Location of the β-galactosidase of the yeast Kluyveromyces marxianus var. marxianus ATCC 10022

During the growth of Kluyveromyces marxianus var. marxianus ATCC 10022 on lactose, peaks of glucose, but not β-galactosidase activity, were detected in culture medium. Harvested and washed whole cells produced glucose and galactose from lactose, or ortho-nitro-phenol from the chromogenic substrate ortho-nitro-phenyl-β-D-galactopyranoside (ONPG), indicating that β-galactosidase is physically associated with cells. ONPG hydrolysis by whole cells presented a monophasic kinetics (Km 36.6 mM) in lactose exponential growth phase cells, but a biphasic kinetics (Km 0.2 and 36.6 mM) in stationary growth phase cells. Permeabilization with digitonin or disruption of cells from both growth phases led to monosite ONPG hydrolysis (Km 2.2 to 2.5 mM), indicating that β-galactosidase is not located in the periplasm. In addition, the energy inhibitors fluoride or arsenate, as well as the uncouplercarbonyl cyanide m-chlorophenylhydrazone (CCCP) prevented ONPG hydrolysis by whole cells. These findings indicate that energy coupled transmembrane transport is the rate-limiting step for intracellular ONPG cleavage. The taxonomic and physiologic implications of the exclusive intracellular location of β-galactosidase of K. marxianus var. marxianus ATCC 10022 are discussed. © 1996 Kluwer Academic Publishers.

Immunohistochemical aspects of basal cell adenoma and canalicular adenoma of salivary glands

Basal cell adenoma is a benign epithelial neoplasm with a uniform histologic appearance dominated by basaloid cells. Those cells may be distributed in various arrangements as solid, trabecular, tubular and membranous. Canalicular adenoma is also a benign neoplasm composed by columnar cells arranged in branching and interconnecting cords of single or double cell thick rows. There is some disagreement among investigators about whether canalicular adenoma should be included within the basal cell adenoma histologic spectrum. In the present study we compared the expression of cytokeratins (CK), vimentin and muscle-specific actin, utilizing immunohistochemical technique, in three cases diagnosed as basal cell adenomas predominantly of the solid type, and three cases of canalicular adenomas. The results obtained showed a distinct immunoprofile for both neoplasms. Solid areas of basal cell adenomas did not stain for any of the tested antibodies; only when there was tubular differentiation, those structures expressed CKs 7, 8, 14, and 19 in luminal cells and vimentin in non-luminal cells. On the other hand, canalicular adenomas strongly expressed CKs 7 and 13. The panel of antibodies utilized supports the separation of the two entities. © 2001 Elsevier Science Ltd.

Inhibin B and the ovarian follicular reserve

Inhibins are dimeric glycoproteins composed of an α-subunit and a βA-subunit (inhibin A) or βB-subunit (inhibin B), which inhibits pituitary gonadotropin secretion of FSH. The inhibin B is a product of the cohort of antral follicles. The ovarian follicle number decrease steadily as a function of increasing age, with consequent falls in the levels of inhibin B and increase of FSH levels. It is sufficient to maintain ovulatory function and continued secretion of estradiol. Elevated FSH levels seem to occur late in the sequence of events associated with ovarian failure. The inhibin B, produced by granulosa cells, is the earliest marker of the decline in ovarian follicular reserve across reproductive aging.

The ultrastructure of the pre-meiotic and meiotic stages of spermatogenesis in Plagioscion squamosissimus (Teleostei, Perciformes, Sciaenidae)

Spermatogenesis of 'corvina' P. squamosissimus starts from a stem cell that gives rise to germ cells. These cells are enveloped by Sertoli cells, forming cysts. The germ cells in the cysts are all at the same stage of development and are interconnected by cytoplasmic bridges. Spermatogonia are the largest germ cells. In the cysts, these cells differentiate into primary spermatogonia and secondary spermatogonia. The primary spermatogonia are isolated in the cyst and give rise to the secondary spermatogonia. After several mitotic divisions, they produce spermatocytes I, which can be identified by synaptonemal complexes in the nucleus. The spermatocytes I enter the first phase of meiosis to produce the spermatocytes II. These are not very frequently seen because they rapidly undergo a second phase of meiosis to produce spermatids.

Little cicada of sugarcane Mahanarva posticata (Homoptera: Cercopidae). A Brazilian agricultural pest. Morpho-histological study of salivary glands

Little cicadas are homopteran insect pests of sugarcane plantations. As these insects suck out the sap from the leaf parenchyma, they inoculate a toxic saliva that damages the plant vessels, thus promoting the loss of glucose by the affected plant. The morphological and histological analyses of the salivary glands of the little cicada Mahanarva posticata, revealed that these glands are formed by 2 portions: one portion comprises a group of acini and has been denominated as the principal gland; the second portion is filamentous in nature and has been denominated as the accessory gland; it is formed by very long and fine filaments. The acinous portion of the gland can be subdivided into 2 lobes: an anterior lobe formed by 3 lobules (I, II, III), and a posterior lobe formed by lobule IV and the excretory duct. Histologically, the salivary glands showed that the filaments are empty sutructures composed by several internal channels with secretion granules being observed in the cytoplasm of the cells of the secretory filaments. Lobules I and II of the principal gland are characterized by being highly basophilic and for accumulating a large amount of secretion in both the cytoplasm of the cells and inside secretion vesicles. Histochemically, we verified that the secretion produced by these glands is lipidic and protein in nature, with the production of polysaccharides being very low. The differences in stain and appearance of the different regions of the salivary gland lead us to believe that the final glandular product is lipoproteic in nature.

Interação das enterotoxinas estafilocócicas com o sistema imune do hospedeiro

Staphylococcal enterotoxins are among the most common etiologic agents that cause food poisoning and, possibly, nonmenstrual toxic shock syndrome. These enterotoxins are also called superantigens because they are potent T cell and macrophages activators. The superantigens bind directly to the major histocompatibility complex class II molecules on antigen-presenting cells and stimulate T cells expressing specific Vβ elements in the cell receptors. Excessive production of cytokines by these cells and macrophages are responsible for the pathogenesis of food poisoning. These cytokine include tumor necrosis factor (TNF)-α, interferon (IFN)-γ and interleukin (IL)-1, proinflamatory mediators with potent immunoenhancing effects; the nitric oxide (NO). It still has both effects citotoxic and regulatory roles in immune function.

Toxic and genotoxic effects of trivalent and hexavalent chromium - A review

During the last years, the emission of heavy metals to the environment has increased, causing a severe negative impact to the ecosystems and seriously compromising human health due to their mutagenic potential. Tri- (III) and hexavalent (VI) chromium (Cr) constitute the oxidative states of the metal chromium that are active in living organisms. These two oxidation states of the chromium differ with regards to their cellular effects, mainly due to the different abilities they possess in relation to easy of transport through biological membranes. Cr VI is transported into the cell through transference channels of endogenous anions that are isostructural and isoelectronical to Cr VI, such as SO 4 -2 and HPO 4 -2. On the other hand, Cr III is unable to diffuse through the cell membrane. Its existence inside the cells is generally due to the reduction of Cr VI, the endocytosis, or the absortion by the cells via phagocytosis. Cr III acts directly on the DNA molecule, while Cr VI reacts little with this molecule. In the ecosystem, however, Cr VI is more dangerous since this is the form that presents greater reactivity with biological membranes, crossing them and being easily incorporated into the cell. In the cell it is biotransformed to Cr III, a potentially mutagenic molecule. In vivo and in vitro studies have shown that organisms exposed to Cr VI present greater induction to a variety of damages to the DNA molecule. Among the damages induced by Cr, changes in the structure of the DNA molecule have been reported, with breaks of the major chain and base oxidation. In the organisms, these alterations generate chromosomal aberrations, micronucleus formation, sister chromatid exchanges, and errors in DNA synthesis.

Physical-chemical and thermodynamic analyses of steam reforming of ethanol to hydrogen production

The steam reforming is one of most utilized process of hydrogen production because of its high production efficiencies and its technological maturity. The use of ethanol for this purpose is a interesting option because this is a renewable and less environmentally offensive fuel. The objective of this study is evaluate the physical-chemical, thermodynamic and environmental analyses of steam reforming of ethanol. whose objective is to produce 0.7 Nm3/h of hydrogen to be used by a PEMFC of l kW. In this physical-chemical analysis, a global reaction of ethanol was considered. That is, the superheated ethanol and steam, at high temperatures, react to produce hydrogen and carbon dioxide. Beyond it's the simplest form to study the steam reforming of ethanol to hydrogen production, it's the case where occurs the highest production of hydrogen (the product to be used by fuel cells) and carbon dioxide, to be eliminated. But this reaction isn't real and depends greatly on the thermodynamic conditions of reforming, technical features of reformer system and catalysts. Other products generally formed (but not investigated in this study) are methane, carbon monoxide, among others. It was observed that the products is commonly produced in the moment when the reaction attains temperatures about 206°C (below this temperature, the reaction trend to the reaetants, that is, from hydrogen and carbon dioxide to steam and ethanol) and the advance degree of this reaction increases when the temperature of reaction also increases and when its pressure decreases. It's suggested reactions at about 600°C or higher. However, when the temperature attains 700°C, the stability of this reaction is occurred, that is, the production of reaction productions attains to the limit, that is the highest possible production. In temperatures above 700°C, the use of energy is very high for produce more products, having higher costs of production that the suggested temperature. The indicated pressure is 1 atm., a value that allows a desirable economy of energy that would also be used for pressurization or depressurization of steam reformer. In exergetic analysis, it's seem that the lower irreversibililies occur when the pressure of reactions are lower. However, the temperature changes don't affect significantly the irreversibilites. Utilizing the obtained results from this analysis, it was concluded that the best thermodynamic conditions for steam reforming of ethanol is the same conditions suggested in the physical-chemical analysis. The exergetic and first law efficiencies are high on the thermodynamie conditions studied.

Genetic vaccine for tuberculosis (pVAXhsp65) primes neonate mice for a strong immune response at the adult stage

Background: Vaccination of neonates is generally difficult due to the immaturity of the immune system and consequent higher susceptibility to tolerance induction. Genetic immunization has been described as an alternative to trigger a stronger immune response in neonates, including significant Th1 polarization. In this investigation we analysed the potential use of a genetic vaccine containing the heat shock protein (hsp65) from Mycobacterium leprae (pVAXhsp65) against tuberculosis (TB) in neonate mice. Aspects as antigen production, genomic integration and immunogenicity were evaluated. Methods: Hsp65 message and genomic integration were evaluated by RT-PCR and Southern blot, respectively. Immunogenicity of pVAXhsp65 alone or combined with BCG was analysed by specific induction of antibodies and cytokines, both quantified by ELISA. Results: This DNA vaccine was transcribed by muscular cells of neonate mice without integration into the cellular genome. Even though this vaccine was not strongly immunogenic when entirely administered (three doses) during early animal's life, it was not tolerogenic. In addition, pVAXhsp65 and BCG were equally able to prime newborn mice for a strong and mixed immune response (Th1 + Th2) to pVAXhsp65 boosters administered later, at the adult life. Conclusion: These results suggest that pVAXhsp65 can be safely used as a priming stimulus in neonate animals in prime-boost similar strategies to control TB. However, priming with BCG or pVAXhsp65, directed the ensuing immune response triggered by an heterologous or homologous booster, to a mixed Th1/Th2 pattern of response. Measures as introduction of IL-12 or GM-CSF genes in the vaccine construct or even IL-4 neutralization, are probably required to increase the priming towards Th1 polarization to ensure control of tuberculosis infection. © 2007 Pelizon et al; licensee BioMed Central Ltd.

Diferenciação hepatocítica de células-tronco mesenquimais da medula óssea humana

Some recent articles have reported that mesenchymal stem cells (MSCs) can be induced to express hepatocyte markers by transplanting them into animal models of liver damage, or by in vitro culture with growth factors and cytokines. In this study, the aim is to evaluate the behavior of MSCs subjected to induction of hepatocyte differentiation. The MSCs were isolated from the bone marrow of 4 normal donors, characterized and subjected to both in vitro and in vivo induction of hepatocyte differentiation. The in vitro induced cells showed morphological changes, acquiring hepatocyte-like features. However, the immunophenotype of these cells was not modified. The induced cells exhibited no increase in albumin, cytokeratin 18 or cytokeratin 19 transcripts, when analyzed by real-time RT-PCR. The expression of albumin, cytokeratin 18 and alpha fetoprotein was also unchanged, according to immunofluorescence tests. In vivo, the MSC demonstrated a potential to migrate to damaged liver tissue in immunodeficient mice. Taken together, the results suggest that bone marrow MSCs are incapable of in vitro differentiation into hepatocytes by the approach used here, but are capable of homing to damaged hepatic tissue in vivo, suggesting a role for them in the repair of the liver. This contribution to tissue repair could be associated with a paracrine effect exerted by these cells.

The importance of hormone receptor analysis in osteosarcoma cells growth submitted to treatment with estrogen in association with thyroid hormone

Bone tumor incidence in women peaks at age 50-60, coinciding with the menopause. That estrogen (E2) and triiodothyronine (T3) interact in bone metabolism has been well established. However, few data on the action of these hormones are available. Our purpose was to determine the role of E2 and T3 in the expression of bone activity markers, namely alkaline phosphatase (AP) and receptor activator of nuclear factor κB ligand (RANKL). Two osteosarcoma cell lines: MG-63 (which has both estrogen (ER) and thyroid hormone (TR) receptors) and SaOs-29 (ER receptors only) were treated with infraphysiological E2 associated with T3 at infraphysiological, physiological, and supraphysiological concentrations. Real-time RT-PCR was used for expression analysis. Our results show that, in MG-63 cells, infraphysiological E2 associated with supraphysiological T3 increases AP expression and decreases RANKL expression, while infraphysiological E2 associated with either physiological or supraphysiological T3 decreases both AP and RANKL expression. On the other hand, in SaOs-2 cells, the same hormone combinations had no significant effect on the markers' expression. Thus, the analysis of hormone receptors was shown to be crucial for the assessment of tumor potential growth in the face of hormonal changes. Special care should be provided to patients with T3 and E2 hormone receptors that may increase tumor growth. Copyright © 2007 John Wiley & Sons, Ltd.

Effect of 6-months of physical exercise on the nitrate/nitrite levels in hypertensive postmenopausal women

Background: Evidences have showed that the incidence of arterial hypertension is greater in postmenopausal women as compared to premenopausal. Physical inactivity has been implicated as a major contributor to weight gain and abdominal obesity in postmenopausal women and the incidence of cardiovascular disease increases dramatically after menopause. Additionally, more women than men die each year of coronary heart disease and are twice as likely as men to die within the first year after a heart attack. A healthy lifestyle has been strongly associated with the regular physical activity and evidences have shown that physically active subjects have more longevity with reduction of morbidity and mortality. Nitric oxide (NO) produced by endothelial cells has been implicated in this beneficial effect with improvement of vascular relaxing and reduction in blood pressure in both laboratory animals and human. Although the effect of exercise training in the human cardiovascular system has been largely studied, the majority of these studies were predominantly conducted in men or young volunteers. Therefore, the aim of this work was to investigate the effects of 6 months of dynamic exercise training (ET) on blood pressure and plasma nitrate/nitrite concentration (NOx-) in hypertensive postmenopausal women. Methods: Eleven volunteers were submitted to the ET consisting in 3 days a week, each session of 60 minutes during 6 months at moderate intensity (50% of heart rate reserve). Anthropometric parameters, blood pressure, NOx- concentration were measured at initial time and after ET. Results: A significant reduction in both systolic and diastolic blood pressure values was seen after ET which was accompanied by markedly increase of NOx- levels (basal: 10 ± 0.9; ET: 16 ± 2 μM). Total cholesterol was significantly reduced (basal: 220 ± 38 and ET: 178 ± 22 mg/dl), whereas triglycerides levels were not modified after ET (basal: 141 ± 89 and ET: 147 ± 8 mg/dl). Conclusion: Our study shows that changing in lifestyle promotes reducftion of arterial pressure which was accompanied by increase in nitrite/nitrate concentration. Therefore, 6-months of exercise training are an important approach in management arterial hypertension and play a protective effect in postmenopausal women. © 2009 Zaros et al; licensee BioMed Central Ltd.

Evidence for 2.35 to 2.30 Ga juvenile crustal growth in the northwest Borborema Province, NE Brazil

The c. 600 Ma Brasiliano Borborema Province of NE Brazil comprises a complex collage of Precambrian crustal blocks cut by a series of continental-scale shear zones. The predominant basement rocks in the province are 2.1-2.0 Ga Transamazonian gneisses of both juvenile and reworked nature. U-Pb zircon and Sm-Nd whole-rock studies of tonalite-trondhjemite-granodiorite basement gneisses in the NW Ceará or Médio Coreaú domain in the northwestern part of the Borborema Province indicate that this represents a continental fragment formed by 2.35-2.30 Ga juvenile crust. This block has no apparent genetic affinity with any other basement gneisses in the Borborema Province, and it does not represent the tectonized margin of the c. 2.1-2.0 Ga São Luis Craton to the NW. The petrological and geochemical characteristics, as well as the Nd-isotopic signatures of these gneisses, are consistent with their genesis in an island arc setting. This finding documents a period of crustal growth during a period of the Earth's history which is known for its tectonic quiescence and paucity of crust formation. © Geological Society of London 2009.

Antitumor properties of Ganoderma lucidum polysaccharides and terpenoids

Ganoderma lucidum is an edible medicinal mushroom with immunomodulatory and antitumor properties, which are mainly attributed to polysaccharides and triterpenes that can be isolated from mycelia, fruiting bodies and spores. G. lucidum has been us d in a powdered form, as a medicinal beverage and a nutraceutical food (usually dried). In the present review we report some historical facts and the experimental evidence that polysaccharides and triterpenes obtained from this mushroom present potential antitumor activity. Direct effects on tumor cells include induction of apoptosis and interference in the cell cycle, whereas indirect effects are based on the modulation of immune response, usually impaired by cancer cells. Data indicate that G. lucidum can be used as a complementary tool for treatment of cancer patients. © by São Paulo State University.

Simple method for culture of peripheral blood lymphocytes of Testudinidae.

We developed and optimized a simple, efficient and inexpensive method for in vitro culture of peripheral blood lymphocytes from the Brazilian tortoise Chelonoidis carbonaria (Testudinidae), testing various parameters, including culture medium, mitogen concentration, mitotic index, culture volume, incubation time, and mitotic arrest. Peripheral blood samples were obtained from the costal vein of four couples. The conditions that gave a good mitotic index were lymphocytes cultured at 37°C in minimum essential medium (7.5 mL), with phytohemagglutinin as a mitogen (0.375 mL), plus streptomycin/penicillin (0.1 mL), and an incubation period of 72 h. Mitotic arrest was induced by 2-h exposure to colchicine (0.1 mL), 70 h after establishing the culture. After mitotic arrest, the cells were hypotonized with 0.075 M KCl for 2 h and fixed with methanol/acetic acid (3:1). The non-banded mitotic chromosomes were visualized by Giemsa staining. The diploid chromosome number of C. carbonaria was found to be 52 in females and males, and sex chromosomes were not observed. We were able to culture peripheral blood lymphocytes of a Brazilian tortoise in vitro, for the preparation of mitotic chromosomes.