Immunohistochemical and molecular detection of equine herpesvirus 1 in Uruguay

Equine herpesvirus 1 (EHV-1) is a major cause of epidemic abortion, neonatal mortality, respiratory disease and neurological disorders in horses. In South America, the virus has been isolated in Brazil, Argentina and Colombia. In Chile pathological findings from one aborted foetus have been reported, and in Uruguay only serological data about EHV-1 activity have been found. Some pathological findings were reported in Uruguay several years ago, but these data have never been officially confirmed. The present work describes the relevant findings of a study of EHV-1 infections in the Uruguayan equine population using polymerase chain reaction (PCR) and histological and immunohistochemical analysis techniques. The sequence analysis of a portion of the glycoprotein C gene amplified by PCR confirmed EHV-1 activity. The real-time PCR revealed the association of the virus with the non-neuropathogenic genotype. This study describes for the first time the immunohistochemical and molecular detection of EHV-1 in Uruguay.Facultad de Ciencias Veterinaria

B-cell chronic lymphocytic leukaemia in an African lion (Panthera leo)

A 15-year-old female African lion (Panthera leo) maintained at the Zoological Garden of Pistoia, Tuscany (Italy), showed signs of malaise, dyspnoea, tremors, pale mucous membranes and ataxia for two days prior to death. Complete blood count showed anaemia, thrombocytopaenia and severe lymphocytosis. At autopsy, the most relevant finding was severe, generalised splenomegaly. Histologically, high numbers of neoplastic lymphocytes diffusely infiltrated the spleen, liver, heart, pancreas, kidney and lungs. The lymphoid cells were positive for CD79a and negative for CD3 on immunohistochemistry. The histologic features of the neoplastic cells and their immunophenotype are consistent with B-cell lymphocytes. Based on surface membrane co-expression of immunoglobulin M and immunoglobulin D in most neoplastic cells, which indicated that they were naïve, antigen-inexperienced and mature circulating resting B-cells, and consistent with their appearance as small lymphocytes with high nuclear-to-cytoplasmic ratios, a diagnosis of B-cell chronic lymphocytic leukaemia was made

Evaluation of Mud Worm (Polydora spp.) Infestation in Cupped (Crassostrea gigas) and Flat Oyster (Ostrea edulis) Broodstocks: Comparison between Magnetic Resonance Imaging and Computed Tomography

The Polichete worms of the genus Polydora are considered very destructive for oysters, excavating channels in their shell and inducing oysters to create mud blisters in response to the irritation, interfering with their physiology and ethology. The parasite also causes important economic damage for oyster farmers, as products with a high degree of infestation cannot be commercialized. The present study aims to evaluate whether two non-invasive advanced diagnostic techniques, computed tomography scans (CT) and magnetic resonance imaging (MRI), are suitable to show the alterations induced by this parasite on live Crassostrea gigas and Ostrea edulis oyster broodstocks. A CT scan is also able to identify small lesions in the shell during the first stage of infection. MRI allows for the visualization of the advanced status of the lesions when blisters occupy the inner surface of the shell and can impact the health status and the economic value of the mollusk. Both techniques resulted in satisfactory spatial resolution, and no motion artifacts were reported, thus enabling the authors to faithfully visualize in vivo the damage caused by the parasite

The diagnostic accuracy of the small fiber neuropathy symptoms inventory questionnaire (SFN-SIQ) for identifying pure small fiber neuropathy

A definite diagnosis of pure small fiber neuropathy (SFN) relies on specific diagnostic testing, such as skin biopsy, quantitative sensory testing (QST), and nociceptive evoked potentials, which require considerable resources that may not be widely available. Accordingly, diagnostic tools with easy implementation in non-specialist centers are warranted to identify patients who require second-level diagnostic tests. In this study, we aimed to test the accuracy of the Small Fiber Neuropathy Symptoms Inventory Questionnaire (SFN-SIQ) in diagnosing pure SFN. We enrolled 86 patients with suspected pure SFN. In these patients, we calculated the diagnostic accuracy of the SFN-SIQ using a combination of clinical examination, QST, and skin biopsy as a reference standard. We found that the SFN-SIQ showed an excellent ability to discriminate between patients with and without pure SFN, with 86% sensitivity and 70% specificity in the diagnosis of pure SFN. Our study providing the diagnostic yield of the SFN-SIQ for pure SFN diagnosis suggests that this questionnaire might be used to screen patients with suspected SFN and identify those requiring second-level diagnostic tests such as QST, skin biopsy, or nociceptive evoked potentials

GNAO1 encephalopathy: broadening the phenotype and evaluating treatment and outcome

OBJECTIVE: To describe better the motor phenotype, molecular genetic features, and clinical course of GNAO1-related disease. METHODS: We reviewed clinical information, video recordings, and neuroimaging of a newly identified cohort of 7 patients with de novo missense and splice site GNAO1 mutations, detected by next-generation sequencing techniques. RESULTS: Patients first presented in early childhood (median age of presentation 10 months, range 0-48 months), with a wide range of clinical symptoms ranging from severe motor and cognitive impairment with marked choreoathetosis, self-injurious behavior, and epileptic encephalopathy to a milder phenotype, featuring moderate developmental delay associated with complex stereotypies, mainly facial dyskinesia and mild epilepsy. Hyperkinetic movements were often exacerbated by specific triggers, such as voluntary movement, intercurrent illnesses, emotion, and high ambient temperature, leading to hospital admissions. Most patients were resistant to drug intervention, although tetrabenazine was effective in partially controlling dyskinesia for 2/7 patients. Emergency deep brain stimulation (DBS) was life saving in 1 patient, resulting in immediate clinical benefit with complete cessation of violent hyperkinetic movements. Five patients had well-controlled epilepsy and 1 had drug-resistant seizures. Structural brain abnormalities, including mild cerebral atrophy and corpus callosum dysgenesis, were evident in 5 patients. One patient had a diffuse astrocytoma (WHO grade II), surgically removed at age 16. CONCLUSIONS: Our findings support the causative role of GNAO1 mutations in an expanded spectrum of early-onset epilepsy and movement disorders, frequently exacerbated by specific triggers and at times associated with self-injurious behavior. Tetrabenazine and DBS were the most useful treatments for dyskinesia

Interferencia del Herpesvirus equino 1 (EHV-1) en la apoptosis inducida

La apoptosis es un tipo de muerte celular programada que puede ser desencadenada por múltiples factores, tanto internos como externos; dentro de estos últimos se encuentran las infecciones virales. Algunos alphaherpesvirus han desarrollado diversas estrategias para retardar o inhibir la muerte celular obteniendo, de esta manera, su propio beneficio al poder permanecer durante más tiempo en la célula. Hasta el momento no se ha identificado ningún mecanismo relacionado con la modulación de la muerte celular durante la infección con Herpesvirus equino tipo 1 (EHV-1). El objetivo del presente trabajo fue describir el efecto producido por la infección con EHV-1 sobre cultivos celulares inducidos a la muerte por apoptosis. La evaluación de la apoptosis se realizó mediante el reconocimiento de la fragmentación en escalera del ADN, la evaluación de la relación Anexina V/ioduro de propidio (IP) y la determinación del clivaje de la citoqueratina 18, utilizando técnicas de inmunofluorescencia. Los resultados indican una posible interferencia del EHV-1 con la muerte por apoptosis hacia la mitad de su ciclo de replicación, que se incrementa hacia el final del mismo.Apoptosis is a programmed cell death that can be triggered by many factors, both internal and external. Viral infections are included among the latter. Some alphaherpesvirus have developed several strategies to retard or inhibit cell death and thus the virus benefits itself by staying longer in the cell. So far, no mechanisms have been identified related to modulation of cell death during infection with equine herpesvirus type 1 (EHV-1). The aim of the present study was to describe the effect produced by the infection with EHV-1 on apoptosis-induced cell cultures. Assessment of apoptosis was performed by DNA laddering, the Annexin V/propidium iodide (PI) determination and the cytokeratin 18 cleavage analysis using immunofluorescence techniques. Results indicate a possible interference of EHV-1 with apoptotic cell death in the middle of its replication cycle, being increased by its end

Pain associated with COVID-19 vaccination is unrelated to skin biopsy abnormalities

Previous clinical observations raised the possibility that COVID-19 vaccination might trigger a small-fibre neuropathy.Objectives:In this uncontrolled observational study, we aimed to identify small fibre damage in patients complaining of generalized sensory symptoms and pain after COVID-19 vaccination.Methods:We collected clinical data, including a questionnaire for assessing autonomic symptoms (Composite Autonomic Symptom Score-31), and investigated quantitative sensory testing (QST) and skin biopsy in 15 prospectively enrolled patients with generalized sensory symptoms and pain after COVID-19 vaccination. Nine patients complaining of orthostatic intolerance also underwent cardiovascular autonomic tests.Results:We found that all patients experienced widespread pain, and most of them (11 of 15) had a fibromyalgia syndrome. All patients had normal skin biopsy findings, and in the 9 patients with orthostatic intolerance, cardiovascular autonomic tests showed normal findings. Nevertheless, 5 patients had cold and warm detection abnormalities at the QST investigation.Conclusions:In our study, most patients complaining of generalized sensory symptoms and pain after COVID-19 vaccination had clinical and diagnostic test findings compatible with a fibromyalgia syndrome. Although the abnormal QST findings we found in 5 patients might be compatible with a small-fibre neuropathy, they should be cautiously interpreted given the psychophysical characteristics of this diagnostic test. Further larger controlled studies are needed to define precisely the association between small fibre damage and COVID-19 vaccination

Detection of equine herpesvirus 1 genome 1B in Argentina

To determine the genomic variation of equine herpesviruses (EHVs) isolated in Argentina between 1979 and the first half of 2004, DNA sequences from all 69 strains isolated were analysed. Sixty strains were recovered from aborted fetuses, one from leucocyte-rich plasma from a horse with respiratory signs and eight from cases of neonatal disease. The DNA was extracted from rabbit kidney epithelial (RK13) cells infected with each strain and digested with three restriction endonucleases (BamHI, Bg/II and KpnI). Two strains could be differentiated using BamHI restriction and were assigned to the EHV-1 1B prototype group. Only one of these two strains was typed EHV-1 1B with Bg/II. DNA digestion with KpnI was ineffective. The results obtained in this study demonstrate that the EHV-1 1B genome has been present in Argentina since at least 1996. The finding of two strains with this electropherotype suggests that there is genomic heterogeneity among Argentinian isolates.Facultad de Ciencias Veterinaria

Analysis of the antibiotic resistance profiles in methicillin-sensitive s. Aureus pathotypes isolated on a commercial rabbit farm in Italy

The breeding of meat rabbits is an important sector in the livestock industry in Italy. The focus of this study was to describe the antibiotic resistance profile distribution among the Methicillin-sensitive Staphylococcus aureus isolated in a rabbit farm. From 400 animals of different ages and three farm workers, 96 randomly selected strains isolated from various anatomical sites and lesions were analysed. According to spa typing and the resistance profiles towards veterinary and human antibiotics, 26 pathotypes were identified. The highest resistance was observed against Tetracyclines (92.3%) and Macrolides (80.8%), while almost all were susceptible to Penicillins, according to the limited use of β-lactams on the farm. In total, 92.3% of pathotypes were multidrug resistant (MDRs). Two MDR pathotypes belonging to the t2802 spa type were isolated from both farmers and rabbits. Age categories harboured significantly different pathotypes (p = 0.019), while no association was found between pathotypes and lesions (p = 0.128) or sampling sites (p = 0.491). The antibiotic resistance was observed to increase with the time spent in the farm environment (age category). The selective pressure exerted by antibiotic use acted by giving advantage to more resistant strains rather than by lowering susceptibility to various drug categories within strains

Greenlight laser™ photovaporization versus transurethral resection of the prostate: A systematic review and meta-analysis

none9GreenLight laser™ photovaporization of the prostate (GLL-PVP) has become a valid alternative to traditional transurethral resection of the prostate (TURP) in men requiring surgery for benign prostatic hyperplasia. We aimed to review systematically the safety and efficacy of studies comparing GLL PVP and TURP in the medium-term. A comprehensive literature search was performed. Twelve studies were identified for meta-analysis. Meta-analyses showed a longer postoperative catheterization time (risk ratio (RR): 1.12, 95% CI:1.09–1.14, p<0.00001) and length of stay (RR: 1.16, 95% CI:1.12–1.19, p<0.00001) in the TURP group; higher risk of transfusion in the TURP group (RR: 6.51, 95% CI: 2,90–14,64 p<0.00001); no difference in the risk of urinary tract infections (RR: 0.83, 95% CI: 0.58–1.18, p=0.30) and transient re-catheterization (RR: 1.11, 95% CI: 0.76–1.60, p=0.60). Regarding reoperation rate, no difference was found in term of postoperative urethral stricture (RR: 1.13, 95% CI: 0.73–1.75, p=0.59) and bladder neck contracture (RR: 0.66, 95% CI: 0.31–1.40, p=0.28). A significantly higher incidence in reoperation for persistent/regrowth adenoma was present in the GLLL-PVP (RR: 0.64, 95% CI: 0.41–0.99, p=0.05). Data at 2-year follow-up showed significant better post-voiding residual (PVR) (MD:-1.42, 95% CI:-2.01,-0.82, p<0.00001) and International Prostate Symptom Score (IPSS) (MD:-0.35, 95% CI:-0.50,-0.20, p<0.00001) after TURP. No difference was found in the mean PVR at 2 years after TURP, in the mean maximum flow rate (Qmax) (MD: 0.30, 95% CI:-0.02–0.61, p=0.06) and quality of life QoL score (MD: 0.05, 95% CI:-0.02–0.42, p=0.13). At 5-year follow-up, data showed better IPSS (MD:-1.70, 95% CI:-2.45,-0.95, p<0.00001), QoL scores (MD:-0.35, 95% CI:-0.69,-0.02, p=0.04) and Qmax (MD: 3.29, 95% CI: 0.19–6.38, p=0.04) after TURP. Data of PVR showed no significant difference (MD:-11.54, 95% CI:-29.55–6.46, p=0.21). In conclusion, our analysis shows that GLL-PVP is a safer and more efficacious procedure than standard TURP in the early and medium-term. However, in the long term period GLL-PVP showed a higher incidence of reoperation rate due to incomplete vaporization/regrowth of prostatic adenoma.openCastellani D.; Pirola G.M.; Rubilotta E.; Gubbiotti M.; Scarcella S.; Maggi M.; Gauhar V.; Teoh J.Y.-C.; Galosi A.B.Castellani, D.; Pirola, G. M.; Rubilotta, E.; Gubbiotti, M.; Scarcella, S.; Maggi, M.; Gauhar, V.; Teoh, J. Y. -C.; Galosi, A. B