Characterization of a monoclonal antibody recognizing mast cells

Immunohistochemical screening for monoclonal antibodies prepared by immunization of mice with a rat osteoblastic cell population led to identification of one antibody that reacted against a small population of cells present in the soft connective tissue compartment of 21 days fetal rat calvaria. The morphology of the cells and the immunohistochemical staining characteristics (a distinct intracellular granular pattern) suggested that the antibody might be reacting specifically against mast cells. We used combined histochemistry and immunohistochemistry to further characterize this antibody, designated RCJ102. Cryosections containing calvaria bone, soft connective tissues and skin were prepared from the top of the head of 21 days fetal rats, and from adult rats cryosections of lung, muscle, adipose tissue and small intestine were prepared. Some sections were labelled by indirect immunofluorescence with RCJ102; corresponding sections were labelled histochemically with toluidine blue. There was a direct correspondence between mast cells identified histochemically and cells labelling with RCJ102 in all tissues except intestine, in which the mast cell detectable by histochemistry were not labelled by RCJ102. These results suggest that the RCJ102 antibody will be a valuable new reagent for further elucidation of the heterogeneity described between connective tissue and intestinal mucosal mast cells.

VapB type 8 plasmids in Rhodococcus equi isolated from the small intestine of pigs and comparison of selective culture media

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Experimental glaucoma model (ischemia and reperfusion): histology, morphometry, protein and gene espression of apoptosis pathway

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

Hyaluronidase splice variants are associated with histology and outcome in adenocarcinoma and squamous cell carcinoma of the lung

Heterogeneity of hyaluronidase (HYAL) expression has been identified in tumors and shows promise as an indicator of disease progression. The expression profile of alternatively spliced forms of HYAL was evaluated in tumors and normal lung tissue from 69 resected tumors of patients with adenocarcinomas and squamous cell carcinomas. HYAL1-wild-type (wt) and variants 1 to 5, HYAL2-wt, and HYAL3-wt, and variants 1 to 3 were identified by polymerase chain reaction and direct sequencing. Different proportions of the 3 HYAL-wt and variants were expressed in tumor and normal lung tissues. HYAL1-wt was associated with a poorer prognosis and HYAL3-vl with a better prognosis. HYAL splice variants are associated with histology and outcome, suggesting that strategies aimed at modulating their levels may be effective for lung cancer treatment. (C) 2012 Elsevier Inc. All rights reserved.

Intestinal IgG uptake by small intestine of goat kid fed goat or lyophilized bovine colostrum

The immunoglobulin G (IgG) uptake and enterocyte nucleus position in the villous were studied in newborn goat kids fed goat or lyophilized bovine colostrum. Two groups of 15 newborn goat kids, each received 5% of body weight of goat colostrum (GC) or lyophilized bovine colostrum (LBC) containing 55 mg/mL of immunoglobulin G (IgG) at 0, 7 and 14 h of life. Three animals were sampled just after birth, receiving no colostrum intake, to be used as control. Samples of duodenum, medium jejunum and ileum were collected at 0, 18, 36 and 96 h of life. IgG vacuoles were not observed in the duodenum throughout the experiment regardless of all the experimental time points. In this segment, at 0, 18 and 36 h of life, nuclei were found in the apical, medial and basal positions in the enterocytes, and localized in the upper, medial and lower parts in the villous, respectively. At 96 h, a basal nuclei position was observed in the enterocytes, throughout the villous. In jejunum, IgG vacuoles were distributed along the villous at 18 and 36 h. In this segment at Oh the nuclei were positioned predominantly apically in the enterocytes, throughout the villous. At 18 and 36 h, no consistent nuclei pattern was verified: however at 96 h, the nuclei were positioned basally in the enterocytes, throughout the jejunal villous. In the ileum at 0, 18 and 36 h, a great number of vacuoles without IgG were verified in the medial-apical part of the villous. In this segment, at Oh of life and 96 h of life, the predominance of basal nuclei was observed. Nuclei were positioned in medial-apically part of the ileal enterocytes in the upper part of the villous at 18 and 36 h. It was found that the jejunal epithelium was the most important segment related to absorption process. The IgG absorption and nucleus position in the newborn goats were dependent on the small intestine segments and experimental time points, regardless of the colostrum source. GC or LCB. Considering the IgG uptake mechanism observed in the present study, the lyophilized bovine colostrum might be used instead of goat colostrum. (C) 2011 Elsevier B.V. All rights reserved.

Histology, histochemistry and stereology of the adipose fin of Prochilodus lineatus

The adipose fin is small, nonpared, and usually located medially between the dorsal and caudal fin. Its taxonomic occurrence is very restrict; thus, it represents an important trace for taxon distinction. As it does not play a known vital physiological roll and it is easily removed, it is commonly used in marking and recapture studies. The present study characterizes the adipose fin of Prochilodus lineatus, as it is poorly explored by the literature. The adipose fin consists basically of a loose connective core, covered by a stratified epithelium supported by collagen fibers. At the epithelium, pigmented cells and alarm substance cells are found. Despite the name, adipocytes or lipid droplets are not observed on the structure of the fin. Microsc. Res. Tech., 2012. (c) 2011 Wiley Periodicals, Inc.

Enzyme activity in the small intestine of goat kids during the period of passive immunity acquisition

Enzyme activity of protein and carbohydrate degradation in small intestinal mucosa was investigated in goat kids fed with lyophilized bovine and goat colostrum. At 0,7 and 14 h of life 15 male newborns received 5% of body weight of lyophilized bovine colostrum and 14 goat colostrum, both with 55 mg/mL of IgG. Duodenum, jejunum and ileum samples were collected at 18,36 and 96 h of life. Three animals were sampled at birth, without colostrum intake. Activity of aminopeptidase N and A, dipeptidil peptidase IV, lactase, maltase and sucrase was determined as one international unit per gram of tissue. Intracellular enzymatic activity of acid phosphatase was observed by histochemistry in tissue section. Only the activity of aminopeptidase A in the ileum was affected by treatment, with a greater value for LBC than for GC (P < 0.05). The aminopeptidase N activity was the highest at 36 h in the duodenum (P < 0.05) and lowest at 96 h in the jejunum (P < 0.05). Dipeptidil peptidase IV activity was highest at 36 h in the duodenum (P < 0.05), lowest at 96 h in the jejunum (P < 0.05) and higher at 36 h than at 96 h in the ileum (P < 0.05). Aminopeptidase A activity in the ileum was highest at 36 h (P < 0.05), followed by 18 and 96 h of life (P < 0.05). Lactase activity in the duodenum increased from 18 to 36 h and from 36 to 96 h in the jejunum (P < 0.05). Maltase activity increased only in the duodenum from 18 to 96 h (P < 0.05). Sucrase activity in the jejunum decreased from 18 to 36 h and from 36 to 96 h in the ileum (P < 0.05). At birth, activity of most enzymes was similar to that at later times (P < 0.05). Histochemistry analyses showed a higher frequency of lysosomes with acid phosphatase activity in the duodenum, especially at 36 h of life. In the jejunum, the presence of lysosomes with acid phosphatase activity was the highest at 96 h, followed by 36 and 18 h of life. In the ileum, all samples showed low presence of lysosomes with acid phosphatase activity. These results indicate that lyophilized bovine colostrum, as a heterologous source of antibodies or nutrients, is a possible alternative management tool for goats. The present work also suggests that in the first 4 days of life, enzyme activity in the intestinal epithelium of goats is still not fully stimulated, which is an important characteristic for these animals that depend on macromolecule absorption to acquire passive protection after birth. (C) 2012 Elsevier B.V. All rights reserved.

Regular and moderate exercise before experimental sepsis reduces the risk of lung and distal organ injury

de Araujo CC, Silva JD, Samary CS, Guimaraes IH, Marques PS, Oliveira GP, do Carmo LGRR, Goldenberg RC, Bakker-Abreu I, Diaz BL, Rocha NN, Capelozzi VL, Pelosi P, Rocco PRM. Regular and moderate exercise before experimental sepsis reduces the risk of lung and distal organ injury. J Appl Physiol 112: 1206-1214, 2012. First published January 19, 2012; doi:10.1152/japplphysiol.01061.2011.-Physical activity modulates inflammation and immune response in both normal and pathologic conditions. We investigated whether regular and moderate exercise before the induction of experimental sepsis reduces the risk of lung and distal organ injury and survival. One hundred twenty-four BALB/c mice were randomly assigned to two groups: sedentary (S) and trained (T). Animals in T group ran on a motorized treadmill, at moderate intensity, 5% grade, 30 min/day, 3 times a week for 8 wk. Cardiac adaptation to exercise was evaluated using echocardiography. Systolic volume and left ventricular mass were increased in T compared with S group. Both T and S groups were further randomized either to sepsis induced by cecal ligation and puncture surgery (CLP) or sham operation (control). After 24 h, lung mechanics and histology, the degree of cell apoptosis in lung, heart, kidney, liver, and small intestine villi, and interleukin (IL)-6, KC (IL-8 murine functional homolog), IL-1 beta, IL-10, and number of cells in bronchoalveolar lavage (BALF) and peritoneal lavage (PLF) fluids as well as plasma were measured. In CLP, T compared with S groups showed: 1) improvement in survival; 2) reduced lung static elastance, alveolar collapse, collagen and elastic fiber content, number of neutrophils in BALF, PLF, and plasma, as well as lung and distal organ cell apoptosis; and 3) increased IL-10 in BALF and plasma, with reduced IL-6, KC, and IL-1 beta in PLF. In conclusion, regular and moderate exercise before the induction of sepsis reduced the risk of lung and distal organ damage, thus increasing survival.

Heme oxygenase-1 activity is involved in the control of Toxoplasma gondii infection in the lung of BALB/c and C57BL/6 and in the small intestine of C57BL/6 mice

Heme oxygenase-1 (HO-1) is an enzyme that catabolizes free heme, which induces an intense inflammatory response. The expression of HO-1 is induced by different stimuli, triggering an anti-inflammatory response during biological stress. It was previously verified that HO-1 is able to induce indoleamine 2,3-dioxygenase (IDO), an enzyme that is induced by IFN-γ in Toxoplasma gondii infection. To verify the role of HO-1 during in vivo T. gondii infection, BALB/c and C57BL/6 mice were infected with the ME49 strain and treated with zinc protoporphyrin IX (ZnPPIX) or hemin, which inhibit or induce HO-1 activity, respectively. The results show that T. gondii infection induced high levels of HO-1 expression in the lung of BALB/c and C57BL6 mice. The animals treated with ZnPPIX presented higher parasitism in the lungs of both lineages of mice, whereas hemin treatment decreased the parasite replication in this organ and in the small intestine of infected C57BL/6 mice. Furthermore, C57BL/6 mice infected with T. gondii and treated with hemin showed higher levels of IDO expression in the lungs and small intestine than uninfected mice. In conclusion, our data suggest that HO-1 activity is involved in the control of T. gondii in the lungs of both mouse lineages, whereas the hemin, a HO-1 inducer, seems to be involved in the control of parasitism in the small intestine of C57BL/6 mice.

Comparative study by computed radiography, histology, and scanning electron microscopy of the articular cartilage of normal goats and in chronic infection with caprine arthritis-encephalitis virus

In the northeast of Brazil, caprine arthritis-encephalitis (CAE) is one of the key reasons for herd productivity decreasing that result in considerable economic losses. A comparative study was carried out using computed radiography (CR), histological analysis (HA), and scanning electronic microscopy (SEM) of the joints of CAE infected and normal goats. Humerus head surface of positive animals presented reduced joint space, increased bone density, and signs of degenerative joint disease (DJD). The carpal joint presented no morphological alterations in CR in any of the animals studied. Tarsus joint was the most affected, characterized by severe DJD, absence of joint space, increased periarticular soft tissue density, edema, and bone sclerosis. Histological analysis showed chronic tissue lesions, complete loss of the surface zone, absence of proteoglycans in the transition and radial zones and destruction of the cartilage surface in the CAE positive animals. Analysis by SEM showed ulcerated lesions with irregular and folded patterns on the joint surface that distinguished the limits between areas of normal and affected cartilage. The morphological study of the joints of normal and CAE positive goats deepened understanding of the alteration in the tissue bioarchitecture of the most affected joints. The SEM finding sustained previous histological reports, similar to those found for rheumatoid arthritis, suggesting that the goat infected with CAE can be considered as a potential model for research in this area.

Effect of different live prey, Grapsus grapsus Linnaeus 1758 and Plagusia depressa Fabricius, 1775 zoeas,over histology and biochemical composition of common octopus, octopus vulgaris Cuvier, 1797 paralarvae

[EN] Since paralarval rearing is still the main bottleneck for the development of octopus culture, the aim of the present study was to obtain some information on the feeding strategy and nutritional requirements during paralarval stage. For that purpose just hatched out octopus paralarvae were fed with live preys in three different combinations, trying to match their natural food: Enriched Artemia metanauplii, Grapsus grapsus zoeas supplemented with enriched Artemia, and Plagusia depressa zoeas supplemented with enriched Artemia. Paralarval treatments were carried out during 28 days in triplicates; fibre glass 120 l tanks in flow through system were used. Growth, in terms of dry body weight, mantle length and width, was determined each seven days. A histological study of the paralarval development was carried out. Biochemical composition of preys and paralarvae were determined. Growth was significantly better in paralarvae fed with zoeas and Artemia than in those fed only with Artemia, from day 8 after hatching. Besides a clear effect on the digestive gland histology morphology was observed.

The effects of supplementation of decapod zoea to Artemia basal diet on the digestive gland histology and the fatty acid composition in common octopus (Octopus vulgaris) paralarvae.

[EN]The present study aimed to evaluate the effect of the supplementation of different crab zoeas to enriched Artemia basal diet for O. vulgaris paralarvae during the first month of life. Paralarvae were fed using: enriched Artemia nauplii alone and Artemia co-fed with either first zoea stages of Grapsus adscensionis or Plagusia depressa. The experiment was carried out over a period of 28 days, in 0.12 m3 tanks with a flow-through rearing system. Growth in dry weightas well as mantle length and width were assessed weekly. Additionally, prey and paralarvae fatty acid composition and digestive gland (DG) histology were evaluated.

A new histology scoring system for the assessment of the quality of human cartilage repair: ICRS II

A reliable and reproducible method is needed to assess cartilage repair.

Plaque characteristics of nonobstructive coronary lesions in diabetic patients: an intravascular ultrasound virtual histology analysis

Patients with diabetes mellitus are known to be at increased risk for acute cardiovascular events. We used intravascular ultrasound virtual histology (IVUS-VH) to examine whether nonobstructive coronary artery lesions of diabetic patients have distinct plaque composition and morphology compared with nondiabetic patients.

Analysis of coronary bifurcations by intravascular ultrasound and virtual histology

Background Regional differences in shear stress have been identified as reason for early plaque formation in vessel bifurcations. We aimed to investigate regional plaque morphology and composition using intravascular ultrasound (IVUS) and virtual histology (IVUS–VH) in coronary artery bifurcations. Methods We performed IVUS and IVUS–VH studies at coronary bifurcations to analyze segmental plaque burden and composition of different segments in relation to their orientation to the bifurcation. Results A total of 236 patients with a mean age of 59 ± 11 years (69% male) were analyzed. Plaque burden was higher at the contralateral vessel wall facing the bifurcation compared to the ipsilateral vessel wall and this difference was true for proximal and distal segments (proximal: 37 ± 12% and 45 ± 15% for segments at the ipsilateral and contralateral vessel wall, respectively, p < 0.001; distal: 37 ± 10% and 47 ± 15% for segments at the ipsilateral and contralateral vessel wall, respectively, p < 0.001). In addition, these segments exhibited a higher proportion of dense calcium and a lower proportion of fibrous tissue and fibro fatty tissue. Conclusions Segments on the contralateral wall of the bifurcation which have previously been identified as regions with low shear stress not only exhibited a higher plaque burden, but also a higher degree of calcification.