Immunological diagnosis as an adjunctive tool for an early diagnosis of tuberculous meningitis of an immune competent child in a low tuberculosis endemic country: A case report

Background: Pediatric tuberculous meningitis is a highly morbid, often fatal disease. Its prompt diagnosis and treat - ment saves lives, in fact delays in the initiation of therapy have been associated with high mortality rates. Case presentation: This is a case of an Italian child who was diagnosed with tuberculous meningitis after a history of a month of headache, fatigue and weight loss. Cerebrospinal fluid analysis revealed a lymphocytic pleocytosis with predominance and decreased glucose concentration. Microscopy and conventional diagnostic tests to identify Myco - bacterium tuberculosis were negative, while a non classical method based on intracellular cytokine flow cytometry response of CD4 cells in cerebral spinal fluid helped us to address the diagnosis, that was subsequently confirmed by a nested polymerase chain reaction amplifying a 123 base pair fragment of the M. tuberculosis DNA. Conclusions: We diagnosed tuberculous meningitis at an early stage through an innovative immunological approach, supported by a nested polymerase chain reaction for detection of M. tuberculosis DNA. An early diagnosis is required in order to promptly initiate a therapy and to increase the patient’s surviva

Rare splenic complications and specific serology: decisive diagnostic tools in two cases of visceral leishmaniasis

Introduction: Visceral leishmaniasis (VL) is a major endemic vector-borne disease in Southern Europe. We present two cases of VL, both characterized by splenic complications. Methods and results: Case 1: A 47-year-old female presented with effort angina, hepatosplenomegaly and pancytopenia. The clinical course was complicated by splenic infarction. Although bone marrow biopsy failed to show amastigotes, diagnosis was performed by a fast agglutinating screening test (FAST) and confirmed by a direct agglutinating test (DAT). The patient was treated successfully with AmBisome. Case 2: A 22-year-old male who had undergone a splenectomy to treat splenic rupture related to a minor trauma four months earlier presented with fever, nocturnal sweats and weight loss. The lack of pancytopenia was due to the absence of the spleen. The first biopsy did not identify parasites, but because the FAST had been positive, another bone marrow biopsy was performed, which demonstrated leishmaniasis. This patient was treated with the same schedule of AmBisome infusion. Discussion: 1) The clinical presentation of VL can be atypical, 2) splenic complications can characterize this disease, and 3) specific serology may be an important tool to reach a diagnosis

Conservação da Biodiversidade

Os conhecimentos gerados pela Ecologia da Paisagem têm sido ampla- mente utilizados para orientar ações práticas da Biologia da Conservação. A Ecologia da Paisagem tem contribuído em muitos avanços no conhecimento sobre o processo de perda e fragmentação de habitats (FAHRIG, 2003; HADDAD et al., 2015); os limiares ecológicos (RADFORD et al., 2005); a recuperação de áreas degradadas (TAMBOSI et al., 2014); a influência da matriz, corredores ecológicos (DAMSCHEN et al., 2006) e stepping stones na conectividade da paisagem (UEZU et al., 2008); os efeitos de borda (PFEI- FER et al., 2017); a percepção das espécies sobre a paisagem (UEZU et al.. 2005); e o impacto de diversos distúrbios antrópicos, tais como: a presença de estradas, mineração, barragens, entre outros. Esses conhecimentos têm implicações diretas sobre a priorização de áreas para criação de Unidades de Conservação, na seleção de áreas a serem recuperadas, no manejo de espécies exóticas e invasoras e na manutenção e conservação da diversidade genética, das espécies ameaçadas de extinção e dos serviços ecossistêmicos (LINDENMAYER et al., 2006). Essas correspondências criam uma forte interdependência entre Ecologia da Paisagem e Biologia da Conservação

Direct RNA Nanopore Sequencing of SARS-CoV-2 Extracted from Critical Material from Swabs

In consideration of the increasing prevalence of COVID-19 cases in several countries and the resulting demand for unbiased sequencing approaches, we performed a direct RNA sequencing (direct RNA seq.) experiment using critical oropharyngeal swab samples collected from Italian patients infected with SARS-CoV-2 from the Palermo region in Sicily. Here, we identified the sequences SARS-CoV-2 directly in RNA extracted from critical samples using the Oxford Nanopore MinION technology without prior cDNA retrotranscription. Using an appropriate bioinformatics pipeline, we could identify mutations in the nucleocapsid (N) gene, which have been reported previously in studies conducted in other countries. In conclusion, to the best of our knowledge, the technique used in this study has not been used for SARS-CoV-2 detection previously owing to the difficulties in the extraction of RNA of sufficient quantity and quality from routine oropharyngeal swabs. Despite these limitations, this approach provides the advantages of true native RNA sequencing and does not include amplification steps that could introduce systematic errors. This study can provide novel information relevant to the current strategies adopted in SARS-CoV-2 next-generation sequencing