Stafne's bone defect: a systematic review.

BACKGROUND: This systematic review integrated the available data published in the literature on Stafne's bone defect (SBD), considering the clinical, imaging and histopathological results. MATERIAL AND METHODS: An electronic search was undertaken in six databases. Eligibility criteria were: articles in English, Spanish, and Portuguese describing case reports or case series of SBD, reported up to September/2021. Risk of bias was assessed using the Joanna Briggs Institute tool. RESULTS: A total of 98 articles were retrieved, involving 465 individuals with SBD and were included for quantitative analysis. Mean age was 52.78 years (range: 11-89 years), with male predilection (n=374/80.85%). Radiographs were the most frequent imagiological exams (n=298/64.09%), followed by computed tomography (n=98/21.08%). SBD was more prevalent in the posterior mandible (n=361/93.77%) as a hypodense radiolucent lesion (n=250/77.40%). Mean size was 1.58 cm (range: 0.3-.8.0 cm). Two-hundred-and-two lesions (97.37%) were unilocular and 126 (91.97%) were classified as well-defined. Clinical symptoms were reported in 73 cases, while 68 cases (93.15%) were asymptomatic. Only 34 cases (12.32%) were submitted to histopathological examination. Mean follow-up time was 26.42 ±25.39 months. CONCLUSIONS: SBD is more frequent in male patients in the fifth and sixth decade of life. Classic SBD is radiographically characterized as a single, unilocular and well-defined lesion in the posterior region of the jaw with a radiolucent/hypodense appearance.

Is teriparatide therapy effective for medication-related osteonecrosis of the jaw? A systematic review and meta-analysis.

Considering the lack of studies determining the real TPTD efficacy in individuals who develop MRONJ, our objective was to combine the available data on MRONJ cases treated with TPTD. The findings demonstrated that TPTD in combination with another therapy, especially antibiotic therapy, can be considered an effective protocol for MRONJ. PURPOSE: To integrate the data published on the effect of teriparatide (TPTD) therapy on cases of medication-related osteonecrosis of the jaws (MRONJ) into a comprehensive analysis of clinical features. METHODS: An electronic search was undertaken in six databases. Descriptive analyses of clinicodemographic data of MRONJ were carried out. Poisson regression was also run to evaluate predictors of total resolution of MRONJ treated with TPTD. RESULTS: Twenty-six publications comprising 111 cases were included. Most reported cases affected female individuals (82.0%) with a mean age of 76.54 years. Osteoporosis (76.5%) represented the main reason for using antiresorptive drugs, with bisphosphonates (98.1%) as the most frequently reported. Comorbidities were commonly present. The most related trigger factor of MRONJ was dental extraction (61.7%). Mandible (75.8%) was the most commonly affected site, with a mean evolution time of 5 months. MRONJ stage 2 (61.3%) was the most prevalent. Regarding TPTD treatment, in 45.1% cases, TPTD was used alone, with the total resolution being observed in 59.5% of the individuals. Associated therapy (54.9%) included surgery, antibiotic therapy, and laser therapy. Mean follow-up was 8.7 months. Poisson regression demonstrated that individuals with MRONJ stage 1 were 1.21 times more likely to present total resolution of osteonecrosis than individuals with MRONJ stage 3 (CI = 1.02-1.43; p < 0.023). Individuals who had undergone treatment with TPTD in association with another therapeutic modality were 1.21 times more likely to present total resolution of osteonecrosis than those who had undergone treatment with TPTD alone (CI = 1.40-1.39; p < 0.010). CONCLUSION: TPTD in combination with another therapy, especially antibiotic therapy, should be considered an effective therapeutic modality for MRONJ.

Histological and immunohistochemical evaluation of two cell therapy protocols in equine suspensory ligament repair / Avaliação histológica e imunoistoquímica do reparo do ligamento suspensório equino tratado com dois protocolos de terapia celular

This study aimed to histologically evaluate the quality of tissue repair in equine suspensory ligament treated with two cell therapy protocols. All four limbs of six animals were operated simultaneously to remove a fragment in each ligament using a skin biopsy punch. Two days later, intralesional injections were performed using bone marrow mononuclear fraction (BM group), cultivated cells derived from adipose tissue (AT group), saline (positive control group), or no treatment (negative control group), in such way that each horse received all treatments. After sixty days biopsies were performed for histological analysis (H & E, Masson's trichrome and picrosirius red) and immunohistochemistry analysis (collagen type III). Histological findings (H & E and Masson's trichrome), birefringence intensity (through picrosirius) and collagen type III expression (through immunohistochemistry) were analyzed. Samples from treated groups had better birefringence intensity (P=0.007) and fiber alignment scores were superior compared to controls, though not statistically significant (P=0.08). Presence of inflammatory cells and intense staining for collagen type III occurred in all groups demonstrating an active healing process. In conclusion, both protocols resulted in improvement of tissue repair indicating their potential to be used as an adjuvant treatment of equine suspensory ligament disorders.(AU)

Este estudo teve como objetivo a avaliação histológica e imunoistoquímica do reparo do ligamento suspensório equino tratado com dois protocolos de terapia celular. Os quatro membros dos seis animais do experimento foram submetidos a procedimento cirúrgico em que um fragmento de cada ligamento foi retirado, utilizando-se punch de biópsia. Dois dias após o procedimento, aplicações intralesionais foram realizadas, por meio de aspirado de medula óssea (bone marrow-BM), células mesenquimais derivadas de tecido adiposo (adipose tissue-AT), solução salina (positive control group-PC) ou controle (negative control-NC). Após 60 dias, biópsias foram retiradas da região de reparo dos ligamentos e foram submetidas à análise histológica (HE, tricrômio de Masson, picrosírius red) e imunoistoquímica (colágeno tipo III). Diferentes variáveis histológicas (HE e tricrômio de Masson), a intensidade de birrefringência das fibras colágenas (picrosírius red) e a expressão de colágeno tipo III foram avaliadas. Os grupos tratados apresentaram maior birrefringência (P=0,007) e alinhamento de fibras (P=0,08) comparados ao controle, para o qual o resultado não se mostrou estatisticamente significativo. Achados histológicos e imunoistoquímicos demonstraram um processo ativo de reparo tecidual em todos os grupos. Concluiu-se que os dois protocolos de terapia celular apresentaram melhora no reparo tecidual, demonstrando potencial terapêutico adjuvante no tratamento de afecções do ligamento suspensório equino.(AU)

Histological and immunohistochemical evaluation of two cell therapy protocols in equine suspensory ligament repair / Avaliação histológica e imunoistoquímica do reparo do ligamento suspensório equino tratado com dois protocolos de terapia celular

This study aimed to histologically evaluate the quality of tissue repair in equine suspensory ligament treated with two cell therapy protocols. All four limbs of six animals were operated simultaneously to remove a fragment in each ligament using a skin biopsy punch. Two days later, intralesional injections were performed using bone marrow mononuclear fraction (BM group), cultivated cells derived from adipose tissue (AT group), saline (positive control group), or no treatment (negative control group), in such way that each horse received all treatments. After sixty days biopsies were performed for histological analysis (H & E, Masson's trichrome and picrosirius red) and immunohistochemistry analysis (collagen type III). Histological findings (H & E and Masson's trichrome), birefringence intensity (through picrosirius) and collagen type III expression (through immunohistochemistry) were analyzed. Samples from treated groups had better birefringence intensity (P=0.007) and fiber alignment scores were superior compared to controls, though not statistically significant (P=0.08). Presence of inflammatory cells and intense staining for collagen type III occurred in all groups demonstrating an active healing process. In conclusion, both protocols resulted in improvement of tissue repair indicating their potential to be used as an adjuvant treatment of equine suspensory ligament disorders.(AU)

Este estudo teve como objetivo a avaliação histológica e imunoistoquímica do reparo do ligamento suspensório equino tratado com dois protocolos de terapia celular. Os quatro membros dos seis animais do experimento foram submetidos a procedimento cirúrgico em que um fragmento de cada ligamento foi retirado, utilizando-se punch de biópsia. Dois dias após o procedimento, aplicações intralesionais foram realizadas, por meio de aspirado de medula óssea (bone marrow-BM), células mesenquimais derivadas de tecido adiposo (adipose tissue-AT), solução salina (positive control group-PC) ou controle (negative control-NC). Após 60 dias, biópsias foram retiradas da região de reparo dos ligamentos e foram submetidas à análise histológica (HE, tricrômio de Masson, picrosírius red) e imunoistoquímica (colágeno tipo III). Diferentes variáveis histológicas (HE e tricrômio de Masson), a intensidade de birrefringência das fibras colágenas (picrosírius red) e a expressão de colágeno tipo III foram avaliadas. Os grupos tratados apresentaram maior birrefringência (P=0,007) e alinhamento de fibras (P=0,08) comparados ao controle, para o qual o resultado não se mostrou estatisticamente significativo. Achados histológicos e imunoistoquímicos demonstraram um processo ativo de reparo tecidual em todos os grupos. Concluiu-se que os dois protocolos de terapia celular apresentaram melhora no reparo tecidual, demonstrando potencial terapêutico adjuvante no tratamento de afecções do ligamento suspensório equino.(AU)

Forty cases of acquired oral syphilis and a review of the literature.

The aim of this study was to describe 40 cases of acquired oral syphilis (AOS) and to discuss the distribution of demographic characteristics, clinical features, and differential diagnosis of the disease. A retrospective study was conducted covering a 17-year period at a single institution in southern Brazil. Moreover, a literature review was performed through a search of the PubMed database for articles on AOS published between 1955 and March 2018. Data were analyzed descriptively. The predominant group within the case series was male patients in their twenties. The vast majority of cases (92.5%) were in the secondary stage of the disease. The lips were the most commonly affected site, with greyish-white mucous patches and reddish ulcers. In the literature review, the largest number of reported cases came from North America. Male patients in the third and fourth decades of life were most affected. AOS occurred more commonly as mucous patches and ulcers on the tongue and palate. Similarities regarding the distribution by sex, age, and anatomical location were found in the present study when compared to cases reported elsewhere. Clinicians, oral pathologists, and maxillofacial surgeons should familiarize themselves with the variable spectrum of signs and symptoms of AOS in their clinical practice to improve diagnosis and management.

Retrospective analysis of jaw biopsies in young adults. A study of 1599 cases in Southern Brazil.

BACKGROUND: To evaluate the prevalence and the characteristics of jaw lesions diagnosed in young adults aged 20 to 30 years in a southern Brazil reference center, over a period of 25 years. And to analyze the concordance between clinical and histological diagnosis. MATERIAL AND METHODS: In this cross-sectional retrospective study, the biopsies files from this center were retrieved and data regarding sex, age, bone localization, clinical and histological diagnosis were collected. The histological diagnosis were grouped into the categories Cystic lesions of odontogenic origin, Periapical inflammation, Odontogenic tumors, Bone diseases, Health tissue and Nonspecific diagnostic. Absolute and relative frequencies were estimated with descriptive analysis. The agreement between clinical and histological diagnosis was measured through Kappa statistic. RESULTS: A total of 18,181 histopathological analysis were performed during the period of the study, registering 1,599 jaw lesions in young adults. The average age of individuals was 24,59 years (SD 3,1). Nine hundred ninety-one (62%) lesions were found in females and 608 (38%) in males. More than half of pathologies were cystic lesions of odontogenic origin (822/51.4%), followed by periapical inflammation (282/17.6%). Regarding the site of lesions, more than half occurred in posterior mandible (877/54.8%), followed by posterior maxilla (339/21.2%). The most frequent entities were periapical cyst, chronic periapical granuloma, dental follicle and paradental cyst, corresponding to a total of 1,202 (75.2%) evaluated cases. In relation to the analysis of concordance between clinical and histological diagnosis the general Kappa index was 0.5, which is considered moderate. Finally, the findings confirm data from literature about the most frequent jaw pathologies in young adults and serve as aid for preventive measures of some entities. Additionally, they can improve the formulation of differential diagnosis and the patient management.

Steatosis caused by experimental periodontitis is reversible after removal of ligature in rats.

BACKGROUND AND OBJECTIVE: Periodontitis may promote harmful systemic effects such as changes in hepatic tissues. The purpose of this study was to investigate whether the steatosis and oxidative stress caused by experimental periodontitis are reversible in the liver. MATERIAL AND METHODS: Twenty-four rats were divided into three groups: control, periodontitis and P20-20 (20 days with experimental periodontitis and 20 days without experimental periodontitis, to verify the reversibility of hepatic injuries). The following parameters were assessed: gingival bleeding index, probing pocket depth, myeloperoxidase activity, alveolar bone loss for periodontal tissues; liver weights, histopathological scores for steatosis, inflammation and necrosis in liver; glutathione, malondialdehyde, total cholesterol and triglyceride concentrations in hepatic tissues; and blood levels of aspartate aminotransferase, alanine aminotransferase, albumin, gamma-glutaryl transferase, total cholesterol and random glucose. RESULTS: Gingival bleeding index, probing pocket depth, myeloperoxidase and alveolar bone loss parameters demonstrated the development of periodontitis. There was a significant reduction in the steatosis score of animals from the P20-20 group when compared with the periodontitis group. P20-20 group presented significantly higher glutathione (11 times) and lower malondialdehyde (nearly 23%), total cholesterol (both in blood and hepatic tissue) and triglyceride concentrations compared with the periodontitis group. For levels of aspartate aminotransferase, alanine aminotransferase, albumin, gamma-glutaryl transferase and random glucose, a significant difference between the groups was not observed. CONCLUSION: Our results demonstrate that the microvesicular steatosis caused by periodontitis in rats is reversible after removal of the ligature, which is associated with the increase in oxidative stress and lipid peroxidation in the liver.

Simple and aneurysmal bone cyst: Aspects of jaw pseudocysts based on an experience of Brazilian pathology service during 53 years.

BACKGROUND: Jaw pseudocysts are benign osseous lesions of unclear etiology. Among these, the simple bone cyst (SBC) and aneurysmal bone cyst (ABC) are intriguing bone pathologies still rarely studied together. This retrospective study aimed to present the long-term case series of patients with jaw pseudocysts focusing on the clinical, radiographic, and transoperative aspects. MATERIAL AND METHODS: A retrospective case series of patients with SBC and ABC was performed. Clinical, radiographic, and transoperative aspects of both pseudocysts were reviewed from the histopathological archives of 20,469 cases between 1959-2012. All descriptive data were summarized. RESULTS: Of 354 (15.25%) bone pathologies, 54 cases of jaw pseudocysts were found, with 42 (11.86%) SBC and 12 (3.39%) ABC cases. For both lesions, most of the sample were young Caucasian women with an asymptomatic posterior mandible lesion with undetermined time of evolution and none trauma history. A unique radiolucent scalloped lesion presenting an empty cavity were also observed for both conditions. However, some atypical findings were found for SBC including: the expansion of bone cortical, tooth resorption, displacement of the mandibular canal, and recurrence. The absence of painful symptoms and the lack of classical blood-filled cavity were observed in some cases of ABC. CONCLUSIONS: The SBC and ABC are bone pathologies with few retrospective studies, no previous studies on the two conditions, varied nomenclature, and atypical aspects in some cases. Therefore, the knowledge of clinical, imaging, and transoperative features of such pseudocysts are clinically valuable as diagnosis hypothesis of radiolucent lesions of the jaws.

Effects of bovine mammary gland biopsy and increased milking frequency on post-procedure udder health, histology, and milk yield.

Sixteen cows in early lactation were randomly distributed into two groups in order to evaluate the effects of mammary biopsies and increased milking frequency on tissue characteristics, post-biopsy udder health and histology. One group was milked twice a day (2×) starting on the 2nd day after calving, until 28 days in milk (DIM). The other group was milked four times a day (4×) from two to 21 DIM, and twice a day (2×) from 22 to 28 DIM. On days 2, 7, 14, 21, and 28 postpartum, one fragment of secretory tissue was collected from one mammary quarter at a time. Collections were alternated between the four mammary quarters per collection day. A total of 80 mammary tissue samples were collected. Qualitative and quantitative analyses of the tissues were conducted by histologic examination. Animal health was assessed by observation of feed intake behavior immediately after biopsy, and weight and body condition score before and one week after biopsy. Udder health was assessed daily from calving to 60 DIM with California Mastitis Test (CMT) and by noting alterations in the milk such as blood, milk clots, blood clots, clinical signs of mastitis. Milk composition and somatic cell count (SCC) were analyzed before and after the biopsies. Milk production was evaluated before biopsy, on the day of biopsy, and after the biopsy. An average of 10 fields at 40× magnification was obtained from each sample. There were no evident changes in mammary morphology as a result of milking two or four times/day at any of the evaluated time points. Biopsy wounds healed rapidly without infection. Intramammary bleeding and CMT alterations were observed in 96% and 75% of the biopsied mammary quarters, respectively. Clinical mastitis was diagnosed in 12% of the biopsied quarters. Different milking frequencies had no effect on the frequency and duration of post-biopsy alterations. Milk production decreased after biopsies done on days 2 for 2× and 4× groups, but it returned to pre-biopsy values within 1 day. Milk composition and SCC were affected transiently. Increased milking frequency did not influence udder health. Post-biopsy recovery was rapid and the procedure proved effective without damaging the cows' health.

Efeito de anti-inflamatórios não esteroidais na apoptose de células epidermais lamelares de equinos com laminite / Effect of nonsteroidal anti-inflammatory drugs on apoptosis of epidermal lamellar cells of equines with laminitis

O objetivo deste trabalho foi avaliar a influência da administração dos anti-inflamatórios não esteroidais (AINEs) cetoprofeno, fenilbutazona e flunixin meglumine sobre o índice apoptótico de células epiteliais do tecido lamelar de cavalos com laminite induzida por administração de amido. Foram empregados 20 equinos hígidos, divididos em quatro grupos experimentais (n=5): solução salina, cetoprofeno, fenilbutazona e flunixin meglumine. O tecido lamelar foi coletado por biópsia, fixado e corado pela técnica de TUNEL. À marcação positiva por essa técnica adicionou-se a avaliação morfológica celular para identificação da apoptose. Não houve diferença significativa no índice apoptótico entre os grupos tratados com anti-inflamatórios e o controle (P>0,05). Os anti-inflamatórios não interferiram sobre o índice apoptótico possivelmente porque foram administrados após a fase prodrômica da laminite e/ou porque não são eficazes em alterar a dinâmica da apoptose. Concluiu-se que a administração de anti-inflamatórios não esteroidais após a fase prodrômica da laminite não contribui para uma intervenção no curso da apoptose no tecido lamelar de cavalos com laminite quando comparados ao grupo controle não tratado. Outros estudos, com diferentes períodos de avaliação, são necessários para esclarecer os efeitos dos anti-inflamatórios não esteroidais na fisiopatologia da laminite em equinos, especialmente no que concerne à participação da apoptose.

The aim of this study was to evaluate the influence of the administration of anti-inflammatory drugs (NSAIDs) ketoprofen, phenylbutazone and flunixin meglumine on the apoptotic index of the epithelial cells of lamellar tissue of horses with induced laminitis. 20 healthy horses were employed and underwent induction of laminitis by administration of starch, divided into four groups with induced laminitis (n = 5): saline, ketoprofen, phenylbutazone and flunixin meglumine. The lamellar tissue was collected by biopsy, fixed and stained with the TUNEL technique. All that were stained positive by this technique were added to the cell morphological identification of apoptosis. No significant difference was found in the apoptotic index between the groups treated with anti-inflammatory and controls (P> 0.05). It was concluded that the administration of NSAIDs after the prodromal phase of laminitis does not contribute to an intervention in the course of apoptosis in the lamellar tissue of horses with laminitis when compared to the untreated control group. Other studies with different evaluation periods are needed to clarify the effects of anti-inflammatory non-steroidal drugs in the pathophysiology of laminitis in horses, especially regarding the role of apoptosis.

Efeito de anti-inflamatórios não esteroidais na apoptose de células epidermais lamelares de equinos com laminite / Effect of nonsteroidal anti-inflammatory drugs on apoptosis of epidermal lamellar cells of equines with laminitis

O objetivo deste trabalho foi avaliar a influência da administração dos anti-inflamatórios não esteroidais (AINEs) cetoprofeno, fenilbutazona e flunixin meglumine sobre o índice apoptótico de células epiteliais do tecido lamelar de cavalos com laminite induzida por administração de amido. Foram empregados 20 equinos hígidos, divididos em quatro grupos experimentais (n=5): solução salina, cetoprofeno, fenilbutazona e flunixin meglumine. O tecido lamelar foi coletado por biópsia, fixado e corado pela técnica de TUNEL. À marcação positiva por essa técnica adicionou-se a avaliação morfológica celular para identificação da apoptose. Não houve diferença significativa no índice apoptótico entre os grupos tratados com anti-inflamatórios e o controle (P>0,05). Os anti-inflamatórios não interferiram sobre o índice apoptótico possivelmente porque foram administrados após a fase prodrômica da laminite e/ou porque não são eficazes em alterar a dinâmica da apoptose. Concluiu-se que a administração de anti-inflamatórios não esteroidais após a fase prodrômica da laminite não contribui para uma intervenção no curso da apoptose no tecido lamelar de cavalos com laminite quando comparados ao grupo controle não tratado. Outros estudos, com diferentes períodos de avaliação, são necessários para esclarecer os efeitos dos anti-inflamatórios não esteroidais na fisiopatologia da laminite em equinos, especialmente no que concerne à participação da apoptose.(AU)

The aim of this study was to evaluate the influence of the administration of anti-inflammatory drugs (NSAIDs) ketoprofen, phenylbutazone and flunixin meglumine on the apoptotic index of the epithelial cells of lamellar tissue of horses with induced laminitis. 20 healthy horses were employed and underwent induction of laminitis by administration of starch, divided into four groups with induced laminitis (n = 5): saline, ketoprofen, phenylbutazone and flunixin meglumine. The lamellar tissue was collected by biopsy, fixed and stained with the TUNEL technique. All that were stained positive by this technique were added to the cell morphological identification of apoptosis. No significant difference was found in the apoptotic index between the groups treated with anti-inflammatory and controls (P> 0.05). It was concluded that the administration of NSAIDs after the prodromal phase of laminitis does not contribute to an intervention in the course of apoptosis in the lamellar tissue of horses with laminitis when compared to the untreated control group. Other studies with different evaluation periods are needed to clarify the effects of anti-inflammatory non-steroidal drugs in the pathophysiology of laminitis in horses, especially regarding the role of apoptosis.(AU)

Histopathological characterization of experimentally induced cutaneous loxoscelism in rabbits inoculated with Loxosceles similis venom

Envenomation by Loxosceles bites is characterized by dermonecrotic and/or systemic features that lead to several clinical signs and symptoms called loxoscelism. Dermonecrotic lesions are preceded by thrombosis of the dermal plexus. Recent studies show that atheromatous plaque is prone to thrombosis due to endothelial cell apoptosis. To the best of our knowledge, there are no reports of microscopic dermal lesion and endothelial cell apoptosis induced by Loxosceles similis venom in the literature. Thus, the aim of the present study is to describe histological lesions induced by L. similis venom in rabbit skin and to elucidate whether apoptosis of endothelial cells is involved in the pathogenesis of loxoscelism. Forty male rabbits were split into two groups: the control group (intradermally injected with 50 µL of PBS) and the experimental group (intradermally injected with 0.5 µg of L. similis crude venom diluted in 50 µL of PBS). After 2, 4, 6 and 8 hours of injection, skin fragments were collected and processed for paraffin or methacrylate embedding. Sections of 5 µm thick were stained by HE, PAS or submitted to TUNEL reaction. Microscopically, severe edema, diffuse heterophilic inflammatory infiltrate, perivascular heterophilic infiltrate, thrombosis, fibrinoid necrosis of arteriolar wall and cutaneous muscle necrosis were observed. Two hours after venom injection, endothelial cells with apoptosis morphology were evidenced in the dermal plexus. Apoptosis was confirmed by TUNEL reaction. It seems that endothelial cell apoptosis and its consequent desquamation is an important factor that induces thrombosis and culminates in dermonecrosis, which is characteristic of cutaneous loxoscelism.

Histopathological characterization of experimentally induced cutaneous loxoscelism in rabbits inoculated with Loxosceles similis venom

Envenomation by Loxosceles bites is characterized by dermonecrotic and/or systemic features that lead to several clinical signs and symptoms called loxoscelism. Dermonecrotic lesions are preceded by thrombosis of the dermal plexus. Recent studies show that atheromatous plaque is prone to thrombosis due to endothelial cell apoptosis. To the best of our knowledge, there are no reports of microscopic dermal lesion and endothelial cell apoptosis induced by Loxosceles similis venom in the literature. Thus, the aim of the present study is to describe histological lesions induced by L. similis venom in rabbit skin and to elucidate whether apoptosis of endothelial cells is involved in the pathogenesis of loxoscelism. Forty male rabbits were split into two groups: the control group (intradermally injected with 50 µL of PBS) and the experimental group (intradermally injected with 0.5 µg of L. similis crude venom diluted in 50 µL of PBS). After 2, 4, 6 and 8 hours of injection, skin fragments were collected and processed for paraffin or methacrylate embedding. Sections of 5 µm thick were stained by HE, PAS or submitted to TUNEL reaction. Microscopically, severe edema, diffuse heterophilic inflammatory infiltrate, perivascular heterophilic infiltrate, thrombosis, fibrinoid necrosis of arteriolar wall and cutaneous muscle necrosis were observed. Two hours after venom injection, endothelial cells with apoptosis morphology were evidenced in the dermal plexus. Apoptosis was confirmed by TUNEL reaction. It seems that endothelial cell apoptosis and its consequent desquamation is an important factor that induces thrombosis and culminates in dermonecrosis, which is characteristic of cutaneous loxoscelism.(AU)

Cell proliferation markers in the transplanted canine transmissible venereal tumor / Marcadores de proliferação celular no tumor venéreo transmissível canino transplantado

Adult male mongrel dogs were subcutaneously transplanted with the canine transmissible venereal tumor (TVT) on the hypogastric region. Twelve specimens of tumors were collected, half during the proliferative phase and the other half during the regressive phase. Fragments of the tumor were fixed in 10 percent buffered formalin and routinely processed for light microscopy. Sections of 4µm were stained by Schorr or AgNOR or either immunostained for MIB1 (Ki67). Schorr stain, AgNOR and MIB1 showed an increased proliferative activity through mitotic index, nuclear argyrophilic protein stain and cycling tumoral cells in the growing tumors, respectively. All of the three cell proliferation markers were able to distinguish the TVT in both evolution phases. MIB1 monoclonal antibody was the best in the morphologic evaluation of growth and regression of TVT. This resulted in higher values than AgNORs counting and mitotic index. MIB1 immunostaining was the most effective parameter of the proliferative activity of TVT. However, a significant correlation has been detected only between mitosis counting and AgNORs.

Cães machos, adultos, mestiços, foram transplantados com células do tumor venéreo transmissível canino (TVTC), na região hipogástrica. Foram coletados doze espécimes do TVTC, sendo metade durante a fase proliferativa e metade durante a fase regressiva. Fragmentos do tumor foram fixados em formol a 10 por cento, tamponado e processado rotineiramente para microscopia de luz. Secções de 4µm foram coradas pelo Shorr, ou pela AgNOR, ou ainda, imunocorado para MIB 1 (Ki67). As colorações pelo Shorr, AgNOR, ou MIB 1 mostraram um aumento do índice mitótico, coloração da proteína argirofílica nuclear e células tumorais ciclando em tumores em crescimento, respectivamente. Todos os três marcadores de proliferação celular foram capazes de distinguir o TVTC em ambas as fases de evolução. O anticorpo monoclonal MIB 1 foi o melhor na avaliação morfológica, de crescimento e regressão do TVTC. Isto resultou em um valor maior que a contagem de AgNOR e do índice mitótico. A imunomarcação com MIB1 foi o parâmetro mais efetivo da atividade proliferativa. No entanto, só foi observada uma correlação positiva entre a contagem de mitose e a AgNOR.

Marcadores de proliferação celular no tumor venéreo transmissível canino transplantado / Cell proliferation markers in the transplanted canine transmissible venereal tumor

Adult male mongrel dogs were subcutaneously transplanted with the canine transmissible venereal tumor (TVT) on the hypogastric region. Twelve specimens of tumors were collected, half during the proliferative phase and the other half during the regressive phase. Fragments of the tumor were fixed in 10 percent buffered formalin and routinely processed for light microscopy. Sections of 4µm were stained by Schorr or AgNOR or either immunostained for MIB1 (Ki67). Schorr stain, AgNOR and MIB1 showed an increased proliferative activity through mitotic index, nuclear argyrophilic protein stain and cycling tumoral cells in the growing tumors, respectively. All of the three cell proliferation markers were able to distinguish the TVT in both evolution phases. MIB1 monoclonal antibody was the best in the morphologic evaluation of growth and regression of TVT. This resulted in higher values than AgNORs counting and mitotic index. MIB1 immunostaining was the most effective parameter of the proliferative activity of TVT. However, a significant correlation has been detected only between mitosis counting and AgNORs.(AU)

Cães machos, adultos, mestiços, foram transplantados com células do tumor venéreo transmissível canino (TVTC), na região hipogástrica. Foram coletados doze espécimes do TVTC, sendo metade durante a fase proliferativa e metade durante a fase regressiva. Fragmentos do tumor foram fixados em formol a 10 por cento, tamponado e processado rotineiramente para microscopia de luz. Secções de 4µm foram coradas pelo Shorr, ou pela AgNOR, ou ainda, imunocorado para MIB 1 (Ki67). As colorações pelo Shorr, AgNOR, ou MIB 1 mostraram um aumento do índice mitótico, coloração da proteína argirofílica nuclear e células tumorais ciclando em tumores em crescimento, respectivamente. Todos os três marcadores de proliferação celular foram capazes de distinguir o TVTC em ambas as fases de evolução. O anticorpo monoclonal MIB 1 foi o melhor na avaliação morfológica, de crescimento e regressão do TVTC. Isto resultou em um valor maior que a contagem de AgNOR e do índice mitótico. A imunomarcação com MIB1 foi o parâmetro mais efetivo da atividade proliferativa. No entanto, só foi observada uma correlação positiva entre a contagem de mitose e a AgNOR.(AU)

Vero cells infected with the Lederle strain of canine distemper virus have increased Fas receptor signaling expression at 15 h post-infection.

We evaluated the expression of the Fas receptor gene in Vero cells infected with the Lederle vaccine strain of canine distemper virus using RT-PCR. Vero cells were plated, and after being grown for 24 h in MEM with 5% FBS, 80-90% confluent monolayer cultures were infected with the virus. The cells were harvested at 3, 6, 9, and 15 h post-infection. Uninfected Vero cells were used as a control. Total RNA was isolated from Vero cells using 1 mL Trizol(®) LS, and RT was performed using 2 µg total RNA. Primer pairs for RT-PCR amplification for the canine distemper virus nucleocapsid gene, the S26 reference gene, and the Vero rFas gene were used to analyze expression in Vero cells. RT-PCR results revealed virus activity at 3, 6, 9, and 15 h in the virus-infected Vero cells. The S26 housekeeping gene was amplified in virus infected and control samples. However, expression of the cell death receptor Fas was detected in Vero cells only at 15 h post-infection. We suggest that the Lederle vaccine induces apoptosis by Fas receptor signaling, possibly through caspase-8 signaling rather than through mitochondrial signaling in the infected cells.

Neurological disorder in cattle associated with bovine herpesvirus 4 / Desordem neurológica em bovinos associada ao herpevírus bovino 4

A nested PCR assay was used to diagnose bovine encephalitis through herpesviruses including bovine herpesvirus 5 (BHV-5), bovine herpesvirus 1 (BHV-1), Aujeszky's disease virus (SHV-1), and ovine herpesvirus 2 (OHV-2) in 14 fragments of central nervous system (CNS) from cattle that died with neurological signs. In addition, as some samples of bovine herpesvirus type 4 (BHV-4) have been isolated from neural tissue, it was also tested by nested PCR. The cases of encephalitis occurred in isolation at different times of the year and did not present any seasonality. The duration of the clinical course ranged between 1 to 15 days, and in 64.3 percent of the cases it manifested between 1 to 2 days. The most frequently observed neurological signs were ataxia, recumbency, unsteadiness and inability to stand, opisthotonus, paddling movements, nystagmus and ptyalism. In the nested assay, there was no evidence of: BHV-1, SHV-1 or OHV-2 in the DNA obtained from the CNS in any of the samples. But the presence of BHV-4 was found in all fragments of the CNS in cattle which died presenting neurological signs. Moreover, BHV-5 was found in association with BHV-4 in two of these samples.

Nested PCR foi utilizada para o diagnóstico de encefalite bovina por herpesvírus incluindo o herpesvírus bovino 5 (BHV-5), o herpesvírus bovino 1 (BHV-1), o vírus da doença de Aujeszky (SHV-1) e o herpesvírus ovino 2 (OHV-2) em 14 fragmentos do sistema nervoso central (SNC) de bovinos que morreram com sinais neurológicos. Embora o BHV-4 não seja reconhecido como vírus neurotrófico, foi detectado nos casos de encefalite que ocorreram isoladamente em diferentes épocas do ano e não apresentaram nenhuma sazonalidade. A duração do curso clínico variou entre 1 e 15 dias, e em 64,3 por cento dos casos manifestou-se entre 1 e 2 dias. Os sinais neurológicos mais freqüentemente observados foram ataxia, apatia, instabilidade, opistótono, movimentos de pedalagem, nistagmo e sialorréia. Nos ensaios de PCR nested realizados a partir do DNA obtido do SNC, não foi encontrado evidência de: BHV-1, SHV-1 ou OHV-2 em nenhuma das amostras. Mas, a presença de BHV-4 foi encontrada em todos os fragmentos do SNC de bovinos que morreram com sinais neurológicos. Além disso, o BHV-5 foi encontrado em associação com o BHV-4 em duas dessas amostras.

Neurological disorder in cattle associated with bovine herpesvirus 4 / Desordem neurológica em bovinos associada ao herpevírus bovino 4

A nested PCR assay was used to diagnose bovine encephalitis through herpesviruses including bovine herpesvirus 5 (BHV-5), bovine herpesvirus 1 (BHV-1), Aujeszky's disease virus (SHV-1), and ovine herpesvirus 2 (OHV-2) in 14 fragments of central nervous system (CNS) from cattle that died with neurological signs. In addition, as some samples of bovine herpesvirus type 4 (BHV-4) have been isolated from neural tissue, it was also tested by nested PCR. The cases of encephalitis occurred in isolation at different times of the year and did not present any seasonality. The duration of the clinical course ranged between 1 to 15 days, and in 64.3 percent of the cases it manifested between 1 to 2 days. The most frequently observed neurological signs were ataxia, recumbency, unsteadiness and inability to stand, opisthotonus, paddling movements, nystagmus and ptyalism. In the nested assay, there was no evidence of: BHV-1, SHV-1 or OHV-2 in the DNA obtained from the CNS in any of the samples. But the presence of BHV-4 was found in all fragments of the CNS in cattle which died presenting neurological signs. Moreover, BHV-5 was found in association with BHV-4 in two of these samples.(AU)

Nested PCR foi utilizada para o diagnóstico de encefalite bovina por herpesvírus incluindo o herpesvírus bovino 5 (BHV-5), o herpesvírus bovino 1 (BHV-1), o vírus da doença de Aujeszky (SHV-1) e o herpesvírus ovino 2 (OHV-2) em 14 fragmentos do sistema nervoso central (SNC) de bovinos que morreram com sinais neurológicos. Embora o BHV-4 não seja reconhecido como vírus neurotrófico, foi detectado nos casos de encefalite que ocorreram isoladamente em diferentes épocas do ano e não apresentaram nenhuma sazonalidade. A duração do curso clínico variou entre 1 e 15 dias, e em 64,3 por cento dos casos manifestou-se entre 1 e 2 dias. Os sinais neurológicos mais freqüentemente observados foram ataxia, apatia, instabilidade, opistótono, movimentos de pedalagem, nistagmo e sialorréia. Nos ensaios de PCR nested realizados a partir do DNA obtido do SNC, não foi encontrado evidência de: BHV-1, SHV-1 ou OHV-2 em nenhuma das amostras. Mas, a presença de BHV-4 foi encontrada em todos os fragmentos do SNC de bovinos que morreram com sinais neurológicos. Além disso, o BHV-5 foi encontrado em associação com o BHV-4 em duas dessas amostras.(AU)

Pancreatite crônica intersticial e caquexia causada por Eurytrema coelomaticum em vaca Nelore / Chronic interstitial pancreatitis and chronic wasting disease caused by Eurytrema coelomaticum in Nelore cow

A euritrematose bovina, causada por Eurytrema coelomaticum, tem sido relatada no Brasil. A doença pode afetar animais domésticos, incluindo ruminantes. Estes parasitos geralmente vivem nos ductos pancreáticos e ocasionalmente nos ductos biliares. Este estudo relata um quadro de caquexia em um bovino parasitado por E. coelomaticum proveniente de um rebanho de gado de corte localizado no Estado de Minas Gerais. O animal acometido apresentou perda progressiva de peso, glicosúria e cetonúria. Na necropsia o pâncreas apresentava-se diminuído de tamanho, brancacento, intensa e difusamente firme (fibrose). Vários ductos apresentavam-se dilatados e repletos de Eurytrema. Microscopicamente foi observada destruição extensa do parênquima pancreático e fibrose, ovos e parasitas intralesionais, hiperplasia ductal e inflamação multifocal crônica. Este relato descreve a perda progressiva de peso e pancreatite crônica associada a E. coelomaticum em bovino de corte no Estado de Minas Gerais.

Chronic interstitial pancreatitis and chronic wasting disease caused by Eurytrema coelomaticum in Nelore cow / Pancreatite crônica intersticial e caquexia causada por Eurytrema coelomaticum em vaca Nelore

A euritrematose bovina, causada por Eurytrema coelomaticum, tem sido relatada no Brasil. A doença pode afetar animais domésticos, incluindo ruminantes. Estes parasitos geralmente vivem nos ductos pancreáticos e ocasionalmente nos ductos biliares. Este estudo relata um quadro de caquexia em um bovino parasitado por E. coelomaticum proveniente de um rebanho de gado de corte localizado no Estado de Minas Gerais. O animal acometido apresentou perda progressiva de peso, glicosúria e cetonúria. Na necropsia o pâncreas apresentava-se diminuído de tamanho, brancacento, intensa e difusamente firme (fibrose). Vários ductos apresentavam-se dilatados e repletos de Eurytrema. Microscopicamente foi observada destruição extensa do parênquima pancreático e fibrose, ovos e parasitas intralesionais, hiperplasia ductal e inflamação multifocal crônica. Este relato descreve a perda progressiva de peso e pancreatite crônica associada a E. coelomaticum em bovino de corte no Estado de Minas Gerais.(AU)