Progressive Purkinje Cell Degeneration in tambaleante Mutant Mice Is a Consequence of a Missense Mutation in HERC1 E3 Ubiquitin Ligase

The HERC gene family encodes proteins with two characteristic domains: HECT and RCC1-like. Proteins with HECT domain shave been described to function as ubiquitin ligases, and those that contain RCC1-like domains have been reported to function as GTPases regulators. These two activities are essential in a number of important cellular processes such as cell cycle, cell signaling, and membrane trafficking. Mutations affecting these domains have been found associated with retinitis pigmentosa, amyotrophic lateral sclerosis, and cancer. In humans, six HERC genes have been reported which encode two subgroups of HERC proteins: large (HERC1-2) and small (HERC3-6). The giant HERC1 protein was the first to be identified. It has been involved in membrane trafficking and cell proliferation/growth through its interactions with clathrin, M2-pyruvate kinase, and TSC2 proteins. Mutations affecting other members of the HERC family have been found to be associated with sterility and growth retardation. Here, we report the characterization of a recessive mutation named tambaleante, which causes progressive Purkinje cell degeneration leading to severe ataxia with reduced growth and lifespan in homozygous mice aged over two months. We mapped this mutation in mouse chromosome 9 and then performed positional cloning. We found a GuA transition at position 1448, causing a Gly to Glu substitution (Gly483Glu) in the highly conserved N- terminal RCC1-like domain of the HERC1 protein. Successful transgenic rescue, with either a mouse BAC containing the normal copy of Herc1 or with the human HERC1 cDNA, validated our findings. Histological and biochemical studies revealed extensive autophagy associated with an increase of the mutant protein level and a decrease of mTOR activity. Our observations concerning this first mutation in the Herc1 gene contribute to the functional annotation of the encoded E3 ubiquitin ligase and underline the crucial and unexpected role of this protein in Purkinje cell physiology.

Hemoglobina extracelular gigante de Glossoscolex paulistus: um extraordinário sistema supramolecular hemoproteico

Giant extracellular hemoglobins are considered the summit of complexity in systems that carry oxygen, constituting an extraordinary model system to the study of hemoproteins. This class includes the hemoglobin of the annelid Glossoscolex paulistus that presents high cooperativity, great oligomeric and redox stabilities and ability of oligomeric reassociation. These properties have motivated evaluations about its utilization as prototype of artificial blood and biosensor. Kinetic studies involving autoxidation and detailed spectroscopic characterizations of its ferrous and ferric species have propitiated information about the structure-activity relationship of this macromolecule. The present review analyzes several biochemical issues, evaluating the state-of-art of this subject.

Reencontro de Balanoglossus gigas Fr. Muller no litoral brasileiro

Re-finding of Balanoglossus gigas FR. MUULLER on the brasilian sea shore. Balanoglossus gigas, the giant Enteropneusta, has been described for the first time by Fritz Muller in his notes collected and published by Dr. HERMANN VON IHERING (1898, pg. 35). FR. MULLER found the Balanoglossus on the coats of the State of Santa Catarina in 1884. Once again in 1885 several specimens of that animal were captured by FR. MULLER in the same place. Since that time up to now no references on the occurrence of this animal have been found in the zoological bibliography. During the spring of 1948 Prof. W. BESNARD, Director of he Instituto Paulista de Oceanografia saw some signals indicating the existence of Balanoglossus at the beach of São Sebastião, State of São Paulo. From 1948 to now several attempts have been made to catch the animal alive and complete. On the last September during a shorts visit to the beach of São Sebastião one Balanoglossus was captured and brought to the Department of General and Animal Physiology in good conditions. The animal measured 1.80 m in length. It seems to be the largest Balanoglossus known. According to the descriptions of SPENGEL (1893, pg. 158), and VAN DER HORST (1939) pg. 717) it was possible to identify this Enteropneusta as Balanoglossus gigas FR. MULLER, refound at the Brazilian coast sixty six years after its discovery by FR. MÜLLER.

Mössbauer spectra of iron oxides in the bulk and nanocrystalline states

Magnetic nanoparticles are very important in modern industry. These particles are used in many different spheres of life. Nanoparticles have unusual physical and chemical properties connected both with quantum dimensional effects and with the increased role of the surface atoms. Most clearly the difference between the properties of bulk materials and nanoparticles can be seen in the magnetic properties of these materials. The most typical magnetic properties of nanomaterials are superparamagnetism with the size of the cluster from 1 to 10 nm; single-domain magnetic state of nanoclusters and nanostructures up to 20 nm; magnetization processes connected with magnetic cluster ordering and with its forms and sizes; quantum magnetic tunneling effects when magnetization changes by jumps and giant magnetoresistance effects. For research of the magnetic properties of iron-containing nanostructures, it is convenient to apply Mӧssbauer spectroscopy. In this work a number of nano-sized samples of iron oxides were examined by Mössbauer spectroscopy. The Mössbauer spectra of nanoparticles with various sizes were obtained. Mössbauer spectra of iron oxide nanoparticles were compared with the spectra of bulk samples. It was shown how the spectra of iron oxide nanoparticles change depending on the particle sizes.

Growth, magnetic and transport properties of InSb and II-IV-As2 semiconductors doped with manganese

This thesis is devoted to growth and investigations of Mn-doped InSb and II-IV-As2 semiconductors, including Cd1-xZnxGeAs2:Mn, ZnSiAs2:Mn bulk crystals, ZnSiAs2:Mn/Si heterostructures. Bulk crystals were grown by direct melting of starting components followed by fast cooling. Mn-doped ZnSiAs2/Si heterostructures were grown by vacuum-thermal deposition of ZnAs2 and Mn layers on Si substrates followed by annealing. The compositional and structural properties of samples were investigated by different methods. The samples consist of micro- and nano- sizes clusters of an additional ferromagnetic Mn-X phases (X = Sb or As). Influence of magnetic precipitations on magnetic and electrical properties of the investigated materials was examined. With relatively high Mn concentration the main contribution to magnetization of samples is by MnSb or MnAs clusters. These clusters are responsible for high temperature behavior of magnetization and relatively high Curie temperature: up to 350 K for Mn-doped II-IV-As2 and about 600 K for InMnSb. The low-field magnetic properties of Mn-doped II-IV-As2 semiconductors and ZnSiAs2:Mn/Si heterostructures are connected to the nanosize MnAs particles. Also influence of nanosized MnSb clusters on low-field magnetic properties of InMnSb have been observed. The contribution of paramagnetic phase to magnetization rises at low temperatures or in samples with low Mn concentration. Source of this contribution is not only isolated Mn ions, but also small complexes, mainly dimmers and trimmers formed by Mn ions, substituting cation positions in crystal lattice. Resistivity, magnetoresistance and Hall resistivity properties in bulk Mn-doped II-IV-As2 and InSb crystals was analyzed. The interaction between delocalized holes and 3d shells of the Mn ions together with giant Zeeman splitting near the cluster interface are respond for negative magnetoresistance. Additionally to high temperature critical pointthe low-temperature ferromagnetic transition was observed Anomalous Hall effect was observed in Mn doped samples and analyzed for InMnSb. It was found that MnX clusters influence significantly on magnetic scattering of carriers.

Pseudo-aneurisma gigante de artéria axilar esquerda

A case of a gunshot of the left axilary artery with a giant pseudoaneurism formation with sixteen years of evolution is presented. The 61- years-old male patient present since them a small assintomatic tumoration in infraclavicular localization. In the last eight months the tumor became symptomatic and volumous, reaching 20 cm in diameter, with signals of infection and imminent rupture. The patient was operated on with ligature of the axilary artery and evacuation of the pseudoaneurysm. The surgical handling and endovascular therapeutic options available in the present time are discussed. A short review about the disease as well as a bibliographic update are present. The authors also call attention for the necessity of a correct management of the vascular injures in the event of the first management.

Perfuração gástrica secundária a tricobezoar gigante: descrição de caso e revisão da literatura

Gastric bezoars are impactations offoreign material in lhe stomach. When they are caused by hail; they are named tricho- bezoars. The complications oftrichobezoars are very rare. In this papel; we describe a case of a 16-year-old girl that had a previous history oftricophagia, and had an acute abdominal pain with a pneumoperitoneum in the abdomen radiography. An operation was performed and a gastric perforation was founded associated with a giant trichobezoa7: The trichobezoar was removed by traction through a gastrostomy which was performed in order to remove lhe trichobezoa7: Some fragments of the ulcer were obtained to histological study. The gastrostomy was treated by a gastrorraphy confection. In lhe post- operative period a left subfrenic abscess was revealed and has been drained by laparatomy 15 days after the fisrt operation. After the second surgical procedure the patient had a good evolution, and left the hospital in good health conditions.

Adenoma viloso com transformação carcinomatosa da ampola de Vater

Villous adenomas of the duodenum and ampulla of Vater are uncommon, but they have been diagnosed more frequently with the increasing use of upper gastrointestinal endoscopy. Differential diagnosis with villous adenocarcinoma may be difficult. The authors present a case of a 47-year-old man with a giant villous adenoma of the duodenum, with intermittent jaundice, that was treated by pancreatoduodenectomy.

Hepaticojejunostomia em "y" de roux videoassistida

Management of common bile duct stones in the era of laparoscopic surgery is controversial. The biliary anastomosis is indicated in case of large common bile duct, recurrent stones, giant stones and concomitant common bile duct stricture and duct stones. The development of laparoscopic techniques permits to perform this type of surgery laparoscopically as well as the open procedure.

Generation and Characterization of Knockout Mice and Analysis of Patient Material Demonstrates a Role for Tissue Inhibitor of Metalloproteinases 4 (Timp4) in the Cardiovascular System and Tumor Metastasis

This thesis focuses on tissue inhibitor of metalloproteinases 4 (TIMP4) which is the newest member of a small gene and protein family of four closely related endogenous inhibitors of extracellular matrix (ECM) degrading enzymes. Existing data on TIMP4 suggested that it exhibits a more restricted expression pattern than the other TIMPs with high expression levels in heart, brain, ovary and skeletal muscle. These observations and the fact that the ECM is of special importance to provide the cardiovascular system with structural strength combined with elasticity and distensibility, prompted the present molecular biologic investigation on TIMP4. In the first part of the study the murine Timp4 gene was cloned and characterized in detail. The structure of murine Timp4 genomic locus resembles that in other species and of the other Timps. The highest Timp4 expression was detected in heart, ovary and brain. As the expression pattern of Timp4 gives only limited information about its role in physiology and pathology, Timp4 knockout mice were generated next. The analysis of Timp4 knockout mice revealed that Timp4 deficiency has no obvious effect on the development, growth or fertility of mice. Therefore, Timp4 deficient mice were challenged using available cardiovascular models, i.e. experimental cardiac pressure overload and myocardial infarction. In the former model, Timp4 deficiency was found to be compensated by Timp2 overexpression, whereas in the myocardial infarct model, Timp4 deficiency resulted in increased mortality due to increased susceptibility for cardiac rupture. In the wound healing model, Timp4 deficiency was shown to result in transient retardation of re-epithelialization of cutaneous wounds. Melanoma tumor growth was similar in Timp4 deficient and control mice. Despite of this, lung metastasis of melanoma cells was significantly increased in Timp4 null mice. In an attempt to translate the current findings to patient material, TIMP4 expression was studied in human specimens representing different inflammatory cardiovascular pathologies, i.e. giant cell arteritis, atherosclerotic coronary arteries and heart allografts exhibiting signs of chronic rejection. The results showed that cardiovascular expression of TIMP4 is elevated particularly in areas exhibiting inflammation. The results of the present studies suggest that TIMP4 has a special role in the regulation of tissue repair processes in the heart, and also in healing wounds and metastases. Furthermore, evidence is provided suggesting the usefulness of TIMP4 as a novel systemic marker for vascular inflammation.

Doença de castleman simulando neoplasia pancreática

A 33-year-old man presented to our department with a 4-month history of right quadrant abdominal pain. Physical examination was normal. A chest X-ray showed no remarkable findings. Ultrasonography demonstrated a hypoechoic mass measuring 6 cm in the head of the pancreas. Computed tomography confirmed a solid mass in the pancreas without Wirsung or bile duct dilatation. At laparotomy, excision a 6 cm egg-shaped and hypervascular mass in the head of the pancreas was performed. Histologically, the features were consistent with Castleman disease. Castleman's disease is a rare, usually benign lymphoid condition described by Castleman (1954) and characterized by giant lymph nodes. Surgical resection is diagnostic and curative.

Pseudoaneurisma gigante de artéria esplênica

Splenic artery pseudoaneurysm larger than 10 cm is a rare condition. The risk of rupture is probably high and surgical treatment is necessary. The objective of this article is to report a case of a patient with giant pseudoaneurysm of the splenic artery submitted to surgical resection. A 26-year-old man complaining of gastrointestinal hemorrhage and abdominal pain The patient’s medical history revealed that one year before he had an abdominal blunt trauma. The angiography showed a giant pseudoaneurysm of the splenic artery with compression of the stomach. The patient was operated on by abdominal access and the spleen and pseudoaneurysm were resected. The postoperative course was uneventful and the patient was discharged 13 days after surgery without problems.

Baço migratório em hérnia paraestomal

The authors report a case of a parastomal hernia that incarcerated a wandering spleen. We present a very rare case of an 85-year-old woman with a giant parastomal hernia. Computed tomography revealed a parastomal hernia with a wandering spleen inside. We introduce a different and simple alternative approach, with the smallest inconvenience as possible to solve the trouble. We carried out the treatment with a prosthetic repair closing the defect with ePTFE mesh, performed through a laparoscopic approach, intraperitoneal, like a tie and move the spleen back to its anatomical place. The result was excellent.

A retrospective search for bovine respiratory syncytial virus (BRSV) antigens in histological specimens by immunofluorescence and immunohistochemistry

Bovine respiratory syncytial virus (BRSV) has been only sporadically identified as a causative agent of respiratory disease in Brazil. This contrasts with frequent reports of clinical and histopathological findings suggestive of BRSV-associated disease. In order to examine a possible involvement of BRSV in cases of calf pneumonia, a retrospective search was performed for BRSV antigens in histological specimens submitted to veterinary diagnostic services from the states of Rio Grande do Sul and Minas Gerais. Ten out of 41 cases examined (24.4%) were positive for BRSV antigens by immunohistochemistry (IPX). Eight of these cases (19.5%) were also positive by indirect immunofluorescence (IFA), and 31 cases (75.6%) were negative in both assays. In the lungs, BRSV antigens were predominantly observed in epithelial cells of bronchioles and less frequently found in alveoli. In one case, antigens were detected only in the epithelium of the alveolar septae. The presence of antigen-positive cells was largely restricted to epithelial cells of these airways. In two cases, positive staining was also observed in cells and cellular debris in the exudate within the pulmonary airways. The clinical cases positive for BRSV antigens were observed mainly in young animals (2 to 12 month-old) from dairy herds. The main microscopic changes included bronchointerstitial pneumonia characterized by thickening of alveolar septae adjacent to airways by mononuclear cell infiltrates, and the presence of alveolar syncytial giant cells. In summary, the results demonstrate the suitability of the immunodetection of viral antigens in routinely fixed tissue specimens as a diagnostic tool for BRSV infection. Moreover, the findings provide further evidence of the importance of BRSV as a respiratory pathogen of young cattle in southeastern and southern Brazil.

The materno-fetal interface in llama (Lama guanicoe glama)

Samples from 9 llamas (28 through 36 weeks of gestation) were collected and fixed in 4% buffered paraformaldehyde (light microscopy) and in 2.5% buffered glutaraldehyde (transmission and scanning electron microscopy). The material was processed in paraplast and slides (5mm) were stained with HE, PAS, Masson-Trichrome, acid phosphatase and Perl's. The uteroferrin was immunolocalized. The results show that llama placenta is chorioallantoic, diffuse, folded and epitheliochorial, and the fetus is covered with an epidermal membrane. The trophoblast cells have variable morphology: cubic, rounded and triangular cells, with cytoplasm containing PAS-positive granules. Binucleated cells with large cytoplasm and rounded nuclei, as well as giant trophoblastic cells with multiple nuclei were also observed. Numerous blood vessels were observed beneath the cells of the uterine epithelium and around the chorionic subdivided branches. Glandular activity was shown by PAS, Perl's, and acid phosphatase positive reactions in the cytoplasm and glandular lumen, and by immunolocalization of the uteroferrin in the glandular epithelium. The uterine glands open in spaces formed by the areoles, which are filled by PAS-positive material. The llama fetus was covered by the epidermal membrane, composed of stratified epithelium, with up to seven layers of mono-, bi- or trinucleated cells. The high level of maternal and fetal vascularization surfaces indicates an intense exchange of substances across both surfaces. The metabolic activity shown in the uterine glands suggests an adaptation of the gestation to the high altitudes of the natural habitat of this species.