Comparison of three different strategies to treat sciatic nerve regeneration: an experimental study

Purpose: To compare the effect of vein conduit filled with adipose tissue stem cells (ASC) on peripheral nerve injury regeneration. Methods: We analyzed 30 male Wistar rats surgically submitted to a 5-mm gap on the sciatic nerve. Then, the animals were divided into three groups: nerve autografting (AG, n=10), autogenous inverted glycerol-conserved vein (VG, n=10), and autogenous inverted glycerol-conserved vein + ASC (VASCG, n=10). The study endpoints were neuromotor functional analysis, gastrocnemius muscle weight, and sciatic nerve graft histomorphometry analysis. In the histologic analysis, we added a control group (naïve nerve). Results: Regarding functional analysis (Walking tract- score), the findings at week 3 showed a difference between the AG and the VG (-96.6 vs. -59.6, p=0.01, respectively) and between the VG and the inverted vein + VASCG (-59.9 vs. -88.92, p=0.02). At week 12, this study showed a difference between the AG and the VG (-64.8 vs. -47.3, p=0.004, respectively), and also a difference between the VG and the VASCG (-47.3 vs. -57.4, p=0.02, respectively). There was no difference in the histomorphometry analysis (nerve diameter, Schwann cells counting). The gastrocnemius muscles on the intervention side were more atrophic when compared to the gastrocnemius muscles on the control side. Conclusions: Our results suggested better functional recovery in the inverted vein group when compared to control group, and inverted vein + ASC group.

Use of aerobic treadmill exercises on nerve regeneration after sciatic nerve injury in spontaneously hypertensive rats

Purpose: Various postoperative protocols have been proposed to improve outcomes and accelerate nerve regeneration. Recently, the use of physical exercise in a post-surgical neurorraphy procedure has shown good results when started early. We experimentally investigated the hypothesis that post-operative exercise speeds up results and improves clinical and morphologic parameters. Methods: Isogenic rats were randomly divided into four groups: 1 SHAM; 2 SHAM submitted to the exercise protocol (EP); 3 Grafting of the sciatic nerve; and 4 Grafting of the sciatic nerve associated with the EP. The EP was based on aerobic activities with a treadmill, with a progressive increase in time and intensity during 6 weeks. The results were evaluated by the sciatic functional index (SFI), morphometric and morphologic analysis of nerve distal to the lesion, and the number of spinal cord motor neurons, positive to the marker Fluoro-Gold (FG), captured retrogradely through neurorraphy. Results: Functional analysis (SFI) did not show a statistical difference between the group grafted with (­50.94) and without exercise (-65.79) after 90 days. The motoneurons count (Spinal cord histology) also showed no diference between these groups (834.5 × 833 respectively). Although functionally there is no difference between these groups, morphometric study showed a greater density (53.62) and larger fibers (7.762) in GRAFT group. When comparing both operated groups with both SHAM groups, all values were much lower. Conclusions: The experimental model that this aerobic treadmill exercises protocol did not modify nerve regeneration after sciatic nerve injury and repair with nerve graft.

Study on the mechanisms of Suaeda rigida polysaccharides on the heart inhibition and skeletal muscle promotion in the frog / [Estudo sobre os mecanismos dos polissacarídeos de Suaeda rigida na inibição do coração e promoção da musculatura esquelética no sapo]

The objectives of this study were to investigate the effects of different concentrations of Suaeda rigida polysaccharides (SRPs) on the physiological characteristics of the frog heart and gastrocnemius muscle, compare their similarities and differences, and analyze the mechanisms. CaCl2 and acetylcholine (Ach) were selected respectively to be co-incubated with the high concentration SRPs to observe the effects on the heart contraction of frog. The effects of different concentrations of the SRPs on the activities of acetylcholinesterase (A-CHE), Na+-K+-ATPase and Ca2+-Mg2+-ATPase in the isolated frog heart were detected by UV-Vis spectrophotometry. The gastrocnemius muscle was immersed in the high concentration of SRPs for 10 min, and the systolic indexes were recorded. The effects of SRPs on the Ach content and A-CHE activity at the sciatic nerve-gastrocnemius junction were determined by UV-Vis spectrophotometry and enzyme-linked immunosorbent assay (ELISA). The results showed that the SRPs had significant inhibitory effects on the contractile amplitude of isolated heart and the contractile amplitude induced by CaCl2 and Ach, respectively (P<0.01). The activity of Ca2+-Mg2+-ATPase was significantly promoted, and the activity of A-CHE was significantly inhibited (P<0.01). The contraction amplitude, contraction rate, relaxation rate of gastrocnemius muscle and the Ach content at the junction of sciatic nerve-gastrocnemius muscle were significantly increased (P<0.01), and the activity of A-CHE at the junction was significantly inhibited (P<0.01) by the SRPs. All the results suggested that the SRPs could inhibit the contraction of heart and promote the contraction and relaxation of skeletal muscle. The mechanism was related to blocking the fast INa channel, inhibiting the ICa-L and activating the M receptors of myocardial membrane and then inhibiting external Ca2+ influx, increasing Ca2+-Mg2+-ATPase activity, decreasing a-che activity.

Os objetivos deste estudo foram investigar os efeitos de diferentes concentrações de Suaeda rigida polissacarídeos (SRPs) sobre as características fisiológicas do coração de rã e do músculo gastrocnêmio, comparar suas semelhanças e diferenças, e analisar os mecanismos. CaCl2 e acetilcolina (Ach) foram selecionados respectivamente para serem co-incubados com os SRPs de alta concentração para observar os efeitos sobre a contração do coração de rã. Os efeitos das diferentes concentrações dos SRPs sobre as atividades da acetilcolinesterase (A-CHE), Na+-K+-ATPase e Ca2+-Mg2+-ATPase no coração isolado de rã foram detectados pela espectrofotometria UV-Vis. O músculo gastrocnêmio foi imerso na alta concentração de SRP por 10 min, e os índices sistólicos foram registrados. Os efeitos das SRPs no conteúdo de Ach e na atividade de A-CHE na junção nervo-gastrocnêmio ciático foram determinados pela espectrofotometria UV-Vis e pelo ensaio de imunoabsorção enzimática (ELISA). Os resultados mostraram que os SRPs tiveram efeitos inibidores significativos sobre a amplitude contrátil do coração isolado e a amplitude contrátil induzida por CaCl2 e Ach, respectivamente (P<0,01). A atividade do Ca2+-Mg2+-ATPase foi significativamente promovida e a atividade do A-CHE foi significativamente inibida (P<0,01). A amplitude de contração, a taxa de contração, a taxa de relaxamento do músculo gastrocnêmio e o conteúdo de Ach na junção do músculo nervo ciático-gastrocnêmio foram significativamente aumentados (P<0,01), e a atividade do A-CHE na junção foi significativamente inibida (P<0,01) pelas SRPs. Todos os resultados sugeriram que os SRPs poderiam inibir a contração do coração e promover a contração e o relaxamento do músculo esquelético. O mecanismo estava relacionado ao bloqueio do canal INa rápido, inibindo a ICa-L e ativando os receptores M da membrana miocárdica e depois inibindo o influxo externo de Ca2+, aumentando a atividade de Ca2+-Mg2+-ATPase, diminuindo a atividade a-che.

Marek's disease in roosters in the Potiguar semi-arid region

Background: Marek's disease (MD) is a transmissible disease in chickens caused by Gallid alphaherpesvirus 2 (GaHV-2). The infection is characterized by lymphocyte cellular infiltrates in peripheral nerves and other organs and tissues, including the skin; which can lead to dysfunction causing progressive asymmetric paresis and complete spastic paralysis of body extremities. Dermatitis and cardiac myositis caused by GaHV-2 in free-range chickens has rarely been described in Brazil. This reports the occurrence of the disease with a confirmatory molecular diagnosis in free-range poultry showing signs of dermatitis, poor performance, and cachexia and no mortality in the semi-arid Potiguar region. Cases: Twenty roosters of the Shamo lineage, among a brood of 42 birds, had a history of progressive weight loss and skin lesions. Two birds with poor body condition, erythema, and scaling of the skin in the head and cervical regions were sent for clinical care. All birds were between 12 and 18 months of age and were vaccinated against Newcastle disease and Fowlpox with only a few receiving vaccines against MD and Gumboro disease. According to the owner's report, some birds were previously kept outdoors, and when they were transferred to a small shed with little air circulation, they began to develop clinical signs after approximately 15 days. The first signs of the disease were also reported to have appeared 2.5 months before clinical care and, in the meantime, several treatments were instituted without success. Owing to the general condition of the animals and inconclusive clinical suspicion, the birds were subjected to euthanasia and necropsy. Tissue samples were collected for histopathological and polymerase chain reaction analyses to search for the GaHV-2 DNA meq gene. The main clinicopathological findings were erythema (47%, 20/42) and desquamation of skin and mild, prominent white multifocal areas in the heart. Histopathology revealed infiltration of pleomorphic lymphoblastic cells in the skin, heart, and sciatic nerve. The amplification of the L-meq and meq oncoprotein genes in these organs and in the liver, confirmed the infection by GaHV-2, consistent with that of a field strain. Discussion: MD was confirmed based on the macroscopic and histological lesions, and with the detection of GaHV-2 DNA in the affected tissues. The unusual clinical presentation represented an initial challenge for diagnosis. The clinical history was important to lead to the suspicion of MD, as roosters initiated clinical signs 15 days after they were transferred to a small shed with poor air circulation. This probably favored the high viral concentration and disease transmission among susceptible birds in the brood because the feather follicle is the primary site of viral replication for transmission; and desquamation of infected epithelial cells favor airborne horizontal transmission to susceptible chickens. The roosters had not been vaccinated against MD, which probably favored the infection, as vaccination is known to be a fundamental approach for MD control for effective growth of the poultry industry. Clinical findings and lesions, together with viral molecular detection, were fundamental for the diagnosis, a premise for the application of adequate prevention and control measures for the disease in breeding. This is the first report of MD with a confirmatory molecular diagnosis in northeastern Brazil.

Utilização da fisioterapia na reabilitação de bezerro com poliartrite e paresia neuromuscular: relato de caso / Use of physiotherapy in the rehabilitation of calves with polyarthritis and neuromuscular paresis: case report / Uso de la fisioterapia en la rehabilitación de un ternero con poliartritis y parisia neuromuscular: reporte de caso

A reabilitação animal na medicina veterinária é um campo recente, porém crescente a cada dia. Neste sentido, a fisioterapia veterinária atua trazendo inúmeros benefícios, como melhoria dos movimentos, redução da dor, edema e outras. Interfere ainda no tempo de recuperação, redução de custos para o proprietário, podendo ser hoje utilizada como um tratamento na recuperação pós-cirúrgica. Desta forma, objetivou-se relatar o uso da fisioterapia em uma bezerra com poliartrite e paresia neuromuscular. O animal apresentou um aumento de volume nas articulações cárpicas e társicas após histórico de onfalite, permanecendo em decúbito esternal por vários dias. O proprietário resolveu aplicar ferro dextrano na região glútea do animal, o que causou uma lesão de nervo isquiático. Foram instituídos protocolos medicamentosos e fisioterápicos, que culminaram no estímulo da marcha, propriocepção, além de hipertrofia muscular. Porém, em virtude de complicações resultantes de onfalite, o animal veio a óbito.(AU)

Animal rehabilitation in veterinary medicine is a recent field, but growing every day. In this sense, veterinary physiotherapy works bringing numerous benefits, such as improved movements, reduced pain, edema and others. Reducing the recovery time, reducing costs for the owner, and today it can be used as a treatment in post-surgical recovery. In this sense, the objective was to report the use of physiotherapy in a heifer with polyarthritis and neuromuscular paresis. The animal showed an increase in volume in the carpal and tarsal joints after a history of omphalitis, remaining in sternal decubitus for several days. The owner decided to apply iron dextran to the animal's gluteal region, which caused an injury to the sciatic nerve. Medicinal and physical therapy protocols were instituted, which culminated in the stimulation of gait, proprioception, in addition to muscle hypertrophy. However, due to complications resulting from omphalitis, the animal died.(AU)

La rehabilitación animal en medicina veterinaria es un campo reciente, pero en crecimiento cada día. En este sentido, la fisioterapia veterinaria actúa aportando numerosos beneficios, como mejora de los movimientos, reducción del dolor, edemas y otros. Disminución del tiempo de recuperación, reducción de costos para el propietario, pudiendo ser utilizado hoy en día como tratamiento en la recuperación posquirúrgica. En ese sentido, el objetivo fue reportar el uso de fisioterapia en una vaquilla con poliartritis y paresia neuromuscular. El animal presentó un aumento de volumen en las articulaciones del carpo y del tarso tras un antecedente de onfalitis, permaneciendo en decúbito esternal durante varios días. El propietario decidió aplicar hierro dextrano en la región glútea del animal, lo que provocó una lesión en el nervio ciático. Se instauraron protocolos farmacológicos y de fisioterapia, que culminaron con estimulación de la marcha, propiocepción, además de hipertrofia muscular. Sin embargo, debido a complicaciones derivadas de la onfalitis, el animal falleció.(AU)

Comparison of two volumes for sciatic perineural injection using 1% methylene blue in rabbit cadavers / [Comparação de dois volumes para injeção perineural no nervo isquiático utilizando azul de metileno 1% em cadáveres de coelhos]

The purpose of this study was to assess the efficacy of a blind technique for sciatic and femoral nerve block in rabbit cadavers by evaluating the spread of 1% methylene blue at two different volumes. Nine recently euthanized rabbits weighing 2.5(0.3kg were used. The sciatic (SN) and femoral (FN) nerves of each limb were randomly assigned for injection with 1% methylene blue at 0.2mL/kg (G0.2) or 0.3mL/kg (G0.3). Nerves were dissected and measured for depth and extension of staining (cm). Mean comparisons were performed using paired t test. The relation between volume and nerve staining ( 2cm was assessed using chi-square test. The mean depth of SN was 1.9±0.2 and 1.6±0.3cm and staining 1.9±1.4 and 2.0±1.2cm, respectively in G0.2 and G0.3. No relation was found between depth and dye spread and there was no association between nerve staining ( 2.0cm and volume of solution. The FN failed to be stained in all subjects. In conclusion, SN injection can be successfully performed without guidance in rabbits. The lower volume (0.2mL/kg) is recommended to avoid systemic toxicity.(AU)

O objetivo deste estudo foi avaliar a eficácia de uma técnica para bloqueio às cegas dos nervos isquiático e femoral em cadáveres de coelhos, por meio da avaliação da dispersão de azul de metileno 1% em dois volumes distintos. Nove coelhos recém-eutanasiados, com peso 2,5(0,3kg, foram utilizados. Os nervos isquiático (NI) e femoral (NF) de cada membro foram aleatoriamente designados para injeção com azul de metileno 1% a 0,2mL/kg (G0,2) ou 0,3mL/kg (G0,3). Em seguida, foram dissecados e mensurados em relação à sua profundidade e extensão corada (cm). As médias foram comparadas por meio de teste t pareado. A relação entre volume e extensão corada ( 2cm foi avaliada utilizando-se teste de qui-quadrado. A profundidade média do NI foi 1,9±0,2 e 1,6±0,3cm, e a extensão corada 1,9±1,4 e 2,0±1,2cm, respectivamente, no G0,2 e no G0,3. Não houve relação entre a profundidade e a extensão corada ou entre a extensão corada ( 2,0cm e o volume de solução. Não foi observada coloração do NF em nenhum cadáver. Concluiu-se que a injeção do NI pode ser realizada com sucesso sem auxílio de tecnologias em coelhos. O menor volume (0,2mL/kg) é recomendado para evitar toxicidade sistêmica.(AU)

Hypersensitivity in chicken (Gallus gallus domesticus) due to the association of lidocaine and bupivacaine in neural-guided femoral and sciatic nerve block

Background: Osteosyntheses, orthopedic surgeries that cause highly painful stimulation, are increasingly common inveterinary medicine. Epidural anesthesia is used to provide intraoperative and postoperative analgesia in mammals undergoing pelvic limb surgery. In birds, the synsacrum, the bone originating from the fusion of the lumbar and sacral vertebrae,makes this route inapplicable, thus peripheral nerve block is an easier option in this species. This report describes a caseof local hypersensitivity following the association of lidocaine and bupivacaine in anesthetic blocks of the femoral andsciatic nerves in chickens (Gallus gallus domesticus).Case: A study was conducted in chickens evaluating the effectiveness of anesthetic sciatic and femoral nerve blocks, guidedby a neural stimulator. Thirty-two 42-day-old male chickens of the species Gallus gallus domesticus, double breasted,weighing 1.86 ± 0.5 kg, were randomly divided into four groups: control (CG), lidocaine (LG), bupivacaine (BC) andthe association of lidocaine and bupivacaine (LBG). The doses used were 4 mg/kg of 2% lidocaine and 2 mg/kg of 0.5%bupivacaine, without vasoconstrictor. For CG, 0.9% NaCl solution was used, respecting the total volume of 1 mL/kg.Only one bird from the LBG showed side effects, presenting sensory and motor loss for 24 h after the administration ofthese drugs, before euthanasia was performed using anesthetic induction with isoflurane through a face mask, followedby the intravenous administration of propofol and then potassium chloride. The chicken was submitted to a necropsy andmacroscopically, soft, irregular, brownish lesions with a grayish focus were observed...

Hypersensitivity in chicken (Gallus gallus domesticus) due to the association of lidocaine and bupivacaine in neural-guided femoral and sciatic nerve block

Background: Osteosyntheses, orthopedic surgeries that cause highly painful stimulation, are increasingly common inveterinary medicine. Epidural anesthesia is used to provide intraoperative and postoperative analgesia in mammals undergoing pelvic limb surgery. In birds, the synsacrum, the bone originating from the fusion of the lumbar and sacral vertebrae,makes this route inapplicable, thus peripheral nerve block is an easier option in this species. This report describes a caseof local hypersensitivity following the association of lidocaine and bupivacaine in anesthetic blocks of the femoral andsciatic nerves in chickens (Gallus gallus domesticus).Case: A study was conducted in chickens evaluating the effectiveness of anesthetic sciatic and femoral nerve blocks, guidedby a neural stimulator. Thirty-two 42-day-old male chickens of the species Gallus gallus domesticus, double breasted,weighing 1.86 ± 0.5 kg, were randomly divided into four groups: control (CG), lidocaine (LG), bupivacaine (BC) andthe association of lidocaine and bupivacaine (LBG). The doses used were 4 mg/kg of 2% lidocaine and 2 mg/kg of 0.5%bupivacaine, without vasoconstrictor. For CG, 0.9% NaCl solution was used, respecting the total volume of 1 mL/kg.Only one bird from the LBG showed side effects, presenting sensory and motor loss for 24 h after the administration ofthese drugs, before euthanasia was performed using anesthetic induction with isoflurane through a face mask, followedby the intravenous administration of propofol and then potassium chloride. The chicken was submitted to a necropsy andmacroscopically, soft, irregular, brownish lesions with a grayish focus were observed...(AU)

Characterization of the first two toxins isolated from the venom of the ancient scorpion Tityus (Archaeotityus) mattogrossensis (Borelli, 1901)

Abstract Background: Almost all Tityus characterized toxins are from subgenera Atreus and Tityus, there are only a few data about toxins produced by Archaeotityus, an ancient group in Tityus genus. Methods: Tityus (Archaeotityus) mattogrossensis crude venom was fractionated by high performance liquid chromatography, the major fractions were tested in a frog sciatic nerve single sucrose-gap technique. Two fractions (Tm1 and Tm2) were isolated, partially sequenced by MALDI-TOF/MS and electrophysiological assayed on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells. Results: The sucrose-gap technique showed neurotoxicity in four fractions. One fraction caused a delay of action potential repolarization and other three caused a reduction in amplitude. An electrophysiological assay showed that Tm1 is active on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells, and Tm2 on HEK293 Nav 1.3 and DRG cells, but not in HEK293 Nav 1.6. In addition, Tm1 and Tm2 did promote a shift to more negative potentials strongly suggesting that both are -NaScTx. Conclusion: Although Tityus (Archaeotityus) mattogrossensis is considered an ancient group in Tityus genus, the primary structure of Tm1 and Tm2 is more related to Tityus subgenus. The patch clamp electrophysiological tests suggest that Tm1 and Tm2 are NaScTx, and also promoted no shift to more negative potentials, strongly suggesting that both are -NaScTx. This paper aimed to explore and characterize for the first time toxins from the ancient scorpion Tityus (Archaeotityus) mattogrossensis.

Characterization of the first two toxins isolated from the venom of the ancient scorpion Tityus (Archaeotityus) mattogrossensis (Borelli, 1901)

Background: Almost all Tityus characterized toxins are from subgenera Atreus and Tityus, there are only a few data about toxins produced by Archaeotityus, an ancient group in Tityus genus. Methods: Tityus (Archaeotityus) mattogrossensis crude venom was fractionated by high performance liquid chromatography, the major fractions were tested in a frog sciatic nerve single sucrose-gap technique. Two fractions (Tm1 and Tm2) were isolated, partially sequenced by MALDI-TOF/MS and electrophysiological assayed on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells. Results: The sucrose-gap technique showed neurotoxicity in four fractions. One fraction caused a delay of action potential repolarization and other three caused a reduction in amplitude. An electrophysiological assay showed that Tm1 is active on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells, and Tm2 on HEK293 Nav 1.3 and DRG cells, but not in HEK293 Nav 1.6. In addition, Tm1 and Tm2 did promote a shift to more negative potentials strongly suggesting that both are α-NaScTx. Conclusion: Although Tityus (Archaeotityus) mattogrossensis is considered an ancient group in Tityus genus, the primary structure of Tm1 and Tm2 is more related to Tityus subgenus. The patch clamp electrophysiological tests suggest that Tm1 and Tm2 are NaScTx, and also promoted no shift to more negative potentials, strongly suggesting that both are α-NaScTx. This paper aimed to explore and characterize for the first time toxins from the ancient scorpion Tityus (Archaeotityus) mattogrossensis.(AU)