Is point of care testing for anaemia (HB) and microalbumin feasible in people with type 2 diabetes attending diabetes outpatient clinics?

Introduction: Diabetes is the major cause of chronic kidney disease (CKD) in Australia. Anaemia of CKD occurs earlier than in non-diabetics and is often insidious and undetected.

Aim: A large, prospective, single-centre study was undertaken to determine the feasibility of point of care testing (POCT) haemoglobin (Hb) and microalbumin in people with type 2 diabetes (T2DM) attending routine outpatient clinic appointments (OPC).

Method: Clinic nurses measured Hb and microalbumin using the HemoCue Haemoglobin Capillary Analyser and the HemoCue Urine Albumin Analyser (Medipac Scientific), respectively when they tested blood glucose, weight and blood pressure. The nurses were trained to use the analysers before the study commenced. Standard demographic data, duration of diabetes, treatment mode, and presence of complications, comorbidities, and HbA1c were ascertained from patients’ medical records.

Results: Five hundred and fifty-four (80%) patients were screened. The nurses were able to perform the tests competently but testing, especially microalbumin, was time-consuming. Patients’ mean age was 62 years (11 SD): 230 females, mean blood glucose (BG) 10 (3.9 SD) mmol/L, mean haemoglobin 127.2 (16.3 SD) g/L; mean microalbumin 47.8 (58.7 SD) mg/L: 324 were males, mean BG 10.2 (3.9 SD) mmol/L, mean Hb 138.6 (18.8 SD) gm/L, and mean microalbumin 67.9 (73.9 SD) mg/L. 27% of males and 22% of females were anaemic. Of those with anaemia, 27% of females and 29% of males had microalbuminuria.

Conclusions: POCT is feasible in routine outpatient clinics but is time-consuming. One in four T2DM attending OPC were anaemic. POCT Hb testing in OPC is feasible and could identify T2DM who need full haematological assessment.

Little evidence that condition, stress indicators, sex ratio, or homozygosity are related to landscape or habitat attributes in declining woodland birds

Habitat loss, fragmentation and degradation are drivers of major declines in biodiversity and species extinctions. The actual causes of species population declines following habitat change are more difficult to discern and there is typically high covariation among the measures used to infer the causes of decline. The causes of decline may act directly on individual fitness and survival, or through disruption of population processes. We examined the relationships among configuration, extent and status of native vegetation and three commonly used indicators of individual body condition and chronic stress (haemoglobin level, haematocrit, residual body mass condition index) in 13 species of woodland-dependent birds in south-eastern Australia. We also examined two measures of changes to population processes (sex ratio and individual homozygosity) in ten species and alleic richness in five species. We found little support for relationships between site or landscape characteristics and individual or population response variables, notwithstanding that our simulations showed we had sufficient power to detect relatively small effects. We discuss possible causes of the absence of detectable habitat effects in this system and the implications for the usefulness of individual body condition and easily measured haematological indices as indicators of the response of avian populations to habitat change. © 2012 The Authors.

Helicobacter pylori-associated hypochlorhydria in children, and development of iron deficiency

Aims Acute Helicobacter pylori infection is associated with transient hypochlorhydria. In H pylori-associated atrophy, hypochlorhydria has a role in iron deficiency (ID) through changes in the physiology of iron-complex absorption. The aims were to evaluate the association between H pylori-associated hypochlorhydria and ID in children. Methods Symptomatic children (n=123) were prospectively enrolled. Blood, gastric juice and gastric biopsies were taken, respectively, for haematological analyses, pH assessment and H pylori determination, and duodenal biopsies for exclusion of coeliac disease. Stool samples were collected for parasitology/microbiology. Thirteen children were excluded following parasitology and duodenal histopathology, and five due to impaired blood analysis. Results Ten children were hypochlorhydric (pH>4) and 33 were H pylori positive. In H pylori-positive children with pH>4 (n=6) serum iron and transferrin saturation levels % were significantly lower (p<0.01) than H pylori-positive children with pH≤4. No differences in ferritin, or total iron binding capacity, were observed. In H pylori-negative children with pH>4, iron and transferrin saturation were not significantly different from children with pH≤4. Conclusions Low serum iron and transferrin in childhood H pylori infection is associated with hypochlorhydria. In uninfected children, hypochlorhydria was not associated with altered serum iron parameters, indicating a combination of H pylori infection and/or inflammation, and hypochlorhydria has a role in the aetiology of ID. Although H pylori-associated hypochlorhydria is transient during acute gastritis, this alters iron homeostasis with clinical impact in developing countries with a high H pylori prevalence.

ETV6 (TEL1) in blood cell development and malignancy

 The ETV6 (TEL1) gene encodes a member of the ETS family of transcriptional regulators. It represents one of the most frequently disrupted genes in haematological malignancies, with over 50 different translocations described. Moreover, deletion, silencing or truncating mutations have also been reported, suggesting a potential tumor suppressor function. Recent studies have shown that ETV6 plays a broad and complex role in early hematopoiesis, impacting on the development of multiple lineages, providing new insights into how its perturbation contributes to disease.

Memory improvements in elderly women following 16 weeks treatment with a combined multivitamin, mineral and herbal supplement: a randomized controlled trial.

RATIONALE: There is potential for multivitamin supplementation to improve cognition in the elderly. This randomized, double-blind, placebo-controlled trial was conducted to investigate the effects of 16 weeks multivitamin supplementation (Swisse Women's 50+ Ultivite ®) on cognition in elderly women. METHODS: Participants in this study were 56 community dwelling, elderly women, with subjective complaints of memory loss. Cognition was assessed using a computerized battery of memory and attention tasks designed to be sensitive to age-related declines to fluid intelligence, and a measure of verbal recall. Biochemical measures of selected nutrients, homocysteine, markers of inflammation, oxidative stress, and blood safety parameters were also collected. All cognitive and haematological parameters were assessed at baseline and 16 weeks post-treatment. RESULTS: The multivitamin improved speed of response on a measure of spatial working memory, however benefits to other cognitive processes were not observed. Multivitamin supplementation decreased levels of homocysteine and increased levels of vitamin B(6) and B(12), with a trend for vitamin E to increase. There were no hepatotoxic effects of the multivitamin formula indicating this supplement was safe for everyday usage in the elderly. CONCLUSION: Sixteen weeks ssupplementation with a combined multivitamin, mineral and herbal formula may benefit working memory in elderly women at risk of cognitive decline.

Invasive infections due to filamentous fungi other than Aspergillus: epidemiology and determinants of mortality

The epidemiology of invasive fungal disease (IFD) due to filamentous fungi other than Aspergillus may be changing. We analysed clinical, microbiological and outcome data in Australian patients to determine the predisposing factors and identify determinants of mortality. Proven and probable non-Aspergillus mould infections (defined according to modified European Organization for Research and Treatment of Cancer/Mycoses Study Group criteria) from 2004 to 2012 were evaluated in a multicentre study. Variables associated with infection and mortality were determined. Of 162 episodes of non-Aspergillus IFD, 145 (89.5%) were proven infections and 17 (10.5%) were probable infections. The pathogens included 29 fungal species/species complexes; mucormycetes (45.7%) and Scedosporium species (33.3%) were most common. The commonest comorbidities were haematological malignancies (HMs) (46.3%) diabetes mellitus (23.5%), and chronic pulmonary disease (16%); antecedent trauma was present in 21% of cases. Twenty-five (15.4%) patients had no immunocompromised status or comorbidity, and were more likely to have acquired infection following major trauma (p <0.01); 61 (37.7%) of cases affected patients without HMs or transplantation. Antifungal therapy was administered to 93.2% of patients (median 68 days, interquartile range 19-275), and adjunctive surgery was performed in 58.6%. The all-cause 90-day mortality was 44.4%; HMs and intensive-care admission were the strongest predictors of death (both p <0.001). Survival varied by fungal group, with the risk of death being significantly lower in patients with dematiaceous mould infections than in patients with other non-Aspergillus mould infections. Non-Aspergillus IFD affected diverse patient groups, including non-immunocompromised hosts and those outside traditional risk groups; therefore, definitions of IFD in these patients are required. Given the high mortality, increased recognition of infections and accurate identification of the causative agent are required.

Incidência de infecção por Legionella pneumophila em pacientes que internaram no HCPA com pneumonia adquirida na comunidade

Introdução: O diagnóstico microbiológico da infecção por Legionella é complexo, pois a bactéria não é visualizada à coloração de Gram no escarro, e sua cultura não é realizada na maioria dos laboratórios clínicos. A imunofluorescência direta nas secreções respiratórias tem baixa sensibilidade, em torno de 40% e a técnica da “PCR” não é ainda recomendada para o diagnóstico clínico (CDC, 1997). A detecção de anticorpos no soro é a técnica mais utilizada, e o critério definitivo é a soroconversão para no mínimo 1:128, cuja sensibilidade é de 70 a 80% (Edelstein, 1993). Como critérios diagnósticos de possível pneumonia por Legionella, eram utilizados: título único de anticorpos a L pneumophila positivo na diluição 1:256, em paciente com quadro clínico compatível (CDC, 1990) e o achado de antígeno a Legionella na urina (WHO, 1990). Nos últimos anos, porém, com o uso crescente do teste de antigenúria, foram detectados casos de pneumonia por Legionella, que não eram diagnosticados por cultura ou sorologia, tornando-o método diagnóstico de certeza para o diagnóstico de pneumonia por Legionella (CDC, 1997). Por sua fácil execução, resultado imediato, e alta sensibilidade - de 86% a 98% (Kashuba & Ballow, 1986; Harrison & Doshi, 2001), tem sido recomendado para o diagnóstico das PAC que necessitam internação hospitalar (Mulazimoglu & Yu, 2001; Gupta et al., 2001; Marrie, 2001), especialmente em UTI (ATS, 2001). Vários estudos documentaram baixo valor preditivo positivo do título único positivo de 1:256, tornando-o sem valor para o diagnóstico da pneumonia por Legionella, exceto, talvez, em surtos (Plouffe et al., 1995). Outros detectaram alta prevalência de anticorpos positivos na diluição 1:256 na população, em pessoas normais (Wilkinson et al., 1983; Nichol et al., 1991). A partir de 1996, o CDC de Atlanta recomendou que não seja mais utilizado o critério de caso provável de infecção por Legionella pneumophila por título único de fase convalescente ≥1:256, por falta de especificidade(CDC, 1997). A pneumonia por Legionella é raramente diagnosticada, e sua incidência é subestimada. Em estudos de PAC, a incidência da pneumonia por Legionella nos EUA, Europa, Israel e Austrália, foi estimada entre 1% a 16% (Muder & Yu, 2000). Nos EUA, foi estimado que cerca de 8 000 a 23 000 casos de PAC por Legionella ocorrem anualmente, em pacientes que requerem hospitalização (Marston et al., 1994 e 1977). No Brasil, a incidência de PAC causadas por Legionella em pacientes hospitalizados é tema de investigação pertinente, ainda não relatado na literatura. Objetivo: detectar a incidência de pneumonias causadas por Legionella pneumophila sorogrupos 1 a 6, em pacientes que internaram no Hospital de Clínicas de Porto Alegre por PAC, por um ano. Material e Métodos: o delineamento escolhido foi um estudo de coorte (de incidência), constituída por casos consecutivos de pneumonia adquirida na comunidade que internaram no HCPA de 19 de julho de 2000 a 18 de julho de 2001. Para a identificação dos casos, foram examinados diariamente o registro computadorizado das internações hospitalares, exceto as internações da pediatria e da obstetrícia, sendo selecionados todos os pacientes internados com o diagnóstico de pneumonia e de insuficiência respiratória aguda. Foram excluídos aqueles com menos de 18 anos ou mais de 80 anos; os procedentes de instituições, HIV-positivos, gestantes, pacientes restritos ao leito; e portadores de doença estrutural pulmonar ou traqueostomias. Foram excluídos os pacientes que tivessem tido alta hospitalar nos últimos 15 dias, e aqueles já incluídos no decorrer do estudo. Os pacientes selecionados foram examinados por um pesquisador, e incluídos para estudo se apresentassem infiltrado ao RX de tórax compatível com pneumonia, associado a pelo menos um dos sintomas respiratórios maiores (temperatura axilar > 37,8ºC, tosse ou escarro; ou dois sintomas menores (pleurisia, dispnéia, alteração do estado mental, sinais de consolidação à ausculta pulmonar, mais de 12 000 leucócitos/mm3). O estudo foi previamente aprovado pela Comissão de Ética em Pesquisa do HCPA. Os pacientes eram entrevistados por um pesquisador, dando seu consentimento por escrito, e então seus dados clínicos e laboratoriais eram registrados em protocolo individual. Não houve interferência do pesquisador, durante a internação, exceto pela coleta de urina e de sangue para exame laboratoriais específicos da pesquisa. Os pacientes eram agendados, no ambulatório de pesquisa, num prazo de 4 a 12 semanas após sua inclusão no estudo, quando realizavam nova coleta de sangue, RX de tórax de controle, e outros exames que se fizessem necessários para esclarecimento diagnóstico.Todos os pacientes foram acompanhados por 1 ano, após sua inclusão no estudo.Foram utilizadas a técnica de imunofluorescência indireta para detecção de anticorpos das classes IgG, IgM e IgA a Legionella pneumophila sorogrupos 1 a 6 no soro, em duas amostras, colhidas, respectivamente, na 1ª semana de internação e depois de 4 a 12 semanas; e a técnica imunológica por teste ELISA para a detecção do antígeno de Legionella pneumophila sorogrupo 1 na urina, colhida na primeira semana de internação. As urinas eram armazenadas, imediatamente após sua coleta, em freezer a –70ºC, e depois descongeladas e processadas em grupos de cerca de 20 amostras. A imunofluorescência foi feita no laboratório de doenças Infecciosas da Universidade de Louisville (KY, EUA), em amostras de soro da fase aguda e convalescente, a partir da diluição 1:8; e a detecção do antígeno de Legionella pneumophila sorogrupo 1, nas amostras de urina, foi realizada no laboratório de pesquisa do HCPA, pelos investigadores, utilizando um kit comercial de teste ELISA fabricado por Binax (Binax Legionella Urinary Enzyme Assay, Raritan, EUA). As urinas positivas eram recongeladas novamente, para serem enviadas para confirmação no mesmo laboratório americano, ao fim do estudo. Foram adotados como critérios definitivos de infecção por Legionella pneumophila sorogrupos 1 a 6, a soroconversão (elevação de 4 vezes no título de anticorpos séricos entre o soro da fase aguda e da fase convalescente para no mínimo 1:128); ou o achado de antígeno de L pneumophila sorogrupo 1 na urina não concentrada, numa razão superior a 3, conforme instruções do fabricante e da literatura.Os pacientes foram classificados, de acordo com suas características clínicas, em 1º) portadores de doenças crônicas (doenças pulmonares, cardíacas, diabete mellitus, hepatopatias e insuficiência renal); 2º) portadores de doenças subjacentes com imunossupressão; 3º) pacientes hígidos ou com outras doenças que não determinassem insuficiência orgânica. Imunossupressão foi definida como esplenectomia, ser portador de neoplasia hematológica, portador de doença auto-imune, ou de transplante; ou uso de medicação imunossupressora nas 4 semanas anteriores ao diagnóstico (Yu et al., 2002b); ou uso de prednisolona 10 mg/dia ou equivalente nos últimos 3 meses (Lim et al., 2001). As características clínicas e laboratoriais dos pacientes que evoluíram ao óbito por pneumonia foram comparados àquelas dos pacientes que obtiveram cura. Para a análise das variáveis categóricas, utilizou-se o teste qui-quadrado de Pearson ou teste exato de Fisher. Para as variáveis numéricas contínuas, utilizou-se o teste “t“ de Student. Um valor de p< 0,05 foi considerado como resultado estatisticamente significativo (programas SPSS, versão 10). Foi calculada a freqüência de mortes por pneumonia na população estudada, adotando-se a alta hospitalar como critério de cura. Foi calculada a incidência cumulativa para pneumonia por Legionella pneumophila sorogrupos 1 a 6, em um hospital geral, no período de 1 ano. Resultados: durante um ano de estudo foram examinados 645 registros de internação, nos quais constavam, como motivo de baixa hospitalar, o diagnóstico de pneumonia ou de insuficiência respiratória aguda; a maioria desses diagnósticos iniciais não foram confirmados. Desses 645 pacientes, foram incluídos no estudo 82 pacientes, nos quais os critérios clínicos ou radiológicos de pneumonia foram confirmados pelos pesquisadores. Durante o acompanhamento desses pacientes, porém, foram excluídos 23 pacientes por apresentarem outras patologias que mimetizavam pneumonia: DPOC agudizado (5), insuficiência cardíaca (3), tuberculose pulmonar (2), colagenose (1), fibrose pulmonar idiopática (1), edema pulmonar em paciente com cirrose (1), somente infecçâo respiratória em paciente com sequelas pulmonares (4); ou por apresentarem critérios de exclusão: bronquiectasias (4), HIV positivo (1), pneumatocele prévia (1). Ao final, foram estudados 59 pacientes com pneumonia adquirida na comunidade, sendo 20 do sexo feminino e 39 do sexo masculino, com idade entre 24 e 80 anos (média de 57,6 anos e desvio padrão de ±10,6). Tivemos 36 pacientes com doenças subjacentes classificadas como “doenças crônicas”, dos quais 18 pacientes apresentavam mais de uma co-morbidade, por ordem de prevalência: doenças pulmonares, cardíacas, diabete mellitus, hepatopatias e insuficiência renal; neoplasias ocorreram em 9 pacientes, sendo sólidas em 7 pacientes e hematológicas em 2. Dos 59 pacientes, 61% eram tabagistas e 16,9%, alcoolistas. Do total, 10 pacientes apresentavam imunossupressão. Dos demais 13 pacientes, somente um era previamente hígido, enquanto os outros apresentavam tabagismo, sinusite, anemia, HAS, gota, ou arterite de Takayasu. A apresentação radiológica inicial foi broncopneumonia em 59,3% dos casos; pneumonia alveolar ocorreu em 23,7% dos casos, enquanto ambos padrões ocorreram em 15,2% dos pacientes. Pneumonia intersticial ocorreu em somente um caso, enquanto broncopneumonia obstrutiva ocorreu em 5 pacientes (8,5%). Derrame pleural ocorreu em 22% dos casos, e em 21 pacientes (35%) houve comprometimento de mais de um lobo ao RX de tórax. Foram usados beta-lactâmicos para o tratamento da maioria dos pacientes (72,9%9). A segunda classe de antibióticos mais usados foi a das fluoroquinolonas respiratórias, que foram receitadas para 23 pacientes (39,0%), e em 3º lugar, os macrolídeos, usados por 11 pacientes (18,6%). Apenas 16 pacientes não usaram beta-lactâmicos, em sua maioria recebendo quinolonas ou macrolídeos. Dos 43 pacientes que usaram beta-lactâmicos, 25 não usaram nem macrolídeos, nem quinolonas. Em 13 pacientes as fluoroquinolonas respiratórias foram as únicas drogas usadas para o tratamento da pneumonia. Do total, 8 pacientes foram a óbito por pneumonia; em outros 3 pacientes, o óbito foi atribuído a neoplasia em estágio avançado. Dos 48 pacientes que obtiveram cura, 33 (68,7%) estavam vivos após 12 meses. Os resultados da comparação realizada evidenciaram tendência a maior mortalidade no sexo masculino e em pacientes com imunossupressão, porém essa associação não alcançou significância estatística. Os pacientes que usaram somente beta-lactâmicos não apresentaram maior mortalidade do que os pacientes que usaram beta-lactâmicos associados a outras classes de antibióticos ou somente outras classes de antibióticos. Examinando-se os pacientes que utiizaram macrolídeos ou quinolonas em seu regime de tratamento, isoladamente ou combinados a outros antibióticos, observou-se que também não houve diferença dos outros pacientes, quanto à mortalidade. Os pacientes com padrão radiológico de pneumonia alveolar tiveram maior mortalidade, e essa diferença apresentou uma significância limítrofe (p= 0,05). Nossa mortalidade (11,9%) foi similar à de Fang et al. (1990), em estudo clássico de 1991 (13,7%); foi também similar à média de mortalidade das PAC internadas não em UTI (12%), relatada pela ATS, no seu último consenso para o tratamento empírico das PAC (ATS, 2001). Foram detectados 3 pacientes com pneumonia por Legionella pneumophila sorogrupo 1 na população estudada: 2 foram diagnosticados por soroconversão e por antigenúria positiva, e o 3º foi diagnosticado somente pelo critério de antigenúria positiva, tendo sorologia negativa, como alguns autores (McWhinney et al., 2000). Dois pacientes com PAC por Legionella não responderam ao tratamento inicial com beta-lactâmicos, obtendo cura com levofloxacina; o 3º paciente foi tratado somente com betalactâmicos, obtendo cura. Conclusões: A incidência anual de PAC por Legionella pneumophila sorogrupos 1 a 6, no HCPA, foi de 5,1%, que representa a incidência anual de PAC por Legionella pneumophila sorogrupos 1 a 6 em um hospital geral universitário. Comentários e Perspectivas: Há necessidade de se empregar métodos diagnósticos específicos para o diagnóstico das pneumonias por Legionella em nosso meio, como a cultura, a sorologia com detecção de todas as classes de anticorpos, e a detecção do antígeno urinário, pois somente com o uso simultâneo de técnicas complementares pode-se detectar a incidência real de pneumonias causadas tanto por Legionella pneumophila, como por outras espécies. A detecção do antígeno de Legionella na urina é o teste diagnóstico de maior rendimento, sendo recomendado seu uso em todas as PAC que necessitarem internação hospitalar (Mulazimoglu & Yu, 2001; Gupta et al., 2001); em todos os pacientes com PAC que apresentarem fatores de risco potenciais para legionelose (Marrie, 2001); e para o diagnóstico etiológico das pneumonias graves (ATS, 2001). Seu uso é indicado, com unanimidade na literatura, para a pesquisa de legionelose nosocomial e de surtos de legionelose na comunidade.

Estudo sobre a participação de rodedores na cadeia de transmissão de Leishmania infantum (Protozoa: Trypanosomatidae) no Rio Grande do Norte

American visceral leishmaniasis is a zoonosis caused by Leishmania infantum and transmitted by the bite of the sand flies Lutzomia longipalpis.The main domestic reservoir is the dog, while foxes and opposums are the known wild reservoirs. However, identification of natural infections with L. infantum in rodents appears for need of investigating the participation of these rodents how source of infection of the parasite. In the present work the Leishmania infantum infection was investigated in rodents captured in Rio Grande do Norte, aiming at to offer subsidies to the understanding of the epidemic chains of LVA in the State. Thirteen Galea spixii were distributed in four groups, being G1 the group control with four animals and the others, G2, G3 and G4, with three animals each. Those animals were intraperitoneally inoculated with 107 promastigotas of L. infantum and accompanied for, respectively, 30, 90 and 180 days. Weekly the animals were monitored as for the corporal weight and rectal temperature. At the end of each stipulated period the animals were killed. Blood were used for determination of the parameters biochemical and haematological, PCR, ELISA, microscopic examination and cultivation in NNN medium. Liver, spleen and lymph node were used in Giemsa-stained impression and cultivation in NNN medium. Liver and spleen fragments were still used in PCR and histopathological, respectively. At the same time 79 rodents of the species Rattus rattus, Bolomys lasiurus, Oligoryzomys nigripis, Oryzomys subflavus and Trichomys apereoides were captured in the Municipal districts of Brejinho, Campo Grande, Coronel Ezequiel, Passa e Fica and Vázea for identification of natural infection with L. infantum. Evidence of infection was checked by direct examination of Giemsa-stained impression of liver, spleen and blood and culture of these tissues in NNN medium. Antibodies were researched by ELISA. They were not found differences among the weigh corporal final, rectal temperature and biochemical and haematological parameters of the Galea spixii controls and infected. The rectal temperature of the animals varied from 36OC to 40OC. For the first time values of the haematocrit (33,6% to 42,8%), hemoglobin (10,2 to 14,5g/dl), erythrocyts number (4,67x106 to 6,90x106/mm3), total leukocytes (0,9x103 to 9,2x103/mm3), platelets (49x103 to 509x103/mm3) total proteins (1,56 to 6,06 g/dl), albumin (1,34 to 3,05 g/dl) and globulins (0,20 to 3,01 g/dl) of the Galea spixii were determined. The lymphocytes were the most abundant leucocytes. Infection for L. infantum was diagnosed in two animals euthanasied 180 days after the infection. In one of the animals was also identified antibodies anti-Leishmania. The parasite was not found in none of the five other species of rodents captured. Galea spixii are resistant to the infection for L. infantum and they are not good models for the study for visceral leishmaniose, although they can act as infection sources. More studies are necessary to determine the paper of the rodents in the epidemic chain of transmission of the visceral leishmaniose in the State of Rio Grande do Norte

Influência de diferentes níveis de vitamina C na ração sobre parâmetros hematológicos de alevinos de Piaractus mesopotamicus Holmberg (Osteichthyes, Characidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo de polimorfismos em genes relacionados ao metabolismo do ácido fólico e sua associação com o desenvolvimento de fendas orais não-sindrômicas

The congenital facial clefts are characterized by incomplete formation of the structures that separate the oral and nasal cavity. It is known that several environmental and genetic factors are involved in its development, among these, polymorphisms associated with folic acid metabolism have been investigated. In this sense, the objective was to observe the frequency of polymorphisms C677T and A1298C methylenetetrahydrofolate reductase gene (MTHFR), methionine synthase A2756G of (MTR), A66G of methionine synthase reductase (MTRR) A80G and the reduced folate carrier (RFC1) in patients with non-syndromic oral clefts, trying to match them with their development. Methods: We studied 140 patients with non-syndromic oral clefts and their mothers and 175 control subjects with their mothers, who underwent a questionnaire to obtain family information. Were collecting blood for DNA extraction from patients and their mothers to identify the genotypes of both by PCRRFLP, in addition to carrying out the determination of glucose, AST, ALT and serum creatinine, folic acid and vitamin B12 Serum and plasma homocysteine, and the hemogram. Results: Most patients have cleft lip and palate (55.8%), followed by isolated cleft palate (24.2%) and cleft lip (20%). Regarding gender, 62% of patients were male and 48% female and, after subdivision of the type of screwdriver according to sex was found a prevalence of males in the cracks of the type lip and palate (69 %) and lip (69.2%) and in the case of cleft palate was a female predominance (59%). The average concentration of serum folate in the group of mothers of cleft patients was significantly lower (13.8 ± 2.4 ng / mL) compared with the group of mothers of control subjects (18.8 ± 3.4 ng / mL) This was also observed for the group of cleft children as compared to controls, the dosage of folic acid had a significant difference with values of 15.6 ± 0.6 (ng / mL) and 17.9 ± 0.6 (ng / mL), respectively. For the biochemical measurements of glucose, AST, ALT and creatinine were not statistically different, nor was observed for haematological parameters performed. In assessing the frequency of polymorphisms C677T and A1298C MTHFR, A2756G MTR, MTRR A66G and A80G of the RFC1 there was no statistically significant difference in genotype distribution between cases and controls both for mothers and in the cleft. Conclusion: Although not observed association of polymorphisms with the development of cracks, the decrease in serum folate in the group of cleft patients and their mothers may reflect a disturbance in the metabolism of this metabolite, necessitating further studies such as studies methylation and expression to further elucidate the involvement of folate in the development of oral clefts

Haematologic and immunologic parameters of bullfrogs, Lithobates catesbeianus, fed probiotics

The effects of two probiotics (P(1) - Lactobacillus acidophilus, Bifidobacterium bifidum and Enterococcus faecium and P(2) - Bacillus subtilis) supplemented to commercial feed (40% crude protein) on the haematological and immunological parameters of the bullfrog Lithobates catesbeianus were studied. Two doses of each probiotic (5 and 10 g kg-1 of food) were added to the diets and fed to frogs, totalling five treatments over 112 days. Haematological analyses consisted of total and differential leucocyte counts, erythrocyte and thrombocyte counts, haematocrit, haemoglobin levels and RBC indices (mean corpuscular volume, mean corpuscular haemoglobin - and mean corpuscular haemoglobin concentration) and the immunological parameters included phagocytic capacity and phagocytic index of peritoneal phagocytes. The results showed that the probiotics did not significantly influence any of the haematological parameters measured. However, immunological assays showed that the probiotics had an immunostimulating effect. The greatest effects were seen with probiotic P(1) fed at a dose of 10 g kg-1 of diet and probiotic P(2) fed at 5 g kg-1 of diet.

Granulocytic sarcoma of the small intestine with CBF beta/MYH11 fusion gene: report of an aleukaemic case and review of the literature

Granulocytic sarcomas (GS) are rare extramedullary tumours composed of immature myeloid cells. Inversion of chromosome 16 [inv(16)] is a cytogenetic marker for M4Eo subtype of acute myeloid leukaemia (AML). The possibility of an association between the development of granulocytic sarcoma of the small intestine (GSSI) and the M4Eo subtype of AML was suggested in nine previous case reports.Here we report an aleukaemic case of GSSI with inv(16) and its molecular equivalent, the CBFbeta/MYH11 fusion gene, detected by reverse transcriptase-polymerase chain reaction (RT-PCR), that after treatment with conventional AML chemotherapy followed by autologous bone marrow transplantation, achieved complete haematological and molecular remission on bone marrow examination. After chemotherapy, a thickened ileum wall positive for CBFbeta/MYH11 on tumour mass samples was still observed on computed tomography (CT) studies, raising the question of residual GS representing a reservoir of malignant cells. This case demonstrates the critical need of multidisciplinary diagnosis and follow-up of this entity combining immunopathologic, cytogenetic and molecular studies, reinforcing the potentiality of risk-adapted therapy strategies, as it is increasingly claimed for patients with overt AML. (C) 2003 Elsevier Ltd. All rights reserved.

Clinical, histological and immunophenotypic findings in a mare with a mammary lymphoma associated with anaemia and pruritus

An 18-year-old Quarter Horse mare presented with an 18 week history of dermatological lesions characterised by recurrent urticaria and recent signs of enlarged mammary glands, pyrexia, alopecia and intense pruritus. Haematological and serum biochemical analyses revealed anaemia, leucocytosis, thrombocytopenia and hyperfibrinogenaemia. Mammary discharge was evaluated and revealed neutrophilia. Cytological examinations of fine needle aspirates from the parotid lymph nodes were normal. Common causes of anaemia were ruled out by specific tests. The initial diagnosis was mastitis and 2 weeks later the animal returned to the hospital with worsening of clinical signs, intermittent fever, intense pruritus and generalised alopecia. Blood samples were collected for haematological and serum biochemical analyses, and the mammary gland and parotid lymph nodes were biopsied. Rectal palpation revealed a large irregular mass near the left kidney with adhesions of surrounding structures. The animal died before the laboratory results were ready. This report details the clinical, histological and immunophenotypic findings of a case of large lymphoma in a mare.

Field study on nematode resistance in Nelore-breed cattle

The present study evaluated Nelore cattle with different degrees of resistance to natural infections by gastrointestinal nematodes. One hundred weaned male cattle, 11-12 months of age, were kept on the same pasture and evaluated from October 2003 to February 2004. Faecal and blood samples were collected for parasitological, haematological and immunological tests. In February 2004, the 10 most resistant and the 10 most susceptible animals were selected based on individual means of nematode faecal egg counts (FEC). Such animals were slaughtered for worm burden determination and nematode species identification. The repeatability estimates for FEC (+/- S.D.), log-transformed FEC and packed-cell volume (PCV) in all animals were 0.3 (+/- 0.05), 0.26 (+/- 0.04) and 0.42 (+/- 0.05), respectively. The resistant group showed lower FEC and worm burdens than the susceptible group (P < 0.05). There were no significant differences between groups regarding mean body weight, weight gain, PCV and total serum protein values (P > 0.05). The resistant group showed higher total serum IgE levels (P < 0.05) and higher mean eosinophil blood counts. However, the latter was statistically significant only 42 days after the beginning of the study. Nematodes Cooperia punctata and Haemonchus placei were predominant and the correlation between Cooperia and Haemonchus burdens was 0.64 (P < 0.05), which indicated that animals presenting increased numbers of one of those genera probably had increased numbers of the other. The current study provides further evidence of IgE active role in nematode immunity and suggests that total serum IgE level might serve as an additional marker to select Nelore cattle that are responsive to H. placei and C. punctata infections. (C) 2007 Elsevier B.V. All rights reserved.

Effects of inhibition gastric acid secretion on arterial acid-base status during digestion in the toad Bufo marinus

Digestion affects acid-base status, because the net transfer of HCl from the blood to the stomach lumen leads to an increase in HCO3- levels in both extra- and intracellular compartments. The increase in plasma [HCO3-], the alkaline tide, is particularly pronounced in amphibians and reptiles, but is not associated with an increased arterial pH, because of a concomitant rise in arterial Pco(2) caused by a relative hypoventilation. In this study, we investigate whether the postprandial increase in Paco(2) of the toad Bufo marinus represents a compensatory response to the increased plasma [HCO3-] or a state-dependent change in the control of pulmonary ventilation. To this end, we successfully prevented the alkaline tide, by inhibiting gastric acid secretion with omeprazole, and compared the response to that of untreated toads determined in our laboratory during the same period. In addition, we used vascular infusions of bicarbonate to mimic the alkaline tide in fasting animals. Omeprazole did not affect blood gases, acid-base and haematological parameters in fasting toads, but abolished the postprandial increase in plasma [HCO3-] and the rise in arterial Pco(2) that normally peaks 48 h into the digestive period. Vascular infusion of HCO3-, that mimicked the postprandial rise in plasma [HCO3-], led to a progressive respiratory compensation of arterial pH through increased arterial Pco(2) Thus, irrespective of whether the metabolic alkalosis is caused by gastric acid secretion in response to a meal or experimental infusion of bicarbonate, arterial pH is being maintained by an increased arterial Pco(2). It seems, therefore, that the elevated Pco(2), occuring during the postprandial period, constitutes of a regulated response to maintain pH rather than a state-dependent change in ventilatory control. (C) 2003 Elsevier B.V. All rights reserved.