Pharmacogenetics of selective serotonin reuptake inhibitors (SSRI): A serotonin reuptake transporter (SERT)-based approach.

Neuropsychiatric disorders are considered to be the most common cause of disability worldwide. Serotonin and its transporter is a prominent paradigm in mood disorders. Response to selective serotonin reuptake inhibitors (SSRI) is altered due to heterogeneity in the serotonin transporter gene, SLC6A4 (solute carrier family 6 member 4). The reported polymorphisms are found to be in different regions of the transporter gene: promoter region (5-HTTLPR and various single nucleotide polymorphisms within it), intron (STin2), and exon 9 (I425V). The long and short alleles of the 5-HTTLPR gene, which are prevalent among variations, may mediate differential effects. In long allelic variant carriers, an increased response to SSRI and timely recovery is due to increased availability of SERT. Whereas, SERT availability is significantly decreased in short allelic carriers, necessitating a reduction in SSRI dosage due to the increased risk of adverse drug reactions. Thus, pharmacogenetic investigations are required to understand the impact of functional variations on the efficacy and tolerability of SSRI. Identifying the carrier variants may aid in clear-decision making of the treatment regimen, aiding the approach of personalized medication.

Safety aspects, probiotic potentials of yeast and lactobacillus isolated from fermented foods in North-Eastern India, and its anti-inflammatory activity.

Lactobacillus and yeast obtained from fermented foods in North-East India were tested for safety and probiotic properties. All the lactobacilli and yeast tested negative for the catalase, indole, urease, phenylalanine, hemolysis, gelatin hydrolysis, and biogenic amine production tests, indicating that they are safe to use as probiotics in food supplements. Lactiplantibacillus plantarum KGL3A (accession no. MG722814) was capable of resisting the replicated gastric fluid (pH 2) till 2 h of exposure, whereas both KGL3A and Lacticaseibacillus rhamnosus K4E (accession no. KX950834.1) strains were able to resist pH 3 till 2 h of exposure with a reduction in overall viable cell count from 7.48 log CFU/mL to 1.09 log CFU/mL and 7.77 log CFU/mL to 0.83 log CFU/mL, respectively. In vitro gastric juice simulation conditions were tolerated by the yeast Saccharomyces cerevisiae WBS2A. The cell surface hydrophobicity (CSH) towards hydrocarbons (n-hexadecane) was seen highest in L. plantarum KGL3A (77.16± 0.84%) and Limosilactobacillus fermentum KGL4 accession no. MF951099 (72.60 ± 2.33%). The percentage auto-aggregation ranged from 8.70 to 25.53 after 2 h, which significantly increased to 10.50 to 26.94 during the fifth hour for cultures. Also, a higher percentage of co-aggregation was found for the culture L. rhamnosus K4E with S. typhi (34.18 ± 0.03%), E. coli (32.97 ± 0.02 %) and S. aureus (26.33 ± 0.06 %) and for the yeast S. cerevisiae WBS2A, a higher percentage of co-aggregation was found with Listeria monocytogenes (25.77 ± 0.22%). The antioxidant activity and proteolytic activity were found to be higher for Lactobacillus helveticus K14 and L. rhamnosus K4E. The proportion of decreased cholesterol was noticeably higher in KGL4 (29.65 ± 4.30%). ß glucosidase activity was significantly higher in the L. fermentum KGL4 strain (0.359 ± 0.002), and α galactosidase activity was significantly higher in the L. rhamnosus K4E strain (0.415 ± 0.016). MTT assays suggested that KGL4 and WBS2A at a lower dose did not exhibit cytotoxicity.

Analysis of the Relationship between Scintillation Parameters, Multipath and ROTI.

Global Navigation Satellite System (GNSS) operation can be affected by several environmental factors, of which ionospheric scintillation is one of the most significant. Scintillation is usually characterized by two indices, namely the amplitude scintillation index (S4) and phase scintillation index (σφ). However, these two indices can only be generated by specialized GNSS receivers, which are not widely available all around the world. To popularize the study of scintillation, this article proposes to use more accessible parameters, namely multipath (MP) and rate of change of total electron content index (ROTI), to characterize scintillation. Using GPS data obtained on six days in total from three stations, namely PRU2 and SAO0P located in Sao Paulo, Brazil and SNA0P located in Antarctica, respectively, both the time series plots and 2D maps were generated to investigate the relationship of scintillation indices (S4 and σφ) with MP and ROTI. To prevent the effect of the real multipath error, a 30-degree satellite elevation mask is applied to all the data. As the scintillation indices S4 and σφ have a sampling interval of 1 min, MP and ROTI are calculated with the same sampling interval for a more direct comparison. The results show that the structural similarity (SSIM) and correlation coefficient (CC) between parameters was greater than 0.7 for 70% of outputs. In addition, the variogram and cross-variogram are applied to investigate the spatial structure of the MP, ROTI, S4 and σφ in order to support the results of SSIM and CC. With outputs in three forms, promising spatial and temporal relationships between parameters was observed.

Naturally occurring and tumor-associated variants of RNF167 promote lysosomal exocytosis and plasma membrane resealing.

Lysosomal exocytosis and resealing of damaged plasma membrane are essential for cellular homeostasis and tumor invasion. However, very little is known of the molecular machinery that regulates these physiological processes. Moreover, no mutations in any of the known regulators of lysosomal exocytosis in primary tumors of patients have been characterized. Here we demonstrate that RNF167-a, a lysosomal-associated ubiquitin ligase, negatively regulates lysosomal exocytosis by inducing perinuclear clustering of lysosomes. Importantly, we also characterized a set of novel natural mutations in RNF167-a, which are commonly found in diverse tumor types. We found that RNF167-a-K97N mutant, unlike the wild type, localizes in the cytoplasm and does not promote perinuclear lysosomal clustering. Furthermore, cells expressing RNF167-a-K97N exhibit dispersed lysosomes, increased exocytosis and enhanced plasma membrane repair. Interestingly, these functional features of RNF167-a-K97N were shared with a naturally occurring short version of RNF167 (isoform RNF167-b). In brief, the results presented here reveal a novel role of RNF167-a, as well as its natural variants RNF167-a-K97N and RNF167-b, as an upstream regulator of lysosomal exocytosis and plasma membrane resealing.

The ubiquitin ligase FBXW7 targets the centriolar assembly protein HsSAS-6 for degradation and thereby regulates centriole duplication.

Formation of a single new centriole from a pre-existing centriole is strictly controlled to maintain correct centrosome number and spindle polarity in cells. However, the mechanisms that govern this process are incompletely understood. Here, using several human cell lines, immunofluorescence and structured illumination microscopy methods, and ubiquitination assays, we show that the E3 ubiquitin ligase F-box and WD repeat domain-containing 7 (FBXW7), a subunit of the SCF ubiquitin ligase, down-regulates spindle assembly 6 homolog (HsSAS-6), a key protein required for procentriole cartwheel assembly, and thereby regulates centriole duplication. We found that FBXW7 abrogation stabilizes HsSAS-6 and increases its recruitment to the mother centriole at multiple sites, leading to supernumerary centrioles. Ultrastructural analyses revealed that FBXW7 is broadly localized on the mother centriole and that its presence is reduced at the site where the HsSAS-6-containing procentriole is formed. This observation suggested that FBXW7 restricts procentriole assembly to a specific site to generate a single new centriole. In contrast, during HsSAS-6 overexpression, FBXW7 strongly associated with HsSAS-6 at the centriole. We also found that SCFFBXW7 interacts with HsSAS-6 and targets it for ubiquitin-mediated degradation. Further, we identified putative phosphodegron sites in HsSAS-6, whose substitutions rendered it insensitive to FBXW7-mediated degradation and control of centriole number. In summary, SCFFBXW7 targets HsSAS-6 for degradation and thereby controls centriole biogenesis by restraining HsSAS-6 recruitment to the mother centriole, a molecular mechanism that controls supernumerary centrioles/centrosomes and the maintenance of bipolar spindles.

Effect of Probiotic Dietary Intervention on Calcium and Haematological Parameters in Geriatrics.

INTRODUCTION: Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. Sufficient calcium intake has been reported to support bone growth and prevent bone loss during the ageing process. AIM: To determine the effect of Lactobacillus helveticus MTCC 5463 probiotic dietary intervention on serum calcium & haematological parameters in geriatric population. MATERIALS AND METHODS: Healthy volunteers with age ranging from 64-74 years were recruited from the nearby residential areas in and around Anand, Gujarat. Study duration was from 2012 to 2015. Of the 112 subjects initially enrolled in the trial, 36 withdrew before the intervention because of not matching with criterias. Of the 76 participants, 5 subjects (4%) under test group and 12 subjects (11%) under placebo left the study. We had 59 subjects who successfully completed a double blind cross over trial. Probiotic fermented milk products (in form of "Lassi") was prepared by supplementing toned milk with honey and fermenting with probiotic Lactobacillus helveticus MTCC 5463 and Streptococcus thermophilus MTCC 5460. The final product had at least 10(8) CFU/ml of viable Lactobacillus helveticus MTCC 5463 at the time of feeding. During feeding period, 200 ml of fermented product containing the test strain to one group and a similar product but without the test strain as placebo were fed regularly at the time of breakfast in morning for 4 weeks. Subjects of each group were given a washout period of 4 weeks before they were crossed over and included to the other group. The study was approved by institutional ethics committee. RESULTS: The socio-demographic and clinical profiles were similar at baseline. The mean (SD) calcium level improved significantly in test {9.36 (0.45) vs 8.45 (0.61), p<0.001}. No significant effect was observed with respect to haemoglobin & haematological parameters. CONCLUSION: The well-documented probiotic Lactobacillus helveticus MTCC 5463 confirmed increase in serum calcium level but no effect on haematological parameters when administered to geriatrics.

Geriatric Respondents and Non-Respondents to Probiotic Intervention Can be Differentiated by Inherent Gut Microbiome Composition.

SCOPE: Probiotic interventions are known to have been shown to influence the composition of the intestinal microbiota in geriatrics. The growing concern is the apparent variation in response to identical strain dosage among human volunteers. One factor that governs this variation is the host gut microbiome. In this study, we attempted to define a core gut metagenome, which could act as a predisposition signature marker of inherent bacterial community that can help predict the success of a probiotic intervention. METHODS AND RESULTS: To characterize the geriatric gut microbiome, we designed primers targeting the 16S rRNA hypervariable region V2-V3 followed by semiconductor sequencing using Ion Torrent PGM. Among respondents and non-respondents, the chief genera of phylum Firmicutes that showed significant differences are Lactobacillus, Clostridium, Eubacterium, and Blautia (q < 0.002), while in the genera of phylum Proteobacteria included Shigella, Escherichia, Burkholderia and Camphylobacter (q < 0.002). CONCLUSION: We have identified potential microbial biomarkers and taxonomic patterns that correlate with a positive response to probiotic intervention in geriatric volunteers. Future work with larger cohorts of geriatrics with diverse dietary influences could reveal the potential of the signature patterns of microbiota for personalized nutrition.

Antibacterial and bioactive alpha- and beta-chitin hydrogel/nanobioactive glass ceramic/nano silver composite scaffolds for periodontal regeneration.

Alveolar bone loss and bone defects are the commonly encountered periodontal problems. Large defects do not heal spontaneously and thus require surgical interventions with bone substitutes. Bone grafts have the disadvantages of eliciting an immunologic response with subsequent graft rejection. The success rate of Guided Tissue Regeneration (GTR) is variable because of high susceptibility to infection. Thus emerged the important role of synthetic biomaterials and hence for this purpose we developed a nanocomposite scaffold, using alpha- and beta-chitin hydrogel with bioactive glass ceramic nanoparticles (nBGC) and silver nanoparticles (nAg) by lyophilization technique (aalpha and beta-chitin hydrogel/nBGC/nAg nanocomposite scaffold). The prepared nanoparticles and nanocomposite scaffolds were characterized. In addition, the porosity, swelling, mechanical properties, antibacterial activity, in vitro degradation and biomineralization, cell viability, cell attachment and cell proliferation ability of the prepared composite scaffolds were also evaluated. The results showed that alpha- and beta-chitin/nBGC/nAg composite scaffolds were porous and have the capacity to absorb fluids and swell. The composite scaffolds also showed enhanced antibacterial activity, bioactivity and controlled degradation in comparison to the control scaffolds. Cell viability studies proved the non-toxic nature of the nanocomposite scaffolds. Cell attachment and cell proliferation studies revealed the attachment and spreading nature of cells. All these studies revealed that, these antibacterial nanocomposite scaffolds could be a promising approach for the management of periodontal defects.

Probiotic Formulations: Application and Status as Pharmaceuticals-A Review.

The use of probiotics is a new way to control and treat infections in this modern era. Application of beneficial bacteria to protect against detrimental bacteria in the gastrointestinal tract and thus reap a positive health benefit is the basis of probiotic therapy. Probiotics have a long global history of traditional use. They are normally consumed through fermented foods and are currently sold mostly as ingredients in foods or nutritional supplements. They are also supplied as pharma products. Recent research has highlighted the probiotic potential in the treatment or prevention of disease conditions, maintenance of health, improving immunity and in the reduction in the risk of future diseases. But their position in the pharmaceutical industry is still not very clear. Clinical practitioners use probiotic pharma products mostly as supplements. Their status as drugs is still unclear. This review is aimed to analyze probiotics as pharmaceuticals, their current status as dietary supplements and drugs, existing probiotic preparations and future research needs.

Whole-genome shotgun sequencing of Lactobacillus rhamnosus MTCC 5462, a strain with probiotic potential.

Lactobacillus rhamnosus MTCC 5462 was isolated from infant gastrointestinal flora. The strain exhibited an ability to reduce cholesterol and stimulate immunity. The strain has exhibited positive results in alleviating gastrointestinal discomfort and good potential as a probiotic. We sequenced the whole genome of the strain and compared it to the published genome sequence of Lactobacillus rhamnosus GG (ATCC 53103).

Whole-genome shotgun sequencing of an Indian-origin Lactobacillus helveticus strain, MTCC 5463, with probiotic potential.

Lactobacillus helveticus MTCC 5463 was isolated from a vaginal swab from a healthy adult female. The strain exhibited potential probiotic properties, with their beneficial role in the gastrointestinal tract and their ability to reduce cholesterol and stimulate immunity. We sequenced the whole genome and compared it with the published genome sequence of Lactobacillus helveticus DPC4571.

Sodium alginate/poly(vinyl alcohol)/nano ZnO composite nanofibers for antibacterial wound dressings.

Sodium alginate (SA)/poly (vinyl alcohol) (PVA) fibrous mats were prepared by electrospinning technique. ZnO nanoparticles of size ∼160nm was synthesized and characterized by UV spectroscopy, dynamic light scattering (DLS), XRD and infrared spectroscopy (IR). SA/PVA electrospinning was further carried out with ZnO with different concentrations (0.5, 1, 2 and 5%) to get SA/PVA/ZnO composite nanofibers. The prepared composite nanofibers were characterized using FT-IR, XRD, TGA and SEM studies. Cytotoxicity studies performed to examine the cytocompatibility of bare and composite SA/PVA fibers indicate that those with 0.5 and 1% ZnO concentrations are less toxic where as those with higher concentrations of ZnO is toxic in nature. Cell adhesion potential of this mats were further proved by studying with L929 cells for different time intervals. Antibacterial activity of SA/PVA/ZnO mats were examined with two different bacteria strains; Staphylococcus aureus and Escherichia coli, and found that SA/PVA/ZnO mats shows antibacterial activity due to the presence of ZnO. Our results suggest that this could be an ideal biomaterial for wound dressing applications once the optimal concentration of ZnO which will give least toxicity while providing maximum antibacterial activity is identified.f.

Development of novel chitin/nanosilver composite scaffolds for wound dressing applications.

Antibiotic resistance of microorganisms is one of the major problems faced in the field of wound care and management resulting in complications like infection and delayed wound healing. Currently a lot of research is focused on developing newer antimicrobials to treat wounds infected with antibiotic resistant microorganisms. Silver has been used as an antimicrobial agent for a long time in the form of metallic silver and silver sulfadiazine ointments. Recently silver nanoparticles have come up as a potent antimicrobial agent and are finding diverse medical applications ranging from silver based dressings to silver coated medical devices. Chitin is a natural biopolymer with properties like biocompatibility and biodegradability. It is widely used as a scaffold for tissue engineering applications. In this work, we developed and characterized novel chitin/nanosilver composite scaffolds for wound healing applications. The antibacterial, blood clotting and cytotoxicity of the prepared composite scaffolds were also studied. These chitin/nanosilver composite scaffolds were found to be bactericidal against S. aureus and E. coli and good blood clotting ability. These results suggested that these chitin/nanosilver composite scaffolds could be used for wound healing applications.

A point mutation in a silencer module reduces the promoter activity for the human mercaptopyruvate sulfurtransferase.

A promoter region of human mercaptopyruvate sulfurtransferase (MST) [EC 2.8.1.2] is G+C-rich and TATA-less, showing features of a house-keeping gene. In the core promoter, a GC box (-284:GGGGCGTGGC:-275) and an initiator (-219:TTATATG:-225) are found. A cap site hunting analysis for human liver cDNA revealed four possible transcriptional start sites, nucleotides -223, -159, -35 and -25. Point mutagenesis and deletion studies suggest that a module of the silencer element is -394:GCTG:-391. A replacement of -391G to C lost the silencer function; on the other hand, a replacement of -394G to T or C, -393C to T or -392T to G markedly reduced the promoter activity.

New aspects in pathogenesis of konzo: neural cell damage directly caused by linamarin contained in cassava (Manihot esculenta Crantz).

Epidemic spastic paraparesis (konzo) found in tropical and subtropical countries is known to be caused by long-term intake of cassava (Manihot esculenta Crantz), which contains a cyanoglucoside linamarin (alpha-hydroxyisobutyronitrile-beta-d-glucopyranoside). It has been reported that linamarin is enzymatically converted to cyanide by bacteria in the intestine, and this is absorbed into the blood and then damages neural cells. However, unmetabolized linamarin was found in the urine after oral administration of cassava; thus, we hypothesized that konzo could be caused by direct toxicity of the unmetabolized linamarin that was transferred to the brain and could be transported into neural cells via a glucose transporter. In the present study it was confirmed that linamarin directly damaged neural culture pheochromocytoma cell (PC) 12 cells; 0.10 mm-linamarin caused cell death at 13.31 (SD 2.07) %, which was significantly different from that of control group (3.18 (SD 0.92) %, P=0.0004). Additional 10 microM-cytochalasin B, an inhibitor of a glucose transporter, prevented cell death: the percentage of dead cells significantly decreased to 6.06 (SD 1.98), P=0.0088). Furthermore, glucose also prevented cell death. These present results strongly suggest that linamarin competes with cytochalasin B and glucose for binding to a glucose transporter and enters into cells via glucose transporter.

Cassava diet--a cause for mucopolysaccharidosis?

Studies were carried out to determine the changes in glycosaminnoglycan (GAG) metabolism in rats fed cassava with varying cyanoglucoside levels and two levels of protein. Results indicated that there was an enhancement in the level of total and individual GAG with a corresponding reduction in the activity of enzymes involved in the degradation of glycosaminoglycan. These changes were significant for rats given a cassava diet (raw and boiled cassava) and low protein. The changes in total and individual GAG and the decrease in the activity of degrading enzymes was more for high cyanide (raw cassava) groups compared with other groups showing that consumption of untreated cassava is an additive factor for the promotion of mucopolysaccharidosis.

Hyperglycemic effect of low protein cassava diet.

Hyperglycemic effect of cassava diet in presence of varying amounts of protein has been carried out. The rats fed a low protein high cyanide diet showed an increase in the blood glucose and a decrease in the liver glycogen. The activity of glycogen phosphorylase, glucose 6-phosphatase and phosphoglucomutase showed higher levels in the liver of low protein high cyanide group compared to the control group. Also, the activity of hexokinase, and isocitrate dehydrogenase activity in the liver of high cyanide low protein were significantly low. The results suggests that cassava diet with the low protein can induce hyperglycemia.

Effect of protein supplemented cassava diet in rats.

The effect of feeding rats with cassava diet (tapioca) has been investigated with respect to the activities of superoxide dismutase, catalase and levels of TBARS. By varying the levels of protein in the diet and cyanide content of cassava, the possible role of a high protein diet in the prevention of oxidant stress has been shown. Rhodanese which detoxifes cyanide is also found to be inhibited in rats fed low protein diet while the inhibition was only marginal in the high protein fed groups.