Effects of Autohemotherapy on Hematologic Parameters and Morphology of Canine Oral Papillomatosis

Background: Several treatments for oral papillomatosis in dogs, with satisfactory results, have already been described in the literature. However, there is need for further studies regarding remission time, association and exposure to drugs, side effects, cost-effectiveness and reproducibility of results. Autohemotherapy (AHT) is a therapeutic technique increasingly used in veterinary medicine that has been providing satisfactory clinical results to treat several pathologies. The procedure consists of removing blood by intravenous puncture that is immediately re-administered via intramuscular, subcutaneous, intravenous or intradermal. Although the empirical isolated use via intramuscular reached excellent results in routine medical treatment of dogs and since few studies and protocols have been described for this species, many times the treatment is based on the experience with the bovine papillomatosis. To this end, this report describes a case of canine oral papillomatosis treated with autohemotherapy (AHT), emphasizing the dosage and clinical evolution of the case.Case: A one-year-old mongrel dog rescued from the street eight months earlier was treated. The animal was thin resulting from the small food amount ingested due to feeding difficulty, infested with ticks, had bad breath and few small warts on the lips, which, after five months, grew and multiplied, and were distributed throughout the oral cavity. The animal clinical history led to papillomatosis diagnosis, which was confirmed by cytological and histopathological exams. The AHT treatment consisted of taking 5 mL of whole blood without EDTA and immediately injecting this volume intramuscularly, every four days until the total remission of the clinical symptoms. Every four days, the papilloma fall off, and involution processes were recorded through a detailed macroscopic analysis of the warts (quantity, consistency, color and size), as well as hematological evaluations (blood count, hemoparasite screening and total plasm protein), which were performed before each AHT treatment until the complete clinical resolution of the disease (24 days).Discussion: The dose adopted in this autohemotherapy study was clinically effective since the papillomas started to fall off after four days of therapy, with total resolution in 24 days. Therefore, it can be concluded that the 5 mL AHT injected intramuscularly every four days reduced treatment time and decreased therapeutic costs. There are few studies evaluating more accurately the hematological aspects related to the therapeutic use of autohemotherapy to treat canine oral papillomatosis, the existing reports are restricted to the initial measurement as an evaluation of the general condition of the patient and/or diagnosis of possible basal diseases. In this study, no changes were observed in the blood count during the entire therapeutic process, demonstrating that the AHT did not interfere in the red series dynamics of the treated animal. Additionally, thrombocytosis followed by thrombocytopenia exhibited a correlation with the macroscopic appearance of warts [enlargement and modified color, texture, shape and consistency]. This observation is possibly related to a platelet mobilization toward the site of action [warts], so that there was a modulation in the immune response, amplifying the inflammatory process and tissue repair, which resulted in the fall off and involution of the papillomas

Comparative molecular cytogenetics in Melipona Illiger species (Hymenoptera, Apidae)

Cytogenetic studies in Melipona are scarce with only 24 species analyzed cytogenetically. Of these, six species had the rDNA sites physically mapped and characterized by Fluorescent in situ Hybridization (fish). The aim of this study was to perform karyotype analyzes on Melipona species from different regions of Brazil, with a greater sampling representative of the Amazonian fauna and using conventional, fluorochrome staining and FISH with heterologous rDNA probes. The predominant chromosome number was 2n = 18, however, the subspecies M. seminigra abunensis and M. s. pernigra showed 2n = 22 chromosomes. The karyotypes were symmetrical, however M. bicolor, M. quadrifasciata, M. flavolineata, M. fuscopilosa, M. nebulosa presented the first pair heteromorphic in length. CMA3+ blocks also exhibited heteromorphism of size and in almost all cases coincided with rDNA sites, except for M. crinita and M. nebulosa, which presented additional non-coincident CMA3+ blocks. The CMA/ rDNA sites were terminal and interstitial in species with high heterochromatic content, and pericentromeric in those species with low heterochromatic content. In addition to pointing out cytogenetic features of cytotaxonomic importance, the reorganization of the genome in Melipona is discussed

Role of bolA and rpoS genes in biofilm formation and adherence pattern by Escherichia coli K-12 MG1655 on polypropylene, stainless steel, and silicone surfaces

Escherichia coli has developed sophisticated means to sense, respond, and adapt in stressed environment. It has served as a model organism for studies in molecular genetics and physiology since the 1960s. Stress response genes are induced whenever a cell needs to adapt and survive under unfavorable growth conditions. Two of the possible important genes are rpoS and bolA. The rpoS gene has been known as the alternative sigma (σ) factor, which controls the expression of a large number of genes, which are involved in responses to various stress factors as well as transition to stationary phase from exponential form of growth. Morphogene bolA response to stressed environment leads to round morphology of E. coli cells, but little is known about its involvement in biofilms and its development or maintenance. This study has been undertaken to address the adherence pattern and formation of biofilms by E. coli on stainless steel, polypropylene, and silicone surfaces after 24 h of growth at 37 °C. Scanning electron microscopy was used for direct examination of the cell attachment and biofilm formation on various surfaces and it was found that, in the presence of bolA, E. coli cells were able to attach to the stainless steel and silicone very well. By contrast, polypropylene surface was not found to be attractive for E. coli cells. This indicates that bolA responded and can play a major role in the presence and absence of rpoS in cell attachment

Cryptococcal Sialoadenitis in a Dog

Background: Salivary gland diseases in dogs have an overall incidence of 0.3%. Sialocele and sialoadenitis are the most common injuries and are usually caused by infectious diseases, especially of bacterial origin. Currently, Cryptococcus neoformans has not been registered as the etiological agent since fungal participation is unusual. This case report describes a cryptococcal sialoadenitis in a dog.Case: A 1.9-year-old male poodle had a swelling of the right submandibular region, combined with dry cough and snoring that persisted for about 30 days. The dog often walked in the town square, which has pigeons, and was seen eating chicken droppings. The dog was diagnosed with sialoadenitis in the right submandibular salivary gland. The following laboratory tests were performed cytology, fungal culture, blood count, search for hemoparasites, total plasma protein (ppt), alanine aminotransferase dosage (ALT), alkaline phosphatase (ALP), urea (U), creatinine (CREA), Albumin (ALB), total protein (TP), globulin (GLOB) and amylase (AML). In addition, the dog underwent cervical, thoracic and abdominal ultrasound, and cervical x-rays. The cytology results showed fungal sialoadenitis caused by Cryptococcus neoformans. The blood testresults evidenced thrombocytopenia and increased ppt, TP, GLOB, and AML. The lungs and heart x-rays results showed no alterations. However, the proximal third of the trachea was displaced ventrally. The abdominal area had no changes, but hypertrophy of the salivary gland was observed, with irregular contour, heterogeneous echotexture, and multiple cavitaryareas flled with intraluminal fluid of cellular appearance. The ketoconazole (5 mg/kg/BID/20 days) treatment prescribed led to drug-induced hepatitis. The new treatment with Itraconazole (10 mg/kg/SID/90 days) resulted in complete clinical resolution without any side effect or relapse one year after the treatment.Discussion: Although poodle is not commonly associated with the epidemiology of the disease, this breed seems to be susceptible, as well. The contact with pigeons during the walks was likely the source of infection by C. neoformans, but the ingestion of chicken droppings cannot be discarded. The coughing and snoring resulted from the pharyngeal and tracheal compression, with subsequent activation of the reflex. The clinical and laboratory fndings suggest a new infection route by the etiological agent, possibly the opening of the salivary duct located in the sublingual carbuncle. Biopsy or ablation of the affected salivary gland was not necessary, reducing the risk of sepsis and/or death. Hyperamylasemia indicates that AML can be used as a complementary tool to diagnose sialoadenitis. Itraconazole administered for 90 days resulted incomplete healing while no side effects or relapse was observed one year after the treatment. Cryptococcus neoformans can affect the submandibular salivary gland and, therefore, cryptococcosis should enter the differential diagnosis list of canine sialoadenitis and sialocele. Itraconazole has been shown to be effective to treat a dog with sialoadenitis caused by Cryptococcus neoformans.Keywords: sialoadenitis, sialocele, cryptococcosis, salivary gland, dog

Multicentric Squamous Cell Carcinoma with the Involvement of the Ocular Annexes in a Dog

Background: The cutaneous squamous cell carcinoma (cSCC) is considered to be a frequent neoplasm in dogs, however, its origin in ocular annexes, especially in relation to the conjuctival location, is a rare finding in dogs. Therefore, it was aimed to report the occurrence of a multicentric SCC, with the involvement of ocular annexes in a dog, emphasizing its clinical characteristics and histopathological findings.Case: A 6-year-old non-castrated white-coated Pitbull dog was attended, with a history of increased volume and bloody secretion in the left eye, with an evolution of approximately six months. By means of general physical examination, ulcerated lesions in the foreskin and scrotum were found. During the ophthalmologic examination was identified an extensive and irregular exophytic mass, of a reddish color and with a cauliflower-like appearance, located in the inferior bulbar conjunctiva and third eyelid of the left eye, accompanied by a large quantity of piosanguinolenta secretion, mainly during manipulation. Other alterations were observed, such as, meibomitis, conjuctival hyperemia, hypopyon, corneal edema and loss of sight. In the right eye, the only alteration found was conjunctival hyperemia. The hemograma revealed discreet anemia; the serum biochemical profile was inside the normal range and there was no evidence of metastasis in the imaging examinations. The animal was submitted to the incisional biopsy of the lesions for histopathological analysis, which revealed a proliferation of neoplastic epithelial cells, highly pleomorphic, composed of eosinophilic cytoplasm, which varied from scarce to moderate, of indistinct borders, with a large nucleus and loose chromatin and large and evident nucleolus, compatible with SCC, enabling, also, the classification as multicentric due to the multiple localizations. Additionally, associated to the conjunctival tumor, there was necrosis and mixed inflammatory infiltration; in the scrotum and conjunctiva, the cells presented more accentuated pleomorphism, with the presence of dyskeratosis and little formation of keratin pearls; however in the prepuce, there was abundant formation of keratin pearls in the midst of the tumor. In the immunohistochemical analysis, the neoplastic cells demonstrated strong and uniform cytoplasmic immunoreactivity for pancytokeratin. It was recommended the exenteration of the left eye followed by the introduction of acrylic resin intraorbital implant, together with the resection of the neoplasm from the scrotum and foreskin, associated with cryotherapy. However, the owner was reluctant to the proposed treatment and opted for the euthanasia of the animal, not consenting to the performance of the necropsy.Discussion: The etiological factors related to the development of the SCCs, especially concerning those of the ocular and periocular region, in dogs and cats, are still not well defined. However, the overexposure to the ultraviolet radiation has been pointed as the main etiological factor, especially in tropical and high-altitude regions. Indeed, the characteristics of the region in which the animal resided, associated to its way of life and its phenotypical characteristics suggested that the chronic exposure to ultraviolet radiation would be the most plausible cause related to the emergence of the multicentric SCC of this case. Thus, it is suggested that, while the physiopathology of the neoplasm has still yet not been elucidated, it must be avoided that the dogs, with these characteristics, expose themselves too much to solar radiation.

New occurrence of B chromosomes in Partamonahelleri (Friese, 1900) (Hymenoptera, Meliponini)

Cytogenetic analyses of the stingless bee Partamona helleri collected in the state of Bahia, Northeast Brazil revealed the chromosome numbers n = 18 in the haploid males and 2n = 35 in the diploid females. All karyotypes displayed one large acrocentric B chromosome, which differs from the minute B chromosomes previously described in the populations from southeastern Brazil. Giemsa staining, C-banding and DAPI/CMA3 fluorochrome staining also revealed a remarkable interpopulational divergence regarding both the regular karyotype and the B chromosomes. The B chromosomes found in the samples from Jequié, Bahia, were entirely heterochromatic, while those found in Cravolândia, Bahia, displayed a euchromatic portion at the telomeric end of the long arm. CMA 3 labeling sites varied from seven to eight between the two localities in Bahia, due to the presence of an extra GC-rich block in the karyotype of the samples from Jequié. This is the first report of a large B chromosome in P. helleri and reveals the occurrence of a geographic differentiation within this species

Persistent symptoms and decreased health-related quality of life after symptomatic pediatric COVID-19: A prospective study in a Latin American tertiary hospital

OBJECTIVES: To prospectively evaluate demographic, anthropometric and health-related quality of life (HRQoL) in pediatric patients with laboratory-confirmed coronavirus disease 2019 (COVID-19) METHODS: This was a longitudinal observational study of surviving pediatric post-COVID-19 patients (n=53) and pediatric subjects without laboratory-confirmed COVID-19 included as controls (n=52) was performed. RESULTS: The median duration between COVID-19 diagnosis (n=53) and follow-up was 4.4 months (0.8-10.7). Twenty-three of 53 (43%) patients reported at least one persistent symptom at the longitudinal follow-up visit and 12/53 (23%) had long COVID-19, with at least one symptom lasting for >12 weeks. The most frequently reported symptoms at the longitudinal follow-up visit were headache (19%), severe recurrent headache (9%), tiredness (9%), dyspnea (8%), and concentration difficulty (4%). At the longitudinal follow-up visit, the frequencies of anemia (11% versus 0%, p=0.030), lymphopenia (42% versus 18%, p=0.020), C-reactive protein level of >30 mg/L (35% versus 0%, p=0.0001), and D-dimer level of >1000 ng/mL (43% versus 6%, p=0.0004) significantly reduced compared with baseline values. Chest X-ray abnormalities (11% versus 2%, p=0.178) and cardiac alterations on echocardiogram (33% versus 22%, p=0.462) were similar at both visits. Comparison of characteristic data between patients with COVID-19 at the longitudinal follow-up visit and controls showed similar age (p=0.962), proportion of male sex (p=0.907), ethnicity (p=0.566), family minimum monthly wage (p=0.664), body mass index (p=0.601), and pediatric pre-existing chronic conditions (p=1.000). The Pediatric Quality of Live Inventory 4.0 scores, median physical score (69 [0-100] versus 81 [34-100], p=0.012), and school score (60 [15-100] versus 70 [15-95], p=0.028) were significantly lower in pediatric patients with COVID-19 at the longitudinal follow-up visit than in controls. CONCLUSIONS: Pediatric patients with COVID-19 showed a longitudinal impact on HRQoL parameters, particularly in physical/school domains, reinforcing the need for a prospective multidisciplinary approach for these patients. These data highlight the importance of closer monitoring of children and adolescents by the clinical team after COVID-19

BiofOmics: A Web Platform for the Systematic and Standardized Collection of High-Throughput Biofilm Data

Background: Consortia of microorganisms, commonly known as biofilms, are attracting much attention from the scientific community due to their impact in human activity. As biofilm research grows to be a data-intensive discipline, the need for suitable bioinformatics approaches becomes compelling to manage and validate individual experiments, and also execute inter-laboratory large-scale comparisons. However, biofilm data is widespread across ad hoc, non-standardized individual files and, thus, data interchange among researchers, or any attempt of cross-laboratory experimentation or analysis, is hardly possible or even attempted. Methodology/Principal findings This paper presents BiofOmics, the first publicly accessible Web platform specialized in the management and analysis of data derived from biofilm high-throughput studies. The aim is to promote data interchange across laboratories, implementing collaborative experiments, and enable the development of bioinformatics tools in support of the processing and analysis of the increasing volumes of experimental biofilm data that are being generated. BiofOmics data deposition facility enforces data structuring and standardization, supported by controlled vocabulary. Researchers are responsible for the description of the experiments, their results and conclusions. BiofOmics curators interact with submitters only to enforce data structuring and the use of controlled vocabulary. Then, BiofOmics search facility makes publicly available the profile and data associated with a submitted study so that any researcher can profit from these standardization efforts to compare similar studies, generate new hypotheses to be tested or even extend the conditions experimented in the study. Significance BiofOmics novelty lays on its support to standardized data deposition, the availability of computerizable data files and the free-of-charge dissemination of biofilm studies across the community. Hopefully, this will open promising research possibilities, namely: the comparison of results between different laboratories, the reproducibility of methods within and between laboratories, and the development of guidelines and standardized protocols for biofilm formation devices and analytical methods.The financial support from the Institute of Biotechnology and Bioengineering - Center of Biological Engineering (IBB-CEB), Fundacao para a Ciencia e Tecnologia (FCT) and European Community fund FEDER (Program COMPETE), project PTDC/SAU-ESA/646091/2006/FCOMP-01-0124-FEDER-007480 and PhD grant of Idalina Machado (SFRH/BD/31065/2006) are gratefully acknowledged. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript