Detection of hepatitis A virus genotype IB variants in clams from Maputo Bay, Mozambique

Clams provide an important source of food and income for the population of Maputo, Mozambique, where conditions of poor water supply and inadequate sanitation favor endemic infection with hepatitis A virus (HAV). To determine the role of bivalves in an endemic area, clams gathered from Maputo Bay were bought from market and examined for HAV. Four batches, total 150 clams, were sampled over the year. RNA extracted from individual digestive glands was assayed by nested RT-PCR and sequencing of HAV 5' noncoding region (5' NCR). Specific HAV signals were detected in one batch, 23 of 34 clams (67%) testing positive. Phylogenetic analyses of VP3/VP1, VP1/P2A, and 5' NCR determined clustering of clam strains as genotype I, subtype B. In addition to identifying HAV IB strains with predicted conserved amino acid sequence, IB variants exhibiting novel amino acid substitutions at the VP1/P2A junction were detected. HAV strains from clams showed 93%-99% homology with wild-type IB strains from South African outbreaks and from a panel of HAV IgM positive Swedish patients. DNA from enteric human adenovirus 40/41 was found in a limited number of clams from two batches, 6/34 (17%) and 4/35 (11%). Detection of HAV subgenotype IB in bivalves provided indirect evidence of the strains circulating in a densely populated coastal region where HAV is presumed to be hyperendemic. The results suggest that clams may be an important source of HAV in Maputo region, and indicate the need for further molecular study of strains circulating in the indigenous population.     

Lymphocyte lycopene concentration and DNA protection from oxidative damage is increased in women after a short period of tomato consumption

Several epidemiologic studies have suggested a role of tomato products in protecting against cancer and chronic diseases. In nine adult women, we evaluated whether the consumption of 25 g tomato puree (containing 7 mg lycopene and 0.3 mg beta-carotene) for 14 consecutive days increased plasma and lymphocyte carotenoid concentration and whether this was related to an improvement in lymphocyte resistance to an oxidative stress (500 micromol/L hydrogen peroxide for 5 min). Before and after the period of tomato intake, carotenoid concentrations were analyzed by HPLC and lymphocyte resistance to oxidative stress by the Comet assay, which detects DNA strand breaks. Intake of tomato puree increased plasma (P <0.001) and lymphocyte (P<0.005) lycopene concentration and reduced lymphocyte DNA damage by approximately 50% (P<0.0001). Beta-carotene concentration increased in plasma (P<0.05) but not in lymphocytes after tomato puree consumption. An inverse relationship was found between plasma lycopene concentration (r = -0.82, P<0.0001) and lymphocyte lycopene concentration (r = -0.62, P<0.01) and the oxidative DNA damage. In conclusion, small amounts of tomato puree added to the diet over a short period can increase carotenoid concentrations and the resistance of lymphocytes to oxidative stress.     

The use of synthetic prosthesis in the treatment of hernia

The authors report their experience on synthetic prosthesis used in 36 patients with abdominal hernias. Thirty six cases of incisional hernias have been analyzed by Institute of General Surgery of Messina University from 1992 to 1998. Among the 33 cases with median parietal defect, in 7 cases concerning great incisional hernias group a PTEF-e prosthesis, which was positioned in an intraperitoneal site, was employed. In 21 cases of medium size incisional hernias they used a polypropylene prosthesis, which was positioned in an intraparietal or preperitoneal site. In 5 cases of small size incisional hernias, polypropylene prosthesis was placed in a superfascial site. In the 3 cases of medium size lateral laparocele++, polypropylene prosthesis was positioned in a intraparietal site. There was no operative mortality. Among postoperative complications, 4 seromas, 1 parietal and 1 subcutaneous haematoma and 2 haematomas in retroperitoneal site appeared. Clinical-instrumental controls, which have continued until two or three years after the operation were performed which pointed out a recurrence in just one case where the prosthesis was positioned in a superfascial site. To conclude, we consider that nowadays the use of synthetic prosthesis lets us face and solve problems concerning surgical treatment of incisional hernias with confidence.     

Composition and turnover of contractile proteins in volume-overtrained skeletal muscle

The purpose of this study was to find the composition shift of myosin heavy chain (MyHC) isoforms in overtraining in fast- and slow-twitch skeletal muscles and different changes in MyHC isofom composition, synthesis and turnover rate between 4-week and 6-week overtraining. Male Wistar rats were randomly assigned to 4-week and 6-week endurance training, 4-week and 6-week overtraining groups. Plantaris (Pla), extensor digitorum longus (EDL), and soleus (Sol) muscles were studied. Daily excretion of 3-methylhistidine (3-MeHis) pool as an indicator for protein degradation increased in the 4-week and 6-week overtraining group to 4.04 +/- 0.21 and 4.32 +/- 0.23 %/day subsequently in comparison with the control group (2.16 +/- 14 %/day, p < 0.001). In Pla muscle MyHC I isoform synthesis rate was 33 200 +/- 2150 (after 6-week overtraining 27 100 +/- 1800, p < 0.05), IIa 32 600 +/- 2100; IId 27 300 +/- 1890 and IIb isoform 20 100 +/- 1600 (after 6-week overtraining 15 500 +/- 1400, p < 0.05) dpm/M leucine/min. Actin synthesis rate increased in fast-twitch muscles during 4- and 6-week overtraining, and in soleus muscle during 6-week overtraining. In EDL and Sol muscle MyHC isoform composition during 6-week overtraining did not change significantly. During the 6-week overtraining the relative content of MyHC I and IIb isoforms decreased and IIa and IId isoforms increased in Pla muscle. The initial increase of MyHC IIb isoform after 4-week overtraining shows the higher stability of this isoform in comparison with MyHC I isoform in fast-twitch muscles during high volume exercise.     

Immunochemical quantification of metallothioneins in marine mollusks: characterization of a metal exposure bioindicator

A recombinant metallothionein CgMT1, from the Pacific oyster Crassostrea gigas, was synthesized and used as antigen in the development of antibodies and a specific enzyme-linked immunosorbent assay (ELISA). The ELISA showed that rabbit anti-CgMT1 IgG reacted with purified CgMT1 and MTs from other marine bivalves, indicating its suitability as a reagent to quantify MTs and for monitoring of metal contamination in field animals. Application of this assay to tissues excised from metal exposed C. gigas, showed that MT induction reached a saturation level in gills that was not observed in digestive gland. Quantification of MTs in gills and digestive gland of field-collected C. gigas showed that the MT level depended on the metal concentrations at the collection sites and may have been influenced by salinity variations at estuarine sites. Oysters from metal-contaminated estuarine sites showed lower MT concentrations than those from nonestuarine contaminated sites.     

Characterization of a new PEPD allele causing prolidase deficiency in two unrelated patients: natural-occurrent mutations as a tool to investigate structure–function relationship

Prolidase deficiency (PD) is a rare autosomal recessive disorder characterized mainly by skin lesions of the legs and feet, mental retardation, and respiratory infections. Mutations at the PEPD locus, located on chromosome 19, are responsible for this disease. We identified a new PEPD allele in two unrelated Portuguese PD patients by analyses of reverse transcribed PCR-amplified cDNA. We used SSCP analysis of seven overlapping fragments spanning the entire coding region of the gene and detected abnormal SSCP bands in two of them: PD3 (nt 425–743) and PD4 (nt 661–973). Direct sequencing of the mutant cDNA and genomic DNA revealed a new homozygous 3-bp deletion (Y231del) in both cases. Transient expression in PD fibroblasts of wild-type and mutant prolidase cDNA confirmed reduced activity of the construct carrying the 3-bp deletion. The mutation results in a loss of prolidase activity in skin fibroblasts. Intracellular accumulation of Gly-Pro dipeptide in long-term cultured fibroblasts was detected by capillary electrophoresis. The mutation falls in the 2 domain of the pita bread structure proposed for E. coli and human prolidase by Bazan et al. on the bases of their sequence homology with E. coli methionine aminopeptidase. Taking into account the effects of the described mutations on stability and activity of the enzyme, we propose the identification of three different functional regions.     

Therapeutic reactance as a predictor of outcome in the treatment of chronic depression

This study examined whether reactance would negatively influence treatment outcome in 347 patients diagnosed with chronic forms of depression and treated at 9 sites with either Nefazodone, cognitive-behavioral analysis system of psychotherapy (CBASP), or combination therapy. Contrary to our hypotheses, reactance positively predicted treatment outcome in CBASP on 2 of 4 scales. These effects were independent of the therapeutic alliance, which also positively predicted outcome. Reactance did not predict outcome in the groups receiving medication alone or in combination with CBASP. The findings suggest that reactance may be an asset in psychotherapy among chronically depressed individuals and that reactant patients can benefit from directive psychotherapy when therapists flexibly respond to perturbations in the therapeutic relationship. Results support the importance of Aptitude * Treatment interactions in psychotherapy outcome. The direction and significance of such interactions may vary with different forms of psychopathology.