Seeking sunlight: rapid phototactic motility of filamentous mat-forming cyanobacteria optimize photosynthesis and enhance carbon burial in Lake Huron's submerged sinkholes.
We studied the motility of filamentous mat-forming cyanobacteria consisting primarily of Oscillatoria-like cells growing under low-light, low-oxygen, and high-sulfur conditions in Lake Huron's submerged sinkholes using in situ observations, in vitro measurements and time-lapse microscopy. Gliding movement of the cyanobacterial trichomes (100-10,000 μm long filaments, composed of cells ∼10 μm wide and ∼3 μm tall) revealed individual as well as group-coordinated motility. When placed in a petri dish and dispersed in ground water from the sinkhole, filaments re-aggregated into defined colonies within minutes, then dispersed again. Speed of individual filaments increased with temperature from ∼50 μm min(-1) or ∼15 body lengths min(-1) at 10°C to ∼215 μm min(-1) or ∼70 body lengths min(-1) at 35°C - rates that are rapid relative to non-flagellated/ciliated microbes. Filaments exhibited precise and coordinated positive phototaxis toward pinpoints of light and congregated under the light of foil cutouts. Such light-responsive clusters showed an increase in photosynthetic yield - suggesting phototactic motility aids in light acquisition as well as photosynthesis. Once light source was removed, filaments slowly spread out evenly and re-aggregated, demonstrating coordinated movement through inter-filament communication regardless of light. Pebbles and pieces of broken shells placed upon intact mat were quickly covered by vertically motile filaments within hours and became fully buried in the anoxic sediments over 3-4 diurnal cycles - likely facilitating the preservation of falling debris. Coordinated horizontal and vertical filament motility optimize mat cohesion and dynamics, photosynthetic efficiency and sedimentary carbon burial in modern-day sinkhole habitats that resemble the shallow seas in Earth's early history. Analogous cyanobacterial motility may have played a key role in the oxygenation of the planet by optimizing photosynthesis while favoring carbon burial.


Sonographic value in diagnosis of hemorrhagic ovarian cysts.
To investigate the sonographic characteristics of hemorrhagic ovarian cysts (HOC) and to avoid unnecessary surgery. 113 cases of suspected HOC underwent sonographic and clinical follow-up for three months. 104 cases were clinically diagnosed with HOC as the masses disappeared naturally. The mean length of the greatest diameter was 5.12 +/- 1.33 cm, and the mean period of disappearance was 3.5 +/- 2.4 weeks. There were four patterns of the image: 21 cases (20.2%) showed a diffused dense echo pattern, 25 cases (24.0%) displayed a mixed pattern, 30 cases (28.8%) expressed a sponge-like pattern and 28 cases (27.0%) exhibited a cystic pattern. Ring blood flow with high velocity and low resistance was detected in 41 cases (40%) and there was no internal blood flow. HOC showed characteristic features on sonography, which provided useful information to differentiate HOC from ovarian tumors.


Curcumin induces apoptosis and autophagy inhuman renal cell carcinoma cells via Akt/mTOR suppression.
Renal cell carcinoma (RCC) is a highly aggressive cancer leading to high economic and social burden, and has increasing annual cases. Curcumin is a traditional Chinese medicine widely used as anti-inflammatory, anti-viral and anti-cancer agent, thus can be applicable in RCC therapy. The work assessed the effects of RCC treatment with Curcumin, Curcumin+3-MA, Curcumin+ CQ or curcumin+ Z-VAD in vitro and in vivo, and the mechanisms involved in inhibition of tumor cells proliferation. The study used ACHN tumor cells and C57BL/6 nude mice for results validation. Cell proliferation was determined through MTT assays while apoptosis was investigated using Annexin V-FITC/PI kit and flow cytometry. Enzyme-linked immunosorbent assay (ELISA) was used to detect IL-6, IL-8, and TNF-α cytokines expressions. AKT/mTOR and autophagy proteins expressions were investigated through western blot and immunofluorescence. The results indicated significantly inhibited cell viability following ACHN tumor cells treatments with curcumin alone, or with the various combinations, as compared to the control. Apoptosis was significantly increased following curcumin treatment, but was significantly reversed after treatment with curcumin+ 3-MA. Likewise, AKT/mTOR proteins expression were significantly reduced while the autophagy-related proteins were significantly elevated following curcumin treatment. The tumor size, weight and volumes were also significantly suppressed following treatment with curcumin. In conclusion, the investigation demonstrated that curcumin suppressed ACHN cell viability, induced apoptosis and autophagy, through the suppression of AKT/mTOR pathway. Use of curcumin to target AKT/mTOR pathway could be an effective treatment alternative for renal cell carcinoma.


Evaluating the fit of ambidextrous and fitted gloves: implications for hand discomfort.
Ambidextrous gloves stretch significantly more and exert greater force than fitted gloves when worn. Over time, this force could contribute to vascular constriction, nerve compression, muscle fatigue and hand pain. This study suggests that research on the effects of glove use by dental care personnel is needed.


[Perception of barriers and facilitators for users to participate in physical activity programs].
In Brazil, physical activity is a priority area in health promotion in the Brazilian Unified National Health System (SUS). The study aims to describe users' profiles and the barriers and facilitators in basic healthcare for participation in programs to promote physical activity. This was a statewide cross-sectional study in municipalities (counties) in Pernambuco State. Between 2 and 21 individuals were interviewed per municipality, using a previous tested instrument, validated in the following dimensions: sociodemographic; health status; participation; information on the program; reasons for participation; facilitators; barriers; and preference of activities. Descriptive and inferential analyses (chi-square) were performed. The sample consisted of 1,153 users, 35.9% of whom between 41-59 years of age; 90.1% lived in urban areas; 58.2% had participated in physical activities for at least a year. Weekly frequency of 3-4 days was 44.9%, and 71.1% practiced activities of an hour or more. The results showed that 40% of the barriers to participation in physical activity programs and 77.5% of the facilitators belonged to the intrapersonal domain. The most prevalent barrier was "current health condition" and the facilitator was the desire "to be healthier". Women perceived more barriers than men. The conclusion is that barriers and facilitators from the intrapersonal domain, related to health, are the main factors involved in users' engagement and retention in programs and interventions to promote physical activity, developed in basic healthcare services in the state of Pernambuco.


The mutagenic potencies of plant extracts containing quercetin in Salmonella typhimurium TA98 and TA100.
Four commercial ethanolic plant extracts, Tinctura Alchemillae, Extractum Crataegi, Extractum Myrtilli and Tinctura Hyperici, were tested for their mutagenicity in Salmonella typhimurium TA98 and TA100 with and without S9 mix obtained from rats pretreated with phenobarbital. The extracts studied differed greatly in their mutagenic potencies but exhibited a very similar mutation pattern in which the strongest effect was always seen in tester strain TA98 with S9 mix. Simultaneously we investigated the extracts for the presence of quercetin and kaempferol. Only quercetin was detected in small amounts by thin-layer chromatography (TLC). The fractions containing quercetin were separated and collected using a Sephadex LH-20 column. Two different methods were employed to estimate the amount of quercetin in the extracts: a colorimetric assay developed by Christ and Müller, and a complexometric method by Belikov. The quercetin concentrations ranged between 2 mg (Tinctura Alchemilla) and 89 mg (Tinctura Hyperici) per 100 g of extract. We suggest that the mutagenicity of the 4 plant extracts is mainly due to the presence of quercetin for the following reasons: (1) all the plant extracts exhibit a mutation pattern which is very similar to that of quercetin, (2) the mutagenic potential of the extracts correlates well with their quercetin content, considering the fact that plant extracts are very complex mixtures often containing toxic or antimutagenic compounds.


A modified technique of ureteroplasty for megaureter in children.
Excisional ureteroplasty carries the risks of jeopardizing the ureteral vasculature and leakage from the suture. The folding techniques are theoretically less prone to these risks, although they have other disadvantages due to the bulky ureter. According to the literature, these 2 approaches have similar complication rates of 4% to 25%, to include stenosis, reflux and leakage. We introduce a modified ureteroplasty technique with the aim of ensuring effective reduction of the ureteral diameter with minor risks to the vasculature. A total of 42 consecutive patients underwent ureteroplasty and reimplantation (Cohen 16, Politano-Leadbetter 3, psoas hitch 23) between 1994 and 2004, and were followed for 1 to 9 years. The ureter was opened longitudinally on its less vascularized area. Two parallel longitudinal incisions were made from the luminal side up to the musculature layer, leaving the adventitia untouched. The mucosal aspects lateral to these lines were discarded. The inner layer was closed with a running suture. The adventitial layer was closed with single stitches. No leakage, stenosis or reflux was observed. In 3 ureters persistent dilatation was observed, without obstruction or reflux. Our modification combines some principles of the 2 classic techniques, with the purpose of decreasing the risks and disadvantages of both. We believe that our approach affords better preservation of the ureteral vasculature because the adventitia is preserved untouched, as well as effective caliber reduction so that the bulking problem is avoided. In addition, the technique is associated with a minor risk of leakage. Our results show that this approach is a valid option for megaureter correction in children.


Clinicopathological and prognostic significance of FOXP3+ tumor infiltrating lymphocytes in patients with breast cancer: a meta-analysis.
The prognostic significance of FOXP3+ tumor-infiltrating lymphocytes (TILs) in patients with breast cancer remains controversial. The aims of our meta-analysis are to evaluate its association with clinicopathological characteristics and prognostic significance in patients with breast cancer. PubMed, Embase, Cochrane Database and the Ovid Database were systematically searched (up to April 2015). The meta-analysis was performed using hazard ratio (HR), odds ratio (OR) and 95 % confidence intervals (CI) as effect measures. Using the random-effects model, statistical analysis was performed using Stata software, version 12.0. Seventeen studies including 8277 patients with breast cancer were analyzed. The meta-analysis indicated that the incidence difference of FOXP3+ TILs was significant when comparing the lymph node positive group to negative group (OR = 1.305, 95 % CI [1.071, 1.590]), the histological grade III group to the I-II group (OR = 3.067, 95 % CI [2.288, 4.111]), the ER positive group to the negative group (OR = 0.435, 95 % CI [0.287, 0.660]), the PR positive group to the negative group (OR = 0.493, 95 % CI [0.296, 0.822]), the HER2 positive group to the negative group (OR = 1.896, 95 % CI [1.335, 2.692]), the TNBC group to the non TNBC group (OR = 2.456, 95 % CI [1.801, 3.348]). The detection of FOXP3+ TILs was significantly correlated with the recurrence-free survival (RFS) of patients (HR = 1.752, 95 % CI [1.188-2.584]) and the overall survival (OS) of patients (HR =1.447, 95 % CI [1.037-2.019]). Our meta-analysis demonstrates that the presence of high levels of FOXP3+ TILs is associated with prognosis for breast cancer patients and predicts lymph node metastasis, hormone receptor and HER-2 status.


[The biochemistry of alcohol metabolism].
The metabolism of ethanol to acetaldehyde in the liver proceeds via alcohol dehydrogenase (ADH) and the microsomal ethanol-oxidizing system (MOS), whereas catalase plays no significant role. ADH is an enzyme of the cytosol, requires NAD+ as cofactor and exhibits a pH optimum in the alkaline range. The Km of ADH is about 2 mM for ethanol (equivalent to 0.1%). Thus, the enzyme is already saturated at low ethanol concentrations. Conversely, MEOS resides in the endoplasmic reticulum, requires NADPH and O2, is inhibited by CO and exhibits a km of about 10 mM corresponding to 0.5% ethanol. This enzyme system is therefore primarily the pathway of ethanol metabolism at intermediate to high ethanol concentrations. MEOS has many properties in common with other drug metabolizing enzymes and is characterized by inducibility following chronic ethanol consumption, which suggests the involvement of the microsomal system in the adaptive enhancement of ethanol clearance commonly observed in alcoholics. The product of ethanol oxidation by ADH, MEOS and catalase is acetaldehyde. Acetaldehyde is oxidized in the liver to acetate by NAD dependent aldehyde dehydrogenase. Four isozymes have been identified. Lack of isozyme I is responsible for the "flush-syndrome" commonly observed in asian people following alcohol intake. Ethanol metabolism is affected by the aging process and is decreased with advancing age.


Carbapenem-resistant Klebsiella pneumoniae in high-risk haematological patients: factors favouring spread, risk factors and outcome of carbapenem-resistant Klebsiella pneumoniae bacteremias.
Carbapenem-resistant Klebsiella pneumoniae (CRKP) spread and infections in patients with haematological malignancies are a serious concern especially in endemic areas. Treatment failures and delay in appropriate therapy for CRKP infections are frequent and the mortality rate associated with CRKP bacteremia in neutropenic haematological patients is reported about 60%. Haematological patients harboring CRKP hospitalized between February 2012 and May 2013 in an Italian Teaching hospital were examined. Conditions favouring CRKP spread in a haematological unit, risk factors for bacteremia in CRKP-carriers and for CRKP bacteremia-related death were evaluated in this observational retrospective study. CRKP was isolated in 22 patients, 14 (64%) had bacteremia. Control measures implementation, particularly the weekly rectal screening for CRKP performed in all hospitalized patients and contact precautions for CRKP-carriers and newly admitted patients until proved CRKP-negative, reduced significantly the CRKP spread (14 new carriers identified of 131 screened patients vs 5 of 242 after the intervention, p = 0.001). Fifty-eight percent of carriers developed CRKP bacteremia, and acute myeloid leukemia (AML) resulted independently associated with the bacteremia occurrence (p = 0.02). CRKP bacteremias developed mainly during neutropenia (86%) and in CRKP-carriers (79%). CRKP bacteremias were breakthrough in 10 cases (71%). Ten of 14 patient with CRKP bacteremias died (71%) and all had AML. The 70% of fatal bacteremias occurred in patients not yet recognized as CRKP-carriers and 80% were breakthrough. Initial adequate antibiotic therapy resulted the only independent factor able to protect against death (p = 0.02). The identification of CRKP-carriers is confirmed critical to prevent CRKP spread. AML patients colonized by CRKP resulted at high risk of CRKP-bacteremia and poor outcome and the adequacy of the initial antibiotic therapy may be effective to improve survival. To limit the increase of resistance, the extensive use of antibiotics active against CRKP should be avoided, but in the setting of high CRKP pressure and high-risk CRKP-colonized haematological patients, timely empiric antibiotic combinations active against CRKP could be suggested as treatment of febrile neutropenia.


Procedural learning of semantic categorization in Parkinson's disease.
This paper studies the procedural learning of semantic categorization in 29 patients with non-demented Parkinson's disease (PD). We investigated whether the PD group was able to develop semantic skill, using a cognitive procedural task developed in our laboratory, applying a manual and serial reaction time paradigm to semantic categorization. The PD group showed similar scores to those of the control group on semantic categorization. Both groups showed reaction time reduction over the semantic procedural task, but the PD group produced longer reaction times than the control subjects. Contrary to our prediction, we observed an improvement in semantic categorization reaction times with practice, even with new verbal material for the PD patients to categorize despite their motor impairments and executive deficits. By contrast, we found a significant negative correlation between axial motor signs and the ratio of semantic procedural learning, but not for lateral motor signs. The present results support the notion that non-demented PD patients may be capable of acquiring comparable semantic skill to those of the control group.


The role of cervical pedicle screw in cervical spine trauma: A single-center retrospective study.
Placement of pedicle screw in the subaxial cervical spine is a challenging and complex technique but provides significant biomechanical advantages. Despite its potential complications, the role and use of cervical pedicle screw (CPS) are growing. A literature review of the significant articles on applying pedicle screws in the subaxial cervical spine was done (articles between 1994 and 2020). Furthermore, our center´s experience of 15 years related to CPS is also discussed in this study. Transpedicular instrumentation in the subaxial cervical spine requires profound anatomical knowledge and meticulous surgical technique. This technique provides superior biomechanical stability compared to the other cervical fixation techniques. Pull-out strength of CPS is twice as compared to the lateral mass screws. There have been numerous variations in the technique of CPS, varying from open techniques to minimally invasive and the use of biomodels and templates during this procedure. Clinically, CPS can be used in different cervical trauma situations, such as fracture-dislocations, floating lateral mass, and fractures associated with ankylosing spondylitis. Despite the possibility of neurovascular injury due to the proximity of the vertebral artery, spinal cord, and spinal nerves to the cervical pedicles, scientific literature, and our center × s experience show low risk, and this technique can be performed safely. CPS placement is a safe procedure, and it has great potential in the management of cervical spine trauma.


Variations in survival after cardiac arrest among academic medical center-affiliated hospitals.
Variation exists in cardiac arrest (CA) survival among institutions. We sought to determine institutional-level characteristics of academic medical centers (AMCs) associated with CA survival. We examined discharge data from AMCs participating with Vizient clinical database-resource manager. We identified cases using ICD-9 diagnosis code 427.5 (CA) or procedure code 99.60 (CPR). We estimated hospital-specific risk-standardized survival rates (RSSRs) using mixed effects logistic regression, adjusting for individual mortality risk. Institutional and community characteristics of AMCs with higher than average survival were compared with those with lower survival. We analyzed data on 3,686,296 discharges in 2012, of which 33,700 (0.91%) included a CA diagnosis. Overall survival was 42.3% (95% CI 41.8-42.9) with median institutional RSSR of 42.6% (IQR 35.7-51.0; Min-Max 19.4-101.6). We identified 28 AMCs with above average survival (median RSSR 61.8%) and 20 AMCs with below average survival (median RSSR 26.8%). Compared to AMCs with below average survival, those with high CA survival had higher CA volume (median 262 vs.119 discharges, p = 0.002), total beds (722 vs. 452, p = 0.02), and annual surgical volume (24,939 vs. 13,109, p<0.001), more likely to offer cardiac catheterization (100% vs. 72%, p = 0.007) or cardiac surgery (93% vs. 61%, p = 0.02) and cared for catchment areas with higher household income ($61,922 vs. $49,104, p = 0.004) and lower poverty rates (14.6% vs. 17.3%, p = 0.03). Using discharge data from Vizient, we showed AMCs with higher CA and surgical case volume, cardiac catheterization and cardiac surgery facilities, and catchment areas with higher socioeconomic status had higher risk-standardized CA survival.


Can nutritional information modify purchase of ultra-processed products? Results from a simulated online shopping experiment.
The aim of the present work was to evaluate the influence of two front-of-pack nutrition information schemes (traffic-light system and Chilean warning system) on consumer purchase of ultra-processed foods in a simulated online grocery store. Following a between-subjects design, participants completed a simulated weekly food purchase in an online grocery store under one of three experimental conditions: (i) a control condition with no nutrition information, (ii) a traffic-light system and (iii) the Chilean warning system. Information about energy (calories), sugar, saturated fats and salt content was included in the nutrition information schemes. Participants were recruited from a consumer database and a Facebook advertisement. People from Montevideo (Uruguay), aged 18-77 years (n 437; 75 % female), participated in the study. All participants were in charge of food purchase in the household, at least occasionally. No significant differences between experimental conditions were found in the mean share of ultra-processed foods purchased by participants, both in terms of number of products and expenditure, or in the mean energy, sugar, saturated fat and salt content of the purchased items. However, the Chilean warning system decreased intended purchase of sweets and desserts. Results from this online simulation provided little evidence to suggest that the traffic-light system or the Chilean warning system in isolation could be effective in reducing purchase of ultra-processed foods or improving the nutritional composition of the purchased products.


New Applications for Submucosal Tunneling in Third Space Endoscopy: A Comprehensive Review.
Third space endoscopy (TSE), a type of natural orifice transluminal endoscopic surgery, was first introduced a decade ago, and since then, has revolutionized the field of therapeutic endoscopy. TSE involves tunneling into the submucosal space between the intestinal mucosa and the muscularis propria. By accessing this "third space," endoscopists can perform minimally invasive myotomies, resections, and dissections while maintaining the mucosa and preventing full-thickness perforation. TSE procedures can be used to treat motility disorders as well as neoplasms throughout the gastrointestinal tract, which were traditionally conditions that would require open or laparoscopic surgery. The main TSE procedures include peroral endoscopic myotomy, gastric peroral endoscopic myotomy, Zenker peroral endoscopic myotomy, submucosal tunneling for endoscopic resection, endoscopic submucosal tunnel dissection, peroral endoscopic tunneling for restoration of the esophagus, and per-rectal endoscopic myotomy. Some of those procedures have now been accepted as viable alternative to surgical techniques due to their limited invasiveness, safety and cost. This review will explore the different techniques recently added to our arsenal while describing their potential efficacy and limitations.


Deciphering the fine details of c1 assembly and activation mechanisms: "mission impossible"?
The classical complement pathway is initiated by the large (~800 kDa) and flexible multimeric C1 complex. Its catalytic function is triggered by the proteases hetero-tetramer C1r2s2, which is associated to the C1q sensing unit, a complex assembly of 18 chains built as a hexamer of heterotrimers. Initial pioneering studies gained insights into the main architectural principles of the C1 complex. A dissection strategy then provided the high-resolution structures of its main functional and/or structural building blocks, as well as structural details on some key protein-protein interactions. These past and current discoveries will be briefly summed up in order to address the question of what is still ill-defined. On a functional point of view, the main molecular determinants of C1 activation and its tight control will be delineated. The current perspective remains to decipher how C1 really works and is controlled in vivo, both in normal and pathological settings.


PSD-95 deficiency disrupts PFC-associated function and behavior during neurodevelopment.
Postsynaptic density protein-95 (PSD-95) is a major regulator in the maturation of excitatory synapses by interacting and trafficking N-methyl-D-aspartic acid receptors (NMDAR) and α-amino-3-hydroxy-5-methyl-4-isox-azoleproprionic acid receptors (AMPAR) to the postsynaptic membrane. PSD-95 disruption has recently been associated with neuropsychiatric disorders such as schizophrenia and autism. However, the effects of PSD-95 deficiency on the prefrontal cortex (PFC)-associated functions, including cognition, working memory, and sociability, has yet to be investigated. Using a PSD-95 knockout mouse model (PSD-95-/-), we examined how PSD-95 deficiency affects NMDAR and AMPAR expression and function in the medial prefrontal cortex (mPFC) during juvenile and adolescent periods of development. We found significant increases in total protein levels of NMDAR subunits GluN1, and GluN2B, accompanied by decreases in AMPAR subunit GluA1 during adolescence. Correspondingly, there is a significant increase in NMDAR/AMPAR-mediated current amplitude ratio that progresses from juvenile-to-adolescence. Behaviorally, PSD-95-/- mice exhibit a lack of sociability, as well as learning and working memory deficits. Together, our data indicate that PSD-95 deficiency disrupts mPFC synaptic function and related behavior at a critical age of development. This study highlights the importance of PSD-95 during neurodevelopment in the mPFC and its potential link in the pathogenesis associated with schizophrenia and/or autism.


[Investigations concerning bovine gastro-intestinal strongyles. II. Seasonal variations in the herbage infestation with infective larvae (author's transl)].
Ecological studies of bovine gastro-intestinal trichostrongylide larvae have been carried on during a three-year period (1972-1974). The studies mainly included pastures grazed by heifers (2nd time grazing cattle). Examination of the herbage samples was based on washing and filtration (sieve mesh-size 36 and 28 mu) followed by Baermann test on the debris. The pasture infestation with infective larvae (L3) followed a nearly stereotype seasonal pattern (cf. fig. 2 and fig. 3): A considerable larval population survived on the pasture during the winter, the infestation decreased strongly during April-May, was low during June and beginning of July, and increased from the middle of July to reach a peak in September-October. The infestation was usually much higher just around the faecal pats than in the rest of the pasture area (cf. fig. 3). The observed populations of trichostrongylide larvae were dominated by Ostertagia (ostertagi), but included other species as well, e.g. Cooperia (oncophora), Nematodirus spp., etc.


Altered folate receptor 2 expression in uraemic patients on haemodialysis: implications for folate resistance.
Folate therapy reduces, but does not normalize homocysteine (Hcy) levels, frequently elevated in chronic kidney disease (CKD). The mechanisms of this folate resistance are unknown. Cellular acquisition of folate is mediated by folate receptors (FRs), whose expression is also modulated by folate status, through an Hcy-dependent regulation mechanism involving heterogeneous nuclear ribonucleoprotein-E1 (hnRNP-E1). Our objective was to evaluate whether an alteration of the FR2 (the form present in nucleated blood cells) expression is present in CKD patients on haemodialysis (HD), and its susceptibility to folate treatment. A population of chronic uraemic patients on HD was enrolled, along with a control group, and studies on FR2 receptor expression and related items were performed in plasma and mononuclear cells from peripheral blood. A subgroup of patients was treated with methyltetrahydrofolate for 1 month. In HD, there was a significant reduction in FR2 protein expression compared with controls, not correlated with Hcy concentrations, while its mRNA levels were significantly increased. After folate treatment, there was a significant mRNA decrease, in the absence of significant changes in receptor protein expression. hnRNP-E1 gene and protein expression levels increased pre-treatment, while decreased post-treatment. In HD, FR2 expression is altered in peripheral mononuclear cells, since its levels are decreased and are not responsive to variations in Hcy concentration, while the intracellular machinery (receptor mRNA and hnRNP-E1), possibly triggering its regulation, is conserved. These findings provide insight into the mechanisms of folate resistance in uraemia.


Heart disease in a mutant mouse model of spontaneous eosinophilic myocarditis maps to three loci.
Heart disease (HD) is the major cause of morbidity and mortality in patients with hypereosinophilic diseases. Due to a lack of adequate animal models, our understanding of the pathophysiology of eosinophil-mediated diseases with heart complications is limited. We have discovered a mouse mutant, now maintained on an A/J inbred background, that spontaneously develops hypereosinophilia in multiple organs. Cellular infiltration into the heart causes an eosinophilic myocarditis, with affected mice of the mutant line (i.e., A/JHD) demonstrating extensive myocardial damage and remodeling that leads to HD and premature death, usually by 15-weeks old. Maintaining the A/JHD line for many generations established that the HD trait was heritable and implied the mode of inheritance was not too complex. Backcross and intercross populations generated from mating A/JHD males with females from four different inbred strains produced recombinant populations with highly variable rates of affected offspring, ranging from none in C57BL/6 J intercrosses, to a few mice with HD using 129S1/SvImJ intercrosses and C57BL/6 J backcrosses, but nearly 8% of intercrosses and > 17% of backcrosses from SJL/J related populations developed HD. Linkage analyses of these SJL/J derived recombinants identified three highly significant loci: a recessive locus mapping to distal chromosome 5 (LOD = 4.88; named Emhd1 for eosinophilic myocarditis to heart disease-1); and two dominant variants mapping to chromosome 17, one (Emhd2; LOD = 7.51) proximal to the major histocompatibility complex, and a second (Emhd3; LOD = 6.89) that includes the major histocompatibility region. Haplotype analysis identified the specific crossovers that defined the Emhd1 (2.65 Mb), Emhd2 (8.46 Mb) and Emhd3 (14.59 Mb) intervals. These results indicate the HD trait in this mutant mouse model of eosinophilic myocarditis is oligogenic with variable penetrance, due to multiple segregating variants and possibly additional genetic or nongenetic factors. The A/JHD mouse model represents a unique and valuable resource to understand the interplay of causal factors that underlie the pathology of this newly discovered eosinophil-associated disease with cardiac complications.


Twenty-one years of microemulsion electrokinetic chromatography (1991-2012): a powerful analytical tool.
Microemulsion electrokinetic chromatography (MEEKC) is a CE separation technique, which utilizes buffered microemulsions as the separation media. In the past two decades, MEEKC has blossomed into a powerful separation technique for the analysis of a wide range of compounds. Pseudostationary phase composition is so critical to successful resolution in EKC, and several variables could be optimized including surfactant/co-surfactant/oil type and concentration, buffer content, and pH value. Additionally, MEEKC coupled with online sample preconcentration approaches could significantly improve the detection sensitivity. This review comprehensively describes the development of MEEKC from the period 1991 to 2012. Areas covered include basic theory, microemulsion composition, improving resolution and enhancing sensitivity methods, detection techniques, and applications of MEEKC.


Evaluation of Guamanian dental caries preventive programs after 13 years.
To address the high prevalence of dental caries in Guamanian children, a school-based fluoride mouthrinse program, a clinic-based pit and fissure sealant program, and community water fluoridation were phased in over a 13-year period. The purpose of this paper is to evaluate the impact of these programs on the prevalence of dental caries. Cross-sectional samples representative of schoolchildren in grades 1 through 8 were selected in 1976, 1979, 1984, 1986, and 1989. These years include the starting years for each of the three interventions and varying follow-up periods for each. About 1,000 children in each of the survey years were examined to derive DMF surface scores. After eight years of fluoride mouthrinsing, DMFS scores were reduced by 25.4 percent in 6-14-year-olds. With two additional years of fluoride mouthrinsing and with pit and fissure sealants, overall DMFS scores declined an additional 44.4 percent. In 1989, three years after community water fluoridation was initiated on the island and continuation of the other two programs, there was a further decline in overall DMFS scores of 34.5 percent. Over the entire study period--during which there were 13 years of fluoride mouthrinsing in the schools, five years of sealant application, and three years of community water fluoridation--DMFS scores declined 72.8 percent overall (5.14 surfaces per child) and 71.9, 71.0, and 78.8 percent for occlusal, buccal-lingual, and proximal surfaces, respectively. An intensified preventive dentistry program introduced on an island with high caries prevalence twice that of the US mainland was successful, and contributed to a reduction in the prevalence of caries to a level equivalent to that of the United States at the end of the study period.


New insights into gene positional clustering and its properties supported by large-scale analysis of various differentiation pathways.
To understand how genes are distributed on chromosomes we bring new insights into gene positional clustering and its properties. We have made a large-scale analysis of three types of differentiation and we observed that genes that subsequently enter into different cell processes are positionally clustered on chromosomes. Genes from the clusters are transcribed subsequently with respect to time kinetics and also to position. This means that the genes related to a cellular process are clustered together, independent of the period of time during which they are active and important for the process. Our results also demonstrate not only that there are general regions of increased or decreased levels of gene expression, but also that, in fact, in some chromosome regions we can find clustering of genes related to specific cell processes. The results provided in this paper also support the theory of "transcription factories" and show that transcription of genes from the clusters is managed by softer epigenetic mechanisms.


Potent suppression of arginase 1 expression in murine macrophages by low dose endotoxin.
Macrophages can respond to diverse signals and adopt multiple phenotypes. Although interleukin-4 (IL-4) is shown to potently induce the expression of arginase 1 and contribute to differentiation of macrophages to the anti-inflammatory M2 phenotype, other modulators may potentiate or reduce the effect of IL-4. In this report, we focus on the combinatorial effects of IL-4 with all-trans retinoic acid (ATRA) and lipopolysaccharide (LPS). ATRA has been shown to contribute to the resolution of inflammation, however it has not been linked to arginase 1 expression in macrophages. We demonstrate that although ATRA alone has no effect on the expression or activities of arginase 1, ATRA can dramatically potentiate the induction of arginase 1 by IL-4. On the other hand, high doses of LPS, such as those seen in septic shock, can induce the expression of both M1 and M2 mediators in macrophages. The effects of subclinical doses of LPS, which are prevalent in humans with adverse health conditions, on macrophage differentiation are not well studied. We demonstrate that low dose LPS can effectively suppress the expression of arginase 1 induced by IL-4 and ATRA. Mechanistically, we report that the interleukin-1 receptor-associated kinase 1 (IRAK-1) and Toll-interacting-protein (Tollip) are involved in the suppressive effect of low dose LPS. Our study reveals dynamic modulation of arginase 1 expression in macrophages by competing agonists, and bears relevance for potential intervention of chronic diseases.


Physiological responses to exercise in patients following fracture of the lower limb.
Twenty-five patients with healed fractures of the lower limb and nine normal control subjects were measured anthropometrically and during maximal and submaximal one- and two-leg bicycle exercise. Oxygen intake at a given submaximal work level of 450 kmp min-1 and cardiac frequency at an oxygen intake of 1.5 I min-1 were significantly higher (p less than 0.001) in the injured compared with the uninjured limbs of the patients and normal subjects. The maximum aerobic power of the injured and uninjured limbs of the patients and normal subjects. The maximum aerobic power of the injured and uninjured limbs of the patients were 18.8% (0.44 I min-1) and 25.6% (0.61 I min-1) respectively lower than the right and left legs of the control subjects. The corresponding value for 2-leg work was 17.6% (0.51 I min-1). The deterioration in 1-leg performance of the patients was associated with a concomitant decrease in leg muscle (plus bone) volume. In 2-leg work this factor was also probably combined with a deterioration in performance due to general cardiovascular deconditioning.


One day is better than four days of ejaculatory abstinence for sperm function.
According to the World Health Organization guidelines, ejaculatory abstinence (EA) of 2-7 days is recommended for semen analysis. This study aimed to determine how seminal quality may be affected by two EA periods from the same man. Seminal samples from 65 men were evaluated by conventional semen analysis and qualitative characteristics after 1 and 4 days of EA (two samples/man). The semen was qualitatively analyzed by examining oxidative activity (intracellular and seminal plasma), sperm function (acrosome integrity, mitochondrial activity, and nuclear DNA integrity), and epididymal function. As expected, samples collected after 1 day of EA showed a decrease in volume and sperm total number compared to samples collected after 4 days of EA. The sperm motility of the samples collected after 1 day of EA was better compared to samples collected after 4 days of EA. Oxidative activity measured was lower after 1 day of EA compared with those measured after 4 days of EA. With regards to sperm function, samples collected after 1 day of EA showed an increase in acrosome integrity, mitochondrial activity, and nuclear DNA integrity compared with samples collected after 4 days of EA. Epididymal function showed no difference between the two-time points. Although samples collected after 4 days of EA showed better results for sperm quantity, samples collected after 1 day of EA showed better qualitative results, including motility, oxidative activity, and sperm function. Thus, it can be concluded that sperm storage at the epididymal tail may make spermatozoa more susceptible to oxidative damage. According to the World Health Organization guidelines, stopping ejaculation for 2 to 7 days is recommended before sperm collection for semen analysis. However, the evidence that supports these recommendations is limited. Our study aimed to compare how sperm quality was affected in samples collected after stopping ejaculation for 1 day and 4 days (two samples per man) in a total of 65 men. Although sample collection after stopping ejaculation for 4 days showed better semen quantity (volume and sperm concentration), sample collection after stopping ejaculation for 1 day showed better sperm motility and function. If not ejaculated, sperm are stored in the epididymis tail located in the scrotum beside the testicles and our study suggests that longer sperm storage may damage sperm quality. The results from this study may be used to inform guidance for sperm collection for use in assisted reproduction techniques, and lead to an improvement in both fertilization and implantation rates.


Parents' knowledge of and opinions about healthcare laws and technology in primary care.
Historically, parents have demonstrated poor understanding of adolescent healthcare laws. This study assessed US parents' current knowledge and opinions about technology facilitated physician-adolescent communication and applicable laws to enhance transition to adult health care. A brief survey in two low-income academic paediatric clinics asked parents about their knowledge of health care and laws, and their opinions about technology facilitated contact between physicians and adolescents. Almost all parents (96.7%) have internet access at home, work or via a mobile device. Only 44.1% approved of having a physician directly contact their child about annual examinations, immunisations or to discuss issues of sexuality. Half (55.4%) were aware that adolescents could receive confidential sexuality information and treatment without parents' permission. Only one-third (32.2%) approved of a specific technology for direct communication. Parents are divided about direct physician-adolescent contact. Future plans to engage adolescents to understand their health will require parental education and involvement on the value of physician-adolescent communication.


Differential effects of acute hypoxia and endotoxin on the secretion and expression of bone marrow interleukin-1 and interleukin-6.
Hemorrhagic shock induces tissue hypoxia and has been demonstrated to alter the myelopoietic response to bacterial lipopolysaccharide (LPS). Interleukin-1 and interleukin-6 are important mediators of immunologic events after hemorrhagic shock. Bone marrow stroma release inflammatory cytokines, which may play a role in the regulation of myelopoiesis after injury. The aim of this study was to correlate cytokine gene expression with protein release and myelopoiesis by total bone marrow cells. The role of bone marrow stroma after exposure to hypoxia and lipopolysaccharide was also examined. BALB/c mice were designated as normoxia or hypoxia and total bone marrow cells were harvested. Hypoxia mice were exposed to 2 h of 5% O2/95% N2, and then returned to room air. Additional groups of mice were given LPS intraperitoneally. Bone marrow stroma, from BALB/c mice, was similarly designated. Myelopoiesis was assessed by growth of granulocyte-macrophage progenitor cells (CFU-GM). Interleukin-1 and interleukin-6 protein activity was assessed by bioassay. RNA was extracted from both total bone marrow cells and bone marrow stroma. By day 5, LPS alone resulted in a 93% increase in CFU-GM versus normoxia. Hypoxia and LPS exposure significantly decreased CFU-GM on days 1, 3, and 5. LPS alone induced an increase in interleukin-6. At 2, 6, and 24 h, hypoxia blunted interleukin-6 release in response to LPS. Hypoxia alone could not induce interleukin-6. However, hypoxia did induce interleukin-1 mRNA without the release of bioactive protein. In the remainder of groups, interleukin-1 protein levels and mRNA levels were correlated. Bone marrow stroma interleukin-1 and interleukin-6 protein activity was consistently correlated with that of total bone marrow. These data demonstrate that bone marrow cytokine production is differentially regulated by hypoxia. Hypoxia impairs interleukin-6 protein and mRNA in response to LPS, which may play a role in the suppression of myelopoiesis after shock. Also, bone marrow stroma plays an integral role in regulating myelopoiesis.


Anti-hapten IgG antibodies bound to cell surface hapten: anti-IgG antibody prevents dissociation as measured with fluid phase hapten.
We investigated the dissociation by fluid phase hapten of IgG antibodies bound to cell surface hapten in the presence and absence of anti-IgG antibodies. Dissociation was quantitated with fluid phase hapten, preventing reassociation of the anti-hapten antibodies. More than 90% of the anti-hapten IgG alone was prevented from reassociation by low concentrations of fluid phase hapten (nanogram to microgram range). In contrast, no dissociation of some IgG-anti-IgG complexes could be measured even at 24 hr incubation in the presence of very large excess of fluid phase hapten (100 mg/ml). We excluded aggregate formation between anti-hapten antibodies due to cross-linking by anti-antibodies as a cause for decreased dissociability by 1) performing the experiments in large excess of anti-antibody, 2) showing that the phenomenon was independent of anti-hapten antibody density, 3) showing that decreased dissociation also occurred at 4 degrees C, and 4) showing that aggregation by protein A did induce decreased dissociability, albeit three orders of magnitude lower than the anti-antibody. It was concluded that anti-antibody directly affected the "avidity" of cell hapten bound anti-hapten IgG in an unknown manner.


Differential recruitment of steroid hormone response elements may dictate the expression of the pituitary gonadotropin II beta subunit gene during salmon maturation.
The role of testosterone (T) and 17 beta-estradiol (E2) in the control of chinook salmon gonadotropin II beta subunit (sGTHII beta) gene was examined. Both E2 and T specifically stimulated GTHII beta gene expression in cultured juvenile rainbow trout pituitary cells. 5'-Flanking regions of the sGTHII beta gene linked to the chloramphenicol acetyltransferase (CAT) expression vector were transfected into these pituitary cells, and cultures were treated with steroid hormones. Estrogen-stimulated CAT activity occurred with constructs containing a 13-base pair estrogen responsive element (ERE) sequence [proximal ERE (pERE)] located at -273 to -260 upstream of its transcriptional start site. Binding specificity of pERE was confirmed by mobility shift and DNA methylation interference assays using the DNA binding domain of the human estrogen receptor. Interestingly, the pERE functioned to derepress the activity of the proximal silencer (pSil) only in the pituitary cells of juvenile trout but not in cells derived from maturing and sexually matured fish. Another potential ERE sequence comprised of three tandemly linked half-ERE palindromes was located from -2736 to -2659 [distal ERE (dERE)] of the sGTHII beta gene. Distal ERE might be responsible for the steroid responsiveness of the longest sGTHII beta/CAT construct (-3500CAT) observed in the pituitary cells of maturing fish. The function of pERE and dERE were further examined in the heterologous HeLa cells by mutagenesis and cotransfection with a rainbow trout estrogen receptor expression vector. Disruption of the palindromic structure of pERE severely impaired its function. When the sequences between pERE and dERE were deleted, a 200-fold increase in CAT activity was observed in response to E2. A model is proposed to describe the regulation of GTHII beta gene expression at different reproductive stages.


Genetic association study of CTLA4 and FCεRIα polymorphisms in asthmatic patients in the southwestern region of Iran.
Asthma is a heterogeneous chronic pulmonary disease that develops due to the interaction of genetic and environmental factors. This study aimed to investigate the polymorphisms of CTLA4(SNP-318C > T, SNP + 49A > G) and FCεRIα(SNP-344T > C) genes in asthmatic patients in Southwest Iran. The study enrolled 200 patients with asthma of Arab and Bakhtiary descent and 200 healthy controls, where asthmatic patients and healthy controls were selected based on a spirometry test. Genomic DNA from whole blood samples using the TaqMan assay was used to study the genotypes of patients and healthy controls.The results indicated no statistically significant difference between cases and controls for the SNP-344C > T of the FCεR1α gene and the SNP + 49A > G, SNP-318C > T of the CTLA4 gene. There was a significant correlation between the CTLA4-318C > T allele frequency in both the case and control groups (OR = 1.83; 95%CI, 1.14-2.94; P = 0.01). We stratified genotypes according to age, gender, ethnicity, and smoking status and discovered a significant suggestive association between the SNP + 49A > G of the CTLA4 gene and smoking. Additionally, SNP + 49A > G was found to be associated with gender and age. The results indicated that the SNP-318C > T polymorphism in the CTLA4 gene might contribute to the development of asthma in the studied population. Meanwhile, smoking can exacerbate asthma in individuals with SNP + 49A > G of the CTLA4 gene.Supplemental data for this article is available online at https://doi.org/10.1080/15257770.2021.1964525 .


Microcirculatory functions in systemic sclerosis: additional parameters for therapeutic concepts?
To study the functional reactivity of the cutaneous microcirculation in progressive systemic sclerosis (PSS), hyperemic responses after arterial occlusion (3 min) and during local heating (42 degrees C) were investigated with simultaneous measurements of red blood cell flux and cutaneous oxygen tension (pcuO2) of the skin in female patients (n = 19) with PSS and in healthy female controls (n = 15). Additionally, serum levels of 6-keto-prostaglandin 1 alpha (PGF1 alpha), a stable metabolite of prostacyclin, were compared to the microcirculatory data, and both were used to evaluate further a standardized therapy with 10-d intravenous calcitonin (100 IU/d) infusion in six PSS patients. In PSS, the initial mean pcuO2 value was significantly reduced and was inversely proportional to flux and to PGF1 alpha levels, whereas the flux and pcuO2 responses to the above hyperemic stimuli showed significant reductions, revealing a pattern of "hyperemic hypoxia" probably due to exhausted functional reserves of cutaneous perfusion. During calcitonin infusion significant rises in pcuO2 and temporarily in PGF1 alpha and flux were found. After 10 d of therapy, increased pcuO2 was associated significantly with decreased flux, indicating a shifting of blood from deeper regulatory vessels to the subepidermal capillaries. Both clinical improvement and the results of microcirculatory measurements demonstrate a beneficial effect of calcitonin on the cutaneous microcirculation in PSS patients, possibly due in part to a short-term increase in release of endogenous prostacyclin from the vascular endothelium during the infusion. The disturbed reactivity of the dermal vessels in PSS is important for the evaluation of therapeutic concepts and stresses, together with the elevated PGF1 alpha plasma levels, vascular factors in the pathogenesis of PSS.


The relationship between posttraumatic growth and substance use in homeless women with histories of traumatic experience.
Posttraumatic growth (PTG)--namely, the experience of positive change in oneself or one's life following trauma--is particularly relevant for homeless women because they have greater trauma exposure than the general population, as well as higher rates of substance use. The present study examined PTG and substance use in this population. Fifty homeless women with trauma histories participated. In line with predictions, more current substance use was related to less PTG, more reliance on avoidant coping once approach coping was accounted for, and greater PTSD symptomatology. Levels of growth were comparable to those found in samples with less trauma exposure.


[Base for use a new liquid accumulation growth medium for the isolation of Salmonella from water objects].
Base for use of new luquid mild inhibitory accumulation medium for the isolation of Salmonella from water objects has been proposed. High sensitivity and efficacy of the new medium compared with routinely used magnesium and selenitic media were shown in experimental conditions. During examination of water from surface reservoirs the new accumulation medium allowed to isolate Salmonella with higher index and various serologic patterns.


Contrasting responses of photosynthesis to salt stress in the glycophyte Arabidopsis and the halophyte thellungiella: role of the plastid terminal oxidase as an alternative electron sink.
The effects of short-term salt stress on gas exchange and the regulation of photosynthetic electron transport were examined in Arabidopsis (Arabidopsis thaliana) and its salt-tolerant close relative Thellungiella (Thellungiella halophila). Plants cultivated on soil were challenged for 2 weeks with NaCl. Arabidopsis showed a much higher sensitivity to salt than Thellungiella; while Arabidopsis plants were unable to survive exposure to greater than 150 mM salt, Thellugiella could tolerate concentrations as high as 500 mM with only minimal effects on gas exchange. Exposure of Arabidopsis to sublethal salt concentrations resulted in stomatal closure and inhibition of CO2 fixation. This lead to an inhibition of electron transport though photosystem II (PSII), an increase in cyclic electron flow involving only PSI, and increased nonphotochemical quenching of chlorophyll fluorescence. In contrast, in Thellungiella, although gas exchange was marginally inhibited by high salt and PSI was unaffected, there was a large increase in electron flow involving PSII. This additional electron transport activity is oxygen dependent and sensitive to the alternative oxidase inhibitor n-propyl gallate. PSII electron transport in Thellungiella showed a reduced sensitivity to 2'-iodo-6-isopropyl-3-methyl-2',4,4'-trinitrodiphenylether, an inhibitor of the cytochrome b6f complex. At the same time, we observed a substantial up-regulation of a protein reacting with antibodies raised against the plastid terminal oxidase. No such up-regulation was seen in Arabidopsis. We conclude that in salt-stressed Thellungiella, plastid terminal oxidase acts as an alternative electron sink, accounting for up to 30% of total PSII electron flow.


Sema3A promotes the resolution of cardiac inflammation after myocardial infarction.
Optimal healing after myocardial infarction requires not only the induction of inflammation, but also its timely resolution. In patients, 30 days post myocardial infarction, circulating monocytes have increased expression of Semaphorin3A (Sema3A) as compared to directly after admission. This increased expression coincides with increased expression of Cx3CR1-a marker of non-classical monocytes that are important for immune resolution hence proper wound healing. In mice, the expression of Sema3A also increases in response to myocardial ischemia being expressed by infiltrating leukocytes. Comparing Sema3A heterozygote (HZ) and wild type (WT) mice post myocardial infarction, revealed increased presence of leukocytes in the cardiac tissues of HZ mice as compared to WT, with no differences in capillary density, collagen deposition, cardiomyocyte surface area, chemokine-or adhesion molecules expression. Whilst infarct sizes were similar 14 days after myocardial infarction in both genotypes, Sema3A HZ mice had thinner infarcts and reduced cardiac function as compared to their WT littermates. In vitro experiments were conducted to study the role of Sema3A in inflammation and resolution of inflammation as a potential explanation for the differences in leukocyte recruitment and cardiac function observed in our in vivo experiments. Here, recombinant Sema3A protein was able to affect the pro-inflammatory state of cultured bone marrow derived macrophages. First, the pro-inflammatory state was altered by the induced apoptosis of classical macrophages in the presence of Sema3A. Second, Sema3A promoted the polarization of classical macrophages to resolution-phase macrophages and enhanced their efferocytotic ability, findings that were reflected in the infarcted cardiac tissue of the Sema3A HZ mice. Finally, we demonstrated that besides promoting resolution of inflammation, Sema3A was also able to retard the migration of monocytes to the myocardium. Collectively our data demonstrate that Sema3A reduces cardiac inflammation and improves cardiac function after myocardial infarction by promoting the resolution of inflammation.


Longer-term quality of care, effectiveness, and cost-effectiveness of implementing a model of care for osteoarthritis: A cluster-randomized controlled trial.
To assess the quality of care, effectiveness, and cost-effectiveness over 12 months after implementing a structured model of care for hip and knee osteoarthritis (OA) in primary healthcare as compared to usual care. In this pragmatic cluster-randomized, controlled trial with a stepped-wedge cohort design, we recruited 40 general practitioners (GPs), 37 physiotherapists (PTs), and 393 patients with symptomatic hip or knee OA from six municipalities (clusters) in Norway. The model included the delivery of a 3-hour patient education and 8-12 weeks individually tailored exercise programs, and interactive workshops for GPs and PTs. At 12 months, the patient-reported quality of care was assessed by the OsteoArthritis Quality Indicator questionnaire (16 items, pass rate 0-100%, 100%=best). Costs were obtained from patient-reported and national register data. Cost-effectiveness at the healthcare perspective was evaluated using incremental net monetary benefit (INMB). Of 393 patients, 109 were recruited during the control periods (control group) and 284 were recruited during interventions periods (intervention group). At 12 months the intervention group reported statistically significant higher quality of care compared to the control group (59% vs. 40%; mean difference: 17.6 (95% confidence interval [CI] 11.1, 24.0)). Cost-effectiveness analyses showed that the model of care resulted in quality-adjusted life-years gained and cost-savings compared to usual care with mean INMB €2020 (95% CI 611, 3492) over 12 months. This study showed that implementing the model of care for OA in primary healthcare, improved quality of care and showed cost-effectiveness over 12 months compared to usual care. ClinicalTrials.gov NCT02333656.


DAPTA, a C-C chemokine receptor 5 (CCR5) antagonist attenuates immune aberrations by downregulating Th9/Th17 immune responses in BTBR T+ Itpr3tf/J mice.
Autism spectrum disorder (ASD) is a prevalent neurodevelopmental disorder characterized by deficits in social interaction, communication, and repetitive behaviors. BTBR T+ Itpr3tf/J (BTBR) mice, a preclinical autistic model featuring ASD symptoms as defined by social relations, was used in this study. We evaluated the potentially protective effect of D-Ala-peptide T-amide (DAPTA), a selective C-C chemokine receptor 5 (CCR5) antagonist, in BTBR mice. CCR5 is considered a potential therapeutic target in different neurodegenerative disorders. BTBR and C57 mice were intraperitoneally (i.p) treated with the DAPTA (0.01 mg/kg, i.p, once daily) for 7 days. We examined the effect of DAPTA by evaluating marble burying and administering repetitive behavior tests. We employed flow cytometry to assess the effect of DAPTA on CCR5+, CD4+CCR5+, CCR5+IL-6+, CCR5+IL-9+, CCR5+IL-17A+, CCR5+RORγT+, CCR5+IL-10+, and CCR5+Foxp3+ in spleen cells. We further explored the effects of DAPTA on IL-6, IL-9, IL-17A, RORγT, IL-10, and Foxp3 protein and mRNA expression levels in the brain tissues. DAPTA administration significantly decreased marble burying and repetitive behavior in BTBR mice. Additionally, DAPTA treatment inhibited CCR5+, CD4+CCR5+, CCR5+IL-6+, CCR5+IL-9+, CCR5+IL-17A+, CCR5+RORγT+, and upregulated CCR5+IL-10+, and CCR5+Foxp3+ production. We further observed that DAPTA downregulated IL-6, IL-9, IL-17A, and RORγT, and increased IL-10 and Foxp3 protein and mRNA expression. Therefore, our results suggest that DAPTA administration represents a potential treatment strategy for patients with ASD.


Infant sleep location: associated maternal and infant characteristics with sudden infant death syndrome prevention recommendations.
To identify factors associated with infant sleep location. Demographic information and infant care practices were assessed for 708 mothers of infants ages 0 to 8 months at Women, Infants and Children centers. Generalized linear latent mixed models were constructed for the outcome, sleeping arrangement last night (room-sharing without bed-sharing versus bed-sharing, and room-sharing without bed-sharing versus sleeping in separate rooms). Two-thirds of the mothers were African-American. A total of 48.6% mothers room-shared without bed-sharing, 32.5% bed-shared, and 18.9% slept in separate rooms. Compared with infants who slept in separate rooms, infants who room-shared without bed-sharing were more likely to be Hispanic (odds ratio [OR], 2.58, 95% CI 1.11-5.98) and younger (3.66- and 1.74-times more likely for infants 0-1 month old and 2-3 months old, respectively, as compared with older infants). Compared with infants who bed-shared, infants who room-shared without bed-sharing were more likely to be 0 to 1 month old (OR, 1.57; 95% CI, 1.05-2.35) and less likely to be African-American (OR, 0.43; 95% CI, 0.26-0.70) or have a teenage mother (OR, 0.37; 95% CI, 0.23-0.58). Approximately one-third of mothers and infants bed-share, despite increased risk of sudden infant death syndrome (SIDS). The factors associated with bed-sharing are also associated with SIDS, likely rendering infants with these characteristics at high risk for SIDS.


The relationship between serum ammonia level and neurologic complications in patients with acute glufosinate ammonium poisoning: A prospective observational study.
Glufosinate ammonium poisoning can cause neurological complications even after a symptom-free period. We prospectively investigated the predictors of neurologic complications in acute glufosinate ammonium poisoning and the change of serum ammonia level as a predictor of patient's presence and recovery of neurologic complication. This prospective observational study collected data from consecutive patients diagnosed with acute glufosinate ammonium poisoning between September 2014 and June 2016. Serum ammonia was serially measured. The patients were divided into two groups: the neurologic complication group and the nonneurologic complication group. We also defined 25 other insecticide- or herbicide-poisoned patients as controls. The neurologic complication group included 18 patients (72.0%). The latency period for neurologic complications was within 48-h postingestion. The peak ammonia level was statistically higher in the neurologic complication group than in the control group ( p < 0.001) and the nonneurologic complication groups ( p = 0.001). There was a statistical difference between the nonneurologic complication group and the neurologic complication group ( p = 0.0085) in terms of ingested amount. The peak ammonia was the only predictor for the development of neurologic complications (the optimal cutoff: 90 μg/dL). In patients with mental changes, the mean serum ammonia levels before and after recovery of the mental changes were statistically different ( p = 0.0019). In acute glufosinate ammonium poisoning, serial serum ammonia level measurements are needed and a serum peak ammonia level greater than 90 μg/dL is a predictor of neurologic complications. Also, it is important to treat the hyperammonemia in acute glufosinate ammonium poisoning.


Octreotide-induced hyperkalemia.
A 68-year-old woman was admitted with numerous injuries after being hit by an automobile. On day 16 of hospitalization octreotide was started for treatment of a pancreatic fistula, and within 2 days the patient's serum potassium increased. By the third and fourth days of octreotide therapy the level had increased to the point at which it required treatment. The patient was asymptomatic, however, and had normal renal function. Octreotide was discontinued on the fifth day of the course, with a subsequent decrease in serum potassium. Two days later, one dose of the drug was given; serial serum potassium levels were measured before and after the dose, and increased after the rechallenge. Octreotide was not administered for the remainder of the patient's hospitalization, and hyperkalemia did not recur.


Melinda - A custom search engine that provides access to locally-developed content using the HL7 Infobutton standard.
Healthcare organizations use care pathways to standardize care, but once developed, adoption rates often remain low. One challenge for usage concerns clinicians' difficulty in accessing guidance when it is most needed. Although the HL7 'Infobutton Standard' allows clinicians easier access to external references, access to locally-developed resources often requires clinicians to deviate from their normal electronic health record (EHR) workflow to use another application. To address this gap between internal and external resources, we reviewed the literature and existing practices at the University of Utah Health Care. We identify the requirements to meet the needs of a healthcare enterprise and clinicians, describe the design and development of a prototype to aggregate both internal and external resources from within or outside the EHR, and evaluated strengths and limitations of the prototype. The system is functional but not implemented in a live EHR environment. We suggest next steps and enhancements.


Development of a Flexible Artificial Lateral Line Canal System for Hydrodynamic Pressure Detection.
Surface mounted 'smart skin' can enhance the situational and environmental awareness of marine vehicles, which requires flexible, reliable, and light-weight hydrodynamic pressure sensors. Inspired by the lateral line canal system in fish, we developed an artificial lateral line (ALL) canal system by integrating cantilevered flow-sensing elements in a polydimethylsiloxane (PDMS) canal. Polypropylene and polyvinylidene fluoride (PVDF) layers were laminated together to form the cantilevered flow-sensing elements. Both the ALL canal system and its superficial counterpart were characterized using a dipole vibration source. Experimental results showed that the peak frequencies of both the canal and superficial sensors were approximately 110 Hz, which was estimated to be the resonance frequency of the cantilevered flow-sensing element. The proposed ALL canal system demonstrated high-pass filtering capabilities to attenuate low-frequency stimulus and a pressure gradient detection limit of approximately 11 Pa/m at a frequency of 115 ± 1 Hz. Because of its structural flexibility and noise immunity, the proposed ALL canal system shows significant potential for underwater robotics applications.


Distribution of sputum cellular phenotype in a large asthma cohort: predicting factors for eosinophilic vs neutrophilic inflammation.
Phenotyping asthma according to airway inflammation allows identification of responders to targeted therapy. Induced sputum is technically demanding. We aimed to identify predictors of sputum inflammatory phenotypes according to easily available clinical characteristics. This retrospective study was conducted in 508 asthmatics with successful sputum induction recruited from the University Asthma Clinic of Liege. Receiver-operating characteristic (ROC) curve and multiple logistic regression analysis were used to assess the relationship between sputum eosinophil or neutrophil count and a set of covariates. Equations predicting sputum eosinophils and neutrophils were then validated in an independent group of asthmatics. Eosinophilic (≥3%) and neutrophilic (≥76%) airway inflammation were observed in 46% and 18% of patients respectively. Predictors of sputum eosinophilia ≥3% were high blood eosinophils, FENO and IgE level and low FEV1/FVC. The derived equation was validated with a Cohen's kappa coefficient of 0.59 (p < 0.0001). ROC curves showed a cut-off value of 220/mm3 (AUC = 0.79, p < 0.0001) or 3% (AUC = 0.81, p < 0.0001) for blood eosinophils to identify sputum eosinophilia ≥3%. Independent predictors of sputum neutrophilia were advanced age and high FRC but not blood neutrophil count. Eosinophilic and paucigranulocytic asthma are the dominant inflammatory phenotypes. Blood eosinophils provide a practical alternative to predict sputum eosinophilia but sputum neutrophil count is poorly related to blood neutrophils.


Computer simulations of the diffusion of Na+ and Cl- ions across POPC lipid bilayer membranes.
We have carried out molecular dynamics simulations using NAMD to study the diffusivity of Na and Cl ions across a POPC lipid bilayer membrane. We show that an imbalance of positively and negatively charged ions on either side of the membrane leads to the diffusion of ions and water molecules. We considered the cases of both weak and very strong charge imbalance across the membrane. The diffusion coefficients of the ions have been determined from the mean square displacements of the particles as a function of time. We find that for strong electrochemical gradients, both the Na and Cl ions diffuse rapidly through pores in the membrane with diffusion coefficients up to ten times larger than in water. Rather surprisingly, we found that although the Na ions are the first to begin the permeation process due to the lower potential barrier that they experience compared to the Cl ions, the latter complete the permeation across the barrier more quickly due to their faster diffusion rates.


Sodium zinc hydroxide sulfite with a novel Zn3OH geometry.
The synthesis and crystal structure of a new sodium zinc hydroxide sulfite, Na[(Zn(OH)]3(SO3)2, with novel structural features and bonding geometry are reported. In Na[(Zn(OH)]3(SO3)2, Zn2+ and O2- alternate to form an interesting corrugated hexagonal sheet with rings consisting of (ZnO)3. Another rare occurrence is the bonding geometry of one-third of the O sites within the sheet. These O sites form a planar Zn3O unit, unlike other known compounds, in which Zn3O is pyramidal.


Presymptomatic testing for myotonic dystrophy by means of the linked DNA marker APOC2.
A family in which the gene for myotonic dystrophy is segregating was tested with a DNA probe to the apolipoprotein-C2 (APOC2) gene. The APOC2 gene is linked closely to the myotonic dystrophy (DM) gene and can be used as a marker to follow the transmission of the DM gene within families. Results from DNA-marker studies were combined with information from clinical, ophthalmological and electromyographic examinations, with age-dependent penetrance and a recombinant frequency of 4% between the genes for myotonic dystrophy and apolipoprotein C2 being taken into account. The chance that an individual had inherited the gene for myotonic dystrophy was determined by means of the computer program, LINKAGE. This analysis altered dramatically the assessed risk that certain individuals in the family were carrying the DM gene. For four individuals in the family, the previous risks of 50% for each member were reduced to 0.9%, 0.9% and 0.2%, respectively, in three cases and increased to 91% in the fourth case. The importance of a complete clinical evaluation of potential gene carriers who donate blood for a DNA-linkage study is stressed.


Cancer disclosure in Japan: historical comparisons, current practices.
Although Japanese physicians historically have not disclosed cancer diagnoses to patients, pressures upon physicians to disclose have increased in recent years. We questioned physicians practicing at a private medical hospital in rural Japan about their current approach to cancer disclosure. We compared their responses with responses of physicians in a 1991 study conducted in Japan, and two studies conducted in the United States, in 1961 and in 1977. Seventy-seven clinically active physicians with experience treating cancer patients responded (73% response rate). Forty percent of respondents reported usually telling patients of a cancer diagnosis, over three times more than the 13% who reported such a policy in Japan in 1991. Physicians were significantly more likely (P < 0.001) to make exceptions than physicians in the previous Japanese study, and physicians in the previous U.S. studies. Respondents reported considering more factors in deciding whether to tell than participants in the 1977 U.S. study. Factors predicting an increased probability of disclosure included age (in a hyperbolic relationship), improved prognosis, breast or cervical cancer, and social status and religion (by a minority of respondents). Increase in a substitute decision maker's age was predictive for physicians not to involve the person in decision-making. Most respondents reported their disclosure policy is based on clinical experience. More respondents indicated a likelihood of changing in the future than respondents in the previous U.S. studies. These data suggest Japanese physicians are moving away from a rigid policy of nondisclosure to a policy of selective disclosure, but they have not adopted universal cancer disclosure as practiced in many Western countries. Since young age and advanced age predicted self-reports of not disclosing cancer diagnoses, these data raise the question of whether nondisclosure by age groups represents age discrimination or compassionate medical care tailored to individual patient needs.


[HTLV-I/II in Switzerland: apropos of 4 case reports].
HTLV-I/II (human T-cell lymphotropic virus type I and II) infection is endemic in South Japan, Subsaharan Africa, the Caribbean and in regions of South America and of the United States. The infection almost always remains asymptomatic but is also associated with two distinct diseases: tropical spastic paraparesis (TSP), and adult T-cell leukemia (ATL). Although very rare in Europe, HTLV-I/II infection may occur in patients originating from endemic areas. We report the cases of four patients seropositive for HTLV-I/II, all of them living in Switzerland. Three of them originate from Zaire; the fourth is a Swiss female married to a Taiwanese. One patient has TSP; one has pyramidal signs along with systemic vasculitis-associated neuropathy; the third is HIV-position with stage III B infection, without symptoms of TSP or ATL; the last is entirely asymptomatic.


Preclinical characterization of mobocertinib highlights the putative therapeutic window of this novel EGFR inhibitor to EGFR exon 20 insertion mutations.
Epidermal growth factor receptor (EGFR) exon 20 insertion mutations account for 10% of all EGFR mutations and are mostly insensitive to approved EGFR-tyrosine kinase inhibitors (EGFR-TKIs). Novel EGFR-TKIs have been developed or repurposed for these mutants. A limited number of preclinical studies have detailed these EGFR-TKIs. We sought to use commercially available mobocertinib (TAK-788) to characterize the preclinical therapeutic window of this EGFR-TKI against EGFR mutations and to probe possible on-target mechanisms of resistance (EGFR-C797S). We used models of EGFR mutations to probe representative 1st, 2nd, 3rd generation, and in-development EGFR exon 20-active (poziotinib, mobocertinib) TKIs. We also introduced EGFR-C797S to these models to identify mechanisms of resistance. Cells driven by the most common EGFR exon 20 insertion mutations (A767_V769dupASV, D770_N771 insSVD, H773_V774insH and others) were inhibited by in-development EGFR TKIs at doses below those affecting EGFR-wildtype; albeit more common EGFR mutations (exon 19 deletions and L858R) were inhibited more readily by mobocertinib and poziotinib. Mobocertinib was able to inhibit phosphorylation of EGFR in multiple preclinical models. The presence of EGFR-C797S led to >200-fold resistance in proliferation assays probing mobocertinib and osimertinib. Review of clinical studies of mobocertinib disclosed responses that could be lasting. This is one of the initial reports to characterize the novel EGFR TKI mobocertinib and highlights its broad activity against EGFR mutants plus the therapeutic window to EGFR exon 20 insertion mutations; as well as EGFR-C797S as a possible mechanism of resistance. Further clinical development of mobocertinib merits continuation.


J-Type Heteroexciton Coupling Effect on an Asymmetric Donor-Acceptor-Donor-Type Fluorophore.
The novel donor-acceptor-donor (D-A-D)-type fluorophore with an asymmetric structure is reported. The twisted-induced charge transfer (TICT) luminescence was observed. The degree of charge transfer and radiative rate constant in the luminescence increased simultaneously with increase in orientational polarizability of solvents. In contrast to the numerous CT fluorophore researches, this behavior has never been previously observed. This characteristic behavior reveals the existence of an effective exciton coupling between the CT states in the donor-acceptor-donor-type fluorophore for the first time.


Plasma DCLK1 is a marker of hepatocellular carcinoma (HCC): Targeting DCLK1 prevents HCC tumor xenograft growth via a microRNA-dependent mechanism.
Tumor stem cell marker Doublecortin-like kinase1 (DCLK1) is upregulated in several solid tumors. The role of DCLK1 in hepatocellular carcinoma (HCC) is unclear. We immunostained tissues from human livers with HCC, cirrhosis controls (CC), and non-cirrhosis controls (NCC) for DCLK1. Western blot and ELISA analyses for DCLK1 were performed with stored plasma samples. We observed increased immunoreactive DCLK1 in epithelia and stroma in HCC and CCs compared with NCCs, and observed a marked increase in plasma DCLK1 from patients with HCC compared with CC and NCC. Analysis of the Cancer Genome Atlas' HCC dataset revealed that DCLK1 is overexpressed in HCC tumors relative to adjacent normal tissues. High DCLK1-expressing cells had more epithelial-mesenchymal transition (EMT). Various tumor suppressor miRNAs were also downregulated in HCC tumors. We evaluated the effects of DCLK1 knockdown on Huh7.5-derived tumor xenograft growth. This was associated with growth arrest and a marked downregulation of cMYC, and EMT transcription factors ZEB1, ZEB2, SNAIL, and SLUG via let-7a and miR-200 miRNA-dependent mechanisms. Furthermore, upregulation of miR-143/145, a corresponding decrease in pluripotency factors OCT4, NANOG, KLF4, and LIN28, and a reduction of let-7a, miR-143/145, and miR-200-specific luciferase activity was observed. These findings suggest that the detection of elevated plasma DCLK1 may provide a cost-effective, less invasive tool for confirmation of clinical signs of cirrhosis, and a potential companion diagnostic marker for patients with cirrhosis and HCC. Our results support evaluating DCLK1 as a biomarker for detection and as a therapeutic target for eradicating HCC.


[Rapamycin effect on senescence and autophagy processes in human cell lines].
Autophagy and senescence are both processes that firstly avoid tumor development through the inhibition of proliferation of damaged cells. However, autophagy does not imply cell death, because it is also a mechanism of cell survival under stress conditions. Concerning senescence, although these cells do not proliferate, they produce growth factors that contribute to the proliferative response of other cells. Rapamycin is an immunosupressor used in transplanted patients that inhibits the mTOR transduction signal pathway. This pathway is involved in the control of the energetic and nutritional state of the cell allowing protein synthesis and inhibiting autophagy when it is active. In this paper, the action of rapamycin over these processes was investigated and we found that a low concentration of this drug induces the senescence of a normal cell line, while a higher concentration induces autophagy of a transformed cell line. We have also determined that the oncogen RAC3 inhibits autophagy and that its expression is diminished by rapamycin. Therefore, our results contribute to a better understanding of the molecular mechanisms by which this drug is effective, given the relevance of rapamycin for potential tumor therapy.


High-resolution anoscopy: Unchartered territory for gastroenterologists?
High-resolution anoscopy (HRA) is a procedure where patients with an increased risk of anal cancer, like men who have sex with men, human immunodeficiency virus infected individuals, transplant patients and women with a history of lower genital tract neoplasia, with abnormal anal cytology results, are submitted to anal and perianal visualization under magnification. This will allow for a better detection of anal high-grade lesions that can be treated, in an effort to prevent anal cancer. Anal cancer screening follows the same principles that cervical cancer screening. During this procedure, an anoscope is inserted and a colposcope is used to examine systematically the squamocolumnar junction, the transformation zone and the perianal skin. Initially the observation is done with no staining and then with the application of acetic acid and Lugol's iodine solution, allowing for better lesion identification and characterization. Any suspicious lesion seen should be carefully evaluated and biopsied. Without HRA only a small percentage of suspicious lesions are identified. High-grade lesions that are detected can be ablated under HRA. This is a challenging exam to perform, with a long learning curve and the number of clinicians performing it is limited, although the growing number of patients that need to been screened. Specific equipment is required, with these patients ideally been followed by a multidisciplinary team, in a reference centre. HRA remains unfamiliar for many gastroenterologists.


Assembly and characterization of novel Alu inserts detected from next-generation sequencing data.
Repetitive elements generally, and Alu inserts specifically are a large contributor to the recent evolution of the human genome. By assembling the sequences of novel Alu inserts using their respective subfamily consensus sequences as references, we found an exponential decay in the Alu subfamily call enrichment with increased number of sequence variants (Pearson correlation [Formula: see text], [Formula: see text]). By mapping the sequences of these inserts to a human reference genome, we infer the reference Alu sources of a subset of the novel Alus, of which 85% were previously shown to be active. We also evaluate relationships between the loci of the novel inserts and their inferred sources.


Improved procedure for production of cytochalasin E and tremorgenic mycotoxins by Aspergillus clavatus.
This report describes an improved procedure for production of cytochalasin E and tremorgens by solid substrate, agitated fermentation of Aspergillus clavatus on pearled barley.


[Organization of collecting and analysis of the data on nutrition in epidemiological study of cardiovascular diseases].
The principles of collecting and analysis of information on nutrition in epidemiological study of cardiovascular diseases by means of electronic computers are described. A special program computes the consumption of 11 nutrients, the calorific value of the diet, and the calorie percentage of different nutrients with the exception of cholesterol. Certain coefficients are estimated, such as the ratio between the poly-unsaturated and the saturated fatty acids in the diet, and the ratio between the amount of starch and the total amount of refined and unrefined sugar, Well-trained technical personnel may be entrusted with the collection of the information, its coding, perforation, and feeding into the computer and with the obtainment of the data, which reduces the cost of the study considerably.


Hypolipidaemic efficacy of Capparis decidua fruit and shoot extracts in cholesterol fed rabbits.
High fat diet caused significant (8-fold) increase in serum total cholesterol in rabbits. Administration of C. decidua fruit extract (50% ethanolic) at the dose of 500 mg/kg body weight significantly reduced serum total cholesterol (61%), LDL cholesterol (71%), triglycerides (32%) and phospholipids (25%). Similarly C. decidua shoot extract lowered serum total cholesterol (48%), LDL cholesterol (57%), triglycerides (38%) and phospholipids (36%). The cholesterol content of aorta was decreased by 44 and 28% in fruit and shoot extract treatment respectively. The HDL to total cholesterol ratio and atherogenic index was significantly decreased in plant extract treated groups suggesting antiatherosclerotic nature of these plant extract. These results reveal the hypolipidaemic potential of C. decidua fruit and shoot.


Melatonin protects against the pathological cardiac hypertrophy induced by transverse aortic constriction through activating PGC-1β: In vivo and in vitro studies.
Melatonin, a circadian molecule secreted by the pineal gland, confers a protective role against cardiac hypertrophy induced by hyperthyroidism, chronic hypoxia, and isoproterenol. However, its role against pressure overload-induced cardiac hypertrophy and the underlying mechanisms remains elusive. In this study, we investigated the pharmacological effects of melatonin on pathological cardiac hypertrophy induced by transverse aortic constriction (TAC). Male C57BL/6 mice underwent TAC or sham surgery at day 0 and were then treated with melatonin (20 mg/kg/day, via drinking water) for 4 or 8 weeks. The 8-week survival rate following TAC surgery was significantly increased by melatonin. Melatonin treatment for 8 weeks markedly ameliorated cardiac hypertrophy. Compared with the TAC group, melatonin treatment for both 4 and 8 weeks reduced pulmonary congestion, upregulated the expression level of α-myosin heavy chain, downregulated the expression level of β-myosin heavy chain and atrial natriuretic peptide, and attenuated the degree of cardiac fibrosis. In addition, melatonin treatment slowed the deterioration of cardiac contractile function caused by pressure overload. These effects of melatonin were accompanied by a significant upregulation in the expression of peroxisome proliferator-activated receptor-gamma co-activator-1 beta (PGC-1β) and the inhibition of oxidative stress. In vitro studies showed that melatonin also protects against angiotensin II-induced cardiomyocyte hypertrophy and oxidative stress, which were largely abolished by knocking down the expression of PGC-1β using small interfering RNA. In summary, our results demonstrate that melatonin protects against pathological cardiac hypertrophy induced by pressure overload through activating PGC-1β.


Relevance of the light signaling machinery for cellulase expression in Trichoderma reesei (Hypocrea jecorina).
In nature, light is one of the most important environmental cues that fungi perceive and interpret. It is known not only to influence growth and conidiation, but also cellulase gene expression. We therefore studied the relevance of the main components of the light perception machinery of Trichoderma reesei (Hypocrea jecorina), ENV1, BLR1 and BLR2, for production of plant cell wall degrading enzymes in fermentations aimed at efficient biosynthesis of enzyme mixtures for biofuel production. Our results indicate that despite cultivation in mostly dark conditions, all three components show an influence on cellulase expression. While we found the performance of the enzyme mixture secreted by a deletion mutant in env1 to be enhanced, the higher cellulolytic activity observed for Δblr2 is mainly due to an increased secretion capacity of this strain. Δblr1 showed enhanced biomass accumulation, but due to its obviously lower secretion capacity still was the least efficient strain in this study. We conclude that with respect to regulation of plant cell wall degrading enzymes, the blue light regulator proteins are unlikely to act as a complex. Their regulatory influence on cellulase biosynthesis involves an alteration of protein secretion, which may be due to adjustment of transcription or posttranscriptional regulation of upstream factors. In contrast, the regulatory function of ENV1 seems to involve adjustment of enzyme proportions to environmental conditions.


Inflammation and perforation of a solitary diverticulum of the cecum. A report of 5 cases and literature review.
Solitary diverticulum of the cecum is a benign condition uncommon in the Western world, and with a higher incidence in Asian population. They are usually asymptomatic, and manifest clinically only with complications such as inflammation, perforation, or bleeding. They are a rare cause of acute abdominal pain, clinically similar to acute appendicitis, with tenderness in the right lower quadrant, fever, and leukocytosis. In spite of the information provided by ultrasonography or CT scans, a correct preoperative diagnosis is still difficult to reach, and is usually arrived at in the operating theater; differentiation from a neoplasm may be also sometimes complicated, and a wide surgical resection is usually required for such cases. We report on 5 cases of inflammation and perforation of a solitary cecal diverticulum, and perform a literature review.


Masked comparison of trypan blue stain and potassium hydroxide with calcofluor white stain in the microscopic examination of corneal scrapings for the diagnosis of microbial keratitis.
To evaluate the efficacy of trypan blue in direct microscopic examination of corneal scrapings in the diagnosis of non-viral microbial keratitis. In a prospective, interventional, masked study, 82 consecutive patients were investigated. Direct microscopic examination of the corneal scrapings involved three smears stained with potassium hydroxide with calcofluor white (KOH + CFW), Gram stain (not analyzed), and trypan blue stain and culture for bacteria, fungus, and Acanthamoeba. While KOH + CFW stained slides were examined under a fluorescence microscope, trypan blue-stained slides were examined by two microbiologists (masked to KOH + CFW and culture results) under normal light microscopy. Thirty samples were reexamined for interobserver and intraobserver variability. Out of 82 samples, fungal/fungus-like elements were seen in 48 (58.5%) in KOH + CFW and 38 (46.3%) in trypan blue stain. One microsporidial case, detected in KOH + CFW was negative in trypan blue stain (culture not done). Fungal growth was positive in 23 out of 81 (28.3%) cases cultured, single bacterial species in 18 (22.2%), Pythium insidiosum in three, mixed bacteria and fungus/Pythium in 7 (8.6%), Acanthamoeba in none and 30 (37.0%) samples were sterile. With culture as gold standard, the respective sensitivity (84.9%, 75.7%) and specificity (90.9%, 68.2%) of KOH + CFW stain and trypan blue stain were comparable (p = 0.16, P = 0.06). The interobserver linear weighted kappa score between the two microbiologists was 1.00 while it was 0.86 for intraobserver agreement. Trypan blue stain, an easily available dye to ophthalmologists, is highly efficacious in the diagnosis of fungal keratitis.


Human pancreatic beta-cell glucokinase: subcellular localization and glucose repression signalling function in the yeast cell.
Human GK(beta) (pancreatic beta-cell glucokinase) is the main glucose-phosphorylating enzyme in pancreatic beta-cells. It shares several structural, catalytic and regulatory properties with Hxk2 (hexokinase 2) from Saccharomyces cerevisiae. In fact, it has been previously described that expression of GK(beta) in yeast could replace Hxk2 in the glucose signalling pathway of S. cerevisiae. In the present study we report that GK(beta) exerts its regulatory role by association with the yeast transcriptional repressor Mig1 (multicopy inhibitor of GAL gene expression 1); the presence of Mig1 allows GK(beta) to bind to the SUC2 (sucrose fermentation 2) promoter, helping in this way in the maintenance of the repression of the SUC2 gene under high-glucose conditions. Since a similar mechanism has been described for the yeast Hxk2, the findings of the present study suggest that the function of the regulatory domain present in these two proteins has been conserved throughout evolution. In addition, we report that GK(beta) is enriched in the yeast nucleus of high-glucose growing cells, whereas it shows a mitochondrial localization upon removal of the sugar. However, GK(beta) does not exit the nucleus in the absence of Mig1, suggesting that Mig1 regulates the nuclear exit of GK(beta) under low-glucose conditions. We also report that binding of GK(beta) to Mig1 allows the latter protein to be located at the mitochondrial network under low-glucose conditions.


The Master Protocol Concept.
During the past decade, biomedical technologies have undergone an explosive evolution-from the publication of the first complete human genome in 2003, after more than a decade of effort and at a cost of hundreds of millions of dollars-to the present time, where a complete genomic sequence can be available in less than a day and at a small fraction of the cost of the original sequence. The widespread availability of next-generation genomic sequencing has opened the door to the development of precision oncology. The need to test multiple new targeted agents both alone and in combination with other targeted therapies, as well as classic cytotoxic agents, demands the development of novel therapeutic platforms (particularly Master Protocols) capable of efficiently and effectively testing multiple targeted agents or targeted therapeutic strategies in relatively small patient subpopulations. Here, we describe the Master Protocol concept, with a focus on the expected gains and complexities of the use of this design. An overview of Master Protocols currently active or in development is provided along with a more extensive discussion of the Lung Master Protocol (Lung-MAP study).


Conformational changes in the extrinsic manganese stabilizing protein can occur upon binding to the photosystem II reaction center: an isotope editing and FT-IR study.
Photosystem II catalyzes the light-driven oxidation of water and reduction of plastoquinone in oxygenic photosynthesis. The manganese stabilizing protein (MSP) of photosystem II is an extrinsic subunit that plays an important role in catalytic activity. This subunit can be extracted and re-bound to the photosystem II reaction center. Extraction is associated with decreased stability of manganese binding by the enzyme and by loss in high rates of oxygen evolution activity; reconstitution reverses these phenomena. Since little is known about the assembly of complex membrane proteins, we have employed isotope editing and vibrational spectroscopy to obtain information about any changes in secondary structure that occur in MSP upon functional reconstitution to photosystem II. The spectroscopic data obtained are consistent with substantial changes in conformation when MSP binds to photosystem II; approximately 30-40% of the peptide backbone undergoes a change in secondary structure. These conclusions were reached by comparing different aliquots, before and after binding, of the same 13[C]MSP sample. Analysis of amide I band line shapes through Fourier deconvolution and nonlinear regression suggests that binding of MSP to photosystem II is associated with a decrease in random structure and an increase in beta-sheet content. We conclude that binding of MSP to the reaction center can induce folding of MSP. Our results also indicate that, in solution, MSP can sample a variety of conformational states, which differ in hydrogen bonding of the peptide backbone.


Chemical- and irradiation-induced mutants of indica rice IR64 for forward and reverse genetics.
IR64, the most widely grown indica rice in South and Southeast Asia, possesses many positive agronomic characteristics (e.g., wide adaptability, high yield potential, tolerance to multiple diseases and pests, and good eating quality,) that make it an ideal genotype for identifying mutational changes in traits of agronomic importance. We have produced a large collection of chemical and irradiation-induced IR64 mutants with different genetic lesions that are amenable to both forward and reverse genetics. About 60,000 IR64 mutants have been generated by mutagenesis using chemicals (diepoxybutane and ethylmethanesulfonate) and irradiation (fast neutron and gamma ray). More than 38,000 independent lines have been advanced to M4 generation enabling evaluation of quantitative traits by replicated trials. Morphological variations at vegetative and reproductive stages, including plant architecture, growth habit, pigmentation and various physiological characters, are commonly observed in the four mutagenized populations. Conditional mutants such as gain or loss of resistance to blast, bacterial blight, and tungro disease have been identified at frequencies ranging from 0.01% to 0.1%. Results from pilot experiments indicate that the mutant collections are suitable for reverse genetics through PCR-detection of deletions and TILLING. Furthermore, deletions can be detected using oligomer chips suggesting a general technique to pinpoint deletions when genome-wide oligomer chips are broadly available. M4 mutant seeds are available for users for screening of altered response to multiple stresses. So far, more than 15,000 mutant lines have been distributed. To facilitate broad usage of the mutants, a mutant database has been constructed in the International Rice Information System (IRIS; http: //www.iris.irri.org) to document the phenotypes and gene function discovered by users.