Ischemic myelomalacia and closed spinal dysraphism in multiple finishing swine.

Ischemic myelomalacia secondary to fibrocartilaginous emboli (FCE) is an idiopathic disease in humans and animals. On the other hand, congenital spinal cord malformations result from neural tube defects in fetal development (ie, spinal dysraphism), with structural anomalies referred to collectively as myelodysplasia. Spinal dysraphisms are frequently accompanied by skin and vertebral abnormalities because of the embryogenic relationship. In this observational case study, we report the pathologic findings of 13, 18- to 24-weeks-old pigs from a large conventional operation that presented with acute paraparesis. Ischemic myelomalacia secondary to FCE was observed in 5 of 13 examined pigs. Congenital spinal cord malformations located between the caudal thoracic and sacral spinal cord were identified in 7 pigs, with structural abnormalities that ranged from diplomyelia/split cord malformation to segmental spinal dysgenesis (myelodysplasia) to caudal agenesis. Concurrent myelomalacia and congenital spinal cord malformations in the same or different sites were noted in 2 pigs. No spinal lesion was observed in 3 pigs. Although gross vertebral abnormalities were not observed herein, intervertebral instability due to minor defects in the articular facets, as well as other unidentified factors, is suspected to contribute high incidence of FCE. It is likely that these congenital malformations were previously underdiagnosed or are possibly new conditions associated with continuous inbreeding and genetic improvement in the modern swine industry.

Successful conservative management in a dog with substantial urinary bladder ischemia.

A 7-year-old spayed female pug dog was brought to the veterinary college with a severely and diffusely ischemic urinary bladder secondary to obstructive uroliths in the lower urinary tract. Cystotomy was performed to remove the uroliths and the ischemic bladder was managed with conservative treatment. A recheck abdominal ultrasound 4.5 mo after surgery revealed an abdominal mass that was associated with the urinary bladder. An exploratory laparotomy and partial cystectomy were performed. Histopathology of the mass showed granulomatous inflammation centered on necrotic tissue. The dog recovered well, and long-term prognosis is good. To the best of our knowledge, this is the first veterinary case report describing conservative management of an ischemic urinary bladder. An uncommon complication following cystotomy and the relevant imaging findings is also described. The positive outcome for the dog demonstrated that conservative management may serve as an option for treatment of substantial ischemia of the urinary bladder.

Prise en charge conservatrice réussie chez un chien présentant une ischémie importante de la vessie. Une chienne carlin femelle stérilisée âgée de 7 ans a été amenée à l'école vétérinaire avec une vessie sévèrement et diffusément ischémique secondaire à des urolithes obstructifs dans les voies urinaires inférieures. Une cystotomie a été réalisée pour retirer les urolithes et la vessie ischémique a été prise en charge avec un traitement conservateur. Une échographie abdominale de contrôle 4,5 mois après la chirurgie a révélé une masse abdominale associée à la vessie. Une laparotomie exploratrice et une cystectomie partielle ont été réalisées. L'histopathologie de la masse a montré une inflammation granulomateuse centrée sur le tissu nécrotique. Le chien a bien récupéré et le pronostic à long terme est bon. À notre connaissance, il s'agit du premier rapport de cas vétérinaire décrivant la prise en charge conservatrice d'une vessie ischémique. Une complication peu fréquente après cystotomie et les résultats d'imagerie pertinents sont également décrits. Le résultat positif pour le chien a démontré qu'une gestion conservatrice peut servir d'option pour le traitement d'une ischémie importante de la vessie.(Traduit par Dr Serge Messier).

MicroRNAs are differentially expressed in the serum and renal tissues of cats with experimentally induced chronic kidney disease: a preliminary study.

OBJECTIVE: To identify differentially expressed microRNA in the serum and renal tissues of cats with experimentally induced chronic kidney disease (CKD). SAMPLE: Banked renal tissues and serum from 4 cats. PROCEDURES: Cats previously underwent 90-minute unilateral ischemia with delayed contralateral nephrectomy 3 months after ischemia. Tissues were collected from the contralateral kidney at the time of nephrectomy and from the ischemic kidney 6 months after nephrectomy (study end). Serum was collected prior to ischemia (baseline serum) and at study end (end point serum). Total RNA was isolated from tissues and serum, and microRNA sequencing was performed with differential expression analysis between the contralateral and ischemic kidney and baseline and end point serum. RESULTS: 20 microRNAs were differentially expressed between ischemic and contralateral kidneys, and 52 microRNAs were differentially expressed between end point and baseline serum. Five microRNAs were mutually differentially expressed between ischemic and contralateral kidneys and baseline and end point serum, with 4 (mir-21, mir-146, mir-199, and mir-235) having increased expression in both the ischemic kidney and end point serum and 1 (mir-382) having increased expression in the ischemic kidney and decreased expression in end point serum. Predicted target search for these microRNA revealed multiple genes previously shown to be involved in the pathogenesis of feline CKD, including hypoxia-inducible factor-1α, transforming growth factor-ß, hepatocyte growth factor, fibronectin, and vascular endothelial growth factor A. CLINICAL RELEVANCE: MicroRNAs were differentially expressed after CKD induction in this preliminary study. Regulation of renal fibrosis in feline CKD may occur through microRNA regulation of mRNAs of pro- and anti-fibrotic genes.

Pin1 aggravates renal injury induced by ischemia and reperfusion in rats via Nrf2/HO-1 mediated endoplasmic reticulum stress

Purpose: To investigate the role of peptidyl-prolyl cis/trans isomerase 1 (Pin1) on renal ischemia-reperfusion (I/R) injury and underlying mechanism. Methods: By establishing the in vitro and in vivo models of renal I/R, the role of Pin1 was explored by using molecular assays. Results: In renal I/R, endogenous Pin1 level was up-regulated in I/R-impaired kidney. Suppression of Pin1 with juglone afforded protection against I/R-mediated kidney dysfunction, and reduced I/R-induced endoplasmic reticulum (ER) stress in vivo. Consistent with the in vivo results, repression of Pin1 with juglone or gene knockdown with si-Pin1 conferred cytoprotection and restricted hypoxia/reoxygenation (H/R)-driven ER stress in HK-2 cells. Simultaneously, further study uncovered that Nrf-2/HO-1 signals was the association between Pin1 and ER stress in response to renal I/R. In addition, Nrf-2/HO-1 signal pathway was inactivated after kidney exposed to I/R, as indicated by the down-regulation of Nrf-2/HO-1 levels. Furthermore, inhibition of Pin1 remarkably rescued the inactivation ofNrf-2/HO-1. Conclusions: Pin1 modulated I/R-mediated kidney injury in ER stress manner dependent on Nrf2-HO-1 pathway in I/R injury.

Ability of different assay platforms to measure renal biomarker concentrations during ischaemia-reperfusion acute kidney injury in dogs.

Several protein biomarkers have been shown to be useful for the early diagnosis of acute kidney injury (AKI) in animals and people. Multiplex assays for measurement of a panel of renal biomarkers in canine samples have recently become available. This study compared the use of two such assays, versus previously validated ELISAs, to measure five biomarkers in canine samples during ischaemia-reperfusion (IR) AKI. Blood and urine was collected from six male anaesthetised greyhounds that underwent 1-h of renal ischaemia (severe hypotension induced by acute haemorrhage) and 2-h of reperfusion (intravenous fluid resuscitation). Histology confirmed presence of acute tubular injury at 2 h of reperfusion. Concentrations of clusterin, cystatin C, kidney-injury molecule 1 (KIM-1), monocyte chemoattractant protein 1, and neutrophil gelatinase-associated lipocalin (NGAL) at baseline and following IR, measured by two different multiplex assays and previously-validated single analyte immunoassays, were compared. Only NGAL was significantly elevated following IR with all assays investigated. Whether concentrations of the other four biomarkers were significantly increased following IR depended on the assay used. Concentrations of cystatin C and KIM-1 measured with the multiplex assays were of a vast magnitude lower than those measured with the corresponding single analyte ELISAs. We conclude that further validation is required before these assays can reliably be used to measure AKI biomarkers in canine samples.

The clinical efficacy of pentoxifylline and l-glutamine on ischemia and reperfusion injury in cattle with displaced abomasum: a longitudinal study.

This study aimed to assess the clinical efficacy of pentoxifylline (PTX) and L-glutamine (L-Gln) treatment on ischemia and reperfusion (I/R) injury in the abomasal tissue, acute phase response (APR), oxidative stress (OS), cytokine response, hemostatic, and coagulation disorders in the 96-h period before and after surgery in displaced abomasum (DA) cases. The study sample consisted of 48 dairy cows with DA that were categorized into four groups as group S (Sham group) (9 Left displaced abomasum (LDA)+3 Right displaced abomasum (RDA), group P (PTX) (10 LDA+2 RDA), group G (L-Gln) (10 LDA+2 RDA), and group P+G (PTX+L-Gln) (10 LDA+2 RDA). Acute-phase protein (Haptoglobin), oxidative stress indicators (malondialdehyde, nitric oxide, and glutathione), cytokines (tumor necrosis factor (TNF)-α and interleukin-1ß (IL-1ß), coagulation factors (D-Dimer, Antithrombin (ATIII), Thrombin-antithrombin complex, Plasminogen activator inhibitor-1), and enzyme activities (lactate dehydrogenase, gamma- -glutamyl transferase, sorbitol dehydrogenase, glutamate dehydrogenase, adenosine deaminase, myeloperoxidase, and creatine phosphokinase) in blood serum samples and coagulometric analyses of blood plasma were performed in samples taken before the operation and at 30 and 60 min and 2, 5, 10, 24, 48, 72, and 96 h after the operation. In DA cases, while post-operative treatment procedures with PTX and L-Gln were effective in decreasing APR and OS, these were ineffective in prohibiting the inflammatory response coordinated by cytokines. For the treatment and prevention of I/R injury in the DA cases, PTX and L-Gln procedures hold promise with their effects on APR, OS, and hemostatic dysfunction. Additional treatment procedures are required for the suppression of inflammatory response, and the effectiveness of preconditioning treatment may be evaluated.

Brief topical and intraluminal use of Carolina rinse solution does not attenuate experimental ischemia and reperfusion injury in rabbit jejunum / [O uso tópico breve e intraluminal da solução de Carolina rinse não atenua a injúria de isquemia e reperfusão experimental no jejuno de coelhos ]

Fifteen New Zealand adult rabbits were randomly allocated into three groups: Sham-operated (group A), Ischemia and Reperfusion (group B) and Carolina Rinse Solution (CRS) (group C). Groups B and C were subjected to one hour of ischemia and two hours of reperfusion. In group C, ten minutes before reperfusion, the bowel lumen was filled with CRS, and the segment immersed in CRS. Necrosis and loss of integrity of the villi were visible in groups B and C. Edema of the submucosa and circular muscle was observed in all groups. Hemorrhage was observed in different layers for groups B and C, but group C showed more severe hemorrhage in different layers during reperfusion. All groups showed polymorphonuclear leukocyte infiltration on the base of the mucosa, submucosa, and longitudinal muscle, in addition to polymorphonuclear leukocytes margination in the mucosal and submucosal vessels. Necrosis of enterocytes, muscles, crypts of Lieberkühn and myenteric plexus was observed in groups B and C during reperfusion. Topical and intraluminal Carolina Rinse Solution did not attenuate the effects of ischemia and reperfusion in the small intestine of rabbits.(AU)

Quinze coelhos da raça Nova Zelândia foram alocados em três grupos: instrumentado (grupo A), isquemia e reperfusão (grupo B) e solução de Carolina rinse (CRS) (grupo C). Os grupos B e C foram submetidos a uma hora de isquemia e a duas horas de reperfusão. No grupo C, 10 minutos antes da reperfusão, o segmento isolado foi imerso e teve seu lúmen preenchido com CRS. Os grupos B e C apresentaram necrose e perda progressiva da integridade das vilosidades. Foi observado edema na submucosa e na camada muscular circular em todos os grupos. Nos grupos B e C, foi observada hemorragia em diferentes camadas, mas, no grupo C, a hemorragia foi mais intensa durante a reperfusão. Todos os grupos apresentaram infiltrado de PMN na base da mucosa, na submucosa e na camada muscular longitudinal e marginação de PMN nos vasos da mucosa e da submucosa. Durante a reperfusão, foi observada necrose dos enterócitos, das camadas musculares, das criptas de Lieberkühn e do plexo mioentérico nos grupos B e C. O uso tópico e intraluminal de CRS não atenuou os efeitos da isquemia e da reperfusão no intestino delgado de coelhos.(AU)

Two-dimensional and contrast-enhanced ultrasound of intestinal ischaemia in cats: four cases.

OBJECTIVES: The aim of this study was to describe the results of two-dimensional (2D) and contrast-enhancement ultrasound (CEUS) in four cats with intestinal ischaemia. METHODS: Data were collected from hospital records of all cats that had intestinal ischaemia between January 2012 and August 2018. The inclusion criteria were complete abdominal ultrasound examination, colour flow Doppler and CEUS of lesions, confirmation of intestinal ischaemia detected by visual assessment of avascular intestinal segment at surgery, and/or necropsy and histopathology. All images and video clips were reviewed by the same experienced operator. RESULTS: Four cats with different intestinal ischaemic lesions were included in the study: duodenal perforating ulcer, jejunal necrotising enteritis, necrosis secondary to jejunojejunal intussusception and iatrogenic damage of jejunal arteries. On the 2D ultrasound, all intestinal lesions were characterised by non-specific findings: focal hypoechoic wall thickening with loss of normal layering associated with hyperechoic mesentery surrounding the intestinal tract. CEUS showed a reduced or absent enhancement of the intestinal lesions in comparison to the surrounding perfused wall. CONCLUSIONS AND RELEVANCE: Intestinal ischaemia is a potentially fatal disorder. Grey-scale, colour and power Doppler ultrasonography are not sensitive for evaluating this condition. Our preliminary findings illustrate the usefulness of CEUS for the detection of intestinal wall impaired perfusion in cats.

Temporal changes in urinary excretion of liver-type fatty acid binding protein (L-FABP) in acute kidney injury model of domestic cats: a preliminary study.

Liver-type fatty acid-binding protein (L-FABP) is a biomarker for the early detection of renal diseases in humans. It is secreted along with cytotoxic oxidation products from proximal tubular epithelial cells under conditions of ischemia and/or oxidative stress. This study examined urinary L-FABP excretion under renal ischemia in feline acute kidney injury (AKI) model. L-FABP excretion increased immediately after renal ischemia/reperfusion, despite the absence of obvious structural damage to the kidneys, in the two AKI model cats studied. L-FABP was detected in the renal tubular lumen immediately after renal ischemia/reperfusion in the two cats, but not in a sham surgery cat. These results suggested that high L-FABP excretion is a pathophysiological response associated with antioxidant defense in proximal tubules with renal ischemia and/or oxidative stress in a feline model.

Canine ischaemic dermatopathy: a retrospective study of 177 cases (2005-2016).

BACKGROUND: Ischaemic dermatopathy encompasses a poorly understood subset of canine diseases that share similar clinical and histological features. Very little information is currently available regarding population characteristics, progression and outcome. HYPOTHESIS/OBJECTIVES: This study aimed to describe the clinical features and therapeutic outcomes of ischaemia dermatopathy, excluding familial dermatomyositis, using cases diagnosed by histopathological analysis. ANIMALS: One hundred and seventy-seven cases submitted for histopathological analysis between 2005 and 2016 met inclusion criteria, of which 93 had complete medical records available. METHODS AND MATERIALS: Both records and pointed surveys were used to retrieve information. Scoring systems were created to subjectively evaluate clinical outcomes and likelihood of a vaccine association. RESULTS: Of 177 cases, toy and miniature poodles, Chihuahuas, Maltese, Yorkshire terriers and Jack Russell terriers were significantly over-represented (P < 0.001). Of the 93 cases for which historical data were obtained, median age at skin biopsy was five years (0.42-13 years) and median body weight was 7.3 kg (range 1.32-50.3 kg). The condition in 45 dogs (48.3%) was found likely to be associated with vaccination. Younger ages (P = 0.011) and higher body weights (P = 0.003) were positively correlated with greater likelihood of vaccination. Body weight <10 kg (P = 0.0045) and older ages (P = 0.0048) were significantly associated with worse outcomes. CONCLUSIONS AND CLINICAL IMPORTANCE: This study provides support for breed predispositions and identifies potential prognostic factors. Importantly, over half of the cases were considered unlikely to be vaccine-associated, demonstrating the need to investigate other underlying causes of this condition.

Allogenic mesenchymal stem cell intravenous infusion in reparation of mild intestinal ischemia/reperfusion injury in New Zealand rabbits / Infusão intravenosa de células tronco mesenquimais alógenas na reparação da lesão branda de isquemia/reperfusão intestinal em coelhos Nova Zelândia

The present study aimed to evaluate the efficacy of mesenchymal stem cell (MSC) infusion, derived from adipose tissue, on reduction of local and remote tissue damage caused by the event of experimental intestinal I/R in New Zealand breed rabbits. For obtaining, characterization, and cultivation of MSC derived from adipose tissue (MSC-Adp), 3 juvenile animals (four months old) were used. The cells were considered to be viable for therapy after the fourth passage (in vitro phase). For the in vivo stage, 24 young adult animals (six months old) were used, weighing approximately 3.5 kg, in which were randomly divided into two groups, called: IR treated with MSC (I2H/R5H MSC 3D; I2H/R5H MSC 7D); IR treated with PBS (I2H/R5H PBS 3D; I2H/R5H PBS 7D). The animals were anesthetized and submitted to pre-retro-umbilical midline celiotomy. The extramural peri-intestinal marginal artery was located and clamped (predetermined and standardized region) with the aid of a vascular clip, promoting a 2 hour blood flow interruption. After this period, blood flow was reestablished, inhalatory anesthesia was suspended, and the animals awaken. After 5 hours of reperfusion, the treatments were performed by intravenous infusion according to the experimental groups. The animals were evaluated 72 hours and seven days after the treatment as for the macroscopic appearance (color and peristaltism) of the jejunal segment, and by histological evaluation of the ischemic segment for the presence or absence of destruction of the intestinal mucosa, edema, bleeding, dilation of lymph vessels, and presence of polymorphonuclear inflammatory cells, both in the mucosa and submucosa. The observed results revealed that the groups treated with MSC-Adp obtained smaller mucosal and submucosal lesions when compared to the groups treated with PBS. Also, MSC-Adp treated groups obtained controlled inflammatory response and higher mitotic rate, outcomes related to the therapeutic potential of MSC. Infusion of stem cells attenuated the lesions caused by intestinal I/R in both MSC groups when compared to the group treated with PBS.(AU)

O presente estudo teve como principal objetivo avaliar a eficácia da infusão células tronco mesenquimais (CTM) derivada de tecido adiposo sobre diminuição das lesões teciduais locais e remotas, causadas pelo evento de I/R intestinal experimental, em coelhos da raça Nova Zelândia. Para obtenção, cultivo e caracterização das CTM provenientes de tecido adiposo (ADCTM) foram utilizados 3 animais jovens. As células foram consideradas viáveis para terapia a partir da quarta passagem (fase in vitro). Para etapa in vivo foram utilizados 24 animais, adulto-jovens, pesando aproximadamente 3,5kg, divididos aleatoriamente em dois grupos experimentais, denominados IR Tratado com CTM (I2H/R5H CTM 3D; I2H/R5H CTM 7D); IR Tratado PBS (I2H/R5H PBS 3D; I2H/R5H PBS 7D). Os animais foram anestesiados e submetidos à celiotomia mediana pré-retroumbilical. A artéria marginal peri-intestinal extramural foi localizada e clampeada (região predeterminada e padronizada) com auxílio de um clipe vascular, promovendo uma interrupção do fluxo sanguíneo durante 2 horas. Após esse período, o fluxo sanguíneo foi restabelecido, a anestesia inalatória suspendida e os animais despertados. Após 5 horas de reperfusão realizou-se os tratamentos por infusão endovenosa, conforme grupos experimentais. Os animais foram avaliados 72 horas e sete dias após o tratamento quanto ao aspecto macroscópico (coloração e peristaltismo) do segmento jejunal e por meio de avaliação histológica do segmento isquemiado quanto à presença ou ausência de destruição de mucosa intestinal, edema, hemorragia, dilatação de vasos linfáticos e presença de células inflamatórias polimorfornucleares, tanto em mucosa quanto submucosa. Os resultados observados revelaram que os grupos tratados com ADCTM obtiveram menores lesões em mucosa e submucosa quando comprados aos grupos tratados com PBS. Ainda os grupos tratados com ADCTM obtiveram resposta inflamatória controlada e maior taxa mitótica, resultados relacionados ao potencial terapêutico das CTM.(AU)

Pilot study: use of contrast-enhanced ultrasonography in feline renal transplant recipients.

Objectives The study aims were to evaluate the feasibility of contrast-enhanced ultrasonography in feline renal transplant recipients in the post-transplantation period and to report findings in a case with presumptive delayed allograft ischemia. Methods Cats were imaged postoperatively using contrast harmonic ultrasonography after a bolus injection of a microbubble contrast medium. Time/mean pixel intensity curves were generated for cortical and medullary regions of interest in the renal allograft in each cat. Arrival time, time to peak, wash-in slope, wash-out slope, mean transit time and renal blood flow were calculated for each area. Results Within the renal cortices of cats without ureteral obstruction 1 day post-transplantation, arrival time was 2.0-6.3 s, time to peak was 3.6-30.1 s, wash-in rate was 2.45-41.14 mean pixel intensity (MPI)/s, wash-out rate was -2.01 to -0.47 MPI/s and blood flow was 6.1-106.5 MPI/s. Ratio mean transit time was 0.29-1.29. Typical cortical and medullary perfusion patterns were observed in these cats. In one cat with delayed graft ischemia followed by presumptive acute transplant rejection, dynamic and heterogeneous cortical and medullary perfusion was demonstrated. Decreases in cortical blood flow were paralleled by elevated serum creatinine. Conclusions and relevance Contrast-enhanced ultrasonography can be used in feline renal transplant recipients and provides both qualitative and quantitative data regarding renal allograft perfusion.

Histological evaluation and E-cadherin and ß-catenin expression in kidney of dogs submitted to renal ischemia and reperfusion after chlorpromazine administration / Avaliação histológica e imunomarcação de E-caderina e ß-catenina em rins de cães submetidos a isquemia e referfusão renal após administração de clorpromazina

Renal ischemia can be associated with some urological procedures, such as renovascular surgery or kidney transplantation, that are often followed by acute renal failure. The aim of this study was to verify the E-cadherin and ß-catenin localization in canine kidney in different times of renal ischemia and reperfusion after chlorpromazine application. Twelve dogs were randomly distributed equally into two groups. GroupA with ischemia and reperfusion without chlorpromazine and groupB with ischemia and reperfusion treated by chlorpromazine. GroupB received intravenous chlorpromazine, 15 min before the artery obstruction, which lasted 1 hour. After this period, the clamps in the renal arteries were released and the organ remained in reperfusion for 2 hours. In each group, anti-E-cadherin and anti-ß-catenin antibodies were made in six tissue samples from renal parenchyma. E-cadherin and ß-catenin are differentially expressed in segments from cortex and medulla in dog's kidneys and the use of chlorpromazine did not alter the expression of both proteins. Occlusion of the left renal artery in dogs causes morphological alterations mainly in proximal convoluted tubules, beginning 30min after the start of ischemia and being aggravated after two hours of reperfusion. These results reveal that chlorpromazine did not change kidneys' histological aspect nor E-cadherin and ß-catenin expression.(AU)

A lesão renal isquêmica pode estar associada a procedimentos urológicos, tais como cirurgia renovascular, cirurgia renal extracorpórea ou transplante renal. Essa injúria, muitas vezes, é seguida de insuficiência renal aguda. O objetivo deste trabalho foi observar a localização da E-caderina e da ß-catenina em rim de cães, além de relacionar a expressão dessas proteínas das junções de aderência em diferentes tempos de isquemia e reperfusão com ou sem a aplicação de clorpromazina. Para tanto, foram utilizados 12 cães, distribuídos aleatoriamente em dois grupos de seis indivíduos: grupo A, com isquemia e reperfusão sem tratamento por clorpromazina, e o grupo B, com isquemia e reperfusão tratado por clorpromazina. No procedimento cirúrgico, foi realizada uma incisão paracostal esquerda para identificação e isolamento do rim esquerdo e da artéria renal esquerda. Após o isolamento da artéria, os animais de todos os grupos tiveram o vaso ocluído. Os animais do grupo B receberam clorpromazina via endovenosa, na dose de 5mg/kg, 15min antes da clampagem do vaso, que durou uma hora. Após este período, as artérias renais foram desobstruídas e os órgãos permaneceram em reperfusão por duas horas. Em cada grupo, foram extraídas seis amostras de parênquima renal, com utilização de agulha tru-cut, para marcação com anticorpos anti-E-caderina e anti-ß-catenina por meio de imunoistoquímica. E-caderina e ß-catenina são diferencialmente expressas em segmentos do córtex e da medula em rim de cães e o uso da clorpromazina não alterou a expressão das duas proteínas.(AU)

Optimizing mesoderm progenitor selection and three-dimensional microniche culture allows highly efficient endothelial differentiation and ischemic tissue repair from human pluripotent stem cells.

BACKGROUND: Generation of large quantities of endothelial cells is highly desirable for vascular research, for the treatment of ischemia diseases, and for tissue regeneration. To achieve this goal, we developed a simple, chemically defined culture system to efficiently and rapidly differentiate endothelial cells from human pluripotent stem cells by going through an MESP1 mesoderm progenitor stage. METHODS: Mesp1 is a key transcription factor that regulates the development of early cardiovascular tissue. Using an MESP1-mTomato knock-in reporter human embryonic stem cell line, we compared the gene expression profiles of MESP1+ and MESP1- cells and identified new signaling pathways that may promote endothelial differentiation. We also used a 3D scaffold to mimic the in vivo microenvironment to further improve the efficiency of endothelial cell generation. Finally, we performed cell transplantation into a critical limb ischemia mouse model to test the repairing potential of endothelial-primed MESP1+ cells. RESULTS: MESP1+ mesoderm progenitors, but not MESP1- cells, have strong endothelial differentiation potential. Global gene expression analysis revealed that transcription factors essential for early endothelial differentiation were enriched in MESP1+ cells. Interestingly, MESP1 cells highly expressed Sphingosine-1-phosphate (S1P) receptor and the addition of S1P significantly increased the endothelial differentiation efficiency. Upon seeding in a novel 3D microniche and priming with VEGF and bFGF, MESP1+ cells markedly upregulated genes related to vessel development and regeneration. 3D microniches also enabled long-term endothelial differentiation and proliferation from MESP1+ cells with minimal medium supplements. Finally, we showed that transplanting a small number of endothelial-primed MESP1+ cells in 3D microniches was sufficient to mediate rapid repair of a mouse model of critical limb ischemia. CONCLUSIONS: Our study demonstrates that combining MESP1+ mesoderm progenitor cells with tissue-engineered 3D microniche and a chemically defined endothelial induction medium is a promising route to maximizing the production of endothelial cells in vitro and augment their regenerative power in vivo.

Laminar lesions in horses with systemic oxidative stress, committed by experimentally induced or naturally occurring gastrointestinal disorders / Lesões laminares em cavalos com estresse oxidativo sistêmico, acometidos por doenças gastrintestinais experimentalmente induzidas ou de ocorrência natural

Laminitis in horses can be associated with lesions in multiple organs secondary to sepsis. Twenty-one horses suffering from gastrointestinal disorders were used in the experiment; 7 horses with experimentally induced endotoxemia and intestinal ischaemia, and 14 horses suffering from naturally occurring colic syndrome. Tissue samples of lungs, liver, heart, brain, cerebellum and hoof laminar tissue were collected for histopathological and oxidative stress evaluation using nitrotyrosine and superoxide dismutase (SOD2) immunostaining. The horses were divided into two groups: the non-oxidative lesions group (NOLG), with 7 horses showing weak immunostaining in lungs, liver and kidney, and the oxidative lesions group (OLG), with 14 horses showing immunostaining indicating systemic oxidative stress in multiple organs. The horses from OLG showed increase of laminar lesions and SOD2 immunostaining in multiple organs when compared to the horses from the NOLG. No differences were found ln regard to laminar immunostaining by nitrotyrosine and SOD2 between experimental groups. It was concluded that systemic oxidative stress can be associated with the development of laminar lesions, and that the laminar tissue does not respond to oxidative stress with increase of SOD as occurs in other organs.(AU)

A laminite em equinos pode estar associada à lesão em múltiplos órgãos secundária a sepse. Foram utilizados 21 cavalos com afecções gastrintestinais, sendo sete com endotoxemia e isquemia intestinal induzidos experimentalmente, e 14 cavalos com síndrome cólica de origem natural. Amostras teciduais de pulmão, rim, fígado, coração, cérebro e cerebelo e de tecido laminar do casco foram coletadas para avaliação de lesão histopatológica e estresse oxidativo, pela imunomarcação de nitrotirosina e superóxido dismutase (SOD2). Os animais foram divididos em dois grupos: grupo sem lesão oxidativa (NOLG), com sete cavalos com fraca imunomarcação em pulmão, fígado e rim, e grupo lesão oxidativa (OLG), contendo 14 cavalos com imunomarcação indicando estresse oxidativo em múltiplos órgãos. Os cavalos do grupo OLG apresentaram aumento de lesões laminares e imunomarcação para SOD2 em múltiplos órgãos, quando comparados ao NOLG. Não houve diferença sobre a imunomarcação laminar para nitrotirosina e SOD2 entre os grupos experimentais. Conclui-se que o estresse oxidativo sistêmico está associado ao desenvolvimento de lesões laminares, e que o tecido laminar não responde ao estresse oxidativo com aumento de SOD como ocorre nos outros órgãos.(AU)

Heart rate variability in horses with acute gastrointestinal disease requiring exploratory laparotomy.

OBJECTIVE: To describe heart rate variability (HRV) in horses with acute gastrointestinal disease that undergo exploratory laparotomy. We hypothesized that horses with ischemic gastrointestinal disease will have reduced HRV compared to horses with nonischemic lesions. We further hypothesized that a reduction in HRV will be associated with nonsurvival. DESIGN: Prospective, clinical, observational study. SETTING: University veterinary teaching hospital. ANIMALS: Horses presented for acute colic (n = 57) or elective surgical procedures (n = 10) were enrolled. INTERVENTIONS: Admission heart rate (HR) was recorded and within 2 hours of recovery from general anesthesia continuous telemetry was placed, monitored and recorded for 48-52 hours postoperatively. Stored electrocardiograms were manually inspected and R-to-R intervals were extracted and uploaded into HRV software for analysis. Time domain and frequency spectral analysis were investigated at Times 1 (2-10 h), 2 (16-24 h), 3 (30-38 h), and 4 (44-52 h) postoperatively. A two-way ANOVA for repeated measures was used for group comparisons. Logistic regression analysis was used to detect potential associations between admission HR, time and frequency domain variables, and nonsurvival. MEASUREMENTS AND MAIN RESULTS: Horses diagnosed with an ischemic gastrointestinal lesion (n = 22) at the time of surgery had significantly higher postoperative heart rates and reduced time domain-derived measures of HRV than horses with nonischemic gastrointestinal lesions (n = 35) or control horses (n = 10). Horses that survived to discharge had significantly lower postoperative HRs, higher time domain, and lower low frequency spectral measures of HRV compared to nonsurvivors. The multivariable logistic regression model had a receiver operating characteristic area under the curve (AUC) of 0.95 and was significantly better at predicting nonsurvival than admission HR (P = 0.0124). CONCLUSIONS: Reduced HRV was strongly associated with ischemic gastrointestinal disease and nonsurvival. HRV analysis is a noninvasive technique that may provide diagnostic and prognostic information pertinent to the management of postoperative horses with severe gastrointestinal disease.

Idiopathic ischemic necrosis of an accessory carpal bone in a dog.

CASE DESCRIPTION: A 6-year-old neutered female mixed-breed dog was evaluated because of a 6-week history of left forelimb lameness that varied in severity. CLINICAL FINDINGS: Radiography revealed expansile and lytic changes of the left accessory carpal bone (ACB). Results of histologic evaluation of ACB core biopsy specimens indicated areas of bone necrosis. The entire left ACB was excised and submitted for histologic evaluation; results confirmed a diagnosis of idiopathic ischemic necrosis. TREATMENT AND OUTCOME: Left pancarpal arthrodesis was performed to treat carpal hyperextension and persistent lameness. The dog had an excellent functional outcome with no other problems related to the carpus until its death 4 years later, further decreasing suspicion that the problem was attributable to an undetected neoplasm or bacterial or fungal osteomyelitis. CLINICAL RELEVANCE: The radiographic and histologic findings for the dog of this report were similar to previously reported findings for dogs with ischemic femoral head necrosis and humans with ischemic carpal (pisiform or lunate bone) necrosis. The etiology of the ischemic ACB necrosis in this dog was not determined. To the authors' knowledge, this is the first report of a dog with idiopathic ischemic ACB necrosis. Idiopathic ischemic necrosis should be included as a differential diagnosis for dogs with lameness and destructive and expansile ACB radiographic lesions. An excellent functional outcome may be attained by means of ACB excision and pancarpal arthrodesis.

Investigation of muscle degeneration process in young rats with ischemia injury using MR diffusion tensor imaging.

Skeletal muscle is vulnerable to ischemia injury after direct trauma or indirect causes. Magnetic resonance diffusion tensor imaging (DTI) has been demonstrated to be a powerful tool to investigate muscle structures. However, most previous DTI studies that investigated ischemia muscle were performed on mature species, results of which may not apply on other age populations, such as young groups. In this study, age-related variation of muscle regeneration course after ischemia injury was investigated for the first time. Ten young and ten mature SD rats were induced ischemia in hindlimbs, and the evolutions of the skeletal muscle regeneration were longitudinally studied using DTI before and till 21 days after surgery. Results showed that the young group was more sensitive to ischemia and recovered more rapidly than the mature one. This study confirms the age-related variation of muscle regeneration process, and may provide supplemental information for better understanding of muscle repair evolution.

Ischaemic distal limb necrosis and Klebsiella pneumoniae infection associated with arterial catheterisation in a cat.

This case report describes dorsal pedal arterial thrombosis and infection with Klebsiella pneumoniae subsequent to arterial catheter placement in a cat. The complication led to avascular necrosis of the metatarsal and pedal soft tissue. The catheter was placed for blood pressure monitoring during surgery for correction of a peritoneopericardial diaphragmatic hernia. The exact mechanism of thrombosis was unclear. Amputation of the limb was required and the histopathological findings are presented. This is the first report of such a complication.