Infectious bronchitis virus replication in the chicken embryo related cell line

The susceptibility of the chicken embryo related (CER) cell line to infectious bronchitis virus (IBV M41) was characterized after five consecutive passages in CER cells. Virus replication was monitored by cytopathic effect observation, electron microscopy, indirect immunofluorescence, and reverse transcription polymerase chain reaction (RT-PCR). At 96 h post-infection (p.i.), the cytopathic effect was graded 75% by cell fusion, rounding up of cells and monolayer detachment, and the electron microscopy image characterized by coronavirus morphology. Cytoplasmic fluorescence was readily observed by from 24 h p.i. onwards, and at all times the respective viral RNA from IBV-infected monolayers was demonstrated by RT-PCR. Extra-cellular virus was measured by virus titration performed on chicken kidney cells and embryonated chicken eggs, and respective titres ranged from 4.0 to 6.0 log(10) EID50/ml on embryonated chicken eggs, and from 2.0 to 6.0 log(10) TCID50/ml on both CER cells and chicken kidney cells studied from 24 to 120 h p.i. These results confirmed that the M41 strain replicated well in the CER cell line.

Susceptibility of mammalian cell line for isolation of IBDV from clinical samples

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Transportadores de glicose na síndrome metabólica

A regulação da homeostasia intra e extra-celular da glicose está diretamente relacionada ao controle preciso da expressão dos genes que codificam as diferentes isoformas de proteínas transportadoras de glicose, as quais se expressam de maneira tecido-específica, em conseqüência do padrão de ativação dos fatores transcricionais reguladores de cada gene, em cada tipo celular. A síndrome metabólica (SM) abrange uma grande variedade de alterações fisiopatológicas, todas de repercussões sistêmicas, acometendo os mais distintos territórios do organismo, nos quais alterações nos transportadores de glicose presentes são observadas em maior ou menor grau. A presente revisão abordará as alterações na expressão de transportadores de glicose claramente demonstradas na literatura, cujas repercussões nos fluxos territoriais de glicose auxiliam na compreensão de mecanismos fisiopatológicos da SM, assim como dos tratamentos propostos para esta entidade.

Variation of Laser Intensity on the Healing of Cutaneous Wounds

Tissue repair is an integration of dynamic interactive processes that involves soluble mediators, blood components, production of extra-cellular matrix and mesenchymal cells. Many studies involving the use of LLLT shows that the healing process is favored by such therapy. The aim of this work was to evaluate, through histological analysis, the tissue effects of cutaneous wounds submitted to different intensities and a same irradiation dose with lasers in λ670 or λ685nm. Eighteen animals were divided in two experimental groups according to wavelength used (λ670 or λ685nm). Each one of these groups was divided still in three subgroups of three animals each, related to the intensity of applied irradiation (2, 15 or 25mW). Twelve animals acted as untreated controls and were not irradiated. The irradiation was carried out during seven days. The animals were sacrificed eight days after surgery. The specimens were removed, kept in 4% formaldehyde for 24 hours, routinely prepared to wax, stained with H&E and analyzed under light microscopy. The histological characteristics observed, so much in the irradiated animals, as in the control, they are indicative of a substitution repair process, however, the LLLT modulatory positive effect was observed, in the healing process, mainly associate to the use of the shorter wavelength and low power. The results of the present study indicate that LLLT improves cutaneous wound repair and best results are achieved when higher potencies associated to short wavelengths or lower potencies associated to higher wavelengths are used.

A preliminary report on the effect of laser therapy on the healing of cutaneous surgical wounds as a consequence of an inversely proportional relationship between wavelength and intensity: Histological study in rats

Objective: The objective of the present investigation was to assess the histological effects of different wavelengths and intensities on the healing process of cutaneous wounds. Background Data: Tissue repair is a dynamic interactive process which involves mediators, cells and extra-cellular matrix. Several reports on the use of laser therapy have shown that the healing process is positively affected when the correct parameters are used. Methods: Eighteen standardized wounds were surgically created on the dorsum of male and female Wistar rats, which were subsequently divided into two experimental groups according to wavelength used λ.670 or λ685 nm) for lasertherapy (LLLT). Each group was divided into three subgroups of three animals according to the intensity of the applied irradiation (2,15, or 25 mW). Twelve animals were used as entreated controls and were not irradiated. The irradiation was carried out during seven consecutive days. The animals were sacrificed eight days after surgery. The specimens were removed, kept in 4% formaldehyde for 24 h, routinely prepared to wax, stained with H&E, and analyzed under light microscopy. Results: For both groups, light microscopy showed a substitution repair process; however, when LLLT was used, a positive biomodulatory effect was detectable, chiefly associated with shorter wavelength and low intensity. Conclusions: The results of the present study indicate that LLLT improved cutaneous wound repair and that the effect is a result of an inversely proportional relationship between wavelength and intensity, with treatment more effective when combining higher intensity with short wavelength or lower intensity with higher wavelength.

Chronic renal failure in male and female rats

Gender influences the progression of chronic renal failure (CRF). We studied male (M) and female (F) Wistar rats for 90 days: castrated (CMc, n=7; CFc, n=6) and non castrated controls (CM, n=9; CF, n=6); castrated (CRFMc, n=8; CRFFc, n=6) and non castrated animals submitted to 5/6 nephrectomy (CRFM, n=13; CRFF, n=6). Data are expressed as mean ± SEM. Proteinuria (PTN) was higher in CRFM (554 ± 69mg/24h) compared to CRFMc (277 ± 85 mg/24h), but not in females (CRFF=193 ± 20mg/24h, CRFFc=164 ± 71mg/24h). Mesangial fractional volume increased in all CRF animals. CRF animals showed an increase of glomerular sclerosis index (GSI) and tubulointerstitial damage (TID) but in a smaller proportion in male castrated animals; the opposite occurred with females: castration induced an increase of these parameters. CRF animals showed increased cortical and glomerular fibronectin (FN) rates. Castration decreased glomerular and cortical FN rates in CRFM but not in females. In conclusion, proteinuria was higher in CRFM and probably led to glomerular and interstitial damage, as well as to FN accumulation, castration seems to protect against development of PTN, TID and FN accumulation in males. Castrated female rats presented mesangial expansion, with no changes in PTN, TID and FN rates. It seems that female sex hormones do not protect against renal disease progression, instead, we suggest that male sex hormones lead to acceleration of CRF.

Application of a serum/protein-free medium for the growth of mammalian cell lines and high level production of classical, variant and very virulent strain of infectious bursal disease virus (IBDV)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Matrix metalloproteinase gene polymorphisms and oral cancer

Since oral squamous cell carcinoma (OSCC) is the most prevalent malignant cancer in the oral cavity, several researches have been performed to study the role of important enzymes in this disease. Among them, the matrix metalloproteinases (MMPs) are highlighted, due to the fact that they are proteinases responsible to degrade many extra-cellular matrix components, making possible the invasion of neoplasic cells. Important tools in cancer prognosis have been utilized aiming to correlate high levels of MMPs and OSCC, such as immunohistochemical, zymographic and mRNA detection methods. However, these techniques are usually applied after cancer detection, characterizing a curative but not a preventive medicine. Trying to make interventions before the development of the disease and making possible the identification of people at high risk and, analysis of modifications in MMP genes has been a chance for modern medicine. Recently, polymorphisms in MMP genes have been related to different neoplasias, including OSCC. Despite investigation is beginning, MMP gene polymorphisms seems to have a promising future in oral cancer research and some of the present results have shown that there are MMP polymorphisms related to an increased risk for developing oral cancer. Key words:Oral cancer, polymorphism, matrix metalloproteinase.

Ação extra-nuclear do hormônio triiodotironina (T3) na expressão gênica de HIF-1α e TGFα em linhagem celular de adenocarcinoma mamário

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Calculable lepton masses, seesaw relations, and four neutrino mixings in a 3-3-1 model with an extra U(1) symmetry

We propose a scheme in which the masses of the heavier leptons obey seesaw type relations. The light lepton masses, except the electron and the electron neutrino ones, are generated by one loop level radiative corrections. We work in a version of the 3-3-1 electroweak model that predicts singlets (charged and neutral) of heavy leptons beyond the known ones. An extra U(1)(Omega) symmetry is introduced in order to avoid the light leptons getting masses at the tree level. The electron mass induces an explicit symmetry breaking at U(1)(Omega). We discuss also the mixing matrix among four neutrinos. The new energy scale required is not higher than a few TeV.

Intra- and extracellular osmotic regulation in the hololimnetic Caridea and Anomura: a phylogenetic perspective on the conquest of fresh water by the decapod Crustacea

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Kinetics of cellular component in inflammatory response induced by different stimuli in the swim bladder of pacu, Piaractus mesopotamicus Holmberg 1887 (Characidae)

The present study assessed the kinetics of cell accumulation at the site of inflammation induced by thioglycolate, Escherichia coli lipopolysaccharide (LPS) and heat-inactivated Aeromonas hydrophila, in the pacu, Piaractus mesopotamicus (Characidae), swim bladder. A quantitative, as well as qualitative, assessment was done of all the cells present in the exudate at 6, 24, and 48 h (n = 8) after inoculation of inflammatory agents. The results show that the thioglycolate was the irritant to induce higher total inflammatory cell accumulation when compared to the control group, 6 h after insult (P < 0.05). Inoculation of heat-inactivated Aeromonas hydrophila induced progressive accumulation of total inflammatory cells, with cell number peaking after 24 h and being significantly higher than observed in the other groups (P < 0.05). Injection of LPS also induced greater cell accumulation when compared to the control group (P < 0.05), although in lower numbers than those induced by the other two irritants. All irritants injected induced significantly greater accumulation of lymphocytes and thrombocytes when compared to the control group (P < 0.05).

Humoral and cellular immune response generated by different vaccine programs before and after Salmonella Enteritidis challenge in chickens

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cellular and molecular characterization of an embryonic cell line (BME26) from the tick Rhipicephalus (Boophilus) microplus

The cellular and molecular characteristics of a cell line (BME26) derived from embryos of the cattle tick Rhipicephalus (Boophilus) microplus were studied. The cells contained glycogen inclusions, numerous mitochondria, and vesicles with heterogeneous electron densities dispersed throughout the cytoplasm. Vesicles contained lipids and sequestered palladium meso-porphyrin (Pd-mP) and rhodamine-hemoglobin, suggesting their involvement in the autophagic and endocytic pathways. The cells phagocytosed yeast and expressed genes encoding the antimicrobial peptides (microplusin and defensin). A cDNA library was made and 898 unique mRNA sequences were obtained. Among them, 556 sequences were not significantly similar to any sequence found in public databases. Annotation using Gene Ontology revealed transcripts related to several different functional classes. We identified transcripts involved in immune response such as ferritin, serine proteases, protease inhibitors,. antimicrobial peptides, heat shock protein, glutathione S-transferase, peroxidase, and NADPH oxidase. BME26 cells transfected with a plasmid carrying a red fluorescent protein reporter gene (DsRed2) transiently expressed DsRed2 for up to 5 weeks. We conclude that BME26 can be used to experimentally analyze diverse biological processes that occur in R. (B.) microplus such as the innate immune response to tick-borne pathogens. (C) 2008 Elsevier Ltd. All rights reserved.