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To determine the efficacy and safety of the Ologen collagen matrix adjunctive to Ahmed valve surgery. A randomized prospective multicentre clinical trial involving 58 patients that were followed for one year. Conventional surgery with Ahmed valve was performed in 31 eyes (Control group/CG) and in 27 Ologen (Ologen group/OG) was placed over the valve's plate. Baseline data: age, corneal thickness, intraocular pressure(IOP) and antiglaucoma medications.Postoperative data (days 1, 7 and months 1, 3, 6 and 12): IOP, antiglaucoma medications, visual acuity and complications were recorded. Frequency of hypertensive phase, complete and qualified success and survival rate were studied. No differences were found between CG and OG in the baseline data. The only difference between groups was a significantly lower IOP at day 1. No other differences were found in the follow-up between groups. Hypertensive phase (56%CG and 55%OG, p = 0,947), complete success 28,6%CG and 30,4%OG (p = 0,88) and qualified success 96,4% and 95,9%(p = 0,794). Survival rates at 1 year were 76,7%(CG) and 69,2%(OG)(p = 0,531). 38,7% of patients in the CG suffered some complication during follow-up and 61,5% in OG(p = 0,086). Ologen does not increase safety or efficacy in Ahmed valve surgery at one-year follow-up. This is the first study that shows no benefit of Ologen adjunctive to this surgery.

Strains of Schizosaccharomyces pombe are being increasingly investigated with regards to their grape winemaking potential either in combination with the typical production yeast, Saccharomyces cerevisiae, or in monoseptic fermentations. Their ethanol tolerance and ability to degrade L-malic acid is oenologically convenient but contrasts with the comparatively high acetic acid and acetaldehyde formation potential which is considered undesirable, especially in white winemaking. The purpose of this work was to investigate the performance of a selected S. pombe strain in monoseptic femerntations of white grape must. Traditional batch fermentations were compared with an innovative and automated fed-batch fermentation technique were sugar concentrations are kept low during fermentations to decrease sugar induced osmotic stress. Because of its known effect on growth and ethanol tolerance, the effect of Mg was also tested. While Mg supplementation was not shown to significantly influence residual values of sugars, ethanol, glycerol, organic acids and acetaldehyde, the application of the fed-batch technique led to a fundamental change in yeast physiology. While glycerol values were only slightly reduced, the fed-batch approach allowed obtaining wines devoid of acetic acid whose levels were considerable in wines produced by the traditional batch technique (0.6 g/L). The work demonstrates that the acetic acid metabolism of S. pombe is associated to sugar induced osmotic stress such as for S. cerevisiae, too, and may be controlled by application of suitable fermentation techniques for winemaking., (Crown Copyright © 2018. Published by Elsevier Ltd. All rights reserved.)

Aims:   To identify genes and proteins involved in adaptation to low-temperature fermentations in a commercial wine yeast., Methods and Results:   Nine proteins were identified as representing the most significant changes in proteomic maps during the first 24 h of fermentation at low (13°C) and standard temperature (25°C). These proteins were mainly involved in stress response and in glucose and nitrogen metabolism. Transcription analysis of the genes encoding most of these proteins within the same time frame of wine fermentation presented a good correlation with proteomic data. Knockout and overexpressing strains of some of these genes were constructed and tested to evaluate their ability to start the fermentation process. The strain overexpressing ILV5 improved its fermentation activity in the first hours of fermentation. This strain showed a quicker process of mitochondrial degeneration, an altered intracellular amino acid profile and laxer nitrogen catabolite repression regulation., Conclusions:   The proteomic and transcriptomic analysis is useful to detect key molecular adaptation mechanisms of biotechnological interest for industrial processes. ILV5 gene seems to be important in wine yeast adaptation to low-temperature fermentation., Significance and Impact of the Study:   This study provides information that might help improve the future performance of wine yeast, either by genetic modification or by adaptation during industrial production., (© 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.)

Aims: For this study, we performed a genetic screen of S. cerevisiae's deletion library for mutants sensitive to dehydration stress, with which we aimed to discover cell dehydration-tolerant genes., Methods and Results: We used a yeast gene deletion set (YGDS) of 4850 viable mutant haploid strains to perform a genome-wide screen for the identification of desiccation stress modifiers. SIP18 is among the genes identified as essential for cells surviving to drying/rehydration process. Deletion of SIP18 promotes the accumulation of reactive oxygen species and enhances apoptotic and necrotic cell death in response to dehydration/rehydration process., Conclusions: SIP18p acts as an inhibitor of apoptosis in yeast under dehydration stress, as suggested by its antioxidative capacity through the ROS accumulation reduction after an H(2) O(2) attack., Significance and Impact of the Study: To our knowledge, this is the first systematic screen for the identification of putative genes essential to overcoming cell dehydration process. A broad range of identified genes could help to understand why some strains of high biotechnological interest cannot cope with the drying and rehydration treatments. Dehydration sensitivity makes these strains not suitable to be commercialized by yeast manufactures., (© 2011 No claim to Spanish Government works Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.)

We evaluated the effect of Ascaris lumbricoides on Giardia duodenalis infection and TH1/TH2 type immune mechanisms toward this parasite in 251 rural parasitized and 70 urban non-parasitized school children. The children were classified according to light (0-5000 eggs/g faeces) or moderate (>5001-50,000 eggs/g faeces) A. lumbricoides infection. Anti G. duodenalis skin hyper-reactivity, IgE, IgG, IL-13, IFN γ, IL6 and IL-10 levels were compared among G. duodenalis infected and non-infected children according to light or moderate A. lumbricoides infection. It was found that 62% of the A. lumbricoides moderately infected children were co-infected by G. duodenalis compared to 45% of the lightly infected group. After treatment, 42% of the A. lumbricoides moderately group were infected with G. duodenalis compared to 11% of their lightly counterparts, being A. lumbricoides IL-10 levels higher (p<0.0001) in the moderately infected group. In the A. lumbricoides lightly parasitized children, G. duodenalis infection was associated to a significant increase (p<0.005) of the levels of G. duodenalis IL-13, IFN-γ, IL-6, IgE, IgG and skin test hyper reactivity. In contrast, there was no effect of G. duodenalis infection in the elevation of these parameters among the A. lumbricoides moderately parasitized group, being those levels similarly lower as those observed in the control group. Inverse correlations were found between the levels of anti G duodenalis antibodies, skin test hyper-reactivity and cytokines with the intensity of A. lumbricoides infection (p>0.0001) and A. lumbricoides IL-10 levels (p>0.0001), suggesting that co-infection with A. lumbricoides may affect both TH1 and TH2 type immunity against G. duodenalis that may play an important role in the susceptibility to the infection after chemotherapy in children from endemic areas., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Namaqua rock mice (Aethomys namaquensis) consume nectar xylose when visiting Protea flowers. Whole-animal metabolism studies suggest that the gastrointestinal microflora plays an important role in xylose metabolism in A. namaquensis. We collected caecal contents under anaerobic conditions, cultured caecal microflora both aerobically and anaerobically, and assessed caecal microbial xylose utilization using a (14)C-xylose incubation assay. All four mice sampled hosted culturable caecal micro-organisms that tested positive for xylose utilization. These were classified by 16S rRNA based taxonomy as: Bacillus subtilis, Bacillus pumilus, Bacillus licheniformis, Shigella boydii, Arthrobacter sp. and members of the fungal genera Aspergillus and Penicillium. Cultures of these isolates were then analyzed by gas chromatography to determine the types and quantities of short-chain fatty acids produced by xylose fermentation. These results are discussed in the context of other studies of gut microflora in vertebrates.

Background: The clinical implementation and evaluation of heroin substitution programmes have been confounded by the lack of objective and validated biomarkers for illicit heroin use in patients prescribed pharmaceutical heroin. This study examined the capacity to detect illicit heroin use by gas chromatography-mass spectrometry (GC-MS) analysis of urine samples for the presence of opium impurities common to illicit, but not pharmaceutical heroin., Aims: To characterize the diagnostic properties of the metabolites of noscapine and papaverine in comparison to morphine as a gold-standard marker of illicit heroin use; and to examine the relationships between the self-reported time since most recent heroin use and the detection of these opioids in urine., Design: A cross-sectional study of 52 opioid-dependent patients in treatment (not prescribed heroin), who self-reported illicit heroin use within the preceding 2 weeks. Self-report data regarding recent drug use and a urine sample were collected. GC-MS analyses of urines were conducted and reported by laboratory staff blinded to self-report data., Findings: The metabolites of papaverine (hydroxypapaverine and dihydroxypapeverine) were found to have high sensitivity, specificity and negative predictive values as markers for illicit heroin use compared to the 'gold-standard' morphine. Other opioids, including 6-mono-acetylmorphine (6-MAM), codeine and noscapine metabolites (e.g. meconine) were less adequate in detecting heroin use., Conclusions: GC-MS detection of papaverine metabolites in urine appears to be suitable method of identifying illicit heroin use for clinical and research purposes.

To illustrate the effect of a cellulose-binding domain (CBD) on the enzymatic characteristics of non-cellulolytic exoglucanases, 10 different recombinant enzymes were constructed combining the Saccharomyces cerevisiae exoglucanases, EXG1 and SSG1, with the CBD2 from the Trichoderma reesei cellobiohydrolase, CBH2, and a linker peptide. The enzymatic activity of the recombinant enzymes increased with the CBD copy number. The recombinant enzymes, CBD2-CBD2-L-EXG1 and CBD2-CBD2-SSG1, exhibited the highest cellobiohydrolase activity (17.5 and 16.3 U mg(-1) respectively) on Avicel cellulose, which is approximately 1.5- to 2-fold higher than the native enzymes. The molecular organisation of CBD in these recombinant enzymes enhanced substrate affinity, molecular flexibility and synergistic activity, contributing to their elevated action on the recalcitrant substrates as characterised by adsorption, kinetics, thermostability and scanning electron microscopic analysis.

A previously described method for the analysis of hair has been modified to include analysis for amphetamines, benzodiazepines, cocaine and its metabolites, methadone and its metabolite, and phencyclidine in addition to opiates on a sample of hair. The samples of hair were washed twice with dichloromethane and cut into 1-mm segments prior to extraction with methanol at 45 degrees C for 18 h. The extracts were split into two parts; both were evaporated to dryness. One half of the extract was derivatized using MBTFA for analysis of amphetamines, and the other half was derivatized using MTBSTFA for analysis of the remaining drugs. The extracts were analyzed using electron impact gas chromatography-mass spectrometry operating in selected ion monitoring mode. In total, 18 drugs of abuse/metabolites could be detected. The method was used to screen 20 hair samples from patients attending a methadone-maintenance clinic.

A method for the simultaneous analysis of urine for the major drugs of abuse is described. The analytical procedure uses solid-phase extraction (SPE), gas chromatography-mass spectrometry (GC-MS) and a semi-automated identification process. It allows simultaneous extraction, derivatization and analysis of acidic, neutral and basic drugs from urine. Urine samples were subjected to enzymatic hydrolysis followed by SPE using Bakerbond narc-2 columns. The eluant was selectively derivatized with N-methyl-bis-trifluoroacetamide (MBTFA) and N-methyl-N-trimethylsilyltrifluoroacetamide + 1% trimethylchlorosilane. Analysis was performed using a GC-MS system operating in full scan mode. A simple macro programme was written to enhance the mass spectra identification capabilities of the MS software by producing extracted ion chromatograms (EIC) for the drugs of interest. Once a suspect compound was indicated by EIC, the mass spectrum of the compound was searched manually against reference libraries for positive identification and the retention time checked against that of the standard. This procedure has increased both the amount and the reliability of information given to clinicians without increasing the cost per sample. The system has been in routine operation for 24 months, processing up to 40 urine samples per day, with a usual turn-around time of 48 h.

The formation of DNA adducts by the covalent binding of genotoxic chemicals to DNA represents a valuable marker for assessing exposure to carcinogens but as yet the role of DNA adducts as a biomarker of carcinogenic susceptibility still needs to be clearly ascertained. To address this question an animal study was instigated using mice (SWR (high), BALB/c (intermediate) and C57BL/6J (low)) varying in their susceptibility to lung carcinogenesis. Groups of animals from each strain were dosed with a single intraperitoneal injection of saline or N -nitrosodiethylamine (NDEA) at 15 or 90 mg kg(-1) body weight. Lung and liver tissues were removed at different time points following dosing. Further groups of mice dosed with the same regime had urine samples collected 24 h post dosing and were then left up to 18 months to allow for the development of tumours. Immunoslot-blot analysis was used for the determination of N-7 ethylguanine (N-7EtG) and O(6) ethylguanine (O(6)EtG) adduct levels in the DNA from the tissues and gas chromatography-mass spectrometry (GC-MS) was used to determine N-3 ethyladenine (N-3EtA) adduct levels in the urine samples. Levels of alkyltransferase (ATase) were also determined in the tissues. The results showed that the DNA adduct levels and persistence were similar across the three strains of mice following dosing with 15 and 90 mg kg(-1) NDEA. High levels of adducts were observed in the urine of the BALB/c strain, implying an increased metabolic or repair capacity in this strain. However there were no differences in the levels of ATase in the lung and liver of the three strains of mice following dosing with 15 mg kg(-1) NDEA. The incidence of tumours in C57BL/6J mice was lower compared with the other two strains and showed a dose dependent increase. The results from this study show that the differences in susceptibility to lung carcinogenesis between the three strains of mice do not appear to be linked to the formation of the two adducts detected. These results imply that dosing with NDEA resulted in toxicity which may have led to cell death and induction of tumours by compensatory cell proliferation. Although these results do not allow decisive conclusions to be drawn concerning the relationship between total levels of DNA adducts and differences in carcinogenic susceptibility for the three strains of mice it is clear that the increased presence of a DNA adduct in the target tissue increases the likelihood of tumour development.

We measured the levels of thyroxine-binding prealbumin (TBPA), C-reactive protein (CRP), and interleukin-6 (IL-6) in sera from protein-energy undernourished and control children either without or with associated clinical infections. Levels of TBPA were significantly lower in undernourished than in control children without clinical infections. Mean serum TBPA concentration was also significantly lower in both clinically infected undernourished and control children than in those of the same nutritional status but without overt infections. Significant positive correlations were observed between the concentration of TBPA and the Z-scores of weight for age, height for age, and weight for height in all children without clinical infections, which disappeared in clinically infected children. CRP and IL-6 increased to approximately similar levels in sera from undernourished and control children with overt infections. An inverse correlation between the levels of TBPA and CRP and a significant positive correlation between the concentration of CRP and IL-6 were observed in children with associated clinical infections. Serum IL-6 and TBPA showed a small negative relationship in patients with overt infections. Thus, TBPA is a sensitive marker of undernutrition only in the absence of clinical infections, and the capacity to increase production of IL-6 and CRP during infections is preserved in protein-energy undernutrition.

A ring-test was organised between three laboratories using different versions of the modified Edman degradation technique for the gas chromatographic-mass spectrometric determination of N-terminal valine adducts of styrene 7,8-oxide. The analyses were performed on a sample of human haemoglobin reacted in vitro with styrene 7,8-oxide and on a set of five haemoglobin samples from mice dosed by i.p. injection of styrene. Strong correlations between the haemoglobin adduct determinations of the different laboratories were observed. However, covariance analysis revealed different slopes for the dose-response curves, indicating differences for the calibration of the reference globin or reference peptide.

Hepatic and erythrocytic glutathione peroxidase activity, together with malondialdehyde levels, were determined as indicators of peroxidation in 83 patients from whom liver biopsies had been taken for diagnostic purposes. On histological study, the patients were classified into groups as minimal changes (including normal liver), steatosis, alcoholic hepatitis, hepatic cirrhosis, light to moderately active chronic hepatitis, and severe chronic active hepatitis. The glutathione peroxidase activity in erythrocytes showed no significant changes in any liver disease group. In the hepatic study, an increased activity was observed in steatosis with respect to the minimal changes group, this increased activity induced by the toxic agent in the initial stages of the alcoholic hepatic disease declining as the hepatic damage progressed. There was a negative correlation between the levels of hepatic malondialdehyde and hepatic glutathione peroxidase in subjects with minimal changes. This suggested the existence of an oxidative equilibrium in this group. This equilibrium is broken in the liver disease groups as was manifest in a positive correlation between malondialdehyde and glutathione peroxidase activity.

The yeast Yarrowia lipolytica was shown to be sensitive to the aminoglycoside antibiotic hygromycin B. Spontaneous resistants appeared at a frequency of (2-5) x 10(-7) in media containing 100 mg/l drug. In order to develop a new selective marker for the transformation of this yeast, we constructed new plasmids expressing the Escherichia coli hygromycin-resistance gene (hph) under the control of the promoter and terminator sequences of the strongly expressed XPR2 gene of Y. lipolytica. Direct selection of hygromycin-B-resistant transformants on complete medium was very efficient and resulted in transformation frequencies comparable to those observed with conventional auxotrophic markers. This new marker can be used for integrating single copies of plasmid and for gene disruption and provides a convenient tag for genetic studies.

Due to its short half life and high tryptophan content, prealbumin has been considered a sensitive indicator of protein and/or energy deficiency. In addition, prealbumin diminishes during the acute phase response elicited by either infection or tissue injury. Serum levels of prealbumin were determined in undernourished children with or without associated clinical infection and in their infected or non-infected controls matched for age, sex, race and socioeconomical conditions. Serum levels of prealbumin were significantly lower in undernourished than in control children without overt infections. On the other hand, the concentrations of serum prealbumin diminished significantly and to similar levels both in undernourished and control children with associated clinical infection as compared with those observed in non infected children belonging to the similar nutritional status. A positive correlation was found between prealbumin level and the Z-scores of weight-for-age, height-for-age and weight-for height in children without associated infections, which disappeared in clinically infected patients. Thus, prealbumin is a marker of undernutrition in the absence of infection and could be an earlier and more sensitive indicator of actual undernutrition caused by the metabolic effects of acute phase cytokines, than the anthropometrical measurements used here.

It is known that human hemoglobin contains low levels of N-terminal N-(2-hydroxyethyl)valine. Possible sources of this modified amino acid are exposure to ethylene oxide or other 2-hydroxy-ethylating agents. Although such processes are likely to occur endogenously, the exogenous contribution to the adduct formation is unclear. In order to explore the latter, we have analyzed N-(2-hydroxyethyl)valine in the globin of 49 pregnant women and evaluated the effect of smoking status, area of residence, and glutathione S-transferase M1 genotype on adduct levels. Transplacental transfer of hydroxyethylating agents was also studied by the analysis of umbilical cord hemoglobin. The adduct levels in smokers were significantly higher than those in nonsmokers. The adduct levels in umbilical cord blood globin were quantitatively related to those in maternal blood (maternal:fetal ratio 2.7 in smokers and 2.8 in nonsmokers). In the nonsmokers, there was no statistically significant difference in the adduct level between the urban and rural areas, but the level in suburbia tended to be lower than that in the rural area. In the combined smoker and nonsmoker groups, there was no effect of the glutathione S-transferase M1 genotype on levels of N-(2-hydroxyethyl)valine.

A case of a 55-year old male with Whipple's disease who consulted for dementia (aggressivity, memory loss of recent events, sleep rhythm disturbances) is presented. Furthermore, the patient had referred changes in intestinal movement four years beforehand in addition to erratic arthralgias. Response to treatment (trimethoprim-sulfamethoxazole for one year) was favorable with resolution of both the neurologic and gastrointestinal pictures. The radiologic alterations of this entity are reviewed and suggest that this disease should be considered as a cause of dementia.

Sixty women having hypertension during pregnancy were studied. A predominancy of age groups 20-24 years for a 28.3% was documented. The degree of participation of the multidisciplinary medical team is essential, as well as the close relation with the health area, since 94% of patients were recruited early and 80% received home visits by the family doctor and the nurse. There gas a predominancy of 23 cases with secondary school level for a 38%. Treatment was found to reduce arterial tension and satisfactory results were obtained for both the mother and the child.

This work is a very summarized Louis Pasteur's biography. As a scientist he discovered the dimorphism of the tartric acid crystals, the fermentation of several compounds (beer, wine, vinegar, butter) and how to avoid them thru the pasteurization, he also discovered the bacterial cause of many diseases of man and animals, changing the ways of human life. Thru sterilization, since him, the fatal outcome of many patients is avoided and surgery was made safe. He established bacteriology and immunology as branches of medicine and biology.

Isolated cerebral microvessels (ICMV) have been increasingly used to study microvascular metabolism and function. Despite this, little systematic information exists about their metabolic viability and energy status. To evaluate this, we determined the ATP content and ATP/ADP ratios of ICMV with an ultrasensitive bioluminometric assay which had been adapted for small samples. In calf cerebral microvessels, freshly isolated by a homogenization and sieving procedure, ATP content averaged 1.5 +/- 0.8 nmole/mg protein (mean +/- SE for 45 observations on 18 preparations.) The ATP/ADP ratio for these vessels was 0.96 +/- 0.4. Similar values for ATP were obtained in several preparations of ICMV from dog and rat brain. Values varied considerably in different preparations, probably due to variable degrees of damage incurred during isolation. When microvessel ATP concentrations and ATP/ADP ratios were low, they were dramatically improved by brief incubation (2 hr at 37 degrees) in an enriched tissue culture medium (Dulbecco's modification of Eagle's minimal essential medium, DMEM), or perhaps somewhat less effectively in a buffered saline solution (Earle's-Hepes) containing glucose. Boiling or incubation of microvessels with Triton X-100 lowered ATP values to less than 0.01 nmole/mg protein. The ATP content of our preparations of isolated microvessels was considerably higher than values previously reported by others using similar methods, but still less than that of cultured bovine vascular endothelial cells, even after correction for a 20% difference in intracellular water space.(ABSTRACT TRUNCATED AT 250 WORDS)

Glucose and beta-hydroxybutyrate metabolism were compared in isolated cerebral microvessels from chronically diabetic and hypoglycemic rats. As noted previously, glucose oxidation and conversion to lactate are diminished in rats with streptozotocin-induced diabetes. The decrease in glucose metabolism did not result from selective damage to diabetic vessels during isolation, since the ATP level and the ATP/ADP ratio were similar to those of nondiabetic rats, and O2 consumption was increased. In addition, cerebral microvessel oxidation of beta-hydroxybutyrate was enhanced by diabetes. By contrast, microvessels from rats made chronically hypoglycemic by insulinoma engrafting 30 days earlier had a more than twofold increase in glucose oxidation and conversion to lactate, whereas their oxidation of beta-hydroxybutyrate was diminished by 50%. Unlike the insulinoma rats, no consistent increase in glucose metabolism was observed in microvessels from rats made hypoglycemic either by acute insulin administration or by a 4-day infusion of insulin. These results indicate that diabetes, and under some circumstances chronic hypoglycemia, markedly alters fuel metabolism in the cerebral microvasculature.