Hepatitis B and C in the hemodialysis unit of Tocantins,Brazil: serological and molecular profiles

A survey was conducted in the hemodialysis population of the state of Tocantins, Brazil, aiming to assess the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections, to analyze associated risk factors, and also to investigate these viruses genotypes distribution. During January and March 2001, all patients (n = 100) were interviewed at the unique dialysis unit in Tocantins. Blood samples were collected and serum samples were screened for HBV serological markers. Hepatitis B surface antigen positive samples were tested for HBV DNA. All samples were also tested for anti-HCV antibodies and HCV RNA. An overall prevalence of 45% was found for HBV infection (4% were HBsAg/anti-HBc positive, 2% were anti-HBc only and 39% had anti-HBc/anti-HBs markers). Concerning HCV infection, anti-HCV and HCV RNA were detected in 13% and 14% of the subjects, respectively. Three patients were HCV RNA positive and anti-HCV negative, resulting in an overall HCV prevalence of 16%. Univariate analysis of risk factors showed that only shift and length of tile on hemodialysis were associated with HBV and HCV positivity respectively. Among the four HBsAg-positive samples, HBV DNA was detected in three of them, which were identified as genotype A by restriction fragment length polymorphism (RFLP) analysis. All 14HCV RNA-positive samples were genotyped by INNO-LiPA. Genotypes la and 3a were found in 85% and 15%, respectively. The present data show low HBsAg and HCV prevalence rates. The risk factors associated with HBV and HCV positivity suggest that nosocomial transmission may influence in spreading these viruses in the dialysis unit studied.     

Development of a patient-specific anatomical foot model from structured light scan data

The use of anatomically accurate finite element (FE) models of the human foot in research studies has increased rapidly in recent years. Uses for FE foot models include advancing knowledge of orthotic design, shoe design, ankle–foot orthoses, pathomechanics, locomotion, plantar pressure, tissue mechanics, plantar fasciitis, joint stress and surgical interventions. Similar applications but for clinical use on a per-patient basis would also be on the rise if it were not for the high costs associated with developing patient-specific anatomical foot models. High costs arise primarily from the expense and challenges of acquiring anatomical data via magnetic resonance imaging (MRI) or computed tomography (CT) and reconstructing the three-dimensional models. The proposed solution morphs detailed anatomy from skin surface geometry and anatomical landmarks of a generic foot model (developed from CT or MRI) to surface geometry and anatomical landmarks acquired from an inexpensive structured light scan of a foot. The method yields a patient-specific anatomical foot model at a fraction of the cost of standard methods. Average error for bone surfaces was 2.53 mm for the six experiments completed. Highest accuracy occurred in the mid-foot and lowest in the forefoot due to the small, irregular bones of the toes. The method must be validated in the intended application to determine if the resulting errors are acceptable.     

Carvacrol Depends on Heme Oxygenase-1 (HO-1) to Exert Antioxidant,Anti-inflammatory,and Mitochondria-Related Protection in the Human Neuroblastoma SH-SY5Y Cells Line Exposed to Hydrogen Peroxide

Neurochemical Research - The link between mitochondrial dysfunction, redox impairment, and inflammation leads to increased rates of brain cells loss in neurodegenerative diseases and in affective...     

Paracoccidioides restrepiensis B339 (PS3) and P. lutzii LDR2 yeast cells and soluble components display in vitro hemolytic and hemagglutinating activities on human erythrocytes

Paracoccidioidomycosis (PCM) is a systemic mycosis caused by thermally dimorphic fungi of the Paracoccidioides species complex. Several pathogenic fungi produce hemagglutinins and hemolysins, which are virulence factors involved in adhesion of pathogens to host tissues or cells and in destruction of erythrocytes. The present research investigated hemolytic and hemagglutinating activities of yeast cells and soluble components from P. restrepiensis (PS3; formerly P. brasiliensis B339) and P. lutzii (LDR2). Different concentrations of live and heat‐killed yeast cells and soluble components from a cell free antigen preparation (native or heated, 56°C or 100°C, 30 min) were mixed with 1% human erythrocyte suspensions. Yeast cells from both species caused hemolysis, P. lutzii LDR2 being more strongly hemolytic than P. restrepiensis B339, whereas the opposite phenomena occurred with soluble components in most conditions. Live or heat‐killed yeast cells of both fungi agglutinated erythrocytes, but only heated soluble components from P. restrepiensis B339 showed hemagglutinating activity. In conclusion, yeast cells of P. restrepiensis B339 and P. lutzii LDR2 produce hemolysins and hemagglutinins, which are most likely predominantly restricted to yeast cells in P. lutzii LDR2 and predominantly released in soluble form by P. restrepiensis B339, requiring further study.

     

Impact of microbial growth inhibition and proteolytic activity on the stability of a new formulation containing a phytate-degrading enzyme obtained from mushroom

The development of stable enzymes is a key issue in both the food and feed industries. Consequently, the aim of the current study is to evaluate the impact of various additives (sodium chloride, sodium citrate, mannitol, methylparaben, polyethylene glycol 3350, ethylenediaminetetraacetic acid disodium salt, and a serine protease inhibitor) on the stability of a mushroom phytase produced by solid-state cultivation and recovery. Also observed was the effect of the additives on microbial growth inhibition by monitoring both the change in optical density over 30 days of storage and proteolytic activity. Initially, eight experimental formulations were prepared along with a control. After screening, a 3² factorial design was applied to define suitable concentrations of the selected additives. Among the eight formulations tested, the formulation containing NaCl, PEG 3350, and methylparaben retained all of the initial phytase activity after 50 days of storage, with no detected interference from protease activity. Sodium citrate, a metal chelation agent, presented the unusual effect of reducing protease activity in the formulations. Although all formulations presented better phytase stability when compared to the control, NaCl and PEG were both able to prolong the stability of the enzyme activity and also to inhibit microbial growth during storage, making them favorable for application as food and feed additives.     

Quantitative estimates of the distribution of homoserine dehydrogenase isozymes in maize tissues

The low molecular weight threonine-resistant (class I) and the higher molecular weight threonine-sensitive (class II/III) isozymes of homoserine dehydrogenase (EC 1.1.1.3) isolated from Zea mays L. were shown to differ in stability during incubations in the presence of urea. Class II/III was inactivated by urea in a time- and concentration-dependent manner, with complete inactivation occurring within 24 hours at 5 degrees C in 4.0 m urea. Under identical conditions, neither the activity nor the properties of class I were affected. Therefore, it was possible to estimate the amounts and properties of both maize isozymes in crude mixtures by measurements of enzyme activity before and after treatment with urea.The relative amounts of the two isozymes proved to be tissue-specific. When shoots of etiolated seedlings were extracted under optimum conditions, the resultant preparations contained about 16% class I and 84% class II/III. This distribution of isozymes, as well as the regulatory properties of class II/III, were constant during growth of the seedlings between 4 and 13 days. Enzyme preparations isolated from shoots of light-grown plants contained higher proportions of class I. The two isozymes were not uniformly distributed within leaves, as the basal meristematic region contained high levels of II/III and small amounts of I. During leaf maturation, the amount of II/III declined while the level of I remained constant or increased slightly. As a result, nearly half of the enzyme extracted from leaf tips was class I. The synthesis of specific members of the aspartate family of amino acids might be expected to differ when the ratio of threonine-sensitive to threonine-resistant homoserine dehydrogenase is altered. However, additional information on the subcellular localization and the catalytic characteristics of the two enzymes is required for evaluation of this possibility.     

Effects of plant age and extraction conditions on the properties of homoserine dehydrogenase isolated from maize seedlings

Homoserine dehydrogenase (EC 1.1.1.3) was extracted from shoots of etiolated seedlings of Zea mays L. which had been grown for periods ranging from three to thirteen days. Both the amount of enzyme extracted and its regulatory properties, as measured by the sensitivity of the enzyme to inhibition by the feedback modulator, l-threonine, were found to be a function of seedling age and extraction conditions. Equivalent amounts of enzyme with similar properties could be isolated from young seedlings under a variety of conditions. Extraction media containing comparatively low concentrations of the buffer component and a high concentration of dithioerythritol were found to be required for optimum extraction of the enzyme from shoots of seedlings grown longer than four days and from leaves of light-grown plants. In the absence of dithioerythritol, diminished regulatory control was observed to be a direct function of seedling age. Evidence of rapid desensitization of the enzyme during extraction was obtained from experiments in which dithioerythritol was added to extracts prepared in the absence of a thiol compound. Therefore, previous observations of growth-dependent desensitization in a number of plants could be due to incomplete extraction or to changes in cellular factors which inactivate and/or alter the enzyme. Whether the enzyme itself becomes increasingly susceptible to alteration during seedling growth remains to be established.     

The effect of ischaemia and reperfusion on sarcolemmal inositol phospholipid and cytosolic inositol phosphate metabolism in the isolated perfused rat heart

In this study the mass of polyphosphoinositides as well as the turnover of [³H]inositol phospholipids and [³H]inositol phosphates during ischaemia and short periods of reperfusion were studied in the isolated perfused rat heart. Since the phosphoinositides located within the sarcolemma are precursors for release of inositoltrisphosphate (InsP₃) and diacylglycerol, sarcolemmal membranes (rather than whole tissue) isolated at the end of the experimental procedure, were used. Hearts were prelabelled with [³H]inositol and subsequently perfused with 10 mM LiCI to block the phosphatidylinositol (PI) pathway. The results showed that 20 min of global ischaemia depressed the amount of [³H]inositol present in both sarcolemmal phosphatidylinositol-4-phosphate (PI-4-P) and phosphatidylinositol-4,5-bisphosphate (PI-4,5-P₂), as well as in the cytosolic [³H]inositol phosphates, [³H]InsP₂ and [³H]InsP₃. The mass of the sarcolemmal inositol phospholipids remained unchanged during ischaemia. Reperfusion caused an immediate (within 30 sec) increase in the amount of [³H]inositol in sarcolemmal PI, PI-4-P and PI-4,5-P₂. PI-4-P levels showed a transient increase after 30 seconds postischaemic reperfusion, while the mass of the other sarcolemmal inositol phospholipids, PI and PI-4,5-P₂, remained unchanged. [³H]Insp, [³H]InsP₂ and [³H]InsP₃ also increased significantly in comparison to ischaemic hearts after only 30 sec postischaemic reperfusion.In summary, the results obtained indicate inhibition of the PI pathway during ischaemia with an immediate significant stimulation upon reperfusion. In view of the capacity of InsP3 to mobilize Ca²⁺ the possibility exists that stimulation of this pathway during reperfusion may play a role in the intracellular Ca²⁺ overload, characteristic of postischaemic reperfusion.     

Unaltered Nodulation Competitiveness of a Strain of Bradyrhizobium sp. (Lotus) after a Decade in Soil

A Bradyrhizobium sp. (Lotus) strain that formed a soil population that was highly competitive for nodulation of Lotus pedunculatus 11 years after its introduction into a field soil and a culture of the same strain stored lyophilized were compared with an antibiotic-resistant mutant in respect of their nodulation competitiveness. The mutant was less competitive than the wild-type strain it was isolated from and had to be present at a cell ratio of 5.76:1 in mixed inoculum in sand culture to form 50% of the nodules on L. pedunculatus (50% nodulation value, 5.76). The 50% nodulation values for a soil population of the mutant mixed with soil populations of the lyophilized and field soil strain were, respectively, 6.83 and 5.77, indicating that the field soil strain was not significantly different from the lyophilized strain in nodulation competitiveness. A 50% nodulation value of 11.18 obtained when soil containing a recently established mutant population was mixed with the field soil containing the population established 11 years before, indicating that the plant infection technique underestimated cell numbers of the field soil population by 100%. Nodulation competitiveness was unaffected by the size of the strain populations in the range of 100 to 1,000 cells per g of soil; at 10 cells per g a significant correlation between strain ratios in nodules and in soil was still evident. The results indicated that apparently superior nodulation competitiveness of a well-established soil population relative to that of a subsequently introduced strain may not necessarily reflect the intrinsic competitive abilites of the strain(s) involved. The soil strain did not differ from laboratory-maintained cultures in antigenic properties, effectiveness, or whole cell protein electrophoresis profiles.