Multisite Microprobes for Electrochemical Recordings in Biological Dynamics

For over 30 years, techniques have been developed that allow for the microscale (10-30 /mum) measurement of chemical signals with high temporal resolution (1-200 Hz). Such measurements, called in vivo electrochemical recordings, allow for the direct determination of neurotransmitter molecules and related compounds in biological systems. Multiple recordings, simultaneously performed at different, closely spaced, well defined locations throughout a three-dimensional tissue volume in the brain, are of interest in neuroscience. Developments in microelectronic techniques enable the fabrication of multi-electrode microprobes for recording extracellular action potentials generated by individual neurons simultaneously. A high-yield microfabrication process has been successfully developed for the fabrication of a novel semiconductor based, four-site silicon microprobe that involves a three-mask process and standard UV photolithography. A plasma process has been developed for dry etching of the gold electrodes and conducting lines. The electrochemical behavior of the microprobe is investigated by a high-speed computer-based in vitro electrochemical recording system. The electrochemical signals are measured at 5 Hz and varying gain. It is found that a selectivity of over 500:1 is achieved, and the signal to noise ratio of the recorded signal is particularly suitable for in vivo recordings

Multisite Microprobes for Electrochemical Recordings in Biological Dynamics

For over 30 years, techniques have been developed that allow for the microscale (10-30 /mum) measurement of chemical signals with high temporal resolution (1-200 Hz). Such measurements, called in vivo electrochemical recordings, allow for the direct determination of neurotransmitter molecules and related compounds in biological systems. Multiple recordings, simultaneously performed at different, closely spaced, well defined locations throughout a three-dimensional tissue volume in the brain, are of interest in neuroscience. Developments in microelectronic techniques enable the fabrication of multi-electrode microprobes for recording extracellular action potentials generated by individual neurons simultaneously. A high-yield microfabrication process has been successfully developed for the fabrication of a novel semiconductor based, four-site silicon microprobe that involves a three-mask process and standard UV photolithography. A plasma process has been developed for dry etching of the gold electrodes and conducting lines. The electrochemical behavior of the microprobe is investigated by a high-speed computer-based in vitro electrochemical recording system. The electrochemical signals are measured at 5 Hz and varying gain. It is found that a selectivity of over 500:1 is achieved, and the signal to noise ratio of the recorded signal is particularly suitable for in vivo recordings

Language in tuberculosis services: can we change to patient-centred terminology and stop the paradigm of blaming the patients?

The words 'defaulter', 'suspect' and 'control' have been part of the language of tuberculosis (TB) services for many decades, and they continue to be used in international guidelines and in published literature. From a patient perspective, it is our opinion that these terms are at best inappropriate, coercive and disempowering, and at worst they could be perceived as judgmental and criminalising, tending to place the blame of the disease or responsibility for adverse treatment outcomes on one side-that of the patients. In this article, which brings together a wide range of authors and institutions from Africa, Asia, Latin America, Europe and the Pacific, we discuss the use of the words 'defaulter', 'suspect' and 'control' and argue why it is detrimental to continue using them in the context of TB. We propose that 'defaulter' be replaced with 'person lost to follow-up'; that 'TB suspect' be replaced by 'person with presumptive TB' or 'person to be evaluated for TB'; and that the term 'control' be replaced with 'prevention and care' or simply deleted. These terms are non-judgmental and patient-centred. We appeal to the global Stop TB Partnership to lead discussions on this issue and to make concrete steps towards changing the current paradigm

A weather-based scheme to advise on limited chemical control of groundnut leaf spot diseases in India

Infection of groundnuts by pathogens causing early and late leaf spot diseases (Cercospora arachidicola [Mycosphaerella arachidis] and Phaeoisariopsis personata [M. berkeleyi], respectively) is strongly affected by accumulated daily leaf wetness periods and, in the rainy season, temperature is unlikely to severely limit infection. Earlier work relating patterns of leaf wetness to infection, was used to define a daily Wetness Index (WI) which was compared with infection on inoculated plants exposed in the crop for periods of 7 d. Infection was only severe when the 7-d WI total exceeded a value of 2.3. The proportion of leaves with one or more lesions on the main stem was used to assess the amount of inoculum in the crop. When the proportion of diseased leaves exceeded 10% and the WI total exceeded the threshold, application of a fungicide was advised. Successive sprays were separated by at least 14 d and a maximum of three sprays were applied in the growing season. Field trials showed that three sprays gave limited benefit where the disease pressure was severe, but substantial increases in pod and haulm yield were possible with only one or two fungicide applications in locations with less disease pressur

Assessing climate risks in rainfed farming using farmer experience, crop calendars and climate analysis

Climate risk assessment in cropping is generally undertaken in a top-down approach using climate records while critical farmer experience is often not accounted for. In the present study, set in south India, farmer experience of climate risk is integrated in a bottom-up participatory approach with climate data analysis. Crop calendars are used as a boundary object to identify and rank climate and weather risks faced by smallhold farmers. A semi-structured survey was conducted with experienced farmers whose income is predominantly from farming. Interviews were based on a crop calendar to indicate the timing of key weather and climate risks. The simple definition of risk as consequence × likelihood was used to establish the impact on yield as consequence and chance of occurrence in a 10-year period as likelihood. Farmers’ risk experience matches well with climate records and risk analysis. Farmers’ rankings of ‘good’ and ‘poor’ seasons also matched up well with their independently reported yield data. On average, a ‘good’ season yield was 1·5–1·65 times higher than a ‘poor’ season. The main risks for paddy rice were excess rains at harvesting and flowering and deficit rains at transplanting. For cotton, farmers identified excess rain at harvest, delayed rains at sowing and excess rain at flowering stages as events that impacted crop yield and quality. The risk assessment elicited from farmers complements climate analysis and provides some indication of thresholds for studies on climate change and seasonal forecasts. The methods and analysis presented in the present study provide an experiential bottom-up perspective and a methodology on farming in a risky rainfed climate. The methods developed in the present study provide a model for end-user engagement by meteorological agencies that strive to better target their climate information delivery

Interaction between SNAI2 and MYOD enhances oncogenesis and suppresses differentiation in Fusion Negative Rhabdomyosarcoma

Rhabdomyosarcoma (RMS) is an aggressive pediatric malignancy of the muscle, that includes Fusion Positive (FP)-RMS harboring PAX3/7-FOXO1 and Fusion Negative (FN)-RMS commonly with RAS pathway mutations. RMS express myogenic master transcription factors MYOD and MYOG yet are unable to terminally differentiate. Here, we report that SNAI2 is highly expressed in FN-RMS, is oncogenic, blocks myogenic differentiation, and promotes growth. MYOD activates SNAI2 transcription via super enhancers with striped 3D contact architecture. Genome wide chromatin binding analysis demonstrates that SNAI2 preferentially binds enhancer elements and competes with MYOD at a subset of myogenic enhancers required for terminal differentiation. SNAI2 also suppresses expression of a muscle differentiation program modulated by MYOG, MEF2, and CDKN1A. Further, RAS/MEK-signaling modulates SNAI2 levels and binding to chromatin, suggesting that the differentiation blockade by oncogenic RAS is mediated in part by SNAI2. Thus, an interplay between SNAI2, MYOD, and RAS prevents myogenic differentiation and promotes tumorigenesis

Mitochondrial Associated Ubiquitin Fold Modifier-1 Mediated Protein Conjugation in Leishmania donovani

In this report, we demonstrate the existence of the ubiquitin fold modifier-1 (Ufm1) and its conjugation pathway in trypanosomatid parasite Leishmania donovani. LdUfm1 is activated by E1-like enzyme LdUba5. LdUfc1 (E2) specifically interacted with LdUfm1 and LdUba5 to conjugate LdUfm1 to proteinaceous targets. Mass spectrometry analysis revealed that LdUfm1 is conjugated to Leishmania protein targets that are associated with mitochondria. Immunofluorescence experiments showed that Leishmania Ufm1, Uba5 and Ufc1 are associated with the mitochondria. The demonstration that all the components of this system as well as the substrates are associated with mitochondrion suggests it may have physiological roles not yet described in any other organism. Overexpression of a non-conjugatable form of LdUfm1 and an active site mutant of LdUba5 resulted in reduced survival of Leishmania in the macrophage. Since mitochondrial activities are developmentally regulated in the life cycle of trypanosomatids, Ufm1 mediated modifications of mitochondrial proteins may be important in such regulation. Thus, Ufm1 conjugation pathway in Leishmania could be explored as a potential drug target in the control of Leishmaniasis

Enhancement of immune response of HBsAg loaded poly(L-lactic acid) microspheres against Hepatitis B through incorporation of alum and chitosan

Purpose: Poly (L-lactic acid) (PLA) microparticles encapsulating Hepatitis B surface antigen (HBsAg) with alum and chitosan were investigated for their potential as a vaccine delivery system. Methods: The microparticles, prepared using a water-in-oil-in-water (w/o/w) double emulsion solvent evaporation method with polyvinyl alcohol (PVA) or chitosan as the external phase stabilising agent showed a significant increase in the encapsulation efficiency of the antigen. Results: PLA-Alum and PLA-chitosan microparticles induced HBsAg serum specific IgG antibody responses significantly higher than PLA only microparticles and free antigen following subcutaneous administration. Chitosan not only imparted a positive charge to the surface of the microparticles but was also able to increase the serum specific IgG antibody responses significantly. Conclusions: The cytokine assays showed that the serum IgG antibody response induced is different according to the formulation, indicated by the differential levels of interleukin 4 (IL-4), interleukin 6 (IL-6) and interferon gamma (IFN-γ). The microparticles eliciting the highest IgG antibody response did not necessarily elicit the highest levels of the cytokines IL-4, IL-6 and IFN-γ

Common variants in CLDN2 and MORC4 genes confer disease susceptibility in patients with chronic pancreatitis

A recent Genome-wide Association Study (GWAS) identified association with variants in X-linked CLDN2 and MORC4 and PRSS1-PRSS2 loci with Chronic Pancreatitis (CP) in North American patients of European ancestry. We selected 9 variants from the reported GWAS and replicated the association with CP in Indian patients by genotyping 1807 unrelated Indians of Indo-European ethnicity, including 519 patients with CP and 1288 controls. The etiology of CP was idiopathic in 83.62% and alcoholic in 16.38% of 519 patients. Our study confirmed a significant association of 2 variants in CLDN2 gene (rs4409525—OR 1.71, P = 1.38 x 10-09; rs12008279—OR 1.56, P = 1.53 x 10-04) and 2 variants in MORC4 gene (rs12688220—OR 1.72, P = 9.20 x 10-09; rs6622126—OR 1.75, P = 4.04x10-05) in Indian patients with CP. We also found significant association at PRSS1-PRSS2 locus (OR 0.60; P = 9.92 x 10-06) and SAMD12-TNFRSF11B (OR 0.49, 95% CI [0.31–0.78], P = 0.0027). A variant in the gene MORC4 (rs12688220) showed significant interaction with alcohol (OR for homozygous and heterozygous risk allele -14.62 and 1.51 respectively, P = 0.0068) suggesting gene-environment interaction. A combined analysis of the genes CLDN2 and MORC4 based on an effective risk allele score revealed a higher percentage of individuals homozygous for the risk allele in CP cases with 5.09 fold enhanced risk in individuals with 7 or more effective risk alleles compared with individuals with 3 or less risk alleles (P = 1.88 x 10-14). Genetic variants in CLDN2 and MORC4 genes were associated with CP in Indian patients