Use-dependent activation of neuronal Kv1.2 channel complexes.
In excitable cells, ion channels are frequently challenged by repetitive stimuli, and their responses shape cellular behavior by regulating the duration and termination of bursts of action potentials. We have investigated the behavior of Shaker family voltage-gated potassium (Kv) channels subjected to repetitive stimuli, with a particular focus on Kv1.2. Genetic deletion of this subunit results in complete mortality within 2 weeks of birth in mice, highlighting a critical physiological role for Kv1.2. Kv1.2 channels exhibit a unique property described previously as "prepulse potentiation," in which activation by a depolarizing step facilitates activation in a subsequent pulse. In this study, we demonstrate that this property enables Kv1.2 channels to exhibit use-dependent activation during trains of very brief depolarizations. Also, Kv subunits usually assemble into heteromeric channels in the central nervous system, generating diversity of function and sensitivity to signaling mechanisms. We demonstrate that other Kv1 channel types do not exhibit use-dependent activation, but this property is conferred in heteromeric channel complexes containing even a single Kv1.2 subunit. This regulatory mechanism is observed in mammalian cell lines as well as primary cultures of hippocampal neurons. Our findings illustrate that use-dependent activation is a unique property of Kv1.2 that persists in heteromeric channel complexes and may influence function of hippocampal neurons.


Hepatitis C-associated hepatocellular carcinomas in non-cirrhotic livers.
Chronic hepatitis C viral infection can lead to cirrhosis and hepatocellular carcinoma. It is generally believed that hepatitis C infection is not oncogeneic per se, but that the presence of cirrhosis determines the increased risk for hepatocellular carcinoma. However, a search of surgical pathology files from two large tertiary care centers for the years 2001-2008 identified a total of 18 hepatocellular carcinomas in non-cirrhotic livers with chronic hepatitis C infection. In six cases the background livers showed bridging fibrosis, while the remainder showed lower stages of fibrosis. Cases were negative for clinical and serological evidence of hepatitis B co-infection, and occult hepatitis B test was negative by PCR of formalin-fixed, paraffin embedded tissues. The tumors were also negative for TP53, exon 7, codon 249 mutations, a characteristic mutation strongly linked to aflatoxin exposure. The hepatocellular carcinomas had typical histology with no enrichment for unusual growth patterns or histological features. Among all resected hepatocellular carcinomas in non-cirrhotic livers over this time period, the prevalence of 16% with HCV infection was significantly greater than that expected by chance. In conclusion, these results demonstrate that hepatocellular carcinomas can arise in livers chronically infected with hepatitis C but without cirrhosis. These findings raise the possibility that in some cases hepatitis C infection and inflammation can be directly oncogeneic. It is also possible that established cirrhosis may have regressed in some cases. Regardless of the mechanism, these findings highlight an important and previously under-recognized risk for hepatocellular carcinoma in HCV-infected individuals who do not have cirrhosis.


How to recognize and manage psychological distress in cancer patients.
Psychological distress is common in cancer patients, however, it is often unrecognized and untreated. We aimed to identify barriers to cancer patients expressing their psychological concerns, and to recommend strategies to assist oncologists to elicit, recognize, and manage psychological distress in their patients. Medline, Psychlit, and the Cochrane databases were searched for articles relating to the detection of emotional distress in patients. Patients can provide verbal and non-verbal information about their emotional state. However, many patients may not reveal emotional issues as they believe it is not a doctor's role to help with their emotional concerns. Moreover, patients may normalize or somatize their feelings. Anxiety and depression can mimic physical symptoms of cancer or treatments, and consequently emotional distress may not be detected. Techniques such as active listening, using open questions and emotional words, responding appropriately to patients' emotional cues, and a patient-centred consulting style can assist in detection. Screening tools for psychological distress and patient question prompt sheets administered prior to the consultation can also be useful. In conclusion, the application of basic communication techniques enhances detection of patients' emotional concerns. Training oncologists in these techniques should improve the psychosocial care of cancer patients.


Thalidomide for erythema nodosum leprosum and other applications.
Thalidomide, administered as a sedative and antiemetic decades ago, was considered responsible for numerous devastating cases of birth defects and consequently was banned from markets worldwide. However, the drug remarkably has resurfaced with promise of immunomodulatory benefit in a wide array of immunologic disorders for which available treatments were limited. It is approved by the Food and Drug Administration for erythema nodosum leprosum (ENL). Although the relative paucity of leprosy and ENL worldwide may perceivably limit interest in and knowledge about thalidomide, increasing numbers of new and potential uses expand its applicability widely beyond ENL. Thalidomide, an inhibitor of tumor necrosis factor a, is the best known agent for short-term treatment of ENL skin manifestations, as well as postremission maintenance therapy to prevent recurrence. For this indication, it is effective as monotherapy and as part of combination therapy with corticosteroids. Studies of thalidomide in chronic graft-versus-host disease showed benefit in children and adults as treatment, but not as prophylaxis. The agent has been administered successfully for treatment of cachexia related to cancer, tuberculosis, and human immunodeficiency virus infection, although evidence of efficacy is inconclusive. Thalidomide monotherapy effectively induced objective response in trials in patients with both newly diagnosed and advanced or refractory multiple myeloma. Combination therapy with thalidomide and corticosteroids was also effective in these patients, as well as in treatment of aphthous and genital ulcers. Limited evidence supports the drug's benefit in treatment of Kaposi's sarcoma. Other thalidomide applications include Crohn's disease, rheumatoid arthritis, and multiple sclerosis. Somnolence, constipation, and rash were the most frequently cited adverse effects in studies, but thalidomide-induced neuropathy and idiopathic thromboembolism were critical causes for drug discontinuation. Thalidomide is still contraindicated in pregnant women, women of childbearing age, and sexually active men not using contraception. Clinicians should be conversant with thalidomide in ENL (its primary application) in the natural course of leprosy, as well as in the agent's other applications.


Effects of natal habitat odour, reinforced by adult experience, on choice of oviposition site in the mosquito Aedes aegypti.
The effects of natal experience on the oviposition behaviour of adult female mosquitoes were investigated in the laboratory using Aedes aegypti (L.) (Diptera: Culicidae). 'Treatment' mosquitoes were exposed to a dilute repellent (inducing stimulus) in their breeding water (aquatic stages) and/or in the air (adults) during various combinations of life stages [larval only (L regime); larval and pupal (LP regime); larval, pupal and emergent adult (LPE regime); larval, pupal, emergent adult and adult (LPEA regime); pupal, emergent adult and adult (PEA regime); adult only (A regime)]. 'Control' mosquitoes were raised in an identical manner, but were not exposed to the inducing stimulus. The oviposition behaviour of treatment and control females was assessed in an oviposition assay that presented a choice of water with or without the inducing stimulus. Of the 435 mosquitoes tested in the experiment, 176 were non-distributors (i.e. laid all of their eggs in only one of the choices). Treatment females (distributors plus non-distributors) reared in the presence of the inducing stimulus throughout their lives (LPEA regime) showed a significant preference for the oviposition option containing the inducing stimulus (24/36 females) compared with corresponding controls (5/39 females). Distributors reared under the LPEA and PEA regimes also showed this preference (6/6 treatment vs. 2/29 control females, and 13/18 treatment vs. 7/23 control females, respectively). Females that had been exposed to the inducing stimulus as either immatures or adults only showed no preference for, and some showed an aversion to, the treatment oviposition option. This is interpreted as evidence for a natal habitat preference induction (NHPI) in this species, albeit one that requires extensive reinforcement in the adult stage. This adult experience-reinforced NHPI (AER-NHPI) is discussed in terms of its adaptive significance for container breeders, the possible timing mechanism and sensory basis of induction and potential practical consequences.


Modelling Subarctic watershed dissolved organic carbon response to hydroclimatic regime.
Shifts in hydroclimatic regimes associated with global climate change may impact freshwater availability and quality. In high latitudes of the northern hemisphere, where vast quantities of carbon are stored terrestrially, explaining landscape-scale carbon (C) budgets and associated pollutant transfer is necessary for understanding the impact of changing hydroclimatic regimes. We used a dynamic modelling approach to simulate streamflow, DOC concentration, and DOC export in a northern Canadian catchment that has undergone notable climate warming, and will continue to for the remainder of this century. The Integrated Catchment model for Carbon (INCA-C) was successfully calibrated to a multi-year period (2012-2016) that represents a range in hydrologic conditions. The model was subsequently run over 30-year periods representing baseline and two future climate scenarios. Average discharge is predicted to decrease under an elevated temperature scenario (22-27 % of baseline) but increase (116-175 % of baseline) under an elevated temperature and precipitation scenario. In the latter scenario the nival hydroclimatic regime is expected to shift to a combined nival and pluvial regime. Average DOC flux over 30 years is predicted to decrease (24-27 % of baseline) under the elevated temperature scenario, as higher DOC concentrations are offset by lower runoff. Under the elevated temperature and precipitation scenario, results suggest an increase in carbon export of 64-81 % above baseline. These increases are attributed to greater connectivity of the catchment. The largest increase in DOC export is expected to occur in early winter. These predicted changes in DOC export, particularly under a climate that is warmer and wetter could be part of larger ecosystem change and warrant additional monitoring efforts in the region.


A systematic review of the nutritional implications of military rations.
The objective of this study was to assess military rations. Military rations must provide military personnel with suitable nutrition, as high-quality nutrition is crucial for promoting health. This systematic review provides information regarding the nutritional design of military rations according to energy and protein intake and supplements. A systematic search was conducted for articles published to date regarding the nutritional needs involved in a military diet for an active population of either gender, ranging from 18 to 60 years old, without excluding material by year, type of document, location, or author. The pertinent articles found were published from 1994 to 2017. Physical activity and environmental factors directly affect calculating energy needs and carbohydrate intake when designing military rations. However, the results showed no influence on protein, fat and/or sodium intake. Furthermore, the search revealed that military populations have a high intake of dietary supplements; it is thus worth considering their incorporation into campaign rations. Military personnel macro- and micro-nutrient needs depend on physical activity and nutrient intake measurement methodologies. The data regarding military personnel dietary supplement consumption indicated that this is a niche worth considering when designing military rations. Research on nutritional needs for military rations could concentrate on optimizing the amount of macro- and micro-nutrients and their biological value, according to physical activity, as well as maintaining their sensory quality, safety and shelf life using different processing technologies.


Aberrant cytokinesis and cell fusion result in multinucleation in HepG2 cells exposed to silica nanoparticles.
The multinucleation effect of silica nanoparticles (SiNPs) had been determined in our previous studies, but the relative mechanisms of multinucleation and how the multinucleated cells are generated were still not clear. This extensional study was conducted to investigate the mechanisms underlying the formation of multinucleated cells after SiNPs exposure. We first investigated cellular multinucleation, then performed time-lapse confocal imaging to certify whether the multinucleated cells resulted from cell fusion or abnormal cell division. Our results confirmed for the first time that there are three patterns contributing to the SiNPs-induced multinucleation in HepG2 cells: cell fusion, karyokinesis without cytokinesis, and cytokinesis followed by fusion. The chromosomal passenger complex (CPC) deficiency and cell cycle arrest in G1/S and G2/M checkpoints may be responsible for the cell aberrant cytokinesis. The activated MAPK/ERK1/2 signaling and decreased mitosis related proteins might be the underlying mechanism of cell cycle arrest and thus multinucleation. In summary, we confirmed the hypothesis that aberrant cytokinesis and cell fusion resulted in multinucleation in HepG2 cells after SiNPs exposure. Since cell fusion and multinucleation were involved in genetic instability and tumor development, this study suggests the potential ability of SiNPs to induce cellular genetic instability. These findings raise concerns with regard to human health hazards and environmental risks with SiNPs exposure.


Excited-State Proton Transfer to H2O in Mixtures of CH3CN-H2O of a Superphotoacid, Chlorobenzoate Phenol Cyanine Picolinium (CBCyP).
Steady-state and time-resolved fluorescence techniques were employed to study a superphotoacid with a p Ka* of ∼-7, the chlorobenzoate phenol cyanine picolinium salt (CBCyP) in acetonitrile-water mixtures. We found that the time-resolved fluorescence is bimodal. The amplitude of the short-time component depends on χwater; the larger χwater, the greater the amplitude. We found that the excited-state proton-transfer (ESPT) rate constant, kPT, is ≥5 × 1012 s-1 in mixtures of χwater ≥ 0.08, whereas in neat water, kPT = 6 × 1012 s-1. The long-time component has a lifetime of 50 ps at χwater = 0.75. We attribute this time component to the CBCyP molecules that are not hydrogen-bonded to H2O clusters. The results suggest that the ESPT rate constant to water in acetonitrile-water mixtures depends only slightly on the water cluster size and structure surrounding the CBCyP molecule. We attribute the independence of the ESPT rate on the average water-cluster size to the large photoacidity of CBCyP. QM TD-DFT calculations found that in the excited-state the RO-(S1) species that is formed by the ESPT process is more stable than the ROH(S1) species by -5 kcal/mol when four water molecules accept the proton, and when six water molecules accept the proton, the RO-(S1) drops to -10 kcal/mol. The calculations show that energy stabilities are kept constant in implicit CH3CN-H2O solvent mixtures of dielectric constant of ε ≥ 45.


Estimated glomerular filtration rate by serum cystatin C correlates with cardiometabolic parameters in patients with primary hyperparathyroidism.
Patients with primary hyperparathyroidism (PHPT) are at risk of chronic kidney disease (CKD). Cystatin C (Cys-C) is considered a more reliable tool to assess glomerular filtration rate (GFR) than creatinine. The study aimed to assess circulating Cys-C and its relationships with biochemical PHPT and cardiometabolic parameters. The present cross-sectional study was performed in academic endocrine units on PHPT patients (n=190) and non-hypertensive, non-diabetic, age- and sex-matched healthy controls (n=135) with no established CKD. The main outcomes were creatinine by alkaline picrate method, Cys-C by immunonephelometry and calculation of estimated GFR based on creatinine and Cys-C (eGFRcr-cys) using the CKD-EPI equation. In PHPT patients, circulating Cys-C ranged 0.45-3.13 mg/l and correlated with creatinine, age and BMI. Mean Cys-C level was higher in PHPT patients than in controls (0.93±0.02 vs 0.78±0.14 mg/l; P=0.03). Cys-C levels in PHPT patients were predicted by age, BMI, ionized calcium, hypertension and HDL-cholesterol, the most significant determinant being ionized calcium. Cys-C positively correlated with cardiovascular disease (CVD) occurrence. Overall, 18.4% of PHPT patients with eGFRcr >60 ml/min per 1.73 m(2) (n=169) had Cys-C levels higher than the 95th percentile in controls (1.03 mg/l), consistent with a preclinical CKD, which was associated with hypertension and insulin resistance. Considering eGFRcr-cys, CKD (stages G3a, G3b, 4) was diagnosed in 13.7% of PHPT patients. Estimated GFRcr-cys, but not eGFR based on creatinine, was predicted by insulin resistance and hypertension and positively correlated with CVD. Elevated Cys-C levels were associated with ionized calcium, cardiometabolic risk factors and CVD, and identified preclinical CKD in PHPT patients.


Remission and Low Disease Activity Status (LDAS) protect lupus patients from damage occurrence: data from a multiethnic, multinational Latin American Lupus Cohort (GLADEL).
To evaluate disease activity statuses' (DAS') impact on systemic lupus erythematosus (SLE) outcomes. Four DAS were defined: remission off-therapy: SLE Disease Activity Index (SLEDAI)=0, no prednisone or immunosuppressive drugs (IS); remission on-therapy: SLEDAI=0, prednisone ≤5 mg/day and/or IS (maintenance); low (L) DAS: SLEDAI ≤4, prednisone ≤7.5 mg/day and/or IS (maintenance); non-optimally controlled: SLEDAI >4 and/or prednisone >7.5 mg/day and/or IS (induction). Antimalarials were allowed in all. Predefined outcomes were mortality, new damage (increase of at least one Systemic Lupus International Collaborating Clinics/American College of Rheumatology (SLICC/ACR) damage index (SDI) point) and severe new damage (increase of at least 3 SDI points). Univariable and multivariable Cox regression models were performed to define the impact of DAS, as time-dependent variable, on these outcomes. 1350 patients were included, 79 died during follow-up, 606 presented new and 177 severe new damage. In multivariable analyses, remission (on/off-therapy) was associated with a lower risk of new (HR 0.60; 95% CI 0.43 to 0.85), and of severe new damage (HR 0.32; 95% CI 0.15 to 0.68); low disease activity status (LDAS) was associated with a lower risk of new damage (HR 0.66; 95% CI 0.48 to 0.93) compared with non-optimally controlled. No significant effect on mortality was observed. Remission was associated with a lower risk of new and severe new damage; LDAS with a lower risk of new damage after adjusting for other damage confounders.


Protein polymorphisms, segregation in genetic crosses and genetic distances among fishes of the genus Xiphophorus (Poeciliidae).
The products of 49 protein-coding loci were examined by starch gel electrophoresis for populational variation in six species of Xiphophorus fishes and/or segregation in intra-and interspecific backcross and intercross hybrids. Electrophoretic variation was observed for 29 of the 35 locus products in a survey of 42 population samples. The highest frequency of polymorphic loci observed in noninbred populations was 0.143. After ten or more generations of inbreeding, all loci studied were monomorphic. Inbred strains generally exhibited the commonest electrophoretic alleles of the population from which they were derived. An assessment of genetic distances among Xiphophorus populations reflected classical systematic relationships and suggested incipient subspeciation between X. maculatus from different drainages as well as several species groups. Thirty-three loci were analyzed with respect to segregation in hybrids. The goodness of fit of segregations to Mendelian expectations at all loci analyzed (except loci in linkage group I) is interpreted as evidence for high genetic compatibility of the genomes of Xiphophorus species. It is anticipated that these data will result in a rapid expansion of the assignment of protein-coding loci to linkage groups in these lower vertebrate species.


Platelet factor 4 and the platelet secreted proteoglycan: immunologic characterization by crossed immunoelectrophoresis.
Platelet factor 4 (PF4) is a hydrophobic, alpha-granule protein with potent antiheparin activity. It also binds to a chondroitin sulfate-containing proteoglycan (PG) isolated from platelets. In order to evaluate further the relationship between PF4 and the chondroitin sulfate-containing proteoglycan in resting platelets, the PF4-binding proteoglycan from human platelets has been purified using purified PF4 as an affinity ligand and used to prepare polyclonal antiserum. Two antisera have been characterized: one reacts primarily with chondroitin sulfate (CS), the other reacts with the protein core of the platelet proteoglycan after chondroitinase AC digestion. PF4 and PG core protein antigen are present in separate, dissimilar precipitin arcs when triton-solubilized platelets are analyzed by crossed immunoelectrophoresis using polyclonal antisera to purified PF4 and PG. PF4 was demonstrated in a complex with a separate chondroitin sulfate antigen by crossed immunoelectrophoresis (CIE) experiments in which either anti-PF4 or anti-CS antisera was incorporated in the intermediate gel. Both the PF4-chondroitin sulfate complex and the proteoglycan are secreted from platelets when fresh, washed human platelets are stimulated by human alpha-thrombin. This second antigen may represent the PG after posttranslational modification of a precursor form of the proteoglycan.


Understanding the pathways between the use of urban green spaces and self-rated health: A case study in Mexico City.
In recent decades, extensive research has demonstrated the positive impact of urban green spaces (UGS) on public health through several pathways. However, in the context of Latin America, particularly Mexico City, there remains a notable scarcity of evidence linking UGS use to health outcomes and an insufficient understanding of the pathways or factors underlying these associations. Therefore, this study employs Structural Equation Modeling (SEM) to investigate the intricate pathways between UGS use and residents' perceived health in Mexico City, a densely populated urban center. The SEM integrates three key mediators: sentiments towards UGS, UGS quality, and time spent within these spaces. Survey data was collected through an online survey distributed via social media in May 2020 (n = 1,707). The findings indicate a minor yet significant direct link between UGS use and self-reported health (0.0427, p < 0.1). Conversely, the indirect pathways through sentiments towards UGS, UGS quality, and time spent in UGS were highly significant (0.1950, p < 0.01), underscoring their substantial role as mediators in the UGS use-health association. While a comprehensive understanding of the mechanisms linking perceived health to UGS use in Mexico City requires further research, this study proposes that fostering positive sentiments towards UGS, enhancing UGS quality, and encouraging extended visits to green areas could potentially amplify the perceived health benefits associated with UGS use among residents. These insights offer valuable inputs for policymaking, emphasizing the importance of integrating public perspectives to optimize nature-based solutions and broaden their positive impact within Mexico City.


Flash survey on severe acute respiratory syndrome coronavirus-2 infections in paediatric patients on anticancer treatment.
Since the beginning of COVID-19 pandemic, it is known that the severe course of the disease occurs mostly among the elderly, whereas it is rare among children and young adults. Comorbidities, in particular, diabetes and hypertension, clearly associated with age, besides obesity and smoke, are strongly associated with the need for intensive treatment and a dismal outcome. A weaker immunity of the elderly has been proposed as a possible explanation of this uneven age distribution. Thus, there is concern that children treated for cancer may allso be at risk for an unfavourable course of infection. Along the same line, anecdotal information from Wuhan, China, mentioned a severe course of COVID-19 in a child treated for leukaemia. We made a flash survey on COVID-19 incidence and severity among children on anticancer treatment. Respondents were asked by email to fill in a short Web-based survey. We received reports from 25 countries, where approximately 10,000 patients at risk are followed up. At the time of the survey, more than 200 of these children were tested, nine of whom were positive for COVID-19. Eight of the nine cases had asymptomatic to mild disease, and one was just diagnosed with COVID-19. We also discuss preventive measures that are in place or should be taken and treatment options in immunocompromised children with COVID-19. Thus, even children receiving anticancer chemotherapy may have a mild or asymptomatic course of COVID-19. While we should not underestimate the risk of developing a more severe course of COVID-19 than that observed here, the intensity of preventive measures should not cause delays or obstructions in oncological treatment.


Validation data-based adjustments for outcome misclassification in logistic regression: an illustration.
Misclassification of binary outcome variables is a known source of potentially serious bias when estimating adjusted odds ratios. Although researchers have described frequentist and Bayesian methods for dealing with the problem, these methods have seldom fully bridged the gap between statistical research and epidemiologic practice. In particular, there have been few real-world applications of readily grasped and computationally accessible methods that make direct use of internal validation data to adjust for differential outcome misclassification in logistic regression. In this paper, we illustrate likelihood-based methods for this purpose that can be implemented using standard statistical software. Using main study and internal validation data from the HIV Epidemiology Research Study, we demonstrate how misclassification rates can depend on the values of subject-specific covariates, and we illustrate the importance of accounting for this dependence. Simulation studies confirm the effectiveness of the maximum likelihood approach. We emphasize clear exposition of the likelihood function itself, to permit the reader to easily assimilate appended computer code that facilitates sensitivity analyses as well as the efficient handling of main/external and main/internal validation-study data. These methods are readily applicable under random cross-sectional sampling, and we discuss the extent to which the main/internal analysis remains appropriate under outcome-dependent (case-control) sampling.


[The effect of intravascular hemolysis on the body of rheumatics with heart valve prostheses].
The rate of intravascular hemolysis and associated complications was studied in 100 patients with rheumatic heart disease within 2 to 20 years after replacement of the heart valves. Hemolysis of varying intensity was recorded in 72% of patients, anemia in 5%, hemosiderinuria in 27%, cholelithiasis in 4%, and sideropenia in 36.8% of the examined. Renal failure was diagnosed in none of the patients. Thromboembolic complications were more frequently encountered in patients with hemolysis. In order to prevent anemia and sideropenia, it is recommended that control may be exercised over hemolysis and iron content in the blood of patients with prosthetic heart valves.


Two Methods of Monitoring Cats at a Landscape-Scale.
Feral cats are difficult to manage and harder to monitor. We analysed the cost and the efficacy of monitoring the pre- and post-bait abundance of feral cats via camera-traps or track counts using four years of data from the Matuwa Indigenous Protected Area. Additionally, we report on the recovery of the feral cat population and the efficacy of subsequent Eradicat® aerial baiting programs following 12 months of intensive feral cat control in 2019. Significantly fewer cats were captured in 2020 (n = 8) compared to 2019 (n = 126). Pre-baiting surveys for 2020 and 2021 suggested that the population of feral cats on Matuwa was very low, at 5.5 and 4.4 cats/100 km, respectively, which is well below our target threshold of 10 cats/100 km. Post-baiting surveys then recorded 3.6 and 3.0 cats/100 km, respectively, which still equates to a 35% and 32% reduction in cat activity. Track counts recorded significantly more feral cats than camera traps and were cheaper to implement. We recommend that at least two methods of monitoring cats be implemented to prevent erroneous conclusions.


Microbial fermentation of rice straw: nutritive composition and in vitro digestibility of the fermentation products.
Rice straw was fermented with Cellulomonas sp. and Alcaligenes faecalis. Microbial cells and undigested residue, as well as chemically treated (NaOH or NH4OH) and untreated straws, were analyzed for nutrient composition and in vitro digestibility. In a typical fermentation, 75% of the rice straw substrate was digested, and 18.6% of the total substrate weight that disappeared was recovered as microbial protein. The microbial cell fraction was 37% protein and 5% crude fiber; the residue was 12% protein and 45% crude fiber. The microbial protein amino acid profile was similar to alfalfa, except for less cysteins. The microbial cells had more thiamine and less niacin than Torula yeast. In vitro digestibility of the microbial protein was 41.2 to 55%, that of cellulose was 52%.


An image processing approach to computing distances between RNA secondary structures dot plots.
Computing the distance between two RNA secondary structures can contribute in understanding the functional relationship between them. When used repeatedly, such a procedure may lead to finding a query RNA structure of interest in a database of structures. Several methods are available for computing distances between RNAs represented as strings or graphs, but none utilize the RNA representation with dot plots. Since dot plots are essentially digital images, there is a clear motivation to devise an algorithm for computing the distance between dot plots based on image processing methods. We have developed a new metric dubbed 'DoPloCompare', which compares two RNA structures. The method is based on comparing dot plot diagrams that represent the secondary structures. When analyzing two diagrams and motivated by image processing, the distance is based on a combination of histogram correlations and a geometrical distance measure. We introduce, describe, and illustrate the procedure by two applications that utilize this metric on RNA sequences. The first application is the RNA design problem, where the goal is to find the nucleotide sequence for a given secondary structure. Examples where our proposed distance measure outperforms others are given. The second application locates peculiar point mutations that induce significant structural alternations relative to the wild type predicted secondary structure. The approach reported in the past to solve this problem was tested on several RNA sequences with known secondary structures to affirm their prediction, as well as on a data set of ribosomal pieces. These pieces were computationally cut from a ribosome for which an experimentally derived secondary structure is available, and on each piece the prediction conveys similarity to the experimental result. Our newly proposed distance measure shows benefit in this problem as well when compared to standard methods used for assessing the distance similarity between two RNA secondary structures. Inspired by image processing and the dot plot representation for RNA secondary structure, we have managed to provide a conceptually new and potentially beneficial metric for comparing two RNA secondary structures. We illustrated our approach on the RNA design problem, as well as on an application that utilizes the distance measure to detect conformational rearranging point mutations in an RNA sequence.


The use of pulmonary artery catheters in intensive care: time for reappraisal?
The precise role of the pulmonary artery catheter (PAC) in reducing the morbidity and mortality of intensive care patients remains uncertain. Future studies of the different patient groups who possibly benefit from their use may well require multicentre trials in order to include sufficient numbers and produce significant conclusions. This would suggest a need for a consensus opinion on how PACs are actually used to obtain the different physiological variables which can influence patient management. A questionnaire was designed to assess the degree of conformity of PAC use in intensive care; the questions were derived from a Medline search and considered by the authors to represent a cross-section of the different aspects and potential errors of PAC use. A postal questionnaire was sent from a District General Hospital to 42 adult Intensive Care Units (ICUs) requesting information on the use of PACs in each ICU. The reply rate was 93%, with all but one of the respondents using PACs. The questionnaire obtained information in different areas of PAC use: 1. General usage of PACs. 2. Measurement of patient height and weight. 3. Confirmation of position of PAC tip. 4. Measurement of pulmonary artery wedge pressure (PAWP). 5. Measurement of cardiac output (CO). 6. Derived values. 7. Monitoring for PAC-related sepsis. The results of the questionnaire demonstrated a considerable lack of standardisation in the use of PACs in the United Kingdom.(ABSTRACT TRUNCATED AT 250 WORDS)


PEGylated liposomes of anastrozole for long-term treatment of breast cancer: in vitro and in vivo evaluation.
The aim of present study was to develop conventional and PEGylated (long circulating), liposomes containing anastrozole (ANS) for effective treatment of breast cancer. ANS is a third-generation non-steroidal aromatase inhibitor of the triazole class used for the treatment of advanced and late-stage breast cancer in post-menopausal women. Under such disease conditions the median duration of therapy should be prolonged until tumor regression ends (>31 months). Liposomes were prepared by the thin film hydration method by using ANS and various lipids such as soyaphosphatidyl choline, cholesterol and methoxy polyethylene glycol distearoyl ethanolamine in different concentration ratios and evaluated for physical characteristics, in vitro drug release and stability. Optimized formulations of liposome were studied for in vitro cytotoxic activity against the BT-549 and MCF-7 cell lines and in vivo behavior in Wistar rats. Preformulation studies, both Fourier transform infrared study and differential scanning calorimetry analysis showed no interaction between the drug and the excipients used in the formulations. The optimized formulations AL-07 and AL-09 liposomes showed encapsulation efficiencies in the range 65.12 ± 1.05% to 69.85 ± 3.2% with desired mean particle size distribution of 101.1 ± 5.9 and 120.2 ± 2.8 nm and zeta potentials of -43.7 ± 4.7 and -62.9 ± 3.5 mV. All the optimized formulations followed Higuchi-matrix release kinetics and when plotted in accordance with the Korsemeyer-Peppas method, the n-value 0.5 < n < 1.0 suggests an anomalous (non-Fickian) transport. Likewise, the PEGylated liposomes showed greater tumor growth inhibition on BT-549 and MCF-7 cell lines from in vitro cytotoxicity studies (p < 0.05). Pharmacokinetic study of conventional and PEGylated liposomes in Wistar rats demonstrated a 3.33- and 20.28-fold increase in AUC(0-∞) values when compared to pure drug (p < 0.001). Among the formulations, PEGylated liposomes showed encouraging results by way of their long circulation and sustained delivery properties for effective treatment of breast cancer.


Quorum size of Pseudomonas syringae is small and dictated by water availability on the leaf surface.
The paradigm of bacterial quorum sensing (QS), which mediates cell-density-dependent gene expression, usually has been studied in high-cell-density planktonic liquid cultures or in biofilms in which signal concentrations accumulate to sufficiently high levels to induce QS. Presumably under conditions with restricted diffusion of the signal molecule, smaller population sizes could achieve such a state of QS induction. The plant-pathogenic bacterium Pseudomonas syringae, in which QS controls traits involved in epiphytic fitness and virulence, occurs on leaf surfaces in aggregates of various sizes. Because leaves often harbor limited surface water, we investigated the size of aggregates that would permit QS in a nonsaturated environment. QS induction was visualized via dual fluorescence of P. syringae cells harboring a transcriptional fusion of mRFP1 with ahlI, which exhibits N-acyl homoserine lactone-dependent transcriptional activity, and a constitutive GFP marker to account for all P. syringae cells on a leaf. Confocal microscopy revealed that, on wet leaves, no QS induction was evident within 2 days after inoculation, but it increased rapidly with increasing aggregate sizes >40 and 22 cells per aggregate by 3 and 4 days, respectively. In contrast, QS induction was common in aggregates >33 cells by 2 days after inoculation on dry leaves and increased rapidly with increasing aggregate sizes >35 and 13 cells after 3 and 4 days, respectively. These observations demonstrate that small groups of cells experience QS conditions on dry leaves where signal diffusion is restricted. Quorum size of bacteria in non-water-saturated environments such as on leaves is small, and QS induction may be commonly operative.


Inhibitory effects of human neutrophil granules and oxygen radicals on adherence of Candida albicans.
The adherence of Candida albicans to dacron fibre microcolumns was significantly suppressed after interaction with human neutrophils. The adherence-inhibiting properties of neutrophils were shown to reside in their cytoplasmic granules and granular enzymes. Oxygen-derived free radicals produced by the respiratory burst may also be responsible, as shown by experiments in which oxygen radicals were generated by the cell-free hypoxanthine-xanthine oxidase system. Dose-response studies with H2O2 and beta-glucoronidase demonstrated that lower concentrations of these agents inhibited adherence without affecting viability of C. albicans. These results suggest that interference with adherence mechanisms may be an effective means of host defence by neutrophils against the colonisation of mucosal surfaces by C. albicans.


The fate of nitrogen in the Zarin-Gol River receiving trout farm effluent.
This study investigated the Zarrin-Gol River ecosystem in Iran to trace organic matter in the food web and evaluate the impact of aquaculture farm effluent using stable isotopes of nitrogen (δ15N) and carbon (δ13C). Using a previously-developed model (Islam 2005), we estimated that a trout farm in the vicinity released 1.4 tons of nitrogen into the river. This was comparable to an estimated total nutrient load of 2.1 tons of nitrogen for the six-month fish-rearing period based on a web-based constituent load estimator (LOADEST). A model estimate of river nitrogen concentration at the time of minimum river discharge (100 L/s) was 2.74 mg/L. Despite relatively high nitrogen loading from the farm, isotope data showed typical food web structure. Several biological groups had elevated δ13C or δ15N values, but there was limited evidence for the entry of organic matter from the trout farm into the food web, with sites above and below trout farms having inconsistent patterns in 15N enrichment. By coupling nitrogen load modeling with stable isotope analysis we showed that stable isotopes might not be effective tracers of organic matter into food webs, depending on surrounding land use and other point sources of nutrients. The Zarrin-Gol River ecosystem, like other basins with high human population density, remains vulnerable to eutrophication in part due to trout farm effluent.


Prokaryotic and eukaryotic community structure in field and cultured microbialites from the alkaline Lake Alchichica (Mexico).
The geomicrobiology of crater lake microbialites remains largely unknown despite their evolutionary interest due to their resemblance to some Archaean analogs in the dominance of in situ carbonate precipitation over accretion. Here, we studied the diversity of archaea, bacteria and protists in microbialites of the alkaline Lake Alchichica from both field samples collected along a depth gradient (0-14 m depth) and long-term-maintained laboratory aquaria. Using small subunit (SSU) rRNA gene libraries and fingerprinting methods, we detected a wide diversity of bacteria and protists contrasting with a minor fraction of archaea. Oxygenic photosynthesizers were dominated by cyanobacteria, green algae and diatoms. Cyanobacterial diversity varied with depth, Oscillatoriales dominating shallow and intermediate microbialites and Pleurocapsales the deepest samples. The early-branching Gloeobacterales represented significant proportions in aquaria microbialites. Anoxygenic photosynthesizers were also diverse, comprising members of Alphaproteobacteria and Chloroflexi. Although photosynthetic microorganisms dominated in biomass, heterotrophic lineages were more diverse. We detected members of up to 21 bacterial phyla or candidate divisions, including lineages possibly involved in microbialite formation, such as sulfate-reducing Deltaproteobacteria but also Firmicutes and very diverse taxa likely able to degrade complex polymeric substances, such as Planctomycetales, Bacteroidetes and Verrucomicrobia. Heterotrophic eukaryotes were dominated by Fungi (including members of the basal Rozellida or Cryptomycota), Choanoflagellida, Nucleariida, Amoebozoa, Alveolata and Stramenopiles. The diversity and relative abundance of many eukaryotic lineages suggest an unforeseen role for protists in microbialite ecology. Many lineages from lake microbialites were successfully maintained in aquaria. Interestingly, the diversity detected in aquarium microbialites was higher than in field samples, possibly due to more stable and favorable laboratory conditions. The maintenance of highly diverse natural microbialites in laboratory aquaria holds promise to study the role of different metabolisms in the formation of these structures under controlled conditions.


Effects of UVB radiation on 4-hydroxy-2-trans-nonenal metabolism and toxicity in human keratinocytes.
Endogenous lipid peroxidation (LPO)-derived aldehydes accumulate in human skin after photoexposure and contribute to the development of skin cytotoxicity and cancer. This study employed LC-ESI-MS and HPLC-UV-DAD techniques to investigate the effect of UVB radiation on the biotransformation and detoxification of the prototype aldehyde 4-hydroxy-2-trans-nonenal (HNE) using the human keratinocyte cell line (NCTC 2544). In parallel we followed the keratinocytes' cytotoxic response to HNE through morphological analysis and cell viability assay. In UVB-unstressed keratinocytes, even a supraphysiological dose of the aldehyde (200 microM) was rapidly and completely cleared in metabolized form (free and GSH-conjugated metabolites) from the cell, with no signs of cytotoxicity. By contrast, UVB preexposure already at 1 MED (50 mJ/cm2, the minimal erythemal dose in humans) markedly impaired HNE metabolism. After 2 h of incubation, the relative amount of GSH-conjugated adducts dose-dependently dropped from 44% (unirradiated cells) to 22% at 3 MED as a consequence of UVB-induced GSH depletion (no impairment of GST A4.4 nor of G6PD activities was observed). The levels of free metabolites, 1,4-dihydroxy-trans-nonene (DHN) and 4-hydroxy-trans-2-nonenoic acid (HNA), were modified (+30% DHN, -22% HNA) only at 3 MED, in parallel to the AR and ALDH enzyme activity modulation. In addition, a dose-dependent increase of unmodified HNE was found in the extracellular medium, paralleled by a significant fraction of the HNE-incubated dose not recovered at the intra- or extracellular level. The impairment of HNE metabolism paralleled a dramatic cytotoxic response. These results provide a reasonable explanation for the massive accumulation of carbonyl toxins in human skin in vivo after photoexposure and shed light on the detrimental effects of UVB radiation in the presence of unmetabolized LPO metabolites.


Trk C receptor signaling regulates cardiac myocyte proliferation during early heart development in vivo.
Neurotrophin-3 (NT-3) is a member of the neurotrophin family of growth factors, best characterized by its survival- and differentiation-inducing effects on developing neurons bearing the trk C receptor tyrosine kinase. Through analysis of NT-3 and trk C gene-targeted mice we have identified NT-3 as critically regulating cardiac septation, valvulogenesis, and conotruncal formation. Although these defects could reflect cardiac neural crest dysfunction, the expression of NT-3 and trk C by cardiac myocytes prior to neural crest migration prompted analysis of cell-autonomous actions of NT-3 on cardiac myocytes. Retroviral-mediated overexpression of truncated trk C receptor lacking kinase activity was used to inhibit activation of trk C by endogenous NT-3, during early heart development in ovo. During the first week of chicken development, expression of truncated trk C reduced myocyte clone size by more than 60% of control clones. Direct mitogenic actions of NT-3 on embryonic cardiac myocytes were demonstrated by analysis of BrdU incorporation or PCNA immunoreactivity in control and truncated trk C-expressing clones. Inhibition of trk C signaling reduced cardiac myocyte proliferation during the first week of development, but had no effect at later times. These studies demonstrate that endogenous NT-3:trk C signaling regulates cardiac myocyte proliferation during cardiac looping and the establishment of ventricular trabeculation but that myocyte proliferation becomes NT-3 independent during the second week of embryogenesis.


Combined system for the treatment and reuse of urban wastewater: the efficiency of anaerobic reactors + hybrid constructed wetlands + ozonation.
The research developed a combined system in batch flow and in pilot scale for the treatment and reuse of urban effluents. The system was fed raw effluent from a university campus in Brazil and composed of four anaerobic reactors, three constructed wetlands (CWs) and an ozonation unit. The three sequential hybrid constructed wetlands were composed of a floating treatment wetland, an aerobic-anoxic baffled constructed wetland (CW) and a saturated vertifcal flow CW. Later, during the last trimester, weekly samples of the treated effluent were ozonated by bubbling with an application rate of 240 mg.h-1 O3. The system presented high removal rates efficiencies in terms of carbonaceous organic matter (78.9%), nitrogen (91.0%), color (96.7%) and turbidity (99.1%). In addition, it worked well for disinfection and acute ecotoxicity, but P was only efficiently (75%) removed in the first 8 months, with removing efficiency declining after this period. Ozonation provided significant color removal and an increased pH. The combination of floating, alternated upflow and downflow and saturated vertical flows improved the treatment of wastewater. This was due to the presence of both aerobic and anaerobic zones, as well as the filter substrate, through an integrated system with simple construction and operation and increased lifespan.


Synergistic effect of TGF-beta superfamily members on the induction of Foxp3+ Treg.
TGF-beta plays an important role in the induction of Treg and maintenance of immunologic tolerance, but whether other members of TGF-beta superfamily act together or independently to achieve this effect is poorly understood. Although others have reported that the bone morphogenetic proteins (BMP) and TGF-beta have similar effects on the development of thymocytes and T cells, in this study, we report that members of the BMP family, BMP-2 and -4, are unable to induce non-regulatory T cells to become Foxp3+ Treg. Neutralization studies with Noggin have revealed that BMP-2/4 and the BMP receptor signaling pathway is not required for TGF-beta to induce naïve CD4+CD25- cells to express Foxp3; however, BMP-2/4 and TGF-beta have a synergistic effect on the induction of Foxp3+ Treg. BMP-2/4 affects non-Smad signaling molecules including phosphorylated ERK and JNK, which could subsequently promote the differentiation of Foxp3+ Treg induced by TGF-beta. Data further advocate that TGF-beta is a key signaling factor for Foxp3+ Treg development. In addition, the synergistic effect of BMP-2/4 and TGF-beta indicates that the simultaneous manipulation of TGF-beta and BMP signaling might have considerable effects in the clinical setting for the enhancement of Treg purity and yield.


CpG domains downstream of TSSs promote high levels of gene expression.
CpG dinucleotides are known to play a crucial role in regulatory domains, affecting gene expression in their natural context. Here, we demonstrate that intragenic CpG frequency and distribution impacts transgene and genomic gene expression levels in mammalian cells. As shown for the Macrophage Inflammatory Protein 1α, de novo RNA synthesis correlates with the number of CpG dinucleotides, whereas RNA splicing, stability, nuclear export and translation are not affected by the sequence modification. Differences in chromatin accessibility in vivo and altered nucleosome positioning in vitro suggest that increased CpG levels destabilize the chromatin structure. Moreover, enriched CpG levels correlate with increased RNA polymerase II elongation rates in vivo. Interestingly, elevated CpG levels particularly at the 5' end of the gene promote efficient transcription. We show that this is a genome-wide feature of highly expressed genes, by identifying a domain of ∼700 bp with high CpG content downstream of the transcription start site, correlating with high levels of transcription. We suggest that these 5' CpG domains are required to distort the chromatin structure and to increase gene activity.


[Lateralization of visual perception of letters during reverse masking].
Identification by healthy subjects of single letters in conditions of backward masking was studied at lateralized presentation of stimuli. Both at dichoptic and unilateral presentations, the letters are identified better if they are presented in the left visual field, i.e. addressed "directly" to the right hemisphere. It is suggested that the data obtained result from the specificity of hemispheres' cooperation under difficult conditions of visual-spatial stage of information processing.


BEP relations for N2 dissociation over stepped transition metal and alloy surfaces.
We present density functional theory (DFT) calculations for N2 dissociation on stepped face-centred cubic (211) surface slabs. By using the same crystal structure, the same adsorption site for atomic nitrogen, and the same transition-state bond length of N2 over a range of pure metal surfaces, a perfectly linear Brønsted-Evans-Polanyi (BEP) relation between the transition-state potential energy and the dissociative chemisorption energy is obtained. The perfect BEP relation, which extends over 12 eV in chemisorption energy, suggests that the manifestation of BEP relations for surface reactions is a general electronic structure effect, and that geometric effects are responsible for the scatter which is normally observed around the BEP line. The BEP relation is also shown to be valid for both surface and bulk alloys. The scatter is, however, larger than for the pure elements. This can be understood as a larger geometrical variance. To analyze the accuracy of the DFT calculations a detailed convergence study is performed for several adsorbates on stepped hexagonal close-packed and face-centred cubic Ru slabs.


The cestode order Rhinebothriidea no longer family-less: A molecular phylogenetic investigation with erection of two new families and description of eight new species of Anthocephalum.
The spiral intestines of a total of 30 specimens of 14 species of batoids from around the world were examined for rhinebothriideans. These consisted of Taeniura grabata, Dasyatis margaritella, and Dasyatis sp. from Senegal, Dasyatis americana from Florida, Dasyatis dipterura and Dasyatis longa from México, Himantura jenkinsii, Himantura leoparda, Himantura uarnak 2, Urogymnus asperrimus 1, and Neotrygon kuhlii 4 from Australia, in addition to Himantura uarnacoides and Neotrygon kuhlii 1 from Borneo. Each of these hosted one or more species of Anthocephalum. Eleven of the cestode species were new to science; four represented described species. In addition, Urotrygon aspidura from Costa Rica hosted a species of Escherbothrium. Sufficient material was available for formal description of the following eight species of Anthocephalum: A. decrisantisorum n. sp., A. healyae n. sp., A. jensenae n. sp., A. mattisi n. sp., A. meadowsi n. sp., A. odonnellae n. sp., A. papefayi n. sp., and A. philruschi n. sp. These species differ from their nine described congeners in overall size, number of proglottids and marginal loculi, number and arrangement of testes, apical sucker size, arrangement and distribution of vitelline follicles particularly with respect in the post-poral field, and muscularity of the genital pore. The diagnosis of Anthocephalum is emended slightly to accommodate these new species. Material of four previously described Anthocephalum species, seven of the novel Anthocephalum species described here, 3 undescribed Anthocephalum species, and the species of Escherbothrium was preserved in 95% ethanol and partial 28S rDNA (D1-D3) and complete 18S rDNA sequence data were generated de novo. These data were combined with data from GenBank for Anthocephalum cf. centrurum (recognized as A. mattisi n. sp. below) and 29 species representing 12 other putative rhinebothriidean genera. Phylogenetic analyses using Bayesian Inference and Maximum Likelihood methods were conducted using a total of five representatives of the Lecanicephalidea, Cathetocephalidea and "Tetraphyllidea" as outgroups. The analyses yielded trees that were largely congruent and that supported the existence of four major subgroups of rhinebothriideans. Family designations were established for each of these clades. Echeneibothriidae was elevated from subfamily to family level to accommodate the group consisting of Echeneibothrium and Pseudanthobothrium; this family is unique in retaining the apical organ (as a myzorhynchus) into adulthood. Rhinebothriidae was elevated from subfamily to family level to accommodate the group consisting of Rhabdotobothrium, Rhinebothrium, Rhinebothroides, Rhodobothrium, Scalithrium and Spongiobothrium. This family is distinctive in its lack of apical suckers and also of a definitive anterior/posterior orientation to the bothridia. Anthocephaliidae n. fam. was established to house Anthocephalum and taxa identified as New Genus 1, New Genus 2, and New Genus 4 by previous authors. The bothridia of its members exhibit a conspicuous anterior/posterior orientation signaled by the presence of an apical sucker. In addition, its members bear marginal loculi or one or more rows of facial loculi and vitelline follicles that are usually interrupted by the ovary. Escherbothriidae n. fam. was established to house Escherbothrium and the taxon identified as New Genus 3 by previous authors. It most closely resembles Anthocephaliidae but the facial loculi are arranged in columns anteriorly and rows posteriorly, rather than arranged in multiple rows or entirely lacking. A key to the families is also provided.


MiR-301a-3p Advances IRAK1-Mediated Differentiation of Th17 Cells to Promote the Progression of Systemic Lupus Erythematosus via Targeting PELI1.
Systemic lupus erythematosus (SLE) is a common autoimmune disease with high incidence in females. The pathogenesis of SLE is complex, and healing SLE has become a serious challenge for clinical treatment. Aberrant expression of miR-301a-3p involves the progressions of multiple diseases, and some studies have indicated that increased miR-301a-3p could induce the inflammatory injury of some organs. However, the role and molecular mechanism of miR-301a-3p in SLE remain unclear. In this study, the miR-301a-3p levels in peripheral blood mononuclear cells (PBMCs) of the patients with SLE and health subjects were measured with qRT-PCR. The ELISA assay was used to investigate the effect of miR-301a-3p on the levels of inflammatory factors in PBMCs, and flow cytometry assays were used to observe the effect of miR-301a-3p on the levels of CD4+ T cells and Th17 cells in PBMCs. Moreover, TargetScan, dual-luciferase reporter assay, and western blot were used to reveal the downstream targets and regulation mechanism of miR-301a-3p in SLE. The results showed that miR-301a-3p was significantly upregulated in PBMCs of the SLE patients, and increased miR-301a-3p could boost the expression of IL-6, IL-17, and INF-γ in PBMCs and promote the differentiation of Th17 cells. It was found that PELI1 was a target of miR-301a-3p, and PELI1 upregulation could effectively reverse the effect of miR-301a-3p on PBMCs. Besides, this study also found that miR-301a-3p could promote the expression of IRAK1 to involve the progression of SLE via targeting PELI1. In conclusion, this study suggests that increased miR-301a-3p serves as a pathogenic factor in SLE to promote IRAK1-mediated differentiation of Th17 cells via targeting PELI1.


Functional expression of the multimodal extracellular calcium-sensing receptor in pulmonary neuroendocrine cells.
The Ca(2+)-sensing receptor (CaSR) is the master regulator of whole-body extracellular free ionized [Ca(2+)]o. In addition to sensing [Ca(2+)]o, CaSR integrates inputs from a variety of different physiological stimuli. The CaSR is also expressed in many regions outside the [Ca(2+)]o homeostatic system, including the fetal lung where it plays a crucial role in lung development. Here, we show that neuroepithelial bodies (NEBs) of the postnatal mouse lung express a functional CaSR. NEBs are densely innervated groups of neuroendocrine epithelial cells in the lung representing complex sensory receptors in the airways and exhibiting stem cell characteristics. qRT-PCR performed on laser microdissected samples from GAD67-GFP mouse lung cryosections revealed exclusive expression of the CaSR in the NEB microenvironment. CaSR immunoreactivity was present at NEB cells from postnatal day 14 onwards. Confocal imaging of lung slices revealed that NEB cells responded to an increase of [Ca(2+)]o with a rise in intracellular Ca(2+) ([Ca(2+)]i); an effect mimicked by several membrane-impermeant CaSR agonists (e.g. the calcimimetic R-568) and that was blocked by the calcilytic Calhex-231. Block of TRPC channels attenuated the CaSR-dependent increases in [Ca(2+)]i, suggesting that Ca(2+) influx through TRPC channels contributes to the total [Ca(2+)]i signal evoked by the CaSR in NEBs. CaSR also regulated baseline [Ca(2+)]i in NEBs and, through paracrine signaling from Clara-like cells, coordinated intercellular communication in the NEB microenvironment. These data suggest that the NEB CaSR integrates multiple signals converging on this complex chemosensory unit, and is a key regulator of this intrapulmonary airway stem cell niche.


Imaging InlC secretion to investigate cellular infection by the bacterial pathogen Listeria monocytogenes.
Bacterial intracellular pathogens can be conceived as molecular tools to dissect cellular signaling cascades due to their capacity to exquisitely manipulate and subvert cell functions which are required for the infection of host target tissues. Among these bacterial pathogens, Listeria monocytogenes is a Gram positive microorganism that has been used as a paradigm for intracellular parasitism in the characterization of cellular immune responses, and which has played instrumental roles in the discovery of molecular pathways controlling cytoskeletal and membrane trafficking dynamics. In this article, we describe a robust microscopical assay for the detection of late cellular infection stages of L. monocytogenes based on the fluorescent labeling of InlC, a secreted bacterial protein which accumulates in the cytoplasm of infected cells; this assay can be coupled to automated high-throughput small interfering RNA screens in order to characterize cellular signaling pathways involved in the up- or down-regulation of infection.


The effect of parotid gland-sparing intensity-modulated radiotherapy on salivary composition, flow rate and xerostomia measures.
To describe parotid gland (PG) saliva organic and inorganic composition and flow rate changes, after curative intensity-modulated radiotherapy (IMRT) for head and neck cancer (HNC), and analyse the relationship between PG saliva analytes and xerostomia measures. Twenty-six patients recruited to five prospective phase 2 or 3 trials which assessed toxicity and efficacy of IMRT by HNC subsite, provided longitudinal PG saliva. Salivary flow rate, and subjective and objective xerostomia measures were prospectively collected and saliva tested for inorganic and organic analytes. Statistical comparisons of longitudinal analyte changes and analysis for a relationship between dichotomized xerostomia score and saliva analytes were performed. One hundred and forty-two PG saliva samples from 26 patients were analysed. At 3-6 months after IMRT, stimulated and unstimulated saliva showed significantly decreased flow rate, total protein (TP) secretion rate, phosphate concentration and increased lactoferrin (LF) concentration. Stimulated saliva alone had elevated LF secretion rate and beta-2-microglobulin (B2 M) concentration with decreased calcium (Ca2+ ) and magnesium (Mg2+ ) concentrations and Ca2+ secretion rate. At >12 months, under stimulated and unstimulated conditions, increased LF concentration and decreased Mg2+ and phosphate concentration persisted and, in stimulated saliva, there was decreased potassium (K+ ) and Mg2+ concentration. Unstimulated TP secretion rate was lower in the presence of high-grade xerostomia. Otherwise, no relationship between xerostomia grade and PG salivary flow rate, TP and Ca2+ secretion rate was found. Fewer significant differences in PG saliva analytes >12 months after IMRT indicate good functional recovery. Residual xerostomia after IMRT will only be further reduced by addressing the sparing of subsites of the PG or other salivary gland tissues, in addition to the PG.


We are not all the same: The importance of perceived difference in racial ideology and Afrocentricity for African American relationships.
A large body of existing research on African American relationships perpetuates a deficit model that assumes Eurocentric norms and emphasizes between-group differences (e.g., cross-racial comparisons with the majority group-European Americans). The current study examined within-group variability and the influence of culturally unique factors, Afrocentricity, racial ideology, and perceived discrepancy between self and partner on African American relationship processes. Data were collected from 137 self-identified African American adults in same-race, cross-gender relationships. Consistent with the literature on protective values of Afrocentricity, there was an association between reported relationship quality and high levels of one's own and perceived partner's Afrocentricity. Discrepancies between self and partner Afrocentricity were not associated with relationship processes, but higher perceived discrepancies across all four subscales of racial ideology were associated with lower relationship dedication. Higher perceived discrepancies on the humanist and assimilationist subscales were also related to higher levels of conflict. These findings have important clinical implications and demonstrate a need for further research into the nuances of individual factors and dyadic processes that are unique to African American couples.


Role of exonuclease III in the base excision repair of uracil-containing DNA.
Mutants of Escherichia coli K-12 deficient in both exonuclease III (the product of the xth gene) and deoxyuridine triphosphatase (the dut gene product) are inviable at high temperatures and undergo filamentation when grown at such temperatures. In dut mutants, the dUTP pool is known to be greatly enhanced, resulting in an increased substitution of uracil for thymine in DNA during replication. The subsequent removal of uracil from the DNA by uracil-DNA glycosylase produces apyrimidinic sites, at which exonuclease III is known to have an endonucleolytic activity. The lethality of dut xth mutants, therefore, indicates that exonuclease III is important for this base-excision pathway and suggests that unrepaired apyrimidinic sites are lethal. Two confirmatory findings were as follows. (i) dut xth mutants were viable if they also had a mutation in the uracil-DNA glycosylase (ung) gene; such mutants should not remove uracil from DNA and should not, therefore, generate apyrimidinic sites. (ii) In the majority of the temperature-resistant revertants isolated, viability had been restored by a mutation in the dCTP deaminase (dcd) gene; such mutations should decrease dUTP production and hence uracil misincorporation. The results indicate that, in dut mutants, exonuclease III is essential for the repair of uracil-containing DNA and of apyrimidinic sites.


Role of intraoperative sentinel lymph node mapping in the management of carcinoma of the esophagus.
Precise evaluation of lymph node status is one of the most important factors in determining clinical outcome in treating gastro-intestinal (GI) cancer. Sentinel lymph node (SLN) mapping clearly has become highly feasible and accurate in staging GI cancer. This study aims to investigate the feasibility and accuracy of detection of SLN using methylene blue dye in patients with carcinoma of the esophagus and assess its potential role in determining the rational extent of lymphadenectomy in esophageal cancer surgery. Thirty-two patients of esophageal cancer diagnosed on endoscopic biopsy were enrolled in this prospective study. After laparotomy, patent methylene blue was injected into the subserosal layer adjacent to the tumor. SLNs were defined as blue stained nodes within a period of 5 min. Standard radical esophagogastrectomy with lymphadenectomy was performed in all the patients. All the resected nodes were examined postoperatively by routine hematoxylin and eosin stain for elucidating the presence of metastasis, and the negative SLNs were examined further with cytokeratin immunohistochemical staining. SLNs were detected in 26 (81.25%) patients out of 32 patients who were studied. The number of SLNs ranged from 1 to 4 with a mean value of 1.7 per case. The SLNs of esophageal cancer were only found in N1 area in 21 (80.77%) cases, and in N2 or N3 area in only 19.33%. The overall accuracy of the procedure was 75% in predicting nodal metastasis. SLN had a sensitivity of 85.71% in mid esophageal tumors and 93.33% in lower esophageal tumors. The SLN biopsy had sensitivity of 87.5% in the case of squamous cell carcinoma and 92.86% in the cases of adenocarcinoma of the esophagus. The accuracy of the procedure for squamous cell carcinoma and adenocarcinoma was 60% and 76.47%, respectively. SLN mapping is an accurate diagnostic procedure for detecting lymph node metastasis in patients with esophageal cancer and may indicate rational extent of lymphadenectomy in these patients. SLN mapping provides "right nodes" to the pathologists for detailed analysis and appropriate staging, thereby helping in individualizing the multi-modal treatment for esophageal cancer.


The family's adaptation process to their child's hospitalization in an Intensive Care Unit.
To learn about the adaptation process of family members when experiencing their child's hospitalization in an Intensive Care Unit. A qualitative, descriptive and exploratory study conducted at the Pediatrics Unit with family members of children from the Pediatric Intensive Care Unit. The thematic analysis technique was used. Thirteen (13) family members, 12 mothers and one father participated. The following categories emerged from the results: Family impact on children's hospitalization in a Pediatric Intensive Care Unit and Mechanisms for adapting family members to hospitalization in a Pediatric Intensive Care Unit. The mechanisms adopted by the family in the process of adapting the child to admission to the Pediatric Intensive Care Unit were: positive thinking, understanding the treatment and the operation of the unit. Health professionals can offer strategies to the family in order to make hospitalization less traumatic, making it possible to share experiences and expand the knowledge of those involved.


High-Frequency Regeneration of the Drought-Tolerant Tree Melia volkensii Gurke Using Low-Cost Agrochemical Thidiazuron.
Melia volkensii Gurke is a drought-tolerant tree native to East Africa's arid and semiarid lands (ASALs), with vast but underutilized potential for agroforestry and sustainable livelihoods in the ASALs. Its cultivation is limited by difficulties in propagation via conventional means. Full exploitation of the ability of thidiazuron (TDZ) to elicit regeneration in plant tissue cultures, as sole plant growth regulator (PGR), is hampered by high costs. This study tested the effectiveness of a low-cost agrochemical TDZ for in vitro propagation of M. volkensii. Zygotic embryos from mature seeds were cultured on Gamborg's B5 medium containing 0 to 4 mg/L of agrochemical TDZ from Kingtai Chemicals Co.,Ltd., China. Callus induction frequency was 96.67 to 100%. Significantly large callus fresh mass was produced at 0.05 mg/L TDZ concentration (ANOVA, P < 0.001). The effect of TDZ on embryogenicity was significant over certain ranges of concentrations (Anova, P < 0.001). Multiple somatic embryos developed within 14 days of subculture to hormone-free B5 medium. Somatic embryos developed into microshoots which elongated when transferred to 1/2 MS medium supplemented with 0.1 mg/L 6-benzylaminopurine plus 10% coconut water. The Kingtai-TDZ showed a high potency and suitability for use in M. volkensii tissue culture.


Historical perspective: Arthur Kornberg, a giant of 20th century biochemistry.
For physics, the period from the beginning to the middle of the 20th century was one of great scientific excitement and revolutionary discovery. The analogous era for biochemistry, and its offspring, molecular biology, was the second half of the 20th century. One of the most important and influential leaders of this scientific revolution was Arthur Kornberg. The DNA polymerase, which he discovered in 1955 and showed to have the remarkable capacity to catalyze the template-directed synthesis of DNA, contributed in major ways to the present-day understanding of how DNA is replicated and repaired, and how it is transcribed. The discovery of DNA polymerase also permitted the development of PCR and DNA sequencing, upon which much of modern biotechnology is based. Kornberg's studies of DNA replication, which spanned a period of nearly 30 years, culminated in a detailed biochemical description of the mechanism by which a chromosome is replicated. The final years of Kornberg's life were devoted to the study of polyphosphate, which he was convinced had a crucial role in cellular function.


Utilization management, case management, and you.
Historically, most monitoring functions have been carried out by insurance companies. Monitoring costs was considered their fiduciary obligation to their customers. The exercise of this fiduciary obligation kept premiums low, while increasing or maintaining the benefit levels. Risk (the assumption of losses generated by services costing more than the income received from premiums) was assumed by the insurance company and eventually passed to the customer or the payer. Today, risk is being transferred more and more to the provider. This transfer was started by the creation of DRGs, the main purpose of which was to transfer risk from payers (insurance companies, employers, state and federal government, etc.) to provider health care organizations (physician groups, individual practitioners, hospitals, clinics, etc.).


Renal function and histology after acute hemorrhage in rats under dexmedetomidine action.
About 50 % of indications for dialysis in acute renal failure are related to problems originated during the perioperative period. Intraoperative hemodynamic changes lead to renal vasoconstriction and hypoperfusion. Previous studies have not defined the dexmedetomidine renal role in hemorrhage situations. This study evaluated the effect of dexmedetomidine on renal function and histology after acute hemorrhage in rats. Covered study with 20 Wistars rats, anesthetized with sodium pentobarbital, 50 mg.kg(-1), intraperitoneal, randomized into 2 groups submitted to 30% volemia bleeding: DG - iv dexmedetomidine, 3 microg.kg(-1) (10 min) and continuous infusion - 3 microg.kg(-1).h(-1); CG - pentobarbital. For renal clearance estimative, sodium p-aminohippurate and iothalamate were administered. Studied attributes: heart rate, mean arterial pressure, rectal temperature, hematocrit, iothalamate and p-aminohippurate clearance, filtration fraction, renal blood flow, renal vascular resistance, and histological evaluations of the kidneys. DG showed smaller values of heart rate, mean arterial pressure, and renal vascular resistance, but iothalamate clearance and filtration fraction values were higher. There was similarity in p-aminohippurate clearance and renal blood flow. Both groups had histological changes ischemia-like, but dexmedetomidine determined higher tubular dilatation scores. In rats, after acute hemorrhage, dexmedetomidine determined better renal function, but higher tubular dilation scores.


Clinical features of children with Henoch-Schonlein purpura: risk factors associated with renal involvement.
This study aimed to evaluate renal involvement and factors affecting the prognosis in patients with Henoch-Schonlein purpura (HSP). The outcomes of 107 children diagnosed with HSP who had been followed up for at least 6 months were reviewed. Renal involvement was observed in 26.1% of the patients. The mean age of the patients with renal involvement was 8.8 ± 4.0 years as compared to 7.1 ± 2.9 years in the patients without renal involvement (P = .02). The risk of renal involvement was found to be significantly higher in the patients who were 10 years old and over (P < .001). In the group with renal involvement, the frequency of scrotal involvement was significantly higher than that of the group without renal involvement (P = .02). The mean serum immunoglobulin A level of the patients with renal involvements was significantly higher (P = .04) and the mean serum complement C3 levels was significantly lower (P = .04) than those of the patients without renal involvement. None of the patients with renal involvement reached end-stage kidney failure. No significant relationship was observed between the development of renal involvement and early steroid treatment. This study proposes that in old children with HSP, elevated serum immunoglobulin A levels, decreased serum complement C3 levels, and scrotal involvement are associated with renal involvement. We failed to find any effect of steroid treatment on development of renal involvement.


Norwalk virus: how infectious is it?
Noroviruses are major agents of viral gastroenteritis worldwide. The infectivity of Norwalk virus, the prototype norovirus, has been studied in susceptible human volunteers. A new variant of the hit theory model of microbial infection was developed to estimate the variation in Norwalk virus infectivity, as well as the degree of virus aggregation, consistent with independent (electron microscopic) observations. Explicit modeling of viral aggregation allows us to express virus infectivity per single infectious unit (particle). Comparison of a primary and a secondary inoculum showed that passage through a human host does not change Norwalk virus infectivity. We estimate the average probability of infection for a single Norwalk virus particle to be close to 0.5, exceeding that reported for any other virus studied to date. Infected subjects had a dose-dependent probability of becoming ill, ranging from 0.1 (at a dose of 10(3) NV genomes) to 0.7 (at 10(8) virus genomes). A norovirus dose response model is important for understanding its transmission and essential for development of a quantitative risk model. Norwalk virus is a valuable model system to study virulence because genetic factors are known for both complete and partial protection; the latter can be quantitatively described as heterogeneity in dose response models.


Singlet Oxygen Generation via UV-A, -B, and -C Photoexcitation of Isoprene-Oxygen (C5H8-O2) Encounter Complexes in the Gas Phase.
The formation of singlet oxygen 1O2 provided by the photoexcitation of the encounter complexes of isoprene with oxygen (C5H8-O2) in the gas phase within the spectral region 253.5-355 nm has been observed at the elevated pressure of oxygen. Singlet oxygen has been detected with its NIR luminescence centered near 1.27 μm. The photogeneration of 1O2 is found to be a one-photon process. In the UV-C region (253-278 nm) the quantum yield of 1O2 is measured. This yield of 1O2 is governed mainly by photoexcitation of O2 molecules to the Herzberg III (3Δu) state via enhanced absorption by C5H8-O2 collision complexes. So excited triplet O2 gives rise to singlet oxygen because of triplet-triplet annihilation in the collisions with unexcited O2 molecules. In the UV-B (308 nm) region the appearance of 1O2 is attributed to the excitation of a double spin-flip (DSF) transition in complex C5H8-O2. In the UV-A region (355 nm) besides DSF the O2-assisted T1 ← S0 excitation of isoprene to the triplet state takes place, which is a sensitizer of 1O2 formation. The contribution of the encounter complexes C5H8-O2 to the production of singlet oxygen and to the lifetime of isoprene in the Earth's troposphere are estimated.


Unusual presentation of giant cell angiofibroma of the eyelids.
To describe a case of bilateral eyelid-confined giant cell angiofibroma (GCAF) in a patient with a slowly progressive bilateral eyelid swelling. A 40-year-old man with a 5-year history of slowly progressive bilateral eyelid swelling, severe functional impairment, and bilateral cosmetic deformity was studied. An extensive ophthalmologic evaluation, laboratory examinations, and orbital magnetic resonance imaging were carried out. Clinical examination showed nonpitting lymphedema affecting both upper and lower eyelids, with orange peel skin. Orbital magnetic resonance imaging revealed diffuse thickening of the preseptal structures in the eyelids without extension to the orbit. Histologic specimen revealed the presence of spindle and multinucleated giant cells surrounding pseudovascular spaces strongly positive to CD34 and vimentin. A diagnosis of GCAF was made and radiation therapy was performed 3 weeks after surgical debulking with partial recovery of visual and anatomic function. Giant cell angiofibroma involving the eyelid is rare and can represent a diagnostic and therapeutic challenge to the ophthalmologist.


Natural (-)- and unnatural (+)-enantiomers of morphine: comparative metabolism and effect of morphine and phenobarbital treatment.
The comparative metabolism of the natural (-)- and the unnatural (+)-morphine was studied in liver microsomes from phenobarbital-treated, morphine-treated and control rats. (-)-Morphine was glucuronidated only at position 3 with the formation of (-)-morphine-3-glucuronide [(-)-M3G] at an average rate of 2.19 nmol X mg protein-1 X min-1. In contrast, (+)-morphine was conjugated preferentially at position 6. The rate of formation of (+)-morphine-6-glucuronide [(+)-M6G] was 3.1 times higher than that of (+)-M3G (0.92 and 0.30 nmol X mg protein-1 X min-1, respectively). Natural morphine was N-demethylated at an average rate of 0.42 nmol X mg protein-1 X min-1, and this rate of N-demethylation was about twice as high as that of (+)-morphine (0.17 nmol X mg protein-1 X min-1). Phenobarbital treatment led to a 3- to 4-fold increase in the formation rate of (-)-M3G and (+)-M6G whereas the glucuronidation of (+)-morphine at position 3 was increased only marginally. No change in the N-demethylation of either enantiomer was observed. In contrast, pretreatment with (-)-morphine decreased the N-demethylation of both enantiomers by about 80% without any effect on the glucuronidation. The increased ratio of the rate of (+)-M6G/(+)-M3G formation after phenobarbital treatment indicates that different isozymes may be involved in the two reactions. This study demonstrates that there is an inherent substrate stereoselectivity and site selectivity for morphine in the rat hepatic uridine 5'-diphosphate-glucuronyltransferase although not as critical as for the opiate receptors mediating analgesia.


Association between sugar-sweetened and artificially sweetened soft drinks and type 2 diabetes: systematic review and dose-response meta-analysis of prospective studies.
The intake of sugar-sweetened soft drinks has been reported to be associated with an increased risk of type 2 diabetes, but it is unclear whether this is because of the sugar content or related lifestyle factors, whether similar associations hold for artificially sweetened soft drinks, and how these associations are related to BMI. We aimed to conduct a systematic literature review and dose-response meta-analysis of evidence from prospective cohorts to explore these issues. We searched multiple sources for prospective studies on sugar-sweetened and artificially sweetened soft drinks in relation to the risk of type 2 diabetes. Data were extracted from eleven publications on nine cohorts. Consumption values were converted to ml/d, permitting the exploration of linear and non-linear dose-response trends. Summary relative risks (RR) were estimated using a random-effects meta-analysis. The summary RR for sugar-sweetened and artificially sweetened soft drinks were 1·20/330 ml per d (95 % CI 1·12, 1·29, P< 0·001) and 1·13/330 ml per d (95 % CI 1·02, 1·25, P= 0·02), respectively. The association with sugar-sweetened soft drinks was slightly lower in studies adjusting for BMI, consistent with BMI being involved in the causal pathway. There was no evidence of effect modification, though both these comparisons lacked power. Overall between-study heterogeneity was high. The included studies were observational, so their results should be interpreted cautiously, but findings indicate a positive association between sugar-sweetened soft drink intake and type 2 diabetes risk, attenuated by adjustment for BMI. The trend was less consistent for artificially sweetened soft drinks. This may indicate an alternative explanation, such as lifestyle factors or reverse causality. Future research should focus on the temporal nature of the association and whether BMI modifies or mediates the association.


Approximate bayesian computation without summary statistics: the case of admixture.
In recent years approximate Bayesian computation (ABC) methods have become popular in population genetics as an alternative to full-likelihood methods to make inferences under complex demographic models. Most ABC methods rely on the choice of a set of summary statistics to extract information from the data. In this article we tested the use of the full allelic distribution directly in an ABC framework. Although the ABC techniques are becoming more widely used, there is still uncertainty over how they perform in comparison with full-likelihood methods. We thus conducted a simulation study and provide a detailed examination of ABC in comparison with full likelihood in the case of a model of admixture. This model assumes that two parental populations mixed at a certain time in the past, creating a hybrid population, and that the three populations then evolve under pure drift. Several aspects of ABC methodology were investigated, such as the effect of the distance metric chosen to measure the similarity between simulated and observed data sets. Results show that in general ABC provides good approximations to the posterior distributions obtained with the full-likelihood method. This suggests that it is possible to apply ABC using allele frequencies to make inferences in cases where it is difficult to select a set of suitable summary statistics and when the complexity of the model or the size of the data set makes it computationally prohibitive to use full-likelihood methods.


Regression of myocardial hypertrophy and influence of adrenergic system.
Studies of regression of myocardial hypertrophy in spontaneously hypertensive rats (SHR) suggest that the adrenergic system may play an important role in the reversal of hypertrophy. The effect of propranolol on reversal of hypertrophy, however, is still controversial. This study describes the effect of propranolol, given alone or in combination with hydralazine in different ratios for 4 wk, on blood pressure (BP), ventricular weight, and myocardial catecholamine (MC) concentrations. The data show that a certain ratio of propranolol to hydralazine (750:30) leads to moderate BP control (196-156 mmHg) without increased MC (634 vs. 552 ng/g) and moderately reduced hypertrophy. Reduction of BP alone with increased MC (hydralazine alone) or reduction of MC without BP control (propranolol alone) failed to reduce hypertrophy. A significant correlation between both ventricular weight and heart rate with MC (r = 0.6) was obtained by multiple regression analysis. This study suggests that adrenergic factors seem to play an important role in modulating structural cardiac response to variations in arterial pressure.


Insulin responsiveness in non-fa/fa and fa/fa Zucker rats: effects of adipocyte size.
Epididymal adipose tissue from non-fa/fa (lean) and fa/fa (obese) 14.5-week-old Zucker rats was used to study the influence of insulin and genotype on uptake of glucose and palmitate into adipocytes of different sizes. After incubation with radioactive substrate, adipocytes were inactivated and fixed by addition of osmium tetroxide; fixed adipocytes were isolated and separated by screening on the basis of size. Rates of substrate uptake into triacylglycerols were measured in adipocytes of each of ten size categories. Uptake rates of both glucose and palmitate increased as adipocyte size increased. Insulin had no effect on glucose uptake per adipocyte for fa/fa rats but had a highly significant (P less than 0.01) stimulatory effect on that for non-fa/fa rats. This stimulation became significantly greater with increasing adipocyte size. When insulin was included in the incubation media, glucose uptake rates were similar between similar sizes of adipocytes from non-fa/fa and fa/fa rats. Absence of insulin from the incubation media, however, resulted in lower rates of glucose uptake by adipocytes from non-fa/fa rats. Glucose uptake was maximal in adipocytes from fa/fa rats, even in the absence of insulin. Net uptake of palmitate into triacylglycerols was not influenced by insulin; a significant interaction was observed, however, between adipocyte size and genotype. Large adipocytes from fa/fa rats had greater rates of palmitate uptake than did adipocytes of similar size from non-fa/fa rats. The reverse was true for adipocytes less than 125 micron in diameter. The results of this study show that response to insulin of adipocytes of difference sizes varies with adipocyte size and with genotype.


Precision measurement of the fundamental vibrational frequencies of tritium-bearing hydrogen molecules: T2, DT, HT.
High-resolution coherent Raman spectroscopic measurements of all three tritium-containing molecular hydrogen isotopologues T2, DT and HT were performed to determine the ground electronic state fundamental Q-branch (v = 0 → 1, ΔJ = 0) transition frequencies at accuracies of 0.0005 cm-1. An over hundred-fold improvement in accuracy over previous experiments allows the comparison with the latest ab initio calculations in the framework of non-adiabatic perturbation theory including nonrelativisitic, relativisitic and QED contributions. Excellent agreement is found between experiment and theory, thus providing a verification of the validity of the NAPT-framework for these tritiated species. While the transition frequencies were corrected for ac-Stark shifts, the contributions of non-resonant background as well as quantum interference effects between resonant features in the nonlinear spectroscopy were quantitatively investigated, also leading to corrections to the transition frequencies. Methods of saturated CARS with the observation of Lamb dips, as well as the use of continuous-wave radiation for the Stokes frequency were explored, that might pave the way for future higher-accuracy CARS measurements.


[PLXNA1 is highly expressed in hepatocellular carcinoma and affects patients' survival and immune microenvironment].
To investigate PLXNA1 expression in hepatocellular carcinoma (HCC) and explore its biological function and impacts on patients' survival outcomes and immune microenvironment. Bioinformatic analysis of highly expressed immune-related genes in HCC were performed using TCGA database and Immport website, and 7 genes associated with the survival outcomes of the patients were identified using univariate Cox regression analysis, Gene Expression Profiling Interactive Analysis, and Kaplan Meier plotter website. The expression profile of PLXNA1 in HCC was verified using GEO database. The impact of PLXNA1 expression on survival outcomes of HCC patients was analyzed using TCGA database, Kaplan Meier, and timeROC curve analyses, and its association with immune cell infiltration was explored using TIMER website, CIBERSORT, and ssGSEA. Immunohistochemmistry was used to detect PLXNA1 expression in clinical specimens of HCC and adjacent tissues, and the correlation of PLXNA1 expression level with the patients' survival was analyzed. RT-qPCR was used to examine PLXNA1 expressions in different HCC cell lines, and the effects of PLXNA1 knockdown on proliferation and migration of SMMC-7721 cells were evaluated using CCK-8 and Transwell assays. Bioinformatic analyses suggested that PLXNA1 was highly expressed in HCC, and its high expression was associated with poor survival outcomes of the patients. PLXNA1 expression level was significantly correlated with immune cell infiltration in HCC. Immunohistochemmistry showed that compared with the adjacent tissues, HCC tissues had significantly higher PLXNA1 expressions, which were associated with a poor patient survival and served also as a diagnostic indicator for HCC (AUC= 0.9346). In cultured HCC cell lines, SMMC-7721 cells showed a higher PLXNA1 expression than HL-7702 cells, and PLXNA1 knockdown significantly suppressed proliferation and migration of SMMC-7721 cells. PLXNA1 is highly expressed in HCC to promote tumor cell migration and proliferation and affect the patients' survival outcomes and immune microenvironment.


"I Wasn't Presented With Options": Perspectives of Black Veterans Receiving Care for Uterine Fibroids in the Veterans Health Administration.
Black women with uterine fibroids experience greater symptom severity and worse treatment outcomes compared with their White counterparts. Black veterans who use Veterans Health Administration (VA) health care experience similar disparities. This study investigated the experiences of Black veterans receiving care for uterine fibroids at VA. We identified Black veterans aged 18 to 54 years with newly diagnosed symptomatic uterine fibroids between the fiscal years 2010 and 2012 using VA medical record data, and we recruited participants for interviews in 2021. We used purposive sampling by the last recorded fibroid treatment in the data (categorized as hysterectomy, other uterine-sparing treatments, and medication only/no treatment) to ensure diversity of treatment experiences. In-depth semistructured interviews were conducted to gather rich narratives of veterans' uterine fibroid care experiences. Transcribed interviews were analyzed using content analysis. Twenty Black veterans completed interviews. Key themes that emerged included the amplified impact of severe fibroid symptoms in male-dominated military culture; the presence of multilevel barriers, from individual to health care system factors, that delayed access to high-quality treatment; insufficient treatments offered; experiences of interpersonal racism and provider bias; and the impact of fertility loss related to fibroids on mental health and intimate relationships. Veterans with positive experiences stressed the importance of finding a trustworthy provider and self-advocacy. System-level interventions, such as race-conscious and person-centered care training, are needed to improve care experiences and outcomes of Black veterans with fibroids.


Increased expression of OsSPX1 enhances cold/subfreezing tolerance in tobacco and Arabidopsis thaliana.
Low temperature is a major environmental stress for plants. Many important cultivated crops have limited capacity to survive below freezing/subfreezing temperatures. Low inorganic phosphate (Pi) is reportedly important in triggering cold acclimatization. SPX (SYG1/Pho81/XPR1: SYG1, suppressor of yeast gpal; Pho81, CDK inhibitor in yeast PHO pathway; XPR1, xenotropic and polytropic retrovirus receptor) domain proteins have been shown to be involved in the phosphate-related signal transduction and regulation pathways. Recently, Arabidopsis AtSPX family genes have been found to possess diverse functions in plant tolerance to phosphorus starvation, and OsSPX1 is involved in phosphate homeostasis in rice and optimizes growth under phosphate-limited conditions through a negative feedback loop. In this study, our phylogenetic and gene expression profiling approaches identified six rice OsSPX genes up-regulated during cold stress. Transgenic tobacco plants with constitutive expression of OsSPX1 were more tolerant to cold stress than were wild-type plants, and showed better seedling survival and reduced cellular electrolyte leakage. In addition, there was decreased total leaf Pi content and accumulation of free proline and sucrose in transgenic tobacco plants during cold stress. To further establish a cause-and-effect relationship between intracellular Pi level and cold acclimatization in transgenic plants, we generated transgenic Arabidopsis plants with constitutive expression of OsSPX1. Cold stress resulted in reduced leaf Pi levels in Arabidopsis transgenic relative to wild-type plants. From real-time reverse transcriptase-polymerase chain reaction analysis, several Pi starvation-related genes, such as AtSPX1 (orthologue of OsSPX1), PHO2, PLDZ2 and ATSIZ1, showed differential expression between wild-type and transgenic plants during cold stress. Our results indicate that OsSPX1 may play an important role in linking cold stress and Pi starvation signal transduction pathways.


The critic's voice: On the role and function of criticism of classical music recordings.
In the Western classical tradition music criticism represents one of the most complex and influential forms of performance assessment and evaluation. However, in the age of peer opinion sharing and quick communication channels it is not clear what place music critics' judgments still hold in the classical music market. This article presents expert music critics' view on their role, function, and influence. It is based on semi-structured interviews with 14 native English- and German-speaking critics who had an average of 32 years professional activity in classical music review. We present the first visual model to summarize music critics' descriptions of their role and responsibilities, writing processes, and their influences (on the market and on artists). The model distinguishes six roles (hats): consumer adviser, teacher, judge, writer, stakeholder, and artist advocate. It identifies core principles governing critical writing for music as well as challenges that arise from balancing the above six responsibilities whilst remaining true to an implicit code of conduct. Finally, it highlights the factors that inform critics' writing in terms of the topics they discuss and the discursive tools they employ. We show that music critics self-identify as highly skilled mediators between artists, producers and consumers, and justify their roles as judge and teacher based on a wealth of experience as against the influx of pervasive amateur reviews. Our research approach also offers occupation-based insights into professional music review standards, including the challenges of maintaining objectivity and resisting commercial pressures. This article offers a new viewpoint on music critics' judgments and recommendations that helps to explain their expectations and reflections.