratio using a pressure microcatheter for intracoronary physiological assessment.
Recently, instantaneous wave-free ratio (iFR) has emerged as an alternative to the fractional flow reserve (FFR) for intracoronary physiological assessment. Although all diastolic resting indices are reportedly identical to the iFR, limited data exist on diastolic pressure ratio (dPR) measured using a microcatheter (dPRmicro). This study aimed to evaluate the diagnostic accuracy of dPRmicro compared to FFR measured using a microcatheter (FFRmicro) in real-world practice for intracoronary physiological assessment. This was a single-center, retrospective, observational study. We identified 103 consecutive suspected angina pectoris patients (107 lesions) who underwent dPRmicro and FFRmicro measurement using the Navvus® catheter at Takasaki Heart Hospital from March 2019 to June 2019. A total of 103 lesions in 103 patients were finally included in the study. The mean FFRmicro and dPRmicro values were 0.80 and 0.88, respectively. With an FFRmicro ≤ 0.80, the dPRmicro showed a diagnostic accuracy of 79.6%, sensitivity of 74.6%, specificity of 87.5%, positive predictive value of 90.4%, and negative predictive value of 68.6%. The area under the receiver operating characteristic (ROC) curve was 0.894 (95% confidence interval, 0.833-0.956), and the optimal cut-off value for dPRmicro derived from the ROC analysis was 0.90. dPRmicro and FFRmicro values were discordant in 21/103 cases (20.4%). As a multivariable logistic regression analysis was performed, the male sex (vs. female) had a statistically significant association with a dPRmicro-FFRmicro discordance (OR 4.91; 95% CI, 1.04-23.0; P = 0.044). No other factors were found to be significantly associated with the discordance. In conclusion, dPRmicro measured using a microcatheter had good diagnostic accuracy and correlation with FFRmicro, hence, it can be useful for making revascularization decisions. However, re-studies in larger populations will be needed to better understand the properties of diastolic resting index measured using a microcatheter in clinical settings.


Plasma concentration of cytokine antagonists in patients with infection following liver resection.
The interactions between endogenous cytokine antagonists, surgical injury and perioperative infection are poorly understood. Perioperative plasma concentrations of soluble tumour necrosis factor receptor p55 (STNFR p55), interleukin (IL) 1 receptor antagonist (IL-1ra) and IL-6 were measured by immunoassay in 29 consecutive patients undergoing liver resection. Plasma concentrations of STNFR p55, IL-1ra and IL-6 showed a striking increase immediately after operation (day 0) and on postoperative day 1. STNFR p55 concentrations correlated significantly with operating time (P< 0.05); IL-1ra levels with operating time, blood loss and weight of resected liver (P < 0.05); and IL-6 concentrations with operating time and blood loss (P< 0.05). Eight of the 29 subjects developed postoperative infections, of whom four had positive cultures of enteric organisms. Plasma STNFR p55 and IL-6 concentrations were significantly higher in the infected group than in the uninfected group (P < 0.05). STNFR p55 and IL-6 concentrations on day 0 differentiated between the two groups with a diagnostic accuracy of 79 and 83 per cent respectively. These results suggest that STNFR p55, IL-1ra and IL-6 are produced during operation in response to surgical stress. This production of STNFR p55 and IL-6 may be enhanced further by intraoperative bacterial translocation. Measurement of plasma STNFR and IL-6 concentrations may predict the development of postoperative infectious complications.


Analgesic effect of dextromethorphan in neuropathic pain.
Dextromethorphan, a clinically available N-methyl-D-aspartic acid (NMDA) receptor antagonist, has an analgesic effect in patients with diabetic neuropathy. The aim of this study was to evaluate the analgesic and adverse effects of a single high dose of dextromethorphan on spontaneous pain in patients suffering long-term neuropathic pain of traumatic origin. Fifteen patients with post-traumatic neuropathic pain participated in this placebo-controlled, double-blind, randomized crossover study. On two separate occasions, the participants received 270 mg of dextromethorphan hydrobromide or placebo. Pain intensity, adverse effects and serum concentrations of dextromethorphan and metabolites were registered. Dextromethorphan had a statistically significant analgesic effect compared with placebo, but the effect varied markedly among the patients. Light-headedness was the most important adverse effect reported. Extensive metabolizers of dextromethorphan had an apparently better analgesic effect than poor metabolizers. This report indicates that a single high dose of dextromethorphan has an analgesic effect in patients with neuropathic pain of traumatic origin. The main metabolite dextrorphan seems to be important for the analgesic effect. At the relatively high dose studied, the clinical usefulness of dextromethorphan is limited to that portion of the patient population experiencing analgesia without an unacceptable level of adverse effects.


Expedited gene delivery for osteochondral defect repair in a rabbit knee model: A one-year investigation.
To evaluate a single-step, gene-based procedure for repairing osteochondral lesions. Osteochondral lesions were created in the patellar groove of skeletally mature rabbits. Autologous bone marrow aspirates were mixed with adenovirus vectors carrying cDNA encoding green fluorescent protein (Ad.GFP) or transforming growth factor-β1 (Ad.TGF-β1) and allowed to clot. The clotted marrow was press-fit into the defects. Animals receiving Ad.GFP were euthanized at 2 weeks and intra-articular expression of GFP examined by fluorescence microscopy. Animals receiving Ad.TGF-β1 were euthanized at 3 months and 12 months; repair was compared to empty defects using histology and immunohistochemistry. Complementary in vitro experiments assessed transgene expression and chondrogenesis in marrow clots and fibrin gels. In a subsequent pilot study, repair at 3 months using a fibrin gel to encapsulate Ad.TGF-β1 was evaluated. At 2 weeks, GFP expression was seen at variable levels within the cartilaginous lesion. At 3 months, there was no statistically significant improvement (p > 0.05) in healing of lesions receiving Ad.TGF-β1 and variability was high. At 12 months, there were still no significant difference (p > 0.05) between the empty defects and those receiving Ad.TGF-β1 in the overall, cartilage, and bone scores. Variability was still high. In vitro experiments suggested that variability reflected variable transduction efficiency and chondrogenic activity of the marrow clots; using fibrin gels instead of marrow may address this issue but more research is needed. This approach to improving the repair of osteochondral lesions needs further refinement to reduce variability and provide a more robust outcome.


Application of extracellular lipopeptide biosurfactant produced by endophytic Bacillus subtilis K1 isolated from aerial roots of banyan (Ficus benghalensis) in microbially enhanced oil recovery (MEOR).
Bacillus subtilis K1 isolated from aerial roots of banyan tree secreted mixture of surfactins, iturins and fengycins with high degree of heterogeneity. The extracellular extract consisting of mixture of these cyclic lipopeptides exhibited very good emulsification activity as well as excellent emulsion stability. The culture accumulated maximum surfactant up to 48 h of growth during batch fermentation in Luria broth. The emulsion of hexane, heptane and octane prepared using 48-h-old culture supernatant of B. subtilis K1 remained stable up to 2 days while emulsion of four stroke engine oil remained stable for more than a year. The critical micelle concentration of crude lipopeptide biosurfactant extracted by acid precipitation from 48-h-old fermentation broth of B. subtilis K1 was found to be 20.5 μg/mL. The biosurfactant activity was found to be stable at 100 °C for 2 h, over a pH range of 6-12 h and over an NaCl concentration up to 10 % (w/v). The application of biosurfactant on laboratory scale sand pack column saturated with four stroke engine oil resulted in ~43 % enhanced oil recovery, suggesting its suitability in microbially enhanced oil recovery.


[Histomorphology of the bone-cement contact. Experimental phenomenology of the osseous reconstruction processes. I].
In animal experiments the femurs of 25 rabbits were filled with bone cement. In cross sections of the diaphysis even after only one week, small gaps were found filled by lamellar formation of new bone. Osteoblast layers directly on the cement surface were also observed. After 4 to 6 weeks there was a circularly closed contact of inner lamellae and after 9 months a secondary medullary cavity was formed. At this time there was formed a trabcular contact with the implant, interrupted by medullary spaces full of cells. Sometimes woven-bone formations were found in the bone-cement interface.


Determination of ochratoxin A in liquorice products using HPLC based analytical methods. Part I: proficiency test of methods commonly used by the confectionary industry.
A European proficiency test series was accomplished on behalf of the CAOBISCO (Association of the Chocolate, Biscuit and Confectionery Industries of the EU) expert group on ochratoxin A to assess the performance of laboratories in measuring ochratoxin A in samples of various liquorice products. In addition, the impact of the extraction type (mainly with or without the use of halogenated solvents) was to be evaluated. Four different test samples (two liquorice powders and two liquorice pastes) were tested for sufficient homogeneity and distributed to 15 laboratories in 8 countries in Europe. The results were analysed using standard proficiency testing statistical procedures and laboratories were awarded z-scores on the basis of their reported results. The overall evaluation of the results shows a distinct variation between the participating laboratories. Based on a target standard deviation (σ-value) taken from the Horwitz equation, of the 14 laboratories that reported results, satisfactory results (z-score: |Z| ≤ 2.0) were obtained by 60% and 27% of the laboratories, respectively, for the two liquorice powders and by 93% and 53% of the laboratories, respectively, for the two liquorice pastes. Approximately equal numbers of laboratories used extraction types with and without the use of halogenated solvents. ANOVA testing of the results indicates there was no evident trend using different extraction solvents.


Genetic transformation of the white-rot fungus Dichomitus squalens using a new commercial protoplasting cocktail.
D. squalens, a white-rot fungus that efficiently degrades lignocellulose in nature, can be used in various biotechnological applications and has several strains with sequenced and annotated genomes. Here we present a method for the transformation of this basidiomycete fungus, using a recently introduced commercial ascomycete protoplasting enzyme cocktail, Protoplast F. In protoplasting of D. squalens mycelia, Protoplast F outperformed two other cocktails while releasing similar amounts of protoplasts to a third cocktail. The protoplasts released using Protoplast F had a regeneration rate of 12.5% (±6 SE). Using Protoplast F, the D. squalens monokaryon CBS464.89 was conferred with resistance to the antibiotics hygromycin and G418 via polyethylene glycol mediated protoplast transformation with resistance cassettes expressing the hygromycin phosphotransferase (hph) and neomycin phosphotransferase (nptII) genes, respectively. The hph gene was expressed in D. squalens using heterologous promoters from genes encoding β-tubulin or glyceraldehyde 3-phosphate dehydrogenase. A Southern blot confirmed integration of a resistance cassette into the D. squalens genome. An average of six transformants (±2 SE) were obtained when at least several million protoplasts were used (a transformation efficiency of 0.8 (±0.3 SE) transformants per μg DNA). Transformation of D. squalens demonstrates the suitability of the Protoplast F cocktail for basidiomycete transformation and furthermore can facilitate understanding of basidiomycete gene function and development of improved strains for biotechnological applications.


The impacts of polyethylene terephthalate microplastics (mPETs) on ecosystem functionality in marine sediment.
The effects of microplastics (MPs) on the ecological functioning in marine sediments is largely unknown. However, coastal marine sediments and their resident communities play critical roles in the transformation of organic matter and the cycling of nutrients that influence both local and global processes. To investigate how microplastics influence ecosystem functions associated with sediment biogeochemistry, large bivalves and microphytobenthos, we conducted a 31-day laboratory experiment. The experiment tested the role of micro-polyethylene terephthalate (mPETs) at five concentrations (0%, 1%, 3%, 6%, and 8% based on wet weight of top 1 cm sediment). Canonical principle of coordinate analysis (CAP) was applied to assess change on the ecosystem functionality with increasing concentrations of mPETs. Our results highlight that stress effects on ecosystem function are the product of the interaction between Macomona liliana, microphytobenthos and mPETs.


The utility of 18F-FDG PET/CT in solitary fibrous tumors of the pleura.
To demonstrate the utility of (18)F-FDG PET/CT in the differentiation of benign and malignant solitary fibrous tumors of the pleura (SFTP). A retrospective review was performed on the (18)F-FDG PET/CT data from 17 patients with histopathologically diagnosed benign or malignant SFTP. The size, side of SFTP, presence of necrosis, calcification, pleural effusion, hilar lymphadenopathy (LAP), density on CT images (Hounsfield unit-HU), and (18)F-FDG uptake (SUVmax) were recorded and compared in order to detect malignant SFTP. Statistical significance was set as p<0.05. The difference in size, presence of necrosis, and hilar LAP on CT images were statistically significant (p=0.004, p<0.001, p=0.015, respectively) in a comparison of benign and malignant SFTPs. The mean HU of benign SFTP was 46.16±5.52HU, and for malignant SFTP it was 35.03±4.61HU (p=0.003). The mean SUVmax was 3.02±1.02 for benign SFTP and 4.89±2.12 for malignant SFTP (p=0.021). A cut-off value of ≥7cm for size, ≤39.81HU for density, and ≥3.47 for SUVmax was obtained by ROC analysis for detecting malignant SFTP. (18)F-FDG PET/CT may have a limited role in diagnosing malignant SFTP in suspected patients.


Glucagon and GLP-1 inhibit food intake and increase c-fos expression in similar appetite regulating centres in the brainstem and amygdala.
Glucagon and glucagon-like peptide-1 (GLP-1) are evolutionarily related anorectic hormones. Glucagon also increases energy expenditure. The combination of glucagon and GLP-1 could cause weight loss through a simultaneous reduction in food intake and increased energy expenditure. However, the effect of combined administration of glucagon and GLP-1 on food intake and neuronal activation has not previously been studied. Furthermore, the effect of glucagon on neuronal activation in appetite regulating centres has not been assessed. Characterisation of the effects of glucagon when administered singly and in combination with GLP-1 on neuronal activation will be important for determining the mechanism of action of related potential antiobesity therapies. To investigate the effects of peripherally administered GLP-1 and glucagon on food intake, neuronal activation and blood glucose in mice when administered individually and in combination. Food intake, blood glucose and c-fos expression in the hypothalamus, amygdala and brainstem were measured in response to GLP-1 and glucagon, alone and in combination. Peripherally administered GLP-1 and glucagon decreased food intake and increased c-fos expression in the brainstem and amygdala. Doses of GLP-1 and glucagon that individually did not significantly affect feeding, in combination were anorectic and stimulated neuronal activation in the area postrema (AP) and central nucleus of the amygdala. Combined administration of GLP-1 and glucagon prevented the acute hyperglycemic effect of glucagon alone. Anorectic doses of glucagon and GLP-1 induced similar patterns of c-fos expression. Combined administration of low dose GLP-1 and glucagon inhibited food intake and induced c-fos expression in the AP and amygdala. The combination of both hormones may offer the opportunity to utilise the beneficial effects of reduced food intake and increased energy expenditure, and may therefore be a potential treatment for obesity.


Ultrastructural observations of the abducens nucleus of the cat after injection of horseradish peroxydase into the lateral rectus muscle.
The ultrastructure of cat abducens motoneurones is described, after labelling by retrograde axonal transport of horseradish peroxydase. The motoneurones were densely covered with synaptic boutons of both the "terminal" and "en passant" types and axo-axonal synapses were observed on the initial segment of motoneurone axons. Within the population of axons forming the VIth nerve, multiple branching was encountered and axons closely apposed at the nodes of Ranvier were seen.


The description and initial evaluation of an intensive live-in program in risk reduction.
Drawing upon the epidemiological, clinical, and behavioral studies suggesting the importance and feasibility of multidisciplinary efforts to reduce levels of risk variables for cardiovascular and other chronic diseases, a short-term live-in intervention program was developed. The program includes efforts to reduce smoking, weight, blood lipids, blood pressure, and stress through improving habits of exercise, nutrition, weight management, and stress control delivered to individuals with varying levels of health risk based on measurable biochemical and physiological variables and medical history. Major changes occurred during the 24-day program in 459 individuals enrolled in the program: 68% of smokers ceased, average cholesterol fell from 240 to 200 mg%, ideal body weight fell from 134 to 129% (82 to 79 kg), systolic blood pressure (BP) fell from 131 to 119 mm Hg, diastolic BP fell from 81 to 73 mm Hg. and reported feelings of general well-being increased. Greater changes were observed in the high-risk groups. Follow-up results at 1 year (48% of patients reporting) for those defined as high risk were a net decrease of 22 mg% in cholesterol, 7 mm Hg in systolic BP, 6 mm Hg in diastolic BP, and 6.8% of ideal body weight; 45% of those smoking at admission were still not smoking at 1 year (32% reporting). For those at lower risk there was a general return of risk levels toward baseline values.


Modulation of visceral nociceptive pathways.
Increased sensitivity of visceral nociceptive pathways contributes to symptoms in an array of clinical gastrointestinal conditions, however, the search for a consistently effective pharmacological agent to treat these conditions remain elusive. Modulation of visceral nociceptive pathways can occur at peripheral, spinal and supra-spinal sites and a dizzying array of potential drug targets exists. Till date, only tricyclic anti-depressants (TCAs) such as amitriptyline and, more recently, selective serotonin reuptake inhibitors (SSRIs) such as citalopram have demonstrated convincing visceral anti-nociceptive properties and clinical benefit in a limited population of patients with visceral hypersensitivity. Unfortunately, there is an incomplete understanding of the receptors and/or primary site of action at which these compounds exert their effects and significant side effects are often encountered. There is a continuing and concerted effort underway to develop target-specific visceral analgesic/anti-hyperalgesic compounds and the aim of this article is to provide a concise update on the most recent advances in this area.


Expression of mRNA for the t-complex polypeptide-1, a subunit of chaperonin CCT, is upregulated in association with increased cold hardiness in Delia antiqua.
Summer-diapause and winter-diapause pupae of the onion maggot, Delia antiqua (Diptera: Anthomyiidae), were significantly more cold hardy than nondiapause, prediapause, and postdiapause pupae. Moreover, cold acclimation of nondiapause pupae conferred strong cold hardiness comparable with that of diapause pupae. Differential display analysis revealed that the expression of a gene encoding TCP-1 (the t-complex polypeptide-1), a subunit of chaperonin CCT, in D antiqua (DaTCP-1) is upregulated in the pupae that express enhanced cold hardiness. Quantitative real-time polymerase chain reaction analyses showed that the levels of DaTCP-1 messenger RNA in pupal tissues, brain, and midgut in particular, are highly correlated with the cold hardiness of the pupae. These findings suggest that the upregulation of DaTCP-1 expression is related to enhanced cold hardiness in D antiqua. The upregulation of CCT in response to low temperature in an organism other than the yeast is newly reported.


Highly sensitive and selective method to detect dopamine in the presence of ascorbic acid by a new polymeric composite film.
A new approach was applied to modify gold electrode with a unique polymer composite for selectively detecting dopamine (DA), a neurotransmitter, in the presence of an electroactive species of ascorbic acid (AA). After self-assembly of 11-mercaptoundecanoic acid (MUA) monolayer on gold surface, polyethylene glycol (PEG) was used to perform electrochemical esterification with MUA. In general, AA is the main interference of DA detection in a biological system. The resulting composite layer showed high sensitivity to detect DA but selectively blocked the interference from AA. Furthermore, for the first time, an interesting mechanism was demonstrated from our experimental results, namely, that the catalytic effect of AA on DA is limited by DA concentration when AA/DA>1. The modified electrode showed good reproducibility (+/-2% relative standard deviation), a low detection limit (10 nM), a fast response time (<2s), high sensitivity (86 nA/microM), a wide dynamic range of detection (20 microM), and great selectivity (without AA interference). The discovery is very promising for applications of detection of DA in a physiological environment where a high concentration of AA always exists.


Serglycin: the master of the mast cell.
Serglycin is a proteoglycan composed of a relatively small (~17 kDa) core protein to which sulfated glycosaminoglycans of either heparin, heparan sulfate or chondroitin sulfate types are attached. Serglycin is expressed in many cell types, including in particular cells of hematopoietic origin. To study the function of serglycin, we have used a serglycin knockout mouse strain. A striking finding was that the mast cell population was severely affected by the absence of serglycin, as evidenced by distorted granule morphology and defective staining with cationic dyes. Moreover, the absence of serglycin caused a dramatic effect on the ability of mast cells to store a number of granule compounds, including several mast cell-specific proteases as well as biogenic amines. Hence, serglycin has a major function in maintaining mast cell secretory granule homeostasis.


Synthesis, Characterization and Catalytic Application of Pyridine-Bridged N-Heterocyclic Carbene-Ruthenium Complexes in the Hydrogenation of Carbonates.
A series of bulky pyridine-bridged NHC-Ru complexes have been rationally designed and synthesized; these exhibited very high catalytic activity in the hydrogenation of cyclic and linear carbonates under mild reaction conditions. In the presence of catalytic amounts of a weak base, a broad range of substrates with different ring size and steric bulk were well tolerated, providing methanol and the corresponding diols in excellent yields with a catalyst loading as low as 0.5 mol %.


Mining Xanthomonas and Streptomyces genomes for new pectinase-encoding sequences and their heterologous expression in Escherichia coli.
Microbial genome sequencing has left a legacy of annotated yet uncharacterized genes or open reading frames, activities that may have useful applications in health and/or the environment. We are interested in the discovery and characterization of potentially new pectinolytic activities for the enzymatic retting of natural bast fibers such as hemp and flax. A highlight in this study is the discovery of a cold-active pectate lyase among five pectate-lyase-encoding sequences and two polygalacturonase-encoding sequences that we have cloned from the genomes of Xanthomonas campestris pv. campestris and Streptomyces coelicolor A3(2). Heterologous expression of these sequences as active pectate lyases and polygalacturonases required their subcloning in Escherichia coli Rosetta cells. The most active recombinant pectate lyase (XcPL NP_638163), a cold-active pectate lyase (XcPL NP_636037), and a polygalacturonase (XcPG NP_638805) were purified to near homogeneity and their kinetic parameters were determined. A significant amount of pectin degradation products was shown to be released by the two pectate lyases but not the polygalacturonase when hemp fiber pectin was used as substrate. Results of this study showed that genome data mining, besides an economical approach to new gene acquisition, may uncover new findings such as the discovery of a cold-active pectate-lyase-encoding sequence from X. campestris, a mesophilic microorganism.


Reconstruction of an enterocutaneous fistula using a superior gluteal artery perforator flap.
Enterocutaneous fistula is an uncommon complication of surgery for colorectal cancer. However, once a fistula has developed, treatment is complicated by previous treatments. Here, we describe an enterocutaneous fistula that developed after multiple treatments for rectal cancer in a 62-year-old woman. The woman had previously undergone several colorectal surgeries, radiation therapy and five courses of chemotherapy. Four years after the final surgery, an enterocutaneous fistula developed between the small intestine and the sacral skin. The fistula was resected, and the resulting defect was successfully reconstructed with a superior gluteal artery perforator flap.


Hedgehog regulates smoothened activity by inducing a conformational switch.
Hedgehog (HH) morphogen is essential for metazoan development. The seven-transmembrane protein smoothened (SMO) transduces the HH signal across the plasma membrane, but how SMO is activated remains poorly understood. In Drosophila melanogaster, HH induces phosphorylation at multiple Ser/Thr residues in the SMO carboxy-terminal cytoplasmic tail, leading to its cell surface accumulation and activation. Here we provide evidence that phosphorylation activates SMO by inducing a conformational switch. This occurs by antagonizing multiple Arg clusters in the SMO cytoplasmic tail. The Arg clusters inhibit SMO by blocking its cell surface expression and keeping it in an inactive conformation that is maintained by intramolecular electrostatic interactions. HH-induced phosphorylation disrupts the interaction, and induces a conformational switch and dimerization of SMO cytoplasmic tails, which is essential for pathway activation. Increasing the number of mutations in the Arg clusters progressively activates SMO. Hence, by employing multiple Arg clusters as inhibitory elements counteracted by differential phosphorylation, SMO acts as a rheostat to translate graded HH signals into distinct responses.


Fine-needle aspiration cytology diagnosis of non-Hodgkins lymphoma in a resource-challenged environment.
To establish the role of fine-needle aspiration cytology (FNAC) as a diagnostic tool for non-Hodgkins lymphoma in a resource challenged environment. This study was conducted on patients with lymphadenopathy, attending various clinics over a period of 18 months. FNAC of the enlarged lymph nodes was performed and biopsy, special stains and immunohistochemical staining was done in selected cases. Out of the total 275 cases, 42 cases (16%) were primary lymphoproliferative disorders. Non-Hodgkin lymphoma comprised of 32 cases (76.2% of all lymphomas), Hodgkin lymphoma-10 cases and the rest were metastatic carcinoma. The diagnostic accuracy for non-Hodgkin Lymphoma was 93.3%, sensitivity 95.4%, and specificity 87.5%. FNAC is a rapid, safe, easy, and nonexpensive diagnostic technique which can be used for early diagnosis of non-Hodgkins lymphoma.


Increasing TRPV4 expression restores flow-induced dilation impaired in mesenteric arteries with aging.
The flow-stimulated intracellular Ca(2+) concentration ([Ca(2+)]i) rise in endothelial cells is an important early event leading to flow-induced blood vessel dilation. Transient receptor potential vanilloid subtype 4 (TRPV4), a Ca(2+)-permeable cation channel, facilitates the flow-stimulated [Ca(2+)]i rise. To determine whether TRPV4 is involved in age-related flow-induced blood vessel dilation impairment, we measured blood vessel diameter and nitric oxide (NO) levels and performed Ca(2+) imaging, immunoblotting, and immunostaining assays in rats. We found that the flow-induced and TRPV4 activator 4α-PDD-induced dilation of mesenteric arteries from aged rats were significantly decreased compared with those from young rats. The flow- or 4α-PDD-induced [Ca(2+)]i rise was also markedly reduced in primary cultured mesenteric artery endothelial cells (MAECs) from aged rats. Immunoblotting and immunostaining results showed an age-related decrease of TRPV4 expression levels in MAECs. Additionally, the 4α-PDD-induced NO production was significantly reduced in aged MAECs. Compared with lentiviral GFP-treated aged rats, lentiviral vector delivery of TRPV4 increased TRPV4 expression level in aged MAECs and restored the flow- and 4α-PDD-induced vessel dilation in aged mesenteric arteries. We concluded that impaired TRPV4-mediated Ca(2+) signaling causes endothelial dysfunction and that TRPV4 is a potential target for clinical treatment of age-related vascular system diseases.


A Descriptive Study of Home Modifications for People with Dementia and Barriers to Implementation.
This study describes home environmental features, safety issues, and health-related modifications in a community dwelling sample of 82 elderly people with dementia. Main barriers to the accessibility of the homes were steps, both inside and outside the house. The majority of the caregivers had made home modifications, which pertained mainly to physical limitations. Home modifications to support cognitive deficits were made to a lesser extent. The main barrier to the implementation of home modifications to accommodate the care recipient's memory loss was skepticism about their usefulness. Regarding the removal of physical barriers, financial constraints were most frequently mentioned.


Transcriptomic analysis of hepatocytes reveals the association between ubiquitin-specific peptidase 1 and yes-associated protein 1 during liver regeneration.
The liver has an excellent ability to regenerate, and disrupted liver regeneration after various injuries leads to an unfavorable prognosis for patients. In this study, we sought to identify novel therapeutic hallmarks that are associated with yes-associated protein 1 (YAP1)-mediated hepatocyte proliferation during the process of liver regeneration. Partial hepatectomy was conducted to induce liver regeneration in rats. Primary hepatocytes were isolated and cultured. Hepatocyte proliferation was assessed using immunohistochemistry staining, and expression of YAP1 was detected. RNA sequencing and bioinformatics analysis were used to search for potential regulators of YAP1. The association between ubiquitin-specific peptidase 1 (USP1) and YAP1 was validated using in vivo and in vitro experiments. YAP1 was significantly elevated in regenerative hepatocytes, especially in the nucleus. Knockdown of YAP1 using small interfering RNA or pharmacological inhibition using verteporfin significantly attenuated the proliferation of hepatocytes. The bioinformatics analysis results revealed that USP1 was associated with YAP1-mediated hepatocyte proliferation during liver regeneration. ML-323, a specific inhibitor of USP1-USP1 associated factor 1 (UAF1), significantly decreased the expression of YAP1, Cyclin D1, and proliferating cell nuclear antigen, while these decreased expressions could be rescued by YAP1 overexpression. Furthermore, ML-323 treatment significantly inhibited liver regeneration following partial hepatectomy. In conclusion, we identified USP1 as a novel biomarker that is associated with YAP1-mediated hepatocyte proliferation in liver regeneration. Pharmacological inhibition of USP1 by ML-323 substantially impairs hepatocyte proliferation during liver regeneration.


Hepatocellular carcinoma in the U.S.A., etiologic considerations. Localization of hepatitis B antigens.
The serologic and tissue markers of hepatitis B virus (HBV) were studied in 50 patients in whom hepatocellular carcinoma (HCC) was confirmed at autopsy. Serologic and tissue markers included serum hepatitis B surface antigen (HBsAg), tissue HBsAg, tissue hepatitis core antigen (HBcAg), and serum antibody to HBcAg (anti-HBc). Twenty-two patients had HCC arising in alcoholic cirrhosis; 2 of the 22 (9.1%) had one or more of the HBV tissue and serologic markers. This infection rate is similar to the rate of 7.9% observed in 63 control alcoholic cirrhotic patients without HCC. In contrast, 15 of 20 (75.0%) patients with HCC in nonalcoholic chronic active liver disease showed evidence of active HBV infection. One of 8 patients with HCC in normal liver had serum HBV markers. This result indicates that there is an extremely high prevalence of HBV infection among HCC patients with nonalcoholic chronic liver disease in the U.S.A. The prevalence of HBV infection in these patients is as high as that observed in Asia and Africa. Thus, it can be concluded that the lower prevalence rate of active HBV infection in HCC patients in the U.S.A. is the result of statistical dilution of HCC-B-viral disease by the large numbers of the alcoholic cirrhotic patients with HCC, and that if chronic active hepatitis type B were as common in the United States as it is in Africa and Asia, the frequency of occurrence of HCC might also be as high.


Shaping modern human skull through epigenetic, transcriptional and post-transcriptional regulation of the RUNX2 master bone gene.
RUNX2 encodes the master bone transcription factor driving skeletal development in vertebrates, and playing a specific role in craniofacial and skull morphogenesis. The anatomically modern human (AMH) features sequence changes in the RUNX2 locus compared with archaic hominins' species. We aimed to understand how these changes may have contributed to human skull globularization occurred in recent evolution. We compared in silico AMH and archaic hominins' genomes, and used mesenchymal stromal cells isolated from skull sutures of craniosynostosis patients for in vitro functional assays. We detected 459 and 470 nucleotide changes in noncoding regions of the AMH RUNX2 locus, compared with the Neandertal and Denisovan genomes, respectively. Three nucleotide changes in the proximal promoter were predicted to alter the binding of the zinc finger protein Znf263 and long-distance interactions with other cis-regulatory regions. By surface plasmon resonance, we selected nucleotide substitutions in the 3'UTRs able to affect miRNA binding affinity. Specifically, miR-3150a-3p and miR-6785-5p expression inversely correlated with RUNX2 expression during in vitro osteogenic differentiation. The expression of two long non-coding RNAs, AL096865.1 and RUNX2-AS1, within the same locus, was modulated during in vitro osteogenic differentiation and correlated with the expression of specific RUNX2 isoforms. Our data suggest that RUNX2 may have undergone adaptive phenotypic evolution caused by epigenetic and post-transcriptional regulatory mechanisms, which may explain the delayed suture fusion leading to the present-day globular skull shape.


Effects of nitrate on the saxitoxins biosynthesis revealed by sxt genes in the toxic dinoflagellate Alexandrium pacificum (group IV).
The dinoflagellate Alexandrium pacificum (group IV) is of particular interest because of its involvement in harmful algal blooms and production of saxitoxin (STX), which causes paralytic shellfish poisoning. The toxicity from STX and its analogues (STXs) is suspected to be affected by nitrogen (N) availability. However, the toxicity-associated behavior and STX-biosynthesis gene responses of the toxic A. pacificum under N fluctuations have not been sufficiently investigated. In the present study, we identified the sxtI gene involved in sxt biosynthesis pathway and evaluated the effects of nitrate (NO3-) on STXs production and the expression of four sxt core genes (sxtA4, sxtG, sxtB, and sxtI). Quantification of total STXs levels in the cultures under different NO3- regimes showed that NO3- concentration influenced STXs production. In addition, the proportion and concentration of STXs varied depending on the NO3- concentration. Core sxt transcript abundance was also influenced by available NO3- in a time-dependent manner. Expressional levels and patterns of sxtI were correlated with those of sxtA and sxtB. The relationship between the toxins and sxt responses in A. pacificum under various NO3- regimes suggests the direct involvement of N in the STXs biosynthesis pathway. Understanding this link would provide a tool to understand the toxin dynamics of dinoflagellates following N shifts in marine environments.


Phylogeny of anion exchangers: could trout AE1 conductive properties be shared by other members of the gene family?
A phylogenetic tree of anion exchangers (AE) was performed in order to better understand relationships between the different known AE and how they arose. Indeed, the different known AE1 from mammals or fish do not exhibit the same transport features: all studied anion exchangers 1 (AE1) catalyse an electroneutral Cl-/HCO3- exchange through the plasma membrane; however, trout AE1 (tAE1) is able to spontaneously form an anion conductive pathway permeable to some inorganic cations (Na+ and K+) as well as to organic osmolytes such as taurine. Therefore, it has been proposed that this major erythrocyte membrane protein could play a key role for the cell volume regulation of trout red cells. By analogy, it was envisioned that other fish anion exchangers could play a similar role in osmolyte loss induced by erythrocyte swelling. We have cloned AE1 from Raja erinacea and Danio rerio and studied their properties after expression in Xenopus laevis oocytes. In this study, we show that none of them is able to induce any conductive pathway or taurine permeability in Xenopus oocytes. Our phylogenetic analyses show that, first, all present AE1 genes have a common ancestor distinct from that of AE2 and AE3 and second, tAE1 is a true AE1 ortholog. The question of whether tAE1 conductive properties are a derived character in the trout lineage within Euteleostei or whether other AE1 members can share these properties is then discussed.


The Nord Stream pipeline gas leaks released approximately 220,000 tonnes of methane into the atmosphere.
Sudden mega natural gas leaks of two Nord Stream pipelines in the Baltic Sea (Denmark) occurred from late September to early October 2022, releasing large amounts of methane into the atmosphere. We inferred the methane emissions of this event based on surface in situ observations using two inversion methods and two meteorological reanalysis datasets, supplemented with satellite-based observations. We conclude that approximately 220 ± 30 Gg of methane was released from September 26 to October 1, 2022.


cis-B7:CD28 interactions at invaginated synaptic membranes provide CD28 co-stimulation and promote CD8+ T cell function and anti-tumor immunity.
B7 ligands (CD80 and CD86), expressed by professional antigen-presenting cells (APCs), activate the main co-stimulatory receptor CD28 on T cells in trans. However, in peripheral tissues, APCs expressing B7 ligands are relatively scarce. This raises the questions of whether and how CD28 co-stimulation occurs in peripheral tissues. Here, we report that CD8+ T cells displayed B7 ligands that interacted with CD28 in cis at membrane invaginations of the immunological synapse as a result of membrane remodeling driven by phosphoinositide-3-kinase (PI3K) and sorting-nexin-9 (SNX9). cis-B7:CD28 interactions triggered CD28 signaling through protein kinase C theta (PKCθ) and promoted CD8+ T cell survival, migration, and cytokine production. In mouse tumor models, loss of T cell-intrinsic cis-B7:CD28 interactions decreased intratumoral T cells and accelerated tumor growth. Thus, B7 ligands on CD8+ T cells can evoke cell-autonomous CD28 co-stimulation in cis in peripheral tissues, suggesting cis-signaling as a general mechanism for boosting T cell functionality.


Significance of the micropores electro-sorption resistance in capacitive deionization systems.
Capacitive Deionization (CDI) is an emerging technology representing a potential alternative to the common, energy-intensive desalination methods for low salinity water streams. In CDI an electrical field is applied to separate ionic species from aqueous solutions and electro-adsorb them into a highly porous material. CDI is a complex multi-scale system which requires robust mathematical models to closely describe its performance. Here, a dynamic two-dimensional model is developed coupling the diffusion and advection of the species in the bulk solution with their diffusion and electro-sorption in the porous electrodes. In this model, the adsorption/desorption resistance between the micropores and macropores along with variable non-electrostatic attractive forces in the micropores are also incorporated. The proposed theory is validated against experiments using a circular CDI cell operating under various conditions, where different transport mechanisms are limiting the total ion removal process. Performance of the CDI systems is also evaluated using inclusive figures of merit. The obtained results accentuate the significant effect of the rate-limited transfer of the ionic species from the macropores into the micropores, especially in systems subject to severe ion starvation, where neglecting this electro-sorption resistance leads to up to 50% and 210% overestimation of the energy efficiency and overall desalination performance, respectively. Furthermore, although the commonly used transport theory describing CDI fails to capture the dynamics of the systems at low initial concentration and high adsorption capacity by assuming fast electro-sorption without any resistance, the presented theory closely models the transport mechanisms in such systems. Moreover, we experimentally and numerically demonstrate a trade-off between the energetic and desalination performance in systems with low and high mass Péclet number.


Glucocorticoid, androgen, and retinoic acid regulation of glutathione S-transferase gene expression in hamster smooth muscle tumor cells.
A mu class glutathione S-transferase gene (hGSTYBX) is expressed in the DDT1MF-2 hamster smooth muscle tumor cell line. This gene is glucocorticoid responsive, and near maximal induction was found to occur within 24 h. The induced mRNA was very stable with a half-life of more than 48 h. Serum had no effect on either constitutive or glucocorticoid induced hGSTYBX expression. Although dibutyryl cAMP, phenobarbital, and 12-O-tetradecanoylphorbol-13-acetate did not alter hGSTYBX expression, testosterone and retinoic acid were each able to increase hGSTYBX expression in a concentration dependent manner. These results demonstrate a unique pattern of responsiveness of the hamster gene compared to the glutathione S-transferase genes of other species.


Structural properties of the glial fibrillary acidic protein. Evidence for intermolecular disulfide bonds.
Glial fibrillary acidic protein was purified from 4 M urea extracts of bovine brain by DEAE Bio-Gel A chromatography, 30% ammonium sulfate precipitation and hydroxylapatite chromatography. Subunits of about 54 000 daltons are present in solution as polydisperse distributions of polymers largely constrained by the presence of interchain disulfide linkages. Circular dichroism measurements indicate a native conformation containing some alpha-helical structure. The relevance of these findings to the cytoskeletal function of intermediate (80-100 A) filaments is discussed.


Radiographic detection of accessory/lateral canals: use of RadioVisioGraphy and Hypaque.
The diagnostic accuracy of RadioVisioGraphy (RVG-S) and conventional film images with and without the use of a radiopaque contrast media were compared for the detection of accessory/lateral canals. Forty-nine root canals, 13 with verified lateral canals, were endodontically prepared and imaged using both conventional intraoral dental film (E-speed) and three modes of RVG-S digital radiography (normal, inverse, zoom) before and after introduction of Hypaque-M, 90%. Images were presented to seven viewers; however, three demonstrated only fair intrarater reliability and were excluded. Overall sensitivity was low. RVG images were slightly more sensitive than conventional film images. Specificities, positive predictive values, and negative predictive values were low and showed no differences between modalities. It was concluded that the diagnostic accuracy of both film and RVG-S digital radiographic images for the detection of accessory/lateral canals, either alone or in combination with radiopaque contrast material, is low.


Inherited Selenocysteine Transfer RNA Mutation: Clinical and Hormonal Evaluation of 2 Patients.
Iodothyronine deiodinases are selenoproteins with the amino acid selenocysteine (Sec) introduced into the position of a TGA stop codon by a complex machinery involving tRNA[Ser]Sec when a cis-acting Sec-insertion sequence element is present in the 3' end of the mRNA. Recently, a variant in the TRU-TCA1-1 gene encoding for tRNA[Ser]Sec was reported, which resulted in reduced expression of stress-related selenoproteins. The proband presented with multisystem symptoms, euthyroid hyperthyroxinemia, and selenium deficiency. Here, we describe 2 new members of a family harboring the same tRNA[Ser]Sec variant. A 13-year-old patient was seen for Hashimoto's disease with high FT3 (4.6 pg/mL, normal range 2-4.2 pg/mL) and normal FT4 and TSH concentrations. He had no clinical complaints. During a 6-year clinical and hormonal follow-up, the index patient was not treated, FT3 decreased, FT4 increased, and serum TSH stayed in the normal range resulting in a euthyroid hyperthyroxinemia. Reverse T3 concentration was significantly increased at the last visit (19 years and 4 months). At the last evaluation, the total selenium level was low (91 μg/L, normal range 95-125). DNA sequencing identified a germinal homozygous variant (C65G) in the TRU-TCA1-1 gene. During follow-up, no additional clinical symptom was observed in the absence of any treatment. The same germinal tRNA[Ser]Sec variant was identified at heterozygous state in his father, who had normal thyroid function tests except a moderately increased reverse T3 concentration, with increased total selenium (143 μg/L) level. In both patients, the expression of stress-related selenoprotein GPX3 was in the low-normal range (168 and 180 IU/L, respectively, normal range: 150-558 IU/L). We did not find any significant biological abnormalities evocative of other selenoprotein deficiencies. We report on 2 members of a family with a variant in the TRU-TCA1-1 gene encoding for tRNA[Ser]Sec. Our study suggests that this tRNA[Ser]Sec variant is not exclusively causative of disruption in selenoprotein synthesis.


Buprenorphine treatment episode duration, dosage, and concurrent prescribing of benzodiazepines and opioid analgesics: The effects of Medicaid prior authorization policies.
Buprenorphine is an effective medication for the treatment of opioid use disorder (OUD), but the association between prior authorization policies and quality of care for individuals receiving buprenorphine treatment is not well-understood. Using 2006-2013 Medicaid Analytic eXtract (MAX) data from 34 states and the District of Columbia, we identified 294,031 episodes of buprenorphine treatment for OUD among individuals aged 14-64 years. We estimated generalized difference-in-differences models to examine the association between buprenorphine prior authorization policies and changes in buprenorphine treatment quality along four dimensions: (1) duration of at least 180 days, (2) dosage of at least 8 milligrams, and concurrent prescribing of (3) opioid analgesics and (4) benzodiazepines. Buprenorphine prior authorization policies were associated with an 11-percentage point reduction (p < 0.01) in the likelihood of episodes with a duration of at least 180 days in the first four years after policy implementation. The policy was not associated with changes in effective dosage or concurrent prescribing of opioid analgesics or benzodiazepines. Buprenorphine prior authorization policies were associated with a sizeable and significant reduction in episodes of at least 180 days duration, underscoring the importance of identifying and removing barriers to effective and appropriate OUD care.


Low ATM protein expression in malignant tumor as well as cancer-associated stroma are independent prognostic factors in a retrospective study of early-stage hormone-negative breast cancer.
The serine/threonine protein kinase ataxia telangiectasia mutated (ATM) is critical in maintaining genomic integrity. Upon DNA double-strand breaks, ATM phosphorylates key downstream proteins including p53 and BRCA1/2, thereby orchestrating complex signaling pathways involved in cell cycle arrest, DNA repair, senescence and apoptosis. Although sporadic mutation of ATM occurs rarely in breast cancer, the status of its protein expression and its clinical significance in breast cancer remain not well established. Our study was designed to investigate the influence of ATM protein in both tumor and cancer-associated stroma on clinical outcome in hormone-positive (HPBC) and hormone-negative (HNBC) early-stage breast cancer (EBC). Tissue microarrays (TMAs), containing formalin-fixed, paraffin-embedded resected tumors from two cohorts of patients (HPBC cohort: n=130; HNBC cohort: n=168) diagnosed at the Tom Baker Cancer Centre, Calgary, Canada, were analyzed for ATM protein expression using fluorescence immunohistochemistry (IHC) and automated quantitative analysis (AQUA). ATM expression levels were measured within the tumor as a whole (tATM) as indicated by pan-cytokeratin expression, tumor nuclear compartment (nATM) as indicated by both DAPI and pan-cytokeratin-positive results, and cancer-associated stroma (csATM) as indicated by vimentin-positive and pan-cytokeratin-negative results. ATM expression levels within these compartments were correlated with clinical outcome. While tATM and nATM were significantly lower in tumors compared to normal breast epithelial tissues, csATM was significantly higher than the corresponding normal tissue compartment. In addition, the median expression level of both tATM and nATM were two- to threefold lower (P<0.001) in HNBC than in HPBC. In both HNBC and HPBC cohorts, patients with low tATM, nATM and csATM tumors had significantly poorer survival outcomes than those with a high tATM, nATM and csATM, but this effect was more pronounced in HNBC. A multivariate analysis demonstrates that these biomarkers predict survival independent of tumor size and lymph node status, but only in the HNBC cohort (P<0.001). Low ATM protein expression in both malignant tumor and stromal compartments likely contributes to the aggressive nature of breast cancer and is an independent prognostic factor associated with worse survival in HNBC patients.


A biomechanical approach to the analysis of methods and procedures of bariatric surgery.
Bariatric surgery includes a variety of procedures that are performed on obese people and aim at decreasing the intake of food and calories. This goal is usually pursued by reducing stomach capacity and/or absorbing capability. Adjustable gastric banding is the most common and successful operation. In general, bariatric surgical procedures are effective, but are often associated with major complications. Surgical procedure and post-surgical conformation of the stomach are usually defined on clinical and surgical basis only. Instead, the optimal configuration should be identified by analyzing the mechanical functionality of the stomach and the surrounding structures, and the relationship between food intake, nutrient adsorption, mechanical stimulation of stomach wall and feeling of satiety. A novel approach to bariatric surgery is required, integrating competences in the areas of biomechanics, physiology and surgery, based on a strong interaction between engineers and clinicians. Preliminary results from coupled experimental and computational investigations are here reported. The analyses aim to develop computational tools for the investigation of stomach mechanical functionality in pre- and post-surgical conformations.


Impact of antimicrobial silver nanoparticles on anode respiring bacteria in a microbial electrolysis cell.
This study assessed the impact of antimicrobial AgNPs (50 mg L-1, 30-50 nm) on the electrocatalytic activity of a mixed-culture anode biofilm enriched with Geobacter species. The current densities and electrochemical kinetics were maintained after exposure to AgNPs in consecutive fed-batch cycles, despite significant changes in morphological structures and bacterial communities. Bacterial community analysis showed a substantial increase in the Geobacter population in response to AgNPs exposure, indicating their higher tolerance to AgNPs. In contrast, the population of other anode respiring bacteria (ARB) belongs to Acinetobacter, Dysgonomonas, and Cloacibacillus genera appeared to be very sensitive to AgNPs toxicity as their relative abundance significantly decreased. Microscopic imaging showed that AgNPs were accumulated within anode biofilm matrix without penetration inside the cells. Moreover, the anode biofilm became denser because of enhanced extracellular polymeric substances (EPSs) production by ARB after exposure of AgNPs, implying that EPS could protect ARB against AgNPs toxicity.


A menu-driven software package of Bayesian nonparametric (and parametric) mixed models for regression analysis and density estimation.
Most of applied statistics involves regression analysis of data. In practice, it is important to specify a regression model that has minimal assumptions which are not violated by data, to ensure that statistical inferences from the model are informative and not misleading. This paper presents a stand-alone and menu-driven software package, Bayesian Regression: Nonparametric and Parametric Models, constructed from MATLAB Compiler. Currently, this package gives the user a choice from 83 Bayesian models for data analysis. They include 47 Bayesian nonparametric (BNP) infinite-mixture regression models; 5 BNP infinite-mixture models for density estimation; and 31 normal random effects models (HLMs), including normal linear models. Each of the 78 regression models handles either a continuous, binary, or ordinal dependent variable, and can handle multi-level (grouped) data. All 83 Bayesian models can handle the analysis of weighted observations (e.g., for meta-analysis), and the analysis of left-censored, right-censored, and/or interval-censored data. Each BNP infinite-mixture model has a mixture distribution assigned one of various BNP prior distributions, including priors defined by either the Dirichlet process, Pitman-Yor process (including the normalized stable process), beta (two-parameter) process, normalized inverse-Gaussian process, geometric weights prior, dependent Dirichlet process, or the dependent infinite-probits prior. The software user can mouse-click to select a Bayesian model and perform data analysis via Markov chain Monte Carlo (MCMC) sampling. After the sampling completes, the software automatically opens text output that reports MCMC-based estimates of the model's posterior distribution and model predictive fit to the data. Additional text and/or graphical output can be generated by mouse-clicking other menu options. This includes output of MCMC convergence analyses, and estimates of the model's posterior predictive distribution, for selected functionals and values of covariates. The software is illustrated through the BNP regression analysis of real data.


Organic-Inorganic Nanocomposites via Self-Assembly of an Amphiphilic Triblock Copolymer Bearing a Poly(butadiene-g-POSS) Subchain in Epoxy Thermosets: Morphologies, Surface Hydrophobicity, and Dielectric Properties.
Organic-inorganic nanocomposites composed of polyhedral oligomeric silsesquioxane (POSS) and epoxy resin were prepared via self-assembly of an amphiphilic triblock copolymer bearing a poly(POSS) midblock in epoxy thermosets. First, this organic-inorganic amphiphilic triblock copolymer was synthesized via hydrosilylation of heptaphenylhydro POSS with an existing triblock copolymer containing a short polybutadiene midblock. It was found that this novel amphiphilic block copolymer can self-assemble into nanophases in epoxy thermosets. In the presence of preformed nanophases, the curing reaction was performed, and the organic-inorganic nanocomposites containing poly(POSS) microdomains were thus obtained. Compared with plain epoxy, the as-obtained thermosets exhibited enhanced surface hydrophobicity; the enhanced surface hydrophobicity is attributed to enrichment of the POSS component at the surface of the materials. Owing to the formation of poly(POSS) microdomains, the dielectric constants of the materials significantly reduced, whereas the dielectric loss remained almost unchanged.


Lipoid proteinosis of the oral mucosa: case report and review of the literature.
We describe a 37-year-old woman who presented with progressive mouth dryness. Physical examination revealed long-standing plaques on the face and upper limbs, papular lesions of the oral cavity and tongue firmness. A lower lip biopsy was performed. Light microscopy demonstrated accumulation of PAS-positive material around blood vessels, capillaries and salivary gland canaliculi as well as focally massive hyaline deposits in the submucosa. Immunohistochemistry revealed widespread presence of type IV collagen in the hyaline material and around thickened blood vessels. Laminin immunoreactivity was particularly strong at thickened basement membranes. The above findings were compatible with lipoid proteinosis, which is likely to involve primary perturbation of collagen metabolism and production of glycoproteins.


Quenched Hydrogen Exchange NMR of Amyloid Fibrils.
Amyloid fibrils are associated with a number of human diseases. These aggregatively misfolded intermolecular β-sheet assemblies constitute some of the most challenging targets in structural biology because to their complexity, size, and insolubility. Here, protocols and controls are described for experiments designed to study hydrogen-bonding in amyloid fibrils indirectly, by transferring information about amide proton occupancy in the fibrils to the dimethyl sulfoxide-denatured state. Since the denatured state is amenable to solution NMR spectroscopy, the method can provide residue-level-resolution data on hydrogen exchange for the monomers that make up the fibrils.


Ruminant field anesthesia.
Ruminants can be safely anesthetized in a field setting. The higher level of analgesia provided by anesthesia eliminates the need for local anesthetic blockade, which can prove useful when a procedure is expected to produce a substantial level of pain or local anesthetic blockade is not feasible. Certain aspects of anesthesia place the patient at greater risk than chemical restraint techniques. Knowledge and vigilance reduce the additional risks associated with anesthesia.


[Depression of penicillin-evoked seizure discharges of cerebrospinal neurons by magnesium ions].
In experiments on narcotized spinal cats perfusion of the lumbosacral segments of the spinal cord with penicillin (50 mmol/l) containing artificial cerebrospinal fluid led to the appearance of spontaneous negative small potentials in the dorsal roots and spontaneous repetitive bursts of impulses in the ventral roots of the perfused segments. The epileptogenic activity of penicillin was reduced or completely blocked if administration of the penicillin containing cerebrospinal fluid was preceded by 20-30 minute perfusion of the central canal of the lumbosacral segments with cerebrospinal fluid containing a high concentration of magnesium ions.


Custom-Made Quorum Sensing for a Eukaryote.
Quorum-sensing systems, common in prokaryotes, enable bacteria to coordinately regulate behavior with population density. Reporting recently in Cell Host & Microbe, Homer et al. (2016) characterize an elegant eukaryotic quorum-sensing pathway in the human pathogenic fungus Cryptococcus neoformans.


Nursing workload assessment in an intensive care unit: A retrospective observational study using the Nursing Activities Score.
Nursing Activities Score (NAS) is a promising tool for calculating the nursing workload in intensive care units (ICU). However, data on intensive care nursing activities in Portugal are practically non-existent. To assess the nursing workload in a Portuguese ICU using the NAS. Retrospective cohort study developed throughout the analysis of the electronic health record database from 56 adult patients admitted to a six-bed Portuguese ICU between 1 June-31 August 2020. The nursing workload was assessed by the Portuguese version of the NAS. The study was approved by the Hospital Council Board and Ethics Committee. The study report followed the STROBE guidelines. The average occupancy rate was 73.55% (±16.60%). The average nursing workload per participant was 67.52 (±10.91) points. There was a correlation between the occupancy rate and the nursing workload. In 35.78% of the days, the nursing workload was higher than the available human resources, overloading nurse staffing/team. The nursing workload reported follows the trend of the international studies and the results reinforce the importance of adjusting the nursing staffing to the complexity of nursing care in this ICU. This study highlighted periods of nursing workload that could compromise patient safety. This was one of the first studies carried out with the NAS after its cross-cultural adaptation and validation for the Portuguese population. The nursing workload at the patient level was higher in the first 24 h of ICU stays. Because of the 'administrative and management activities' related to the 'patient discharge procedures', the last 24 h of ICU stays also presented high levels of nursing workload. The implementation of a nurse-to-patient ratio of 1:1 may contribute to safer nurse staffing and to improve patient safety in this Tertiary (level 3) ICU.


Development of a Three-Dimensional Hand Model Using Three-Dimensional Stereophotogrammetry: Assessment of Image Reproducibility.
Using three-dimensional (3D) stereophotogrammetry precise images and reconstructions of the human body can be produced. Over the last few years, this technique is mainly being developed in the field of maxillofacial reconstructive surgery, creating fusion images with computed tomography (CT) data for precise planning and prediction of treatment outcome. Though, in hand surgery 3D stereophotogrammetry is not yet being used in clinical settings. A total of 34 three-dimensional hand photographs were analyzed to investigate the reproducibility. For every individual, 3D photographs were captured at two different time points (baseline T0 and one week later T1). Using two different registration methods, the reproducibility of the methods was analyzed. Furthermore, the differences between 3D photos of men and women were compared in a distance map as a first clinical pilot testing our registration method. The absolute mean registration error for the complete hand was 1.46 mm. This reduced to an error of 0.56 mm isolating the region to the palm of the hand. When comparing hands of both sexes, it was seen that the male hand was larger (broader base and longer fingers) than the female hand. This study shows that 3D stereophotogrammetry can produce reproducible images of the hand without harmful side effects for the patient, so proving to be a reliable method for soft tissue analysis. Its potential use in everyday practice of hand surgery needs to be further explored.


A prostaglandin D2 metabolite is elevated in the urine of Duchenne muscular dystrophy patients and increases further from 8 years old.
Duchenne muscular dystrophy (DMD) is a progressive muscle wasting disease caused by muscle dystrophin deficiency. Downstream of the primary dystrophin deficiency is not well elucidated. Here, the hypothesis that prostaglandin D2 (PGD2)-mediated inflammation is involved in the pathology of DMD was examined by measuring tetranor PGDM, a major PGD2 metabolite, in urine of DMD patients. We measured tetranor PGDM in urine using LC-MS/MS. First morning urine samples were collected from genetically confirmed DMD patients and age-matched healthy controls aged 4 to 15y. The urinary tetranor PGDM concentration was 3.08±0.15 and 6.90±0.35ng/mg creatinine (mean±SE) in 79 control and 191 DMD samples, respectively. The mean concentration was approximately 2.2-times higher in DMD patients than in controls (p<0.05). Remarkably, urinary tetranor PGDM concentrations in DMD patients showed chronological changes: it stayed nearly 1.5 times higher than in controls until 7y but surged at the age of 8y to a significantly higher concentration. Urinary tetranor PGDM concentrations were shown to be increased in DMD patients and became higher with advancing age. It was indicated that PGD2-mediated inflammation plays a role in the pathology of DMD.


Citizen science in monitoring food environments: a qualitative collective case study of stakeholders' experiences during the Local Environment Action on Food project in Alberta, Canada.
Citizen science bears potential to build a comprehensive view of global food environments and create a broader discussion about how to improve them. Despite its potential, citizen science has not been fully utilised in food environment research. Thus, we sought to explore stakeholders' experiences of the Local Environment Action on Food (LEAF) project, a community-based intervention that employs a citizen science approach to monitoring food environments. We used a qualitative collective case study design to explore citizen science through the LEAF process in seven communities in Alberta, Canada. Data generating strategies included semi-structured interviews with citizen scientists (n = 26), document review of communities' Mini Nutrition Report Cards (n = 7), and researcher observation. Data were analyzed in a multi-phase process, using Charmaz's constant comparison analysis strategy. Analysis revealed two main themes: relationship building and process factors. Communities used three interconnected strategies, engaging the right people, treading lightly, and reaching a consensus, to navigate the vital but challenging relationship building process. Process factors, which were influences on the LEAF process and relationship building, included the local context, flexibility in the LEAF process, and turnover among LEAF community groups. Citizen science through the LEAF project supported the creation and application of food environment evidence: it enabled residents to collect and interpret local food environment data, develop realistic recommendations for change, and provided them with an evidence-based advocacy tool to support the implementation of these recommendations. We recommend a web application that enables independent community food environment assessments. Such a tool could stimulate and sustain citizen involvement in food environment efforts, helping to build the necessary evidence base and promote the creation of healthy food environments.


Effects of coagulation conditions on structure and properties of cellulose-based fibers from aqueous NaOH solvent.
Hydroxyethyl cellulose (HEC) fibers with low degree of substitution were prepared by wet spinning of solutions in 8wt% NaOH solvent, and H2SO4/Na2SO4/ZnSO4 solution was chosen as coagulants. The influence of coagulation temperature and H2SO4 concentration on structure and mechanical properties of resultant HEC fibers was examined by synchrotron X-ray measurements, scanning electron microscope and tensile tests. The results were analyzed from the perspective of coagulation kinetics by determining coagulation rate and mass transfer rate difference. It is shown that the increase of either coagulation temperature or H2SO4 concentration would accelerate the diffusion and coagulation process. The lowest coagulation temperature of 20°C or moderate H2SO4 concentration around 10-15wt%, which provided a gentle solidification process, favored the development of regular crystal and oriented structure, uniform and dense inner structure as well as cross sections closer to circular, resulting in HEC fibers with better tensile properties.


Nutritional strategies for correcting low glucose values in patients with postbariatric hypoglycaemia: A randomized controlled three-arm crossover trial.
To evaluate the efficacy of nutritional hypoglycaemia correction strategies in postbariatric hypoglycaemia (PBH) after Roux-en-Y gastric bypass (RYGB). In a randomized, controlled, three-arm crossover trial, eight post-RYGB adults (mean [SD] 7.0 [1.4] years since surgery) with PBH ingested a solid mixed meal (584 kcal, 85 g carbohydrates, 21 g fat, 12 g protein) to induce hypoglycaemia on three separate days. Upon reaching plasma glucose of less than 3.0 mmol/L, hypoglycaemia was corrected with 15 g of glucose (G15), 5 g of glucose (G5) or a protein bar (P10, 10 g of protein) in random order. The primary outcome was percentage of time spent in the target plasma glucose range (3.9-5.5 mmol/L) during 40 minutes after correction. Postcorrection time spent in the target glucose range did not differ significantly between the interventions (P = .161). However, postcorrection time with glucose less than 3.9 mmol/L was lower after G15 than P10 (P = .007), whereas time spent with glucose more than 5.5 mmol/L, peak glucose and insulin 15 minutes postcorrection were higher after G15 than G5 and P10 (P < .001). Glucagon 15 minutes postcorrection was higher after P10 than after G15 and G5 (P = .002 and P = .003, respectively). G15 resulted in rebound hypoglycaemia (< 3.0 mmol/L) in three of eight cases (38%), while no rebound hypoglycaemia occurred with G5 and P10. Correcting hypoglycaemia with 15 g of glucose should be reconsidered in post-RYGB PBH. A lower dose appears to sufficiently increase glucose levels outside the critical range in most cases, and complementary nutrients (e.g. proteins) may provide glycaemia-stabilizing benefits. NTC05250271 (ClinicalTrials.gov).


Glucometrics in the first week of critical illness and its association with mortality.
Evaluation of glucometrics in the first week of ICU stay and its association with outcomes. Prospective observational study. Mixed ICU of teaching hospital. Adults initiated on insulin infusion for 2 consecutive blood glucose (BG) readings ≥180mg/dL. Glucometrics calculated from the BG of first week of admission: hyperglycemia (BG>180mg/dL) and hypoglycemia (BG<70mg/dL) episodes; median, standard deviation (SD) and coefficient of variation (CV) of BG, glycemic lability index (GLI), time in target BG range (TIR). Factors influencing glucometrics and the association of glucometrics to patient outcomes analyzed. A total of 5762 BG measurements in 100 patients of median age 55 years included. Glucometrics: hyperglycemia: 2253 (39%), hypoglycemia: 28 (0.48%), median BG: 169mg/dL (162-178.75), SD 31mg/dL (26-38.75), CV 18.6% (17.1-22.5), GLI: 718.5 [(mg/dL)2/h]/week (540.5-1131.5) and TIR 57% (50-67). Diabetes and higher APACHE II score were associated with higher SD and CV, and lower TIR. On multivariate regression, diabetes (p=0.009) and APACHE II score (p=0.016) were independently associated with higher SD. Higher SD and CV were associated with less vasopressor-free days; lower TIR with more blood-stream infections (BSI). Patients with higher SD, CV and GLI had a higher 28-day mortality. On multivariate analysis, GLI alone was associated with a higher mortality (OR 2.99, p=0.04). Glycemic lability in the first week in ICU patients receiving insulin infusion is associated with higher mortality. Lower TIR is associated with more blood stream infections.


DDC-induced amnesia and norepinephrine: a correlated behavioral-biochemical analysis.
Diethyldithiocarbamic acid (DDC), a dopamine-B-hydroxylase inhibitor, when injected into rats 30 min to 6 h before training of a passive avoidance task, impaired formation of long-term memory as indicated by performance on a retention test 24 h later. Performance of the task was at its minimum when injection occurred 2 to 4 h prior to training; recovery was evident in animals trained 5 or 6 h after drug treatment. Catecholamine assay of brains of temporally yoked animals showed that norepinephrine depletion followed a time course paralleling that of the amnesia. These findings support the hypothesis that the degree of memory storage, as reflected in performance following training in a passive avoidance task, can be directly correlated with the level of norepinephrine existing at the time of training.


Meeting report: Signal transduction meets systems biology.
In the 21st century, systems-wide analyses of biological processes are getting more and more realistic. Especially for the in depth analysis of signal transduction pathways and networks, various approaches of systems biology are now successfully used. The EU FP7 large integrated project SYBILLA (Systems Biology of T-cell Activation in Health and Disease) coordinates such an endeavor. By using a combination of experimental data sets and computational modelling, the consortium strives for gaining a detailed and mechanistic understanding of signal transduction processes that govern T-cell activation. In order to foster the interaction between systems biologists and experimentally working groups, SYBILLA co-organized the 15th meeting "Signal Transduction: Receptors, Mediators and Genes" together with the Signal Transduction Society (STS). Thus, the annual STS conference, held from November 7 to 9, 2011 in Weimar, Germany, provided an interdisciplinary forum for research on signal transduction with a major focus on systems biology addressing signalling events in T-cells. Here we report on a selection of ongoing projects of SYBILLA and how they were discussed at this interdisciplinary conference.


Optical investigation of the electron transfer protein azurin-gold nanoparticle system.
The hybrid system obtained by conjugating the protein azurin, which is a very stable and well-described protein showing a unique interplay among its electron transfer and optical properties, with 20-nm sized gold nanoparticles has been investigated. Binding of azurin molecules to gold nanoparticle surface results in the red shift of the nanoparticle resonance plasmon band and in the quenching of the azurin single tryptophan fluorescence signal. These findings together with the estimate of the hydrodynamic radius of the composite, obtained by means of Dynamic Light Scattering, are consistent with the formation of a monolayer of protein molecules, with preserved natural folding, on nanoparticle surface. The fluorescence quenching of azurin bound molecules is explained by an energy transfer from protein to metal surface and it is discussed in terms of the involvement of the Az electron transfer route in the interaction of the protein with the nanoparticle.


Health threat of PM2.5-bound trace elements exposure on asthma hospital admission: A time-stratified case-crossover study.
Environmental pollution of trace elements has become of main concern due to the adverse effects. To estimate the impact of PM2.5-bound trace elements on human health, a time-stratified case-crossover study has been designed to examine the short-term associations between 28 elements and asthma hospitalizations from January 2019 to November 2021 in Xiamen, China. This research summarized the major components that pose health risks in different seasons and took risk assessment for different groups. We found that an inter-quartile range (IQR) increase of Ca, Fe, Mn, Pd, Si, and Ti was positively associated with the incidence of asthma in the lag of 0-4 days. In winter, the elements that predisposed the population to asthma attack were the most, such as Al, Ba, Ca, Cr, Cu, Fe, K, Mn, Pd, Si, and Ti, and these elements are at higher risk for longer periods of time. The discrepancy in risk levels and major elements of asthma diagnosis among various age groups were also found in this work. Our results provided insights into the development of specific policies to reduce the risk of asthma attacks due to exposure to PM2.5-bound trace elements.


Molecular Design: Network Architecture and Its Impact on the Organization and Mechanics of Peptide-Polyurea Hybrids.
Nature has achieved controlled and tunable mechanics via hierarchical organization driven by physical and covalent interactions. Polymer-peptide hybrids have been designed to mimic natural materials utilizing these architectural strategies, obtaining diverse mechanical properties, stimuli responsiveness, and bioactivity. Here, utilizing a molecular design pathway, peptide-polyurea hybrid networks were synthesized to investigate the role of architecture and structural interplay on peptide hydrogen bonding, assembly, and mechanics. Networks formed from poly(β-benzyl-l-aspartate)-poly(dimethylsiloxane) copolymers covalently cross-linked with a triisocyanate yielded polyurea films with a globular-like morphology and parallel β-sheet secondary structures. The geometrical constraints imposed by the network led to an increase in peptide loading and ∼7x increase in Young's modulus while maintaining extensibility (∼160%). Thus, the interplay of physical and chemical bonds allowed for the modulation of resulting mechanical properties. This investigation provides a framework for the utilization of structural interplay and mechanical tuning in polymer-peptide hybrids, which offers a pathway for the design of future hybrid biomaterial systems.


The feeding biology of a species-rich genus of rainforest grasshoppers (Rhachicreagra: Orthoptera, Acrididae) : I. Foodplant use and foodplant acceptance.
1. 11 Costa Rican species of the forest light-gap grasshopper genus Rhachicreagra were shown by direct observation and by faecal analysis to be each narrow-range disjunct oligophages, eating typically only 3-6 species from the hundreds present in their habitat. The diet of the different species varies considerably, some pairs showing no overlap, others having several species in common. 2. All Rhachicreagra spp. accepted in captivity any plant found in the diet of any other congeneric species, but refused almost all other plants. There is thus a generic spectrum of acceptable plants. It includes one to several members of each of 7 families: Compositae, Urticaceae, Umbellifereae, Amaranthaceae, Phytolaccaceae, Gramineae and an unidentified monocotyledenous family. Within this range, the diet of any given species or population appears to be determined primarily by availability within the habitat. There is no evidence for the hypothesis that food-plant shifts are directly associated with speciation within the genus. 3. Compositae are a component of the diet of most species, and include the principal (=most used) foodplant of many, including the morphologically most primitive species. In lowland habitats, Urticaceae supplement or replace the Compositae in the diet, in montane habitats Hydrocotyle (Umbelliferae) is important. Phytolacca and Iresine (Amaranth.) are usually minor constituents of the diet of lowland species, but Iresine is the principal foodplant of Rh. obsidian. The monocotyledenous families appear to be eaten only when other footplants are in short supply. The literature suggests that the dicotyledenous families favoured are listinguished by their relatively high content of nitrogen and cations. They are also variously rich in saponins, flavonoids, alkaloids, polyacetylenes and sesquiterpene lactones. 4. The present ecological distribution of the genus is wider han that of any single known foodplant, and appears to be made possible by the diversity of the plants accepted; these include species typical of lowland and montane habitats and of drier and wetter climates.


Osteoporosis in primary biliary cirrhosis: pathogenesis and treatment.
Osteoporosis, characterized by loss of bone strength leading to fragility fractures, is a common event in patients who have primary biliary cirrhosis. Although its pathogenesis is not well known, it results mainly from low bone formation. There is no specific treatment, but bisphosphonates, especially alendronate, effectively increases bone mass and prevents bone loss. Despite these favorable effects on bone mass, no clear effects on decreasing the fracture rate are demonstrated, probably because of the low number of patients included in the trials. The potential usefulness of new agents requires further evaluation.


Prognosis in acute pancreatitis associated with HIV infection.
This prospective study compared scoring systems in predicting adverse outcomes in HIV associated acute pancreatitis (HIV+ve AP) METHODS: Systemic inflammatory response syndrome (SIRS), Glasgow criteria, C-reactive protein (CRP), bedside index of severity in acute pancreatitis (BISAP) and APACHE II scores using standard cut-off values were used to predict the endpoint of moderate and severe disease in HIV-ve and HIV+ve patients and in CD4 counts above and below 200 cells/mm3. Ninety (38%) of 238 patients with AP were HIV+ve. Fifteen had organ failure, 33 local complications and 12 patients died. Advanced age was not associated with severe disease. The APACHE II was the best predictor of severe disease in HIV+ve (AUC 0.88) and HIV-ve patients (AUC 0.81) and CRP was the poorest predictor (AUC 0.59) in HIV+ve patients. In HIV+ve patients with CD4 counts greater and less than 200 cells/mm3 the Glasgow and APACHE II scores were the best prognosticators (AUC > 0.8) and BISAP in patients with CD4 > 200 cells/mm3 (AUC 0.90). The APACHE II score was most effective irrespective of HIV status whereas the BISAP scores was better in CD4 > 200 cells/mm3.


A novel salting-out assisted extraction coupled with HPLC- fluorescence detection for trace determination of vitamin K homologues in human plasma.
Recently, new physiological roles of vitamin K homologues have been established in the treatment of rheumatoid arthritis, osteoporosis, hepatocellular carcinoma and leukemia. However, relatively high plasma protein binding, low plasma concentrations and occurrences of interfering lipids make accurate determination of vitamin K homologues a challenging task. Therefore, a sensitive and reliable salting-out assisted liquid/liquid extraction (SALLE) method coupled with HPLC-Fluorescence detection was designed for efficient extraction and quantification of trace levels of vitamin K homologues in human plasma. The investigated vitamin K homologues were phylloquinone (PK, vitamin K1), menaquinone-4 (MK-4) and menaquinone-7 (MK-7). The method employed a new efficient fluorescence derivatization reaction using ethanolic solution of stannous chloride in acidic solution to generate highly fluorescent naphthohydroquinone derivatives. Correlation coefficients were more than 0.998 in the concentration ranges of 0.3-100 ng mL(-1) with detection limits of 0.1-0.17 ng mL(-1) in human plasma. The developed HPLC-FL system was successfully applied for sensitive determination of vitamin K homologues in plasma of healthy volunteers. The developed method may provide a valuable tool in the pharmacoinformatic studies concerning the roles of vitamin K homologues.


A ventilation-filtration unit for respiratory isolation.
The development of a new method for achieving respiratory isolation in hospitals, clinics, and residential facilities, in response to the increasing risk of transmission of tuberculosis and the limitations of the currently available isolation systems. Ultraviolet (UV) light and ultra-low-penetration air filtration were combined with a ventilation unit and adapted for use in modular isolation rooms or for conversion of existing rooms. The ventilation-filtration unit efficiently cleared bacterial aerosols and particles > 0.2 microns from the air, maintained required negative pressures and airflows, and provided directional airflow within rooms.


Culture of human glomerular cells.
Human glomeruli were routinely cultured in Waymouth's medium supplemented with insulin and conditioned medium. Three cell types were seen in the culture of both adult and infant kidneys, but the morphology of the glomerular cellular outgrowths depended on the age of the patient from which the kidney was obtained. Cultures of glomeruli from older individuals resulted in more "differentiated" cells, but both adult and infant glomerular cells rapidly became "dedifferentiated" as the length of time in culture increased. Outgrowths of cultured glomeruli did not contain fibroblasts or tubular cells. Finally, synthesis of basement membrane material by these cultured glomerular cells was demonstrated.


High-sensitive troponin T and N-terminal-brain-natriuretic-peptide predict outcome in symptomatic aortic stenosis.
Aortic stenosis (AS) and atherosclerosis share similarities when it comes to risk factors and disease progression. Like in other heart diseases, we hypothesized that biomarkers like high-sensitive troponin T (hsTnT), N-terminal-pro-brain-natriuretic-peptide (NT-proBNP) and high-sensitive C-reactive protein (hsCRP) could be useful in risk stratification. A total of 136 patients (57% men, mean age 74 years), referred for evaluation of AS (valve area 0.62 cm(2), left ventricular ejection fraction 64%) were consecutively enrolled in the study. The relationship between hsTnT, hsCRP and NT-proBNP, different echocardiographic parameters of AS and cardiac function were investigated as well as their relation to all-cause mortality. In contrast to hsCRP, hsTnT and NT-proBNP were individually correlated with prognosis. Regression analysis identified diabetes and the combination of hsTnT and NT-proBNP as significant predictors of all-cause mortality. When analyzing patients without surgery separately, only the combination of hsTnT and NT-proBNP were identified as a significant predictor of all-cause mortality in multivariable analysis. The combination of NT-proBNP and hsTnT came out as the strongest predictor of outcome irrespective of surgical treatment or not and could be of particular interest in risk-stratification in AS-patients. The results should be confirmed in prospective studies both in symptomatic and asymptomatic patients.