Expression of cyclooxygenase enzymes and prostaglandin E2 receptors in inflammatory airway diseases

Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina

Natural infection of phlebotomines (Diptera: Psychodidae) in a visceral-leishmaniasis focus in Mato Grosso do Sul, Brazil

The main purpose of this study was to investigate natural infection by Leishmania in phlebotomine females in a visceral-leishmaniasis focus in Antonio João county in Mato Grosso do Sul State, Brazil. Between June and October 2003, the digestive tracts of 81 females captured in Aldeia Campestre, Aldeia Marangatu and Povoado Campestre were dissected. The females were separated by species, location, area and date of capture into 13 groups and kept in ethanol 70%. To identify the Leishmania species using the PCR technique, amplifications of the ribosomal-DNA (rDNA) and mini-exon genes were analyzed. Of the 81 specimens, 77 (95%) were Lutzomyia longipalpis, making this the most common species; only one specimen of each of the species Brumptomyia avellari, Evandromyia cortelezzii, Evandromyia lenti and Nyssomyia whitmani was found. Trypanosomatids were identified in eight of the nine groups of Lutzomyia longipalpis (10.39%) one group from Aldeia Campestre, one from Aldeia Marangatu and six from Povoado Campestre; of the eight groups, one from Aldeia Marangatu and another, with promastigotes forms also confirmed by dissection (1.23%) from Povoado Campestre, were identified by PCR as Leishmania chagasi (2.6%). The other groups gave negative results. These findings indicate that there is a high risk of leishmaniasis transmission in this area.

Spatial distribution of dengue in the city of Cruzeiro, São Paulo State, Brazil: use of geoprocessing tools

The aim of this article is to identify patterns in spatial distribution of cases of dengue fever that occurred in the municipality of Cruzeiro, State of São Paulo, in 2006. This is an ecological and exploratory study using the tools of spatial analysis in the preparation of thematic maps with data from Sinan-Net. An analysis was made by area, taking as unit the IBGE census, the analysis included four months in 2006 which show the occurrence of the disease in the city. The thematic maps were constructed by TerraView 3.3.1 software, the same software provided the values of the indicators of Global Moran (I M) every month and the Kernel estimation. In the year 2006, 691 cases of dengue were georeferenced (with a rate of 864.2 cases/100,000 inhabitants); the indicators of Moran and p-values obtained were I M = 0.080 (March) p = 0.11; I M = 0.285 (April) p = 0.01; I M = 0.201 (May) p = 0.01 and I M = 0.002 (June) p = 0.57. The first cases were identified in the Northeast and Central areas of Cruzeiro and the recent cases, in the North, Northeast and Central. It was possible to identify census tracts where the epidemic began and how it occurred temporally and spatially in the city.

LIVER BIOPSY: IMPORTANCE OF SPECIMEN SIZE IN THE DIAGNOSIS AND STAGING OF CHRONIC VIRAL HEPATITIS

Liver biopsy is the gold standard method for the grading and staging of chronic viral hepatitis, but optimal biopsy specimen size remains controversial. The aim of this study was to evaluate the quality of liver specimen (number of portal tracts) and to evaluate the impact of the number of portal tracts in the staging of chronic hepatitis. Material and Methods: 468 liver biopsies from consecutive patients with hepatitis C virus and hepatitis B virus infection from 2009 to 2010 were evaluated. Results: The length of fragment was less than 10 mm in 43 cases (9.3%), between 10 and 14 mm in 114 (24.3%), and ≥ 15 mm in 311 (64.4%); of these, in 39 (8.3%) cases were ≥ 20 mm. The mean representation of portal tracts was 17.6 ± 2.1 (5-40); in specimens ≥ 15 mm the mean portal tract was 13.5 ± 4.7 and in cases ≤ 15 mm was 11.4 ± 5.0 (p = 0.002). Cases with less than 11 portal tracts were associated with F3, and cases with 11 or more portal tracts with F2 (p = 0.001). Conclusion: this study demonstrated the good quality of liver biopsy and a relationship between the macroscopic size of the fragment and the number of portal tracts.

Labrea-like hepatitis in Vitoria, Espirito Santo state, Brazil: report of a case

A case of fulminat hepatitis with microvesicular steatosis resembling Labrea 's fever, diagnosed in Vitoria (ES) is reported. The 16 year old bcy presented with severe epistaxis, agitation, jaundice and hemorrhagic vomiting and died two days after admission to the emergency unit of the Vnivesity Hospital. The disease started five days before with fever, myalgias, dark urine and jaundice andprogressed withpsychic agitation, torpor and coma. The liver andspleen were notpalpable. HBsAg was negative in the serum. The autopsy showed acute hepatitis with tylic necrosis confluent in the midizonal and periportal areas with massive microvesicular steatosis in the remaining hepatocytes. Mononuclear cellspredominated in the exudate. The reticulum showed condensation in the necrotic areas without typical bands of collapse. The portal tracts were edematous with mononuclear infiltration and mild bile duct proliferation. Absence of cholestasis. Exceptfor the confluent midzonalandperiportal necrosis this case showed several clinical and morphological aspects of the Labreafever describedfrom the East Amazon, demonstrating that the anatomical picture of this disease probabty is not in related to afactor peculiar to the Amazon region.

A clinico-pathological study of 163 untreated cases of chronic hepatitis C

We performed a clinico-pathological study of 163 untreated cases of chronic hepatitis C. Eighty five percent of the patients were clinically asymptomatic and their physical examinations sbowed unremarkable or minimal changes at the time of the liver biopsy Liver function tests tended to present slight abnormalities, involving mild elevations of the activity of the aminotransferases and gamma-glutamil transferase levels. In spite of these mild abnormalities advanced chronic liver disease ivas histologically detected in eighty nine percent of the patients, mainly showing chronic active hepatitis. The most characteristic histological finding ivas an interlobular bile duct damage which correlated with the presence of tymphoid aggregates in the portal tracts and with the development of fibrosis.

Actinomycosis affecting the spinal cord: a case report

Actinomycosis is a rare, chronic, suppurative, granulomatous infection caused by a group of gram-positive anaerobic bacteria belonging to the natural flora of the oral cavity and gastrointestinal and urogenital tracts. It may involve several organs. This case study refers to pulmonary actinomycosis with chest wall involvement and cord compression in a 29-year-old male who presented with fever, cough, hemoptysis, neck pain, and paresis and plegia of the lower limbs of 5-month duration.

Temporal, spatial and spatiotemporal analysis of the occurrence of visceral leishmaniasis in humans in the City of Birigui, State of São Paulo, from 1999 to 2012

Introduction In 1999, Birigui and Araçatuba were the first municipalities in the State of São Paulo to present autochthonous cases of visceral leishmaniasis in humans (VLH). The aim of this study was to describe the temporal, spatial and spatiotemporal behaviors of VLH in Birigui. Methods Secondary data were obtained from the Notifiable Diseases Information System from 1999 to 2012. The incidence, mortality and case fatality rates by sex and age were calculated. The cases of VLH were geocoded and grouped according to census tracts. Local empirical Bayesian incidence rates were calculated. The existence of spatial and spatiotemporal clusters was investigated using SaTScan software. Results There were 156 confirmed cases of autochthonous VLH. The incidence rate was higher in the 0-4-year-old children, and the mortality and case fatality rates were higher in people aged 60 years and older. The peaks of incidence occurred in 2006 and 2011. The Bayesian rates identified the presence of VLH in all of the census tracts in the municipality; however, spatial and spatiotemporal clusters were found in the central area of the municipality. Conclusions Birigui, located in the Araçatuba region, has recently experienced increasing numbers of VLH cases; this increase is contrary to the behavior observed over the entire region, which has shown a decreasing trend in the number of VLH cases. The observations that the highest incidence is in children 0-4 years old and the highest mortality is in people 60 years and older are in agreement with the expected patterns of VLH.

Searching for therapeutic strategies in a mouse modelo of Machado-Joseph disease: targeting proteostases

Tese de Doutoramento em Ciências da Saúde

The role of the mammalian DNA end-processing enzyme polynucleotide kinase 3'-phosphatase in spinocerebellar ataxia Type 3 pathogenesis

DNA strand-breaks (SBs) with non-ligatable ends are generated by ionizing radiation, oxidative stress, various chemotherapeutic agents, and also as base excision repair (BER) intermediates. Several neurological diseases have already been identified as being due to a deficiency in DNA end-processing activities. Two common dirty ends, 3'-P and 5'-OH, are processed by mammalian polynucleotide kinase 3'-phosphatase (PNKP), a bifunctional enzyme with 3'-phosphatase and 5'-kinase activities. We have made the unexpected observation that PNKP stably associates with Ataxin-3 (ATXN3), a polyglutamine repeat-containing protein mutated in spinocerebellar ataxia type 3 (SCA3), also known as Machado-Joseph Disease (MJD). This disease is one of the most common dominantly inherited ataxias worldwide; the defect in SCA3 is due to CAG repeat expansion (from the normal 14-41 to 55-82 repeats) in the ATXN3 coding region. However, how the expanded form gains its toxic function is still not clearly understood. Here we report that purified wild-type (WT) ATXN3 stimulates, and by contrast the mutant form specifically inhibits, PNKP's 3' phosphatase activity in vitro. ATXN3-deficient cells also show decreased PNKP activity. Furthermore, transgenic mice conditionally expressing the pathological form of human ATXN3 also showed decreased 3'-phosphatase activity of PNKP, mostly in the deep cerebellar nuclei, one of the most affected regions in MJD patients' brain. Finally, long amplicon quantitative PCR analysis of human MJD patients' brain samples showed a significant accumulation of DNA strand breaks. Our results thus indicate that the accumulation of DNA strand breaks due to functional deficiency of PNKP is etiologically linked to the pathogenesis of SCA3/MJD.

First nesting record of the Bay-Capped Wren-Spinetail Spartonoica maluroides (Aves, Furnariidae) in Brazil, with nest and nestling descriptions and notes on breeding behavior

The only breeding record of Spartonoica maluroides (d'Orbigny & Lafresnaye, 1837) for Brazil is based on the observation of a fledgling in southern Rio Grande do Sul in January 1976. On 7 December 2005 we discovered a nest containing three nestlings at the southeastern end of Lagoa Pequena, municipality of Pelotas, Rio Grande do Sul. The nest was concealed at the base of a cavity in a Spartina densiflora (Poaceae) tussock located at the edge of a saltmarsh. The nest was built of fine pieces of dead Scirpus olneyi (Cyperaceae) and S. densiflora leaves firmly interlaced to the internal leaves of the tussock. Live leaves of S. densiflora lining the cavity comprised a substantial part of the nest's architecture, forming most of its upper lateral walls and roof. The lower section was more elaborate, resembling a deep cup and forming a distinct incubation chamber. Adults reached the nest's interior through an irregular apical opening amidst the leaves. The nest was 244 mm high and 140 mm wide. The incubation chamber had an external diameter of 138.5 mm, an internal diameter of 79.4 mm and was 86 mm deep. It was lined with fine leaves and white plant fibers. Nestlings were five to six days old. A total of 107 neossoptiles restricted to the capital, spinal and alar tracts were recorded in one nestling. The distribution of neossoptiles in the ocular region of S. maluroides forms a distinct pattern which can be typical of Furnariidae and related families. Two adults attended the nest, bringing small insects to the nestlings and removing fecal sacs. We recorded at least 74 visits to the nest during a ca. 6 h period during an afternoon. The average number of visits per hour was 12.8 ± 1.3. An adult bird spent on average 0.7 ± 0.56 minutes inside the nest attending nestlings. The nest remained unattended on average for 3.61 ± 3.13 minutes. The hour of the day had no influence on the amount of time spent by an adult in the nest or away from it. We returned to the area on 15 December 2005 and found the nest abandoned. Observations confirm that S. maluroides is a resident breeder in southern Brazil and that the saltmarshes of the Lagoa do Patos estuary are an important year-round habitat for the species. A nestling and the nest were collected to document the record.

Plastic ingestion by sea turtles in Paraíba State, Northeast Brazil

ABSTRACT Currently, plastics are recognized as a major pollutant of the marine environment, representing a serious threat to ocean wildlife. Here, we examined the occurrence and effects of plastic ingestion by sea turtles found stranded along the coast of Paraíba State, Brazil from August 2009 to July 2010. Ninety-eight digestive tracts were examined, with plastic found in 20 (20.4%). Sixty five percent (n = 13) of turtles with plastic in the digestive tract were green turtles (Chelonia mydas), 25% (n = 5) were hawksbills (Eretmochelys imbricata), and 10% (n = 2) were olive ridley (Lepidochelys olivacea). More plastic was found in the intestine (85%) than in other parts of the gastrointestinal tract. We observed complete blockage of the gastrointestinal tract due to the presence of plastic in 13 of the 20 turtles that had ingested plastic. No correlation was found between the curved carapace length (CCL) and the number or mass of the plastic ingested items. Significant differences were found between the intake of hard and soft plastic and the ingestion of white/transparent and colored plastic, with soft and white/transparent plastics being more commonly ingested. This study reveals the serious problem of plastic pollution to sea turtles at the area.

Patogenesis of pipe-stem fibrosis of the liver (experimental observation on murine Schistosomiasis)

Mice infected with 30 cercariae of Schistosoma mansoni developed portal and septal fibrosis due to the massive and concentrated deposition of eggs in the periportal areas which occurred following the 16th week after infection. The lesion resembled pipe-stem fibrosis seen in human hepatosplenic schistosomiasis in the following characters: portal fibrosis interconnecting portal spaces as well as portal spaces and central canals; portal inflammation; periovular granulomas; vascular obstruction and telangiectasia. The liver parenchyma maintained its normal architecture. Vascular injection techniques with Indian ink and vinylite revealed that the portal system developed numerous dilated collateral venules coming from the large and medium-sized portal branches, about 10 weeks after schistosome infection. The lodging of schistosome eggs into these collaterals resulted in granulomatous inflammation and fibrosis along all the portal tracts, thus forming the pipe-stem lesion. Although not readily demonstrable grossly, the pipe-stem fibrosis of murine schistosomiasis has many similarities with the human lesion and can be considered to have the same basic pathogenesis.

Overexpression of mutant ataxin-3 in mouse cerebellum induces ataxia and cerebellar neuropathology.

Machado-Joseph disease (MJD), also known as spinocerebellar ataxia type 3 (SCA3), is a fatal, dominant neurodegenerative disorder caused by the polyglutamine-expanded protein ataxin-3. Clinical manifestations include cerebellar ataxia and pyramidal signs culminating in severe neuronal degeneration. Currently, there is no therapy able to modify disease progression. In the present study, we aimed at investigating one of the most severely affected brain regions in the disorder-the cerebellum-and the behavioral defects associated with the neuropathology in this region. For this purpose, we injected lentiviral vectors encoding full-length human mutant ataxin-3 in the mouse cerebellum of 3-week-old C57/BL6 mice. We show that circumscribed expression of human mutant ataxin-3 in the cerebellum mediates within a short time frame-6 weeks, the development of a behavioral phenotype including reduced motor coordination, wide-based ataxic gait, and hyperactivity. Furthermore, the expression of mutant ataxin-3 resulted in the accumulation of intranuclear inclusions, neuropathological abnormalities, and neuronal death. These data show that lentiviral-based expression of mutant ataxin-3 in the mouse cerebellum induces localized neuropathology, which is sufficient to generate a behavioral ataxic phenotype. Moreover, this approach provides a physiologically relevant, cost-effective and time-effective animal model to gain further insights into the pathogenesis of MJD and for the evaluation of experimental therapeutics of MJD.

The spatial inequality of wage income in Barcelona Metropolitan Area

ABSTRACT The measure and estimation of income levels in Barcelona Metropolitan Area (BMA) goes back a long way. Using different approaches and focusing on different municipalities, there is a lot of work in the field. The majority of the literature has focused on the estimation of income levels using variables related to consumption. The empirical evidence on wage differentials has shown an important growth during 80’s and 90’s especially in United Kingdom and USA. Less is known on spatial distribution of inequality. This paper presents a new data set for analyzing spatial distribution of wage income. This data is obtained by matching Wage Structure Survey (WSS) with data from Census disaggregated by census tracts. In this way we have a unique data set with wage incomes for every census track for 36 municipalities belonging to BMA. We develop a descriptive analysis of spatial distribution, testing for spatial autocorrelation and use the family of Generalised Entropy Indices to measure inequality. Properties of the index allow us to decompose inequality into inter and intra-municipality measures. Since we have two cross-sectional data for WSS (1995-2002) we can also analyze the evolution of the inequality in this period of economic growth. Key words: spatial distribution of wages, spatial autocorrelation, inequality indices.