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Evaluate the effect of 12 wks of concurrent training (CT) in the extracellular matrix (ECM) of subcutaneous adipose tissue (SAT) in people living with HIV/AIDS (PLWHA). In the non-randomized clinical trial, 19 participants, 11 healthy (HIV–) and 18 PLWHA under the use of highly active antiretroviral therapy (HAART) for at least 1 year (HIV+). All participants engaged in a moderate-intensity CT program for 12 weeks, 3 times a week. Before and after CT, aerobic and strength performance were assessed, as well as anthropometric and biochemical blood profiles. In addition, SAT biopsies were performed for histologic and morphometric analyses. Statistical analysis was carried out with R Studio, using descriptive and inferential analysis, ANOVA test, and mixed-effect model (P < 0.05). HIV+ showed higher levels of very-low-density lipoproteins and triglycerides and lower levels of high-density lipoproteins at baseline than HIV– (P < 0.05). All groups showed improved aerobic and strength performances (P < 0.05). Both groups showed reduced adipocyte sizes after CT (P < 0.05). Lastly, HIV+ presented smaller adipocytes and higher elastic fiber deposition at baseline and decreased after training only in HIV+, similar to the HIV group. Thus, CT in PLWHA promoted a decrease in the size heterogeneity of adipocytes and elastic fiber deposition, remodeling the ECM, and improving the SAT fibrosis profile. Brazilian Clinical Trials Registry (ensaiosclinicos.gov.br – UTN: U1111-1214-3022).Novelty: Adipose tissue fibrosis is improved by training in people living with HIV. Concurrent training remodels adipose tissue extracellular matrix.

Aerobic exercise training (AER) may promote several adaptations in white adipose tissue (WAT), including a phenotypic change known as browning. The present study aimed at assessing if resistance exercise training (RES) would be as efficient as AER in inducing a brown-like adipocyte reprogramming in WAT. Thirty Swiss male mice were randomly divided into 3 groups with 10 animals each: 1) sedentary (SED), 2) AER, and 3) RES. After the adaptation training, an incremental test was performed at the beginning of each week to adjust training load. Mice were submitted to 8 wk of AER or RES. After the experimental period, inguinal and retroperitoneal WAT (iWAT and rpWAT) and brown adipose tissue (BAT) were collected. The prescription of AER and RES was effective in increasing the performance of both groups. Also, RES presented a lower body weight than AER/SED. AER and RES reduced the area of iWAT and rpWAT adipocytes and the lipid area of BAT, induced an increase of vascular endothelial growth factor (VEGF) and cluster of differentiation 31 (CD31) and uncoupling protein 1 (UCP-1), and increased the expression of selective genes of brown and beige phenotype in adipocytes after 8 wk. In general, we demonstrated here that AER and RES training similarly induced the browning of iWAT and rpWAT.NEW & NOTEWORTHY Aerobic exercise training (AER) induces the browning of white adipose tissue, turning adipocytes multilocular, highly vascularized and expressing uncoupling protein 1 (UCP-1). The current study compared the efficiency of resistance to aerobic exercise training to promote a brown-like phenotype. Our results suggest that both types of training similarly induce subcutaneous and visceral adipose tissue browning.

Paracoccidioides brasiliensis (Pb) can disseminate through the lymphatic and hematogenic pathways. As a result the spleen and other lymphoid organs are targets of paracoccidioidomycosis. There are few studies describing this disease in the spleen and more detailed descriptions are required. This study combines Histopathology and Fourier Transform Raman Spectroscopy (FT-Raman) methods to study spleen infected by Pb. The Swiss mice were euthanized after 1, 2, 4 and 8 weeks of infection with Pb, and their spleens were removed for ex vivo analyzes. Histopathological evaluation revealed that the red pulp was the area most affected, presenting the highest concentration of yeasts, inflammatory cells and cells in apoptosis, with no observation of typical granuloma formation. The main physical-chemical changes were detected in the spectral ranges between 1730-1590 cm-1 and 1390-1280 cm-1 attributed to amide I and vibrational deformation mode of CH3 and CH2 molecules, respectively. The amide I was correlated to total protein content and CH3 and CH2 molecules to collagen of the spleen. The association between histological and physical-chemical methods enabled detection of several alterations in the spleen, including apoptosis, contributing to a better understanding of paracoccidioidomycosis, a tropical neglected disease.

This study evaluated whether anethole attenuates the inflammatory response and joint damage in a model of adjuvant-induced arthritis (AIA) in rats. The animals were treated with 62.5-, 125-, or 250-mg/kg anethole daily for 21 days after AIA and necropsied on days 14 and 21 to evaluate the number of serum and synovial leukocytes (total and differential), serum cytokines (IL-2, IL-6, IL-12, IL-17, and TNF-α), and nitric oxide concentrations. Morphologic changes in the cartilage and bone of the femorotibial articulation in both left paw and right paw were studied in hematoxylin/eosin and Sirius Red-hematoxylin sections. Different doses of anethole suppressed paw swelling and the number of serum and synovial leukocytes. However, 250 mg/kg of anethole more effectively controlled local and systemic inflammation. Histological evaluation revealed significant prevention of cartilage damage and inflammatory infiltrate scores. Morphometric analysis showed pannus formation, the thickness of the articular cartilage, and bone resorption lower in the anethole-treated AIA group compared to untreated AIA group on both days 14 and 21. These significant anti-inflammatory effects in the anethole-treated AIA group were associated with downregulation of cytokines and nitric oxide levels. Therefore, anethole may be a useful intervention to treat inflammatory arthritis.

In this study was investigated the effect of treatment with a fish oil preparation (FOP) on inflammatory and histopathological parameters in a rat model of adjuvant-induced arthritis (AIA). Paw edema, the severity of secondary lesions, the number of synovial leukocytes, and serum nitric oxide and cytokines concentrations were evaluated. Histological changes in cartilage and bone of the femorotibial articulation in the paws were analyzed by hematoxylin/eosin and sirius red-hematoxylin staining. Oral treatment with the FOP (75, 150, and 300 mg/kg) that was initiated on the day of arthritis induction (day zero) and maintained for 21 days inhibited inflammatory signals and prevented morphological changes in cartilage and bone of the femorotibial articulation of the paws. These effects could be at least partially attributable to inhibitory and modulatory actions of the FOP on the production and release of nitric oxide and cytokines that participate in the pathophysiology of AIA. En este estudio se investigó el efecto del tratamiento con una preparación de aceite de pescado (PAP) sobre los parámetros inflamatorios y histopatológicos en un modelo de artritis inducida por adyuvante en ratas (AIA). Se evaluó el edema de la pata, la gravedad de las lesiones secundarias, el número de leucocitos sinoviales y las concentraciones séricas de óxido nítrico y citocinas. Los cambios histológicos en el cartílago y el hueso de la articulación femorotibial en las patas se analizaron mediante tinción con hematoxilina / eosina y rojo de sirio-hematoxilina. El tratamiento oral con PAP (75, 150 y 300 mg / kg) que se inició el día de la inducción de la artritis (día zero) y se mantuvo durante 21 días inhibió las señales inflamatorios y previno cambios morfológicos en el cartílago y hueso de la articulación femorotibial de las patas. Estos efectos podrían atribuirse, al menos en parte, a las acciones inhibidoras y moduladoras de la PAP sobre la producción y liberación de óxido nítrico y citocinas que participan en la fisiopatología de la AIA. Nesse estudo foi investigado o efeito do tratamento com uma preparação de óleo de peixe (POP) sobre parâmetros inflamatórios e histopatológicos na artrite induzida por adjuvante em ratos (AIA). O edema de pata, a gravidade das lesões secundárias, o número de leucócitos sinoviais e a concentração sérica de óxido nítrico e de citocinas foram avaliados. Alterações histológicas nas cartilagens e nos ossos das articulações fêmoro-tibiais das patas foram analisadas pelas técnicas de coloração hematoxilina/eosina e sirius red-hematoxilina. O tratamento oral com a POP (75, 150, e 300 mg/kg) iniciado no dia da indução da artrite (dia zero) e mantido por 21 dias inibiu os sinais inflamatórios e preveniu as alterações morfológicas na cartilagem e no osso das articulações. Esses efeitos podem ser parcialmente atribuídos à ação modulatória e inibitória da POP sobre a produção/liberação de óxido nítrico e citocinas que participam da fisiopatologia da AIA.

The purpose of this study is to evaluate the toxicity of synthetic chalcones 1 and 2 in uninfected hamsters and anti-Leishmania activity of synthetic chalcones 1 and 2 in hamsters infected with Leishmania (Viannia) braziliensis. For the toxicity test, uninfected animals were treated with chalcones 1 and 2, and clinical and biochemical parameters and histological aspects of the liver and kidneys were assessed. Chalcones 1 and 2 were then intraperitoneally or topically administered (10 mg/kg body weight) three times per week in animals infected with promastigotes of L. (V.) braziliensis. We monitored the thickness of the infected footpads, determined parasitic load, performed histological analysis, and detected apoptosis in situ. The results were analyzed using Student’s t test and Mann-Whitney test at a significance level of 5 %. Neither of the chalcones showed toxicity. Chalcone 2 administered intraperitoneally significantly reduced the thickness of the infected footpad compared with the beginning of treatment. The parasite load of the lymph node and spleen was reduced in the groups treated with chalcones 1 (topical) and 2 (intraperitoneal). Chalcone 2 (topical) reduced parasite burden only in the lymph node. The histological analysis revealed reconstitution of the tissue and reductions of inflammation and apoptosis in the infected footpad in these groups. The synthetic chalcones 1 (topical) and 2 (intraperitoneal and topical) at a dose of 10 mg/kg showed anti-Leishmania activity in vivo, no renal or hepatic toxicity, and a reduction of apoptosis of the cells in the lesions. These chalcones may have substantial potential for the treatment of cutaneous leishmaniasis.

OBJECTIVE: To evaluate the effect of two different concentrations of sodium alendronate on the quantity of organic matrix in the femur of rats with estrogen suppression caused by ovariectomy.METHODS: Sixty-days-old Wistar rats (Rattus norvegicus) were subjected to bilateral laparotomy to remove the ovaries. The animals were divided into a control group, in which they only underwent laparotomy; an ovariectomized group (OVX); an ovariectomized group treated with 1 mg/kg of alendronate (OVX 1 mg); and an ovariectomized group treated with 2 mg/kg of alendronate (OVX 2 mg). The rats received alendronate twice a week for 90 days. The left femur was then removed, fixed and processed for embedding in paraffin. Semi-serial sections stained with hematoxylin and eosin were used to determine the area occupied by organic bone matrix, by means of image analysis software. The animals' weights were obtained at the beginning and end of the experiment.RESULTS: The ovariectomized animals and those treated with 1 mg/kg of alendronate presented significant increases in body weight (p < 0.05), in comparison with the control group. Histomorphometric analysis revealed that in the animals treated with 2 mg/kg of alendronate, the area (µm2) occupied by organic matrix (1,81,900 ± 18,130) was similar (p > 0.05) to that of the non-ovariectomized control animals (2,04,800 ± 9590), which indicates that this medication had a preventive effect with regard to bone mass loss.CONCLUSION: The higher concentration of the medication, administered twice a week for 90 days, was more effective than the dose of 1 mg/kg over the same period. OBJETIVO: avaliar a efeito de duas concentrações diferentes de alendronato de sódio (ALN) sobre a quantidade de matriz orgânica no fêmur de ratas com supressão estrogênica provocada por ovariectomia.MÉTODOS: ratas Wistar (Rattus norvegicus) com 60 dias foram submetidas a laparotomia bilateral para remoção dos ovários. Os animais foram divididos em grupo controle, no qual os animais foram apenas laparotomizados; grupo ovariectomizado (OVX); grupo ovariectomizado tratado com 1 mg/kg de ALN (OVX 1 mg); e grupo ovariectomizado tratado com 2 mg/kg de ALN (OVX 2 mg). Receberam ALN duas vezes por semana durante 90 dias. O fêmur esquerdo foi coletado, fixado e processado para inclusão em parafina. Cortes semisseriados corados com H&E foram usados para a obtenção, com um software de análise de imagens, da área ocupada por matriz óssea orgânica. O peso dos animais foi obtido no inicio e no fim do experimento.RESULTADOS: os animais ovariectomizados e aqueles tratados com 1 mg/kg de ALN tiveram um aumento significativo (p < 0,05) no peso corporal quando comparados com o grupo controle. A análise histomorfométrica revelou que nos animais tratados com 2 mg/kg de ALN a área (µm2) ocupada por matriz orgânica (181.900 ± 18.130) foi semelhante (p > 0,05) àquela dos animais controle não ovariectomizados (204.800 ± 9.590), o que indica um efeito preventivo desse medicamento sobre a perda de massa óssea.CONCLUSÃO: a maior concentração do medicamento administrado duas vezes por semana por 90 dias foi mais eficaz do que a dose de 1 mg/kg no mesmo período.

ResumoObjetivoavaliar a efeito de duas concentrações diferentes de alendronato de sódio (ALN) sobre a quantidade de matriz orgânica no fêmur de ratas com supressão estrogênica provocada por ovariectomia.Métodosratas Wistar (Rattus norvegicus) com 60 dias foram submetidas a laparotomia bilateral para remoção dos ovários. Os animais foram divididos em grupo controle, no qual os animais foram apenas laparotomizados; grupo ovariectomizado (OVX); grupo ovariectomizado tratado com 1 mg/kg de ALN (OVX 1 mg); e grupo ovariectomizado tratado com 2 mg/kg de ALN (OVX 2 mg). Receberam ALN duas vezes por semana durante 90 dias. O fêmur esquerdo foi coletado, fixado e processado para inclusão em parafina. Cortes semisseriados corados com H&E foram usados para a obtenção, com um software de análise de imagens, da área ocupada por matriz óssea orgânica. O peso dos animais foi obtido no inicio e no fim do experimento.Resultadosos animais ovariectomizados e aqueles tratados com 1mg/kg de ALN tiveram um aumento significativo (p0,05) àquela dos animais controle não ovariectomizados (204.800±9.590), o que indica um efeito preventivo desse medicamento sobre a perda de massa óssea.Conclusãoa maior concentração do medicamento administrado duas vezes por semana por 90 dias foi mais eficaz do que a dose de 1mg/kg no mesmo período.AbstractObjectiveto evaluate the effect of two different concentrations of sodium alendronate on the quantity of organic matrix in the femur of rats with estrogen suppression caused by ovariectomy.Methodssixty‐day‐old Wistar rats (Rattus norvegicus) were subjected to bilateral laparotomy to remove the ovaries. The animals were divided into a control group, in which they only underwent laparotomy; an ovariectomized group (OVX); an ovariectomized group treated with 1 mg/kg of alendronate (OVX 1 mg); and an ovariectomized group treated with 2 mg/kg of alendronate (OVX 2 mg). The rats received alendronate twice a week for 90 days. The left femur was then removed, fixed and processed for embedding in paraffin. Semi‐serial sections stained with hematoxylin and eosin were used to determine the area occupied by organic bone matrix, by means of image analysis software. The animals’ weights were obtained at the beginning and end of the experiment.Resultsthe ovariectomized animals and those treated with 1mg/kg of alendronate presented significant increases in body weight (p0.05) to that of the non‐ovariectomized control animals (204,800±9,590), which indicates that this medication had a preventive effect with regard to bone mass loss.Conclusionthe higher concentration of the medication, administered twice a week for 90 days, was more effective than the dose of 1mg/kg over the same period.

Fusarium is an emerging pathogen with high levels of morbidity and mortality. A significant tissue response is observed in infected patients, and the condition has been associated with the production of toxic metabolites. The aim of the present study was to identify a major fraction of crude metabolic extract of Fusarium oxysporum and investigate its effects on the skin of healthy rats. Fraction F1 was obtained from the cultivation of F. oxysporum in Czapek–Dox. In the treatment groups, fraction F1 (0.05 mg/ ml) was injected intradermally, while (50 µl) 0.9% of saline solution was injected in the control groups. The animals were killed 3, 6, 12, and 24 h after inoculation. The skin was fixed for inclusion in paraffin to obtain histological sections and stained with hematoxylin and eosin, Sirius red, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling. Samples were analyzed using Fourier transform Raman spectroscopy. The tissue reactions were classified and compared over time and by treatment. In the treatment group, inflammatory reaction peaked at 6 h, being classified as moderate, with infiltrate composed mainly of neutrophils. terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining was negative. The area occupied by types I and III collagen in the treatment group increased over time. There was a change in the area occupied by amide I and the ratios of the –CH2 and –CH3 molecules. It can be argued that the fraction F1 contains elements that contribute to the invasion of Fusarium in the skin, destructurizing the organization of the extracellular matrix. Copyright © 2014 John Wiley & Sons, Ltd.

This study evaluated physical, chemical and morphological changes in lungs of mice infected with Paracoccidioides brasiliensis. The animals were inoculated with 0.1 mL of fungal suspension of the P. brasiliensis 18 isolate and were euthanized 1, 2, 4 and 8 weeks after inoculation. The upper left lobe of the lung was isolated, fixed and processed for paraffin embedding. The sections were stained with H&E for histopathological study, with Gomori-Grocott to locate and identify the fungus, and with TUNEL immunostaining to detect the occurrence of programmed cell death. The lower and middle right lobes were analyzed by Fourier Transform Infrared Photoacoustic Spectrocopy (FTIR-PAS) to investigate physical and chemical features of the infected lungs. The results showed that lungs infected by P. brasiliensis underwent structural changes that varied according to the time period analyzed, and that changes in the absorption bands of different chemical groups resulted from these morphological changes. The results suggest that the combination of FTIR-PAS spectroscopy with morphological evaluation is an effective procedure for the study of paracoccidioidomycosis, one of the most important systemic mycoses that can damage the lung architecture and consequently impair the respiratory function.

Creatine is widely used by athletes as an ergogenic resource. The aim of this study was to evaluate the infl uence of creatine supplementation on the duodenum of rats submitted to physical training. The number and myenteric neuronal cell bodies as well mucosal and muscular tunic morphometry were evaluated. Control animals received a standard chow for 8 weeks, and the treated ones received the standard chow for 4 weeks and were later fed with the same chow but added with 2% creatine. Animals were divided in groups: sedentary, sedentary supplemented with creatine, trained and trained supplemented with creatine. The training consisted in treadmill running for 8 weeks. Duodenal samples were either processed for whole mount preparations or for paraffi n embedding and hematoxylin-eosin staining for histological and morphometric studies of the mucosa, the muscular tunic and myenteric neurons. It was observed that neither creatine nor physical training alone promoted alterations in muscular tunic thickness, villus height or crypts depth, however, a reduction in these parameters was observed when both were associated. The number of myenteric neurons was unchanged, but the neuronal cell body area was reduced in trained animals but not when training and creatine was associated, suggesting a neuroprotector role of this substance

Introduction Skin lesions are a significant public health problem, above all that wounds fail to heal properly and become chronic. Due to its reepithelization action, insulin has the potential to heal skin lesions, by stimulating the proliferation and migration of keratinocytes, angiogenic stimulus, and increasing collagen deposition. In the present study insulin was complexed with 2-hydroxypropyl-β-cyclodextrin (HPβCD) and its wound healing effect and inclusion complex (HPβCD-I) were evaluated in excisional wounds in the skin of rats. Material and methods Three different gel based pharmaceutical forms were created: carbopol 940® base gel, an insulin gel comprising the base gel plus 50 IU of insulin and a gel complex comprising the base gel plus (HPβCD) complexed with insulin (HPβCD-I) were used to verify wound healing in vitro and in vivo assays. Results The wounds in the skin of rats were treated with gel containing HPβCD-I not cytoxically irritating and cytotoxic. Analysis of cell proliferation and measurement of the length and thickness of the epidermis showed that HPβCD-I prolonged the proliferation and migration of keratinocytes. Revascularization analysis of lesions treated with HPβCD-I compared to those treated with insulin found that angiogenic stimulus was less intense, but more constant and prolonged in the modified release process. There was increased deposition of type I and III collagen fibers in accordance with the treatment time. Conclusion Therefore, the slow release of complexed insulin modulated the reepithelialization process by stimulating cell proliferation and migration of keratinocytes, favoring greater concentration of serum insulin, modulating inflammatory response, matrix remodeling and promoting neovascularization. Angiogenesis extended by the steady release of insulin can be effective in the treatment of chronic wounds.

The effects of starter diet (days 1 to 21) supplemented with arginine (Arg) on the production performance and duodenum and jejunum mucosa morphometry of broilers were studied. Male Cobb broiler chickens (990) were randomly assigned to one of five treatments in a complete random design. Measurements of 33 chicks per treatment were made in six repetitions. The treatments consisted of a basal diet with 1.390% digestible Arg (no supplementation) and four dietary levels (1.490%, 1.590%, 1.690%, and 1.790%), providing a relationship with lysine of 1.103; 1.183; 1.262; 1.341 and 1.421%, respectively. From the age of 22 days on, all birds received conventional grower diet. The data were submitted to regression analysis by polynomial decomposition of the degrees of freedom in relation to the levels of Arg. The Arg supplementation increased (P0.05) in the growth phase (days 22 to 42) in the absence of the Arg supplementation. The supplementation of Arg over of NRC recommendation during the starter phase may be necessary for the expression of the maximal weight gain potential in birds. No effect (P0,05) na fase de crescimento (22 a 42 dias) na ausência de suplementação de Arg. A suplementação acima do recomendado pelo NRC na fase inicial pode ser necessária para a expressão máxima do potencial de ganho de peso em aves. Não houve efeito da suplementação de Arg na dieta no peso e comprimento do intestino delgado em nenhuma idade. No entanto, a relação vilo:cripta no duodeno aumentou e a profundidade da cripta diminuiu na primeira semana em resposta ao incremento de Arg na dieta. Concluiu-se que em frangos de corte a suplementação de Arg na dieta inicial melhorou o desempenho e a morfometria do intestino delgado, especialmente na primeira semana.

All methods to detect experimental loss of bone present technique limitations. The sensitivities of image and histological analyses to detect the effects of teriparatide in rats with bone loss after ovariectomy were evaluated. All methods were qualitatively valid. The standardization of methods to assess bone loss after ovariectomy is crucial to establish the degree of experimental osteoporosis. In general, methods per image or histological techniques are used. To validate these two ways to determine the degree of bone loss in ovariectomized rats, we evaluated the sensitivities of bone densitometry, conventional radiography, and histological analysis of the area occupied by collagen, detecting the effects of teriparatide treatment in the femur of ovariectomized rats with bone loss. Wistar rats were divided into three groups: a control group, in which the animals were only subjected to laparotomy; an ovariectomized group, in which bilateral removal of the ovaries was performed; and an ovariectomized + teriparatide group, in which bilateral removal of the ovaries was performed, and the animals were treated with 3 μg/100 g/day of teriparatide. Three months following the ovariectomy, bone densitometry, radiographic densitometry, and histological analysis of the area occupied by collagen fibers were carried out in the femur diaphysis. The bone densitometry revealed 11.2% reduction in femur density; in the conventional radiography, the loss of bone mass was 14.5%, and with the histological analysis, a 40.9% reduction in the area occupied by collagen was detected in the femur diaphysis. In conclusion, histological analysis could not be quantitatively compared with the methods of bone densitometry and conventional radiography; however, all of these methods were qualitatively valid for assessing the loss of bone stemming from ovariectomy and the therapeutic effect of teriparatide in the prevention of osteoporosis.

Background Most patients with Fusarium infection present with affection of the tegumentary system, with the presence of necrotic and/or inflammatory lesions associated with pain. Aim To evaluate the effect of the intradermal application of a metabolic extract of Fusarium oxysporum in the skin of Wistar rats. Methods The extract was obtained from fungal cultivation in Czapek-Dox medium. It was sterilized by filtration through a Millipore membrane, and injected (0.5 mg/mL) intradermally into the skin of rats, which were killed 3, 6, 12, and 24 h after inoculation. Skin specimens were placed in paraffin and stained using hematoxylin and eosin, and toluidine blue, for the evaluation of the inflammatory response, and Sirius red for the quantification of collagen. Terminal deoxynucleotidyl transferase-mediated UTP nick end labelling (TUNEL) was used for the identification of apoptosis. Tissue reactions were graded and compared over time and compartment. Results The inflammatory reaction reached a peak at 12 h for both the dermis and subcutaneous region, being graded as moderate and moderate to severe, respectively. There was an influx of granulocytes, lymphocytes, and macrophages. A significant increase in the number of mast cells, as well as the presence of hyperemic vessels and apoptotic bodies, was observed. There was TUNEL staining in keratinocytes, fibroblasts, endothelial cells, muscles, and in the cells of the inflammatory infiltrate. There was a significant decrease in the area occupied by collagen after 12 h. Conclusions The extract induced histopathologic alterations in the skin, probably as a result of the presence of active toxic metabolites.

A morphologic study of the lungs was carried out in Swiss mice infected with yeast isolated from Paracoccidioides brasiliensis (Pb18). The lung was processed 1, 2, 4, and 8 weeks after inoculation for histologic staining with hematoxylin and eosin (H&E), methenamine silver nitrate (Gomori-Grocott), and picrosirius to qualitative and quantitative analyses of the granulomas and the presence of fungal lesions. The numbers of CFUs/g counted in the lungs were 189.8 +/- 20.64, 353.6 +/- 46.21, 547.2 +/- 108.1, and 295.2 +/- 89.17 in the first, second, fourth, and eighth weeks, respectively. One week after infection, inflammatory cells and reticular and collagens fibers, the latest typical of fibrosis, were detected. After 2 and 4 weeks, a progressive intensification of the infection and fibrosis was observed, but in week 8 a more organized granuloma was evident, with macrophages, epithelioid cells, and yeasts in the central portion, and intense peripheral basophilia. Pycnotic structures typical of apoptotic bodies were observed in weeks 1 and 8. The different histologic staining used acted as a fundamental tool for the study of the morphologic organization of granuloma formation.

Our aim was to study the ultrastructural aspects in the genesis of blood capillaries in the lower limb skeletal muscle of rats submitted to ischemia under the action of intramuscular or endovenous prostaglandin E(1) (PGE(1)). Twelve Wistar-UEM rats were used, randomly distributed into three groups of four animals each, equally redistributed into two subgroups, observed at 7 and 14 days as follows: group only with ischemia was considered as control (I), group with ischemia and intramuscular injection of PGE(1) (IM), and group with ischemia and endovenous injection of PGE(1) (EV). Results were analyzed by transmission electronic microscopy (TEM). TEM analysis revealed evidence of new capillary formation. TEM permitted us to identify morphological structures and phenomena in vascular neoformation that might have occurred through angiogenesis and/or vasculogenesis.

Notre but etait d'etudier les aspects ultrastructuraux de la genese des capillaires sanguins dans le muscle squelettique des membres inferieurs de rats soumis a une ischemie sous l'action de la prostaglandine E 1 (PGE 1 ) intra-musculaire ou intra-veineuse. Douze rats Wistar-UEM ont ete utilises, distribues de facon randomisee en trois groupes de quatre animaux chacun, egalement redistribues en deux sous-groupes, observes a 7 et 14 jours comme suit : groupe I avec ischemie seule, considere comme groupe controle, groupe avec ischemie et injection intra-musculaire de PGE 1 (groupe IM) et groupe avec ischemie et injection intra-veineuse de PGE 1 (groupe IV). Les resultats ont ete analyses en microscopie electronique de transmission (MET). Les analyses en MET ont montre la formation de neocapillaires. La MET nous a permis d'identifier les structures et les phenomenes morphologiques de la neoformation vasculaire qui auraient pu survenir par angiogenese et/ou vasculogenese.

CONTEXTO: A angiogênese terapêutica é uma modalidade de tratamento para pacientes com insuficiência arterial crônica que não têm indicação para revascularização direta ou angioplastia e que não tiveram uma resposta satisfatória ao tratamento clínico. Entre as drogas utilizadas para essa finalidade está a prostaglandina E1 (PGE1). OBJETIVO: Estudar os aspectos morfológicos na gênese de capilares sanguíneos em músculo esquelético do membro caudal de ratos submetidos à isquemia sob a ação da PGE1, administrada por via intramuscular (IM) ou endovenosa (EV). MÉTODOS: Foram utilizados 48 ratos, linhagem Wistar-UEM, distribuídos aleatoriamente em três grupos de 16, redistribuídos igualmente em dois subgrupos, observados no 7º e 14º dias, sendo um grupo controle onde apenas foi provocada a isquemia no membro, outro com a isquemia e a injeção da PGE1 via IM e outro com a isquemia e a injeção da PGE1 EV. Para análise dos resultados, foram realizadas a coloração com hematoxilina e eosina (HE) e coloração imuno-histoquímica. RESULTADOS: Constatou-se um aumento estatisticamente significativo no número de capilares nos subgrupos com o uso da PGE1 IM e EV, através da contagem nos cortes corados com HE. A imunomarcação não foi eficiente para a quantificação dos capilares. CONCLUSÕES: A PGE1, administrada por via IM ou EV, promoveu, após 14 dias de observação, um aumento no número de capilares no músculo esquelético de ratos submetido à isquemia, identificáveis histologicamente com a coloração em HE. A imunocoloração não permitiu estabelecer uma correlação com o aumento de vasos encontrados na coloração com HE. BACKGROUND: Therapeutic angiogenesis is a treatment modality for patients with chronic arterial insufficiency who do not have indication for direct reconstruction or angioplasty and who were not successfully submitted to clinical treatment. Prostaglandin E1 (PGE1) is one of the drugs used for this purpose. OBJECTIVE: To study morphologic aspects in the genesis of blood capillaries in the lower limb skeletal muscle of rats submitted to ischemia under the action of intramuscular (IM) or endovenous (EV) PGE1. METHODS: Forty-eight Wistar-UEM rats were randomly distributed into three groups of 16, equally redistributed into two subgroups, observed at the 7th and 14th days as follows: one control group, which had only limb ischemia; one group with ischemia and IM injection of PGE1; and one group with ischemia and EV injection of PGE1. To analyze the results, hematoxylin-eosin (HE) and immunohistochemical staining were used. RESULTS: There was a statistically significant increase in the number of capillaries in the subgroups using IM or EV PGE1, through counting in the samples containing HE staining. Immunostaining was not efficient for the quantification of capillaries. CONCLUSIONS: IM or EV PGE1 resulted in an increase in number of capillaries in the skeletal muscle of rats submitted to ischemia after 14 days of observation, which was histologically identifiable through HE staining. Immunostaining was not successful in establishing a correlation with the increase in vessels found in HE staining.

Introduction: The topical and intradermal photodynamic therapy (PDT) effect of methylene blue (MB) using light-emitting diode (LED) as light source (MB/LED-PDT) in the treatment of lesions of American cutaneous leishmaniasis (ACL) caused by Leishmania braziliensis in hamsters were investigated. Methods: Hamsters were infected in the footpad with 4×10 7 promastigotes of L. braziliensis and divided in 4 groups: Control group was not treated, AmB group was treated with amphotericin B, MB-Id group received intradermal MB at the edge of the lesion and MB-Tp group received MB topic. After treatment with MB, the animals were illuminated using red LEDs at the 655 nm wavelength for 1 hour. The MB/LED-PDT was carried out three times a week for 12 weeks. Results: Animals of MB-Tp group presented lesion healing with significant diminution in extent of the lesion, and reduced parasite burden compared to control group; however, no significant difference was seen compared to the AmB group. MB-Tp group also showed reconstitution of the epithelium, the formation of collagen fibers, organization in the epidermis, a little disorganization and inflammation in the dermis. MB-Id was ineffective in all parameters evaluated, and it was comparable to the control group results. Conclusion: These data show that PDT with the use of MB-Tp and LED may be an alternative for the treatment of ACL. However, additional studies are being conducted to assess the potential of MB/LED-PDT, alone or in combination with conventional therapy, for the treatment of ACL.

The enteric nervous and enteroendocrine systems regulate different processes in the small intestine. Ablation of myenteric plexus with benzalkonium chloride (BAC) stimulates epithelial cell proliferation, whereas endocrine serotonin cells may inhibit the process. To evaluate the connection between the systems and the influence of myenteric plexus on serotoninergic cells in rats during postnatal development, the ileal plexus was partially removed with BAC. Rats were treated at 13 or 21 days and sacrificed after 15 days. The cell bodies of myenteric neurons were stained by beta NADH-diaphorase to detect the extension of denervation. The number of enteroendocrine cells in the ileum was estimated in crypts and villi in paraffin sections immunostained for serotonin. The number of neurons was reduced by 27.6 and 45% in rats treated on the 13th and 21st days, respectively. We tried to establish a correlation of denervation and the serotonin population according to the age of treatment. We observed a reduction of immunolabelled cells in the crypts of rats treated at 13 days, whereas this effect was seen in the villi of rats denervated at 21 days. These results suggest that the enteric nervous system might control the enteroendocrine cell population and this complex mechanism could be correlated to changes in cell proliferation.

The objective of the present study was to investigate the effects of monotherapy with simvastatin or ezetimibe compared with those of an ezetimibe + simvastatin combination on the histopathologic aspects of arthritis induced by Complete Freund’s Adjuvant in rats. The characteristics of diarthrosis were analyzed 7 and 28 days after arthritis induction with regard to the regularity and integrity of articular cartilage, the presence of leukocytes in blood vessels at the inflammation site, and the cytokine levels in articular tissue. Monotherapy with simvastatin or ezetimibe and the ezetimibe + simvastatin combination effectively reduced the cytokine levels (interleukin-6 and tumor necrosis factor) and articular lesions that are characteristic of this experimental disease. Although the results did not reveal significant differences between the monotherapy and the combined therapy, they suggest that these drugs have considerable antiinflammatory properties, as reflected by an attenuation of articular cartilage lesions mediated by a reduction in the levels of proinflammatory cytokines.

Paracoccidioidomycosis is the most important systemic mycosis in Latin America. The main entrance of the fungus is the airway. It primarily occurs in the lung, but in its disseminated form may affect any organ. The liver is one of the organs afflicted by this disease and its homeostasis may be impaired. The aim of the present study was to evaluate the evolution of paracoccidioidomycosis in the liver of Swiss mice and correlate morphological factors with the expression of gp43 and with physicochemical analysis via FT-Raman of the infected organ. According to colony forming unit (CFU) and granuloma counting, the first and second weeks were the periods when infection was most severe. Tissue response was characterized by the development of organized granulomas and widespread infection, with yeasts located within the macrophages and isolated hepatocytes. The gp43 molecule was distributed throughout the hepatic parenchyma, and immunostaining was constant in all observed periods. The main physicochemical changes of the infected liver were observed in the spectral ranges between 1700-1530 cm(-1) and 1370-1290 cm(-1), a peak shifting center attributed to phenylalanine and area variation of -CH2 and -CH3 compounds associated to collagen, respectively. Over time, there was a direct proportional relationship between the number of CFUs, the number of granulomas and the physicochemical changes in the liver of mice infected with Paracoccidioides brasiliensis. The expression of gp43 was similar in all observed periods.

Acute liver damage caused by acetaminophen overdose is a significant clinical problem and could benefit from new therapeutic strategies.Objective. This study investigated the hepatoprotective effect ofThymus vulgarisessential oil (TEO), which is used popularly for various beneficial effects, such as its antiseptic, carminative, and antimicrobial effects. The hepatoprotective activity of TEO was determined by assessing serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) in mice. Their livers were then used to determine myeloperoxidase (MPO) enzyme activity and subjected to histological analysis. In vitro antioxidant activity was evaluated by assessing the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH•)-scavenging effects of TEO and TEO-induced lipid peroxidation. TEO reduced the levels of the serum marker enzymes AST, ALT, and ALP and MPO activity. The histopathological analysis indicated that TEO prevented acetaminophen-induced necrosis. The essential oil also exhibited antioxidant activity, reflected by its DPPH radical-scavenging effects and in the lipid peroxidation assay. These results suggest that TEO has hepatoprotective effects on acetaminophen-induced hepatic damage in mice.

This study focused on the therapeutic effect of a propolis SLNC 106PIextract on experimental colitis. Wistar adult rats received 0.8 mL rectal dose of one of the following solutions: saline (group S), 20 mg TNBS in 50% ethanol (group TNBS), 20 mg TNBS in 50% ethanol and propolis extract in saline (group TNBS-P), propolis extract in saline (group SP), and 20 mg TNBS in 50% ethanol and 50 mg/kg mesalazine (group TNBS-M). The animals were euthanized 7 or 14 days after the colitis induction. Samples of the distal colon were harvested for the analysis of myeloperoxidase (MPO) enzyme activity and for morphometric analysis in paraffin-embedded histological sections with hematoxylin-eosin or histochemical staining. The animals treated with TNBS exhibited the typical clinical signs of colitis. Increased MPO activity confirmed the presence of inflammation. TNBS induced the development of megacolon, ulceration, transmural inflammatory infiltrate, and thickened bowel walls. Treatment with propolis moderately reduced the inflammatory response, decreased the number of cysts and abscesses, inhibited epithelial proliferation, and increased the number of goblet cells. The anti-inflammatory activity of the propolis SLNC 106 extract was confirmed by the reductions in both the inflammatory infiltrate and the number of cysts and abscesses in the colon mucosa.

To apply the photoacoustic spectroscopy technique to investigate the penetration rate of topically applied Helicteres gardneriana extract used as anti-inflammatory agent.Experiments were performed ex vivo in a well-controlled group of mice. The crude extract was obtained from leaves of the plant Helicteres gardneriana. Croton oil was applied into the ventral surface of the mouse's right and left auricles in order to induce an inflammatory response. The left auricle was treated with crude extract, while the right one served as the control. After 6 h, the auricles were sectioned for measurements of edema intensity, myeloperoxidase activity and the formulation penetration rate.Croton oil induced inflammatory response in both auricles. The application of Helicteres gardneriana extract reduced significantly the edema of the auricle and inhibited the activity of the myeloperoxidase enzyme. The photoacoustic data showed that the propagation of the formulation was efficient to reach the deep region of the auricle, crossing the cartilage. The strong anti-inflammatory effect was associated with the observed deep penetration of the formulation.This pre-clinical study showed the anti-inflammatory effect of Helicteres gardneriana extract. The photoacoustic technique was useful to demonstrate that this anti-inflammatory activity was associated with deep percutaneous penetration.

Experimental models of inflammatory bowel disease (IBD) supply considerable information on the pathogenesis of this illness and represent an important tool in testing new strategies of treatment. Crohn’s disease (CD) is an idiopathic, chronic inflammatory condition of the intestinal tract that causes structural and functional changes in the enteric nervous system (ENS), and for which therapeutics options are limited. The use of natural products as alternative therapy for CD has been intensively investigated in animal models of IBD. This review summarizes the major structural abnormalities of the ENS in animal models of Crohn’s disease and the effects of natural products in the inflammatory process. The bibliographical research, of descriptive character, comprises a period of 10 years and was carried through consultation of printed materials and the database of Literatura Latino Americana em Ciencias da Saude e do Caribe (LILACS) and in the National Library of Medicine (MEDLINE), displayed in electronic support. Books have also been used.

The aim of present study was to evaluate the number and basophily of cell bodies of myenteric neurons in the ileum of rats with diabetes mellitus induced by streptozotocin. Four groups of rats were used: diabetes was induced in two (D) whereas the other two worked as controls (N). Animals were sacrificed six (6N, 6D) or nineteen (19N, 19D) weeks after diabetes induction. A segment of the terminal portion of the ileum of each rat was obtained and stained with Giemsa's solution, for whole-mount preparation studies. Forty fields were analyzed in each animal, and the number and basophily intensity of cell bodies were recorded. After counting, the following mean numbers of neurons/mm² were obtained: 6N=593.1 ± 95.75, 6D=639.1 ± 130.8, 19N=580.1 ± 175.6 and 19D=402.0 ± 144.8. The analysis of basophily shown that highest frequency of neurons with weak/intermediary basophily was verified in 6D group (55.3%), whereas the groups 6N, 19N e 19D presented 38%, 36% e 40% respectively. The statistical analysis showed that a long period is necessary to decrease the number of neurons/mm² in the rat ileum after diabetes induction, and that there was a reduction in basophily intensity in diabetic rats after 6 weeks of treatment, and such cells do not recover after a longer period (19 weeks). O objetivo deste estudo foi avaliar o número e basofilia dos corpos celulares dos neurônios mientéricos no íleo de ratos com diabetes mellitus induzido por estreptozootocina. Quatro grupos de ratos foram usados. O diabetes foi induzido em dois grupos (D), enquanto outros dois eram controles (N). Os animais foram sacrificados 6 (6N, 6D) ou dezenove (19N, 19D) semanas após a indução do diabetes. Preparados totais de um segmento do íleo terminal, de cada rato, foram corados com solução de Giemsa. Foram contados 40 campos em cada animal, e o número e a intensidade de basofilia citoplasmática foram registrados. Após a contagem, as seguintes médias no número de neurônios/mm² foram obtidos: 6N=593,1 ± 95,75, 6D=693,1 ± 130,8, 19N=580,1 ± 175,6 e 19D=402,0 ± 144,8. A análise da basofilia mostrou que a maior frequência de neurônios com basofilia fraca e intermediária foi verificada no grupo 6D (55,3%), enquanto os grupos 6N, 19N e 19D apresentaram 38%, 36% e 40% respectivamente. A análise estatística mostrou que um longo período é necessário para que ocorra a redução no número de neurônios/mm² no íleo de ratos, após a indução do diabetes. Também demonstrou uma redução na intensidade da basofilia citoplasmática 6 semanas de tratamento com estreptozootocina, e que estas células não se recuperam após um longo período de tempo (19 semanas).

ObjectivesThe aim of the present study was to evaluate the effect of two different dosages of alendronate on induced orthodontic movement in an experimental model involving rats with osteoporosis following ovariectomy.DesignFemale Wistar rats (Rattus norvegicus) eight weeks of age were divided into four groups (n=12/group): ovariectomized (OVX group); ovariectomized and treated with alendronate sodium at 1mg/kg (Group OVX+ALN1); ovariectomized and treated with alendronate sodium at 2mg/kg (Group OVX+ALN2); and sham operated (control). Three months after ovariectomy, the maxillary right first molar was submitted to movement for five and seven days. After the death of the animals, the maxilla were removed and processed for microscopic evaluation. The maxillary left first molar (without movement) was used for comparison purposes in all groups. The samples were processed for the quantification of alveolar bone and tooth movement.ResultsIntragroup comparisons showed significant movement after five and seven days (p0.05). Movement in ovariectomized+alendronate groups were also smaller than non-ovariectomized rats, however without statistical difference.

This study evaluated the cicatrizant effect of an ointment containing 1% of the ethyl-acetate fraction extracted from the stem bark of "barbatimão" (Stryphnodendron adstringens), in wounds made in the skin of rats, after 4, 7 and 10 days of treatment. Control wounds were treated with a base ointment without extract. The proliferation of keratinocytes in the area of reepithelialization was evaluated by counting the number of epithelial cells that were blocked in metaphase by vincristine sulfate. The length of the reepithelialized margin and the contraction of the wound were measured. Topical application of the "barbatimão" ointment stimulated proliferation of the keratinocytes, but had no effect on the length of the epithelium or on the contraction of the wounds.Neste estudo, avaliou-se a atividade cicatrizante de uma pomada contendo uma fração acetato de etila 1% obtida de cascas de "barbatimão" (Stryphnodendron adstringens) em feridas excisionais na pele de ratos após 4, 7 e 10 dias de tratamento. Feridas controle foram tratadas com pomada base, sem extrato. A proliferação dos queratinócitos na área reepitelizada foi avaliada através da contagem do número de queratinócitos bloqueados em metáfase, pelo sulfato de vincristina. O comprimento da margem reepitelizada e a contração das feridas foram mensurados. As feridas tratadas com barbatimão apresentaram um maior número de mitoses do que aquelas tratadas com a pomada base, em todos os tempos avaliados. A aplicação tópica da pomada de "barbatimão" estimulou a proliferação epitelial contudo não teve efeito sobre a migração dos queratinócitos ou sobre a contração das feridas.