A nested-PCR with an Internal Amplification Control for the detection and differentiation of Bartonella henselae and B. clarridgeiae: An examination of cats in Trinidad

BACKGROUND: Bartonella species are bacterial blood parasites of animals capable of causing disease in both animals and man. Cat-Scratch Disease (CSD) in humans is caused mainly by Bartonella henselae and is acquired from the cat, which serves as a reservoir for the bacteria. A second species, B. clarridgeiae is also implicated in the disease. Diagnosis of Bartonellosis by culture requires a week or more of incubation on enriched media containing blood, and recovery is often complicated by faster growing contaminating bacteria and fungi. PCR has been explored as an alternative to culture for both the detection and species identification of Bartonella, however sensitivity problems have been reported and false negative reactions due to blood inhibitors have not generally been addressed in test design. METHODS: A novel, nested-PCR was designed for the detection of Bartonella henselae and B. clarridgeiae based on the strategy of targeting species-specific size differences in the 16S-23S rDNA intergenic regions. An Internal Amplification Control was used for detecting PCR inhibition. The nested-PCR was utilized in a study on 103 blood samples from pet and stray cats in Trinidad. RESULTS: None of the samples were positive by primary PCR, but the Nested-PCR detected Bartonella in 32/103 (31%) cats where 16 were infected with only B. henselae, 13 with only B. clarridgeiae and 3 with both species. Of 22 stray cats housed at an animal shelter, 13 (59%) were positive for either or both species, supporting the reported increased incidence of Bartonella among feral cats. CONCLUSION: The usefulness of a single PCR for the detection of Bartonella henselae and B. clarridgeiae in the blood of cats is questionable. A nested-PCR offers increased sensitivity over a primary PCR and should be evaluated with currently used methods for the routine detection and speciation of Bartonella henselae and B. clarridgeiae. In Trinidad, B. henselae and B. clarridgeiae are the predominant species in cats and infection appears highest with stray cats, however B. clarridgeiae may be present at levels similar to that of B. henselae in the pet population

Efficacy of prophylaxis and genotype-phenotype correlation in patients with severe Factor X deficiency in Iran

We aimed to evaluate the effect of regular prophylaxis with a Factor X (FX) concentrate for patients with severe FXD in Iran and to assess the correlation of the genotype and phenotype in these patients. Ten patients with severe FXD (FX activity <1%) were enrolled and characterized during 2010-2011. Prophylaxis with 20\u2003IU FX P Behring per kg body weight was administered once a week. FX levels, were monitored at baseline, 15 and 30\u2003min, 1, 3, 6, 12, 24, 48, 72 and 96\u2003h after starting prophylaxis. All patients were followed for 1\u2003year. The mean age of the patients was 15\u2003\ub1\u20037.8\u2003years (age range of: 6-27\u2003years). One patient had anaphylactic reaction after the first infusion, and the treatment was stopped. During one-year follow-up after starting prophylaxis, no bleeding symptoms occurred in any patient who tolerated and remained on the prophylaxis programme and all of them had a FX level of 1% or above. The maximum level of FX activity has been observed at 15\u2003min after starting prophylaxis. A level of 1.5-3.5% was detected after 96\u2003h. Homozygous mutations p.Arg40Thr (Arg-1Thr), p.Gly51Arg and p.Glu69Lys were detected in patients with intracranial haemorrhage. In our patients, significant decrease in symptoms without any complication after administration of FX, was demonstrated in all except one patient who had an anaphylactic reaction. It seems that the dose of 20\u2003IU\u2003kg(-1) could be probably the best choice for patients with severe FXD, who require regular prophylaxi

Amamentação ao seio, amamentação com leite de vaca e o diabetes mellitus tipo 1: examinando as evidências Breast- feeding, bottle- feeding and the type 1 diabetes mellitus: examining the evidences

A etiologia do diabetes mellitus tipo 1 (DM1) envolve tanto herança genética como a exposição a fatores ambientais. Evidências de estudos epidemiológicos e experimentais sugerem que a dieta pode ser importante na etiopatogenia dessa doença. Em 1984, Borch-Johnsen e col. sugeriram, com base nos resultados de um estudo caso-controle, que o leite materno seria um fator de proteção para o DM1; esse efeito se daria devido às propriedades anti-infecciosas desse tipo de leite, ou pelo fato de que a amamentação ao seio evitaria que as crianças pudessem ser precocemente expostas a outros agentes etiológicos contidos nos substitutos do leite materno. Esses mesmos achados foram poste-riormente encontrados em diversos estudos, mas o papel do leite materno no aparecimento do DM1 ainda permanece controverso. Em 1992, Karjalainen e col., ao compararem os soros de indivíduos com e sem DM1, observaram, entre os diabéticos, altas concentrações de anticorpos anti-albumina bovina. Os autores postularam a hipótese de que a albumina bovina poderia atuar como desencadeadora do processo destrutivo das células ß do pâncreas e, conseqüentemente, do diabetes. Resultados conflitantes foram observados nas publicações que se sucederam a essa. Neste artigo, resumem-se e discutem-se os achados de diferentes pesquisadores que investigaram a importância desses fatores dietéticos para o aparecimento do DM1.<br>The aetiology of type 1 diabetes mellitus (DM1) includes genetic heritage and environmental exposure. Evidence from animal and epidemiological studies suggests that some diet components may play a role in the aetiology of DM1. In 1984, Borch-Johnsen et al. suggested, based on a case-control study, that breast-feeding was a protective factor for DM1, probably due its anti-infectious properties or because breast-feeding delays exposure to other etiologic agents in the diet. Afterwards, the same results were found in several studies but the role of breast milk in the development of DM1, is still subject to controversy. In 1992, Karjalainen et al., compared the blood serum of subjects with and without DM1 and they observed a higher concentration of anti-bovine albumin antibodies among diabetic subjects. The authors suggested that bovine albumin could act as a trigger of the destructive process of the pancreas and, in this way, lead to diabetes; discordant results have been observed in the literature since then. In this paper, we summarise and discuss the results found in different studies on dietary factors and DM1