A preliminary report about the detection of ventricular repolarisation in patients with non-alcoholic fatty liver disease.

OBJECTIVES: Non-alcoholic fatty liver disease (NAFLD) is a multisystem disease which can affect the cardiovascular system as well. We conducted this study to determine the cardiac effects of NAFLD such as conduction of impulse and ventricular repolarisation on electrocardiography (ECG). METHODS: In this study, we recruited patients with risk factors for NAFLD (group I; n = 23) and NAFLD patients (group II; n = 74) from Shar Hospital in Sulaimani City, Iraq. We analysed anthropometric measurements, serum fasting lipid profile, glucose levels, liver enzymes, and ECG recordings. RESULTS: ECG recordings showed significantly longer PR intervals, significantly shorter QTcB and JTc intervals, and a higher Tp-e/QTcB ratio in group II patients than in group I patients. These abnormalities were not associated with risk factors for diabetes. The TQ duration was significantly correlated with serum alanine aminotransferase (r = 0.411, p < 0.001) and aspartate aminotransferase (r = 0.272, p = 0.019) levels. CONCLUSION: In our study, the presence of significant abnormalities in ventricular repolarisation suggests that patients with newly diagnosed NAFLD have subclinical cardiac stress and a higher risk of ventricular arrhythmias.

Mechano-responsive microcapsules with uniform thin shells.

Capsules often prolong the shelf-life of active ingredients, such as many types of drugs, food additives, or cosmetic substances, because they delay oxidation of these substances or prevent their reactions with molecules contained in the surrounding. If capsules are appropriately designed, they can offer an additional benefit: they allow close control over the timing and location of the release of active ingredients. To take advantage of these features, capsules must possess shells whose thickness and composition are well-defined. However, the shell thickness of capsules often varies considerably even within a single capsule, thereby hampering good control over the release kinetics of encapsulants. These variations can be reduced, and hence the degree of control over the release kinetics increased, if shells are made thin. Unfortunately, the controlled fabrication of mechanically stable microcapsules with well-defined sub-µm thick shells is difficult. Here, we introduce a method to fabricate capsules with uniform semi-permeable shells with a thickness as low as 400 nm. This is achieved using water-oil-water double emulsions with 800 nm thick shells as templates to fabricate capsules with uniform 400 nm thin shells. These shells occupy less than 2% of the capsule volume, thereby minimizing their footprint. Despite their thin shells, these capsules are mechanically robust: they withstand pressures up to 1.3 MPa without deformation and remain intact if exposed to pressures up to 2.75 MPa. Moreover, while they are permeable towards water, they retain low molecular weight encapsulants even if dried and re-dispersed. The thin shells of the capsules open up new possibilities of their use to functionalize materials with at least one dimension that is small, such as coatings, where thick shells introduce defects, or as building blocks of new types of functional materials.

Scalable production of double emulsion drops with thin shells.

Double emulsions are often used as containers to perform high throughput screening assays and as templates for capsules. These applications require double emulsions to be mechanically stable such that they do not coalesce during processing and storage. A possibility to increase their stability is to reduce the thickness of their shells to sufficiently low values that lubrication effects hinder coalescence. However, the controlled fabrication of double emulsions with such thin shells is difficult. Here, we introduce a new microfluidic device, the aspiration device, that reduces the shell thickness of double emulsions down to 240 nm at a high throughput; thereby, the shell volume is reduced by up to 95%. The shell thickness of the resulting double emulsions depends on the pressure profile in the device and hence on the fluid flow rates in the channels and is independent of the shell thickness of the injected double emulsions. Therefore, this device enables converting double emulsions with polydisperse shell thicknesses into double emulsions with well-defined, uniform thin shells.

Prevalence of urinary incontinence and probable risk factors in a sample of kurdish women.

OBJECTIVES: The most common manifestation of pelvic floor dysfunction is urinary incontinence (UI) which affects 15-50% of adult women depending on the age and risk factors of the population studied. The aim of this study was to determine the probable risk factors associated with UI; the characteristics of women with UI; describe the types of UI, and determine its prevalence. METHODS: A cross-sectional study was conducted between February and August 2011, in the Maternity Teaching Hospital of the Erbil Governorate, Kurdistan Region, northern Iraq. It included 1,107 women who were accompanying patients admitted to the hospital. A questionnaire designed by the researchers was used for data collection. A chi-square test was used to test the significance of the association between UI and different risk factors. Binary logistic regression was used, considering UI as the dependent variable. RESULTS: The overall prevalence of UI was 51.7%. The prevalence of stress, urgency, and mixed UI was 5.4%, 13.3% and 33%, respectively. There was a significant positive association between UI and menopause, multiparity, diabetes mellitus (DM), chronic cough, constipation, and a history of gynaecological surgery, while a significant negative association was detected between UI and a history of delivery by both vaginal delivery and Caesarean section. CONCLUSION: A high prevalence of UI was detected in the studied sample, and the most probable risk factors were multiparity, menopausal status, constipation, chronic cough, and DM.

Microwave irradiation affects gene expression in plants.

The physiological impact of nonionizing radiation has long been considered negligible. However, here we use a carefully calibrated stimulation system that mimics the characteristics (isotropy and homogeneity) of electromagnetic fields present in the environment to measure changes in a molecular marker (mRNA encoding the stress-related bZIP transcription factor), and show that low amplitude, short duration, 900 MHz EMF evokes the accumulation of this mRNA. Accumulation is rapid (peaking 5-15 min after stimulation) and strong (3.5-fold), and is similar to that evoked by mechanical stimulations.

Affinity chromatography of polyhistidine tagged enzymes. New dextran-coated immobilized metal ion affinity chromatography matrices for prevention of undesired multipoint adsorptions.

New immobilized metal ion affinity chromatography (IMAC) matrices containing a high concentration of metal-chelate moieties and completely coated with inert flexible and hydrophilic dextrans are here proposed to improve the purification of polyhistidine (poly-His) tagged proteins. The purification of an interesting recombinant multimeric enzyme (a thermoresistant beta-galactosidase from Thermus sp. strain T2) has been used to check the performance of these new chromatographic media. IMAC supports with a high concentration (and surface density) of metal chelate groups promote a rapid adsorption of poly-His tagged proteins during IMAC. However, these supports also favor the promotion of undesirable multi-punctual adsorptions and problems may arise for the simple and effective purification of poly-His tagged proteins: (a) more than 30% of the natural proteins contained in crude extracts from E. coli become adsorbed, in addition to our target recombinant protein, on these IMAC supports via multipoint weak adsorptions; (b) the multimeric poly-His tagged enzyme may become adsorbed via several poly-His tags belonging to different subunits. In this way, desorption of the pure enzyme from the support may become quite difficult (e.g., it is not fully desorbed from the support even using 200 mM of imidazole). The coating of these IMAC supports with dextrans greatly reduces these undesired multi-point adsorptions: (i) less than 2% of natural proteins contained in crude extracts are now adsorbed on these novel supports; and (ii) the target multimeric enzyme may be fully desorbed from the support using 60 mM imidazole. In spite of this dramatic reduction of multi-point interactions, this dextran coating hardly affects the rate of the one-point adsorption of poly-His tagged proteins (80% of the rate of adsorption compared to uncoated supports). Therefore, this dextran coating of chromatographic matrices seems to allow the formation of strong one-point adsorptions that involve small areas of the protein and support surface. However, the dextran coating seems to have dramatic effects for the prevention of weak or strong multipoint interactions that should involve a high geometrical congruence between the enzyme and the support surface.

Molecular cloning and characterization of a tomato cDNA encoding a systemically wound-inducible bZIP DNA-binding protein.

Localized wounding of one leaf in intact tomato (Lycopersicon esculentum Mill.) plants triggers rapid systemic transcriptional responses that might be involved in defense. To better understand the mechanism(s) of intercellular signal transmission in wounded tomatoes, and to identify the array of genes systemically up-regulated by wounding, a subtractive cDNA library for wounded tomato leaves was constructed. A novel cDNA clone (designated LebZIP1) encoding a DNA-binding protein was isolated and identified. This clone appears to be encoded by a single gene, and belongs to the family of basic leucine zipper domain (bZIP) transcription factors shown to be up-regulated by cold and dark treatments. Analysis of the mRNA levels suggests that the transcript for LebZIP1 is both organ-specific and up-regulated by wounding. In wounded wild-type tomatoes, the LebZIP1 mRNA levels in distant tissue were maximally up-regulated within only 5 min following localized wounding. Exogenous abscisic acid (ABA) prevented the rapid wound-induced increase in LebZIP1 mRNA levels, while the basal levels of LebZIP1 transcripts were higher in the ABA mutants notabilis (not), sitiens (sit), and flacca (flc), and wound-induced increases were greater in the ABA-deficient mutants. Together, these results suggest that ABA acts to curtail the wound-induced synthesis of LebZIP1 mRNA.

Rapid and systemic accumulation of chloroplast mRNA-binding protein transcripts after flame stimulus in tomato.

It has been shown that tomato (Lycopersicon esculentum) plants respond to flame wounding and electrical stimulation by a rapid (15 min) and systemic up-regulation of proteinase inhibitor (pin) genes. To find other genes having a similar expression pattern, we used subtractive cDNA screening between flamed and control plants to select clones up-regulated by flame wounding. We report the characterization of one of them, a chloroplast mRNA-binding protein encoded by a single gene and expressed preferentially in the leaves. Systemic gene expression in response to flaming in the youngest terminal leaf exhibited three distinct phases: a rapid and transient increase (5-15 min) in transcript accumulation, a decline to basal levels (15-45 min), and then a second, more prolonged increase (60-90 min). In contrast, after a mechanical wound the rapid, transient increase (5 min) was followed by a rapid decline to basal levels but no later, prolonged accumulation. In the petiole, the initial flame-wound-evoked transient increase (15 min) was followed by a continuous decline for 3 h. The nature of the wound signal(s) causing such rapid changes in transcript abundance is discussed in relation to electrical signaling, which has recently been implicated in plant responses to wounding.

Rate of incidence of hepatocellular carcinoma in patients with compensated viral cirrhosis.

BACKGROUND: Cirrhosis of viral etiology due to hepatitis B virus (HBV) or hepatitis C virus (HCV) infection is a risk factor for hepatocellular carcinoma (HCC). The current study evaluated the rate of incidence of HCC in patients with compensated cirrhosis of viral etiology. METHODS: Two hundred fifty-nine cirrhotic patients (66 hepatitis B surface antigen [HBsAg] positive, 166 HCV positive, and 27 HBsAg/HCV positive) were longitudinally examined every 6 months by serum alpha-fetoprotein test and liver ultrasonography. The rates of incidence of HCC were calculated by the person-years method. The Kaplan-Meier method was used to estimate the cumulative probability of HCC development. Differences in survival time were evaluated by a log rank test. Independent predictors of HCC development were estimated by Cox proportional hazard regression analysis. RESULTS: During a mean follow-up of 64.5 months, HCC developed in 51 (19.7%) patients: in 34 of 166 HCV positive subjects (20.5%) (mean follow-up, 66.3 months), in 6 of 66 of those HBsAg positive (9.1%) (mean follow-up, 55.06 months), and in 11 of 27 of those with dual HBsAg/HCV infection (40.7%) (mean follow-up, 76.4 months). The rate of incidence of HCC per 100 person-years of follow-up was 3.7 in HCV positive subjects, 2.0 in those HBsAg positive, and 6.4 in those with dual infection. Cumulative HCC appearance rates in HBsAg positive, HCV positive, and HBsAg/HCV positive subgroups were 10%, 21%, and 23% at 5 years, 16%, 28%, and 45% at 10 years, and 16%, 40%, and 55% at 13 years, respectively. Multivariate analysis indicated that age >50 years (hazard risk [HR], 4.5; 95% confidence interval [CI] = 2.1-9.4), male gender (HR, 2.8; 95% CI = 1.1-5.3), and HBsAg/HCV coinfection (HR, 2.3; 95% CI = 1.1-4.6) were independent predictors of HCC development. CONCLUSIONS: These findings confirm that male gender and more advanced age (>50 years) are risk factors for HCC in patients with cirrhosis. Furthermore, the data indicate that subjects with dual HBsAg/HCV infection are at highest risk for HCC. Surveillance programs for early detection of HCC should focus especially on these patients.

Structure of the beta-galactosidase gene from Thermus sp. strain T2: expression in Escherichia coli and purification in a single step of an active fusion protein.

The nucleotide sequence of both the bgaA gene, coding for a thermostable beta-galactosidase of Thermus sp. strain T2, and its flanking regions was determined. The deduced amino acid sequence of the enzyme predicts a polypeptide of 645 amino acids (Mr, 73,595). Comparative analysis of the open reading frames located in the flanking regions of the bgaA gene revealed that they might encode proteins involved in the transport and hydrolysis of sugars. The observed homology between the deduced amino acid sequences of BgaA and the beta-galactosidase of Bacillus stearothermophilus allows us to classify the new enzyme within family 42 of glycosyl hydrolases. BgaA was overexpressed in its active form in Escherichia coli, but more interestingly, an active chimeric beta-galactosidase was constructed by fusing the BgaA protein to the choline-binding domain of the major pneumococcal autolysin. This chimera illustrates a novel approach for producing an active and thermostable hybrid enzyme that can be purified in a single step by affinity chromatography on DEAE-cellulose, retaining the catalytic properties of the native enzyme. The chimeric enzyme showed a specific activity of 191,000 U/mg at 70 degrees C and a Km value of 1.6 mM with o-nitrophenyl-beta-D-galactopyranoside as a substrate, and it retained 50% of its initial activity after 1 h of incubation at 70 degrees C.

Interferon treatment in hepatitis B surface antigen-positive hepatitis B e antibody-positive chronic hepatitis B: role of hepatitis B core antibody IgM titre in patient selection and treatment monitoring.

Chronic hepatitis B infection with the hepatitis B e antigen (HBeAg)-negative variant is associated with a severe clinical course and a low response rate to interferon (IFN). In an attempt to improve the chances of sustained response to interferon we designed a pilot study, using titres of IgM antibodies to hepatitis B core antigen (HBcAb IgM) to guide treatment initiation. Eighteen adults who were HBeAg-negative with biopsy-proven chronic active hepatitis (seven with cirrhosis) entered the study. They were followed-up bimonthly with routine liver function tests, and HBcAb IgM titres were also determined. Treatment (lymphoblastoid IFN 5 million units (MU) m(-2) three times weekly for 6 months) was started when the HBcAb IgM titre was increasing. Fifteen (83.3%) patients had normal alanine aminotransferase (ALT) levels and undetectable HBV DNA at the end of treatment. HBcAb IgM decreased in all responders. We observed a relapse in four patients (three with cirrhosis), in the first year after treatment, with an increase in ALT, HBV DNA and titre of HBcAb IgM. Eleven patients (61.1%) had a sustained response and eight of these 11 patients were followed-up for more than 18 months; two responders cleared hepatitis B surface antigen (HBsAg). Hence, the rate of sustained response to IFN in HBeAb-positive patients with chronic hepatitis is improved if treatment is started when HBcAb IgM levels are increasing.

Rapid systemic up-regulation of genes after heat-wounding and electrical stimulation.

When one leaf of a tomato plant is electrically-stimulated or heat-wounded, proteinase inhibitor genes are rapidly up-regulated in distant leaves. The identity of the systemic wound signal(s) is not yet known, but major candidates include hormones transmitted via the phloem or the xylem, the electrically-stimulated self-propagating electrical signal in the phloem (the action potential, AP), or the heat-wound-induced surge in hydraulic pressure in the xylem evoking a local change in membrane potential in adjacent living cells (the variation potential, VP). In order to discriminate between these signals we have adopted two approaches. The first approach involves applying stimuli that evoke known signals and determining whether these signals have similar effects on the "model" transcripts for proteinase inhibitors (pin) and calmodulin (cal). Here we show that a heat wound almost invariably evokes a VP, while an electrical stimulation occasionally evokes an AP, and both of these signals induce accumulation of transcripts encoding proteinase inhibitors. The second approach involves identifying the array of genes turned on by heat-wounding. To this end, we have constructed a subtractive library for heat-wounded tissue, isolated over 800 putatively up-regulated clones, and shown that all but two of the fifty that we have analyzed by Northern hybridization are, indeed, up-regulated. Here we show the early kinetics of up-regulation of three of these transcripts in the terminal (4th) leaf in response to heat-wounding the 3rd leaf, about 5 cm away. Even though these transcripts show somewhat different time courses of induction, with one peaking at 30 min, another at 15 min, and another at 5 min after flaming of a distant leaf, they all exhibit a similar pattern, i.e., a transient period of transcript accumulation preceding a period of transcript decrease, followed by a second period of transcript accumulation.

Is membrane potential involved in calmodulin gene expression after external stimulation in plants?

In Bidens pilosa (cv. radiata), a non-injurious stimulus induces a local and transient change in membrane potential, and an injurious stimulus induces a transmitted electrical signal described as the combination of an action potential and a slow wave. We have studied calmodulin gene expression after these stimuli. When the stimulus is non-injurious, calmodulin mRNA accumulation is only increased in the stimulated region. In contrast, when the stimulus is injurious, mRNA accumulation takes place in both wounded and distant, unwounded tissue. We propose that the slow wave plays a role in the long-distance transmission of a wound-induced information in plants.

Risk factors in community-acquired chronic hepatitis C virus infection: a case-control study in Italy.

AIMS/METHODS: A case-control study was carried out in Italy to assess the risk factors associated with chronic hepatitis C virus infection. Five hundred consecutive chronic anti-hepatitis C virus positive cases and 500 sex and exactly age-matched anti-hepatitis C virus negative/HBsAg negative controls entered the study. Information was collected through an interviewer-administered questionnaire. The adjusted Odds Ratios linking hepatitis C virus infection and risk factors were estimated by conditional multiple logistic regression. Demographic and socio-economic characteristics were similar in cases and controls. Seventy-five percent of patients were aged over 40: males were prominent in the group < or = 40, while the number of females increased with age. RESULTS: As expected, drug addiction and blood transfusion emerged as independent risk factors: blood transfusion in all ages and in both sexes, drug addiction only in subjects under 41 years and mostly in males. Other risk factors independently associated with hepatitis C virus infection were: previous use of non-disposable needles, previous tuberculosis, and prolonged hospitalization before 1970. A history of sexually-transmitted diseases was not associated. CONCLUSIONS: This study shows that the great spread of hepatitis C virus in Italy may have occurred several years ago through parenteral routes which are not now operating. Modern hygienic and sanitation measures have significantly controlled exposure to the infection, which in the younger generations is confined to high-risk groups such as drug addicts.

Cloning of the pyr4 gene encoding orotidine-5'-phosphate decarboxylase in Cephalosporium acremonium.

We have cloned the Cephalosporium acremonium pyr4 gene by cross-hybridization with the equivalent gene from Neurospora crassa, the closest relative from which this gene is available. The C. acremonium pyr4 gene complements an E. coli pyrF mutant lacking orotidine-5'-phosphate decarboxylase (OMPdecase), and most probably does not contain introns. Maxicell analysis in E. coli shows that it encodes a 46 kDa polypeptide. The C. acremonium OMPdecase contains a highly conserved pentadecapeptide characteristic for this category of enzyme. Extensive sequence comparison suggests an important role of this region in enzymatic activity.