In Vitro Transdifferentiation Potential of Equine Mesenchymal Stem Cells into Schwann-Like Cells.

Schwann cells (SCs) are essential for the regenerative processes of peripheral nerve injuries. However, their use in cell therapy is limited. In this context, several studies have demonstrated the ability of mesenchymal stem cells (MSCs) to transdifferentiate into Schwann-like cells (SLCs) using chemical protocols or co-culture with SCs. Here, we describe for the first time the in vitro transdifferentiation potential of MSCs derived from equine adipose tissue (AT) and equine bone marrow (BM) into SLCs using a practical method. In this study, the facial nerve of a horse was collected, cut into fragments, and incubated in cell culture medium for 48 h. This medium was used to transdifferentiate the MSCs into SLCs. Equine AT-MSCs and BM-MSCs were incubated with the induction medium for 5 days. After this period, the morphology, cell viability, metabolic activity, gene expression of glial markers glial fibrillary acidic protein (GFAP), myelin basic protein (MBP), p75 and S100ß, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and glial cell-derived neurotrophic factor (GDNF), and the protein expression of S100 and GFAP were evaluated in undifferentiated and differentiated cells. The MSCs from the two sources incubated with the induction medium exhibited similar morphology to the SCs and maintained cell viability and metabolic activity. There was a significant increase in the gene expression of BDNF, GDNF, GFAP, MBP, p75, and S100ß in equine AT-MSCs and GDNF, GFAP, MBP, p75, and S100ß in equine BM-MSCs post-differentiation. Immunofluorescence analysis revealed GFAP expression in undifferentiated and differentiated cells, with a significant increase in the integrated pixel density in differentiated cells and S100 was only expressed in differentiated cells from both sources. These findings indicate that equine AT-MSCs and BM-MSCs have great transdifferentiation potential into SLCs using this method, and they represent a promising strategy for cell-based therapy for peripheral nerve regeneration in horses.

Applicability of tactile memory examination as an option to visual- and verbal-based batteries / A aplicabilidade da avalição da memória tátil como uma opção para as baterias visuais a verbais

ABSTRACT The validity and applicability of tactile battery tests for the diagnosis and medical follow-up of patients with Alzheimer's disease and other dementia syndromes do not have their usefulness well understood in clinical practice. While haptic abilities in older individuals receive less attention, in earlier stages of human life they are well focused on. There are even fewer studies on tactile memory, including episodic memory of demented individuals with or without sensorial limitations. The applicability of a new haptic memory battery was evaluated in patients with Alzheimer's disease with mild or moderate commitment. Objective: The aim of this study is to apply a battery based on tactile perception, recognition, and recollection of everyday objects in patients with Alzheimer's disease, testing tactile delayed recall memory discrimination and late recognition to compare validated visual and verbal tests. Methods: Tactile-, visual-, and verbal-based memory performance was registered in 21 patients diagnosed with Alzheimer's disease. Results: Except for tactile identification, it showed that there was a close relationship between the three sensory modalities of memory, with an apparent better performance of tactile incidental memory and recognition compared with the test with pictures. Conclusions: The haptic evaluation of memory demonstrated applicability in the evaluation of memory dysfunction in patients with Alzheimer's disease. Further studies are needed to establish the sensibility and specificity of the proposed test that had a small sample size and many limitations.

RESUMO A aplicação de baterias baseadas em informações táteis para diagnóstico e acompanhamento de pacientes com doença de Alzheimer e outras demências não é muito difundida na prática clínica. Ao passo que a capacidade de reconhecimento tátil de objetos em crianças recebe bastante atenção dos pesquisadores, o mesmo não ocorre com indivíduos mais idosos. Existem ainda menos estudos abordando avaliação de memória episódica em pacientes idosos, com ou sem limitações sensoriais, como cegueira e surdez. O presente estudo propõe avaliar aplicabilidade de teste para avaliação de memória tátil em paciente com doença de Alzheimer. Objetivo: Aplicar em pacientes com doença de Alzheimer uma bateria baseada na percepção e reconhecimento táctil de objetos de uso cotidiano, avaliando a memória e reconhecimento tardio, comparado com testes visuais e verbais validados. Métodos: Foi avaliado o desempenho de baterias baseadas em informação táteis, visuais e verbais em 21 pacientes diagnosticados com a doença de Alzheimer. Resultados: Com a exceção da percepção tátil, foi demonstrada uma boa correlação entre as três modalidades de apresentação das informações a serem resgatadas tardiamente. Houve um desempenho aparentemente melhor do reconhecimento tátil quando comparado com apresentação de figuras impressas. Conclusões: A determinação da memória tátil em pacientes com doença de Alzheimer demonstrou ser aplicável como instrumento de avaliação dos distúrbios de memória presentes em portadores de doença de Alzheimer com comprometimento leve a moderado. As limitações do estudo, com reduzido tamanho da amostra, aponta para a necessidade de novos estudos com a devida validação, incluindo determinação de sensibilidade e especificidade, assim como pareamento, de acordo com sexo e idade com grupo controle.

Intrathecal Transplantation of Autologous and Allogeneic Bone Marrow-Derived Mesenchymal Stem Cells in Dogs.

The route used in the transplantation of mesenchymal stem cells (MSCs) can directly affect the treatment success. The transplantation of MSCs via the intrathecal (IT) route can be an important therapeutic strategy for neurological disorders. The objective of this study was to evaluate the safety and feasibility of the IT transplantation of autologous (Auto-MSCs) and allogeneic (Allo-MSCs) bone marrow mesenchymal stem cells (BM-MSCs) in healthy dogs. Based on neurodisability score, cerebrospinal fluid (CSF) and magnetic resonance imaging (MRI), no significant differences from the control group were observed on day 1 or day 5 after IT Auto- or Allo-MSCs transplantation (P > 0.05). In addition, analysis of matrix metalloproteinase (MMP)-2 and MMP-9 expression in the CSF revealed no significant differences (P > 0.05) at 5 days after IT transplantation in the Auto- or Allo-MSCs group when compared to the control. Intrathecal transplantation of BM-MSCs in dogs provides a safe, easy and minimally invasive route for the use of cell-based therapeutics in central nervous system diseases.

3D-printed nerve guidance conduits multi-functionalized with canine multipotent mesenchymal stromal cells promote neuroregeneration after sciatic nerve injury in rats.

BACKGROUND: Nerve injuries are debilitating, leading to long-term motor deficits. Remyelination and axonal growth are supported and enhanced by growth factor and cytokines. Combination of nerve guidance conduits (NGCs) with adipose-tissue-derived multipotent mesenchymal stromal cells (AdMSCs) has been performing promising strategy for nerve regeneration. METHODS: 3D-printed polycaprolactone (PCL)-NGCs were fabricated. Wistar rats subjected to critical sciatic nerve damage (12-mm gap) were divided into sham, autograft, PCL (empty NGC), and PCL + MSCs (NGC multi-functionalized with 106 canine AdMSCs embedded in heterologous fibrin biopolymer) groups. In vitro, the cells were characterized and directly stimulated with interferon-gamma to evaluate their neuroregeneration potential. In vivo, the sciatic and tibial functional indices were evaluated for 12 weeks. Gait analysis and nerve conduction velocity were analyzed after 8 and 12 weeks. Morphometric analysis was performed after 8 and 12 weeks following lesion development. Real-time PCR was performed to evaluate the neurotrophic factors BDNF, GDNF, and HGF, and the cytokine and IL-10. Immunohistochemical analysis for the p75NTR neurotrophic receptor, S100, and neurofilament was performed with the sciatic nerve. RESULTS: The inflammatory environment in vitro have increased the expression of neurotrophins BDNF, GDNF, HGF, and IL-10 in canine AdMSCs. Nerve guidance conduits multi-functionalized with canine AdMSCs embedded in HFB improved functional motor and electrophysiological recovery compared with PCL group after 12 weeks. However, the results were not significantly different than those obtained using autografts. These findings were associated with a shift in the regeneration process towards the formation of myelinated fibers. Increased immunostaining of BDNF, GDNF, and growth factor receptor p75NTR was associated with the upregulation of BDNF, GDNF, and HGF in the spinal cord of the PCL + MSCs group. A trend demonstrating higher reactivity of Schwann cells and axonal branching in the sciatic nerve was observed, and canine AdMSCs were engrafted at 30 days following repair. CONCLUSIONS: 3D-printed NGCs multi-functionalized with canine AdMSCs embedded in heterologous fibrin biopolymer as cell scaffold exerted neuroregenerative effects. Our multimodal approach supports the trophic microenvironment, resulting in a pro-regenerative state after critical sciatic nerve injury in rats.

Placenta-derived multipotent mesenchymal stromal cells: a promising potential cell-based therapy for canine inflammatory brain disease.

BACKGROUND: Canine inflammatory brain disease (IBD) is a severe inflammatory disorder characterized by infiltration of activated immune cell subsets into the brain and spinal cord. Multipotent mesenchymal stromal cells (MSCs) are a promising therapy for IBD, based on their potent pro-angiogenic, neuroprotective, and immunomodulatory properties. The aims of this study were to compare the immunomodulatory attributes of canine adipose-derived MSCs (ASCs) and placenta-derived MSCs (PMSCs) in vitro. These data will serve as potency information to help inform the optimal MSC cell source to treat naturally occurring canine IBD. METHODS: Indoleamine 2,3 dioxygenase (IDO) activity and prostaglandin E2 (PGE2) concentration at baseline and after stimulation with interferon gamma (IFNγ) and/or tumor necrosis factor alpha (TNFα) were measured from canine ASC and PMSC cultures. Leukocyte suppression assays (LSAs) were performed to compare the ability of ASCs and PMSCs to inhibit activated peripheral blood mononuclear cell (PBMC) proliferation. IDO activity and PGE2; interleukin (IL)-2, IL-6, and IL-8; TNFα; and vascular endothelial growth factor (VEGF) concentrations were also measured from co-culture supernatants. Cell cycle analysis was performed to determine how ASCs and PMSCs altered lymphocyte proliferation. RESULTS: Activated canine MSCs from both tissue sources secreted high concentrations of IDO and PGE2, after direct stimulation with IFNγ and TNFα, or indirect stimulation by activated PBMCs. Both ASCs and PMSCs inhibited activated PBMC proliferation in LSA assays; however, PMSCs inhibited PBMC proliferation significantly more than ASCs. Blocking PGE2 and IDO in LSA assays determined that PGE2 is important only for ASC inhibition of PBMC proliferation. Activated ASCs increased IL-6 and VEGF secretion and decreased TNFα secretion, while activated PMSCs increased IL-6, IL-8, and VEGF secretion. ASCs inhibited lymphocyte proliferation via cell cycle arrest in the G0/G1 and PMSCs inhibited lymphocyte proliferation via induction of lymphocyte apoptosis. CONCLUSION: Our results demonstrate that ASCs and PMSCs have substantial in vitro potential as a cell-based therapy for IBD; however, PMSCs more potently inhibited lymphocyte proliferation by inducing apoptosis of activated lymphocytes. These data suggest that the mechanism by which ASCs and PMSCs downregulate PBMC proliferation differs. Additional studies may elucidate additional mechanisms by which canine MSCs modulate neuroinflammatory responses.

Lissencephaly in Shih Tzu dogs.

BACKGROUND: Lissencephaly is a brain malformation characterized by smooth and thickened cerebral surface, which may result in structural epilepsy. Lissencephaly is not common in veterinary medicine. Here, we characterize the first cases of lissencephaly in four Shih Tzu dogs, including clinical presentations and findings of magnetic resonance imaging of lissencephaly and several concomitant brain malformations. CASE PRESENTATION: Early-onset acute signs of forebrain abnormalities were observed in all dogs, which were mainly cluster seizures and behavioral alterations. Based on neurological examination, the findings were consistent with symmetrical and bilateral forebrain lesions. Metabolic disorders and inflammatory diseases were excluded. Magnetic resonance imaging for three dogs showed diffuse neocortical agyria and thickened gray matter while one dog had mixed agyria and pachygyria. Other features, such as internal hydrocephalus, supracollicular fluid accumulation, and corpus callosum hypoplasia, were detected concomitantly. Antiepileptic drugs effectively controlled cluster seizures, however, sporadic isolated seizures and signs of forebrain abnormalities, such as behavioral alterations, central blindness, and strabismus persisted. CONCLUSIONS: Lissencephaly should be considered an important differential diagnosis in Shih Tzu dogs presenting with early-onset signs of forebrain abnormalities, including cluster seizures and behavioral alterations. Magnetic resonance imaging was appropriate for ante-mortem diagnosis of lissencephaly and associated cerebral anomalies.

Canine Adipose-Derived Mesenchymal Stromal Cells Enhance Neuroregeneration in a Rat Model of Sciatic Nerve Crush Injury.

Crush injuries in peripheral nerves are frequent and induce long-term disability with motor and sensory deficits. Due to axonal and myelin sheath disruptions, strategies for optimized axonal regeneration are needed. Multipotent mesenchymal stromal cells (MSC) are promising because of their anti-inflammatory properties and secretion of neurotrophins. The present study investigated the effect of canine adipose tissue MSC (Ad-MSC) transplantation in an experimental sciatic nerve crush injury. Wistar rats were divided into three groups: sham ( n = 8); Crush+PBS ( n = 8); Crush+MSC ( n = 8). Measurements of sciatic nerve functional index (SFI), muscle mass, and electromyography (EMG) were performed. Canine Ad-MSC showed mesodermal characteristics (CD34-, CD45-, CD44+, CD90+ and CD105+) and multipotentiality due to chondrogenic, adipogenic, and osteogenic differentiation. SFI during weeks 3 and 4 was significantly higher in the Crush+MSC group ( p < 0.001). During week 4, the EMG latency in the Crush+MSC groups had better near normality ( p < 0.05). The EMG amplitude showed results close to normality during week 4 in the Crush+MSC group ( p < 0.04). There were no statistical differences in muscle weight between the groups ( p > 0.05), but there was a tendency toward weight gain in the Crush+MSC groups. Better motor functional recovery after crush and perineural canine Ad-MSC transplantation was observed during week 2. This was maintained till week 4. In conclusion, the canine Ad-MSC transplantation showed early pro-regenerative effects between 2-4 weeks in the rat model of sciatic nerve crush injury.

Extradural lumbar nerve root cavernous hemangioma: case report and successful surgical resection / Hemangioma cavernoso extradural de raiz nervosa lombar: relato de caso e ressecção cirúrgica bem-sucedida

Extradural lumbar spinal canal cavernous hemangiomas (or cavernomas) are rare lesions that can induce intense back pain and neurological deficit. We present a case report of a patient with a pure radicular lombar extradural cavernoma resembling a benign neurological tumor in imaging exams and a successful surgical resection.

Os hemangiomas cavernosos do canal vertebral lombar extradural (ou cavernomas) são lesões raras que podem induzir dor intensa no dorso e déficit neurológico. Apresentamos um relato de caso de um paciente com um cavernoma extradural lombar radicular puro assemelhando-se a um tumor neurológico benigno em exames de imagem e uma ressecção cirúrgica bem-sucedida.

Safety and tracking of intrathecal allogeneic mesenchymal stem cell transplantation in healthy and diseased horses.

BACKGROUND: It is currently unknown if the intrathecal administration of a high dose of allogeneic mesenchymal stem cells (MSCs) is safe, how MSCs migrate throughout the vertebral canal after intrathecal administration, and whether MSCs are able to home to a site of injury. The aims of the study were: 1) to evaluate the safety of intrathecal injection of 100 million allogeneic adipose-derived MSCs (ASCs); 2) to assess the distribution of ASCs after atlanto-occipital (AO) and lumbosacral (LS) injection in healthy horses; and 3) to determine if ASCs homed to the site of injury in neurologically diseased horses. METHODS: Six healthy horses received 100 × 106 allogeneic ASCs via AO (n = 3) or LS injection (n = 3). For two of these horses, ASCs were radiolabeled with technetium and injected AO (n = 1) or LS (n = 1). Neurological examinations were performed daily, and blood and cerebrospinal fluid (CSF) were evaluated prior to and at 30 days after injection. Scintigraphic images were obtained immediately postinjection and at 30 mins, 1 h, 5 h, and 24 h after injection. Three horses with cervical vertebral compressive myelopathy (CVCM) received 100 × 106 allogeneic ASCs labeled with green fluorescent protein (GFP) via AO injection and were euthanized 1-2 weeks after injection for a full nervous system necropsy. CSF parameters were compared using a paired student's t test. RESULTS: There were no significant alterations in blood, CSF, or neurological examinations at any point after either AO or LS ASC injections into healthy horses. The radioactive signal could be identified all the way to the lumbar area after AO ASC injection. After LS injection, the signal extended caudally but only a minimal radioactive signal extended further cranially. GFP-labeled ASCs were not present at the site of disease at either 1 or 2 weeks following intrathecal administration. CONCLUSIONS: The intrathecal injection of allogeneic ASCs was safe and easy to perform in horses. The AO administration of ASCs resulted in better distribution within the entire subarachnoid space in healthy horses. ASCs could not be found after 7 or 15 days of injection at the site of injury in horses with CVCM.

Effects of Canine and Murine Mesenchymal Stromal Cell Transplantation on Peripheral Nerve Regeneration.

BACKGROUND AND OBJECTIVES: Maintaining a permissive microenvironment is essential for adequate nerve regeneration. Cell-based therapy has the potential based cell replacement and promotion of axonal growth. The adipose tissue derived mesenchymal stromal cells (Ad-MSC) attract interest because neuroregenerative and anti-inflammatory properties. The aim of this study was to evaluate the effects of canine and murine Ad-MSC transplantation on the sciatic nerve regeneration. METHODS: Forty Wistar rats were divided randomly into: control group - CG (n=8); denervated group - DG (n=8); decellularized vein group - VG (n=8); decellularized vein＋canine MSC-cMSC (n=8); descellularized vein＋murine MSC-mMSC (n=8). After 10-mm nerve gap, the tubulation technique was performed with decellularized vein filled with 106 MSC labeled with quantum dots (Qtracker 665®). The sciatic nerve functional index (SFI) and electroneuromyography (ENMG) measurements were carried and morphometric and immunohistochemistry analysis of the tissue. RESULTS: The SFI values were higher in the cMSC and mMSC groups at day 27 (p<0.020) and day 35 (p<0.011). The ENMG analysis also revealed better results in the mMSC group. Density, number, and total area of the fibers were increased in the mMSC and cMSC groups. Brain-derived neurotrophic factor BDNF and S-100 protein positive immunoreactivity showed a higher expression for both in the nerve of the mMSC and cMSC groups. The MSC labeled with quantum dots were detected at day 35, indicating neuronal survival long after the nerve damage. CONCLUSIONS: Murine and canine Ad-MSC associated with decellularized vein scaffold had positive effects on sciatic nerve regeneration in rats.

Prevalence of behavioral changes in senile dogs / Prevalência de alterações comportamentais em cães idosos

ABSTRACT: Owing to improvements in animal welfare, such as in nutrition, preventive medicine, and diagnostic tools, older dogs have become increasingly more prevalent than before. Unfortunately, there is an increasing number of dogs presenting behavioral changes correlated with this growth in the geriatric population. However, many of these owners consider these signs as normal for old age. This study aimed to determine the prevalence of clinical signs related with behavioral changes in the canine geriatric population admitted to the Veterinary Hospital of the Veterinary Medicine and Animal Science College of Universidade Estadual Paulista. The owners of 800 old dogs completed a questionnaire about disorientation, changes in social interaction, loss of training (house soiling), changes in the wake-sleep cycle, physical activity, memory, learning, consciousness, and perception. The results showed that the prevalence of behavioral changes in the canine geriatric population was 90.7% of dogs that presented at least one of these behavioral changes, and 22.3% with at least six changes, and with changes in physical activity (58.5%) and loss of acquired training (30.2%) being the highest percentage groups that were reported by the owners. We concluded that the prevalence of behavioral changes related with aging in the canine geriatric population was high, and that it was important to include questions about behavioral changes during regular veterinary clinical appointments to identify these clinical signs earlier and investigate the possible causes of the cognitive impairment, including neurological diseases, such as Canine Cognitive Dysfunction Syndrome.

RESUMO: Devido à melhora na nutrição e na medicina veterinária, há mais cães idosos do que antes. Infelizmente, acompanhando este crescimento na população geriátrica, houve um aumento no número de cães demonstrando distúrbios de comportamento e senilidade aparente. Entretanto, muitos dos proprietários têm considerado todos esses sinais decorrentes de uma inevitável velhice. O objetivo deste estudo foi obter a prevalência de sinais clínicos relacionados com alterações comportamentais na população canina idosa, atendida no Hospital Veterinário da Faculdade de Medicina Veterinária e Zootecnia da UNESP. Proprietários de 800 cães idosos responderam um questionário sobre desorientação, alterações na interação social, perda de treinamento (urina e/ou defeca em locais não habituais), alterações no ciclo sono-vigília, atividade física, memória, aprendizagem, consciência e percepção. Os resultados obtidos neste estudo apresentaram uma prevalência de alterações comportamentais na população canina idosa significativa, variando de 22,3% dos cães, com pelo menos seis alterações comportamentais e, chegando a 90,7% com pelo menos uma alteração de comportamento, sendo alterações na atividade física (58,5%) e perda de treinamentos adquiridos (30,2%) os grupos de maior percentual relatado pelos proprietários. Conclui-se que a prevalência de alterações comportamentais relacionadas com o envelhecimento da população canina é alta, sendo importante incluir questões sobre as alterações comportamentais durante as consultas clínicas veterinárias regulares para identificar, o mais precocemente possível, estes sinais clínicos e realizar ações para investigar as possíveis causas de déficit cognitivo, incluindo doenças neurológicas, como a Síndrome de Disfunção Cognitiva canina.

Disfagia devido a osteoitose cervical: relato de caso / Dysphagia due to cervical osteophytosis: case report

Doenças degenerativas na coluna cervical são comuns nos pacientes idosos. Os autores apresentam caso de paciente do sexo feminino, 44 anos, com quadro de disfagia progressiva relacionada à osteofitose cervical anterior em (C5-C6 e C6-C7). Esofagograma mostrou a compreensão esofágica pelos osteófitos anteriores. O tratamento realizado foi brocagem dos osteófitos, discectomia cervical em dois níveis e artrodese cervical, resultando na remissão completa do sintoma. Embora seja uma causa de disfagia, osteófito deve ser incluído como diagnóstico diferencial pois é uma causa tratável e reversível do sintoma.

Degenerative diseases of the cervical spine are common in elderly patients. The authors present a case report of female, 44 years old, with progressive dysphagia due to anterior cervical osteophytes (C5-C6 and C6-C7). Esophagogram showed esophageal compression by anterior osteophytes. The treatment was removal of osteophytes by drill, cervical discectomy on two levels and cervical arthrodesis, resulting in complete remission of symptoms. Although it is an un-common cause of dysphagia, osteophytes should be included in the differential diagnosis because it is a reatable and reversible cause of the symptom.

Feasibility and safety of intrathecal transplantation of autologous bone marrow mesenchymal stem cells in horses.

BACKGROUND: Recent studies have demonstrated numerous biological properties of mesenchymal stem cells and their potential application in treating complex diseases or injuries to tissues that have difficulty regenerating, such as those affecting the central and peripheral nervous system. Thus, therapies that use mesenchymal stem cells are promising because of their high capacity for self-regeneration, their low immunogenicity, and their paracrine, anti-inflammatory, immunomodulatory, anti-apoptotic and neuroprotective effects. In this context, the purpose of this study was to evaluate the feasibility and safety of intrathecal transplantation of bone marrow-derived mesenchymal stem cells in horses, for future application in the treatment of neurological diseases. RESULTS: During the neurological evaluations, no clinical signs were observed that were related to brain and/or spinal cord injury of the animals from the control group or the treated group. The hematological and cerebrospinal fluid results from day 1 and day 6 showed no significant differences (P > 0.05) between the treated group and the control group. Additionally, analysis of the expression of matrix metalloproteinase (MMP) -2 and -9 in the cerebrospinal fluid revealed only the presence of pro-MMP-2 (latent), with no significant difference (P > 0.05) between the studied groups. CONCLUSIONS: The results of the present study support the hypothesis of the feasibility and safety of intrathecal transplantation of autologous bone marrow-derived mesenchymal stem cells, indicating that it is a promising pathway for cell delivery for the treatment of neurological disorders in horses.

Traumatic arteriovenous fistula associated to pseudoaneurysm of the superficial temporal artery in a child: case report and literature review / Fístula arteriovenosa traumática associada a pseudoaneurisma da artéria temporal superficial em uma criança: relato de caso e revisão da literatura

As fístulas arteriovenosas (FAVs) e os pseudoaneurismas traumáticos extracranianos são malformações incomuns e, em sua maioria, estão associadas a traumatismo craniano fechado com lesão contusa de forte intensidade. O diagnóstico em geral é clínico, porém o exame de escolha para o diagnóstico definitivo é a angiografia. Nos casos em que a lesão é pequena, é possível abordá-la com embolização via endovascular com sucesso. A excisão cirúrgica, no entanto, ainda é o método de escolha para o tratamento. É relatado um caso de um paciente do sexo masculino, com 9 anos de idade, diagnosticado com FAV, acometendo o ramo frontal da artéria temporal superficial, secundária a trauma craniano contundente ocorrido três anos antes do diagnóstico.

The arteriovenous fistulae (AVFs) and the extracranial traumatic pseudoaneurysms are uncommon malformations and in the majority of the cases are associated to closed head trauma with high intensity blunt lesion. The diagnosis is generally clinical, though the exam of choice for definitive diagnosis is an angiography. In minor lesion cases it's possible to successfully approach it with endovas-cular embolization. The surgical excision though, is the method of choice for the treatment. Here is reported a case of a 9-year-old male patient, diagnosed with AVF involving the frontal branch of the superficial temporal artery, secondary to blunt head trauma occurred three years before diagnosis.

Equine mesenchymal stem cells from bone marrow, adipose tissue and umbilical cord: immunophenotypic characterization and differentiation potential.

INTRODUCTION: Studies with mesenchymal stem cells (MSCs) are increasing due to their immunomodulatory, anti-inflammatory and tissue regenerative properties. However, there is still no agreement about the best source of equine MSCs for a bank for allogeneic therapy. The aim of this study was to evaluate the cell culture and immunophenotypic characteristics and differentiation potential of equine MSCs from bone marrow (BM-MSCs), adipose tissue (AT-MSCs) and umbilical cord (UC-MSCs) under identical in vitro conditions, to compare these sources for research or an allogeneic therapy cell bank. METHODS: The BM-MSCs, AT-MSCs and UC-MSCs were cultured and evaluated in vitro for their osteogenic, adipogenic and chondrogenic differentiation potential. Additionally, MSCs were assessed for CD105, CD44, CD34, CD90 and MHC-II markers by flow cytometry, and MHC-II was also assessed by immunocytochemistry. To interpret the flow cytometry results, statistical analysis was performed using ANOVA. RESULTS: The harvesting and culturing procedures of BM-MSCs, AT-MSCs and UC-MSCs were feasible, with an average cell growth until the third passage of 25 days for BM-MSCs, 15 days for AT-MSCs and 26 days for UC-MSCs. MSCs from all sources were able to differentiate into osteogenic (after 10 days for BM-MSCs and AT-MSCs and 15 days for UC-MSCs), adipogenic (after 8 days for BM-MSCs and AT-MSCs and 15 days for UC-MSCs) and chondrogenic (after 21 days for BM-MSCs, AT-MSCs and UC-MSCs) lineages. MSCs showed high expression of CD105, CD44 and CD90 and low or negative expression of CD34 and MHC-II. The MHC-II was not detected by immunocytochemistry techniques in any of the MSCs studied. CONCLUSIONS: The BM, AT and UC are feasible sources for harvesting equine MSCs, and their immunophenotypic and multipotency characteristics attained minimal criteria for defining MSCs. Due to the low expression of MHC-II by MSCs, all of the sources could be used in clinical trials involving allogeneic therapy in horses. However, the BM-MSCs and AT-MSCs showed fastest ''in vitro'' differentiation and AT-MSCs showed highest cell growth until third passage. These findings suggest that BM and AT may be preferable for cell banking purposes.

Giant pericallosal artery aneurysm ? Literature review and report of a rare case / Aneurisma gigante de artéria pericalosa ? Revisão da literatura e relato de um caso raro

Giant pericallosal artery aneurysms are extremely rare. Aneurismatic lesions involving this artery are usually small, tend to early bleeding and might be associated with other lesions. Differential diagnosis of giant aneurysms are not easy and includes tumoral, infectious and vascular mass effect lesions. We report a case of a giant and partially thrombosed left pericallosal artery aneurysm. A 58-year-old man, presented with progressive headaches, seizures and speech alterations initially misdiagnosed as a falx cerebri meningioma. As clinical status continue to worsen, magnetic resonance imaging and digital cerebral angiography were performed and a vascular etiology was considered. The patient was then referred to our hospital for surgical treatment. The peculiarity of this case concerns the difficulty of surgical treatment once the surgeon was not able to obtain control of the afferent artery and the aneurysm neck could not be visualized. Also, the aneurysm adhered to the medial surfaces of the frontal lobes and covered the anterior cerebral arteries. Treatment by means of microsurgical thrombectomy, clipping and resection of the lesion was successfully performed. Microsurgical treatment may provide good results when carefully planned with the help of imaging studies of the lesion. It is essential to keep in mind that flexible approach is of great importance when dealing with giant aneurysmatic lesions of pericallosal artery due to its variety of intraoperative presentation.

Aneurismas gigantes da artéria pericalosa são raros. Lesões aneurismáticas acometendo essa artéria são, geralmente, pequenas, tendem a sangrar precocemente e associam-se a outras lesões.O diagnóstico diferencial dos aneurismas gigantes é complexo, uma vez que inclui diversas lesões que cursam com efeito de massa, entre elas, as tumorais, infecciosas e vasculares. Relatamos o caso de um aneurisma gigante, parcialmente trombosado, acometendo a artéria pericalosa esquerda. Tratase de um paciente de 58 anos de idade, sexo masculino, apresentando quadro de cefaleia, crises convulsivas e alterações na fala. O achado de uma massa inter-hemisférica de característica hiperdensaà tomografia computadorizada de crânio levou ao diagnóstico errôneo de meningioma da foice cerebral. Uma vez que o quadro clínico do paciente continuou a piorar, optou-se por submetê-lo à ressonância nuclear magnética e à angiografia cerebral digital. A hipótese de etiologia vascular foi então aventada. O paciente foi encaminhado ao nosso serviço para ser submetido a tratamento cirúrgico. A peculiaridade desse caso deve-se à dificuldade de acesso à artéria aferente e de visualização do colo para controle da lesão. Além disso, o aneurisma encontrava-se aderido às faces mediais dos lobos frontais e cobria as artérias cerebrais anteriores. O tratamento cirúrgico por meio de trombectomia microcirúrgica,clipagem e ressecção da lesão pôde ser realizado com sucesso. O tratamento microcirúrgico, quando cuidadosamente planejado por meio de exames de imagem, pode viabilizar a obtenção de resultadossatisfatórios. No entanto, deve-se atentar para o fato de que lesões aneurismáticas gigantes de artéria pericalosa podem se apresentar de diversas formas durante o ato operatório, sendo, portanto, essencialuma abordagem flexível, adaptada às peculiaridades de cada lesão.

Immunophenotypic, immunocytochemistry, ultrastructural, and cytogenetic characterization of mesenchymal stem cells from equine bone marrow.

The aim of this study was to isolate, culture, and characterize mesenchymal stem cells (MSCs) from horse bone marrow (BM) using the techniques of flow cytometry, immunocytochemistry, cytogenetics, and electron microscopy. Immunophenotypic analysis revealed the presence of MSCs with high expression of the CD90 marker, lower expression of the CD44 marker, and absent expression of the CD34 marker. In assays of differentiation, the positive response to osteogenic (OST), chondrogenic (CDG), and adipogenic (ADP) differentiation signals was observed and characterized by deposition of calcium-rich extracellular matrix (OST), proteoglycans and collagen II (CDG) and intracellular deposition of fat drops (ADP). In immunocytochemical characterization, MSCs were immunopositive for CD44, vimentin, and PCNA, and they were negative for CD13. In the ultrastructural analysis of MSCs, the most outstanding characteristic was the presence of rough endoplasmic reticulum with very dilated cisterns filled with a low electrodensity material. Additionally, MSCs had normal karyotypes (2n = 64) as evidenced by cytogenetic analysis, and aneuploidy in metaphase was not observed. The protocols for isolating, culturing, and characterizing equine MSCs used in this study were shown to be appropriate for the production of a cell population with a good potential for differentiation and without aneuploidy that can be used to study future cellular therapies.

Surto de abscesso mandibular por Pseudomonas aeruginosa em ovinos / Outbreak of mandibular abscess by Pseudomonas aeruginosa in sheep

Descreve-se um surto de abscesso mandibular em ovelhas da raça Bergamácia no município de Botucatu, estado de São Paulo. Do rebanho de 120 animais, 35 apresentaram aumento de volume mandibular com a presença de nódulos únicos, de consistência pétrea, de diferentes tamanhos, fistulados ou não e sem indicativos de inflamação dos tecidos moles adjacentes. Os animais eram criados em pasto de Panicum maximum cv. Tanzânia com água e sal mineral ad libitum e everminados, via oral, com pistolas dosificadoras. O material para diagnóstico microbiológico e antibiograma foi coletado de cinco animais acometidos, por punção e aspiração dos nódulos. Dos 35 animais acometidos, 19 foram submetidos ao exame radiográfico, um ao exame tomográfico e outro à biópsia óssea da região submandibular. O único ovino que morreu, encontrava-se em estado de caquexia provavelmente devido à localização do aumento de volume que afetou a implantação dos dentes molares daquela região impedindo a apreensão e mastigação adequadas levando a perda da condição corporal e morte. Ao exame necroscópico, observaram-se áreas de necrose caseosa na mandíbula direita de onde isolou-se Pseudomonas aeruginosa. O tratamento utilizado foi baseado na aplicação de iodeto de sódio a 10 por cento por via intramuscular e antibioticoterapia segundo antibiograma com enrofloxacina por via intramuscular, porém com pouca eficácia. Diante do quadro clínico, dos dados de anamnese, da localização das lesões no tecido ósseo mandibular, do resultado do cultivo microbiológico, das alterações radiográficas e tomográficas foi feito o diagnóstico de abscesso mandibular causado por Pseudomonas aeruginosa.

We describe an outbreak of mandibular abscess in Bergamácia sheep from the municipality of Botucatu, São Paulo, Brazil. In a herd of 120 sheep, 35 showed an increase of the mandibular volume with presence of a single nodule of stony consistency and different sizes, often fistulated but with no signs of inflammation in the surrounding soft tissues. The sheep were raised on a Panicum maximum cv. Tanzania pasture, with water and mineral salt ad libitum, and were treated orally against parasites with drenching pistols. Material for microbiological diagnosis and antibiogram was collected from the nodules of five affected sheep, through puncture and aspiration. From 35 affected sheep, 19 were submitted to radiographic examination, one was submitted to computer tomography and another to bone biopsy of the submandibular region. The only sheep that died was in a state of cachexia, probably due to the location of the volume increase which affected the molar hindering apprehension and appropriate chewing. Necropsy revealed areas of caseous necrosis in the jaw, from where Pseudomonas aeruginosa was isolated. The sheep were treated with intramuscular application of 10 percent sodium iodide and the antobiotgic enrofloxacin, but with little efficacy. Based on the clinical data from medical records, the location of lesions in the jaw bone, the result of microbiological culture, radiographic and computer tomography, the diagnosis of mandibular abscesses due to Pseudomonas aeruginosa infection was made.

Hematological and cerebrospinal fluid changes in cattle naturally and experimentally infected with the bovine herpesvirus 5

Bovine herpesvirus 5 (BoHV-5) meningoencephalitis is one of the main causes of mortality by encephalopathy in Brazilian cattle herds. However, the neurological signs observed are common to several encephalopathies and do not contribute decisively to a diagnosis. In order to verify hematological and cerebrospinal fluid (CSF) changes, blood and CSF samples from naturally and experimentally infected bovines were evaluated. In natural cases (n=17), the samples were collected only once, and in experimental cases (n=7), the samples were sequentially obtained throughout disease progression. While routine methods were used to examine the samples, BoHV-5 infection was confirmed by a PCR assay. Blood analyses did not reveal any consistent hematological alterations and the leukogram results occasionally showed increases in leukocyte and segmented neutrophil. Hyperfibrinogenemia was noted in all experimentally infected calves and in half of the natural cases. Pleocytosis with mononuclear cells was a remarkable finding in CSF collected from both groups of animals and was present even in experimentally infected calves that remained asymptomatic. Therefore, CSF evaluation can be used as an auxiliary method in ante-mortem BoHV-5 diagnosis.

A meningoencefalite determinada pelo herpesvírus bovino 5 (BoHV-5) é considerada uma das principais causas de mortalidade por encefalopatia em bovinos no Brasil. O diagnóstico clínico da infecção é difícil de ser realizado pois os sinais neurológicos ocasionados pela infecção com o BoHV-5 são comuns aos observados em encefalopatias bovinas de diferentes etiologias. Com o objetivo de determinar as alterações hematológicas e do líquido cefalorraquidiano, foram avaliadas amostras de sangue total e líquor colhidas de animais infectados tanto naturalmente quanto experimentalmente. Nos casos naturais da doença (n=17), as amostras foram coletadas apenas uma vez. Nos casos de infecção experimental (n=7), as amostras foram obtidas sequencialmente durante a evolução clínica da doença. Enquanto na análise das amostras clínicas foram empregadas metodologias rotineiras, a infecção pelo BoHV-5 foi confirmada em todos os animais incluídos nesse estudo por meio da técnica de PCR. As análises hematológicas não revelaram alterações consistentes e os resultados obtidos a partir do leucograma mostraram aumento ocasional de leucócitos e neutrófilos segmentados. A hiperfibrinogenemia pôde ser observada em todos os bezerros infectados experimentalmente e em metade dos casos naturais da doença. Diferentemente, a pleiocitose com predomínio de células mononucleares foi considerada um achado marcante nas amostras de líquor coletadas em ambos os grupos de animais, encontrando-se presente até mesmo nos bezerros infectados experimentalmente que permaneceram assintomáticos. Os resultados do presente estudo, obtidos tanto em condições de infecções naturais quanto experimentais, demonstraram que a avaliação do líquor cefalorraquidiano pode ser utilizada como método auxiliar para o diagnóstico ante-mortem da infecção pelo BoHV-5.

Mannheimiose pulmonar experimental em bezerros: swab nasal e nasofaringeano como auxílio diagnóstico: [revisão] / Experimental pneumonic mannheimiosis in calves: nasal and nasopharingeal swabs for diagnostic: [revision]

Um modelo experimental de mannheimiosepneumônica bovina (MPB) foi utilizado com o objetivo de avaliar as espécies bacterianas das cavidades nasais e nasofaringeanas em diferentes momentos do curso da doença, bem como verificar a eficiência diagnóstica do exame microbiológico dos swabs nasais (SN) e nasofaringeanos (SNF). Um total de 28 bezerros foi distribuído aleatoriamente em quatro grupos experimentais (G1 a G4). SN e SNF foram colhidos sete dias antes e 12 (G1), 24 (G2), 48 (G3) e 72 (G4) horas após a inoculação intrabronquial de Mannheimia haemolytica. Após a indução da MPB, a bactéria M. haemolytica biotipo A foi predominante nos SN e SNF, sendo isolada em todos os momentos avaliados, com exceção de um SN colhido 24 horas após a indução da infecção. Não houve diferença significativa nas taxas de isolamento de Pasteurella multocida nos SN ou SNF, colhidos antes e após a indução da MPB. Contudo, esta bactéria passou a ser isolada mais freqüentemente após a indução da MPB, principalmente no SNF. Portanto, pode-se concluir que o exame microbiológico de SN e SNF é um teste auxiliar no diagnóstico da MPB.

An experimental model of bovine pneumonic mannheimiosis (BPM) was used to evaluate the nasal and nasopharynx bacterial species of calves during the course of the disease and for checking the diagnostic efficiency of nasal swab (NS) and nasopharingeal swab (NPS) microbiological exams. A total of 28 calves were randomized into four experimental groups (G1-G4). NS and NPS were obtained 7 days before and 12 (G1), 24 (G2), 48 (G3) e 72 (G4) hours after intrabronchial inoculation of Mannheimia haemolytica. After the induction of BPM, M. haemolytica biotype A was the predominant isolated bacterium in NS and NPS in all evaluated sampling times, except for one NS (harvested 24 hours). There were no significant statistical differences for the rates of Pasteurella multocida isolation in NS and NPS, harvested before and after the induction of BPM. However, this bacterium was isolated more frequently after the induction of BPM, mainly in NPS. Therefore, the microbiological NS and NPS exams were important auxiliary tests for diagnosing BPM.