Perivascular Cuffing Research Articles

Rosuvastatin revert memory impairment and anxiogenic-like effect in mice infected with the chronic ME-49 strain of Toxoplasma gondii.

The aim of this study was to investigate the effect of rosuvastatin treatment on memory impairment, and anxiogenic-like effects in mice chronically infected with Toxoplasma gondii. For this, Balb/c mice were infected orally with chronic ME-49 strain of Toxoplasma gondii. Oral treatment with rosuvastatin (40mg/kg/day) started on the 51st day post-infection and was performed daily for 21 days. After completion of treatment, anxiety-like effects and locomotion were investigated in the open field (OF) test, whereas novel object recognition (NOR) test was used for evaluation of short- and long-term memory. At the end of the experiments, the brain was collected for Toxoplasma gondii DNA quantification and histopathological analysis. Infection with ME-49 strain decreased the time spent in the center of OF, indicating an anxiogenic effect, without affecting total and peripheral locomotion. Rosuvastatin treatment inhibited the change in the center time. Besides, pharmacological treatment increased total and central locomotion in both non-infected and infected animals. Infection also impaired both short- and long-term memory in the NOR test, and these effects were reverted by rosuvastatin treatment. In addition to effects in behavioral changes, rosuvastatin also reduced parasite load in the brain and attenuated signs of brain inflammation such as perivascular cuffs, inflammatory cell infiltration and tissue damage. These findings indicate for the first time the efficacy of rosuvastatin in treatment of memory impairment and anxiogenic effect evoked by infection with Toxoplasma gondii. These effects might be mediated by reduced cyst load, which in turn decrease inflammation and damage in the brain.

Recovery of Synthetic Zika Virus Based on Rio-U1 Isolate Using a Genetically Stable Two Plasmid System and cDNA Amplification

In 2016, the world experienced the unprecedented Zika epidemic. The ZIKV emerged as a major human pathogen due to its association with the impairment of perinatal development and Guillain–Barré syndrome. The occurrence of these severe cases of Zika points to the significance of studies for understanding the molecular determinants of flavivirus pathogenesis. Reverse genetics is a powerful method for studying the replication and determinants of pathogenesis, virulence, and viral attenuation of flaviviruses, facilitating the design of vaccines and therapeutics. However, the main hurdle in the development of infectious clones is the instability of full-length cDNA in Escherichia coli. Here, we described the development of a genetically stable and efficient infectious clone based on the ZIKV Rio-U1 isolated in the 2016 epidemic in Brazil. The employed strategy consisted of cloning the viral cDNA genome into two stable plasmid subclones and obtaining a high-quality cDNA template with increment in DNA mass for in vitro transcription by PCR amplification. The strategy for developing a ZIKV infectious cDNA clone designed in this study was successful, yielding a replicative and efficient clone-derived virus with high similarities with its parental virus, Rio-U1, by comparison of the proliferation capacity in mammal and insect cells. The infection of AG129 immunocompromised mice caused identical mortality rates, with similar disease progression and morbidity in the animals infected with the parental and the cDNA-derived virus. Histopathological analyses of mouse brains infected with the parental and the cDNA-derived viruses revealed a similar pathogenesis degree. We observed meningoencephalitis, cellular pyknosis, and neutrophilic invasion adjacent to the choroid plexus and perivascular cuffs with the presence of neutrophils. The developed infectious clone will be a tool for genetic and functional studies in vitro and in vivo to understand viral infection and pathogenesis better.

Evaluation of MMP-9 and iNOS expressions in sheep with encephalitic listeriosis

This study aimed to correlate Matrix Metalloproteinase-9 and iNOS expressions with the severity of histopathological findings in tissue samples taken from sheep with encephalitic listeriosis. Thus, the role of these molecules in the pathogenesis of the disease can be elucidated. After systemic necropsy, tissue samples of adult sheeps with meningoencephalitis were investigated by the culture, histopathological and immunohistochemical methods in the presence of Listeria spp. isolation from tissues was performed in accordance with the USDA-FSIS method with some modifications. Tissue samples were fixed in a 10% buffered formaldehyde solution. Following routine procedures, tissue sections at 5 μm were stained with Hematoxylin and Eosin, investigated under light microscope and photographed. Immunohistochemical staining was performed on the tissues using the avidin-biotin immune peroxidase complex method. Listeria spp. were obtained in 20 (83.3%) of 24 tissue samples with the presence of bright grey-black centred smooth colonies on Listeria Selective Agar and identified as Listeria monocytogenes through the phenotypically supportive tests. Liquefaction necrosis, purulent meningoencephalitis, perivascular cuffing, microabscesses and glial nodules were the most important histopathological findings. MMP-9 immunpositive reactions were observed in the cytoplasm of microglial cells and neurons in areas where inflammatory and necrotic areas are concentrated in medulla oblongata and pons. In perivascular cuffing areas, immune reactions in endothelial cells were detected. We detected iNOS positive reactions in the medulla oblangata and pons, especially in inflammatory cells in the microabscesses. Consequently, a positive correlation (p < 0.05) was found between MMP-9 expression and the severity of histopathological findings in sheep with encephalitic listeriosis. expression. In addition, we found that iNOS expression increased in parallel with the increase in MMP-9

COVID-19 vasculitis and novel vasculitis mimics.

COVID-19 has been occasionally linked to histologically confirmed cutaneous vasculitis and a Kawasaki-like vasculitis, with these entities generally having minimal or no lung involvement and a good prognosis. Unlike these vasculitis types, patients with severe COVID-19 pneumonia can develop cutaneous vasculitis-like lesions and systemic arterial and venous thromboemboli, including cryptogenic strokes and other vasculopathy features. Proposed underlying mechanisms for these severe manifestations have encompassed immune dysregulation, including an anti-phospholipid syndrome-like state, complement activation, viral dissemination with direct systemic endothelial infection, viral RNAaemia with immunothrombosis, clotting pathway activation mediated by hypoxaemia, and immobility. In this Viewpoint, we highlight how imaging and post-mortem findings from patients with COVID-19 indicate a novel thrombosis in the pulmonary venous territory distal to the alveolar capillary bed, a territory that normally acts as a clot filtration system, which might represent an unappreciated nidus for systemic microembolism. Additionally, we suggest that this mechanism represents a novel vasculitis mimic related to COVID-19 that might lead to cryptogenic strokes across multivessel territories, acute kidney injury with haematuria, a skin vasculitis mimic, intestinal ischaemia, and other organ ischaemic manifestations. This finding is supported by pathological reports of extensive pulmonary venular thrombosis and peripheral organ thrombosis with pauci-immune cellular infiltrates. Therefore, severe COVID-19 pneumonia with extensive pulmonary intravascular coagulopathy might help to explain the numerous systemic complications of COVID-19, in which the demonstration of direct organ infection has not adequately explained the pathology.

Encephalitozoonosis in Rabbit (Oryctolagus cuniculus)

Background: Encephalitozoonosis is caused by the protozoan Encephalitozoon cuniculi, in rabbits, and can affect humans. The disease can be fatal and difficult to diagnose. It can be asymptomatic or cause vestibular neurological disease, paralysis, uveitis in addition to chronic kidney disease in rabbits. The transmission of the microorganism's spores occurs by ingestion, inhalation, or by the transplacental route. The aim of this work is to report a case of encephalitozoonosis in a pet rabbit (Oryctolagus cuniculus).Case: An Oryctolagus cuniculus with a history of paraparesis of the thoracic and pelvic limbs was referred for necropsy, the evolution of the clinical picture happened in one day. After death, a necropsy was performed. Organ fragments were collected, fixed, and processed routinely for histology. Macroscopically, there was evidence of hepatic lobes, without injury to the other organs. Microscopically it was observed in the white and gray substance of the telencephalon multiple circumscribed granulomas composed of a necrotic center surrounded by macrophages, giant multinucleated cells in addition to lymphocytes and plasmocytes in the periphery, delimited by fibrous connective tissue. Around the vessels, perivascular cuffs with two to four layers of lymphocytic infiltrate were observed. Besides, special staining of Schiff's Periodic Acid (PAS) and Ziehl-Neelsen was performed, in which numerous cylindrical, eosinophilic structures of approximately 2.5 x 1.0 µm were observed, compatible with E. cuniculi spores. Besides, histiocytic lymphoblasts pericoronitis was noted in the liver. There were no relevant changes in the kidney.Discussion: The diagnosis of encephalitozoonosis in rabbits was based on clinical and anatomopathological findings. Tetraparesis was the predominant sign in the present case and was justified by telencephalic lesions. This clinical sign is included in the literature but is less common than the syndrome such as head tilt and paralysis. The diagnosis of the disease is usually made by post-mortem examination when it is possible to identify the spores in the lesions. Multifocal granulomatous encephalitis was the most significant finding in this case, which is also consistent with other studies. The pathogenesis of granulomatous lesions is still controversial. It is known that spores allow phagocytosis by macrophages, which induce the production of interleukins and other cytokines by TCD4 + lymphocytes, thereby activating the action of TCD8 + (cytotoxic) lymphocytes. Natural killer cells, granulocytes, other macrophages, and B lymphocytes are also recruited. Although there is such an inflammatory response, the antibodies produced are not efficient to eliminate the agent from the host organism, however, they contribute to the process of opsonization and consequent phagocytosis, facilitating the destruction of the microsporidium by macrophages. The neurological form was predominant in this case, with no chronic or ocular renal forms, possibly due to the rapid clinical evolution. Special stains were useful for visualizing intralesional spores. Although PAS staining is considered to be of little use, it was relevant in this case. The visualization of the agent made it possible to distinguish differential diagnoses, among them vestibular syndrome secondary to otitis due to pasteurellosis, toxoplasmosis, neoplasms, traumas, or diseases of the spine. Thus, a diagnosis of encephalitozoonosis was made in a rabbit through clinical and anatomopathological correlation using Ziehl-Neelsen and PAS stains.Keywords: granuloma, Encephalitozoon cuniculi, central nervous system.Descritores: granuloma, Encephalitozoon cuniculi, sistema nervoso central.Título: Encefalitozoonose em coelho (Oryctolagus cuniculus)

Encephalic Toxoplasm in a White-Eared Possum (Didelphis albiventris)

Background: Toxoplasmosis is caused by Toxoplasma gondii, an obligate intracellular protozoan that belongs to the Aplicomplexa phylum, coccidian subclass, and affects all warm-blooded animals. The role of opossums in the epidemiology of toxoplasmosis in Brazil is not fully understood, and there are very few descriptions of toxoplasmosis lesions in these animals. This report describes the anatomopathological, molecular and immunohistochemical findings of a case of encephalic toxoplasmosis in free-living white-eared possum (Didelphis albiventris).Case: A young male opossum (D. albiventris), was treated at the Veterinary Hospital of Wild Animals of the University of Brasília, Federal District. The animal was apathetic, uncoordinated, reluctant to move, and had an exposed proximal fracture in the left radius and ulna with laceration of muscles and adjacent tendinous structures. Amputation on the left thoracic limb was performed followed by analgesia and antibiotic therapy. The environment is frequented by other wild animals, and stray cats have access to the patio of the building. Twenty-five days after arriving at the hospital, the animal was found dead in its cage. After death, a necropsy was performed. Organ fragments from the abdominal cavity, thoracic and central nervous system were collected, processed routinely for histology and stained with hematoxylin and eosin. Macroscopic lesions in the central nervous system were not observed. On microscopy, the brain showed moderate random glial nodules throughout the neuropil associated with the presence of spherical to elongated parasitic cysts of about 20 µm, with a thin wall and with its interior full of bradyzoites, consistent with Toxoplasma gondii. There was also moderate fibrinoid necrosis and moderate multifocal lymphoplasmacytic infiltrate surrounding the blood vessels (perivascular cuffs) To investigate the etiology of the brain injury, brain sections were subjected to immunohistochemistry (IHC) and real-time polymerase chain reaction (qPCR) technique for detection of T. gondii and Neospora caninum. Immunostaining for T. gondii in the cyst wall and in bradyzoites and negative immunostaining for N. caninum. qPCR was positive for T. gondii and negative for N. caninum.Discussion: Diagnosis of encephalic toxoplasmosis in a Didelphis albiventris was possible based on histopathological, immunohistochemical and molecular findings. The morphological classification of the brain lesion was important for the diagnosis. Brain toxoplasmosis in opossums usually results in focal areas of malacia on macroscopy and focally extensive necrosis on microscopy, neutrophil infiltrate, calcified necrotic material, and perivascular cuffs of lymphocytes and plasma cells. In the present case, similar histopathological lesions were noted, but no significant macroscopic changes were observed. The etiology here was defined by immunohistochemistry and qPCR, techniques proven to be useful and with good specificity for diagnosing toxoplasmosis in mammals. It is believed that the positive immunohistochemical and molecular result for Toxoplasma gondii together with the negative result for Neospora caninum were conclusive for the diagnosis. Thus, we demonstrate here a post mortem diagnosis of toxoplasmosis in a free-living synanthropic opossum and the use of anatomopathology, immunohistochemistry and real-time polymerase chain reaction as a diagnostic option for this disease in opossums. Keywords: Toxoplasma gondii, marsupial, immunohistochemistry, Real time PCR, protozoal encephalitis.Título: Toxoplasmose encefálica em um gambá-de-orelha-branca (Didelphis albiventris)Descritores: Toxoplasma gondii, marsupial, imuno-histoquímica, Real time PCR, encefalite protozoal.

Transplacental transmission of Neospora caninum to lambs in successive pregnancies of naturally infected sheep in Southern Brazil.

This study aims to report the detection of N. caninum DNA in a newborn lamb (1) with neurological signs and congenital neosporosis and in a stillborn lamb (2), both born from the same ewe in a herd of Southern Brazil. The lambs were born during different pregnancies of a Suffolk ewe seropositive to N. caninum and seronegative to T. gondii. Histopathological lesions were observed only in the central nervous system of the lambs. The newborn lamb (1) showed mild and focal gliosis in the frontal lobe. In the hippocampal region of the stillborn lamb (2), lymphoplasmacytic perivascular cuffs and N. caninum cysts were observed in the cytoplasm of neurons and confirmed by IHC. PCR was performed using brain samples to detect the protozoa N. caninum and Toxoplasma gondii. The infection with N. caninum was confirmed in the newborn lamb (1) by PCR and in the stillborn lamb (2) by histopathology, immunohistochemistry, and PCR tests.

Evaluation of correlation of EIV(H3N8) replication and pathological changes in lungs of BALB/c mice

Equine influenza (EI) is a highly contagious viral respiratory disease caused by the equine influenza virus (EIV). Frequent antigenic changes of EIV warrants thorough pathological evaluation and in-vivo replication of the circulating viruses in an experimental animal model for selection of vaccine strain. In the present study, EIV (H3N8) was inoculated into BALB/c mice and subsequently, gross, histopathological and immunohistochemical examination of the lungs was performed to evaluate the correlation of virus replication with corresponding pathological changes. Microscopically, at 1dpi, lungs from mice infected with EIV revealed degeneration of epithelial tissues, whereas indirect immunoperoxidase staining (IIPT) results showed high EIV antigen positivity in the bronchial and bronchiolar epithelium and mild positivity in the infiltrating cells. Subsequently from 1 dpi to 3 dpi, the virus replicated and infected other parenchymal cells, which led to recruitment of inflammatory cells. Microscopically moderate peribronchial, peribronchiolar and perivascular cuffing of neutrophils and macrophages were observed, whereas IIPT exhibited, scanty and moderate EIV antigen positive interstitial infiltrating macrophages. At 5 and 7 dpi, severe, diffuse interstitial thickening, bronchial, bronchiolar and alveolar epithelial necrosis and denudation, perivascular and peribronchial mononuclear cells cuffing were observed, which could be correlated with IIPT as high EIV antigen positivity of infiltrating macrophages and type II pneumocytes. However, from 11 dpi onwards, microscopically, moderate interstitium thickening and perivascular and peribronchial cuffing were observed whereas, tissues were negative for EIV antigen in IIPT. Mild, focal inflammation could be observed at 14 dpi. Present findings signify the correlation of virus replication with corresponding histopathological changes, which will be useful for assessing the efficacy of anti-influenza drug molecules and EIV vaccines.

Gambaran Mikroskopis Berbagai Organ Merpati yang Menunjukkan Gejala Tortikolis dan Identifikasi Virus Penyakit Tetelo Berdasarkan Uji Agar Gel Presipitasi

This study aimed to determine the microscopic conditions of organs of pigeons that suffered from torticollis and identify the cause of disease in torticollis pigeons. Three pigeons showed torticollis symptoms were obtained as sample and marked as pigeon A, B and C, respectively. Isolation of pathogen with inoculation into embryonated pigeon eggs obtained from parents with no history of vaccination and not indicated ND. Identification of ND with agar gel precipitation (AGP) test. Observation of microscopical changes with histopathologic preparation using hematoxylin and eosin staining. Histopathological examination showed that pigeons was done severe neuritis vagus, trakheitis, pneumonia, air sacculitis, hepatitis, pankreatitis, nefritis, jejunoileitis, ileocolitis and orchitis. Perivascular cuffing found in brain. Degenerative changes found in the hepar and ren. Cardiac severe necrotic lesion, and depletion in white pulp area of spleen. Proventricular tissue showed flattening of mucosal epithelium, congestion lesions found in pulmonary tissue. The results of slow hemagglutination test of pigeon egg allantoic fluid, which tested for hemagglutination (HA) showed positive HA result with titers varying between 2, 32, and 64. Serological identification carried out with the AGP test on all culture samples against ND antiserum showing positive results of ND virus. Based on these dara, it can be concluded that the pigeons with symptoms of torticollis is caused by ND virus.

INQUISITION OF CARDIOPULMONARY AND KIDNEY LESIONS PRODUCED BY NATURAL PERACUTE AND ACUTE CLOSTRIDIUM TYPE D ENTEROTOXEMIA IN CHINKARA DEER

This study describes the clinical signs, postmortem lesions and histopathological findings in chinkara deer died during an outbreak of enterotoxaemia. The study included 7 chinkara deer. During the outbreak, 5 chinkara deer died without clinical signs while 2 chinkara deer showed clinical signs before death. Clinical signs including greenish watery diarrhea, chocolate colored urine, herding in a corner with head down, fever, anorexia and opisthotonos. After death of these 7 deer, necropsy performed and observed soft and swollen kidneys, straw color fluid in abdominal cavity, edematous and congested lungs, hydrothorax, hydropericardium, hemorrhages and ballooning of small and large intestine and watery contents in the small intestine. In per-acute cases multifocal petechial hemorrhages on jejunal mucosa and myocardium were frequently observed. The histologic changes were perivascular cuffing in lungs and proteinaceous fluid in alveoli, pulmonary edema and hemorrhagic enteritis. Disruption of renal tubular epithelial cells, necrosis and congestion observed in histopathological section of kidneys. Affected deer was treated with different drugs and supplements but unfortunately prognosis was death. Enterotoxaemia is more fatal in young deer than adult. On the basis of this study we found that enterotoxaemia clinical signs, postmortem lesions and histopathological changes in deer are same as other animals such as sheep and goats.

Early Failures of Total Knee Patients with Nickel Allergies Secondary to Carbon Fiber Debris

Aim: Currently, there are not many studies on the long-term outcomes of the specific implants used in patients with metal hypersensitivities. The purpose of this study is to report a significant number of TKA revision failures with the CFR-PEEK prosthetic knee implant with an anatomic hinge. Background: Patients can experience multiple issues following a primary Total Knee Arthroplasty (TKA). A growing body of research is finding a correlation between primary TKA failure and metal hypersensitivity, most commonly with nickel and cobalt. Because of this, knee prosthetics are being made from hypoallergenic metals, such as zirconium nitride (ZrN), to minimize the number of failures due to metal allergy. Given the relatively new development of the hypoallergenic prostheses, there is sparse data about their overall success. Objective: The purpose of this study is to report a significant number of TKA revision failures with the CFR-PEEK prosthetic knee implant with an anatomic hinge. Methods: In this study, we analyzed the sequelae of 84 patients between May 2015 to June 2020 who received a total knee revision due to a failed primary TKA with metal hypersensitivity. These patients were revised with the rotating anatomic hinge knee system, which consisted of Carbon Fiber Reinforced Poly-Ether-Ether-Ketone (CFR-PEEK implant) coated with a ZrN, Zirconium, and Chrome Nitride multi-layer surface coating with 30% polyacrylonitrile fiber content. The patients requiring an additional revision operation presented with increasing pain that showed evidence of osteolysis on plain radiographs, CT imaging, positive indium labeled WBC scans and increased uptake on bone scan. Intraoperatively, an open incisional biopsy of the surgical bed was obtained, followed by radical debridement and implantation of porex knee prosthesis. In addition, intraoperative findings included black debris that grossly stained the surrounding synovial tissue black, indicative of wear and debris from carbon fiber bearings and the anatomic hinge. Histology of the intraoperative biopsy showed the accumulation of black debris in the histiocyte, fibrosis, and perivascular cuffing of lymphocytes and multinucleated giant cells stained with black debris. Results: Of the 84 patients, 22 failed, requiring an additional revision procedure, equating to a 26 percent failure rate of the CFR-PEEK implant, averaging 25 months to failure. Conclusion: This study was meant to report the extremely high failure rate of the anatomic hinge secondary to carbon fiber debris. Perhaps carbon fiber-laden implants are not as good an option for total knee revision prosthesis because of the high failure rate compared to those with polyethylene in their hinge mechanism.

NIMG-62. AMPHETAMINE-ASSOCIATED CNS VASCULITIS MASQUERADING AS TUMOUR

Abstract Amphetamine-associated CNS vasculitis is controversial. There are few reports but none as a tumor mimic to our knowledge. We describe a pathology-proven, amphetamine-associated CNS vasculitis resembling a tumor. A 41-year-old man presented with six weeks of progressive confusion and left-sided weakness. CT head reported a right thalamic mass with vasogenic edema. Laboratory tests were unremarkable except for cannabinoids and amphetamine in urine. MRI brain showed enhancement in right frontotemporal lobe, basal ganglia, and thalamus concerning for glioblastoma multiforme. After high-dose IV dexamethasone, an initial biopsy showed reactive gliosis, perivascular lymphocytic cuffing by CD3+, rare CD20+ cells and no infection. He continued to decline. MRI 22 days after admission showed increased T2/FLAIR hyperintensity, multifocal areas of enhancement, microhemorrhages, and ischemia. High-grade glioma, lymphoma and infectious encephalitis remained on differentials. Treatment included broad-spectrum antibiotics and acyclovir. Biopsy on 23rd day showed reactive gliosis with parenchymal macrophage infiltration and perivascular cuffing with mixed inflammatory infiltrates. CSF: slightly elevated WBCs (8/µL), elevated protein (139mg/dL), normal glucose, and no infection. MRI whole spine and CT body were unremarkable. The patient was transferred to MDACC 5.5 weeks after initial presentation. He was alert with expressive aphasia, right gaze preference, left hemiplegia, and right hemiparesis. He underwent a right anterior temporal lobectomy. Pathology showed extensive cortical laminar and white matter necrosis with macrophage infiltration and microglial activation. In areas of the preserved cortex, vasculocentric lymphocytic aggregates were present and reticulin staining showed lymphocytic infiltration of the vascular walls. Immunophenotyping (CD3, CD20) showed an almost pure T-cell population. The predominance of intramural T-lymphocytes reflected a vasculitic process. The final diagnosis was cerebral vasculitis with subacute infarction. The patient died six days later. The autopsy findings were identical to the previous resection without evidence of systemic inflammation.

Safety Assessment of AAV2-hGDNF Administered Via Intracerebral Injection in Rats for Treatment of Parkinson's Disease.

Glial cell line-derived neurotrophic factor (GDNF) is a potent neuroprotective biologic in Parkinson's disease models. Adeno-associated viral vector serotype 2 (AAV2)-human GDNF safety was assessed in rats treated with a single intracerebral dose of vehicle, 6.8 × 108, 6.8 × 109, or 5.2 × 1010 vector genomes (vg)/dose followed by interim sacrifices on day 7, 31, 90, and 376. There were no treatment-related effects observed on food consumption, body weight, hematology, clinical chemistry, coagulation parameters, neurobehavioral parameters, organ weights, or serum GDNF and anti-GDNF antibody levels. Increased serum anti-AAV2 neutralizing antibody titers were observed in the 5.2 × 1010 vg/dose group. Histopathological lesions were observed at the injection site in the 6.8 × 109 vg/dose (day 7) and 5.2 × 1010 vg/dose groups (days 7 and 31) and consisted of gliosis, mononuclear perivascular cuffing, intranuclear inclusion bodies, and/or apoptosis on day 7 and mononuclear perivascular cuffing on day 31. GDNF immunostaining was observed in the injection site in all dose groups through day 376 indicating no detectable impacts of anti-AAV2 neutralizing antibody. There was no evidence of increased expression of calcitonin gene-related peptide or Swann cell hyperplasia in the cervical and lumbar spinal cord or medulla oblongata at the 5.2 × 1010 vg/dose level indicating lack of hyperplastic effects. In conclusion, no systemic toxicity was observed, and the local toxicity observed at the injection site appeared to be reversible demonstrating a promising safety profile of intracerebral AAV2-GDNF delivery. Furthermore, an intracerebral dose of 6.8 × 108 AAV2-GDNF vg/dose was considered to be a no observed adverse effect level in rats.

Cetacean Morbillivirus Infection in a Killer Whale (Orcinus orca) from Brazil

We provide pathological, immunohistochemical and molecular evidence of cetacean morbillivirus (CeMV) infection in a live-stranded adult female killer whale (Orcinus orca), which stranded alive in Espírito Santo State, Brazil, in 2014. Although attempts were made to release the animal, it stranded again and died. The main pathological findings were severe pulmonary oedema, pleural petechiation, multifocal, lymphoplasmacytic meningoencephalitis and leptomeningomyelitis with perivascular cuffing and gliosis, chronic lymphocytic bronchointerstitial pneumonia and multicentric lymph node and splenic lymphoid depletion. Other pathological findings were associated with the 'live-stranding stress response'. Immunohistochemical analysis revealed multifocal morbilliviral antigen in neurons and astrocytes, and in pneumocytes, histiocytes and leukocytes in the lung. CeMV was detected by a novel reverse transcriptase polymerase chain reaction method in the brain and kidney. Phylogenetic analysis of part of the morbillivirus phosphoprotein gene indicates that the virus is similar to the Guiana dolphin (Sotalia guianensis) morbillivirus strain, known to affect cetaceans along the coast of Brazil. To the authors' knowledge, this is the first report of morbillivirus disease in killer whales.

Modulation of experimental autoimmune encephalomyelitis through colonisation of the gut with Escherichia coli.

Multiple sclerosis (MS) is a neuro-inflammatory autoimmune disease of the central nervous system (CNS) that affects young adults. It is characterised by the development of demyelinating lesions and inflammation within the CNS. Although the causes of MS are still elusive, recent work using patient samples and experimental animal models has demonstrated a strong relationship between the gut microbiota and its contribution to CNS inflammation and MS. While there is no cure for MS, alteration of the gut microbiota composition through the use of probiotics is a very promising treatment. However, while most recent works have focused on the use of probiotics to modify pre-existing disease, little is known about its role in protecting from the establishment of MS. In this study, we determined whether colonisation with the probiotic bacterium Escherichia coli strain Nissle 1917 (EcN) could be used as a prophylactic strategy to prevent or alter the development of experimental autoimmune encephalomyelitis (EAE), a preclinical model of MS. We found that double gavage (two doses) of EcN before induction of EAE delayed disease onset and decreased disease severity. We also found that EcN-treated mice had decreased amounts of perivascular cuffing, CD4+ T cell infiltration into the CNS, together with significantly decreased absolute numbers of Th1 cells, and reduced activation of microglia. Although further studies are necessary to comprehend the exact protective mechanisms induced, our study supports a promising use of EcN as a probiotic for the prevention of MS.

Histopathological Differential Diagnosis of Meningoencephalitis in Cetaceans: Morbillivirus, Herpesvirus, Toxoplasma gondii, Brucella sp., and Nasitrema sp.

Infectious and inflammatory processes are among the most common causes of central nervous system involvement in stranded cetaceans. Meningitis and encephalitis are among the leading known natural causes of death in stranded cetaceans and may be caused by a wide range of pathogens. This study describes histopathological findings in post-mortem brain tissue specimens from stranded cetaceans associated with five relevant infectious agents: viruses [Cetacean Morbillivirus (CeMV) and Herpesvirus (HV); n = 29], bacteria (Brucella sp.; n = 7), protozoa (Toxoplasma gondii; n = 6), and helminths (Nasitrema sp.; n = 1). Aetiological diagnosis was established by molecular methods. Histopathologic evaluations of brain samples were performed in all the cases, and additional histochemical and/or immunohistochemical stains were carried out accordingly. Compared with those produced by other types of pathogens in our study, the characteristic features of viral meningoencephalitis (CeMV and HV) included the most severe and frequent presence of malacia, intranuclear, and/or intracytoplasmic inclusion bodies, neuronal necrosis and associated neuronophagia, syncytia and hemorrhages, predominantly in the cerebrum. The characteristic features of Brucella sp. meningoencephalitis included the most severe and frequent presence of meningitis, perivascular cuffing, cerebellitis, myelitis, polyradiculoneuritis, choroiditis, ventriculitis, vasculitis, and fibrinoid necrosis of vessels. The characteristic features of T. gondii meningoencephalitis included lymphocytic and granulomatous encephalitis, tissue cysts, microgliosis, and oedema. In the case of Nasitrema sp. infection, lesions are all that we describe since just one animal was available. The results of this study are expected to contribute, to a large extent, to a better understanding of brain-pathogen-associated lesions in cetaceans.

Simultaneous impact of atorvastatin and mesenchymal stem cells for glioblastoma multiform suppression in rat glioblastoma multiform model

Glioblastoma multiform (GBM) is known as an aggressive glial neoplasm. Recently incorporation of mesenchymal stem cells with anti-tumor drugs have been used due to lack of immunological responses and their easy accessibility. In this study, we have investigated the anti-proliferative and apoptotic activity of atorvastatin (Ator) in combination of mesenchymal stem cells (MSCs) on GBM cells in vitro and in vivo. The MSCs isolated from rats and characterized for their multi-potency features. The anti-proliferative and migration inhibition of Ator and MSCs were evaluated by MTT and scratch migration assays. The annexin/PI percentage and cell cycle arrest of treated C6 cells were evaluated until 72 h incubation. The animal model was established via injection of C6 cells in the brain of rats and subsequent injection of Ator each 3 days and single injection of MSCs until 12 days. The growth rate, migrational phenotype and cell cycle progression of C6 cells decreased and inhibited by the interplay of different factors in the presence of Ator and MSCs. The effect of Ator and MSCs on animal models displayed a significant reduction in tumor size and weight. Furthermore, histopathology evaluation proved low hypercellularity and mitosis index as well as mild invasive tumor cells for perivascular cuffing without pseudopalisading necrosis and small delicate vessels in Ator + MSCs condition. In summary, Ator and MSCs delivery to GBM model provides an effective strategy for targeted therapy of brain tumor.

Shuni virus-induced meningoencephalitis after experimental infection of cattle.

Shuni virus (SHUV), an insect-transmitted orthobunyavirus of the Simbu serogroup within the family Peribunyaviridae, may induce severe congenital malformations when naïve ruminants are infected during gestation. Only recently, another clinical presentation in cattle, namely neurological disease after postnatal infection, was reported. To characterize the course of the disease under experimental conditions and to confirm a causal relationship between the virus and the neurological disorders observed in the field, six calves each were experimentally inoculated (subcutaneously) with two different SHUV strains from both clinical presentations, that is encephalitis and congenital malformation, respectively. Subsequently, the animals were monitored clinically, virologically and serologically for three weeks. All animals inoculated with the 'encephalitis strain' SHUV 2162/16 developed viremia for three to four consecutive days, seroconverted, and five out of six animals showed elevated body temperature for up to three days. No further clinical signs such as neurological symptoms were observed in any of these animals. However, four out of six animals developed a non-suppurative meningoencephalitis, characterized by perivascular cuffing and glial nodule formation. Moreover, SHUV genome could be visualized in brain tissues of the infected animals by in situ hybridization. In contrast to the 'encephalitis SHUV strain', in animals subcutaneously inoculated with the strain isolated from a malformed newborn (SHUV 2504/3/14), which expressed a truncated non-structural protein NSs, a major virulence factor, no viremia or seroconversion, was observed, demonstrating an expected severe replication defect of this strain in vivo. The lack of viremia further indicates that virus variants evolving in malformed foetuses may represent attenuated artefacts as has been described for closely related viruses. As the neuropathogenicity of SHUV could be demonstrated under experimental conditions, this virus should be included in differential diagnosis for encephalitis in ruminants, and cattle represent a suitable animal model to study the pathogenesis of SHUV.

Rapid Intra-operative Diagnosis of CNS Lymphoma on Squash & Imprint Smears – A Boon for the Surgeon: Case Report

Primary CNS Lymphoma [PCNL] accounts for 3% of all brain tumors. Diagnosing PCNL on squash and imprint cytology is a challenge for the pathologist but a boon for the operating surgeon as the knowledge of the nature of the lesion determines the nature of surgery. We report a case of 59 years old immunocompetent male who presented with right hemiparesis and headache. MRI Brain was suggestive of Left frontal Glioma. Intra-operatively, squash cytology and imprint smears of tissue from the lesion showed scattered lymphoid cells which also exhibited perivascular cuffing. Cytology was suggestive of lymphoma with differential of an inflammatory pathology. Histopathological examination and positivity for LCA, CD 20, Bcl 2 and Mum 1 on IHC confirmed the diagnosis of Diffuse Large B Cell Lymphoma-Brain. It is important to distinguish lymphoma from glioma or metastasis in CNS because of totally different lines of management. Knowledge of empirical diagnosis during surgery can guide the surgeon to ensure that adequate tissue is excised so that sample for immunophenotyping or further tests is available.

Pathology of the outbreak of subgenotype 2.5 classical swine fever virus in northern Vietnam

Classical swine fever (CSF) is an endemic disease in southeastern Asia and is one of the most important swine diseases in Vietnam. This study was conducted to characterize the pathology of natural cases of CSF in northern Vietnam in 2018 and their genetic prevalence. A total of 10 representative pigs were collected from four provinces (Hung Yen, Ha Noi, Quang Ninh and Thai Binh) during five outbreaks and examined pathologically. The gross and histopathological findings showed the disease was expressed as the acute or the subacute to chronic form of CSF, depending on the age of the animals. The most consistently observed lesions associated with infection by the classical swine fever virus (CSFV) included lymphoid depletions in tonsils, lymph node and spleen; histiocytic hyperplasia in spleen; cerebral haemorrhage; perivascular cuffing in the brain; renal erythrodiapedesis; urothelial vacuolation and degeneration and interstitial pneumonia. The immunohistochemical findings showed a ubiquitous CSFV antigen mainly in the monocytes/macrophages and in the epithelial and endothelial cells in various organs. CSFV neurotropism was also found in the small neurons of the cerebrum and the ganglia of the myenteric plexus. Analysis of the full‐length envelope protein (E2) genome sequence showed that all strains were genetically clustered into subgenotype 2.5, sharing a nucleotide identity of 94.0%–100.00%. Based on the results of this study, the strain was categorized as a moderately virulent CSFV.