Farming technology, biochemistry characterization, and protective effects of culinary-medicinal mushrooms Agaricus brasiliensis S. Wasser et al. and Lentinus edodes (Berk.) Singer: Five years of research in Brazil

The biotechnology, biochemical characterization, and protective effects of the himematsutake and shiitake mushrooms were studied for the Thematic Project, from 1998 to 2003. A new species, Agaricus Brasiliens is Wasser et al. (= A. blazei Murrill ss. Heinem.), was proposed for the cultivated lineages in Brazil. Interactions among lineages, substrates, casing layers, and fructification inductions (temperature alternations) significantly increased the productivity of A. brasiliensis in Brazil (from 40 g to 200 g of fresh mushroom kg -1 moist compost). However, pests and diseases (mainly Sciaridae flies and false truffle), drastically reduced the productivity of A. brasiliensis (below 50 g kg -1). Biochemically for each mushroom species, the polar extracts, no polars, and medium polarity presented the same organic substances; however, their concentrations differed as a result of the lineages, maturation of the fruit bodies, and cultivation conditions. In this aspect, concentration of linoleic acid in A. brasiliensis (added to animal food) was related directly to the chemical protection against carcinogenic drugs in mice. Aqueous extracts of Lentinus edodes (= Lentinula edodes (Berk.) Pegler) and A. brasiliensis may be preventive chemical protectors against mutagenic and carcinogenic drugs, depending on the lineage and extraction method (tea or juice). However, immunomodulator effects and tumor reduction were only observed with concentrated fractions (hexanic, methanolic, and ATF extracts). Aqueous extracts of A. brasiliensis and L. edodes have antibiotic-like substances and substances able to act as elicitors of resistance responses in some plants (local and systemic) and show a potential to be used in the alternative control of plant pathogens. © 2005 by Begell House, Inc.

Evaluating the karyotypic diversity in species of Hyla (Anura, Hylidae) with 2n = 30 chromosomes based on the analysis of ten species

Ten species of Hyla with 2n = 30 from Brazilian fauna were analysed cytogenetically. Hyla minuta is the unique presenting all bi-armed metacentric or submetacentric chromosomes in the karyotype, therefore, with the highest FN = 60. The remaining species have a variable number of uni-armed telocentric or subtelocentric chromosomes: H. cruzi, H. elianeae, and H. rubicundula with three pairs (FN = 54), H. berthalutzae, H. elegans, H. microps, and H. nana with four pairs (FN = 52), and H. nahdereri and H. sanborni with five pairs (FN = 50). The uni-armed elements are among pairs 5, 6, 7, 11, 14, and 15, which also appeared with metacentric or submetacentric morphology. The remaining chromosome pairs 1, 2, 3, 4, 8, 9, 10, 12, and 13 were never found to be telocentric or subtelocentric. AgNOR patterns are species-specific, the majority of the species exhibiting a single pair with AgNORs, with the exception of H. elegans and H. nana with more than one chromosome pair bearing this cytological marker. C banding was obtained in H. berthalutzae, H. cruzi, H. elegans, H. elianeae, H. microps, H. minuta, H. nahdereri, and H. nana, which showed positively stained centromeric heterochromatin. Our analysis confirms the great karyotypic diversity in the species of Hyla with 2n = 30, with no species sharing identical karyotypes.

Comparison of xylazine and medotomidine as premedicants for cats being anaesthetised with propofol-sevoflurane

The effects of premedicating cats with saline, xylazine or medetomidine before anaesthetising them with propofol-sevoflurane were compared. Twenty-four cats were randomly assigned to three groups of eight to receive either 0.25 ml of saline, 0.50 mg/kg of xylazine or 0.02 mg/kg of medetomidine intravenously, and anaesthesia was induced with propofol and maintained with sevoflurane. Medetomidine produced a greater reduction in the induction dose of propofol and fewer adverse postoperative effects than saline or xylazine. Hypoxaemia was observed after induction with propofol in the cats premedicated with saline and xylazine, but not in the cats given medetomidine. The cats treated with medetomidine and xylazine developed profound bradycardia. The blood pressure of the cats premedicated with saline and xylazine decreased, but the blood pressure of the cats premedicated with medetomidine was maintained. The cats premedicated with saline took longer to recover from anaesthesia than the other two groups.

Shear bond strength of adhesive systems to enamel and dentin. Thermocycling influence

Objectives: The purpose of the this study was to evaluate the influence of thermocycling on shear bond strength on bovine enamel and dentin surfaces of different adhesive systems. Methods: Thirty sound bovine incisors were sectioned in mesiodistal and inciso-cervical direction obtaining 60 incisal surfaces (enamel) and 60 cervical surfaces (dentin). Specimens were randomly assigned to 3 groups of equal size (n = 40), according to the adhesive system used: I-Single Bond; II-Prime & Bond NT/NRC; III-One Coat Bond. After 24-h storage in distilled water at 37 o C, each main group was divided into two subgroups: A- specimens tested after 24 h storage in distilled water at 37°C; B - specimens submitted to thermocycling (500 cycles). Shear bond strength tests were performed. Data were submitted to ANOVA and Tukey test. Results: Means (MPa) of different groups were: I-AE-16.96, AD-17.46; BE-21.60, BD-12.79; II-AE-17.20, AD-11.93; BE-20.67, BD-13.94; III-AE-25.66, AD-17.53; BE-24.20, BD-19.38. Significance: Thermocycling did not influence significantly the shear bond strength of the tested adhesive systems; enamel was the dental substrate that showed larger adhesive strength; One Coat Bond system showed the best adhesive strength averages regardless of substrate or thermocycling. © 2005 Springer Science + Business Media, Inc.

Effect of dietary supplementation with vitamin E and stocking density on macrophage recruitment and giant cell formation in the teleost fish, Piaractus mesopotamicus

The effect of dietary supplementation with 0, 100 and 450 mg of vitamin E (DL-α tocopheryl acetate)/kg of a dry diet on the kinetics of macrophage recruitment and giant cell formation in the pacu, maintained at different stocking densities (5 kg/m3 and 20 kg/m3), was investigated by insertion of round glass coverslips into the subcutaneous connective tissue. After a feeding period of 18 weeks, the coverslips were implanted and later removed for examination at 2, 7 and 15 days post-implantation. Fish fed diets supplemented with 450 mg of vitamin E showed an increase (P<0.05) in the accumulation of macrophages, foreign body giant cells and Langhans type cells. The kinetics of macrophage recruitment and giant cell formation on the glass coverslips appeared to be strongly influenced by vitamin E supplementation, since fish fed a basal diet and held at high stocking densities showed low numbers of adhering cells on the coverslips, and high concentrations of plasma corticosteroids. On the other hand, fish given a diet supplemented with 450 mg of vitamin E did not show a similar difference in plasma cortisol concentrations related to stocking density. The effect of cortisol concentrations on carbohydrate metabolism, analysed by assessment of plasma glycaemia, was not clear. Blood glucose concentrations did not vary substantially with the different treatments examined. These results suggest that vitamin E may contribute to the efficiency of the fish's inflammatory response by increasing macrophage recruitment and giant cell formation in the foreign body granulomatous reaction. Vitamin E appeared to act on the stress response of pacus by preventing a stress-related immunosuppression. © 2005 Elsevier Ltd. All rights reserved.

Evaluation of Strychnos pseudoquina St. Hil. leaves extract on gastrointestinal activity in mice

Strychnos pseudoquina St. Hil. (Loganiaceae) was investigated for its ability to protect the gastric mucosa against injuries caused by non-steroidal anti-inflammatory drugs (piroxicam) and a necrotizing agent (HCl/EtOH) in mice. The MeOH extract and enriched alkaloidic fraction (EAF) provided significant protection in experimental models wheer used at doses of 250 and 1000 mg/kg. In vivo tests were carried out to evaluate for possible toxic effects and no mortality was observed up to the 5 g/kg dose level. Phytochemical investigation led to the isolation of a new indole alkaloid, which elucidated the observed pharmacological effects. © 2005 Pharmaceutical Society of Japan.

Low ethanol consumption induces enhancement of insulin sensitivity in liver of normal rats

Moderate amounts of alcohol intake have been reported to have a protective effect on the cardiovascular system and this may involve enhanced insulin sensitivity. We established an animal model of increased insulin sensitivity by low ethanol consumption and here we investigated metabolic parameters and molecular mechanisms potentially involved in this phenomenon. For that, Wistar rats have received drinking water either without (control) or with 3% ethanol for four weeks. The effect of ethanol intake on insulin sensitivity was analyzed by insulin resistance index (HOMA-IR), intravenous insulin tolerance test (IVITT) and lipid profile. The role of liver was investigated by the analysis of insulin signaling pathway, GLUT2 gene expression and tissue glycogen content. Rats consuming 3% ethanol showed lower values of HOMA-IR and plasma free fatty acids (FFA) levels and higher hepatic glycogen content and glucose disappearance constant during the IVITT. Neither the phosphorylation of insulin receptor (IR) and insulin receptor substrate-1 (IRS-1), nor its association with phosphatidylinositol-3-kinase (PI3-kinase), was affected by ethanol. However, ethanol consumption enhanced liver IRS-2 and protein kinase B (Akt) phosphorylation (3 times, P < 0.05), which can be involved in the 2-fold increased (P < 0.05) hepatic glycogen content. The GLUT2 protein content was unchanged. Our findings point out that liver plays a role in enhanced insulin sensitivity induced by low ethanol consumption. © 2005 Elsevier Inc. All rights reserved.

Inhibitory effect of deferoxamine on Paracoccidioides brasiliensis survival in human monocytes: Reversal by holotransferrin not by apotransferrin

The mechanisms used by Paracoccidioides brasiliensis to survive into phagocytic cells are not clear. Cellular iron metabolism is of critical importance to the growth of several intracellular pathogens whose capacity to multiply in mononuclear phagocytes is dependent on the availability of intracellular iron. Thus, the objective of this paper was to investigate the role of intracellular iron in regulating the capacity of P. brasiliensis yeast cells to survive within human monocytes. Treatment of monocytes with deferoxamine, an iron chelator, suppressed the survival of yeasts in a concentration-dependent manner. The effect of deferoxamine was reversed by iron-saturated transferrin (holotransferrin) but not by nonsaturated transferrin (apotransferrin). These results strongly suggest that P. brasiliensis survival in human monocytes is iron dependent.

Cyclosporin but not tacrolimus significantly increases salivary cytokine contents in rats

Background: Cyclosporin (CsA) and tacrolimus (FK-506) are immunosuppressive drugs that specifically inhibit T-cell activation via calcineurin inhibition. Gingival overgrowth is a common side effect following the administration of CsA. The severity of gingival overgrowth seen in patients taking FK-506 is less than that observed with CsA. Little is known about the involvement of saliva in drug-induced gingival overgrowth. The purpose of this study was to investigate the salivary contents of tumor growth factor β1 (TGF-β1), epidermal growth factor (EGF), and interleukin-6 (IL-6) as well as the hystometry of gingival tissue obtained from rats treated with either FK-506 or CsA. Methods: For 30 or 60 days rats received daily subcutaneous injection doses of either CsA or FK-506 (10 mg/kg). The concentrations of TGF-β1, EGF, and IL-6 in saliva were determined by enzyme-linked immunosorbent assay, and after histological processing, the oral epithelium and connective tissue were assessed at the region of the lower first molars. Results: The levels of TGF-β1, EGF, and IL-6 in saliva were not significantly altered by any of the treatments after 30 days. After 60 days of treatment with CsA, gingival overgrowth and significant increase in salivary TGF-β1, EGF, and IL-6 concentrations were observed; no statistically significant changes were induced by FK-506. Conclusion: Within the limits of this experimental study, it can be concluded that CsA, but not FK-506, induced gingival overgrowth associated with an increase of the salivary levels of the cytokines TGF-β1, EGF, and IL-6.

β-carotene attenuates the paradoxical effect of tobacco smoke on the mortality of rats after experimental myocardial infarction

The objective of this study was to investigate the effects of exposure to tobacco smoke (ETS) in rats that were or were not supplemented with dietary β-carotene (BC), on ventricular remodeling and survival after myocardial infarction (Ml). Rats (n = 189) were allocated to 4 groups: the control group, n = 45; group BC administered 500 mg/kg diet, n = 49, BC supplemented rats; group ETS, n - 55, rats exposed to tobacco smoke; and group BC+ETS, n = 40. Wistar rats weighing 10O g were administered one of the treatments until they weighed 200 to 250 g (∼5 wk). The ETS rats were exposed to cigarette smoke for 30 min 4 times/d, in a chamber connected to a smoking device. After reaching a weight of 200-250 g, rats were subjected to experimental MI (coronary artery occlusion) and mortality rates were determined over the next 105 d. In addition, echocardiographic, isolated heart, morphometrical, and biochemical studies were performed. Mortality data were tested using Kaplan-Meyer curves and other data by 2-way ANOVA. Survival rates were greater in the ETS group (58.2%) than in the control (33.3%) (P = 0.001) and BC+ETS rats (30.0%) (P = 0.007). The groups did not differ in the other comparisons. Left ventricular end-diastolic diameter normalized to body weight was greater and maximal systolic pressures were lower in the ETS groups than in non-ETS groups. Previous exposure to tobacco smoke induced a process of cardiac remodeling after MI. There is a paradoxical protector effect with tobacco smoke exposure, characterized by lower mortality, which is offset by BC supplementation. © 2005 American Society for Nutritional Sciences.

Isolation of Mycobacterium gordonae from poultry slaughterhouse water in São Paulo State, Brazil

Water samples (24 untreated water, 12 treated water and 24 served water) used in different stages of the slaughter process were examined to identify a possible source of pathogenic mycobacteria. The isolates were identified based on microscopy, morphological and biochemical features, mycolic acid analysis and molecular method - PCR-restriction-enzyme analysis. Eighteen mycobacterial strains were isolated from 60 water samples: 11 from untreated water, 5 from treated water and 2 from served water. All mycobacteria isolated were identified as Mycobacterium gordonae and showed the following PRA genotypes: III (27.8%), IV (38.9%) and V (33.3%).

A comparison of mycolic acid analysis for nontuberculous mycobacteria identification by thin-layer chromatography and molecular methods

The development of fast, inexpensive, and reliable tests to identify nontuberculous mycobacteria (NTM) is needed. Studies have indicated that the conventional identification procedures, including biochemical assays, are imprecise. This study evaluated a proposed alternative identification method in which 83 NTM isolates, previously identified by conventional biochemical testing and in-house M. avium IS1245-PCR amplification, were submitted to the following tests: thin-layer chromatography (TLC) of mycolic acids and PCR-restriction enzyme analysis of hsp65 (PRA). High-performance liquid chromatography (HPLC) analysis of mycolic acids and Southern blot analysis for M. avium IS1245 were performed on the strains that evidenced discrepancies on either of the above tests. Sixty-eight out of 83 (82%) isolates were concordantly identified by the presence of IS1245 and PRA and by TLC mycolic acid analysis. Discrepant results were found between the phenotypic and molecular tests in 12/83 (14.4%) isolates. Most of these strains were isolated from non-sterile body sites and were most probably colonizing in the host tissue. While TLC patterns suggested the presence of polymycobacterial infection in 3/83 (3.6%) cultures, this was the case in only one HPLC-tested culture and in none of those tested by PRA. The results of this study indicated that, as a phenotypic identification procedure, TLC mycolic acid determination could be considered a relatively simple and cost-effective method for routine screening of NTM isolates in mycobacteriology laboratory practice with a potential for use in developing countries. Further positive evidence was that this method demonstrated general agreement on MAC and M. simiae identification, including in the mixed cultures that predominated in the isolates of the disseminated infections in the AIDS patients under study. In view of the fact that the same treatment regimen is recommended for infections caused by these two species, TLC mycolic acid analysis may be a useful identification tool wherever molecular methods are unaffordable.

Fermented soy product supplemented with isoflavones affected fat depots in juvenile rats

Objectives: This study investigated the effects of soy product fermented by Enterococcus faecium and Lactobacillus jugurti supplemented with isoflavones on adipose tissue, blood lipid, and glucose levels on juvenile rats. Methods: Rats were fed a cholesterol-enriched diet for 3 wk as a preliminary treatment to create hypercholesterolemia. They were then fed a chow diet (HC), a chow diet plus fermented soy product supplemented with isoflavones (HCFI), a chow diet plus placebo (HCP), or a chow diet plus placebo supplemented with isoflavones (HCPI), respectively, for an additional 3 wk. Results: The beneficial effects of fermented soy product supplemented with isoflavones on epididymal (EPI) and retroperitoneal (RET) fat pads was likely due to isoflavones because adipocyte circumference (micrometers) in the HC group was significantly larger (EPI: 105.66 ± 13.36; RET: 134.95 ± 25.40) than that in the HCFI group (EPI: 93.17 ± 12.80; RET: 108.62 ± 15.50) and HCPI group (EPI: 93.06 ± 15.10; RET: 112.34 ± 18.21). The probiotic micro-organism accentuated the antilipogenic effect of isoflavones on RET (HCFI: 108.62 ± 15.50 micrometers versus HCPI: 112.34 ± 18.21 micrometers). Moreover, the fermented product increased glucose concentration similar to that in the chow group but did not change blood lipids. Conclusion: This product may offer new approaches to obesity prevention. © 2005 Elsevier Inc. All rights reserved.

Skull and mandible formation in the cuckoo (Aves, Cuculidae): Contributions to the nomenclature in avian osteology and systematics

The study of the contributions of different bones to the formation of the skeleton in birds is necessary: (1) to establish homologies in comparative anatomy; (2) to delimit each bone structure correctly, mainly in relation to the skull and mandible where the bones are fused to each other in adults; and (3) to standardize nomenclature in avian osteology. In this paper at least one young specimen belonging to each sub-family of Cuculidae was examined in order to identify each bone in terms of boundaries and contributions to skull and mandible formation. These cuckoos specimens were also compared with adults and young of turacos and hoatzin. The results show little variation of skull and jaw among the young cuckoos studied compared with the variations among adult specimens. However, it provides new suggestions for the boundaries and nomenclature of certain osseous structures in the skull and mandible of birds, specifically fissura zona flexoria craniofacialis, prominetia frontoparietalis, crista temporalis transversa, processus squamosalis, fossa laterosphenoidalis, tuberculum laterosphenoidale and processus retroangularis. This study also provides more reliable homologies for use in cladistic analysis and above all it contributes to the phylogenetic position of Cuculidae within Neognathae, specifically the skull formation suggest that turacos and hoatzin are more similar to each other than either is to the cuckoos. © 2005 Taylor & Francis.

Age-age correlation for early selection of rubber tree genotypes in São Paulo State, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)