FERM domain containing protein 7 (FRMD7) upregulates the expression of neuronal cytoskeletal proteins and promotes neurite outgrowth in Neuro-2a cells.
Mutations of the FERM domain containing protein 7 gene (FRMD7) are associated with X-linked idiopathic congenital nystagmus. Previous studies have shown that FRMD7 plays an important role in neuronal development and is involved in the regulation of F-actin. However, its specific mechanism of action remains undetermined. Our study used quantitative real-time PCR to assess the levels of neuron-specific genes in a mouse neuroblastoma cell line (Neuro-2a) after transfection with a full-length coding transcript of FRMD7 or a blank control vector. F-actin was detected by rhodamine-phalloidin staining. Neurite number and length were assessed by a confocal laser scanning microscope. We demonstrated that FRMD7 can promote neurite outgrowth following retinoic acid-induced differentiation in Neuro-2a cells. Neurites were significantly longer in cells transfected with FRMD7, but there was no difference in cell numbers. The mRNA expression of neuron cytoskeletal-related genes (microtubule-associated protein 2 [Mtap2], neurofilament-L and M [NF-L and NF-M] and the microtubule-associated protein tau [MAPT]) were significantly increased compared to controls. Other genes (NF-H, MAPT, neuron-specific class III beta-tubulin (Tuj-1), nestin, and growth-associated protein-43 [GAP-43]) were not obviously altered by FRMD7 overexpression. Taken together, our data suggest that FRMD7 promotes the extension of neurites and may be involved in regulating the movement of cytoskeletal proteins, which influences not only F-actin, but also NF and microtubule dynamics.


Effect of aquatic versus land motor dual task training on balance and gait of patients with chronic stroke: A randomized controlled trial.
Patients with stroke are more likely to be at risk of falling, which lead to limitation in their abilities to perform daily living activities and participation in society. Falling is due to an increase in postural sway and a decline in gait ability. Exercise in water helps them to safely and comfortably improve their motor ability by providing low-risk exercise environments. This study was conducted to compare the effect of aquatic versus land motor dual task training on balance and gait of patients with chronic stroke. Fifty patients with chronic stroke of both sexes aging from 45 to 55 years were randomly assigned to aquatic or land group. Both groups received the same motor dual task training either in water or on land for 45 minutes, 3 days a week for six weeks. Measurement of the dynamic balance indices assessed using Biodex Balance System as well as kinematic gait parameters using Biodex Gait Trainer was performed before and after the intervention. There was a significant improvement in all outcome variables post treatment compared with that pre-treatment in both groups (P < 0.05). There was a significant improvement in patients who received the motor dual task training in water compared with patients treated on the land in overall stability index (P = 0.02), anteroposterior stability index (P = 0.03), mediolateral stability index (P = 0.002), walking speed (P = 0.01), step length of affected limb (P = 0.03), step length of non-affected limb (P = 0.01), and time of support on the affected limb (P = 0.002). Aquatic motor dual task training is more effective in improving balance and gait abilities of patients with chronic stroke than land motor dual task training.


A combined imagistic and morphological approach of lung tumors: study on 64 cases.
Lung cancer is currently one of the major health problems, being considered one of the most common causes of death by cancer worldwide. That is why establishing as early as possible a diagnosis in order to start an appropriate therapy still a challenge. The studied group consisted of 64 patients who were investigated following the next algorithm: chest X-ray, bronchoscopy, guided bronchial brushing and/or bronchoalveolar lavage followed by cytological examination and if possible, endobronchial biopsy followed by histopathological examination. Patients were usually men, aged over 60 years coming from an urban area, smokers and with symptoms evoking the presence of neoplasia. Tumors presented as large tumoral masses, placed centrally and with obvious local spread, protruding or pushing intraluminally with subsequent bronchial stenosis, usually complicated with ulceration and bleeding, which proved to be, when histopathological examination was possible, firstly squamous carcinomas and then small cell carcinomas. The imagistic investigation in conjunction with the cytological evaluation can establish the diagnosis of malignancy of centrally located lung tumors in almost 80% of cases. The diagnosis can be improved by increasing the number of biopsy sampling and/or by the immunohistochemical marking of the cytological samples.


Value for money of changing healthcare services? Economic evaluation of quality improvement.
There are many instances of perceived or real inefficiencies in health service delivery. Both healthcare providers and policy makers need to know the impact and cost of applying strategies to change the behaviour of individuals or organisations. Quality improvement or implementation research is concerned with evaluating the methods of behavioural change. Addressing inefficiencies in healthcare services raises a series of issues, beginning with how inefficiency itself should be defined. The basic concepts of cost analysis and economic evaluations are explained and a model for working through the economic issues of quality improvement is discussed. This model combines the costs and benefits of corrected inefficiency with the costs and degree of behavioural change achieved by a quality improvement method in the policy maker's locality. It shows why it may not always be cost effective for policy makers to address suboptimal behaviour. Both the interpretation of quality improvement research findings and their local application need careful consideration. The limited availability of applicable quality improvement research may make it difficult to provide robust advice on the value for money of many behavioural quality improvement strategies.


Exploring the anticancer effects of standardized extracts of poplar-type propolis: In vitro cytotoxicity toward cancer and normal cell lines.
Propolis was shown to exert antimicrobial, antioxidant, anti-inflammatory, and anticancer activities. Its composition is influenced by seasonal, climatic and phytogeographic conditions. Further variability derives from the extraction methods. Multi Dynamic Extraction Method (MED) has been recently proposed to improve extracts reproducibility. Here, the cytotoxic/anticancer activity of three MED extracts of poplar-type propolis was assayed on human promyelocytic leukaemia HL60, human monocytic leukaemia THP-1, human osteosarcoma MG63, murine fibroblast L929 and human mesenchymal cells (hMSCs). As far as we are aware of, MG63 cells have never been challenged with propolis before, while few studies have so far addressed the effects of propolis on non-tumor cell lines. Consistent results were observed for all propolis preparations. The extracts turned out mildly cytotoxic toward cancer cells, in particular osteosarcoma cells (IC50: 81.9-86.7 µg/ml). Nonetheless, cytotoxicity was observed also in non-tumor L929 cells, with an even lower IC50. hMSCs demonstrated the lowest sensitivity to propolis (IC50: 258.3-287.2 µg/ml). In THP-1 cells, extracts were found to stimulate apoptosis caspase 3/7 activity. The IC50 values observed with osteosarcoma and leukaemia cells do not support a relevant cytotoxicity (as the figures abundantly exceeded 30 µg/ml), despites some selective activity exhibited with HL60 cells. The results confirm the validity of the extraction method, emphasizing the need to assess the selectivity of the interaction with cancer cells when screening for anticancer-drug candidates.


Diversity of fecal coliforms and their antimicrobial resistance patterns in wastewater treatment model plant.
The occurrence of resistance patterns among wastewater fecal coliforms was determined in the study. Susceptibility of the isolates was tested against 19 antimicrobial agents: aminoglycosides, aztreonam, carbapenems, cephalosporines, beta-lactam/beta-lactamase inhibitors, penicillines, tetracycline, trimethoprim/sulfamethoxazole, and fluoroquinolones. Additionally the removal of resistant isolates was evaluated in the laboratory-scale wastewater treatment model plant (M-WWTP), continuously supplied with the wastewater obtained from the full-scale WWTP. Number of fecal coliforms in raw (after mechanical treatment) and treated wastewater, as well as in aerobic chamber effluent was determined using selective medium. The selected strains were identified and examined for antibiotic resistance using Phoenix Automated Microbiology System (BD Biosciences, USA). The strains were identified as Escherichia coli (n=222), Klebsiella pneumoniae ssp. ozaenae (n=9), and Pantoea agglomerans (n=1). The isolate of P. agglomerans as well as 48% of E. coli isolates were sensitive to all antimicrobials tested. The most frequent resistance patterns were found for ampicillin: 100% of K. pneumoniae ssp. ozaenae and 41% of E. coli isolates. Among E. coli isolates 12% was regarded as multiple antimicrobial resistant (MAR). In the studied M-WWTP, the applied activated sludge processes reduced considerably the number of fecal coliforms, but increased the ratio of antimicrobial-resistant E. coli isolates to sensitive ones, especially among strains with MAR patterns.


Overweight induced by high-fat diet delays rat cutaneous wound healing.
Prolonged wound healing is a complication that contributes to morbidity and mortality. Overweight people regularly undergo surgery and trauma, and often develop chronic wounds, but the effects of the adipose tissue excess on cutaneous wound healing are not well understood. This study tested the hypothesis that overweight induced by a high-fat diet impairs rat cutaneous wound healing. Male Wistar rats were fed with either a high-fat or a standard (control) diet. After 15 weeks, an excisional lesion was done and the animals were killed 21 d later. Wound contraction and re-epithelialization, blood pressure, glucose and retroperitoneal fat were evaluated. After killing, lesion and adjacent normal skin were formol-fixed and paraffin-embedded. Inflammatory infiltrate, myofibroblasts, collagen fibres and cellular proliferation were analysed and blood vessels were evaluated using stereological methods. There was no difference in blood pressure and glucose, but retroperitoneal fat increased in the high-fat diet group. Animals fed with the high-fat diet presented delayed wound contraction and re-epithelialization. It was found that 21 d after wounding, overweight induced by a high-fat diet increased the inflammatory infiltrate and delayed myofibroblastic differentiation, collagen deposition, epithelial and connective tissue cell proliferation, and angiogenesis. These findings support the hypothesis that a high-fat diet exerts negative effects on rat cutaneous wound healing, due mainly to the prolongation of the inflammatory phase.


Is neoadjuvant chemotherapy followed by radical surgery more effective than radiation therapy for stage IIB cervical cancer?
The primary objective of the study was to compare the survival rate of patients who had received neoadjuvant chemotherapy with that of patients who had received radiation therapy for stage IIB cervical cancer. The secondary objective was to analyze the effect of neoadjuvant chemotherapy on pathological prognostic factors. We retrospectively reviewed the medical records of patients who had received therapy for stage IIB cervical cancer. Based on the primary therapy, 192 patients were divided into 2 groups; patients in the neoadjuvant chemotherapy group (n =103) underwent a type III radical hysterectomy after completion of the neoadjuvant chemotherapy. Patients in the other group (n = 89) were treated with radiation alone or a combination of chemotherapy and radiotherapy. After neoadjuvant chemotherapy, the level of squamous cell carcinoma antigen, tumor size, lymph node involvement, and parametrium involvement were significantly decreased. However, 90.3% of the patients who had received neoadjuvant chemotherapy needed to have adjuvant therapy after radical surgery because of poor pathological prognostic factors. The rate of disease-free survival did not differ significantly between the 2 groups. However, the overall survival rate was significantly lower in the neoadjuvant chemotherapy group for patients who were 60 years or older (P = 0.03). The rates of disease-free survival and overall survival for patients who had a good (complete or partial) response to the neoadjuvant chemotherapy were not significantly higher than the rates for patients in the radiation therapy group. Although neoadjuvant chemotherapy improved pathological prognostic factors in patients with stage IIB cervical cancer, it was not sufficiently effective to decrease adjuvant therapy. Neoadjuvant chemotherapy also did not improve the rate of patient survival compared to the rate of patient survival in the radiation therapy group.


Improving the Ability to Write Persuasive Texts in a Boy with Autism Spectrum Disorder: Outcomes of an Intervention.
In this paper, we describe an intervention implemented to assist a 13.2-year-old boy with Autism Spectrum Disorder, G, without intellectual disability, aimed at improving his ability to compose persuasive texts. There was an initial assessment (baseline), an intermediate assessment after two weeks, a six-session intervention phase, and a post-intervention assessment. Our intervention applied two procedures. The first aimed at enhancing general composition abilities in terms of picking (P) ideas, organizing (O) notes, and writing (W) them down (POW), while the second specified the steps to write a persuasive text addressing a possible reader: a topic sentence (T), reasons (R), an explanation (E) for the reasons and the end of the sentence (E) (TREE). These procedures were termed POW + TREE. To analyze G's texts, three types of measures were used by two raters at baseline, intermediate and post-test time: (a) the presence of the TREE components; (b) the quality of the reasons and explanations for the reasons; (c) the number of mental state terms. All these measures showed relevant quantitative improvements, as well as qualitative changes. In addition, when G's performance at the end of the intervention was compared to that of typically developing controls, no statistical difference appeared. The results are discussed in light of the potentialities offered by the type of intervention described here.


High toxoplasmosis seroprevalence among young pregnant women in Medea, Algeria.
Toxoplasmosis is a parasitic disease with worldwide distribution and a major public health problem. In fact, the zoonotic pathogen Toxoplasma gondii infects humans and almost all warm-blooded animals. One of the most common sources of human T. gondii infection is the ingestion of tissue cysts in raw or undercooked meat. The current epidemiological study was carried out in Medea (Algeria), from January to June 2017, using Enzyme Linked Immunosorbent assay (ELISA). Antibodies against T. gondii were determined in sera of more than one thousand pregnant women. The different age groups were: 15-20, 20-25, 25-30, 30-35, 35-40 and 40-45. Among 1,012 pregnant women analyzed, 252 (25%) were found seropositive, 49 among them showed a IgG+/IgM+ profile, and 760 (75%) were seronegative exhibiting IgG-/IgM-. Toxoplasmosis prevalence's were 44.5% and 39.5% in 15-20 and 20-25 age categories, respectively. A long-term prevention program is needed and strategies should be focused on the serodiagnosis of toxoplasmosis, especially in pregnant women. Since the consumption of raw or undercooked meat is not a part of the culinary culture in that area of study, cat exposure remains the principal factor of human infection.


MRSA hotspots.
The UK appears in the middle of the European MRSA league table between Scandinavia and Romania, but the statistics are not all they seem.


[Illness due to work or to an overactive thyroid?].
An incapacity to work brought about by stress was diagnosed in two women aged 40 and 38. They were both in conflict situations at work and had psychological problems. They stopped working but remained tired and irritable. After being unfit for work for more than a year they were diagnosed with hyperthyroidism which was treated appropriately. They both recovered and resumed work. Hyperthyroidism can impair physical and mental functioning. Presenting symptoms may be confusing, specifically if work-related factors appear to be the obvious implication. In The Netherlands, employees who become ill, consult their general practitioner or a medical specialist, who are concerned with diagnosis and treatment. They also report to their employer's occupational-health department, where the physician focuses on work-related factors that may impair health and on assessment of disability. Communication between all parties should be optimal.


Enantioselective and diastereoselective hydroxylation of bufuralol. Absolute configuration of the 7-(1-hydroxyethyl)-2-[1-hydroxy-2-(tert-butylamino)ethyl]benzofurans, the benzylic hydroxylation metabolites.
Asymmetric synthesis of the diastereomeric 7-(1-hydroxyethyl)-2-[1-hydroxy-2-(tert-butylamino)ethyl]benzofurans (2), the benzylic hydroxylation metabolites of bufuralol (1), is described, and the absolute configurations of these diastereomers are assigned. 1"-Oxobufuralol (3) was reduced with a complex of (2S)-(-)-2-amino-3-methyl-1,1-diphenylbutan-1-ol and borane, yielding 2, which had a 95:5 ratio of the possible 1"R and 1"S isomers as determined by HPLC. Separation of the resulting diastereomers was facilitated by derivatization with the enantiomers of 1-phenethyl isocyanate (PEIC). The absolute configurations 1'S,1"R and 1'R,1"R were assigned to the diastereomers formed in excess, 2c and 2b, on the basis of the known stereochemistry of reduction of closely related alkyl phenyl ketones to R alcohols by using this chiral borane reagent. The circular dichroism spectra of the four isomeric benzylic alcohols were in agreement with these assignments. In the presence of the rat liver microsomal fraction, benzylic hydroxylation of bufuralol was significantly product stereoselective favoring formation of diastereomers with the 1"R absolute stereochemistry at the new chiral center in products from (1'R)-1 by a ratio of 4.5:1 [(1'R,1"R)-2:(1'R,1"S)-2] and by nearly 8:1 [(1'S,1"R)-2:(1'S,1"S)-2] from (1'S)-1. (1'R)-Bufuralol was more rapidly hydroxylated than was (1'S)-1, by about 3-fold. In the presence of human liver microsomes, (1'R)-bufuralol was also more rapidly hydroxylated than was (1'S)-1, by ca. 2.5-fold. However, product stereoselectivity from the 1'R enantiomer was reversed from that observed in the rat liver microsomal oxidation, with more (1"S)-carbinol being formed than 1"R isomer by nearly 4-fold. From (1'S)-1, about equal amounts of the two possible hydroxybufuralol diastereomers were formed. The results from the human liver microsomal studies are consistent with observed enantioselectivity of hydroxylation of bufuralol in vivo in humans.


Hodgkin's lymphoma mortality in the Americas, 1997-2008: achievements and persistent inadequacies.
Although therapeutic advancements have made Hodgkin's lymphoma (HL) a largely curable disease, trends in HL mortality have been variable across countries. To provide updated information on HL mortality in the Americas, overall and 20-44 years age-standardized (world population) mortality rates from HL were derived for the 12 Latin American countries providing valid data to the World Health Organization database and with more than two million of inhabitants. For comparative purpose, data for the United States and Canada were also presented. Trends in mortality over the 1997 to 2008 period are based on joinpoint regression analysis. Declines in HL mortality were registered in all Latin American countries except in Venezuela. In most recent years, HL mortality had fallen to about 0.3/100,000 men and 0.2/100,000 women in Argentina, Brazil, Chile, Colombia, Ecuador and Guatemala, that is, to values similar to North America. Despite some declines, rates remained high in Cuba (1/100,000 men and 0.7/100,000 women), Costa Rica and Mexico as well as in Venezuela (between 0.5 and 0.6/100,000 men and between 0.3 and 0.5/100,000 women). In young adults, trends were more favorable in all Latin American countries except Cuba, whose rates remained exceedingly high (0.8/100,000 men and 0.6/100,000 women). Thus, appreciable declines in HL mortality were observed in most Latin America over the last decade, and several major countries reached values comparable to North America. Substantial excess mortality was still observed in Cuba, Costa Rica, Mexico and Venezuela, calling for urgent interventions to improve HL management in these countries.


A New Filter Approach Based on Effective Ranges for Classification of Gene Expression Data.
Over the years, many studies have been carried out to reduce and eliminate the effects of diseases on human health. Gene expression data sets play a critical role in diagnosing and treating diseases. These data sets consist of thousands of genes and a small number of sample sizes. This situation creates the curse of dimensionality and it becomes problematic to analyze such data sets. One of the most effective strategies to solve this problem is feature selection methods. Feature selection is a preprocessing step to improve classification performance by selecting the most relevant and informative features while increasing the accuracy of classification. In this article, we propose a new statistically based filter method for the feature selection approach named Effective Range-based Feature Selection Algorithm (FSAER). As an extension of the previous Effective Range based Gene Selection (ERGS) and Improved Feature Selection based on Effective Range (IFSER) algorithms, our novel method includes the advantages of both methods while taking into account the disjoint area. To illustrate the efficacy of the proposed algorithm, the experiments have been conducted on six benchmark gene expression data sets. The results of the FSAER and the other filter methods have been compared in terms of classification accuracies to demonstrate the effectiveness of the proposed method. For classification methods, support vector machines, naive Bayes classifier, and k-nearest neighbor algorithms have been used.


Prostatic carcinoma: treatment of liver metastases with intravenous diethylstilbestrol diphosphate.
A patient with Stage B adenocarcinoma of prostate treated with radical prostatectomy and interstitial radioactive gold presented ten years later with liver metastases without evidence of local recurrence. This patient was treated only with massive doses of intravenous diethylstilbestrol diphosphate, with regression of metastases and marked decline of the acid and alkaline phosphatase levels.


Modulation of quorum sensing-controlled virulence factors in Chromobacterium violaceum by selective amino acids.
Bacterial pathogenesis regulation requires N-acyl homoserine lactone (AHL)-based quorum sensing (QS). The main objective of this study was to assess the anti-QS and anti-biofilm potential of five different amino acids, namely serine, aspartic acid, lysine, leucine and tryptophan. The selected amino acids were assessed for their ability to inhibit QS activity via such mechanisms as exopolysaccharide (EPS) production, biofilm formation, pigment production in Chromobacterium violaceum and swarming motility. At 0.684 mM concentration, lysine inhibited the biofilm formation by 16% at 24 h, chitinolytic activity by 88.3% at 24 h and EPS production by 12.5% at 24 h. It also exhibited inhibition of swarming motility in C. violaceum. Confocal laser scanning microscopy revealed a decrease in the average thickness of the biofilms when treated with lysine. Modulation in the expression of cvi I and cvi A was observed when treated with all the amino acids, with the greatest reduction observed with lysine. Our results conclude that the amino acid lysine showed anti-QS and significant anti-biofilm activities; it could be further exploited as a main constituent in the synthesis of peptide/protein, and testing the same for treatment of bacterial infections, eventually reducing the utilization of conventional antibiotics.


Dissociation kinetics and equilibrium binding properties of polyene antibiotic complexes with phosphatidylcholine/sterol vesicles.
The interactions of sonicated vesicles with the polyene antibiotics amphotericin B, candicidin, mediocidin , and a water-soluble, guanidine derivative of amphotericin B were examined by UV-visible spectroscopy at concentrations below which the polyenes become self-associated. The association constants, Kapp, and the numbers of binding sites per sterol or phospholipid molecule (n) were determined at 30 degrees C and pH 7.4. A single class of binding sites was found, with no evidence of cooperativity. For the binding of mediocidin , amphotericin B, and the guanidine derivative with phosphatidylcholine (PC), PC/cholesterol, and PC/ergosterol vesicles, Kapp was in the range of (1.0-3.0) X 10(6) M-1; Kapp was higher for candicidin-vesicle interaction, reaching 9.0 X 10(6) M-1 with PC/ergosterol vesicles. Binding of the guanidine derivative of amphotericin B to PC vesicles lacking sterol was extensive (n = 0.46); since the other polyenes, which have low aqueous solubilities, had n less than 0.05, positive charges in the mycosamine moiety appear to enhance the extent of polyene antibiotic interaction with the glycerophospholipid head group. Higher values of n (and, therefore, of nKapp ) were found with sterol-containing than with sterol-free vesicles, suggestive of penetration of the polyenes toward the interior of the bilayer when sterol is present. For binding to PC/sterol vesicles, nKapp followed the order of candicidin greater than guanidine derivative of amphotericin B greater than amphotericin B much greater than mediocidin . The values of n and nKapp were appreciably higher for amphotericin B-ergosterol than for amphotericin B-cholesterol interaction in vesicles.(ABSTRACT TRUNCATED AT 250 WORDS)


Causes and frequency of tocophobia - own experiences.
The phenomenon of tocophobia (pathological fear of labor) has not been fully explored. Currently, there are no diagnostic criteria that would enable its detection or make it recognizable as a disease entity. The aim of study was to determine the degree of anxiety/tocophobia as well as to learn about and analyze the causes of this phenomenon in Polish pregnant women. The prospective study included 120 pregnant women in the third trimester of gestation from February to May 2016. The study was performed with the use of a standardized and revised version of the Labor Anxi-ety Questionnaire (KLP II) designed by Putyński and Paciorek (1997) as well as a proprietary interview questionnaire and structured data documentation form. In 6.7% of the subjects, the level of labor anxiety was very high (> 18 in KLP II). It was the highest in women over 30 years of age (Pearson's chi squared test = 0.00422; p < 0.05). It has been shown that successive childbirths have an im-pact on the degree of anxiety (p = 0.04217).The highest level of anxiety was noted in primiparous women. In 85% of the subjects, anxiety was caused by fear of labor pain. 56.7% of the tested women did not use any professional help in the preparation for childbirth and motherhood. Primiparas and women over 30 years of age experience very high levels of fear significantly more frequently. The lack of proper preparation for childbirth determines the occurrence of tocophobia. Ante-natal classes and prenatal education based on standards of obstetric care should be promoted in order to reduce or eliminate fear of natural labor.


Mitochondrial dysfunction enhances Gal4-dependent transcription.
Mitochondrial dysfunction has been shown to elicit broad effects on nuclear gene expression. We show here that transcription dependent on the prototypical acidic activator Gal4 is responsive to mitochondrial dysfunction. In cells with no mitochondrial DNA, Gal4-dependent gene expression is elevated. A minimal Gal4 activator containing the DNA binding and activation domain is sufficient for this response. Transcription dependent on a fusion of Gal4 to a heterologous DNA binding domain is similarly elevated in a mitochondrial mutant. Analysis of different Gal4-dependent promoters and gel mobility shift assays suggest that the effect of mitochondrial dysfunction on Gal4 activity is related to increased DNA binding to the cognate Gal4 element. Given that fermentation is the only means to obtain energy in respiratory deficient cells, it is possible that higher Gal4 activity in cells with dysfunctional mitochondria works to promote more efficient fermentation of galactose.


Near-instant surface-selective fluorogenic protein quantification using sulfonated triarylmethane dyes and fluorogen activating proteins.
Agonist-promoted G-protein coupled receptor (GPCR) endocytosis and recycling plays an important role in many signaling events in the cell. However, the approaches that allow fast and quantitative analysis of such processes still remain limited. Here we report an improved labeling approach based on the genetic fusion of a fluorogen activating protein (FAP) to a GPCR and binding of a sulfonated analog of the malachite green (MG) fluorogen to rapidly and selectively label cell surface receptors. Fluorescence microscopy and flow cytometry demonstrate that this dye does not cross the plasma membrane, binds with high affinity to a dL5** FAP-GPCR fusion construct, activating tagged surface receptors within seconds of addition. The ability to rapidly and selectively label cell surface receptors with a fluorogenic genetically encoded tag allows quantitative imaging and analysis of highly dynamic processes like receptor endocytosis and recycling.


From Variation of Influenza Viral Proteins to Vaccine Development.
Recurrent influenza epidemics and occasional pandemics are one of the most important global public health concerns and are major causes of human morbidity and mortality. Influenza viruses can evolve through antigen drift and shift to overcome the barriers of human immunity, leading to host adaption and transmission. Mechanisms underlying this viral evolution are gradually being elucidated. Vaccination is an effective method for the prevention of influenza virus infection. However, the emergence of novel viruses, including the 2009 pandemic influenza A (H1N1), the avian influenza A virus (H7N9), and the highly pathogenic avian influenza A virus (HPAI H5N1), that have infected human populations frequently in recent years reveals the tremendous challenges to the current influenza vaccine strategy. A better vaccine that provides protection against a wide spectrum of various influenza viruses and long-lasting immunity is urgently required. Here, we review the evolutionary changes of several important influenza proteins and the influence of these changes on viral antigenicity, host adaption, and viral pathogenicity. Furthermore, we discuss the development of a potent universal influenza vaccine based on this knowledge.


Evidence-based clinical practice guidelines for bladder cancer (summary - JUA 2009 Edition).
In Japan, until now, the treatment of bladder cancer has been based on guidelines from overseas. The problem with this practice is that the options recommended in overseas guidelines are not necessarily suitable for Japanese clinical practice. A relatively large number of clinical trials have been conducted in Japan in the field of bladder cancer, and the Japanese Urological Association (JUA) considered it appropriate to formulate their own guidelines. These Guidelines present an overview of bladder cancer at each clinical stage, followed by clinical questions that address problems frequently faced in everyday clinical practice. In this English translation of a shortened version of the original Guidelines, we have abridged each overview, summarized each clinical question and its answer, and only included the references we considered of particular importance.


Complications in the tissue integrated prostheses components: clinical and mechanical evaluation.
Cases demonstrating complications in osseointegrated implant supported prostheses are described. These complications occurred in the prostheses framework or veneer, in the fixation screw, in the abutment post or screw as well as in the implant body. The two main reasons for the complications were lack of passive fitting between the restoration and the abutment; and destructive occlusal contacts. The destructive occlusal contacts may develop during the time of wear of the prostheses material or by changes in the opposite restoration. Overloading or bending moments exerted on the prostheses via these occlusal contacts resulted in loosening or fracture in one of the implant-prostheses components. The benefit of acrylic resin or composite material in reducing stresses in the light of the possible long-term, harmful occlusal changes that might develop as a result of their wear should be considered.


Correlates of the Michigan Alcoholism Screening Test (MAST) among DWI offenders.
Examined the relationship between scores on the Michigan Alcoholism Screening Test (MAST), Psychological Screening Inventory (PSI) and Locus of Control Scale (LOC) in a selected sample of 156 apprehended drunken drivers. About 90% were identified as problem drinkers on the MAST, and 15% were screened by the PSI as requiring more intensive psychological assessment. Scores on the MAST were independent of LOC, but positively related to results on PSI scales that assess antisocial acting-out and neurotic anxiety and also to general maladjustment (mean scale elevation). A weighted composite of three PSI scales was related significantly to MAST scores, but explained only a modest proportion of variance in them. The MAST and PSI can be used to supplement one another in screening DWI offenders.


Construct validity of a proposed new diagnostic entity: Acute Suicidal Affective Disturbance (ASAD).
The current study presents initial support for the construct validity of Acute Suicidal Affective Disturbance (ASAD), a clinical entity consisting of acute suicide risk and several related features. Participants (N=195) were university students who were recruited for a history of suicide attempt(s), history of suicidal ideation, or no history of suicide attempts or suicidal ideation. Participants completed study measures online. Factor analytic results indicated a one factor solution for a lifetime measure of ASAD symptoms. The measure demonstrated strong convergent and divergent validity with common correlates of suicide-related outcomes and incremental predictive validity, as lifetime occurrence of ASAD symptoms predicted number of past suicide attempts above and beyond a host of suicide risk factors. Lifetime ASAD symptoms differed between those with multiple suicide attempts, those with a single attempt, and participants without a history of attempts, as well as between participants with a history of both suicidal ideation and attempts and those with a history of suicidal ideation but not suicide attempts. The cross-sectional research design limits the ability to infer causation between ASAD symptoms and suicidal behavior. Only past ASAD symptoms (not current symptoms) were measured. ASAD appears to be a unified clinical entity that characterizes acute suicide risk which may assist clinicians in determining a client's potential for death by suicide.


Determination of the mitotic index by microinjection of fluorescently labelled tubulin.
The microneedle injection technique is one of the most established procedures for the introduction of proteins into living cells. To analyse injected proteins which are important in cell cycle progression it is often necessary to determine the mitotic index. Measuring the mitotic index after microinjection is complicated because only a limited number of cells of the whole cell population is microinjected. Therefore, we attempted to establish a new method to determine the mitotic index using microinjection of fluorescently labelled alpha/beta-tubulin into mammalian cells which allows to monitor the injected cells simultaneously with the determination of the mitotic index. We demonstrated that fluorescently labelled tubulin incorporates efficiently into the mitotic spindle apparatus. Fluorescence remains stable for several hours which is sufficient to observe the progression of cells through the M-phase of the cell cycle. The determination of the mitotic index with the method presented here gave similar results to those determined using other methods. With this method also different stages of mitosis can be visualized by analysing various steps of spindle formation. Thus, this rapid method allows the monitoring of the injected cells after microneedle injection and simultaneously the determination of the mitotic index.


Pattern recognition receptors in grass carp Ctenopharyngodon idella: I. Organization and expression analysis of TLRs and RLRs.
Pattern recognition receptors (PRRs) play indispensable roles in the immune responses against invading pathogens. In the present study, we systematically identified and characterized Toll-like receptors (TLRs), retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) as well as their adaptors in grass carp (Ctenopharyngodon idella). A comprehensive analysis of BLAST and other bioinformatics methods showed that C. idella TLR family consist of 21 members and their adaptors contain four members. Phylogenetic analyses confirmed the existence of six TLR subfamilies (TLR1, 3, 4, 5, 7 and 11 subfamily) in C. idella and revealed their homologous relationships with other species. Most C. idella TLRs possess three typical structural features of TLR protein family: LRR, TM and TIR domains. Meanwhile, RLR family consist of three conserved members (RIG-I, MDA5 and LGP2) as well as two adaptors (IPS-1 and STING) in C. idella. mRNA expression analyses of TLRs, RLRs and their adaptors indicated that most members are sustainably expressed in multiple tissues before and after grass carp reovirus (GCRV) or Aeromonas hydrophila infection, while TLR9, TLR20a/b, TLR25, TIRAP, SARM1 and STING are transiently expressed in specific tissues. TLRs are transmembrane receptors with few introns, while RLRs are cytoplasmic receptors with plenty of introns. TLRs and RLRs interact with adaptors to perform their functions via various signaling pathways. In conclusion, this study systematically explores the characteristics of TLRs and RLRs in C. idella and provides evidence for the response patterns after viral and/or bacterial infection in vivo. These results contribute to studying the regulation mechanisms of TLR and RLR signaling pathways, and deeply understanding fish immune responses against pathogen infection.


Optimizing dendritic cell-based approaches for cancer immunotherapy.
Dendritic cells (DC) are professional antigen-presenting cells uniquely suited for cancer immunotherapy. They induce primary immune responses, potentiate the effector functions of previously primed T-lymphocytes, and orchestrate communication between innate and adaptive immunity. The remarkable diversity of cytokine activation regimens, DC maturation states, and antigen-loading strategies employed in current DC-based vaccine design reflect an evolving, but incomplete, understanding of optimal DC immunobiology. In the clinical realm, existing DC-based cancer immunotherapy efforts have yielded encouraging but inconsistent results. Despite recent U.S. Federal and Drug Administration (FDA) approval of DC-based sipuleucel-T for metastatic castration-resistant prostate cancer, clinically effective DC immunotherapy as monotherapy for a majority of tumors remains a distant goal. Recent work has identified strategies that may allow for more potent "next-generation" DC vaccines. Additionally, multimodality approaches incorporating DC-based immunotherapy may improve clinical outcomes.


Blowhold cecostomy for cecal decompression.
Blowhole cecostomy is a method for achieving decompression of the distended cecum. Emphasis is placed on a McBurney-type incision, a watertight suture line joining bowel to muscle, and avoidance of opening the cecum until the suturing is done.


HIV-GB virus C co-infection: an overview.
GB virus C (GBV-C) and hepatitis G virus (HGV) are two isolates of the same virus, independently identified in humans in the 1990s by two research laboratories, and were initially considered a potential cause of liver disease. Studies failed to associate the virus with hepatitis or any known human disease. GBV-C reappeared in the scientific scene when some research groups, in an attempt to find the interference of the virus among HIV seropositive patients, reported a lower mortality rate and slower disease progression among co-infected patients. From then on, several mechanisms have been proposed to clarify this putative benefit; however, the question whether GBV-C exerts a protective effect in HIV-infected patients remains to be resolved.


Hydrogen bonds in 1,4-dioxane/ammonia binary clusters.
With synchrotron radiation, we have studied the photoionization and dissociation of 1,4-dioxane/ammonia clusters in a supersonic expansion. The observed major product ions are the 1,4-dioxane cation M(+) and protonated cluster ions M(NH(3))(n)H(+) (where M=1,4-dioxane), and the intensities of the unprotonated cluster ions M(NH(3))(n) (+) are much lower. Fully optimized geometries and energies of the neutral cluster M(NH(3))(2) and related cluster ions have been obtained using the ab initio molecular orbital method and density functional theory. The potential energy surface of the excited state of M(NH(3))(2) (+) was also calculated. With these results, the mechanisms of different photoionization-dissociation channels have been suggested. The most probable channel is electron ejection from the highest occupied molecular orbital, followed by the dissociation into M(+) and (NH(3))(2). For another main channel, after removing an electron from the second highest occupied molecular orbital, the intracluster proton transfer process takes place to form the stable unprotonated cluster ion M(NH(3))H(+)-NH(2), which usually leads to the dissociated protonated cluster ion M(NH(3))H(+) and a radical NH(2).


Topical ocular delivery to laser-induced choroidal neovascularization by dual internalizing RGD and TAT peptide-modified nanoparticles.
A nanoparticle (NP) was developed to target choroidal neovascularization (CNV) via topical ocular administration. The NPs were prepared through conjugation of internalizing arginine-glycine-aspartic acid RGD (iRGD; Ac-CCRGDKGPDC) and transactivated transcription (TAT) (RKKRRQRRRC) peptide to polymerized ethylene glycol and lactic-co-glycolic acid. The iRGD sequence can specifically bind with integrin αvβ3, while TAT facilitates penetration through the ocular barrier. 1H nuclear magnetic resonance and high-performance liquid chromatography demonstrated that up to 80% of iRGD and TAT were conjugated to poly(ethylene glycol)- poly(lactic-co-glycolic acid). The resulting particle size was 67.0±1.7 nm, and the zeta potential of the particles was -6.63±0.43 mV. The corneal permeation of iRGD and TAT NPs increased by 5.50- and 4.56-fold compared to that of bare and iRGD-modified NPs, respectively. Cellular uptake showed that the red fluorescence intensity of iRGD and TAT NPs was highest among primary NPs and iRGD- or TAT-modified NPs. CNV was fully formed 14 days after photocoagulation in Brown Norway (BN) rats as shown by optical coherence tomography and fundus fluorescein angiography analyses. Choroidal flat mounts in BN rats showed that the red fluorescence intensity of NPs followed the order of iRGD and TAT NPs > TAT-modified NPs > iRGD-modified NPs > primary NPs. iRGD and TAT dual-modified NPs thus displayed significant targeting and penetration ability both in vitro and in vivo, indicating that it is a promising drug delivery system for managing CNV via topical ocular administration.


Vitamin C and Cell Death.
Significance: Persistent oxidative stress is a common feature of cancer cells, giving a specific weapon to selectively eliminate them. Ascorbate in pharmacological concentration can contribute to the suspended formation of hydroxyl radical via the Fenton reaction; thus, it can be an important element of the oxidative stress therapy against cancer cells. Recent Advances: The main components of ascorbate-induced cell death are DNA double-strand breaks via the production of hydroxyl radical and ATP depletion due to the activation of poly (ADP-ribose) polymerase 1. Presumably, DNA damage can be the primary contributor to the anticancer activity of pharmacological ascorbate, as opposed to the rupture of bioenergetics. The caspase independency of high-dose ascorbate-induced cell death proposed the possible involvement of several types of cell death, such as ferroptosis, necroptosis, and autophagy. Critical Issues: Ascorbate can target at least two key molecular features of cancer cells as a part of the anticancer therapy: the intrinsic or acquired resistance to cell death and the dysregulated metabolism of cancer cells. It seems probable that different concentrations of ascorbate alter the nature of induced cell death. Autophagy and necroptosis may play a role at intermediate concentrations, but caspase-independent apoptosis may dominate at higher concentrations. However, ascorbate behaves as an effective inhibitor of ferroptosis that may have crucial importance in its possible clinical application. Future Directions: The elucidation of the details and the links between high-dose ascorbate-induced cancer selective cell death mechanisms may give us a tool to form and apply synergistic cancer therapies. Antioxid. Redox Signal. 34, 831-844.


Right main bronchus rupture due to blunt chest trauma.
Bronchial rupture following major blunt chest trauma should be suspected in every case of massive and persistent air leak through the intercostal drain tube. Chest radiogram offers indirect signs, while chest CT scan demonstrates specific signs highly suggestive for this extremely rare tracheobronchial injury. Bedside bronchoscopy confirms the diagnosis.


Behavior analysis and neuroscience: Complementary disciplines.
Behavior analysis and neuroscience are disciplines in their own right but are united in that both are subfields of a common overarching field-biology. What most fundamentally unites these disciplines is a shared commitment to selectionism, the Darwinian mode of explanation. In selectionism, the order and complexity observed in nature are seen as the cumulative products of selection processes acting over time on a population of variants-favoring some and disfavoring others-with the affected variants contributing to the population on which future selections operate. In the case of behavior analysis, the central selection process is selection by reinforcement; in neuroscience it is natural selection. The two selection processes are inter-related in that selection by reinforcement is itself the product of natural selection. The present paper illustrates the complementary nature of behavior analysis and neuroscience through considering their joint contributions to three central problem areas: reinforcement-including conditioned reinforcement, stimulus control-including equivalence classes, and memory-including reminding and remembering.


Motives for exercise participation as predictors of exercise dependence among endurance athletes.
To investigate whether motives for exercise participation predicted exercise dependence (ED) among endurance athletes. The rationale for the study centred upon a test of the affect regulation model utilising constructs that form part of the Self-Determination Theory as predictors of ED. It was hypothesised that non self-determined motivation, specifically external regulation, would be predictive of ED. correlational design, with a time gap between predictor and dependent variables. competitive sports environment. 188 competitive endurance athletes were recruited from amateur sports clubs. none. the Behavioural Regulation in Exercise Questionnaire was administered before a training session to measure the predictor variables (motives for exercise participation), and the Running Addiction Scale was administered before a similar training session, one week later, to measure the dependent variable (ED). Multiple regression analysis revealed that the strongest predictor variable of ED was introjected regulation (beta=0.29, p<0.001), followed by identified regulation (beta=0.19, p<0.05). External regulation and intrinsic motivation were weak and non-significant predictors. The total variance in ED explained by the exercise participation motives was 15% (R(2)= 0.15). ED was predicted by motives that did not support the tenets of the affect regulation model. Results are discussed in light of the potential influence of exercise participation motives on ED and their implications for intervention strategies and diagnosis of the ED syndrome.


Peptide-functionalized poly(ethylene glycol) star polymers: DNA delivery vehicles with multivalent molecular architecture.
Exploring the development of nonviral nucleic acid delivery vectors with progressive, specific, and novel designs in molecular architecture is a fundamental way to investigate how aspects of chemical and physical structure impact the transfection process. In this study, macromolecules comprised of a four-arm star poly(ethylene glycol) and termini modified with one of five different heparin binding peptides have been investigated for their ability to bind, compact, and deliver DNA to mammalian cells in vitro. These new delivery vectors combine a PEG-derived stabilizing moiety with peptides that exhibit unique cell-surface binding ability in a molecular architecture that permits multivalent presentation of the cationic peptides. Five peptide sequences of varying heparin binding affinity were studied; each was found to sufficiently bind heparin for biological application. Additionally, the macromolecules were able to bind and compact DNA into particles of proper size for endocytosis. In biological studies, the PEG-star peptides displayed a range of toxicity and transfection efficiency dependent on the peptide identity. The vectors equipped with peptides of highest heparin binding affinity were found to bind DNA tightly, increase levels of cellular internalization, and display the most promising transfection qualities. Our results suggest heparin binding peptides with specific sequences hold more potential than nonspecific cationic polymers to optimize transfection efficiency while maintaining cell viability. Furthermore, the built-in multivalency of these macromolecules may allow simultaneous binding of both DNA at the core of the polyplex and heparan sulfate on the surface of the cell. This scheme may facilitate a bridging transport mechanism, tethering DNA to the surface of the cell and subsequently ushering therapeutic nucleic acids into the cell. This multivalent star shape is therefore a promising architectural feature that may be exploited in the design of future polycationic gene delivery vectors.


Exploring plant diversity through soil DNA in Thai national parks for influencing land reform and agriculture planning.
The severe deforestation, as indicated in national forest data, is a recurring problem in many areas of Northern Thailand, including Doi Suthep-Pui National Park. Agricultural expansion in these areas, is one of the major drivers of deforestation, having adverse consequences on local plant biodiversity. Conserving biodiversity is mainly dependent on the biological monitoring of species distribution and population sizes. However, the existing conventional approaches for monitoring biodiversity are rather limited. Here, we explored soil DNA at four forest types in Doi Suthep-Pui National Park in Northern Thailand. Three soil samples, composed of different soil cores mixed together, per sampling location were collected. Soil biodiversity was investigated through eDNA metabarcoding analysis using primers targeting the P6 loop of the plastid DNA trnL (UAA) intron. The distribution of taxa for each sample was found to be similar between replicates. A strong congruence between the conventional morphology- and eDNA-based data of plant diversity in the studied areas was observed. All species recorded by conventional survey with DNA data deposited in the GenBank were detected through the eDNA analysis. Moreover, traces of crops, such as lettuce, maize, wheat and soybean, which were not expected and were not visually detected in the forest area, were identified. It is noteworthy that neighboring land and areas in the studied National Park were once used for crop cultivation, and even to date there is still agricultural land within a 5-10 km radius from the forest sites where the soil samples were collected. The presence of cultivated area near the forest may suggest that we are now facing agricultural intensification leading to deforestation. Land reform for agriculture usage necessitates coordinated planning in order to preserve the forest area. In that context, the eDNA-based data would be useful for influencing policies and management towards this goal.


Experimentally Switching from Factory Made to Self-Made Cigarettes: A Preliminary Study of Perceptions, Toxicant Exposure and Smoking Behavior.
There is currently the potential for a great deal of transition and product switching among cigarette smokers. Studies on the transition when cigarette smokers switch from one type of nicotine delivery product to another are needed to understand subsequent toxicant exposure. A preliminary study was performed to determine the feasibility of experimentally replicating the transition from factory made (FM) to personal machine made (PMM) cigarette smoking. The adaptability and perceptions of the consumer and the consequent exposure to cigarette-delivered toxins were assessed. Six adults (4 men) were recruited for four laboratory visits (V1-V4) on study days 1, 5, 10 and 15, respectively. All of the participants agreed to switch from exclusive FM smoking to exclusive PMM cigarette smoking for the duration of the study. Compliance was very high among these participants. Participants progressively accepted the PMM cigarettes and became efficient producers of PMMs as evidenced in the reduced time to make 5 PMMs in the laboratory. Participants reported a preference for FM at visit 2 (V2), but had stated no preference by the fourth visit. Compared to the FMs, the PMMs at V3 (p<0.05) and V4 (p<0.10) had lower CO boost (7.3 vs. 4.1 ppm; p<0.05). Over all conditions, nicotine plasma levels averaged 18.0±2.4 ng/ml before smoking (for both FM and PMM) and 34.0±5.3 ng/mL after smoking; there were no significant differences in the plasma nicotine boost (average 17.7 and 15.4ng/ml after FM and PMM smoking, respectively). Although there were differences between individual subjects' filter butt levels of deposited solanesol the within-subject levels were remarkably similar. Puff topography measures did not vary across visits or cigarette type. Although interpretation of study results must be conservative because of the small sample size, this study demonstrates that experimentally-induced transition from FM to PMM smoking is feasible for laboratory study and the subsequent toxicant exposure is comparable for FM and PMM cigarettes.


CYP2D6-guided opioid therapy improves pain control in CYP2D6 intermediate and poor metabolizers: a pragmatic clinical trial.
CYP2D6 bioactivates codeine and tramadol, with intermediate and poor metabolizers (IMs and PMs) expected to have impaired analgesia. This pragmatic proof-of-concept trial tested the effects of CYP2D6-guided opioid prescribing on pain control. Participants with chronic pain (94% on an opioid) from seven clinics were enrolled into CYP2D6-guided (n = 235) or usual care (n = 135) arms using a cluster design. CYP2D6 phenotypes were assigned based on genotype and CYP2D6 inhibitor use, with recommendations for opioid prescribing made in the CYP2D6-guided arm. Pain was assessed at baseline and 3 months using PROMIS® measures. On stepwise multiple linear regression, the primary outcome of composite pain intensity (composite of current pain and worst and average pain in the past week) among IM/PMs initially prescribed tramadol/codeine (n = 45) had greater improvement in the CYP2D6-guided versus usual care arm (-1.01 ± 1.59 vs. -0.40 ± 1.20; adj P = 0.016); 24% of CYP2D6-guided versus 0% of usual care participants reported ≥30% (clinically meaningful) reduction in the composite outcome. In contrast, among normal metabolizers prescribed tramadol or codeine at baseline, there was no difference in the change in composite pain intensity at 3 months between CYP2D6-guided (-0.61 ± 1.39) and usual care (-0.54 ± 1.69) groups (adj P = 0.540). These data support the potential benefits of CYP2D6-guided pain management.


Subcutaneous transplantation of human embryonic stem cells-derived pituitary organoids.
The pituitary gland, regulating various hormones, is central in the endocrine system. As spontaneous recovery from hypopituitarism is rare, and exogenous-hormone substitution is clumsy, pituitary replacement via regenerative medicine, using pluripotent stem cells, is desirable. We have developed a differentiation method that in mice yields pituitary organoids (POs) derived from human embryonic stem cells (hESC). Efficacy of these POs, transplanted subcutaneously into hypopituitary mice, in reversing hypopituitarism was studied. hESC-derived POs were transplanted into inguinal subcutaneous white adipose tissue (ISWAT) and beneath dorsal skin, a relatively avascular region (AR), of hypophysectomized severe combined immunodeficient (SCID) mice. Pituitary function was evaluated thereafter for ¾ 6mo, assaying basal plasma ACTH and ACTH response to corticotropin-releasing hormone (CRH) stimulation. Histopathologic examination of organoids 150d after transplantation assessed engraftment. Some mice received an inhibitor of vascular endothelial growth factor (VEGF) to permit assessment of how angiogenesis contributed to subcutaneous engraftment. During follow-up, both basal and CRH-stimulated plasma ACTH levels were significantly higher in the ISWAT group (p < 0.001 - 0.05 and 0.001 - 0.005, respectively) than in a sham-operated group. ACTH secretion also was higher in the ISWAT group than in the AR group. Histopathologic study found ACTH-producing human pituitary-cell clusters in both groups of allografts, which had acquired a microvasculature. POs qPCR showed expression of angiogenetic factors. Plasma ACTH levels decreased with VEGF-inhibitor administration. Subcutaneous transplantation of hESC-derived POs into hypopituitary SCID mice efficaciously renders recipients ACTH-sufficient.


Let-7 and MicroRNA-148 Regulate Parathyroid Hormone Levels in Secondary Hyperparathyroidism.
Secondary hyperparathyroidism commonly complicates CKD and associates with morbidity and mortality. We profiled microRNA (miRNA) in parathyroid glands from experimental hyperparathyroidism models and patients receiving dialysis and studied the function of specific miRNAs. miRNA deep-sequencing showed that human and rodent parathyroids share similar profiles. Parathyroids from uremic and normal rats segregated on the basis of their miRNA expression profiles, and a similar finding was observed in humans. We identified parathyroid miRNAs that were dysregulated in experimental hyperparathyroidism, including miR-29, miR-21, miR-148, miR-30, and miR-141 (upregulated); and miR-10, miR-125, and miR-25 (downregulated). Inhibition of the abundant let-7 family increased parathyroid hormone (PTH) secretion in normal and uremic rats, as well as in mouse parathyroid organ cultures. Conversely, inhibition of the upregulated miR-148 family prevented the increase in serum PTH level in uremic rats and decreased levels of secreted PTH in parathyroid cultures. The evolutionary conservation of abundant miRNAs in normal parathyroid glands and the regulation of these miRNAs in secondary hyperparathyroidism indicates their importance for parathyroid function and the development of hyperparathyroidism. Specifically, let-7 and miR-148 antagonism modified PTH secretion in vivo and in vitro, implying roles for these specific miRNAs. These findings may be utilized for therapeutic interventions aimed at altering PTH expression in diseases such as osteoporosis and secondary hyperparathyroidism.


[Establishment and characterization of a cell line (H-1) from squamous cell carcinoma of human gingiva--effect of temperature].
We established a cell line (H-1) from human oral cancer. It was derived from a biopsy specimen of squamous cell carcinoma in the lower gingiva of a japanese man. The cell line has now been growing in vitro for more than 86 passages. The cells had an epithelial morphology and grew as adherent monolayers. PAS stain was positive and activity of ALP also was positive. They had a doubling time of 37 hours and colony formation of 32%. Chromosomal numbers ranged from 49 to 75 and the modal number was 61. The cell line was tumorigenic in nude mice, its histologic examination showed identification with that from the original tumor. The cells were sensitive to hyperthermia.


Effects of speed of processing training and transcranial direct current stimulation on global sleep quality and speed of processing in older adults with and without HIV: A pilot study.
Some older adults with human immunodeficiency virus (HIV) experience poor sleep which can worsen cognition. Transcranial direct current stimulation (tDCS) and cognitive training have improved sleep and cognition in studies of older adults; yet, their combined influence is unknown in adults with HIV. Older adults with HIV (n = 33) and without HIV (n = 33) were randomized to receive 10 one-hour sessions of speed of processing (SOP) training with tDCS or sham tDCS over approximately 5 weeks. tDCS with SOP training did not improve sleep. Omitting correction of multiple comparisons for this exploratory pilot study, main effects for HIV (F[1, 59] = 5.26, p = .03, ηp2 = .082) and tDCS (F[1, 59] = 5.16, p = .03, ηp2 = .080) on the Digit Copy Test were detected. A HIV × tDCS interaction was detected on the Letter Comparison Test (F[1, 59] = 5.50, p = .02, ηp2 = .085). Useful Field of View scores improved across all four groups (F[1, 59] = 64.76, p < .001, ηp2 = .523). No significant effects for HIV (F[1, 59] = 1.82, p = .18) and tDCS (F[1, 59] = .01, p = .94) were detected on the Useful Field of View test. While the current study did not show effects of combined tDCS and SOP training on sleep quality, future studies are needed to examine the effects of such interventions on sleep-related cognitive functions among cognitively impaired adults with HIV.


The Impact of Playing a Musical Instrument on Obstructive Sleep Apnea: A Systematic Review.
To conduct a systematic review of the medical literature evaluating the effects of playing a musical instrument on obstructive sleep apnea (OSA). Scopus, CINAHL, PubMed, and OVID. Searches were performed through October 22, 2019. The Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) statement was followed. After thorough investigation amidst specific exclusion criteria, four studies were included in the systematic review: one randomized controlled trial and three cross-sectional studies. Polysomnography was used to assess apnea-hypopnea index (AHI) related to obstructive sleep apnea (OSA) in the randomized controlled trial, while the Berlin questionnaire was used to assess high- and low risk OSA in the cross-sectional studies. Various instrument types were mentioned in these studies, largely focusing on wind instruments and subgroups within the wind instrument family. Current literature, although sparse, suggests that playing certain types of wind instruments are associated with either improving patients' AHI or reducing the risk of developing OSA. Future studies with larger sample sizes utilizing validated diagnostic tools to measure the presence and severity of OSA are necessary to determine the true impact of such interventions. Although more research is needed, sustainable and minimally invasive interventions such as playing a specific type of musical instrument could serve as an accessible, inexpensive, and effective adjunctive treatment for OSA.


Does fiscal decentralization improve health outcomes? Evidence from infant mortality in Italy.
Despite financial and decision-making responsibilities having been increasingly devolved to lower levels of government worldwide, the potential impact of these reforms remains largely controversial. This paper investigates the hypothesis that a shift towards a higher degree of fiscal autonomy of sub-national governments could improve health outcomes, as measured by infant mortality rates. Italy is used as a case study since responsibilities for healthcare have been decentralized to regions, though the central government still retains a key role in ensuring all citizens uniform access to health services throughout the country. A linear fixed-effects regression model with robust standard errors is employed for a panel of 20 regions over the period 1996-2012 (340 observations in the full sample). Decentralization is proxied by two different indicators, capturing the degree of decision-making autonomy in the allocation of tax revenues and the extent to which regions rely on fiscal transfers from the central government. The results show that a higher proportion of tax revenues raised and/or controlled locally as well as a lower transfer dependency from the central government are consistently associated with lower infant mortality rates, ceteris paribus. The marginal benefit from fiscal decentralization, however, is not constant but depends on the level of regional wealth, favouring poorest regions. In terms of policy implications, this study outlines how the effectiveness of decentralization in improving health outcomes is contingent on the characteristics of the context in which the process takes place.


Interventional management of gastrointestinal fistulas.
Gastrointestinal (GI) fistulas are frequently very serious complications that are associated with high morbidity and mortality. GI fistulas can cause a wide array of pathophysiological effects by allowing abnormal diversion of the GI contents, including digestive fluid, water, electrolytes, and nutrients, from either one intestine to another or from the intestine to the skin. As an alternative to surgery, recent technical advances in interventional radiology and percutaneous techniques have been shown as advantageous to lower the morbidity and mortality rate, and allow for superior accessibility to the fistulous tracts via the use of fistulography. In addition, new interventional management techniques continue to emerge. We describe the clinical and imaging features of GI fistulas and outline the interventional management of GI fistulas.


Carotid sinus nerve is involved in cardiorespiratory responses to intracarotid injection of capsaicin in the rat.
The carotid sinus nerve (CSN), important in cardiorespiratory modulation, mainly contains C fibers (CSCFs). Previous studies have demonstrated that selective stimulation of bronchopulmonary C fibers (PCFs) via right atrial injection of capsaicin (Cap; approximately 0.25 microg) results in an apnea (approximately 3 s) associated with hypotension and bradycardia. The present study was undertaken to determine the effects of activating CSCFs on cardiorespiratory activities. Intracarotid injection of Cap was performed before and after bilateral transection of the CSN in anesthetized and spontaneously breathing rats. Our results showed that 1) low doses of Cap (up to 2 ng) produced an increase in minute ventilation by elevating both tidal volume and respiratory frequency with the threshold dosage at 1.0 ng (P < 0.05); 2) high doses (4-64 ng) generated an apnea (prolongation of expiratory duration by approximately 8-fold) and hypertension (P < 0.05); 3) bilateral transection of the CSN reduced excitatory and inhibitory respiratory responses by 30 and 81%, respectively, and increased the hypertension by 88% (P < 0.05); and 4) the same doses of Cap delivered into the right atrium to stimulate PCFs failed to evoke detectable cardiorespiratory responses. Our results suggest that compared with PCFs, CSCFs are more sensitive to Cap stimulation and that activation of these fibers significantly modulates cardiorespiratory activity in anesthetized rats.


Psychosocial Determinants of Tobacco Use among School Going Adolescents in Delhi, India.
Background. Tobacco use is one of the major preventable causes of premature death and disease in the world. Many psychosocial factors were found to influence tobacco use. Therefore the present study was designed to determine the role of psychosocial factors associated with tobacco use among school going adolescents in Delhi, India. Methods. Cross-sectional study was conducted from February 2013 to September 2013 in four government schools in South district of Delhi, India. The questionnaire contains questions adapted from GYTS (Global Youth Tobacco Survey) to find the prevalence and pattern of tobacco use among adolescents. Data were analyzed using SPSS version 21. Results. The prevalence of ever and current tobacco use was found in 16.4% and 13.1%. Current smoking and current tobacco chewing were found in 10.2% and 9.4% students, respectively. The risk of current tobacco use was found to be higher among males (P value = 0.000) and in those who got higher pocket money (P value = 0.000). Psychosocial factors like lower general self-efficacy and maladjustments with peers, teachers, and schools were also found to be significant predictors of current tobacco use. Conclusion. The study has revealed higher prevalence of ever and current tobacco use among adolescent students in Delhi, India.


Epidemiology of rheumatoid arthritis in Finland.
To review the work pertaining to rheumatoid arthritis (RA) morbidity in Finland and to compare the data with that available from other countries. Extensive investigations in Finland of the epidemiology of RA, based on nationwide registers designed primarily for administrative purposes and on extensive population studies, frequently in combination. According to several surveys with somewhat different study designs, the prevalence of clinically significant RA is about 0.8% of the adult Finnish population. Five national health interviews from a 30-year period have revealed figures about 50% higher, but with no clear change in prevalence. The incidence of clinically significant RA is about 40 per 100,000 of the adult population, which is in accordance with the prevalence figures. The mean age at diagnosis increased by 7.6 years from 1975 to 1990. Between 1978 and 1980, 5.8% of the severe disability in the adult Finnish population was attributable to RA. Some evidence suggests that severe disability resulting from RA diminished during the 1980s, possibly because of joint replacement surgery. Monitoring sickness insurance statistics is a useful means of following the epidemiology of RA.


[Evaluation of cord functions in patients with cervical radiculomyelopathy using cortical and spinal somatosensory evoked potentials].
It is desirable for neurosurgeons to be able to evaluate cord functions. To assess electrophysiologically functional status of spinal cord, cortical and spinal somatosensory evoked potentials (SEP) were studied in 33 patients with cervical radiculomyelopathy; 11 cases of cervical spondylosis, 11 cases of OPLL, 3 cases of narrow canal, 2 cases of narrow canal with OPLL, 3 cases of intradural extramedullary tumor, 2 cases of intramedullary tumor and one case of traumatic cervical cord injury. Before the operation cortical SEP was recorded from post-Rolandic area by the median nerve stimulation. Spinal SEP was recorded from the dorsal surface of the cord after partial and complete laminectomy, or before and after tumor removal. Cortical SEP showed abnormalities in 45% of the cases, whose abnormalities were disappearance of the early components such as P1N1 wave, P1 to P2 complex or P1 to P3 complex and delay of the peak latency. Spinal SEP showed abnormalities in 42% of the cases, and the abnormal patterns were divided into 3 groups as follows: Type 1: Slightly abnormal, P1 and N1 waves are well recorded, whereas the P2 wave is absent or markedly suppressed in the amplitude. Type 2: Moderately abnormal, the P2 wave is flat and in addition the N1 wave is diminished in the amplitude. Type 3: Severely abnormal, only the P1 wave is recorded and the N1 to P2 complex is absent. Type 1 was obtained from 3 patients, type 2 from 9 patients, and type 3 from 2 patients. Cortical and spinal SEP were closely related to deep sensations, and cortical SEP was relatively correlated with superficial sensations. There were no differences of spinal SEP in most of the cases between partial and complete laminectomy. In intradural extramedullary tumor, however, good spinal SEP was recorded in collapsed cord after the tumor removal. Clinical symptoms were improved postoperatively in 80% of patients with both normal cortical and spinal SEP. It is concluded that cortical and spinal SEP are useful for prognostic value in patients with cervical radiculomyelopathy.


Determination of uric acid in serum by SERS system based on VO-MnCo2O4/Ag nanozyme.
The level of uric acid is crucial to human health. Octahedral oxygen vacancy MnCo2O4/Ag (VO-MnCo2O4/Ag) nanozyme was successfully prepared by simple hydrothermal, calcination and self-reduction methods. VO-MnCo2O4/Ag nanozyme is rich in Mn2+/Mn3+ and CO2+/CO3+ redox electron pairs, large specific surface area and oxygen vacancies. VO-MnCo2O4/Ag nanozyme showed high uricase-like activity and peroxidase-like activity. At the same time, the SERS signal of the detected molecule could be significantly enhanced after the catalytic reaction of the VO-MnCo2O4/Ag nanozyme. The Km values of VO-MnCo2O4/Ag nanozyme for H2O2 and TMB were 0.04 mM and 0.027 mM respectively. Based on the uric acid oxidase-like and peroxidase-like activities of VO-MnCo2O4/Ag, we developed a label-free, sensitive, and reliable SERS uric acid detection system. The detection linear range of uric acid is 0.01 μM-1000 μM and the detection of limit is 7.8 × 10-9 M. The results show that the sensing system has good accuracy, sensitivity, selectivity, and stability. It can be applied to the determination of samples under different conditions. This study provides profound insights into the design of enzyme-like activity regulation and SERS properties regulation of nanozymes, provides guidance for the study of reaction kinetics and catalytic mechanism of nanozymes, and has broad application prospects in the field of nanozymes and SERS sensing analysis.


Reactions of sulfenic acid with 2-mercaptoethanol: a mechanism for the inhibition of gastric (H+-K+)-adenosine triphosphate by omeprazole.
The reactions of omeprazole, a potent proton pump inhibitor (PPI) were investigated in the presence of 2-mercapotoethanol. Reactions were monitored in solutions buffered to pH values ranging 2.0-8.0 using differential pulse polarography (DPP) at the static mercury drop electrode (SMDE). The fast, sensitive and selective electrochemical technique facilitated successive recordings of voltammograms (peak current (nA) vs. peak potential (volts vs. Ag/AgCl)) for all analytes in situe, including the 2-mercaptoethanol. In acidic solutions and in the presence of 2-mercaptoethanol, omeprazole undergoes degradation into three compounds, the first is a cyclic sulfenamide (D+), previously believed to be the active inhibitor of the H+, K+-ATPase, the second is the omeprazole dimer, and the third is the disulfide believed to be the product of reaction between 2-mercaptoethanol and D+. The cyclic sulfenamide (D+) solution was found to be stable in solutions containing 2-mercaptoethanol having pH values: 2.0, 4.0, and 6.0. This finding proved conclusively that the cyclic sulfenamide is not reactive toward the 2-mercaptoethanol. In contrast to previous reports, the conversion of the sulfenic acid intermediate into D+ was found to be irreversible. Due to this irreversibility, D+ and sulfenic acid were not rapidly interconvertable. The present work suggests that the active inhibitor is the sulfenic acid.


Binding of Actinomyces viscosus to collagen: association with the type 1 fimbrial adhesin.
Treatment of hydroxyapatite (HA) with human type I or type III collagen strongly promoted adhesion of Actinomyces viscosus LY7 cells. Treatment with human type V collagen was somewhat less effective while treatment with human type IV or rat type I collagen was significantly less effective. Electron microscopic observations revealed that A. viscosus cells also attached to fibrils prepared from human type I collagen. The alpha 1 (1) polypeptide chain derived from type I collagen was ineffective in promoting binding and the alpha 2 (1) polypeptide chain exhibited moderate activity. Heat- or urea-denatured type I collagens were also ineffective in promoting binding. Mutants of A. viscosus that possess type 1 fimbriae, but not type 2 fimbriae or no fimbriae, also bound to collagen-treated HA; this suggests that the adhesin responsible was associated with type 1 fimbriae. Strains of Actinomyces israelii and Actinomyces odontolyticus also exhibited strong binding to collagen-treated HA, while Actinomyces naeslundii ATCC 12104 did not. The avidity of Actinomyces species for collagen would seem to be at least partially responsible for the high proportions of these organisms found on cemental and root tooth surfaces.


Abruptly Autofocusing Vortex Beams for Rapid Controllable Femtosecond Two-Photon Polymerization.
Micro-fabrication based on structured-beam-assisted Two-Photon Polymerization (2 PP) provides a rapid and flexible method for the manufacture of microstructures with complex morphologies. The tunable Abruptly Autofocusing Vortex (AAFV) beams were designed theoretically and generated experimentally based on a single-phase-only Spatial Light Modulator (SLM). Their specific spatial intensity distributions were further utilized to assist the fabrication of a bowl-shaped Three-Dimensional (3D) micro-trap array via 2 PP with a one-step exposure technique. Finally, the fabricated microstructures act as a novel tool for the trapping and spatial positioning of micro-particles with different diameters, which shows potential applications in fiber optics and cell study.


Perceptions by adult children of elderly parents' needs.
The purpose of this study was to examine the accuracy of adult children's perceptions of their elderly parents' concerns. Factors associated with moderate involvement between the elderly parent and adult child were predicted to lead to the greatest accuracy. 48 pairs of elderly parents and adult children filled out questionnaires regarding the parents' concerns about seven areas of functioning. Analysis showed that concerns about health and emotional support were rated as most important by elderly parents. Adult children accurately perceived these concerns to be most important to their parents. For health concerns, opposite-sex pairs of parents and children showed greatest agreement. For emotional support concerns, however, both sons and daughters were more accurate predicting their fathers' concerns than their mothers'. Middle-born children tended to be more accurate in their predictions of parents' concerns than oldest or youngest children. Living arrangements also influenced accuracy. In predicting the concerns of elderly parents, adult children were most accurate for parents living in retirement communities, moderately accurate for parents living independently, and least accurate for parents living in the adult child's home. Further, children sharing a household with an adult parent tended to overestimate their parents' concerns on some issues. The results were consistent with the moderate involvement hypothesis. The implications of these results and suggestions for research are discussed. A version of this paper was presented at the Ninety-ninth Annual Convention of the American Psychological Association in San Francisco, August 18, 1991.


Immunoproteomic characterisation of Mycoplasma mycoides subspecies capri by mass spectrometry analysis of two-dimensional electrophoresis spots and western blot.
Mycoplasma mycoides subspecies capri is one of the causative agents of contagious agalactia in goats. The disease is characterised by mastitis, pneumonia, arthritis, keratitis and in acute cases septicaemia. No vaccine is currently available that has been demonstrated to prevent disease. This study used two-dimensional electrophoresis to separate proteins from whole-cell preparations and tandem mass spectrometry to identify them. In total, 145 spots were successfully identified corresponding to 74 protein identities. Twenty of these proteins were found to be immunogenic by western blot analysis using a pooled serum sample from experimentally infected goats. Six proteins were found to have a less than 95% amino acid similarity to a closely related Mycoplasma species showing that they warrant further evaluation in development of diagnostic tests. These proteins were a dihydrolipoamide acetyltransferase component of the pyruvate dehydrogenase complex, phosphoglycerate kinase, pyrimidine-nucleoside phosphorylase, 30S ribosomal protein S6, ribulose-phosphate 3-epimerase and D-lactate dehydrogenase.


Stimulation of central serotonin turnover by beta-adrenoceptor agonists.
The beta 2-stimulators salbutamol (0.3--30 mg/kg i.v.) and clenbuterol (0.3 and 1 mg/kg i.v.), and, to a lesser extent, the beta 1-stimulators dobutamine (30 mg/kg i.v.) and prenalterol (30 mg/kg i.v.) increased serotonin metabolism in several rat brain areas, as indicated by increased concentrations of 5-hydroxyindoleacetic acid (5-HIAA) or increased tryptophan hydroxylation in vivo. With salbutamol, increases in 5-HIAA in c. striatum and brainstem, but not in cortex, were observed after intraventricular administration of relatively low doses (3--30 micrograms). Direct application of the compound into the dorsal raphe nucleus at doses of 100 ng and 1 microgram were without effect, and only minimal 5-HIAA increases occurred after the high dose of 10 micrograms. The effects of salbutamol on the concentrations of 5-HIAA were antagonized by both propranolol and WB4101, indicating an involvement not only of beta-receptors but also of postsynaptic alpha-receptors. The evidence for an against a central site of action of beta-agonists with respect to their effect on serotonergic systems is discussed.


Suppressor analysis points to the subtle role of the LAG1 ceramide synthase gene in determining yeast longevity.
Individual yeast cells display a finite replicative capacity. LAG1 was identified as a gene that is differentially expressed during the yeast replicative life span and was shown to play a role in determining yeast longevity. This gene is not essential, but simultaneous deletion of LAG1 and its close homologue LAC1 is lethal. Lag1p and Lac1p have been found to be an essential component of ceramide synthase. In this study, multicopy suppressors of the lethality of a lag1delta lac1delta double mutant were isolated to help clarify the role of LAG1 in yeast longevity. The two multicopy suppressors YBR183w (YPC1) and YPL087w (YDC1) encode ceramidases unrelated to Lag1p and Lac1p, which were previously found to support the reverse reaction of ceramide synthesis. Multiple copies of YPC1 were much more efficient than YDC1 in rescuing cell growth. They were also much more effective in rescuing the life span of a lag1delta lac1delta double mutant, sustaining a life span approaching that obtained by the restoration of LAG1 expression. Neither deletion of LAC1 nor overexpression of YPC1 had a detectable effect on wild-type life span. However, the overexpression of LAG1 had a bimodal effect on longevity, with moderate expression resulting in increased longevity and with higher expression curtailing life span. These results suggest that subtle changes in ceramide/sphingolipid metabolism are important in determining yeast longevity. They also indicate that Lag1p plays a special role in this relationship. Homologues of Lag1p have been identified in higher eukaryotes, including human, raising the possibility that ceramide and other sphingolipid metabolites play a wider role in biological aging.


Child temperament moderates effects of parent-child mutuality on self-regulation: a relationship-based path for emotionally negative infants.
This study examined infants' negative emotionality as moderating the effect of parent-child mutually responsive orientation (MRO) on children's self-regulation (n=102). Negative emotionality was observed in anger-eliciting episodes and in interactions with parents at 7 months. MRO was coded in naturalistic interactions at 15 months. Self-regulation was measured at 25 months in effortful control battery and as self-regulated compliance to parental requests and prohibitions. Negative emotionality moderated the effects of mother-child, but not father-child, MRO. Highly negative infants were less self-regulated when they were in unresponsive relationships (low MRO), but more self-regulated when in responsive relationships (high MRO). For infants not prone to negative emotionality, there was no link between MRO and self-regulation. The "regions of significance" analysis supported the differential susceptibility model not the diathesis-stress model.


An "ecological" action-based synthesis.
We expand upon Morsella et al.'s synthesis in the direction of what Gibson (1979) called an ecological approach to perception. Morsella et al. describe consciousness as a director of voluntary action, but they understate the role of the environment in its evolution as well as in directing behavior. We elaborate these roles in the context of the concept of affordances.


[A brief textual research on the editions of Fu Ren Liang Fang (A complete book of effective prescriptions for women) before the Qing Dynasty].
Fu Ren Liang Fang (A Complete Book of Effective Prescriptions for Women), the first complete gynecologic and obstetric work in China, was written by Cheng Zi-ming in the first year of Jiaxi Reign of the Southern Song Dynasty (A.D.1237). It has many different editions in its long circulation of more than 760 years. According to The National Associated Catalog of TCM Books, it is estimated that there are as many as over 30 different editions. After choosing the edition of Qingyou Hall of the Yuan Dynasty as the original edition for comparisons with other editions, it is claimed that the edition of this Qingyou Hall was well preserved, with excellent and clear handwriting, reflecting the style and features of Chen's original work.


[Aetiology of rheumatoid arthritis].
Although the aetiology of rheumatoid arthritis(RA) is still unclear, accumulating evidences are clarifying the involvement of genetic and environmental factors in the disease, or the pathogenic mechanism of perpetuating proliferative synovitis and bone destruction. Genetic factors including HLA-DR, PADI4, death receptor 3 or DEK were reported, whereas low concordance rate of RA in monozygotic twins or other evidences strongly imply the involvement of environmental factors in RA. Each constituents of RA synovium including macrophages, stromal cells, T cells and B cells, is shown to be involved in the aetiology of RA. Recently osteoclasts are shown to be mainly involved in the bone-destruction of RA. Clarification of RA aetiology will provide alternative in RA treatment especially for refractory RA.


Structure and immune recognition of trimeric pre-fusion HIV-1 Env.
The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. Here we report the structure at 3.5 Å resolution for an HIV-1 Env trimer captured in a mature closed state by antibodies PGT122 and 35O22. This structure reveals the pre-fusion conformation of gp41, indicates rearrangements needed for fusion activation, and defines parameters of immune evasion and immune recognition. Pre-fusion gp41 encircles amino- and carboxy-terminal strands of gp120 with four helices that form a membrane-proximal collar, fastened by insertion of a fusion peptide-proximal methionine into a gp41-tryptophan clasp. Spike rearrangements required for entry involve opening the clasp and expelling the termini. N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation.


Monitoring metal levels in water and multiple biomarkers in the grouper ( Epinephelus tauvina) to assess environmental stressors on the Arabian Gulf coast of Saudi Arabia.
In this field study, the levels of heavy metals (Pb, Fe, Co, Cu, and Zn) in water and a suite of biochemical and histological biomarkers in the grouper ( Epinephelus tauvina) were assessed at four sites in the Arabian Gulf. Samples were taken from a relatively non-urban reference site, called Salwa (S1), and three effluent-dominated sites, namely Al-aziziyah in southern Dammam city (S2), the Al-Jubail coast (S3), and Manifa (S4). Toxic metals, namely Pb and Co (at all sites) and Fe (at S3), were elevated in water samples relative to the internationally permissible limits. In fish, induced levels of heat shock protein 70 (HSP70) in the liver at S3 and S4 were higher than those of the reference fish at S1. Additionally, the level of the lipid peroxidation (LPO) product (malondialdehyde (MDA)) was significantly increased in gills (at S3) and liver (at S2 and S3). There was an inhibition of catalase activities in the gills of fish from S2 to S4 and significantly higher activity levels of superoxide dismutase in the gills of fish from S4. Histopathological features such as aneurysms in gill vessels, deformed gill lamellae, increases in liver melano-macrophage centers, and hepatocellular necrosis were most abundant at sites where significant pollution problems exist (i.e. S2-S4). The results reveal that the eastern coast of Saudi Arabia, in the Arabian Gulf, is still contaminated, as indicated by elevated HSP70, LPO content and numbers of histological lesions, and that monitoring of contaminants and their effects should be continued in this region.


TLR4 and LPS hyporesponsiveness in humans.
Asthma is a complex genetic disorder that is caused by a number of unique gene-gene and gene-environment interactions. The search for asthma susceptibility genes has been complicated by the broad clinical phenotype of asthma, the polygenic inheritance pattern of this disease, and the substantial role of environmental exposures in the development and progression of asthma. Inhaled environmental agents induce several biologic responses in asthmatics; including the induction of acquired and innate immunity that leads to acute and chronic forms of airway inflammation and airway remodeling. Acquired immune responses to protein antigens, such as house dust mite allergen, often induce type 2 T lymphocyte-driven responses (Th2) which appear to be important in atopic asthma. Recent studies by our group and others demonstrate that innate immunity, initiated by inhalation of bacterial and viral pathogens, organic dusts, endotoxin or lipopolysaccharide (LPS), air pollution particulate matter, and ozone, can also cause acute and chronic forms of airflow obstruction, airway inflammation, and even airway remodeling. Emerging evidence indicates that both acquired and innate immune responses in the lung may be influenced by polymorphic genes. For instance, functional polymorphisms in the IL-4 receptor gene are thought to preferentially stimulate acquired Th2 immune responses to inhaled allergens, and we have recently shown that common co-segregating mutations in TLR4 (a transmembrane receptor for LPS) are associated with diminished airway responsiveness to inhaled LPS. These observations suggest that environmental challenges can be used to narrow the phenotype of asthma and allow scientists to investigate unique gene-environment interactions that are involved in the development of biologically specific forms of asthma.


Vital signs--the six-minute warnings.
The six classic vital signs (blood pressure, pulse, temperature, respiration, height, and weight) are reviewed on an historical basis and on their current use in dentistry. Normal and abnormal vital signs are explored in depth, with special attention to those conditions which can be readily recognized by the dental practitioner and with special attention to safe management of the patient in each instance. Measurement of blood pressure at the initial physical evaluation of the adult dental patient is approaching a standard of care status. A plea is made for expanding this practice and adding additional vital signs to usual office measurements. An efficient routine by which the dental assistant can measure and record the six vital signs in from three to six minutes is reviewed, and the frequency of vital sign measurement is explored.


Apical ballooning syndrome or Takotsubo cardiomyopathy: a new challenge in acute cardiac care.
Apical ballooning syndrome (ABS) is a unique acute cardiac syndrome characterized by symptoms and electrocardiographic changes that mimic acute myocardial infarction. It occurs in patients without evidence of significant obstructive coronary artery disease and is associated with transient extensive wall motion abnormalities of the apical and mid portions of the left ventricle. The onset of ABS is preceded by a stressful event, either emotional or physical in around 65% of cases. The underlying pathophysiology for ABS remains unclear; however, several mechanisms have been proposed including multivessel epicardial spasm, microvascular spasm, catecholamine induced myocardial stunning and myocarditis. The treatment of ABS remains entirely empirical and should be individualized according to the patient's clinical picture at the time of presentation. It should be initially managed according to the guidelines for acute coronary syndrome. Once the diagnosis of ABS is made, supportive care usually leads to spontaneous recovery. The prognosis of patients with Takotsubo cardiomyopathy is generally favourable. The left ventricular systolic dysfunction usually resolves within a few weeks. In-hospital mortality is low, less than 2%, and recurrence rate is no more than 10%. The aim of this article is to clarify, for the clinicians dealing with acute cardiac care, when they should suspect ABS and how they should confirm the diagnosis and subsequently manage it.


Extended-spectrum beta-lactamases: an actual problem of hospital microbiology (a review).
Although there is a variety of mechanisms of bacterial resistance to beta-lactam antibiotics, the most important one is production of beta-lactamases inactivating penicillins and cephalosporins. The classification of beta-lactamases is based on biochemical, enzymological (i.e. molecular structure, inhibitory property, substrate-profile, relative rate of hydrolysis) and immunological characters. Extended-spectrum beta-lactamases (ESBLs) can be derived from TEM or SHV enzymes. These enzymes have now been sequenced and it has been found that relatively few point mutations have occurred in the gene of the TEM and SHV type enzymes. These point mutations clustered in five areas of the gene. The amino acid mutations can alter the conformation, the active site and change the hydrance of beta-lactamase-cephalosporin binding capacity. So the enzyme is able to bind and hydrolyse the third generation cephalosporins. Successive mutation interacted radically increasing the binding capacity of enzymes and confer resistance to newer cephalosporins. The use of these drugs provides a strong selective pressure to develop these mutations. Sporadic nosocomial outbreaks due to strains producing an ESBL led to an epidemic problem in some hospitals resulting in a concurrent dissemination of genes, plasmids or strains. Clinical epidemiological importance and role of ESBLs and emergence of multiply resistance of bacteria of nosocomial importance are discussed in this brief.


The limitations of population health as a model for a new public health.
Population health as developed by the Canadian Institute for Advanced Research (CIAR) has influenced the shape and direction of Canadian public health policy, and has the potential to do so in the USA and elsewhere. There is reason to be concerned about this ascendence of CIAR thinking: population health is rooted within epidemiology, a militantly quantitative discipline; population health eschews analysis of societal structures as determinants of health; and population health elevates scientific understanding over health promotion action. Its lack of an explicit values base is also problematic. Policy makers should recognize these and other limitations as they consider models for a new public health.


Hyperinfection by Strongyloides stercoralis probably associated with Rituximab in a patient with mantle cell lymphoma and hyper eosinophilia.
The first report to our knowledge, of hyperinfection by Strongyloides stercoralis (HS) and hypereosinophilia, associated to immune suppression by Rituximab (the only drug received for the last one year and 10 months), in a patient with mantle-cell lymphoma (MCL), is presented. The patient has a 3-year history of MCL, and developed two accesses of HS during 2008, including meningitis, pneumonia and presence of larvae of S. stercoralis in the lungs. We had a unique chance to look at cytotoxicity of filariform larvae in the expectoration after Ivermectin treatment, showing immobilization and death of larvae, associated with eosinophils attached to the cuticle of the parasite.