Glutamate, aspartate and nucleotide transporters in the SLC17 family form four main phylogenetic clusters: evolution and tissue expression

<p>Abstract</p> <p>Background</p> <p>The SLC17 family of transporters transports the amino acids: glutamate and aspartate, and, as shown recently, also nucleotides. Vesicular glutamate transporters are found in distinct species, such as <it>C. elegans</it>, but the evolutionary origin of most of the genes in this family has been obscure.</p> <p>Results</p> <p>Our phylogenetic analysis shows that the SLC17 family consists of four main phylogenetic clades which were all present before the divergence of the insect lineage. One of these clades has not been previously described and it is not found in vertebrates. The clade containing Slc17a9 had the most restricted evolutionary history with only one member in most species. We detected expression of Slc17a1-17a4 only in the peripheral tissues but not in the CNS, while Slc17a5- Slc17a9 are highly expressed in both the CNS and periphery.</p> <p>Conclusions</p> <p>The <it>in situ </it>hybridization studies on vesicular nucleotide transporter revealed high expression throughout the cerebral cortex, certain areas in the hippocampus and in specific nuclei of the hypothalamus and thalamus. Some of the regions with high expression, such as the medial habenula and the dentate gyrus of the hippocampus, are important sites for purinergic neurotransmission. Noteworthy, other areas relying on purine-mediated signaling, such as the molecular layer of the dentate gyrus and the periaqueductal gray, lack or have a very low expression of Slc17a9, suggesting that there could be another nucleotide transporter in these regions.</p

ENDOSULFAN RESISTANCE PROFILE OF SOIL BACTERIA AND POTENTIAL APPLICATION OF RESISTANT STRAINS IN BIOREMEDIATION

Abstract. In the present study, bacterial strains were isolated from the soils of Wayanad District, Kerala, India and the isolates were tested for their tolerance to endosulfan and potential in bioremediation technology. Pesticide contamination in the soils, soil physico-chemical characteristics and socio-economic impacts of pesticide application were also analyzed. 28 pesticide compounds in the soil samples were analyzed and the results revealed that there was no pesticide residues in the soils. As per the survey conducted the pesticide application is very high in the study area and the level of awareness among the farmers was very poor regarding the method of application and its socio-economic and ecological impacts. A total of 9 bacterial strains were isolated with 50µg/ml of endosulfan in the isolating media and the results showed that most of the bacterial strains were highly resistance to endosulfan. Out of the 9 strains isolated 6 were highly resistant to endosulfan (500-700µg/ml) and the other 3 isolates showed the resistance of 250-500µg/ml. From the studied isolate, isolate 9 demonstrating prolific growth and high resistance was selected to check their capability to degrade endosulfan over time. Identification of the selected strain reveals that it belongs to the genus Bacillus. Results of endosulfan removal studies showed that with increase in time, the biomass of the bacterial strains increased. The complete disappearance of endosulfan from the spiked and inoculated broth during the first day of incubation (24 hour interval) was observed. While the control flask showed the presence of endosulfan during the experimental period. Pesticide resistant bacteria are widely distributed in the soils of selected study area and the tolerance varied between bacteria even though they were isolated 1 School of Environmental Sciences, Mahatma Gandhi University, Kottayam, Kerala, India. [email protected] 2 M S Swaminathan Research Foundation, Community Agrobiodiversity Centre, Kalpetta Wayanad, Kerala, India. Chandini P.K, Jaysooryan K.K, Rinoy Varghese, Sreedharan K, Smitha K. P. 128 from the soils of the same area. The selected Bacillus species carry the ability to degrade endosulfan at accelerated rates and it could be useful in framing a bioremediation strategy for pesticide contaminated soil and water environments

The obesity gene, TMEM18, is of ancient origin, found in majority of neuronal cells in all major brain regions and associated with obesity in severely obese children

<p>Abstract</p> <p>Background</p> <p>TMEM18 is a hypothalamic gene that has recently been linked to obesity and BMI in genome wide association studies. However, the functional properties of TMEM18 are obscure.</p> <p>Methods</p> <p>The evolutionary history of TMEM18 was inferred using phylogenetic and bioinformatic methods. The gene's expression profile was investigated with real-time PCR in a panel of rat and mouse tissues and with immunohistochemistry in the mouse brain. Also, gene expression changes were analyzed in three feeding-related mouse models: food deprivation, reward and diet-induced increase in body weight. Finally, we genotyped 502 severely obese and 527 healthy Swedish children for two SNPs near TMEM18 (rs6548238 and rs756131).</p> <p>Results</p> <p>TMEM18 was found to be remarkably conserved and present in species that diverged from the human lineage over 1500 million years ago. The TMEM18 gene was widely expressed and detected in the majority of cells in all major brain regions, but was more abundant in neurons than other cell types. We found no significant changes in the hypothalamic and brainstem expression in the feeding-related mouse models. There was a strong association for two SNPs (rs6548238 and rs756131) of the TMEM18 locus with an increased risk for obesity (p = 0.001 and p = 0.002).</p> <p>Conclusion</p> <p>We conclude that TMEM18 is involved in both adult and childhood obesity. It is one of the most conserved human obesity genes and it is found in the majority of all brain sites, including the hypothalamus and the brain stem, but it is not regulated in these regions in classical energy homeostatic models.</p

Functional Characterization of Centrally Expressed Solute Carriers and G Protein-Coupled Receptors

Transmembrane proteins are gatekeepers of the cells; controlling the transport of substrates as well as communicating signals among cells and between the organelles and cytosol. Solute carriers (SLC) and G protein-coupled receptors (GPCR) are the largest family of membrane transporters and membrane receptors respectively. The overall aim of this thesis was to provide a basic understanding of some of the novel SLCs and GPCRs with emphasis on expression, transport property, evolution and probable function. The first part of the thesis directs towards the study of some novel solute carriers. In an initial study, we provided an overall picture of the sequence relationship and tissue expression of 14 diverse atypical SLCs confirming some of their evolutionary conservation and highly specific expression pattern. The focus then was on the SLC17 family (mainly vesicular proteins) and a novel member named Slc17a9. This study revealed that SLC17 family could be divided into four main phylogenetic clades which were all present before the divergence of the insect lineage with Slc17a9 having the most restricted evolutionary history. Detailed expression study of Slc17a9 in the mouse brain suggests that it is also expressed in some regions important for purinergic neurotransmission. Further, we deorphanised an aminoacid transporter Slc38a7 which was expressed in a majority of neurons in the CNS and showed that it preferably mediate transport of L–glutamine and L–histidine. The second part of the thesis focuses on the study of two GPCRs belonging to the Rhodopsin superfamily, Gpr162 and Gpr153. A phylogenetic analysis revealed that both Gpr153 and Gpr162 originated from a common ancestor before the radiation of the mammalian lineage. Expression study revealed that Gpr162 had a predominant expression in the CNS and relatively lower expression in the other tissue tested whereas Gpr153 had a more widespread and similar expression pattern in both CNS and peripheral tissues. The functional studies of the two GPCRs were done using the antisense oligodeoxynucleotide knockdown rat model. These studies provided evidence linking the orphan Gpr162 gene with the regulation of food intake– related behaviour whereas Gpr153 gene caused only a slight reduction in food intake

Functional Characterization of Centrally Expressed Solute Carriers and G Protein-Coupled Receptors

Transmembrane proteins are gatekeepers of the cells; controlling the transport of substrates as well as communicating signals among cells and between the organelles and cytosol. Solute carriers (SLC) and G protein-coupled receptors (GPCR) are the largest family of membrane transporters and membrane receptors respectively. The overall aim of this thesis was to provide a basic understanding of some of the novel SLCs and GPCRs with emphasis on expression, transport property, evolution and probable function. The first part of the thesis directs towards the study of some novel solute carriers. In an initial study, we provided an overall picture of the sequence relationship and tissue expression of 14 diverse atypical SLCs confirming some of their evolutionary conservation and highly specific expression pattern. The focus then was on the SLC17 family (mainly vesicular proteins) and a novel member named Slc17a9. This study revealed that SLC17 family could be divided into four main phylogenetic clades which were all present before the divergence of the insect lineage with Slc17a9 having the most restricted evolutionary history. Detailed expression study of Slc17a9 in the mouse brain suggests that it is also expressed in some regions important for purinergic neurotransmission. Further, we deorphanised an aminoacid transporter Slc38a7 which was expressed in a majority of neurons in the CNS and showed that it preferably mediate transport of L–glutamine and L–histidine. The second part of the thesis focuses on the study of two GPCRs belonging to the Rhodopsin superfamily, Gpr162 and Gpr153. A phylogenetic analysis revealed that both Gpr153 and Gpr162 originated from a common ancestor before the radiation of the mammalian lineage. Expression study revealed that Gpr162 had a predominant expression in the CNS and relatively lower expression in the other tissue tested whereas Gpr153 had a more widespread and similar expression pattern in both CNS and peripheral tissues. The functional studies of the two GPCRs were done using the antisense oligodeoxynucleotide knockdown rat model. These studies provided evidence linking the orphan Gpr162 gene with the regulation of food intake– related behaviour whereas Gpr153 gene caused only a slight reduction in food intake

Functional Characterization of Centrally Expressed Solute Carriers and G Protein-Coupled Receptors

Transmembrane proteins are gatekeepers of the cells; controlling the transport of substrates as well as communicating signals among cells and between the organelles and cytosol. Solute carriers (SLC) and G protein-coupled receptors (GPCR) are the largest family of membrane transporters and membrane receptors respectively. The overall aim of this thesis was to provide a basic understanding of some of the novel SLCs and GPCRs with emphasis on expression, transport property, evolution and probable function. The first part of the thesis directs towards the study of some novel solute carriers. In an initial study, we provided an overall picture of the sequence relationship and tissue expression of 14 diverse atypical SLCs confirming some of their evolutionary conservation and highly specific expression pattern. The focus then was on the SLC17 family (mainly vesicular proteins) and a novel member named Slc17a9. This study revealed that SLC17 family could be divided into four main phylogenetic clades which were all present before the divergence of the insect lineage with Slc17a9 having the most restricted evolutionary history. Detailed expression study of Slc17a9 in the mouse brain suggests that it is also expressed in some regions important for purinergic neurotransmission. Further, we deorphanised an aminoacid transporter Slc38a7 which was expressed in a majority of neurons in the CNS and showed that it preferably mediate transport of L–glutamine and L–histidine. The second part of the thesis focuses on the study of two GPCRs belonging to the Rhodopsin superfamily, Gpr162 and Gpr153. A phylogenetic analysis revealed that both Gpr153 and Gpr162 originated from a common ancestor before the radiation of the mammalian lineage. Expression study revealed that Gpr162 had a predominant expression in the CNS and relatively lower expression in the other tissue tested whereas Gpr153 had a more widespread and similar expression pattern in both CNS and peripheral tissues. The functional studies of the two GPCRs were done using the antisense oligodeoxynucleotide knockdown rat model. These studies provided evidence linking the orphan Gpr162 gene with the regulation of food intake– related behaviour whereas Gpr153 gene caused only a slight reduction in food intake

Assessment of critical failure factors (CFFs) of lean Six Sigma in real life scenario : evidence from manufacturing and service industries

Purpose: Many projects focus on the reliable operation of the activities in the project. Any failure in the process activities leads to major problems resulting in waste, defects, equipment damage, which has a direct impact on the consumers. In addition, Lean Six Sigma (LSS) is not new to this issue. LSS projects have faced an interruption in the process flow and unforeseen defects. Therefore, the purpose of this paper is to identify the vital failure factors of LSS projects. Design/methodology/approach: Through extant literature review, the researchers found 44 critical failure factors (CFFs) of LSS. Using the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) SIMOS approach, the decision makers’ (DMs) rating and weight for each factor were collected. Moreover, the study was conducted in both the manufacturing and service industries to identify the impact of CFFs in LSS projects. Findings: CFFs and their evaluation have received little attention in the literature. Most of the previous studies deal only with the identification of the success factors in general. Therefore, the study identified 44 CFFs and ranked them through DMs. In addition, the TOPSIS SIMOS approach ranked the vital failure factors enabling the management to avert the LSS project from failures. Research limitations/implications: The study focused on project failures due to CFFs of LSS. Nevertheless, it did not consider other failure factors of project and knowledge management. Further, the sample used to test the approach was considerably small. Therefore, the study can be repeated with significant samples and the vital failure factors compared. Practical implications: In real-life application, all the parameters in the LSS project need to be understood in a better manner. In such a condition, the practitioner needs to know that the project never fails due to the CFFs and TOPSIS SIMOS approach can prevent the LSS project failures. Originality/value: The study applied TOPSIS SIMOS approach to rank the CFFs in an LSS project, which is first of its kind and aids the practitioners to make the right decisions in the business environment. © 2018, Emerald Publishing Limited

Endosulfan Resistance Profile of Soil Bacteria and Potential Application of Resistant Strains in Bioremediation

In the present study, bacterial strains were isolated from the soils of Wayanad District, Kerala, India and the isolates were tested for their tolerance to endosulfan and potential in bioremediation technology. Pesticide contamination in the soils, soil physico-chemical characteristics and socio-economic impacts of pesticide application were also analyzed. 28 pesticide compounds in the soil samples were analyzed and the results revealed that there was no pesticide residues in the soils. As per the survey conducted the pesticide application is very high in the study area and the level of awareness among the farmers was very poor regarding the method of application and its socio-economic and ecological impacts. A total of 9 bacterial strains were isolated with 50μg/ml of endosulfan in the isolating media and the results showed that most of the bacterial strains were highly resistance to endosulfan. Out of the 9 strains isolated 6 were highly resistant to endosulfan (500- 700μg/ml) and the other 3 isolates showed the resistance of 250-500μg/ml. From the studied isolate, isolate 9 demonstrating prolific growth and high resistance was selected to check their capability to degrade endosulfan over time. Identification of the selected strain reveals that it belongs to the genus Bacillus. Results of endosulfan removal studies showed that with increase in time, the biomass of the bacterial strains increased. The complete disappearance of endosulfan from the spiked and inoculated broth during the first day of incubation (24 hour interval) was observed. While the control flask showed the presence of endosulfan during the experimental period. Pesticide resistant bacteria are widely distributed in the soils of selected study area and the tolerance varied between bacteria even though they were isolated from the soils of the same area. The selected Bacillus species carry the ability to degrade endosulfan at accelerated rates and it could be useful in framing a bioremediation strategy for pesticide contaminated soil and water environment