PD-L1 overexpression induces STAT signaling and promotes the secretion of pro-angiogenic cytokines in non-small cell lung cancer (NSCLC).

BACKGROUND: Monoclonal antibodies (ICI) targeting the immune checkpoint PD-1/PD-L1 alone or in combination with chemotherapy have demonstrated relevant benefits and established new standards of care in first-line treatment for advanced non-oncogene addicted non-small cell lung cancer (NSCLC). However, a relevant percentage of NSCLC patients, even with high PD-L1 expression, did not respond to ICI, highlighting the presence of intracellular resistance mechanisms that could be dependent on high PD-L1 levels. The intracellular signaling induced by PD-L1 in tumor cells and their correlation with angiogenic signaling pathways are not yet fully elucidated. METHODS: The intrinsic role of PD-L1 was initially checked in two PD-L1 overexpressing NSCLC cells by transcriptome profile and kinase array. The correlation of PD-L1 with VEGF, PECAM-1, and angiogenesis was evaluated in a cohort of advanced NSCLC patients. The secreted cytokines involved in tumor angiogenesis were assessed by Luminex assay and their effect on Huvec migration by a non-contact co-culture system. RESULTS: PD-L1 overexpressing cells modulated pathways involved in tumor inflammation and JAK-STAT signaling. In NSCLC patients, PD-L1 expression was correlated with high tumor intra-vasculature. When challenged with PBMC, PD-L1 overexpressing cells produced higher levels of pro-angiogenic factors compared to parental cells, as a consequence of STAT signaling activation. This increased production of cytokines involved in tumor angiogenesis largely stimulated Huvec migration. Finally, the addition of the anti-antiangiogenic agent nintedanib significantly reduced the spread of Huvec cells when exposed to high levels of pro-angiogenic factors. CONCLUSIONS: In this study, we reported that high PD-L1 modulates STAT signaling in the presence of PBMC and induces pro-angiogenic factor secretion. This could enforce the role of PD-L1 as a crucial regulator of the tumor microenvironment stimulating tumor progression, both as an inhibitor of T-cell activity and as a promoter of tumor angiogenesis.

Fast liquid chromatography-tandem mass spectrometry method for the simultaneous determination of phytocannabinoids in oily based preparations.

The reported method aims to be a powerful aid for the simultaneous determination of tetrahydrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN), cannabigerol (CBG), tetrahydrocannabinolic acid (THCA), cannabidiolic acid (CBDA), and tetrahydrocannabivarin (THCV) in oily based preparations. The chromatographic separation was carried out using an Hypersil Gold PFP (50 × 2.1 mm, 1.9 µm) column, using H2O + 2 mM ammonium formate + 0.2 % formic acid (M1) and Methanol + 2 mM ammonium formate + 0.2 % formic acid (M2) as mobile phases. The flow rate was set 0.4 mL/min. Specifically, this method was validated in terms of linearity, limit of detections and quantifications (LODs and LOQs), accuracy (precision and trueness, both intra and interday), selectivity, and matrix effects. This procedure allowed quantifying seven phytocannabinoids in less than 10 min. The validated method shows a good linearity within the range 0.25-1000 ng/mL, while precision and trueness (intra- and inter-day) were below <13.25 % and 7.59 %, respectively. Regarding the matrix effect, the method satisfies all the requirements, except for the THC and THCV, where it reaches about 120 %. This element does not affect the method performances as it has been observed that this value is constant and reproducible and therefore does not involve errors in the quantitative analysis. The method was tested and applied on more 70 different oily based preparations. Furthermore, starting from four different cannabis cultivar (FM2, Bedrolite, Bedrocan, and Bediol), it allowed to evaluate the reproducibility of the magistrali preparations. The real samples, in fact, derive from different local pharmacies, and were analyzed by the accredited UNI CEI EN ISO/IEC 17025:2018, Pharmatoxicology Laboratory (ACCREDIA, lab n. 2274 ASLPE, accreditation number 1822 L), accordingly to the current regulations.

Hepatopatia esteroidal em gatas após terapia com prednisolona: aspectos laboratoriais, tomográficos e histopatológicos / [Steroid hepatopathy in cats after prednisolone use: laboratory, tomographic, and histopathologic aspects]

Glicocorticoides são amplamente utilizados na clínica de pequenos animais, entretanto seu uso contínuo pode causar efeitos colaterais. Os gatos são considerados menos susceptíveis a esses efeitos do que outras espécies, mas existem poucos trabalhos abordando os efeitos adversos em felinos. O objetivo deste estudo foi avaliar possíveis alterações laboratoriais, histopatológicas e do grau de atenuação radiográfica do parênquima hepático de gatas submetidas à terapia com prednisolona. Um ensaio clínico foi realizado em quatro gatas hígidas, as quais receberam prednisolona, por via oral, na dose de 3mg/kg, durante 60 dias consecutivos. Nos achados histopatológicos após 60 dias de tratamento, observou-se desorganização dos cordões de hepatócitos e degeneração vacuolar, além de necrose de hepatócitos, porém não foram observados sinais de fibrose no parênquima hepático. Os dados da tomografia computadorizada demonstram aumento do grau de atenuação do parênquima hepático a partir do 30º dia da administração de prednisolona, que persistiu até o final do experimento. No presente estudo, foi possível caracterizar a existência de hepatopatia esteroidal em gatos em estágios precoces da terapia com prednisolona.(AU)

Glucocorticoids are widely used medications in small animal practice; however, its continuous use can have side effects. Cats are considered less susceptible than other species, however, the literature does not usually address adverse effects in felines. The objective of this study was to evaluate possible laboratory and histopathologic changes, as well as changes to the degree of radiographic attenuation of the hepatic parenchyma in cats treated with prednisolone. A clinical trial was done in four healthy cats, who received prednisolone orally at 3mg/kg during 60 consecutive days. In the histopathologic findings at 60 days of treatment, there were disorganized hepatocyte chords and vacuolar degeneration, as well as hepatocyte necrosis, however, there were no signs of fibrosis in the hepatic parenchyma. Data obtained via computed tomography showed increase of the degree of attenuation in the hepatic parenchyma from day 30 of prednisolone therapy, which persisted until the end of the experiment. In the present study, it was possible to characterize the existence of steroidal hepathopathy in cats in the early stages of prednisolone therapy.(AU)

Acute-on-chronic subdural cranial hematoma in a dog: case report / Hematoma craniano subdural crônico-agudizado em um cão: relato de caso

A ten-month-old male Yorkshire terrier dog was evaluated via CT four months after traumatic brain injury. The head CT scan showed dilated ventricles associated with a peripheral crescent-shaped collection of blood near the right parietal bone with a mineralized area. The visceral layer of the hematoma was hyperattenuating on the native scan and showed moderate contrast enhancement after administration of intravenous iodinated contrast medium. No fractures were identified in the calvarium. These findings were compatible with acute-on-chronic calcified subdural hematoma, which have been described with more detail in humans. This is the first report to include the description of the imaging findings related to this condition using computed tomography in a dog.(AU)

Um cão Yorkshire Terrier, de 10 meses de idade, foi avaliado por tomografia computadorizada, quatro meses após lesão encefálica de origem traumática. A tomografia da cabeça demonstrou dilatação dos ventrículos associada com coleção de sangue periférico, em formato crescente, próxima ao osso parietal direito, com uma área mineralizada interna. A camada visceral do hematoma era hiperatenuante no exame pré-contraste e demonstrou moderado aumento de atenuação radiográfica após a administração de contraste iodado intravenoso. Fraturas no calvário não foram identificadas. Esses achados foram compatíveis com agudização de hematoma subdural crônico calcificado, condição que tem sido descrita com mais detalhes em humanos. Este é o primeiro relato que descreve os achados de imagem, por tomografia computadorizada, em um cão, relacionados a essa condição.(AU)

Cranial cavity filled with cerebrospinal fluid in an otter (Lontra longicaudis) - case report / Cavitação craniana preenchida por fluido cerebroespinhal em lontra (Lontra longicaudis) - relato de caso

Computed tomography of the head of an otter with a history of incoordination, visual deficits, and seizures was performed. Intracranial images revealed a large non-enhancing fluid attenuating cystic lesion in the left frontoparietal region communicating with the left lateral ventricle and subarachnoid space. These findings are consistent with a congenital brain cavity filled by cerebrospinal fluid, with porencephaly being the most probable diagnosis based on the clinical and tomographic findings. The authors highlight the rarity of such a cystic lesion in wild animals, with this being the first report in otters.(AU)

Realizou-se o exame tomográfico do crânio de uma lontra com histórico de incoordenação, déficit visual e convulsões. Imagens intracranianas revelaram grande lesão cística não captante de contraste em região frontoparietal esquerda, apresentando comunicação com o ventrículo lateral esquerdo e espaço subaracnoidea. Esses achados foram consistentes com cavitação cerebral congênita preenchida por fluido cerebroespinhal, sendo porencefalia o diagnóstico de maior probabilidade com base nos achados clínicos e tomográficos. Destaca-se a raridade dessa condição em animais silvestres, sendo este o primeiro relato em lontras.(AU)

Consequences of intraoperative spinal cord manipulation in dogs with thoracolumbar intervertebral disc extrusion / Consequências da manipulação medular intraoperatória em cães com extrusão intervertebral de disco toracolombar

The objective of the present study was to evaluate if extradural contact during hemilaminectomy would cause neurological deterioration in the early and/or late postoperative period in dogs with intervertebral disc extrusion. Nineteen dogs with thoracolumbar intervertebral disc extrusion underwent hemilaminectomy for spinal cord decompression and removal of extruded disc material. Meningeal contacts during surgery were quantified. Paraplegia (with nociception) and paraparesis were observed in 11/19 and 8/19 of dogs, respectively, before surgery. At the end of our study, only two (2/19) had paraplegia and one (1/19), paraparesis. There were more extradural contacts when extruded intervertebral disc material was at a ventrolateral position. Extradural contacts during surgery had no influence on neurological progression nor on time to recovery of motor function. Immediately (24 and 48 hours) after surgery, 13/19 dogs had the same neurological stage before surgery. At 7 and 90 days, 13/19 and 17/19 dogs, respectively, showed neurological improvement, compared with their preoperative stage. There was no influence of the number of extradural contacts on neurological recovery. These findings indicate that a careful inspection of the vertebral canal for removal of as much extruded disc material as possible does not cause neurologic deterioration.(AU)

O presente trabalho teve como objetivo avaliar se contatos extradurais durante hemilaminectomia em cães com extrusão de disco intervertebral causariam piora neurológica no pós-operatório imadiato e/ou tardio. Dezenove cães com extrusão toracolombar de disco intervertebral foram submetidos à hemilaminectomia para descompressão medular e remoção do material extruso. Durante o procedimento cirúrgico, os contatos meningomedulares foram quantificados. Antes da cirurgia, 11/19 cães apresentavam paraplegia (com nocicepção) e 8/19 cães, paraparesia. Ao fim do estudo, apenas dois cães (2/19) mostravam paraplegia com dor profunda e um (1/19), paraparesia. Observou-se maior quantidade de contatos extradurais quando o material discal extruso encontrava-se em posição ventrolateral. Os contatos extradurais não mostraram influência estatística na evolução neurológica dos animais, bem como no tempo de recuperação das funções motora. Vinte e quatro e 48 horas após a cirurgia, 13/19 cães apresentavam o mesmo grau neurológico de antes da cirurgia. Após sete e 90 dias de pós-operatório, 13/19 e 17/19 demonstraram melhora neurológica em comparação com o pré-operatório, respectivamente. A quantidade de contatos extradurais não influenciou na recuperação neurológica dos cães. Esses achados indicam que uma inspeção minuciosa do canal vertebral pode ser recomendada, a fim de remover o máximo de material discal extruso, evitando-se piora neurológica por compressão medular.(AU)

Aparência tomográfica dos discos intervertebrais mineralizados em cães da raça Dachshund Miniatura / Tomographic appearance of calcified intervertebral discs in miniature dachshund

Objetivou-se com este trabalho descrever os aspectos de tomografia computadorizada de discos intervertebrais de cães da raça Dachshund com um ano de idade, assim como quantificar, qualificar e localizar a mineralização nos componentes do DIV e no espaço intervertebral da coluna vertebral. Dez cães dessa raça, com um ano de idade, foram submetidos à tomografia computadorizada (TC). Nas imagens de reconstrução sagital dos animais, procedeu-se à identificação de sete DIV em cada animal, entre as vértebras T9 e L3, totalizando a análise de 70 DIV. Entre os DIV mineralizados, a presença da alteração foi determinada quanto à localização no núcleo pulposo (NP), no anel fibroso (AF) e em AF/NP. Dos 70 DIV avaliados, 45 foram considerados como mineralizados. Entre os classificados como mineralizados, 20% (9/45), 17,8% (8/45) e 62,2% (28/45) estavam localizados nas regiões do NP, AF e AF/NP, respectivamente. A mineralização encontrada neste estudo esteve caracterizada por aumento de atenuação radiográfica com densidade mineral na topografia do DIV, havendo variações da localização e da radiodensidade de área calcificada entre animais e DIV no mesmo indivíduo. As alterações relativas à mineralização dos DIV podem ser classificadas tomograficamente quanto à localização da alteração no disco em AF, NP e AF/NP.(AU)

The objective was to describe CT imaging of calcified IVD in one-year-old Dachshunds, as well as describe the location in the soft tissue structures that make up the IVD. Ten one-year-old dogs underwent computed tomography (CT). In sagittal reconstruction images of animals, the identification of 70 IVD present in the spaces between L3 - T9 were performed. Among calcified IVDs, the presence of the change was determined as the location in the nucleus pulposus (NP), annulus fibrosus (AF) and both. Of a total of 70 evaluated IVDs, 45 were found to calcified and 25 not calcified. Among calcified discs, location was determined in nine (20%) in NP, eight discs (17.77%) in AF and 28 (62, 23%) NP & AF. Calcification in this study was characterized by increased radiographic attenuation mineral density in the IVD topography. There is variation in the location and radiodensity of calcified areas between animals and IVD in the same individual. Changes related to IVD calcification can be classified as tomographic location in AF, NP and NP & AF.(AU)

Uric acid versus non-uric acid renal stones: in vivo differentiation with spectral CT.

AIM: To differentiate uric acid from non-uric acid renal stones based on their spectral attenuation values. MATERIALS AND METHODS: The present study was approved by the institutional review board and the need for informed consent was waived. Thirty-three consecutive patients (21 men, 12 women; mean age 55 years) with symptomatic urolithiasis underwent dual-energy computed tomography (DECT) using a second-generation dual-source CT system. Stone composition was assessed by means of chemical analysis after extraction or spontaneous expulsion. The composition of one stone was considered to represent all remaining stones in patients presenting with more than one stone. Image-domain virtual monoenergetic images were generated from the dual-energy datasets. One radiologist evaluated stone attenuation values from 40 to 190 keV; attenuation curves were created and 40/190 keV attenuation ratios calculated. Qualitative evaluation of the spectral attenuation curves was also performed. Imaging findings were compared with laboratory results. RESULTS: Sixty-two stones were considered in 33 patients (mean diameter 6.5 mm). Fifteen of the 62 stones were mainly composed of uric acid and 47/62 of cysteine or calcium oxalates/phosphates. Forty to 190 keV attenuation ratios were significantly lower for uric acid stones (mean 0.87±0.3) than for non-uric acid stones (mean 3.80±0.6; p<0.0001). Accuracy was 100% with a cut-off value of 1.76. Qualitative analysis of spectral attenuation curves showed unique shapes for uric acid and non-uric acid stones. CONCLUSIONS: Spectral CT quantitatively and qualitatively differentiates uric acid from non-uric acid stones.

Observações clinicocirúrgicas em fraturas e luxações vertebrais toracolombares em cães / Clinical and surgical observations in thoracolumbar spinal fractures and luxations in dogs

Objetivou-se avaliar as implicações e os resultados obtidos frente aos tratamentos conservativo e cirúrgico em 37 cães sem distinção de sexo, raça ou idade, que apresentaram fraturas e luxações vertebrais (FLV) toracolombares. Cada paciente passou por anamnese, seguida por exames clínico, neurológico e radiográficos para se obter o diagnóstico neuroanatômico e a severidade da lesão. Os cães foram submetidos ao tratamento conservativo ou cirúrgico de acordo com as características de cada caso. Dos 37 animais deste estudo, 37,7% tinham percepção de dor profunda (PDP) intacta. Vinte animais foram submetidos ao tratamento conservativo, 14 ao tratamento cirúrgico e três animais faleceram antes do tratamento. Dos animais que mantiveram a PDP intacta, a taxa de recuperação foi de 100%, com tempo de recuperação total variando de sete a 75 dias, ao passo que, dos animais que perderam a PDP, nenhum recuperou a deambulação voluntária. Dos 14 animais operados, 42,8% apresentaram alterações vertebrais não visibilizadas ao exame radiográfico convencional, sendo necessária mudança na técnica cirúrgica planejada em 14,2% desses casos. Nos animais que mantiveram a PDP, não houve diferença significante nas taxas nem no tempo de recuperação da deambulação voluntária, independentemente do tipo de tratamento. Entretanto, houve diferença altamente significante entre os graus que mantiveram a PDP intacta com os animais que perderam a percepção de dor profunda em relação à taxa de recuperação. A taxa de eutanásia foi menor que em outros trabalhos descritos na literatura. Dos animais sem PDP submetidos ao tratamento conservativo ou cirúrgico, 31,25% readquiriram a capacidade de caminhar sem recuperar a PDP, sendo essa deambulação involuntária atribuída ao caminhar espinal.

Our aim of this paper was to study the implications and results associated with conservative and surgical treatment of dogs with thoracolumbar Vertebral fractures and luxations (VFL) so that the information obtained can be used to obtain guidelines that lead to greater recovery rates. For such, 37 dogs presenting VFL were examined, without distinction of sex, breed, or age. Each patient had detailed history obtained, followed by clinical, neurologic and radiographic exams for obtaining neuroanatomic location and severity of the nervous and vertebral lesions. The dogs were submitted to conservative or surgical treatment according to each case. Deep pain perception (DPP) was intact in 37.7% of the 37 dogs in this study. Twenty dogs received conservative treatment, 14 underwent surgery and three dogs died before treatment. Of the dogs with intact DPP, 100% recovered, with a recovery time varying between seven and 75 days, while none of the dogs that lost DPP recovered voluntary motion. Of the 14 dogs submitted to surgery, 42.8% had vertebral lesions that weren't detected using conventional radiography, which demonstrates the low sensitivity of this exam, and led to a need for a change in the planned surgical technique in 14.2% of these cases. In dogs with intact DPP there was no significant difference in time or rate of recovery of voluntary motion, regardless of the treatment method. However, there was a highly significant difference between the dogs which retained and lost DPP regarding recovery rates. The percentage of euthanasia was less than in other reports, probably due to the greater awareness and protectiveness of owners nowadays. Of the dogs without DPP treated either surgically or conservatively, 31.25% recovered ambulation without recovery of DPP, characterizing this ambulation as involuntary, attributed to spinal walk.

Intraoperative ultrasonography of the vertebral canal in dogs / Ultrassonografia intraoperatória do canal vertebral em cães

Intraoperative ultrasound (IOS) can provide details on various conditions of the spinal cord and vertebral canal. The aim of the present study was to evaluate the feasibility of using IOS in dogs undergoing spinal surgery and to describe the main findings. The vertebral canal of 21 dogs was examined with intraoperative ultrasonography: 13 underwent spinal surgery for removal of herniated intervertebral disc material, three for stabilization of vertebral fracture/luxation, two for removal of vertebral neoplasia, and three for cauda equina decompression. Particular attention was given to signs of cord compression. Intraoperative ultrasonography was feasible and useful in dogs undergoing surgery for spinal cord or cauda equina decompression and fracture stabilization. It was not paramount for locating the compression when this had been done via computed tomography (CT), but it showed alterations in spinal cord parenchyma not observed on CT and also confirmed adequate decompression of the spinal cord. The main advantages of intraoperative ultrasonography were estimation of vascularization and extent of spinal cord lesion. Most importantly, it allowed real time evaluation of the spinal cord and vertebral canal, which permits the modification of the surgical procedure.

A ultrassonografia intraoperatória pode fornecer detalhes sobre diversas condições da medula espinal e canal vertebral. O objetivo deste trabalho foi avaliar a viabilidade da utilização da ultrassonografia intraoperatória em cães submetidos à cirurgia espinomedular e descrever os principais achados. O canal vertebral de 21 cães foi examinado com ultrassonografia intraoperatória - 13 foram submetidos à cirurgia para remoção de material de disco herniado, três para estabilização de fratura/luxação vertebral, dois para remoção de neoplasia vertebral e três para descompressão de cauda equina. Atenção especial foi dada a sinais de compressão medular. A ultrassonografia intraoperatória foi viável e útil em cães submetidos a cirurgia para descompressão da medula espinal ou cauda equina e estabilização de fratura. Apesar de não ser indispensável para localização da compressão quando isso havia sido feito através de tomografia computadorizada (TC), demonstrou alterações no parênquima que não haviam sido observadas pela TC e confirmou a adequada descompressão da medula espinhal. As principais vantagens da ultrassonografia intraoperatória foram: estimar a vascularização e a extensão da lesão medular. Principalmente, permitiu uma avaliação em tempo real da medula espinal e do canal vertebral, o que permite modificação do procedimento cirúrgico.

Anestesia epidural na cirurgia descompressiva lombossacral de cães / Epidural anesthesia in the lumbosacral decompressive surgery in dogs

Objetivou-se avaliar viabilidade, eficácia, vantagens e desvantagens da anestesia epidural lombossacral, junto à anestesia geral inalatória, em cirurgias de descompressão da cauda equina, tendo em vista que essas cirurgias são procedimentos longos e dolorosos. Para isso, foram utilizados 11 cães adultos com sinais clínicos de síndrome da cauda equina, que foram submetidos à anestesia geral inalatória e deixados no estágio anestésico mais superficial. Depois, realizou-se a anestesia epidural em seis dos 11 pacientes, por punção espinhal em L7-S1, com bupivacaína a 0,5 por cento. Os parâmetros fisiológicos (cardíacos, vasculares, respiratórios, temperatura corporal e glicemia) foram aferidos antes da medicação pré-anestésica, 10 minutos após esta, 30 minutos depois da epidural, depois da laminectomia, assim como após 60 minutos e 90 minutos da epidural, tanto no grupo com epidural como naquele sem esta (controle). Os animais que possuíam bloqueio epidural apresentaram redução significativa no consumo de anestésico inalatório e no tempo de extubação, não apresentando déficits neurológicos causados pela anestesia epidural, quando comparados com o grupo-controle. Conclui-se que a técnica de anestesia epidural é eficiente e vantajosa na realização de cirurgias descompressivas lombossacrais, pois proporciona menor risco anestésico para o animal...

The objective of this study was to evaluate the feasibility, effectiveness, advantages and disadvantages of lumbosacral epidural anesthesia with inhalational anesthesia in surgical decompression of the cauda equina, since this surgery has a lengthy and painful recovery from anesthesia. For this, 11 adult dogs which showed clinical signs of cauda equina syndrome were used. After the epidural, anesthesia in six of 11 patients, spinal tap in L7-S1, with bupivacaine 0.5 percent anesthetic was used. Physiological parameters (heart, vascular, respiratory and body temperature) were measured before, 10 minutes after, and 30 minutes after the epidural, after the laminectomia and 60 minutes and 90 minutes after the epidural, both in the epidural group and the control group. Animals that had ep showed a significant reduction in consumption of inhaled anesthetic and time of extubation. There were no neurological deficits caused by epidural anesthesia compared with the group without epidural anesthesia. It was concluded that the epidural anesthesia technique is effective in lumbosacral decompression surgeries, leading to a lower risk for the anesthetized animal...

Cardiovascular and respiratory alterations in dogs during cervical myelography using ioversol / Alterações cardiovasculares e respiratórias durante a mielografia cervical com ioversol em cães

Subarachnoid infusion of most contrast mediums and the steps involved in performing a cervical myelography have adverse affects that can discourage its use in the radiographic diagnosis of spinal cord diseases. Thus, the cardiovascular and respiratory alterations associated with neck flexion, subarachnoid puncture, and cerebrospinal fluid drainage during subarachnoid infusion of ioversol (320mgI/mL) in dogs under general anesthesia using isoflurane were evaluated. The dogs received subarachnoid infusion of autologous cerebrospinal fluid kept at 38°C - control group (GC); ioversol 0.3mL/kg at 25°C (GI25) and ioversol 0.3mL/kg heated to 38°C (GI38). Each dog had its heart rate (HR), systolic arterial pressure (SAP), diastolic arterial pressure (DAP), respiratory rate (RR), oxyhemoglobin saturation (SaO2) and electrocardiography readings (PR and QT intervals) recorded. Group comparisons showed no statistical difference regarding neck positioning, subarachnoid puncture, and subarachnoid infusion of contrast medium on HR, RR and SaO2, cardiac rhythm or conduction. However, isoflurane significantly increased PR and QT intervals. Based on these findings, it is concluded that the steps involved in cervical myelography and the use of ioversol 320mgI/mL at 0.3mL/kg (25ºC and 38ºC) during cervical myelography did not result in relevant cardiovascular and respiratory alterations, except for an elevation in arterial pressure after injection of ioversol.

Os efeitos adversos provocados pela infusão subaracnoidea de meios de contraste e da mielografia cervical podem desencorajar sua aplicação como alternativa para o diagnóstico radiológico de doenças da medula espinhal. Assim, avaliaram-se os efeitos cardiovascular e respiratório dos procedimentos técnicos necessários à mielografia cervical e da infusão subaracnoidea do ioversol - 320mgI/mL - em cães anestesiados com isofluorano. Foi realizada infusão do líquido cefalorraquidiano autólogo mantido a 38°C - grupo-controle (GC); infusão do ioversol 0,3mL/kg a 25°C (GI25); e infusão do ioversol 0,3mL/kg aquecido a 38°C (GI38). Estudaram-se: pressões arteriais sistólica (PAS) e diastólica (PAD), frequências cardíaca (FC) e respiratória (FR), saturação de oxi-hemoglobina (SaO2) e medidas eletrocardiográficas (intervalos PR e QT). Não houve influência significativa do posicionamento, da punção ou da infusão subaracnoidea do ioversol sobre FC, FR, SaO2, ritmo e condução cardíaca, entretanto o isofluorano aumentou significativamente os intervalos PR e QT. Concluiu-se que a mielografia cervical com ioversol 320mgI/mL, nas temperaturas de 25ºC e 38ºC e na dose de 0,3mL/kg, não causa arritmias cardíacas nem alterações cardiovasculares e respiratórias relevantes, exceto elevação da pressão arterial após a injeção do contraste.

Lesão iatrogênica meningomedular em um cão submetido à espondilectomia ventral para tratamento de extrusão crônica de disco intervertebral cervical / Iatrogenic meningo-medullary lesion in a dog undergoing ventral spondylectomy for treatment of a chronic cervical intervertebral disc extrusion

Objetivou-se descrever a ocorrência de lesão iatrogênica meningomedular em um cão da raça Dachshund, macho, oito anos de idade, apresentando tetraparesia flácida há dois meses, que foi submetido à espondilectomia ventral para tratamento de extrusão crônica de disco intervertebral cervical. Durante remoção do material de disco herniado com removedores de tártaro e pinça hemostática, observou-se severa hemorragia dos seios venosos, descolamento meníngeo com laceração medular e imediata queda dos parâmetros cardiovasculorrespiratório, com evolução para a morte ante a ausência de respostas às medidas de ressuscitação cardiopulmonar.

Our objective was to describe the occurrence of an iatrogenic meningo-medullary lesion and the trans-surgical complications in an eight-year-old male Daschund, which presented flacid tetraparesis for two months, and underwent ventral spondylectomy to treat a chronic cervical intervertebral disc extrusion. During the removal of the herniated disc with dental picks and hemostatic forceps, we observed meningeal dislocation with medullary laceration and an immediate reduction in cardiovasculatory and respiratory parameters, which progressed to a cardiorespiratory arrest that was non-responsive to medication and finally led to the death of the patient.

Substituição da agulha isolada para eletroneuroestimulação pela agulha metálica de cateter intravenoso, na verificação da punção epidural, em cães / Replacement of an insulated electric neurostimulation spinal needle with an intravenous catheter metal needle to confirm correct epidural puncture in dogs

Utilizou-se o eletroneuroestimulador como um meio adequado para confirmação do posicionamento da ponta de agulhas isoladas no espaço epidural lombossal, por meio de pequenas estimulações elétricas que provocam contrações dos músculos inervados, empregando-se agulha de cateter intravenoso em vez de agulha isolada para a eletroneuroestimulação. Foram utilizados 40 cães para procedimentos cirúrgicos ortopédicos, abdominais ou retroumbilicais que necessitassem de bloqueio epidural. Durante a introdução gradativa da agulha no canal vertebral lombossacral, estímulos elétricos de 1,5 a 0,5mA foram aplicados. Diante de contrações musculares em membros pélvicos, cauda e/ou ânus, as quais foram consideradas respostas condizentes com o correto posicionamento da agulha no espaço epidural, estimulando raízes da cauda equina, procedeu-se à administração da associação de lidocaína, bupivacaína e tramadol. Constatou-se a adequada anestesia epidural mediante relaxamento do esfíncter anal, ausência dos reflexos patelares e flexor e a analgesia na área cirúrgica abdominal. Conclui-se que o método foi eficaz para a confirmação do correto posicionamento da agulha no espaço epidural durante a realização dessa anestesia regional em cães.

The electric neurostimulator was used as an adequate mean for the confirmation of the correct placement of the tip of the insulated needles in the lumbosacral epidural space through small electric stimulations causing muscle contractions by using an intravenous catheter needle instead of an electric neurostimulation insulated needle. Forty dogs were selected based on the need for an epidural block for orthopedic, abdominal or retroumbilical surgical procedures. Electrical stimuli were applied from 1.5 to 0.5mA during a gradual introduction of the needle into the lumbosacral vertebral canal. Responses that were considered as indicative of the correct placement of the needle and stimulation of the cauda equina were muscle contractions in the pelvic limbs, tail or anus. An association of lidocaine, bupivacaine and tramadol was then administered. Adequate epidural anesthesia was confirmed through relaxation of the anal sphincter, absence of patellar and flexor reflexes and analgesia of the abdominal surgical area. The authors were able to conclude that the method used was effective to confirm the correct needle placement in the epidural space during regional anesthesia in dogs.

Type-3 metabotropic glutamate receptors regulate chemoresistance in glioma stem cells, and their levels are inversely related to survival in patients with malignant gliomas.

Drug treatment of malignant gliomas is limited by the intrinsic resistance of glioma stem cells (GSCs) to chemotherapy. GSCs isolated from human glioblastoma multiforme (GBM) expressed metabotropic glutamate receptors (mGlu3 receptors). The DNA-alkylating agent, temozolomide, killed GSCs only if mGlu3 receptors were knocked down or pharmacologically inhibited. In contrast, mGlu3 receptor blockade did not affect the action of paclitaxel, etoposide, cis-platinum, and irinotecan. mGlu3 receptor blockade enabled temozolomide toxicity by inhibiting a phosphatidylinositol-3-kinase/nuclear factor-κB pathway that supports the expression of O(6)-methylguanine-DNA methyltransferase (MGMT), an enzyme that confers resistance against DNA-alkylating agents. In mice implanted with GSCs into the brain, temozolomide combined with mGlu3 receptor blockade substantially reduced tumor growth. Finally, 87 patients with GBM undergoing surgery followed by adjuvant chemotherapy with temozolomide survived for longer time if tumor cells expressed low levels of mGlu3 receptors. In addition, the methylation state of the MGMT gene promoter in tumor extracts influenced survival only in those patients with low expression of mGlu3 receptors in the tumor. These data encourage the use of mGlu3 receptor antagonists as add-on drugs in the treatment of GBM, and suggest that the transcript of mGlu3 receptors should be measured in tumor specimens for a correct prediction of patients' survival in response to temozolomide treatment.

Abatacept (CTLA-4IG) treatment reduces the migratory capacity of monocytes in patients with rheumatoid arthritis.

OBJECTIVE: The binding of abatacept (CTLA-4Ig) to the B7 ligands CD80 and CD86 prevents the engagement of CD28 on T cells and thereby prevents effector T cell activation. In addition, a direct effect of CTLA-4Ig on antigen-presenting cells (APCs) could contribute to the therapeutic effect. To further elucidate the mechanism of CTLA-4Ig, we performed phenotype and functional analyses of APCs in patients with rheumatoid arthritis (RA) before and after the initiation of CTLA-4Ig therapy. METHODS: Peripheral blood mononuclear cells were analyzed before and at 2 and 4 weeks after the initiation of CTLA-4Ig therapy. Proportions of APCs were determined by flow cytometry. CD14+ monocytes were further analyzed for the expression of costimulatory and adhesion molecules and for their transendothelial migratory capacity in vitro. In addition, CD14+ monocytes from healthy controls were analyzed for their migratory and spreading capacity. RESULTS: Proportions and absolute numbers of monocytes were significantly increased in RA patients treated with CTLA-4Ig. The expression of several adhesion molecules was significantly diminished. In addition, monocytes displayed a significant reduction in their endothelial adhesion and transendothelial migratory capacity upon treatment with CTLA-4Ig. Likewise, isolated monocytes from healthy controls revealed a significant reduction in their migratory and spreading activity after preincubation with CTLA-4Ig or anti-CD80 and anti-CD86 antibodies. CONCLUSION: We describe direct effects of CTLA-4Ig therapy on phenotype and functional characteristics of monocytes in RA patients that might interfere with the migration of monocytes to the synovial tissue. This additional mechanism of CTLA-4Ig might contribute to the beneficial effects of CTLA-4Ig treatment in RA patients.

Heme oxygenase-1 end-products carbon monoxide and biliverdin ameliorate murine collagen induced arthritis.

OBJECTIVES: Heme oxygenase-1 (HO-1) which degrades Heme to free iron, biliverdin and carbon monoxide (CO) plays an important role in inflammation. There are, however, conflicting data concerning the role of HO-1 in rheumatoid arthritis (RA) and the therapeutic potential of individual heme degradation products remains to be determined. We therefore investigated the effect of CO and biliverdin upon therapeutic administration in the murine collagen induced arthritis (CIA) model of RA. METHODS: CIA was induced in DBA/1 mice. Anti-CII antibody levels were determined by ELISA. Mice were scored for paw swelling and grip strength. After the first clinical signs of arthritis one group of animals was treated with biliverdin, the second group was treated with CO. After 60 days all animals were sacrificed and analysed for histomorphological signs of arthritis. RESULTS: All animals immunised with CII developed serum anti-CII antibodies. Antibody levels were decreased in the CO-treated group. Both, Biliverdin and the CO-treated animals, showed an improvement in clinical disease activity. Histological analysis revealed significantly less inflammation, erosion and reduced numbers of osteoclasts in CO-treated animals only, whereas cartilage degradation was prevented in both biliverdin and CO-treated animals. CONCLUSIONS: Our data demonstrate a beneficial effect of CO, in particular, and biliverdin, on inflammation and bone destruction in the CIA mouse model.

The origin recognition complex subunit, ORC3, is developmentally regulated and supports the expression of biochemical markers of neuronal maturation in cultured cerebellar granule cells.

The origin recognition complex (ORC) regulates DNA replication. However, some members of the ORC core, such as ORC3 and ORC5, have been implicated in neuronal maturation. In cultured cerebellar granule cells (CGCs), ORC3 mRNA and protein levels increased from 6 to 8days in vitro, a time that coincided with the maximal development of the dendritic arbor. In contrast, expression of ORC5 remained low throughout CGC maturation. Activation of type-4 metabotropic glutamate receptors with the selective enhancer, PHCCC, during a critical time-window (from 4 to 6days in vitro) anticipated the developmental peak of ORC3, increased the expression of two proteins associated with neuronal maturation, i.e. the mitogen-associated protein-2 (MAP-2) and postsynaptic density-95 (PSD-95), as well as dendritic length. siRNA-induced ORC3 knockdown reduced MAP-2 and PSD-95 expression on its own and abrogated the action of PHCCC. We examined whether the maturational effects of ORC3 were mediated by changes in the activity of the monomeric GTP-binding protein, Rho, which is known to regulate granule cell morphology. ORC3 knockdown increased the levels of the GTP-bound active form of Rho, whereas exposure to PHCCC reduced Rho activation. The action of PHCCC was largely attenuated in cultures deprived of ORC3. Finally, granule cell exposure to the Rho-associated kinase inhibitor, Y-27632, abolished the lowering effect of ORC3 knockdown on MAP-2 expression, and increased dendritic length. These data suggest that ORC3 supports neuronal maturation by inhibiting the Rho signaling pathway, and mediates the differentiating activity of mGlu4 receptors in cultured cerebellar granule cells.

Treg and lupus.

Autoimmune diseases such as systemic lupus erythemathodes (SLE) display quantitative and/or qualitative deficiencies of CD4+CD25+ regulatory T cells (Treg). This might contribute to the immune dysregulation and break-down of peripheral tolerance mechanisms. In addition novel subsets of Treg might arise under specific autoimmune conditions as an effort to counteract autoimmunity. Advances in our understanding of how to characterise and manipulate Treg in patients with autoimmune diseases are required in order to develop new therapeutic strategies.