Crotoxin stimulates an M1 activation profile in murine macrophages during Leishmania amazonensis infection.

American tegumentary leishmaniasis is caused by different species of Leishmania. This protozoan employs several mechanisms to subvert the microbicidal activity of macrophages and, given the limited efficacy of current therapies, the development of alternative treatments is essential. Animal venoms are known to exhibit a variety of pharmacological activities, including antiparasitic effects. Crotoxin (CTX) is the main component of Crotalus durissus terrificus venom, and it has several biological effects. Nevertheless, there is no report of CTX activity during macrophage - Leishmania interactions. Thus, the main objective of this study was to evaluate whether CTX has a role in macrophage M1 polarization during Leishmania infection murine macrophages, Leishmania amazonensis promastigotes and L. amazonensis-infected macrophages were challenged with CTX. MTT [3-(4,5dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide] toxicity assays were performed on murine macrophages, and no damage was observed in these cells. Promastigotes, however, were affected by treatment with CTX (IC50 = 22·86 µg mL-1) as were intracellular amastigotes. Macrophages treated with CTX also demonstrated increased reactive oxygen species production. After they were infected with Leishmania, macrophages exhibited an increase in nitric oxide production that converged into an M1 activation profile, as suggested by their elevated production of the cytokines interleukin-6 and tumour necrosis factor-α and changes in their morphology. CTX was able to reverse the L. amazonensis-mediated inhibition of macrophage immune responses and is capable of polarizing macrophages to the M1 profile, which is associated with a better prognosis for cutaneous leishmaniasis treatment.

Flavonoids from the Amazon plant Brosimum acutifolium induce C6 glioma cell line apoptosis by disrupting mitochondrial membrane potential and reducing AKT phosphorylation.

Glioblastoma, which is highly invasive and has a poor patient prognosis, is the most common type of brain tumor. Flavonoids have known antiproliferative and antineoplastic effects, such as apoptosis induction and tumor growth inhibition. We investigated the effects of treatment with three flavonoids (BAS-1, BAS-4, and BAS-6) isolated from the Amazon plant Brosimum acutifolium on the proliferation and migration of the C6 glioma cell line. Cytotoxicity was evaluated by MTT assay, and morphological changes were evaluated by phase-contrast microscopy and by transmission electron microscopy. Apoptosis was determined using Annexin V-FITC-propidium iodide (PI) staining. A hemolysis assay was used to evaluate plasma membrane injury. Antiproliferative effects were assessed by wound migration and colony formation assays. Mitochondrial transmembrane potential (ΔΨm) was determined using JC-1 dye and flow cytometry. To identify the flavonoid targets, western blotting was performed. BAS-1 and BAS-4 reduced C6 cell proliferation in a dose-dependent manner. BAS-6 showed no effect. Due to its high toxicity toward primary glial cells and its high hemolytic index, BAS-1 was not used in the remaining experiments. BAS-4 treatment did not induce cytotoxicity in primary glial cells; however, in glioma cells, it suppressed migration and invasion and led to apoptosis through mitochondrial damage, ΔΨm loss, cell cycle arrest, and reduced AKT phosphorylation, which is a component of the main cell survival pathway. We conclude that BAS-4 showed potential activity against glioma by inducing apoptosis mediated by ΔΨm loss and AKT pathway disruption, and future studies should further evaluate BAS-4 as a promising antineoplastic agent against glioblastoma.

Concomitant Malignant Pulmonary Peripheral Nerve Sheath Tumour and Benign Cutaneous Peripheral Nerve Sheath Tumour in a Dog.

Peripheral nerve sheath tumours (PNSTs) are neoplastic growths derived from Schwann cells, perineural cells or both. Malignant PNSTs (MPNSTs) are uncommon in domestic animals. This report describes the concomitant occurrence of PNSTs in a 10-year-old female cocker spaniel with a clinical history of respiratory impairment. Grossly, there was a large infiltrative mass in the caudal lobe of the right lung; smaller nodules were observed in the other lobes of the right lung. Furthermore, a small encapsulated cutaneous nodule was observed on the left hindlimb. Histopathology of the pulmonary tumours revealed the proliferation of pleomorphic spindle-shaped cells with moderate mitotic index arranged in interwoven bundles and concentric Antoni A and Antoni B patterns; invasion of the adjacent pulmonary tissue was observed. The cutaneous nodule consisted of neoplastic mesenchymal cells in interwoven bundles with concentric whorls, but without the marked anisokaryosis, binucleation and infiltrative growth seen in the pulmonary tumour. Immunohistochemistry revealed that both tumours were immunoreactive for vimentin, glial fibrillary acidic protein and S100 protein, but were negative for factor VIII. These findings are indicative of a MPNST in the lung with a concomitant benign PNST in the limb. This case represents the first report of a primary MPNST in the lung of a dog. This neoplastic growth should be included in the differential diagnosis of primary malignant pulmonary tumours of dogs.

Physalis angulata induces death of promastigotes and amastigotes of Leishmania (Leishmania) amazonensis via the generation of reactive oxygen species.

Leishmaniasis are a neglected group of emerging diseases that have been found in 98 countries and are caused by protozoa of the genus Leishmania. The therapy for leishmaniasis causes several side effects and leads to drug-resistant strains. Natural products from plants have exhibited activities against Leishmania in various experimental models. Physalis angulata is a widely used plant in popular medicine, and in the literature it has well-documented leishmanicidal activity. However, its mechanism of action is still unknown. Thus, this study aims to evaluate the mechanism driving the leishmanicidal activity of an aqueous extract of P. angulata root (AEPa). AEPa was effective against both promastigotes and intracellular amastigote forms of Leishmania amazonensis. This effect was mediated by an increase of reactive oxygen species (ROS), but not of nitric oxide (NO). The increased production of ROS induces cell death by phenotypes seems by apoptosis cell death in Leishmania, but not autophagy or necrosis. In addition, morphological analysis of macrophages showed that AEPa induced a high number of cytoplasmic projections, increased the volume of cytoplasm and number of vacuoles, caused cytoskeleton alterations and resulted in high spreading ability. AEPa also promoted superoxide anion (O2(-)) production in both uninfected macrophages and those infected with Leishmania. Therefore, these results revealed that AEPa causes cell death by phenotypes seems by apoptosis cell death in L. amazonensis and modulates macrophage activation through morphofunctional alterations and O2(-) generation to induce Leishmania death.

High mutation detection rate in TCOF1 among Treacher Collins syndrome patients reveals clustering of mutations and 16 novel pathogenic changes.

Twenty-eight families with a clinical diagnosis of Treacher Collins syndrome were screened for mutations in the 25 coding exons of TCOF1 and their adjacent splice junctions through SSCP and direct sequencing. Pathogenic mutations were detected in 26 patients, yielding the highest detection rate reported so far for this disease (93%) and bringing the number of known disease-causing mutations from 35 to 51. This is the first report to describe clustering of pathogenic mutations. Thirteen novel polymorphic alterations were characterized, confirming previous reports that TCOF1 has an unusually high rate of single-nucleotide polymorphisms (SNPs) within its coding region. We suggest a possible different mechanism leading to TCS or genetic heterogeneity for this condition, as we identified two families with no apparent pathogenic mutation in the gene. Furthermore, our data confirm the absence of genotype-phenotype correlation and reinforce that the apparent anticipation often observed in TCS families is due to ascertainment bias.

Callosal defect, microcephaly, severe mental retardation, and other anomalies in three sibs.

This study concerns an apparently lethal and previously undescribed syndrome of hypoplastic corpus callosum, microcephaly, severe mental retardation, preauricular skin tag, camptodactyly, growth retardation, and recurrent bronchopneumonia. This combination of anomalies is probably inherited as an autosomal recessive trait. A brief literature review of the anomalies or syndromes associated with callosal defects is discussed.

Waardenburg I syndrome: a clinical and genetic study of two large Brazilian kindreds, and literature review.

Two large kindreds with Waardenburg I syndrome are described. The total number of affected individuals is 73. The major manifestations are telecanthus (the only constant anomaly in all cases), prominent nasal root, round or square tip of nose, hypoplastic alae, smooth philtrum, bushy eyebrows with synophrys, sensorineural deafness, heterochromia or hypoisochromia iridis, hypopigmented ocular fundus, white forelock, premature greying, and hypopigmented skin lesions. These and other aspects of the syndrome, associated findings, frequency, genetic heterogeneity, pathogenesis, animal models, and gene linkage and mapping are reviewed briefly.

Multiple synostosis syndrome: study of a large Brazilian kindred.

We report a large Brazilian kindred with 28 cases of the autosomal dominant multiple synostosis syndrome. The main anomalies were symphalangism and carpal and tarsal synostoses. Other common findings included synostosis involving other bones, absence of phalanges and nails, short metacarpals, pes planovalgus with prominent lateral border, hypoplastic alae of nose, short upper lip, and dermatoglyphic abnormalities. This may be a variant of the WL syndrome.

A new case of the acromegaloid facial appearance syndrome?

An apparently new case of the acromegaloid facial appearance syndrome is reported. The main clinical findings were coarse facies and thickened lips, oral mucosa and upper eyelids. The patient also had macrocephaly with an arachnoid cyst in the right middle fossa, an anomaly not previously described in association with this syndrome.

[Ecologic aspects of American tegumentary leishmaniasis. 6. Anthropophillic Phlebotomus fauna of residual forests located in the Northeastern region of the State of São Paulo, Brazil]. / Aspectos ecológicos da leishmaniose tegumentar americana. 6. Fauna flebotomínea antropófila de matas residuais situadas na região centro-nordeste do Estado de São Paulo, Brasil.

This report shows the phlebotomine vectorial role in the endemic cutaneous leishmaniasis situated in the middle and north-eastern region of the São Paulo, Brazil. Analysis of information encloses a sandfly man-biting fauna of four different patches of residual forest. So, using the human-bait and Shannon trap for a period of one year we have caught 16,869 sandflies. The predominance of Lutzomyia intermedia (85.6%) was clear. Moreover, the low density of Lutzomyia whitmani and Lutzomyia pessoai and their epidemiological implications at present and in the past are discussed. Information about the daily activity of some species is given and we emphasize for the first time, the diurnal activity of Lutzomyia firmatoi in the São Paulo State.

[Triatoma infestans: importance, control and elimination of the species in the state of São Paulo, Brazil]. / Triatoma infestans: importância, controle e eliminação da espécie no Estado de São Paulo, Brasil.

Considerations about the control of the vectors of Chagas' disease are made in the State of Sao Paulo, mainly those activities that led to the elimination of T. infestans. First of all, the authors discuss different aspects of the biology of T. infestans mainly those that permitted it to adapt itself in rural areas of the State in the first middle of the century. Secondary factors that helped the control such as rural exodus are also analysed. The article shows that since 1965 the control became a campaign with different phases due to the epidemiological situation, the acquired knowledge and the entomological surveillance. After 25 years of work, the elimination of all the focus of T. infestans was finally reached and the campaign was ended. However, due to the possibility of reintroduction of the vector in rural areas by passive transportation besides the presence of secondary vectors (T. sordida and P. megistus) in several localities, the vector control activities were not interrupted and the surveillance is continuous.

[The prevalence, infection indices and feeding habits of the Triatominae captured in an area of epidemiological surveillance]. / Prevalência, índices de infecção e hábitos alimentares de triatomíneos capturados em uma área de vigilância epidemiológica.

Prevalence of triatomines and their infection by Trypanosoma cruzi, were studied during 1974-1981, in the Administrative Region of Campinas, São Paulo, Brazil. This investigation was continued (1982-1986), blood meal tests with bird, marsupial, rodent and human antisera having been added. Panstrongylus megistus, predominant in the region, whose winged instars are often found infected by T. cruzi in inhabited houses, displayed and alimentary eclecticism. From a sample of 7,785, 14.78 percent reacted positively with human antisera. A similar, but less marked behaviour was observed in Rhodnius neglectus. Triatoma sordida, the second most important species in the Region, was neither demonstrated to be feeding on humans nor to be infected by T. cruzi. Changes in their natural habitat probably caused sharp rises in T. arthurneivai populations in the human dwellings. The capture of three specimens of Microtriatoma borbai marks the first occurrence of this species in the State São Paulo. "Focus investigation" in areas in the surveillance phase can detect triatomines in the process of domiciliation.

[Triatoma brasiliensis Neiva, 1911 (Hemiptera, Reduviidae): observations on dynamics of feeding behavior in an artificial ecotope]. / Triatoma brasiliensis (Neiva,1911) (Hemiptera, Reduviidae): observações sobre seu comportamento em relação fonte alimentar em galinheiro experimental.

In order to study the behavior of triatomine bugs under artificial ecotopic conditions, an experimental, double-walled poultry coop was built. Its inside walls were made of sun-dried clay blocks which could be broken apart for better access to the insects hiding in their crevices. The coop was subdivided into two main areas of different sizes using wire netting and a wall made of perforated bricks. The chickens, the only food source available for the triatomines, were kept in the larger, rectangular area, further subdivided diagonally. The triatomines were released in the smaller area and were allowed to migrate to the larger one though the holes in the bricks. Information about migration, survival, and hiding places of fourth-stage nymphs of Triatoma brasiliensis was thus gathered. At 30-day intervals, over a period of six months, the inner walls of the house were taken apart and inspected. The nymphs were marked with different colors according to the month and place of capture. It was found that 88.5% of the nymphs were captured near the roost and found engorged. After having taken their first blood meal, they would remain in the vicinity of their food source. The results obtained are considered epidemiologically relevant, and particularly so as pertaining to the control of the vectors of Chagas disease.

[Causes of pre-verbal deafness in an institutionalized population, emphasizing genetic etiology]. / Causas de surdez pré-verbal em uma população institucionalizada, enfatizando a etiologia genética.

OBJECTIVE: To investigate the causes of pre-verbal deafness in an institutionalized population emphasizing genetic etiology, considering the scarceness of national data in this field. METHODS: Based on the promptuaries of 658 pupils from 18 institutions for deaf people in Recife, information about audiological tests, laboratory examinations, etiology, associated anomalies, consanguinity and other deaf persons in the family was obtained; dysmorphologic examination was performed in 557 pupils; all the families with recurrent cases of deafness were investigated, having the pertinent pedigrees been constituted. RESULTS: About 13% of the studied population probably manifest genetic deafness; the autosomal recessive, autosomal dominant and X-linked recessive patterns of inheritance were observed, respectively, in 87.7%, 8.8% and 3.5% of the familial cases; among the examined pupils, 4.3% presented specific dysmorphic syndromes or anomalies associated with deafness; the acquired causes and unknown etiology were represented by 41.5% and 45.5% respectively. CONCLUSIONS: Obtained frequencies for the distinct causal factors of pre-verbal deafness are within the spectrum of variation of the frequencies observed in similar works, pointing out that in a great number of cases the etiology is unknown.

[Genetic deafness: a brief review]. / Surdez genética: uma revisão sucinta.

In this article, we present a brief literature review of hereditary deafness, considering the main historical aspects, genetic heterogeneity, gene mapping and the problems related to genetic counseling.

[Cockayne syndrome in two brothers]. / Síndrome de Cockayne em dois irmãos.

We report the clinical history of two brothers with the classical Cockayne syndrome. The main manifestations consisted of cachectic dwarfism, mental retardation, intracranial calcifications, microcephaly, enophthalmos, senile appearance, joint hypomotility and skin photosensitivity. In one of these children, who died at 10 years of age of bronchopneumonia, necropsy studies revealed a variety of anomalies, mainly encephalic,which included an arachnoidal cyst at the base of the cerebellum, a defect apparently previously undescribed inpatients with this syndrome.

Accidentes ofídicos notificados al Programa Nacional de Control de Zoonosis y Centro Antirrábico Nacional, Paraguay (2015) / Ophidic related accidents notified to the National Program of Control of Zoonosis and National Anti-rabies Center, Paraguay (2015)

Introducción: Las picaduras de serpientes constituyen un grave problema de Salud Pública. Objetivo: Describir los casos de mordeduras de serpientes notificados al Programa Nacional de Control de Zoonosis y Centro Antirrábico Nacional, Paraguay durante el año 2015. Método: Estudio observacional, descriptivo, retrospectivo y de corte transversal. Los criterios de inclusión fueron toda persona que haya sido mordida por alguna serpiente y que haya sido notificado al Programa Nacional de Control de Zoonosis y Centro Antirrábico Nacional durante el año 2015. Tipo de muestreo no probabilístico. Resultados: Fueron notificados 169 casos de mordeduras de serpientes. De acuerdo con las características sociodemográficas, la mayoría de los afectados fueron varones, con una edad promedio de 26,39 ±1.36. Los lugares más frecuentes donde ocurrieron los accidentes ofídicos fueron las chacras, y en el Departamento de San Pedro. En la mayoría de las mordeduras no se pudo identificar a la serpiente agresora, y cuando fueron identificadas, fueron las del genero Bothrops, las más frecuentes. En cuanto a la localización de las picaduras, fueron más frecuentes en los miembros inferiores, siendo necesario la hospitalización para el tratamiento. No se reportaron fallecidos. Conclusión: En el año 2015 fueron notificados 169 casos, los departamentos con mayor prevalência fueron San Pedro, Itapúa y Caazapá. El perfil del afectado es un varón con promedio de edad de 26 años, trabajador rural. Las serpientes del género Bothrops causaron el 32,5% de las mordeduras. Palabras clave: Mordeduras de serpiente; Envenenamiento; Epidemiología.

Introduction: Snake bites are a serious public health problem. Objective: To describe the cases of snake bites reported to the National Zoonosis Control Program and National Anti-Rabies Center, Paraguay during 2015. Method: Observational, descriptive, retrospective and cross-sectional study. The inclusion criteria were any person who has been bitten by a snake and it has been notified to the National Zoonosis Control Program and National Anti-Rabies Center during 2015. Type of non-probabilistic sampling. Results: 169 cases of snake bites were reported. According to the sociodemographic characteristics, the majority of those affected were males, with an average age of 26.39 ± 1.36. The most frequent places where theophidic accidents occurred were the farms, and in the Department of San Pedro. In most of the bites the aggressor snake could not be identified, and when they were identified, they were those of the genus Bothrops, the most frequent. As for the location of the bites, they were more frequent in the lower limbs, requiring hospitalization for treatment. No deaths were reported. Conclusion: In the year 2015, 169 cases were notified, the departments with the highest prevalence were San Pedro, Itapua and Caazapá. The profile of the affected is a male with an average age of 26 years, a rural worker. The snakes of the genus Bothrops caused 32.5% of the bites. Keywords: Snake bites; Poisoning; Epidemiology.

Helicobacter spp. infection induces changes in epithelial proliferation and E-cadherin expression in the gastric mucosa of pigs.

Gastric disease is common in finishing pigs. Helicobacter spp. infection has been associated with gastritis, gastric ulcers and gastric neoplasia in man and animals. The aim of this study was to determine the effects of Helicobacter spp. infection on gastric morphology in pigs, with emphasis on glandular cell proliferation and E-cadherin expression. Samples of fundus and antrum from 67 finishing pigs were examined microscopically and by immunohistochemistry. The presence of Helicobacter spp. was confirmed by polymerase chain reaction (PCR). Mucosal changes were evaluated and epithelial proliferation was determined by evaluation of the morphometry of nucleolar organizer regions and counting proliferating cell nuclear antigen-positive cells and mitotic figures. Intercellular adhesion was evaluated by E-cadherin expression. In 47 (70%) pigs, Helicobacter spp. infection was confirmed by PCR. Histological findings associated with the infection included mononuclear cell infiltration of the lamina propria and glandular degeneration. There was a significant association between infection and epithelial proliferation in both regions as well as a decrease in the expression of E-cadherin in the antrum.

Tumor maligno da bainha perineural retrobulbar e intracraniano em cão: relato de caso / Malignant neoplasm of the perineural sheath retrobulbar and intracranial in dog: case report

Os tumores da bainha perineural são pouco frequentes em animais domésticos. Relata-se o caso de um cão, sem raça definida, de cinco anos, fêmea, com histórico de aumento de volume em região retrobulbar do globo ocular direito (GOD) havia três semanas. Ao exame clínico, constatou-se a presença de uma neoformação retrobulbar direita com deslocamento do globo ocular cranialmente. Aos exames realizados durante o internamento, não foi possível localizar a origem e a extensão da neoformação. O animal veio a óbito por parada cardiorrespiratória e foi encaminhado para a necropsia. À avaliação macroscópica, observou-se neoformação esbranquiçada fixada à base do crânio em região selar, com possível origem no terceiro (III) par de nervos cranianos, a qual se infiltrava no encéfalo na altura do hipotálamo, estendia-se caudalmente em direção ao tronco encefálico e cranialmente à órbita direita, comprimindo, assim, o GOD. Microscopicamente consistia de feixes curtos entrelaçados ou enovelados de células fusiformes com pleomorfismo discreto a moderado, alternando-se a áreas de necrose multifocalmente, compatível com tumor da bainha perineural. Ao exame imuno-histoquímico, apresentou marcação fraca para S100 e GFAP e marcação positiva para vimentina, o que indica caráter maligno.(AU)

Tumors of the perineural sheath are uncommon in domestic animals. We report the case of a 5-year-old female dog with a history of increased volume in the retrobulbar region of the right eye (RE) three weeks ago. The clinical examination revealed a presence of a right retrobulbar neoformation with cranial ocular globe displacement. In the examinations carried out during hospitalization, it was not possible to locate a source and an extension of the neoformation. The animal died of cardiorespiratory arrest and was referred to an autopsy. The macroscopic evaluation revealed a whitish neoformation fixated to the base of the skull in a seal region, with a possible non-III origin of cranial nerves, infiltrating non-encephalon at the height of the hypothalamus, extending caudally towards the brainstem and cranially to the orbit right, compressing RE. Microscopically it consisted of short bundles intertwined or enovelados of spindle cells with discrete to moderate pleomorphism, alternating to areas of multifocal necrosis, compatible with tumor of the perineural sheath. Immunohistochemical examination showed weak marking for S100 and GFAP and positive marking for vimentin, indicating malignancy.(AU)

Malignant pilomatricoma in a dog.

An 11-year-old female German shepherd dog was presented with a history of lameness and pain in the left forelimb. Clinical examination revealed ataxia of the hindlimbs and a subcutaneous mass in the left prescapular region. Radiography revealed metastatic foci in the left humerus, lung and abdomen. Gross necropsy examination revealed a firm, white mass in the left prescapular region. Multiple nodules with similar characteristics were observed in the lung, liver and spleen. Bone lysis was noted in the humerus and the fifth to seventh lumbar vertebrae. Microscopical examination revealed a proliferation of basal cells forming irregular islands of various sizes and surrounding extensive zones of keratinized 'ghost' cells. A definitive diagnosis of malignant pilomatricoma was made. This is a rare tumour in dogs with no previous report of metastasis to the spleen and liver.