Oxylipin profiling in endothelial cells in vitro - Effects of DHA and hydrocortisone upon an inflammatory challenge.

Omega-3 poly-unsaturated fatty acids have been shown to have beneficial effects on several inflammatory-driven endpoints such as cardiovascular diseases. The anti-inflammatory effects of docosahexaenoic acid (DHA) are largely mediated through various oxylipins. Yet, mechanistic insights are limited. Here, we measured 53 oxylipins using LC-MS/MS in an in vitro model of endothelial cell inflammation, and compared the changes induced by DHA to hydrocortisone, a well-established anti-inflammatory drug. DHA modified several oxylipins derived from different precursors such as DHA, AA, LA and EPA. In response to a TNFα and IL-1-ß challenge, DHA clearly reduced many COX-derived pro-inflammatory oxylipins, yet to a minor extent when compared to hydrocortisone. DHA also upregulated metabolites from the CYP and LOX pathways as opposed to hydrocortisone. Thus, DHA reduced pro-inflammation and enhanced pro-resolution, while hydrocortisone blunted both the pro- and anti-inflammatory pathways. Our results may fuel further research on the mitigation of corticosteroids adverse side-effects.

Ultrasonic shears and bipolar vessel-sealing system for three-port laparoscopic ovariohysterectomy in dogs / Bisturi ultrassônico e sistema de selamento de vasos sanguíneos para ovário-histerectomia laparoscópica com três portais em cães

In the current experimental study, two different instruments were compared for prophylactic hemostasis during a 3-port technique of laparoscopic ovariohysterectomy (OVH) in bitches. Moreover, the need for port enlargement for specimen retrieval using 5mm or 10mm trocartes, technical difficulties and complications were assessed. Ultrasonic energy and a vascular sealing system were tested. Duration of surgery, patient weight and diameter of the uterine arteries and veins were compared among the groups. Fifteen dogs randomly divided into three groups (GI, GII, GIII) were submitted to ovariohysterectomy with hemostasis from the use of ultrasonic scalpel or vascular sealing equipment. In GI and GII the LigaSure(tm) impedance-controlled bipolar vessel-sealing device was used, differing in the size of trocartes used between groups, and in GIII Autosonix(tm) was used. Patients' weight, duration of surgery, technical difficulties and complications were compared among the groups. Statistical analysis showed no difference between the data analyzed among the groups. In conclusion, both methods of hemostasis were effective for 3-port total laparoscopic ovariohysterectomy in bitches. However, 5mm trocartes were not adequate for retrieval of the uterus and ovaries without need for enlargement of the port incision.

No presente experimento, foram comparados dois diferentes equipamentos, um com emprego de energia ultrassônica, e outro utilizando sistema de selamento vascular, como única forma de hemostasia em ovário-histerectomias laparoscópicas com três portais em cadelas. Ademais, a necessidade de ampliação da incisão do portal de 5mm ou de 10mm para a remoção dos espécimes ressecados, dificuldades técnicas e complicações foram avaliadas. Quinze cadelas aleatoriamente distribuídas em três grupos (GI, GII, GIII) foram submetidas à ovário-histerectomia com hemostasia a partir da utilização de bisturi ultrassônico ou equipamento de selamento vascular. Nos GI e GII foi utilizado um sistema de selamento vascular eletrotérmico bipolar (LigaSure(r)), diferindo-se no tamanho dos portais utilizados; em GIII foi utilizado bisturi ultrassônico (Autosonix(r)). A análise estatística não demonstrou diferença significativa entre os dados analisados nos três grupos. Conclui-se que ambos os métodos de hemostasia utilizados mostraram-se adequados e viabilizaram a realização de ovário-histerectomia laparoscópica em cadelas. Todavia, o trocarte de 5mm não foi adequado para remoção do útero e ovários sem necessidade de ampliação da incisão de acesso.

Avaliação de dieta formulada com subprodutos de mandioca na produção de sêmen de coelhos / Evaluation of diet formulated with cassava by-products in the rabbit semen production

Avaliaram-se as características quantitativas e qualitativas do sêmen de coelhos alimentados com uma ração referência e outra ração formulada com 79,83% de subprodutos de mandioca. Utilizaram-se 20 reprodutores da raça Nova Zelândia Branco, com idade média inicial de sete meses, alojados individualmente em gaiolas e distribuídos em delineamento experimental inteiramente ao acaso, com duas dietas e dez repetições. Realizaram-se dez colheitas de sêmen por animal durante seis meses. Avaliaram-se o volume de sêmen sem gel e volume de gel, cor do sêmen, pH, motilidade espermática progressiva, vigor espermático, concentração espermática, número de espermatozoides no ejaculado, espermatozoides normais, anormais e anormalidades primárias e secundárias. O volume de sêmen sem gel, o índice de normalidade, as anormalidades primárias e secundárias foram iguais (P>0,05) para os animais alimentados com a ração referência e a com subprodutos de mandioca. Os demais parâmetros do sêmen foram melhores (P<0,05) nos animais tratados com a ração com subprodutos de mandioca. Os resultados das características quali-quantitativas do sêmen dos coelhos da Raça Nova Zelândia Branco demonstram que é possível utilizar ração com 79,83% do volume da formulação com subprodutos de mandioca, na dieta de reprodutores, sem prejuízos nas características do sêmen, desde que observados os níveis reduzidos de taninos e ácido cianídrico.

The quantitative and qualitative rabbit semen characteristics fed with a reference diet and a diet formulated with 79.83% of cassava by-products were evaluated. 20 New Zealand White rabbit bucks, with initial average age of seven months were used, individually allocated and distributed in a completely ramdomized design, with two diets (treatments) and ten replications. The ten semen collections per animal were taken during six months. The evaluated parameters were: semen volume without gel and gel volume, color of semen, pH, spermatic progressive motility, spermatic vigor, spermatic concentration, spermatozoa number in the ejaculation, normal spermatozoa, abnormal and primary and secondary abnormalities spermatozoa. The semen volume without gel, the normal spermatozoa, primary and secondary abnormalities spermatozoa were similar (P>0.05) to the animals fed with cassava by-products diet and reference diet. Other semen parameters were better (P<0.05) in the animals fed with the cassava by-products diet. The results of the quali-quantitative characteristics of the semen from New Zealand White rabbits showed that it is possible to use diets containing 79.83% of inclusion of cassava by-products in the diet of rabbit bucks, without affecting semen characteristics, since we observed the reduced level of tannins and hydrocyanic acid.

The impact of adiposity on adipose tissue-resident lymphocyte activation in humans.

BACKGROUND/OBJECTIVES: The presence of T lymphocytes in human adipose tissue has only recently been demonstrated and relatively little is known of their potential relevance in the development of obesity-related diseases. We aimed to further characterise these cells and in particular to investigate how they interact with modestly increased levels of adiposity typical of common overweight and obesity. SUBJECTS/METHODS: Subcutaneous adipose tissue and fasting blood samples were obtained from healthy males aged 35-55 years with waist circumferences in lean (<94 cm), overweight (94-102 cm) and obese (>102 cm) categories. Adipose tissue-resident CD4+ and CD8+ T lymphocytes together with macrophages were identified by gene expression and flow cytometry. T lymphocytes were further characterised by their expression of activation markers CD25 and CD69. Adipose tissue inflammation was investigated using gene expression analysis and tissue culture. RESULTS: Participants reflected a range of adiposity from lean to class I obesity. Expression of CD4 (T-helper cells) and CD68 (macrophage), as well as FOXP3 RNA transcripts, was elevated in subcutaneous adipose tissue with increased levels of adiposity (P<0.001, P<0.001 and P=0.018, respectively). Flow cytometry revealed significant correlations between waist circumference and levels of CD25 and CD69 expression per cell on activated adipose tissue-resident CD4+ and CD8+ T lymphocytes (P-values ranging from 0.053 to <0.001). No such relationships were found with blood T lymphocytes. This increased T lymphocyte activation was related to increased expression and secretion of various pro- and anti-inflammatory cytokines from subcutaneous whole adipose tissue explants. CONCLUSIONS: This is the first study to demonstrate that even modest levels of overweight/obesity elicit modifications in adipose tissue immune function. Our results underscore the importance of T lymphocytes during adipose tissue expansion, and the presence of potential compensatory mechanisms that may work to counteract adipose tissue inflammation, possibly through an increased number of T-regulatory cells.

Protease inhibitor and metabolic alteration / Inhibidor de la proteasa y alteraciones metabólicas

Protease inhibitors (PIs), part of HAART (Highly Active Antiretroviral Therap) are selective, competitive inhibitors of protease, a crucial enzyme to viral maturation, infection and replication. A lipodystrophic syndrome has been reported in individuals treated with HAART, and associated to hyperglycemia, hypercholesterolemia, hypertrigliceridemia, hyperlipidemia, hypertension and hypreinsulinemia. The HAART-associated metabolic abnormalities were first associated with protease inhibitors, Ritonavir mostly, but the mechamisns that underlie these metabolic alterations are to date, not completely understood. Since PIs are candidate to be the drug of choice for other diseases treatment, such as the Hepatitis C, malaria and some types of cancer, it seems to be important to clarify the metabolic alterations associated to PIs. Wistar rats were treated twice a week with 30mg/kg Ritonavir for 4 and 8 weeks. Total cholesterol, HDL, LDL, VLDL, triglycerides and glycemic levels were measured by the end of each period of time selected. To avoid confunding effects of food intake, the animals were fasted 16 hours before. Our results showed rapid increase in serum triglycerides, total cholesterol, LDL-C and glycemic levels. No significant differences were observed for HDL-C or VLDL serum levels. Our study addresses the importance to observe the possible family history of dyslipidemia or diabetes, and control any other cardiovascular and diabetes risk factors when using protease inhibitors.

Los inhibidores de la proteasa (IP), que forman parte de la terapia HAART (terapia antirretroviral altamente activa), son inhibidores selectivos y competitivos de la proteasa, enzima crucial para la maduración, infección y replicación viral. Un síndrome lipodistrófico, asociado a hiperglucemia, hipercolesterolemia, hipertrigliceridemia, hiperlipidemia, hipertensión e hiperinsulinemia, ha sido relatado en pacientes tratados con HAART. Las anomalías metabólicas asociadas a la HAART fueron relacionadas, inicialmente, a los inhibidores de la proteasa, principalmente el Ritonavir, pero los mecanismos que relacionados a estas alteraciones metabólicas son poco comprendidos. Dado que los IP son posibles candidatos a fármacos de elección para tratamiento de otras enfermedades, como hepatitis C, malaria y algunos tipos de cáncer, es importante esclarecer las alteraciones metabólicas asociadas a los inhibidores de la proteasa. Ratas Wistar fueron tratadas dos veces por semana con 30 mg/kg de Ritonavir por 4 y 8 semanas. Fueron determinados los niveles de colesterol total, HDL, LDL, VLDL, triglicéridos y glucemia, al final de cada período considerado. Para evitar la interferencia de la ingestión de alimentos en las determinaciones de laboratorio, los animales fueron sometidos a un ayuno previo de 16 horas. Nuestros resultados mostraron un rápido aumento sérico de los niveles de triglicéridos, colesterol total, LDL-C y glucemia. No se observaron diferencias significativas para los niveles séricos de HDL-C o VLDL. Nuestro estudio apunta a la importancia de considerar los posibles antecedentes familiares de dislipidemia o diabetes, y controlar cualquier otro factor de riesgo cardiovascular y de diabetes cuando se utilizan los inhibidores de la proteasa.

Composição química e digestibilidade ruminal in situ da forragem de quatro espécies do gênero Brachiaria / Chemical composition and in situ ruminal degradability of four Brachiaria species

Avaliaram-se a composição química e a cinética da digestibilidade ruminal in situ da matéria seca (MS), proteína bruta (PB) e fibra em detergente neutro (FDN) da forragem de quatro espécies de Brachiaria, com 56 dias de crescimento, coletadas por corte manual. Foram utilizadas três vacas mestiças Holandês x Zebu, fistuladas no rúmen. Foram observados teores de 21,0; 21,1; 20,5 e 19,6 por cento de MS; 7,5; 6,4; 6,8 e 7,0 por cento de PB; e 66,8; 70,1; 73,4 e 63,9 por cento de FDN, respectivamente, para B. brizantha, B. decumbens, B. humidicola e B. ruziziensis. O maior valor nutricional foi da B. ruziziensis, que apresentou as mais elevadas taxas de degradação e degradabilidades efetivas (DE) da MS e da PB, e o menor teor de FDN associado à maior taxa de degradação desse nutriente. A forragem de pior qualidade nutricional foi B. humidicola, com menores valores de DE da MS e PB e maiores concentrações das frações fibrosas e indigestíveis.

The chemical composition and the ruminal in situ degradability of dry matter (DM), crude protein (CP), and neutral detergent fiber (NDF) of four species of Brachiaria were evaluated. The forages were harvested at 56 days of growth. Three rumen-fistulated crossbred Holstein × Zebu cows were used. The chemical composition of B. brizantha, B. decumbens, B. ruziziensis, and B. humidicola forages were, respectively: 21.0, 21.1, 20.5, and 19.6 percent for DM; 7.5, 6.4, 6.8, and 7.0 percent for CP; and 66.8, 70.1, 73.4, and 63.9 percent for NDF. B. ruziziensis presented the best nutritional value among the Brachiaria species, as indicated by the highest effective degradability (ED) and ruminal degradation rates of both DM and CP. B. ruziziensis also showed the highest NDF ruminal degradation rate and the lowest NDF concentration. In contrast, B. humidicola showed the worst nutritional composition, as indicated by the lowest DM and CP ED values and the highest content of indigestible and fibrous fractions.

Regional differences in diffusion tensor imaging measurements: assessment of intrarater and interrater variability.

BACKGROUND AND PURPOSE: Diffusion tensor imaging (DTI) has become a valuable tool in both the research and clinical evaluation of subjects. We sought to quantify interobserver and intraobserver variability of diffusivity and diffusion anisotropy measurements with regard to specific regions of interest (ROIs). MATERIALS AND METHODS: The subject group consisted of 5 healthy control subjects and 7 study subjects (all males; 16-19 years old; mean age = 17.5 years), as part of a protocol for closed head injury. Two whole-brain DTI scans were acquired on a 3T scanner for each subject. Analysis was performed using a ROI approach. Two independent observers analyzed the apparent diffusion coefficient (ADC) and fractional anisotropy (FA) indices in the corpus callosum, cortical spinal tract, internal capsules (ICs), basal ganglia, and centrum semiovale (CSO). Intraobserver and interobserver variability were calculated for the mean ADC, FA, and ordered eigenvalues of the diffusion tensor (lambda(1), lambda(2), and lambda(3)). RESULTS: The overall kappa statistic for intraobserver variability for both observers showed slight-to-substantial agreement (kappa = 0.02-0.69), however FA values in the CSO showed only slight agreement. Interobserver agreement was also slight to substantial for these DTI measurements with high variability in FA values in the IC and CSO. CONCLUSIONS: When one is comparing 2 DTI measurements, it is important to assess intraobserver and interobserver variability. We recommend caution in the analysis of DTI contrasts in the IC and CSO, because we have found the widest range of variability in measurements within these structures.

Intraosseous lipoma of the left frontoethmoidal sinuses and nasal cavity.

Intraosseous lipomas are very rare slow-growing benign tumors that may appear as congenital lesions or may be acquired (metaplasia from a pre-existing lipoma). Only a handful of head and neck cases have been reported in the literature. We present the first reported case of a solitary osteolipoma involving the sinonasal tract in a 66-year-old man with sinonasal symptoms. CT showed a lesion involving the left frontal sinus, extending into the ethmoid sinus with insinuation along the left middle turbinate. It appeared to be an atypical mass with areas of high attenuation (calcification) within.

Oral leishmaniasis: a clinicopathological study of 11 cases.

Leishmaniasis is a parasitic disease with diverse clinical manifestations, and considered a public health problem in endemic countries such as Brazil. Mucosal lesions usually involve the upper respiratory tract, with a predilection for nose and larynx. Oral involvement is unusual and in most cases it becomes evident after several years of resolution of the original cutaneous lesions. Oral lesions classically appear as mucosal ulcerations, mainly in the hard or soft palate. This report describes the clinicopathological data of 11 cases of mucocutaneous leishmaniasis with oral manifestations. Two cases of Leishmania (Viannia) braziliensis and one case of Leishmania (Leishmania) amazonensis were confirmed by polymerase chain reaction-restriction fragment length polymorphism or DNA sequencing in mucosal samples.

Modifications of Phleum pratense grass pollen allergens following artificial exposure to gaseous air pollutants (O(3), NO(2), SO(2)).

BACKGROUND: Air pollution is frequently proposed as a potential cause of the increased incidence of allergy in industrialised countries. Our objective was to investigate the impact of the major gaseous air pollutants on grass pollen allergens. METHODS: Timothy grass pollen was exposed to ozone (O(3)), nitrogen dioxide (NO(2)) and sulphur dioxide (SO(2)) alone or in combination. Allergen contents were analysed by 2-dimensional immunoblot using grass pollen-sensitive patient sera. RESULTS: For O(3)-treated pollen, immunoblotting showed an acidification of allergens Phl p 1b, Phl p 4, Phl p 5 and Phl p 6 and an IgE recognition decrease in Phl p 1, Phl p 2, Phl p 6 and Phl p 13. NO(2) exposure induced a decrease in Phl p 2, Phl p 5b and Phl p 6 recognition, and SO(2) treatment induced a decrease in Phl p 2, Phl p 6 and Phl p 13 recognition. Moreover, samples treated with a mix of NO(2)/O(3) or NO(2)/SO(2) showed a higher decrease in allergen content, compared with samples treated with only one pollutant. The O(3) acidification was also observed with the NO(2)/O(3) mix. CONCLUSION: Exposure of pollen to gaseous pollutants induced a decrease in allergen detection in pollen extracts. This decrease could be due to a mechanical loss of allergens from the altered pollen grains and/or post-translational modifications affecting allergen recognition by IgE.

Imaging findings of a hibernoma of the neck.

Hibernoma is an uncommon benign fatty tumor that arises from the vestiges of fetal brown fat. We present a case report of a hibernoma of the neck in an asymptomatic 19-year-old girl and describe the important imaging findings. Computed tomography (CT) shows a well defined hypodense mass with septations. Magnetic resonance imaging (MRI) shows intermediate T1 and bright T2 signal of the mass and also demonstrates the characteristic marked contrast enhancement.

Traffic-related air pollutants induce the release of allergen-containing cytoplasmic granules from grass pollen.

BACKGROUND/AIM: Pollen cytoplasmic granules (PCG) are loaded with allergens. They are released from grass pollen grains following contact with water and can form a respirable allergenic aerosol. On the other hand, the traffic-related air pollutants NO2 and O3 are known to be involved in the current increase in the prevalence of allergic diseases via their adjuvant effects. Our objective was to determine the effects of air pollutants on the release of PCG from Phleum pratense (timothy grass) pollen. METHODS: P. pratense pollen was exposed to several concentrations of NO2 and O3. The induced morphological damages were observed by environmental scanning electron microscopy, and the amount of PCG released from the pollen upon contact with water was measured. RESULTS: The percentages of damaged grain were 6.4% in air-treated controls, 15% after treatment with the highest NO2 dose (50 ppm) and 13.5% after exposure to 0.5 ppm O3. In treated samples, a fraction of the grains spontaneously released their PCG. Upon subsequent contact with water, the remaining intact grains released more PCG than pollen exposed to air only. CONCLUSIONS: Traffic-related pollutants can trigger the release of allergen-containing granules from grass pollen, and increase the bioavailability of airborne pollen allergens. This is a new mechanism by which air pollution concurs with the current increase in the prevalence of allergic diseases.

Intoxicação por enrofloxacina em um cão da raça Pinscher Miniatura: relato de caso / Enrofloxacin poisoning in a Miniature Pinscher: a case report

Uma cadela da raça Pinscher Miniatura foi medicada pelo proprietário com enrofloxacina, na dose de 50mg/kg, uma vez ao dia, por dois dias (dose diária 10 vezes maior que a prescrita). Ao exame clínico o animal apresentou-se deprimido, em cifose lombar, hipotérmico, com mucosas pálidas, dispnéia, sialorréia, vômitos e anúria, evoluindo para parada respiratória, convulsões e coma. Foi realizado tratamento sintomático. O fluxo urinário retornou ao normal em 12 horas e as convulsões foram controladas, mas o animal permaneceu em coma, morrendo 72 horas após o início do tratamento. A necropsia e o histopatológico confirmaram insuficiência renal e hepática agudas, e choque hipovolêmico, compatível com intoxicação por enrofloxacina.

A Miniature Pinscher bitch was treated by the owner with enrofloxacin at dose of 50 mg/kg, once a day, for two days (daily dose rate 10 times greater than prescribed). Physical examination showed depression, lumbar cifosis, hipotermia, pale mucosa, dispneia, drewling, vomiting and anuria, followed by respiratory failure, seizures, and coma. Symptomatic treatment was performed, and the urinary flow returned to normal in 12 hours, and seizures were controlled. However, the animal stayed in coma, and died 72 hours after the beginning of the treatment. Acute renal and hepatic failure, and hipovolemic shock, compatible with enrofloxacin intoxication, were observed through necropsia and histopatology.

Intratracheal instillation of cytoplasmic granules from Phleum pratense pollen induces IgE- and cell-mediated responses in the Brown Norway rat.

BACKGROUND: Release of cytoplasmic granules from grass pollen upon contact with water is thought to be an important source of airborne allergens. OBJECTIVES: To investigate the humoral and cellular responses to intratracheal instillation of Phleum pratense (timothy grass) pollen cytoplasmic granules (PCG) in the Brown Norway rat. METHODS: PCG were purified from timothy grass pollen by filtration through 5-microm-mesh filters. Rats were sensitized (day 0) and challenged (day 21) intratracheally with purified PCG suspended in saline (6 x 10(6) PCG/rat). Rats were then challenged 4 weeks later (1.5 x 10(6) PCG/rat). Blood samples, bronchial lymph nodes and lungs were collected from the rats 4 days after the second challenge. PCG-specific IgE and IgG1 levels and specificity were determined by ELISA and Western blotting. Pollen, pollen extract and PCG-induced proliferation of lymph node cells were monitored by [(3)H]-thymidine incorporation in a lymph node assay. Histopathological examination was carried out on the lungs. RESULTS: Specific IgE and IgG1 were present in the sera. Cultured lymph node cells proliferated in the presence of pollen, pollen extract and PCG. Western blots showed that all major pollen allergens are recognized by IgE and IgG1 from PCG-treated rats. Histopathological examination revealed features of a mild allergic reaction. CONCLUSIONS: In our rat model of allergy, purified timothy grass PCG instillation induced specific antibodies and lymph node cell responses, comparable to those obtained with intact pollen.

Prevalence assessment of periodontal disease in 3-6 year old children through PSR--a pilot study.

Increasing amounts of information emphasise the relevance of prevention, early diagnosis and early treatment of periodontal diseases in children. Children and adolescents affected by periodontal disease, mainly those who present a fast and severe attachment loss, are considered to be at risk of developing early or advanced periodontitis. Alternatively they may be presenting a reflex of systemic conditions affecting the periodontium. This study was aimed at verifying the acceptability of the use of the Simplified PSR (Periodontal Screening and Recording) Index in a very young population since a previous study in Bahia, Brazil, indicated a very high need for periodontal treatment in adolescents and young adults. A total of 200 children aged 3-6 years from private schools in Bahia, Brazil, were examined by four trained undergraduate students. The screening system was well accepted by the subjects and the fact that it is a fast tool was considered important for the successful examination of all sextants without behavioural disturbance. A high prevalence and a low severity of parameters related to periodontal disease were found in this population. A statistically higher prevalence of PSR code 2 (61.5%) [54.50-68.49] CI 95% when compared to PSR codes 0 (23.5%) 117.93-30.10] CI 95%, 1 (14.5%) 110.07-20.32] CI 95% and 3 (0.5%) 10.02-3.18] CI 95% were shown. There was no statistically significant difference between female and male children for any PSR code. The finding of more parameters related to periodontal health in S5 when compared to sextants S6 and S4 showed statistical significance (CI 95%).

Periodontal disease associated with Langerhans' cell histiocytosis: case report.

A clinical case of Langerhans' cell histiocytosis, type eosinophilic granuloma, in a young adult patient is presented. Because of the occurrence of oral manifestations in initial stages of the disease, there is a need for a differential diagnosis, especially with the early-onset periodontitis.