Genetic diversity of Indian garlic core germplasm using agro-biochemical traits and SRAP markers

The characterization of garlic germplasm improves its utility, despite the fact that garlic hasn't been used much in the past. Garlic has an untapped genetic pool of immense economic and medicinal value in India. Hence, using heuristic core collection approach, a core set of 46 accessions were selected from 625 Indian garlic accessions based on 13 quantitative and five qualitative traits. The statistical measures (CV per cent, CR per cent, VR per cent) were used to sort the core set using Shannon-Wiener diversity index and the Nei diversity index. In addition, the variation within the core set was tested for 18 agro-morphological and six biochemical characteristics (allicin, phenol content, pyruvic acid, protein, allyl methyl thiosulfinate (AMTHS), and methyl allyl thiosulfinate (MATHS)). Further study of the core set's molecular diversity was performed using sequence related amplified polymorphism (SRAP) markers, which revealed a wide range of diversity among the core set's accessions, with an average polymorphism efficiency (PE) of 80.59 percent, polymorphism information content (PIC) of 0.29, effective multiplex ratio (EMR) of 3.51, and marker index (MI) of 0.99. The findings of this study will be useful in identifying high-yielding, elite garlic germplasm lines with the trait of interest. Since this core set is indicative of total germplasm, these selected breeding lines will be used for genetic improvement of garlic in the future.     

Clinical Validation of an Ultra High-Throughput Spiral Microfluidics for the Detection and Enrichment of Viable Circulating Tumor Cells

Background

Circulating tumor cells (CTCs) are cancer cells that can be isolated via liquid biopsy from blood and can be phenotypically and genetically characterized to provide critical information for guiding cancer treatment. Current analysis of CTCs is hindered by the throughput, selectivity and specificity of devices or assays used in CTC detection and isolation.

Methodology/Principal Findings

Here, we enriched and characterized putative CTCs from blood samples of patients with both advanced stage metastatic breast and lung cancers using a novel multiplexed spiral microfluidic chip. This system detected putative CTCs under high sensitivity (100%, n = 56) (Breast cancer samples: 12–1275 CTCs/ml; Lung cancer samples: 10–1535 CTCs/ml) rapidly from clinically relevant blood volumes (7.5 ml under 5 min). Blood samples were completely separated into plasma, CTCs and PBMCs components and each fraction were characterized with immunophenotyping (Pan-cytokeratin/CD45, CD44/CD24, EpCAM), fluorescence in-situ hybridization (FISH) (EML4-ALK) or targeted somatic mutation analysis. We used an ultra-sensitive mass spectrometry based system to highlight the presence of an EGFR-activating mutation in both isolated CTCs and plasma cell-free DNA (cf-DNA), and demonstrate concordance with the original tumor-biopsy samples.

Conclusions/Significance

We have clinically validated our multiplexed microfluidic chip for the ultra high-throughput, low-cost and label-free enrichment of CTCs. Retrieved cells were unlabeled and viable, enabling potential propagation and real-time downstream analysis using next generation sequencing (NGS) or proteomic analysis.     

Awareness and Attitude towards Human Papillomavirus (HPV) Vaccine among Medical Students in a Premier Medical School in India

Background

As preventing cancer with the help of a vaccine is a comparatively new concept, awareness and education about it will have important implication in the implementation of this strategy.

Materials and Methods

Present explorative questionnaire based survey included 618 MBBS students for final analysis.

Results

Majority of participants (89.6%) were well aware of the preventable nature of cervical cancer. Most of them (89.2%) knew that necessary factor responsible for cervical cancer is infection with high risk HPV. Awareness regarding the availability of vaccine against cervical cancer was 75.6%. Females had a better awareness regarding availability of vaccine, target population for vaccination and about the catch up program. Overall acceptance of HPV vaccine among the population studied was 67.8%. Medical teaching had a definitive impact on the understanding of this important public health issue. Females seemed to be more ready to accept the vaccine and recommend it to others. For our study population the most common source of information was medical school teaching. Majority of participants agreed that the most important obstacle in implementation of HPV vaccination program in our country is inadequate information and 86.2% wanted to be educated by experts in this regard.

Conclusion

HPV vaccine for primary prevention of cervical cancer is a relatively new concept. Health professional will be able to play a pivotal role in popularizing this strategy.     

“New”‐clear functions of PDK1: Beyond a master kinase?

Activation of cytosolic phosphoinositide-3 kinase (PI-3K) signaling pathway has been well established to regulate gene expression, cell cycle, and survival by feeding signals to the nucleus. In addition, strong evidences accumulated over the past few years indicate the presence of an autonomous inositol lipid metabolism and PI-3K signaling within the nucleus. Much less, however, is known about the role and regulation of this nuclear PI-3K pathway. Components of the PI-3K signaling pathway, including PI 3-kinase and its downstream kinase Akt, have been identified at the nuclear level. Consistent with the presence of a complete PI-3K signaling pathway in the nucleus, we have recently found that phosphoinositide-dependent kinase 1 (PDK1), a kinase functioning downstream of PI-3K and upstream of Akt, is a nucleo-cytoplasmic shuttling protein. In the present review, we update our current knowledge on the regulatory mechanisms and the functional roles of PDK1 nuclear translocation. We also summarize some of the kinase-independent activities of PDK1 in cell signaling.     

An efficient micropropagation protocol for Citrus jambhiri Lush. and assessment of clonal fidelity employing anatomical studies and RAPD markers

Citrus jambhiri (rough lemon) is considered a major rootstock source for a number of Citrus species. A simple method for micropropagation from nodal segments is reported. Nodal segments of C. jambhiri were inoculated on MS medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP), kinetin, and N6-(2-isopentenyl) adenosine (2iP). Maximum multiple shoot regeneration response (75?%) was observed with BAP at 3?mg?l^?1. Shoots were multiplied for 30?d on fresh medium with similar composition. A total of 67?% of the cultures showed multiplication with the optimum number of shoots (4.02) and height of shoots (1.81?cm) with BAP (3?mg?l^?1) alone. Maximum rooting response (87?%) was observed with naphthaleneacetic acid at 0.5?mg?l^?1. Transverse sections of shoot stems obtained in vivo (sampled from seedlings) and in vitro (regenerated from nodal segments), showed similar anatomies. Randomly amplified polymorphic DNA analysis confirmed that all the regenerated plants were genetically identical to their donor plant, suggesting absence of detectable genetic variation in the regenerated plantlets.     

Modulation of prion formation, aggregation, and toxicity by the actin cytoskeleton in yeast

Self-perpetuating protein aggregates transmit prion diseases in mammals and heritable traits in yeast. De novo prion formation can be induced by transient overproduction of the corresponding prion-forming protein or its prion domain. Here, we demonstrate that the yeast prion protein Sup35 interacts with various proteins of the actin cortical cytoskeleton that are involved in endocytosis. Sup35-derived aggregates, generated in the process of prion induction, are associated with the components of the endocytic/vacuolar pathway. Mutational alterations of the cortical actin cytoskeleton decrease aggregation of overproduced Sup35 and de novo prion induction and increase prion-related toxicity in yeast. Deletion of the gene coding for the actin assembly protein Sla2 is lethal in cells containing the prion isoforms of both Sup35 and Rnq1 proteins simultaneously. Our data are consistent with a model in which cytoskeletal structures provide a scaffold for generation of large aggregates, resembling mammalian aggresomes. These aggregates promote prion formation. Moreover, it appears that the actin cytoskeleton also plays a certain role in counteracting the toxicity of the overproduced potentially aggregating proteins.     

Cholinesterase inhibitory potential of different Alternaria spp. and their phylogenetic relationships

Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitors are used for the treatment of various disorders related to decline in acetylcholine levels in the brain by inhibiting the activity of the neurotransmitter AChE. The present study reports the potential of endophytic Alternaria spp. for their potential to produce cholinesterase inhibitors active against both acetylcholine and butyrylcholine. Twenty-nine isolates belonging to Alternaria spp. were isolated from different plants and screened. Variation with respect to inhibitor production was observed in different isolates. Out of 29 cultures screened, good cholinesterase (both AChE and BChE) inhibitory activity in range of 70–85% was observed in three isolates, whereas three showed only AChE inhibition. No correlation was observed in AChE and BChE inhibitor production. TLC bioautography for the inhibitor in the selected cultures evinced different Rf values of inhibitors indicating different nature of the compounds produced. In order to analyze evolutionary relationships between producer and non-producer strains, phylogenetic analysis of six producer and five non-producer strains was carried out using amplified ITS-I-5.8SrDNA-ITS-II region. Phylogenetic analysis revealed majority of the non-producer strains to be present on different clades indicating different evolutionary origins. The dual cholinesterase inhibitory activity and the diversity in the inhibitors produced by different isolates could prove to be novel sources of pharmaceutical as well as agriculturally important biomolecules after purification and characterization.     

Morphological differences between native and non-native pumpkinseed in traits associated with locomotion

Adaptive strategies in morphology can significantly influence the successful invasion and establishment of non-native species. Since its introduction, the pumpkinseed (Lepomis gibbosus), a sunfish of North American origin, has spread throughout most of Europe, including the Iberian Peninsula. We hypothesized that 12 morphological traits, functionally significant for locomotion, would differ according to geographic origin (native/non-native) and habitat type (fluvial/lacustrine). Using flow-through raceways, we simultaneously reared F₁ young-of-the-year pumpkinseed from two native and two non-native populations, produced from adults kept in a common environment. Morphometric measurements were recorded at the beginning and end of the 90-day rearing period. Median-fin size and placement differed significantly between native and non-native populations, whereas paired fin size differed between fluvial and lacustrine populations. Other functionally significant traits, such as body width, also differed between native and non-native populations. Spanish populations were considered to have acquired these adaptive external morphologies through successive generations, following the species’ range expansion through the variable environments of the Iberian Peninsula.     

Role of follicle stimulating hormone in the induction of hyperplasia during compensatory ovarian hypertrophy

In adult rats, removal of one ovary leads to an acute albeit transient rise in serum follicle stimulating hormone and an increase in the weight of the remaining ovary. In an attempt to correlate the high titre of endogenous follicle stimulating hormone with the changes taking place at the macromolecular level, the phenomenon of compensatory ovarian hypertrophy was studied for one cycle after hemiovariectomy at metoestrus in the adult, cycling female rats derived from the Holtzman strain. The significant finding with respect to hormonal changes was an acute follicle stimulating hormone surge commencing 6h post-unilateral ovariectomy, reaching a maximum at 12 h and declining thereafter, hitherto not reported in the Holtzman strain. Serum luteinizing hormone, prolactin, oestradiol-17β and testosterone remained unaltered while progesterone showed a decline at 6 h after surgery. There was an increase in the number of healthy class III (> 350 μm) follicles with a concomitant drop in atretic class III follicles 24 h post-unilateral ovariectomy. Analysis for DNA, RNA and protein content showed that all three constituents registered a continuous rise in the hypertrophying ovary up to 120h after surgery. When expressed as ?g/mg ovarian weight, the increase in DNA reached a maximum at 24 h and declined thereafter. The kinetics of DNA synthesis was followed by pulse labelling with [³H] thymidine at 18, 24, 36 and 48 h after unilateral ovariectomy. Maximum incorporation occurred at 36 h. Autoradiographic studies showed that the granulosa cells of healthy follicles preferentially incorporated the label. In an extension of this study, it was found that labelling index registered a significant increase following ovariectomy, the maximum being reached at 24 h especially in classIII follicles. The results clearly point out the crucial role of hyperplasia in the response of the contralateral ovary to the surgery and implicate the rise in follicle stimulating hormone as the primary signal for initiation of such a response. This raises the question whether in compensatory ovarian hypertrophy follicle stimulating hormone has a mitogenic role