Revealing the microbial diversity and physicochemical characteristics of Brazilian untreated green table olives.

AIM: This work evaluated the microbial diversity and physicochemical characteristics of fresh and fermented fruits from Brazilian untreated green table olives of the Ascolano and Grappolo cultivars. METHODS AND RESULTS: Twenty species of mesophilic bacteria, seven lactic acid bacteria, and fourteen yeast were identified. Some species prevailed over others, such as the bacteria Levilactobacillus brevis, Lacticaseibacillus paracasei subsp. paracasei, Pantoea agglomerans, Staphylococcus warneri, Bacillus simplex, B. thuringiensis, and the yeasts Candida parapsilosis, Ca. orthopsilosis, and Cryptococcus flavescen. In the olive fruit and olive brine, the sugars: sucrose, glucose, mannitol, and fructose, and the acids: acetic, citric, lactic, malic, and succinic were identified. Thirty-seven volatile compounds belonging to different chemical classes of acids, alcohols, aldehydes, esters, hydrocarbons, phenols, ketones, and ether were identified in the fruits and brine olives. CONCLUSION: The polyphasic methodology using matrix assisted laser desorption/ionization-time of flight and 16S rRNA sequencing was efficiently performed to identify microorganisms; chemical analysis helped to understand the fermentation process of olives.

Cellular and Biochemical Effects of Combined X-Ray Radiation and Storage on Whole Blood.

BACKGROUND: Evaluating the impact of ionizing radiation on stored blood is relevant since blood banks are major assets in emergency conditions such as radiation incident/attack. This study aimed to fill our knowledge gap of combined radiation and storage effects on blood. METHODS: Blood collected from 16 anesthetized rats was anticoagulated, aliquoted into storage bags, and assigned to 8 groups using protocols combining storage (1-day vs 3-day 4oC) plus irradiation (75 Gy vs 0 Gy - control). Bags were positioned inside an X-ray irradiator (MultiRad-350). Complete blood count, differential white blood cell count, biochemistry, and hemostasis were analyzed (≥7 bags/group). RESULTS: Na+, bicarbonate, glucose, and pH significantly reduced, while K+, Cl-, and lactate increased by storage. Coagulation measures were not significantly altered after radiation. White blood cell count and most cell types were numerically reduced after radiation, but changes were statistically significant only for monocytes. No significant alterations were noted in aggregation or rotational thromboelastometry parameters between irradiated and control. CONCLUSIONS: Evaluating cellular/biochemical parameters aids in assessing stored blood adequacy after radiation. Data suggest that fresh or cold-stored blood can sustain up to 75 Gy without major critical parameter changes and may remain suitable for use in critically ill patients in military/civilian settings.

Copper Toxicity in Horses: Does it Exist?

Copper toxicity is thought to be a rare condition in horses. However, the number of cases diagnosed in Brazil is growing. This article aims to describe cases of copper toxicity involving horses from different geographic locations and discuss findings of physical examinations, differential diagnoses and potential causes. Five cases referred from 4 different properties where at least 15 other horses were affected were described. Hemolytic anemia and hemoglobinuria, presence of Heinz bodies and elevated aspartate aminotransferase and gamaglutamil transferase levels were detected in all cases. The diagnosis was based on clinical history and signs, laboratory tests results, copper level determination in feed and/or soil and histopathological findings. Two horses progressed to acute death; remaining horses responded to clinical management with or without blood transfusion, depending on disease severity. However, one of these horses, after several returns to the veterinary hospital, was euthanized due to complications. One horse was treated with ammonium tetrathiomolybdate. Two horses had several recurring episodes over the course of several months, an uncommon presentation in ruminants suffering from copper toxicity. Excess copper was associated with soil fertilization with poultry litter or treatment of previous or neighbor crops with copper-containing products. It can be concluded that copper toxicity does occur in horses and may arise from several sources and/or be associated with predisposing dietary factors. Given the growing number of cases, the condition should be included in the differential diagnosis list and proper preventive dietary and pasture fertilization measures adopted.

Role of albumin on endothelial basement membrane and hemostasis in a rat model of hemorrhagic shock.

BACKGROUND: We sought to determine the extent of loss of endothelial basement membrane (BM), leukocyte recruitment, and changes in coagulation after hemorrhagic shock, followed by limited-volume resuscitation (LVR) with 5% albumin (ALB). METHODS: Anesthetized rats were bled 40% of blood volume and assigned to treatment groups: untreated (n = 6), LVR with normal saline (NS; n = 8), or LVR with ALB (n = 8). Sham rats (n = 6) underwent all procedures except hemorrhage or resuscitation. Blood samples were assayed for active proteases, such as metalloproteinase 9 (MMP-9) and a disintegrin and metalloproteinase 10 (ADAM-10), BM-type heparan sulfate proteoglycan (perlecan), cell count, and coagulation function. Leukocyte transmigration was used to estimate the net efficiency of leukocyte recruitment in cremaster venules. RESULTS: Hemorrhage significantly lowered red cell count, but white cell and platelet counts did not change (vs. sham). Ionized calcium in plasma was significantly reduced in untreated and remained so after NS. In contrast, ionized calcium was normalized after ALB. Plasma expansion after NS and ALB further reduced leukocyte and platelet counts. Metalloproteinase 9, ADAM-10, and perlecan were significantly higher in untreated rats (vs. sham). Albumin normalized MMP-9, ADAM-10, and perlecan levels, while NS further increased MMP-9, ADAM-10, and perlecan (vs. sham). Transmigrated leukocytes doubled in the untreated group and remained elevated after NS (vs. sham) but normalized after ALB. Albumin reduced every stage of the leukocyte recruitment process to sham levels. CONCLUSION: Despite similar plasma expansion, NS weakened platelet function contrary to ALB. Plasma expansion with ALB resulted in restoration of BM integrity and attenuation of leukocyte recruitment to tissues, in contrast to NS. Albumin plays a critical role in restoring BM integrity, attenuating leukocyte recruitment to tissues, and optimizing hemostasis by increasing ionized calcium in plasma.

Detection and Molecular Characterization of a Natural Coinfection of Marek's Disease Virus and Reticuloendotheliosis Virus in Brazilian Backyard Chicken Flock.

Marek's disease virus (MDV) and the reticuloendotheliosis virus (REV) are two of the primary oncogenic viruses that significantly affect chickens. In Brazil, there have been no previous published reports on the presence of field REV alone or in coinfection. This retrospective study analyzes samples from a case of lymphoproliferative lesions from a backyard chicken flock. MDV and REV were detected by PCR and classified as MDV1 and REV3, respectively, through sequencing and phylogenetic analysis based on the glycoprotein B (gB) genes for MDV and the polymerase (pol) and envelope (env) genes for REV. Real-time PCR reactions were performed for MDV to rule out the presence of the Rispens vaccine strain. This is the first report of the presence of REV in coinfection with a MDV clinical case in Brazil and the first molecular characterization of REV in South America. This study highlights the importance of molecular diagnosis for REV and MDV in poultry. In addition, this study highlights the distribution of these two viruses worldwide and the latent risk of them solely or in coinfection to this part of the world.

Clinical, histopathological and immunohistochemical chacteristics of equine cutaneous mastocytoma: case report / Características clínicas, histopatológicas e imuno-histoquímicas do mastocitoma cutâneo equino: relato de caso

A 3 year-old mare, Arab, that belongs to the experimental herd of FMVZ-USP presented cutaneous tumor lesions on right and left posterior limb with approximately one year of evolution. A new formation with rapid growth was observed on the neck region, and since then, the limbs lesion gained inflammatory characteristics. Aspirative punction was performed on the neck formation in order to submit the sample to cytological analyses, in which eosinophilic infiltrate was identified. At this point, a conservative approach was chosen with three consecutive corticoid infiltrations. With lack of success of infiltrative therapy, neck, right and left posterior limbs lesions were surgically removed. The material obtained from surgical procedures were sent to histophatological and immunohistrochemical evaluation, that confirmed cutaneous mastocytoma diagnosis. The mare was discharged after all surgical wound were healed and after full remission of clinical signs, and did not show relapses or any other complication from tumor lesions. Some cutaneous tumors are well described and commonly occur in horses, such as sarcoids, melanoma, papilloma and squamous cell carcinoma. Cutaneous mastocytoma is considered a rare cutaneous tumor in this species. According to literature, equine mastocytoma do not have breed predilection, however, some authors mention a greater involvement in Purebreed Arabian horses. In order to establish a definitive diagnosis, it is important to associate physical exams to histophatological an immunohistochemical evaluation and, even though rare in horses, this neoplasm must be considered as a differential diagnosis.

Uma égua puro sangue árabe, com 3 anos de idade, pertencente ao rebanho experimental da FMVZ-USP apresentou lesões tumorais cutâneas nos membros pélvicos direito e esquerdo com aproximadamente um ano de evolução. Notou-se nova formação cutânea com crescimento rápido na região do pescoço, e desde então, as lesões dos membros ganharam características inflamatórias. Foi realizada punção aspirativa da lesão do pescoço para análise citológica, em que se identificou infiltrado eosinofílico. Optou-se pelo início de tratamento conservativo com três infiltrações intralesionais consecutivas com corticoesteroide. A partir da falta de sucesso das infiltrações foram realizadas as excisões cirúrgicas totais dos nódulos do pescoço, membro pélvico direito e esquerdo. Todos os materiais obtidos dos procedimentos cirúrgicos foram enviados para avaliação histopatológica e imuno-histoquímica, nas quais se confirmou o diagnóstico de mastocitoma cutâneo. O animal recebeu alta após a cicatrização das feridas cirúrgicas e remissão dos sinais, e não demonstrou recidivas ou outras complicações advindas das lesões tumorais. Algumas neoplasias cutâneas são bem descritas e de comum ocorrência na espécie equina, como o sarcoide, melanoma, papiloma e tumor de células escamosas. Já o mastocitoma cutâneo consiste em neoplasia cutânea rara nessa espécie. Segundo a literatura, não parece existir uma predileção racial para o aparecimento deste tumor, entretanto, alguns autores citam o acometimento maior nos animais Puro Sangue Árabe. Para o estabelecimento do diagnóstico definitivo é importante a associação dos exames físico, histopatológico e imuno-histoquímico e, apesar desta neoplasia ser de raro aparecimento nos equinos, deve ser sempre considerada como diagnóstico diferencial.

Características clínicas, histopatológicas e imuno-histoquímicas do mastocitoma cutâneo equino: relato de caso / Clinical, histopathological and immunohistochemical chacteristics ofequine cutaneous mastocytoma: case report

Uma égua puro sangue árabe, com 3 anos de idade, pertencente ao rebanho experimental da FMVZ-USP apresentou lesões tumorais cutâneas nos membros pélvicos direito e esquerdo com aproximadamente um ano de evolução. Notou-se nova formação cutânea com crescimento rápido na região do pescoço, e desde então, as lesões dos membros ganharam características inflamatórias. Foi realizada punção aspirativa da lesão do pescoço para análise citológica, em que se identificou infiltrado eosinofílico. Optou-se pelo início de tratamento conservativo com três infiltrações intralesionais consecutivas com corticoesteroide. A partir da falta de sucesso das infiltrações foram realizadas as excisões cirúrgicas totais dos nódulos do pescoço, membro pélvico direito e esquerdo. Todos os materiais obtidos dos procedimentos cirúrgicos foram enviados para avaliação histopatológica e imuno-histoquímica, nas quais se confirmou o diagnóstico de mastocitoma cutâneo. O animal recebeu alta após a cicatrização das feridas cirúrgicas e remissão dos sinais, e não demonstrou recidivas ou outras complicações advindas das lesões tumorais. Algumas neoplasias cutâneas são bem descritas e de comum ocorrência na espécie equina, como o sarcoide, melanoma, papiloma e tumor de células escamosas. Já o mastocitoma cutâneo consiste em neoplasia cutânea rara nessa espécie. Segundo a literatura, não parece existir uma predileção racial para o aparecimento deste tumor, entretanto, alguns autores citam o acometimento maior nos animais Puro Sangue Árabe. Para o estabelecimento do diagnóstico definitivo é importante a associação dos exames físico, histopatológico e imuno-histoquímico e, apesar desta neoplasia ser de raro aparecimento nos equinos, deve ser sempre considerada como diagnóstico diferencial.

A 3 year-old mare, Arab, that belongs to the experimental herd of FMVZ-USP presented cutaneous tumor lesions on right and left posterior limb with approximately one year of evolution. A new formation with rapid growth was observed on the neck region, and since then, the limbs lesion gained inflammatory characteristics. Aspirative punction was performed on the neck formation in order to submit the sample to cytological analyses, in which eosinophilic infiltrate was identified. At this point, a conservative approach was chosen with three consecutive corticoid infiltrations. With lack of success of infiltrative therapy, neck, right and left posterior limbs lesions were surgically removed. The materials obtained from surgical procedures were sent to histophatological and immunohistrochemical evaluation that confirmed cutaneous mastocytoma diagnosis. The mare was discharged after all surgical wound were healed and after full remission of clinical signs, and did not show relapses or any other complication from tumor lesions. Some cutaneous tumors are well described and commonly occur in horses, such as sarcoids, melanoma, papilloma and squamous cell carcinoma. Cutaneous mastocytoma is considered a rare cutaneous tumor in this species. According to literature, equine mastocytoma do not have breed predilection, however, some authors mention a greater involvement in Purebreed Arabian horses. In order to establish a definitive diagnosis, it is important to associate physical exams to histophatological an immunohistochemical evaluation and, even though rare in horses, this neoplasm must be considered as a differential diagnosis.

Maduración pulmonar fetal: efectos sobre la morbilidad respiratoria en prematuros tardíos / Fetal pulmonary maturation: effects on respiratory morbidity in late preterm infants

INTRODUCCIÓN: Los recién nacidos entre las 34 y las 36 semanas de gestación (SEG), definidos actualmente como "prematuros tardíos" (RNPTT) requieren en mayor frecuencia de internación en unidades de cuidados intensivos neonatales, siendo la morbilidad respiratoria una de las principales causas de internación. El uso de corticoides prenatales ha demostrado mejorar la función pulmonar en prematuros extremos (<27,6 SEG), muy prematuros (28-31,6 SEG) y prematuros moderados, (32-36,6 SEG) pero no han sido evaluados en forma extensa en el período prematuro tardío y no existen datos concluyentes al respecto. OBJETIVO: Comparar la frecuencia en que se presenta la morbilidad respiratoria en RNPTT con o sin maduración pulmonar previa. Establecer los factores asociados a morbilidad respiratoria en RNPTT. Comparar los pacientes RNPTT con y sin maduración pulmonar: los requerimientos de internación en general, requerimiento de internación por morbilidad respiratoria y en estos, días de oxigenoterapia y necesidad de ARM y/o VNI. MÉTODO: Diseño observacional, retrospectivo y analítico. Población constituida por todos los RNPTT que nacieron en la CURF entre el 1 de enero de 2017 y el 31 de diciembre de 2017. Las variables maternas y neonatales se compararán entre los dos grupos RNPTT con o sin maduración pulmonar. La frecuencia de presentación de la morbilidad respiratoria entre los RNPTT con o sin maduración pulmonar, los factores asociados a morbilidad respiratoria, los requerimientos de internación y necesidad de ARM y VNI se compararon con la prueba de chicuadrado siendo estadísticamente significativa la diferencia entre los mismos con P ≤0,05. Los días de internación se compararon mediante la prueba de T de Student. El Software que se utilizó para procesar los datos estadísticos será el SPSS versión 11.0 (Chicago, IL). RESULTADOS: De los RNPTT estudiados (160) N=86 (53.7 %) recibieron corticoides prenatales completos. La edad gestacional media (DE) de administración fueron 35,2 (2,2) semanas. La frecuencia de morbilidad respiratoria fue de 21,2%. No hubo diferencia en la frecuencia de morbilidad respiratoria global entre ambos grupos (p 0,3777). El sexo masculino (51,3% vs 49.0%; p= 0,0034) la APP (34,09%; p= 0,0197) y morbilidad materna (preeclampsia, app, diabetes gestacional, corioamnionitis, RPM, placenta previa, HIE) (70,65%; p= 0,0062) se asociaron a morbilidad respiratoria. No hubo diferencias en el requerimiento de internación ni en el número de días de internación entre ambos grupos. Tampoco se observó diferencias en el requerimiento de oxígeno, ARM, VNI, uso de surfactante y necesidad de reanimación. Tampoco se hallaron diferencias en la frecuencia de enfermedad de membrana hialina, taquipnea transitoria del recién nacido, neumotórax. Hubo tendencia a una disminución de la frecuencia de SDRA (6,35 % vs 33.33%, p= 0,051) en el grupo que recibió maduración pulmonar como así también una mayor frecuencia de apneas 25% (p= 0,0392). CONCLUSIÓN: En este estudio la maduración pulmonar con corticoides no influencio la frecuencia de morbilidad respiratoria global. Sin embargo, al analizar las patologías respiratorias de manera individual, encontramos una tendencia a la disminución de SDRA en el grupo madurado. La frecuencia de apneas fue mayor en el grupo que recibió corticoides lo que requiere un análisis con mayor profundidad. El sexo masculino como así también la APP y morbilidad materna se asociaron a mayor frecuencia de morbilidad respiratoria. (AU)

INTRODUCTION: Newborns between 34 and 36 weeks of gestation (WG), currently defined as "latepreterm infants" (LPTI), require of admission in neonatal intensive care units more frequently, with respiratory morbidity being one of the main causes of hospitalization. The use of prenatal corticosteroids has been shown to improve lung function in extremely preterm (<27.6 WG), very preterm (28-31.6 WG), and moderate preterm infants (32-36.6 WG), but it has not been evaluated extensively in the late-preterm period, leaving no conclusive data in this regard. OBJECTIVES: To compare the frequency of respiratory morbidity in LPTI with and without previous lung maturation. To establish the factors associated with respiratory morbidity in LPTI. • To compare LPTI patients with and without lung maturation: the requirements for hospitalization in general, the need for hospitalization due to respiratory morbidity, and in these cases, days of oxygen therapy and need for MRA and / or NIV. METHOD: Observational, retrospective and analytical design. Population constituted by all LPTI that were born in the CURF between January 1, 2017 and December 31, 2017. Maternal and neonatal variables were compared between LPTI groups with and without lung maturation. The frequency of presentation of respiratory morbidity among LPTI with or without lung maturation, the factors associated with respiratory morbidity, the requirements for hospitalization and the need for MRA and NIV were compared with the chi-square test, considering statistically significant the difference between them with P ≤0.05. Days of hospitalization were compared using the Student's T test. The software used to process the statistical data was the SPSS version 11.0 (Chicago, IL). RESULTS: Of the LPTI studied (160), N = 86 (53.7%) received complete antenatal corticosteroids. Mean gestational age (SD) for administration was 35.2 (2.2) weeks. The frequency of respiratory morbidity was 21.2%. There was no difference in the frequency of overall respiratory morbidity between both groups (p 0.3777). Male sex (51,3% vs 49.0%; p= 0,0034), preterm birth risk (34,09%; p= 0,0197) and maternal morbidity (preeclampsia, preterm birth risk, gestational diabetes, chorioamnionitis, preterm membrane rupture, previous placenta, pregnancy-induced hypertension) (70,65%; p= 0,0062) were associated to respiratory morbidity. There were no differences regarding hospitalization requirement or the number of hospitalization days between both groups. No differences were observed in oxygen requirement, MRA, NIV, use of surfactant and need for resuscitation. No differences were found in the frequency of hyaline membrane disease, newborn transient tachypnea and pneumothorax. There was a tendency to a decrease in the frequency of RDS (6.25% vs 33.33%; p= 0.051), and a higher frequency of apneas (25.0% vs 0%; p= 0.0392) in the group of patients who received lung maturation. CONCLUSION: In this study, pulmonary maturation with corticosteroids did not influence the frequency of global respiratory morbidity. However, when analyzing respiratory pathologies individually, we found a tendency to decrease of RDS in the matured group. The frequency of apneas was higher in the group that received corticosteroids, which requires further analysis. Male sex as well as preterm birth risk and maternal morbidity were associated to a higher frequency of respiratory morbidity

Putative progressive and abortive feline leukemia virus infection outcomes in captive jaguarundis (Puma yagouaroundi).

BACKGROUND: Feline leukemia virus (FeLV) is an exogenous gammaretrovirus of domestic cats (Felis catus) and some wild felids. The outcomes of FeLV infection in domestic cats vary according to host susceptibility, virus strain, and infectious challenge dose. Jaguarundis (Puma yagouaroundi) are small wild felids from South and Central America. We previously reported on FeLV infections in jaguarundis. We hypothesized here that the outcomes of FeLV infection in P. yagouaroundi mimic those observed in domestic cats. The aim of this study was to investigate the population of jaguarundis at Fundação Parque Zoológico de São Paulo for natural FeLV infection and resulting outcomes. METHODS: We investigated the jaguarundis using serological and molecular methods and monitored them for FeLV-related diseases for 5 years. We retrieved relevant biological and clinical information for the entire population of 23 jaguarundis held at zoo. Post-mortem findings from necropsies were recorded and histopathological and immunohistopathological analyses were performed. Sequencing and phylogenetic analyses were performed for FeLV-positive samples. For sample prevalence, 95% confidence intervals (CI) were calculated. Fisher's exact test was used to compare frequencies between infected and uninfected animals. P-values <0.05 were considered significant. RESULTS: In total, we detected evidence of FeLV exposure in four out of 23 animals (17%; 95% CI 5-39%). No endogenous FeLV (enFeLV) sequences were detected. An intestinal B-cell lymphoma in one jaguarundi was not associated with FeLV. Two jaguarundis presented FeLV test results consistent with an abortive FeLV infection with seroconversion, and two other jaguarundis had results consistent with a progressive infection and potentially FeLV-associated clinical disorders and post-mortem changes. Phylogenetic analysis of env revealed the presence of FeLV-A, a common origin of the virus in both animals (100% identity) and the closest similarity to FeLV-FAIDS and FeLV-3281 (98.4% identity), originally isolated from cats in the USA. CONCLUSIONS: We found evidence of progressive and abortive FeLV infection outcomes in jaguarundis, and domestic cats were probably the source of infection in these jaguarundis.

Low-volume resuscitation with normal saline is associated with microvascular endothelial dysfunction after hemorrhage in rats, compared to colloids and balanced crystalloids.

BACKGROUND: Restoration of endothelial glycocalyx (EG) barrier may be an essential therapeutic target for successful resuscitation. The aim of this study was to compare in vivo the effects of resuscitation with normal saline (NS) to lactated Ringer's solution (LR), 5% albumin and fresh frozen plasma (FFP) on their ability to maintain EG and barrier function integrity, mitigate endothelial injury and inflammation, and restore vascular homeostasis after hemorrhagic shock. METHODS: Anesthetized rats (N = 36) were subjected to hemorrhagic shock (bled 40% of total blood volume), followed by resuscitation with 45 ml/kg NS or LR, or 15 ml/kg 5% albumin or FFP. Microhemodynamics, EG thickness, permeability, leukocyte rolling and adhesion were assessed in >180 vessels from cremaster muscle, as well as systemic measures. RESULTS: After hypotensive resuscitation, arterial pressure was 25% lower than baseline in all cohorts. Unlike FFP, resuscitation with crystalloids failed to restore EG thickness to baseline post shock and shedding of glycocalyx proteoglycan was significantly higher after NS. NS decreased blood flow and shear, and markedly increased permeability and leukocyte rolling/adhesion. In contrast, LR had lesser effects on increased permeability and leukocyte rolling. Albumin stabilized permeability and white blood cell (WBC) rolling/adhesion post shock, comparable to FFP. CONCLUSIONS: Resuscitation with NS failed to inhibit syndecan-1 shedding and to repair the EG, which led to loss of endothelial barrier function (edema), decline in tissue perfusion and pronounced leukocyte rolling and adhesion. Detrimental effects of NS on endothelial and microvascular stabilization post shock may provide a pathophysiological basis to understand and prevent morbidity associated with iatrogenic resuscitation after hemorrhagic shock.

Novel Adjunct Drugs Reverse Endothelial Glycocalyx Damage After Hemorrhagic Shock in Rats.

INTRODUCTION: There is interest in the small-volume therapeutic use of adjunct drugs for treating hemorrhagic shock (HS). However, critical information is only partially available on mechanisms of action of promising compounds such as adenosine-lidocaine-magnesium (ALM), beta-hydroxybutyrate plus melatonin (BHB/M), and poloxamer 188 (P-188). Therefore, we tested the hypothesis that these adjuncts would reverse HS-induced damage to microvascular endothelial glycocalyx and hemodynamics. METHODS: After baseline, 40% of total blood volume was removed from 44 anesthetized Sprague-Dawley male rats. One hour after hemorrhage, animals were resuscitated using ALM, BHB/M, or P-188 followed by lactated Ringer's (LR, 15 mL/kg). Control animals were not treated (SHAM) or received LR alone. Sampled blood was used to quantify shed syndecan-1 in plasma; multiple systemic physiological parameters were recorded. In vivo glycocalyx thickness, microvascular permeability, and microhemodynamics were evaluated in >200 cremaster venules using intravital videomicroscopy. RESULTS: Compared with baseline, resuscitation using adjuncts was associated with glycocalyx restoration of 97 ±â€Š9% (ALM), 75 ±â€Š8% (BHB/M), and 85 ±â€Š5% (P-188): significantly higher than LR-only (56 ±â€Š4%). Significantly better permeability, similar to SHAM values, was measured after ALM and P-188, and low plasma syndecan-1 levels were measured after resuscitation with all adjuncts. Microhemodynamic changes were relatively small while systemic parameters such as mean arterial pressure and lactate improved but remained below or above the baseline, respectively, as expected from this hypotensive resuscitation model. CONCLUSION: The drugs ALM, BHB/M, and P-188 provide beneficial effects as adjuncts to hypotensive resuscitation in this HS model by mechanisms involving changes at the microvascular level including the glycocalyx.

Cryptococcus gattii molecular type VGII infection associated with lung disease in a goat.

BACKGROUND: Cryptococcus gattii-induced cryptococcosis is an emerging infectious disease of humans and animals with worldwide distribution and public health importance due to its significant morbidity and mortality rate. The present study aimed to report a case of pulmonary infection by C. gattii molecular type VGII in State of São Paulo, Brazil. CASE PRESENTATION: A 5-year-old goat showing intermittent dry cough, ruminal tympany, anorexia, fever, tachycardia and tachypnea was presented for necropsy at the Veterinary Hospital of the School of Veterinary Medicine and Animal Sciences, São Paulo University, São Paulo, Brazil. Postmortem examination revealed numerous 2.0-6.0 cm diameter yellow gelatinous pulmonary masses. Tissues were evaluated by a combination of pathological, mycological, and molecular diagnostic techniques. Microscopically, pneumonia granulomatous, multifocal to coalescing, moderate, with many intralesional carminophilic yeasts was observed. The immunohistochemistry and mycological culture confirmed Cryptococcus spp. Internal transcribed spacers and orotidine monophosphate pyrophosphorylase nucleotide differentiation demonstrated that the isolate corresponds to the C. gattii VGII molecular subtype. CONCLUSIONS: To our knowledge, this is the first report of a pulmonary infection in a goat linked to C. gattii molecular type VGII in Southeastern Brazil. Our findings emphasize the need for an active surveillance program for human and animal new infections to improve the current public health policies due to expansion of the epidemiological niche of this important microorganism.

Plasma syndecan-1 and heparan sulfate correlate with microvascular glycocalyx degradation in hemorrhaged rats after different resuscitation fluids.

The endothelial glycocalyx plays an essential role in many physiological functions and is damaged after hemorrhage. Fluid resuscitation may further change the glycocalyx after an initial hemorrhage-induced degradation. Plasma levels of syndecan-1 and heparan sulfate have been used as indirect markers for glycocalyx degradation, but the extent to which these measures are representative of the events in the microcirculation is unknown. Using hemorrhage and a wide range of resuscitation fluids, we studied quantitatively the relationship between plasma biomarkers and changes in microvascular parameters, including glycocalyx thickness. Rats were bled 40% of total blood volume and resuscitated with seven different fluids (fresh whole blood, blood products, and crystalloids). Intravital microscopy was used to estimate glycocalyx thickness in >270 postcapillary venules from 58 cremaster preparations in 9 animal groups; other microvascular parameters were measured using noninvasive techniques. Systemic physiological parameters and blood chemistry were simultaneously collected. Changes in glycocalyx thickness were negatively correlated with changes in plasma levels of syndecan-1 (r = -0.937) and heparan sulfate (r = -0.864). Changes in microvascular permeability were positively correlated with changes in both plasma biomarkers (r = 0.8, P < 0.05). Syndecan-1 and heparan sulfate were also positively correlated (r = 0.7, P < 0.05). Except for diameter and permeability, changes in local microcirculatory parameters (red blood cell velocity, blood flow, and wall shear rate) did not correlate with plasma biomarkers or glycocalyx thickness changes. This work provides a quantitative framework supporting plasma syndecan-1 and heparan sulfate as valuable clinical biomarkers of glycocalyx shedding that may be useful in guiding resuscitation strategies following hemorrhage.

Tomography, Radiography, and Rhinoscopy in Diagnosis of Benign and Malignant Lesions Affecting the Nasal Cavity and Paranasal Sinuses in Dogs: Comparative Study.

Canine nasal and paranasal diseases have variable causes. Presumptive diagnosis is based on clinical manifestations; however, high similarity of clinical signs often calls for diagnostic imaging modalities and rhinoscopy before a definitive diagnosis can be reached. This study sets out to determine the value of rhinoscopy, radiography, and computed tomography (CT) of the head for canine nasal and paranasal disease diagnosis using a purposely developed comparative score. In all, 20 dogs presenting with clinical signs consistent with nasal disease were used. Patients were submitted to radiographic, CT, and rhinoscopic assessment; rhinoscopy-guided biopsy collection was performed in cases presenting with tissue proliferation, ulceration, or other nasal mucosal lesions. Rhinoscopy and rhinoscopy combined with CT significantly contributed to nasal disease diagnosis. Rhinoscopy and CT are complementary diagnostic modalities. Rhinoscopy proved helpful for confirmation of presumptive diagnosis and allowed image-assisted biopsy collection whereas CT contributed to effective determination of lesion extension and involvement of adjacent structures. Yet, histologic confirmation remains vital for definitive diagnosis.

Pneumonia and bacteremia in a golden-headed lion tamarin (Leontopithecus chrysomelas) caused by Klebsiella pneumoniae subsp. pneumoniae during a translocation program of free-ranging animals in Brazil.

Klebsiella pneumoniae is an important emerging pathogen in humans, particularly the invasive hypermucoviscosity (HMV) phenotype. In addition, the organism is an important public health concern because of nosocomial infections and antimicrobial resistance. Nonhuman primates in captivity are susceptible to Klebsiella, particularly when a stress factor is involved. Infections vary depending on the species but can cause significant morbidity and mortality in these animals. The objective of this study was to describe a case of bronchopneumonia and bacteremia caused by Klebsiella pneumoniae in a free-ranging golden-headed lion tamarin (Leontopithecus chrysomelas) caught and maintained in quarantine during a translocation program for conservation purposes. An adult male, that had showed emaciation and apathy, was clinically examined and, despite being provided supportive therapy, died 2 days after onset of clinical signs. At postmortem examination, generalized bilateral pneumonia and pericarditis were observed. Tissue samples were fixed in 10% formalin for histology, and pulmonary tissues and cardiac blood were collected for microbiologic diagnostic procedures. Bacteria that were shown to be HMV K. pneumoniae subsp. pneumoniae strains were isolated from the pulmonary fluids and cardiac blood in pure cultures. Severe bronchopneumonia was the main pathological finding. The consequences of the confirmed presence of the HMV phenotype of K. pneumoniae subsp. pneumoniae in this wildlife species for human, animal, and ecosystem health should be determined. These results demonstrate the importance of quarantine and potential pathogen screening during wildlife translocation procedures.

Effects of thermomechanical and thermocycling on the shear bond strength of brackets to bovine enamel bonded with conventional and self-etching adhesive systems / Efeito da ciclagem térmica e térmico-mecânica na resistência adesiva ao cisalhamento de bráquetes aderidos ao esmalte bovino com sistema adesivo convencional e auto-condicionante

Muitos sistemas adesivos foram desenvolvidos na última década. O objetivo desse estudo foi avaliara resistência adesiva ao cisalhamaneto (RAC)de bráquetes ortodônticos aderidos com primer convencional (PC) ou primer auto-condicionante (PAC), submetidos à ciclagem térmica (CT) e ciclagem térmico-mecânica (CTM). Incisivos bovinos (n =90)foram randomicamente divididos em 2 grupos (n =45) para a colagem dos bráquetes: XT (Transbond XT, 3M-Unitek, CA, USA); SEP (Transbond Plus SEP,3M-Unitek, CA, USA). Após o procedimento adesivo os grupos forma subdivididos em 3 sub-grupos: SC(sem ciclagem) – 1 e 4 (n =15 cada); CTM (ciclagem térmico-mecânica)- 2 e 5 (n = 15 cada); CT (ciclagem térmica)- 3 e 6 (n = 15 cada). Um simulador de fadiga térmica foi usado para os subgrupos 3 e 6, que fioram submetidos a 500 ciclos térmicos entre 5 and 55 C (ISO 11405). Os subgrupos 2 e 5 foram submetidos à ciclagem térmico-mecânica em um simulador de fadiga. Os espécimes foram colocados em uma basede metal num ângulo de 45 e os braquetes receberam impulsos de 1 bar e 4 Hz de frequência, por uma agulha de metal de 2.5 mm de diâmetro fixada na haste superior do equipamento. Foram submetidos a 100.000 ciclos mecânicos e 500 ciclos térmicos entre 5 and 55 C. O teste de RAC foi realizado emuma máquina universal de testes (1mm/min). Após orompimento da união foi realizada uma avaliação emmicroscopia óptica para avaliar a penetração adesivae o Índice de Permanência do Adesivo (IPA) sobreo esmalte. Os resultados mostraram maiores valores de RAC (14,70 ± 4,85 MPa) para o SEP. A CT não influenciou os resultados para o XT (NC- 11,44 MPa;TC - 11,20 MPa; TMC - 11,19 MPa), enquanto para oSEP, a CTM mostrou maiores valores (16,84 MPa)...

Several adhesive systems have been developed in the last decade. The aim of the current studywas to evaluate the shear bond strength (SBS) of orthodontic brackets bonded with conventional primers (XT) or self-etching primers (SEP), by means of thermocycling (TC) and thermomechanical(TM) procedures. 90 bovine incisors were randomly divided into 2 groups (45 specimens each) for brackets bonding: XT (Transbond XT, 3M-Unitek,CA, USA); SEP (Transbond Plus SEP, 3M-Unitek,CA, USA). After bonding, groups were divided into 3subgroups: NC (no cycling)- 1 and 4 (n = 15 each),TMC – 2 and 5 (n =15 each); and TC- 3 and 6 (n=15 each). The fatigue simulator was used for thethermal fatigue test of groups 3 and 6, which were submitted to thermal variation cycles - 500 cyles were carried out in between 5 and 55 C ( ISO 11405). Groups 2 and 5 were submitted to thermomechanicaltests by using the mechanical fatigue simulator. Thespecimens were placed at a metal base at an angleof 45, in a way that the 2.5 mm diameter metalneedle was fixed in the upper part of the cyclingmachine and could induce impulses of 1 bar loadintensity and 4 Hz frequency (4 cycles per second)on the bracket. The samples were submitted to 100000 mechanical cycles and 500 thermal cycles which varied from 5C to 55C. The SBS test was performed in a universal testing machine (1 mm/min). Afterdebonding, optic microscopic evaluation for adhesive penetration and Adhesive Remaining Index (ARI)analyses in the enamel surface were performed.The results showed higher SBS (14.70 ± 4.85 MPa)values for SEP. The SBS remained similar despitecycling (NC-11.44 MPa; TC-11.20 MPa; TMC-11.19MPa) for XT, meanwhile for SEP, ...

Systemic and microvascular effects of resuscitation with blood products after severe hemorrhage in rats.

BACKGROUND: Severe hemorrhage is associated with the disruption of the endothelial glycocalyx (EG), a key component of the endothelium. The effects of blood components on the EG are unknown. The present study furthers our investigations into the effects of resuscitation with blood products on the skeletal muscle microcirculation of hemorrhaged rats, focusing on packed red blood cells (PRBCs) or fresh whole blood (FWB). METHODS: Rats were bled 40% of total blood volume and resuscitated with 1:1 PRBC/lactated Ringer's solution (LR), 1:1 washed PRBC (wPRBC)/LR, FWB or LR only. Sham animals were subjected to all procedures except hemorrhage and resuscitation. EG thickness, blood flow, and microvascular permeability were studied using intravital microscopy. Hemodynamics and coagulation tests (rotational thromboelastometry) were performed. RESULTS: After severe hemorrhage, EG and permeability were restored to sham levels in the PRBC/LR and FWB groups, but not in the wPRBC/LR or LR groups. Clotting time was longer and clot elasticity and firmness were reduced in wPRBC/LR and LR, but not in FWB or PRBC/LR groups when compared with sham. CONCLUSION: Resuscitation with FWB or PRBC/LR was superior in reversing coagulopathy, restoring EG and permeability changes following hemorrhage, compared with wPRBC/LR or LR alone. As wPRBC/LR did not improve EG and permeability, these data suggest that the removal of residual plasma protein from wPRBC or resuscitation with a protein-free solution (LR) is not able to improve microcirculation and coagulation functions in this severe hemorrhage model.

Clostridium perfringens type A enteritis in blue and yellow macaw (Ara ararauna).

This study describes an outbreak of necrotic enteritis caused by Clostridium perfringens type A in captive macaws (Ara ararauna). Two psittacine birds presented a history of prostration and died 18 hr after manifestation of clinical signs. The necropsy findings and histopathologic lesions were indicative of necrotic enteritis. Microbiologic assays resulted in the growth of large gram-positive bacilli that were identified as C. perfringens. PCR was used to identify clostridium toxinotypes and confirmed the identification of isolated strains as C pefringens type A, positive to gene codifying beta 2 toxin. The infection source and predisposing factors could not be ascertained.

Effects of perfluorocarbon emulsions on microvascular blood flow and oxygen transport in a model of severe arterial gas embolism.

BACKGROUND: Arterial gas embolism (AGE) is a clinical problem that occurs directly in cardiopulmonary bypass machines in open-heart surgeries, or indirectly (through cardiac or pulmonary right to left shunts) in dive accidents, resulting in serious morbidity and even death. Perfluorocarbon (PFC) emulsions have been used for the treatment of AGE in an animal model. We hypothesized that PFC emulsions enhance microvascular blood flow, speed bubble resolution, and oxygenation in AGE compared with saline in a model of cremaster muscle from anesthetized rats. MATERIALS AND METHODS: AGE was induced by direct air injection into the femoral artery ipsilateral to the studied cremaster muscle. Microhemodynamics, microvascular, and tissue oxygenation were determined before and after treatment with two different commercial PFC emulsions (C10F20, Oxycyte; Oxygen Biotherapeutics, Inc and C10F18, PHER-O2; Sanguine Corporation, Inc) compared with saline in real time using brightfield and phosphorescence microscopy. RESULTS: Blood pressure and heart rate remained unchanged. Systemic PO2, oxygen (O2) content, and glucose were higher in PFC groups, whereas hematocrit dropped in all groups. Arteriolar blood flow went up 85% and 80% of baseline after C10F20 and C10F18 treatments, respectively, versus 11% after saline treatment. Arteriolar and tissue PO2, and O2 delivery were higher in PFC groups compared with the control group. There was an increase in arteriolar blood flow, reduction in diffusional resistance of O2 in the plasma, and improved tissue oxygenation. CONCLUSIONS: Administration of PFC emulsions in AGE is superior to saline primarily because of surfactant properties along with air bubble reabsorption.

Chronic exposure of lung alveolar epithelial type II cells to tobacco-specific carcinogen NNK results in malignant transformation: a new in vitro lung carcinogenesis model.

Lung cancer is the leading cause of cancer-related mortality in both men and women throughout the world. This disease is strongly associated with tobacco smoking. The aim of this manuscript was to establish an in vitro model that mimics the chronic exposures of alveolar epithelial type II cells to the tobacco-specific nitrosamine carcinogen, NNK. Immortalized non-neoplastic alveolar epithelial cells type II, (E10 cells), from BALB/c mice were exposed to low concentration of NNK (100 pM) during 5, 10, 15, and 20 cycles of 48 h. NNK-transformed cells showed an increase of proliferation rate and motility. Moreover, these cells underwent epithelial-to-mesenchymal transition (EMT). Increased migratory capacity and EMT were correlated to the time of exposure to NNK. NNK-transformed cells were tested for their growth and metastatic capacity in vivo. Subcutaneous injection of cells exposed to NNK for 20 cycles (E10-NNK20 clone) into BALB/c mice led to the formation of subcutaneous tumors that arose after 40 ± 17 d in all animals, which died 95 ± 18 d after cell inoculation, with lymph nodes and lung metastasis. The morphological characteristics of tumors were compatible with metastatic undifferentiated carcinoma. Cells exposed to NNK for 5-10 cycles did not display metastatic capacity, while those exposed for 15 cycles displayed low capacity. Our results show that prolonged exposures to NNK led the cells to increasingly acquire malignant properties. The cellular model presented in this study is suitable for studying the molecular events involved in the different stages of malignant transformation.