Functional Characterization of a Magnesium Transporter of Root Endophytic Fungus Piriformospora indica

Magnesium (Mg) is a crucial macronutrient required for the regular growth of plants. Here we report the identification, isolation and functional characterization of Mg-transporter PiMgT1 in root endophytic fungus Piriformospora indica. We also report the role of P. indica in the improvement of the Mg nutrition of the plant particularly under Mg deficiency condition. Protein BLAST (BLASTp) for conserved domains analysis showed that PiMgT1 belong to CorA like protein family of bacteria. We have also observed the presence of conserved ‘GMN’ signature sequence which suggests that PiMgT1 belongs to Mg transporter family. Phylogenetic analysis revealed that PiMgT1 clustered among fungal CorA family members nearer to basidiomycetes. Functionality of PiMgT1 was confirmed by complementation of a yeast magnesium transporter mutant CM66. We have observed that PiMgT1 restored the growth of mutant and showed comparable growth with that of WT. We found statistically significant (p < 0.05) two fold increase in the total intracellular Mg content of mutant complemented with PiMgT1 as compared to the mutant. These observations suggest that PiMgT1 is actively involved in Mg uptake by the fungus and may be helping in the nutritional status of the host plant

ART outcome in young women with premature ovarian aging

Background: Young women with signs of ovarian aging are a matter of concern as far as their reproductive performance is concerned. With more women approaching infertility centers with this problem, it becomes necessary to understand what reproductive outcomes are possible in such cases. Female age and basal Follicle stimulating hormone (FSH) level, both are strong independent predictors of In Vitro Fertilization (IVF) outcome. Objective: To correlate age-related basal FSH with IVF outcome in women with premature ovarian aging in gonadotropins-induced cycles. Materials and Methods: Between January 2011 and October 2012, a total of 135 women undergoing IVF and ICSI cycles with antagonist protocol were included in this retrospective cohort study. Basal FSH concentrations were measured and the women′s ages were calculated before they were undergoing pituitary desensitization and its correlation with assisted reproduction technique (ART) outcome was evaluated. Results: Increasing FSH was associated significantly with reduced number of oocytes retrieved, and embryos obtained. Young women with high FSH up to 20 produced less but good quality embryo′s resulting in sound pregnancy rate. Conclusion: FSH is a quantitative and age is a qualitative measure of ovarian reserve. Both are equally important in predicting IVF outcome. Basal FSH concentration should be restricted to counseling of patients on probability of achieving pregnancy, but should not be used to exclude them from fertility treatment

Sulfur nutrition and its role in plant growth and development

Sulfur is one of the essential nutrients that is required for the adequate growth and development of plants. Sulfur is a structural component of protein disulfide bonds, amino acids, vitamins, and cofactors. Most of the sulfur in soil is present in organic matter and hence not accessible to the plants. Anionic form of sulfur (SO42−) is the primary source of sulfur for plants that are generally present in minimal amounts in the soil. It is water-soluble, so readily leaches out of the soil. Sulfur and sulfur-containing compounds act as signaling molecules in stress management as well as normal metabolic processes. They also take part in crosstalk of complex signaling network as a mediator molecule. Plants uptake sulfate directly from the soil by using their dedicated sulfate transporters. In addition, plants also use the sulfur transporter of a symbiotically associated organism like bacteria and fungi to uptake sulfur from the soil especially under sulfur depleted conditions. So, sulfur is a very important component of plant metabolism and its analysis with different dimensions is highly required to improve the overall well-being of plants, and dependent animals as well as human beings. The deficiency of sulfur leads to stunted growth of plants and ultimately loss of yield. In this review, we have focused on sulfur nutrition, uptake, transport, and inter-organismic transfer to host plants. Given the strong potential for agricultural use of sulfur sources and their applications, we cover what is known about sulfur impact on the plant health. We identify opportunities to expand our understanding of how the application of soil microbes like AMF or other root endophytic fungi affects plant sulfur uptake and in turn plant growth and development

Functional characterization of a hexose transporter from root endophyte Piriformospora indica

Understanding the mechanism of photosynthate transfer at symbiotic interface by fungal monosaccharide transporter is of substantial importance. The carbohydrate uptake at the apoplast by the fungus is facilitated by PiHXT5 hexose transporter in root endophytic fungus Piriformospora indica. The putative PiHXT5 belongs to MFS superfamily with twelve predicted transmembrane helices. It possesses sugar transporter PFAM motif (PF0083) and MFS superfamily domain (PS50850). It contains the signature tags related to glucose transporter GLUT1 of human erythrocyte. PiHXT5 is regulated in response to mutualism as well as glucose concentration. We have functionally characterized PiHXT5 by complementation of hxt-null mutant of Saccharomyces cerevisiae EBY.VW4000. It is involved in transport of multiple sugars ranging from D-glucose, D-fructose, D-xylose, D-mannose, D-galactose with decreasing affinity. The uncoupling experiments indicate that it functions as H+/glucose co-transporter. Further, pH dependence analysis suggests that it functions maximum between pH 5 to 6. The expression of PiHXT5 is dependent on glucose concentration and was found to be expressed at low glucose levels (1 mM) which indicate its role as a high affinity glucose transporter. Our study on this sugar transporter will help in better understanding of carbon metabolism and flow in this agro-friendly fungus

Are women satisfied when using levonorgestrel-releasing intrauterine system for treatment of abnormal uterine bleeding?

Aim: To determine the efficacy of levonorgestrel intrauterine system (LNG IUS) in treatment of abnormal uterine bleeding (AUB) in women over 35 years and to determine satisfaction of users of LNG IUS in case of AUB. Materials and Methods: This was a multicentric, retrospective, and observational study. Case records of patients with AUB from the hospitals in Pune, Delhi, and Gurgaon for the last 6 years were examined. Records of 80 women who had an LNG IUS inserted were included. The chief complaints and their duration were recorded. Investigation results, histopathology reports, and date of LNG IUS insertion were noted. The incidence of spotting, heavy menstrual bleeding, pain, expulsion, and amenorrhea were recorded at 3, 6, 12, and 18 months following treatment. Following this a telephonic interview was conducted to determine the efficacy of LNG IUS in treating the symptoms. Patients′ satisfaction in percentage was also noted and they were asked if they would recommend the LNG IUS to other women. Results: The mean age of women was 42.3 years. 77.5% of the women had menorrhagia as the chief complaint, and the mean duration was 12 months. Fibroids and adenomyosis were the most common pathology, present in 21.3% and 20% of the patients respectively. At 3 months, spotting seemed to be the predominant symptom (59.4%) and only 15% women had heavy bleeding. 49.3% of women were asymptomatic at 6 months. 27.5% had amenorrhea by the end of 18 months. 14 women in whom the device was expelled or removed due to persistent symptoms, underwent hysterectomy at various stages during the study period. Overall patient satisfaction was high at about 80%. Furthermore, 73.8% patients agreed to recommend it to other women. Conclusion: LNG IUS seems to be a viable and effective treatment option for AUB in women after 35 years. There is a high rate of patient satisfaction in appropriately selected patients

Deciphering streptococcal biofilms

Streptococci are a diverse group of bacteria, which are mostly commensals but also cause a considerable proportion of life-threatening infections. They colonize many different host niches such as the oral cavity, the respiratory, gastrointestinal, and urogenital tract. While these host compartments impose different environmental conditions, many streptococci form biofilms on mucosal membranes facilitating their prolonged survival. In response to environmental conditions or stimuli, bacteria experience profound physiologic and metabolic changes during biofilm formation. While investigating bacterial cells under planktonic and biofilm conditions, various genes have been identified that are important for the initial step of biofilm formation. Expression patterns of these genes during the transition from planktonic to biofilm growth suggest a highly regulated and complex process. Biofilms as a bacterial survival strategy allow evasion of host immunity and protection against antibiotic therapy. However, the exact mechanisms by which biofilm-associated bacteria cause disease are poorly understood. Therefore, advanced molecular techniques are employed to identify gene(s) or protein(s) as targets for the development of antibiofilm therapeutic approaches. We review our current understanding of biofilm formation in different streptococci and how biofilm production may alter virulence-associated characteristics of these species. In addition, we have summarized the role of surface proteins especially pili proteins in biofilm formation. This review will provide an overview of strategies which may be exploited for developing novel approaches against biofilm-related streptococcal infections

Cross-serotype protection against group A Streptococcal infections induced by immunization with SPy_2191

A high number of serotypes makes vaccine development to group A Streptococcus (GAS) difficult. Here, the authors use a reverse vaccinology approach and identify SPy_2191 as conserved surface protein that inhibits GAS adhesion and invasion and induces cross-protective immunity in mice

Cross-serotype protection against group A Streptococcal infections induced by immunization with SPy_2191

Group A Streptococcus (GAS) infection causes a range of diseases, but vaccine development is hampered by the high number of serotypes. Here, using reverse vaccinology the authors identify SPy_2191 as a cross-protective vaccine candidate. From 18 initially identified surface proteins, only SPy_2191 is conserved, surface-exposed and inhibits both GAS adhesion and invasion. SPy_2191 immunization in mice generates bactericidal antibodies resulting in opsonophagocytic killing of prevalent and invasive GAS serotypes of different geographical regions, including M1 and M49 (India), M3.1 (Israel), M1 (UK) and M1 (USA). Resident splenocytes show higher interferon-γ and tumor necrosis factor-α secretion upon antigen re-stimulation, suggesting activation of cell-mediated immunity. SPy_2191 immunization significantly reduces streptococcal load in the organs and confers ~76-92% protection upon challenge with invasive GAS serotypes. Further, it significantly suppresses GAS pharyngeal colonization in mice mucosal infection model. Our findings suggest that SPy_2191 can act as a universal vaccine candidate against GAS infections