A Novel Antigen, Otubain Cysteine Peptidase of Leishmania donovani, for the Serodiagnosis of Visceral Leishmaniasis and for Monitoring Treatment Response.

Tests for visceral leishmaniasis (VL) are not uniformly effective for all endemic regions. In a serological assay, a novel antigen, otubain cysteine peptidase, compared with rK39, showed comparable sensitivity with Indian VL serum samples and prominently increased sensitivity with Brazilian samples, as well as improved monitoring of the treatment response.

Development and Clinical Evaluation of Serum and Urine-Based Lateral Flow Tests for Diagnosis of Human Visceral Leishmaniasis.

Visceral leishmaniasis (VL), a fatal parasitic infection, is categorized as being neglected among tropical diseases. The use of conventional tissue aspiration for diagnosis is not possible in every setting. The immunochromatography-based lateral flow assay (LFA) has attracted attention for a long time due to its ability to give results within a few minutes, mainly in resource-poor settings. In the present study, we optimized and developed the LFA to detect anti-Leishmania antibodies for VL diagnosis. The performance of the developed test was evaluated with serum and urine samples of Indian VL patients and Brazilian sera. The new test exploits well-studied and highly-sensitive purified antigens, LAg isolated from Leishmania donovani promastigotes and protein G conjugated colloidal-gold as a signal reporter. The intensity of the bands depicting the antigen-antibody complex was optimized under different experimental conditions and quantitatively analyzed by the ImageJ software. For the diagnosis of human VL in India, LFA was found to be 96.49% sensitive and 95% specific with serum, and 95.12% sensitive and 96.36% specific with urine samples, respectively. The sensitivity and specificity of LFA were 88.57% and 94.73%, respectively, for the diagnosis of Brazilian VL using patients' sera infected with Leishmania infantum. LFA is rapid and simple to apply, suitable for field usage where results can be interpreted visually and particularly sensitive and specific in the diagnosis of human VL. Serum and urine LFA may improve diagnostic outcomes and could be an alternative for VL diagnosis in settings where tissue aspiration is difficult to perform.

Is lactate production related to muscular fatigue? A pedagogical proposition using empirical facts.

The cause-effect relationship between lactic acid, acidosis, and muscle fatigue has been established in the literature. However, current experiments contradict this premise. Here, we describe an experiment developed by first-year university students planned to answer the following questions: 1) Which metabolic pathways of energy metabolism are responsible for meeting the high ATP demand during high-intensity intermittent exercise? 2) Which metabolic pathways are active during the pause, and how do they influence phosphocreatine synthesis? and 3) Is lactate production related to muscular fatigue? Along with these questions, students received a list of materials available for the experiment. In the classroom, they proposed two protocols of eight 30-m sprints at maximum speed, one protocol with pauses of 120 s and the other protocol with pauses of 20 s between sprints. Their performances were analyzed through the velocity registered by photocells. Blood lactate was analyzed before the first sprint and after the eighth sprint. Blood uric acid was analyzed before exercise and 15 and 60 min after exercises. When discussing the data, students concluded that phosphocreatine restoration is time dependent, and this fact influenced the steady level of performance in the protocol with pauses of 120 s compared with the performance decrease noted in the protocol with pauses of 20 s. As the blood lactate levels showed similar absolute increases after both exercises, the students concluded that lactate production is not related to the performance decrement. This activity allows students to integrate the understanding of muscular energy pathways and to reconsider a controversial concept with facts that challenge the universality of the hypothesis relating lactate production to muscular fatigue.