A protease-based biosensor for the detection of schistosome cercariae

Parasitic diseases affect millions of people worldwide, causing debilitating illnesses and death. Rapid and cost-effective approaches to detect parasites are needed, especially in resource-limited settings. A common signature of parasitic diseases is the release of specific proteases by the parasites at multiple stages during their life cycles. To this end, we engineered several modular Escherichia coli and Bacillus subtilis whole-cell-based biosensors which incorporate an interchangeable protease recognition motif into their designs. Herein, we describe how several of our engineered biosensors have been applied to detect the presence and activity of elastase, an enzyme released by the cercarial larvae stage of Schistosoma mansoni. Collectively, S. mansoni and several other schistosomes are responsible for the infection of an estimated 200 million people worldwide. Since our biosensors are maintained in lyophilised cells, they could be applied for the detection of S. mansoni and other parasites in settings without reliable cold chain access

Meltrin β/ADAM19 Interacting with EphA4 in Developing Neural Cells Participates in Formation of the Neuromuscular Junction

BACKGROUND: Development of the neuromuscular junction (NMJ) is initiated by the formation of postsynaptic specializations in the central zones of muscles, followed by the arrival of motor nerve terminals opposite the postsynaptic regions. The post- and presynaptic components are then stabilized and modified to form mature synapses. Roles of ADAM (A Disintegrin And Metalloprotease) family proteins in the formation of the NMJ have not been reported previously. PRINCIPAL FINDINGS: We report here that Meltrin beta, ADAM19, participates in the formation of the NMJ. The zone of acetylcholine receptor alpha mRNA distribution was broader and excess sprouting of motor nerve terminals was more prominent in meltrin beta-deficient than in wild-type embryonic diaphragms. A microarray analysis revealed that the preferential distribution of ephrin-A5 mRNA in the synaptic region of muscles was aberrant in the meltrin beta-deficient muscles. Excess sprouting of motor nerve terminals was also found in ephrin-A5 knockout mice, which lead us to investigate a possible link between Meltrin beta and ephrin-A5-Eph signaling in the development of the NMJ. Meltrin beta and EphA4 interacted with each other in developing motor neurons, and both of these proteins localized in the NMJ. Coexpression of Meltrin beta and EphA4 strongly blocked vesicular internalization of ephrin-A5-EphA4 complexes without requiring the protease activity of Meltrin beta, suggesting a regulatory role of Meltrin beta in ephrin-A5-Eph signaling. CONCLUSION: Meltrin beta plays a regulatory role in formation of the NMJ. The endocytosis of ephrin-Eph complexes is required for efficient contact-dependent repulsion between ephrin and Eph. We propose that Meltrin beta stabilizes the interaction between ephrin-A5 and EphA4 by regulating endocytosis of the ephrinA5-EphA complex negatively, which would contribute to the fine-tuning of the NMJ during development

Pre-breeding blood urea nitrogen concentration and reproductive performance of Bonsmara heifers within different management systems

This study investigated the association between pre-breeding blood urea nitrogen (BUN) concentration and reproductive performance of beef heifers within different management systems in South Africa. Bonsmara heifers (n= 369) from five herds with different estimated levels of nitrogen intake during the month prior to the commencement of the breeding season were sampled in November and December 2010 to determine BUN concentrations. Body mass, age, body condition score (BCS) and reproductive tract score (RTS) were recorded at study enrolment. Trans-rectal ultrasound and/or palpation was performed 4–8 weeks after a 3- month breeding season to estimate the stage of pregnancy. Days to pregnancy (DTP) was defined as the number of days from the start of the breeding season until the estimated conception date. Logistic regression and Cox proportional hazards survival analysis were performed to estimate the association of pre-breeding BUN concentration with subsequent pregnancy and DTP, respectively. After stratifying for herd and adjusting for age, heifers with relatively higher prebreeding BUN concentration took longer to become pregnant when compared to those with relatively lower BUN concentration (P=0.011). In the herd with the highest estimated nitrogen intake (n=143), heifers with relatively higher BUN were less likely to become pregnant (P=0.013) and if they did, it was only later during the breeding season (P=0.017), after adjusting for body mass. These associations were not present in the herd (n=106) with the lowest estimated nitrogen intake (P>0.500). It is concluded that Bonsmara heifers with relatively higher pre-breeding BUN concentration, might be at a disadvantage because of this negative impact on reproductive performance, particularly when the production system includes high levels of nitrogen intake.Faculty of Veterinary Science, University of Pretoriahttp://link.springer.com/journal/112502015-08-31hb201