STABILITY INDICATING RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS DETERMINATION OF VILDAGLIPTIN AND METFORMIN IN PHARMACEUTICAL DOSAGE FORM

Objective: The present study was aimed to develop a rapid, accurate, linear, sensitive and validate stability-indicating high performance liquid chromatographic [RP-HPLC] method for determination of vildagliptin and metformin in pharmaceutical dosage form.Methods: The chromatographic separation was performed on kromasil-C18 column [4.5 x 250 mm; 5 µm] using a mobile phase consisting of 0.05 mmol potassium dihydrogen phosphate buffer: acetonitrile [80:20 v/v], [pH adjusted to 3.5 using orthophosphoric acid]. The flow rate is 0.9 ml/min and the detection was carried out at 263 nm.Results: The chromatographic condition, the peak retention time of metformin and vildagliptin were found to be 2.215 min and 2.600 min respectively. Stress testing was performed in accordance with an international conference on harmonization [ICH] Q1A R2 guidelines. The method was validated as per ICH Q2 R1 guidelines. The calibration curve was found to be linear in the concentration range of 5-17.5 µg/ml and 50-175 µg/ml for vildagliptin and metformin. The limit of detection and quantification was found to be 0.0182 µg/ml and 0.0553 µg/ml for vildagliptin and 0.4451 µg/ml and 1.3490 µg/ml for metformin respectively.Conclusion: A new sensitive, simple and stability indicating reverse-phase high-performance liquid chromatography [RP-HPLC] method has been developed and validated for the determination of vildagliptin and metformin. The proposed method can be used for routine determination of vildagliptin and metformin

Regeneration potential of different explants during micropropagation of neem tree (Azadirachta indica A. Juss.)

Azadirachta indica A. Juss., (Neem), a prodigious multipurpose tree, has immense potential to benefit mankind and to protect the environment. In order to investigate the effects of three different explants for its regeneration potential, de embryonated cotyledon, immature zygotic embryo and nodal segments from a 30 year old neem plus tree were used. Half strength MS medium with benzyl amino purine (3 mg/L) and naphthalene acetic acid (0.5 mg/L) and casein hydrolysate (1 g/L) was effective in shoot bud sprouting from both nodes and cotyledons. Half strength MS medium fortified with TDZ (0.2 mg/L) was effective for induction of somatic embryogenesis from zygotic embryos. Shoot buds initiated from the cotyledons produced a maximum number of shoots per explants (4.33) which on further sub culturing induced maximum multiple shoots (15) on half strength MS medium fortified with BAP (1.5 mg/L), NAA (0.5 mg/L) and CH (400 g /L) and the nodal explants induced only 4-5 axillary shoots on further sub culturing. Even though immature zygotic embryos produced more number of somatic embryos per explant (24.97) within a short time (30-45 days), the plantlet conversion was poor (25.52 %). In vitro rooting was observed in half strength MS medium supplemented with IBA (2 mg/L). The regeneration potential of de embryonated cotyledons through a simple regeneration system may be beneficial for efficient mass propagation of selected plus trees of neem

Community level barriers for cervical cancer screening in marginalized population

Background: In India, cervical cancer is the second common cause of cancer deaths among women of reproductive age, with 469 million Indian women at risk. High risk human papillomavirus genotypes mainly 16 and 18 account of cervical cancer. The burden of cervical cancer can be reduced by regular screening of human papillomavirus (HPV). There is no specific national program for cervical cancer screening. Eligible women have limited knowledge of screening and also limited access to preventive screenings.Methods: The study was conducted in the slum areas of Ahmedabad city in Gujarat. 1088 women between 30-45 years of age were recruited in the study and 536 women consented to give cervical samples for DNA based HPV testing. We collected information regarding knowledge and practice for cervical cancer and HPV along with demographic data.Results: Lack of knowledge and practices around cervical cancer and screening among community women was found. There is a lack of awareness about the importance of preventive healthcare and near absence of evidence-based practices. Sociodemographic characteristics are important predictors of participation in the screening program.Conclusions: In the Indian context, HPV testing is a cost-effective option to prevent cervical cancer. The burden of cervical cancer is incredibly high. With increased ability to accurately detect, population level HPV testing would reduce the burden of cervical cancer and the ultimate cost per person would be minimal, due to the country’s large population. There is a need to develop policy to ensure participation of women in the HPV based cervical cancer screening programs

Analysis of algal cell elements collected from temple tanks, rock ponds and seashore in Kovalam and Mahabalipuram, Tamil Nadu, India

Four species of micro and macroalgae collected from Mahabalipuram and Kovalam were analyzed for their elemental composition with the help of Perkin Elmer Optima DV Inductively Couple Plasma – Optical Emission Spectroscopy. Several significant elements e.g., Calcium, Magnesium, Sodium, Potassium and Iron were detected quantitatively. Calcium, Potassium, Magnesium were found in large amounts (22.461 – 626.385 ppm), Iron and Sodium were present in small quantities (2.391 – 6.167 ppm). The average quantity of Potassium was found to be highest among these algae (626.385 ppm), followed by Calcium (65.253 ppm) and Magnesium (22.461 ppm) and low quantity of Sodium (6.167 ppm) and Iron (2.391 ppm)

GC-MS-based metabolomics analysis unravels the therapeutic potential of Neolamarckia cadamba fruit peel

Kadam (Neolamarckia cadamba (Roxb.) is an evergreen tropical tree widely grown in Asia, particularly in India. Neolamarckia cadamba commonly known as kadam, cadamba or burflower tree. The roots, leaves, barks, and fruits of N. cadamba possess medicinal properties and are commonly used in the pharmaceutical industry. Fruit peels are the main waste and may contain various biologically active compounds. However, no prior knowledge about the therapeutic compounds of the peel. The objective of the present study was to unveil therapeutic compounds from the peel by Gas Chromatography–Mass Spectrometry (GC-MS) based metabolomics analysis. Metabolites from the kadam fruit peel were isolated and derivatized using MSTFA, characterized by the GC-MS analysis. Raw spectral data were pre-processed, and peak identification was performed using SHIMADZU Postrun analyse software. The metabolites in N. cadamba fruit peel were identified by comparing the peaks with the mass spectral reference database NIST v20. The results showed that the peel of kadam fruit contains 149 metabolites, which were further categorized into 46 different metabolite classes, with 52 different metabolic pathways and 63 biological functions. The principal roles of the metabolites were identified by functional annotation and enrichment analysis. It revealed that metabolites were responsible for anti-inflammation, anti-oxidant, anti-microbial, and anti-cancer properties. In summary, the peel of kadam fruit also contains various therapeutic compounds like other cadamba parts (i.e., roots, leaves, barks, and fruits). Further, comparing the peel with other parts discloses the peel-specific metabolites. The results obtained in this study could be useful for the pharmaceutical industry

The trypanosome transcriptome is remodelled during differentiation but displays limited responsiveness within life stages.

BACKGROUND: Trypanosomatids utilise polycistronic transcription for production of the vast majority of protein-coding mRNAs, which operates in the absence of gene-specific promoters. Resolution of nascent transcripts by polyadenylation and trans-splicing, together with specific rates of mRNA turnover, serve to generate steady state transcript levels that can differ in abundance across several orders of magnitude and can be developmentally regulated. We used a targeted oligonucleotide microarray, representing the strongly developmentally-regulated T. brucei membrane trafficking system and approximately 10% of the Trypanosoma brucei genome, to investigate both between-stage, or differentiation-dependent, transcriptome changes and within-stage flexibility in response to various challenges. RESULTS: 6% of the gene cohort are developmentally regulated, including several small GTPases, SNAREs, vesicle coat factors and protein kinases both consistent with and extending previous data. Therefore substantial differentiation-dependent remodeling of the trypanosome transcriptome is associated with membrane transport. Both the microarray and qRT-PCR were then used to analyse transcriptome changes resulting from specific gene over-expression, knockdown, altered culture conditions and chemical stress. Firstly, manipulation of Rab5 expression results in co-ordinate changes to clathrin protein expression levels and endocytotic activity, but no detectable changes to steady-state mRNA levels, which indicates that the effect is mediated post-transcriptionally. Secondly, knockdown of clathrin or the variant surface glycoprotein failed to perturb transcription. Thirdly, exposure to dithiothreitol or tunicamycin revealed no evidence for a classical unfolded protein response, mediated in higher eukaryotes by transcriptional changes. Finally, altered serum levels invoked little transcriptome alteration beyond changes to expression of ESAG6/7, the transferrin receptor. CONCLUSION: While trypanosomes regulate mRNA abundance to effect the major changes accompanying differentiation, a given differentiated state appears transcriptionally inflexible. The implications of the absence of a transcriptome response in trypanosomes for both virulence and models of life cycle progression are discussed.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

Evidence that low endocytic activity is not directly responsible for human serum resistance in the insect form of African trypanosomes.

BACKGROUND: In Trypanosoma brucei, the African trypanosome, endocytosis is developmentally regulated and substantially more active in all known mammalian infective stages. In both mammalian and insect stages endocytic activity is likely required for nutrient acquisition, but in bloodstream forms increased endocytosis is involved in recycling the variant surface glycoprotein and removing host immune factors from the surface. However, a rationale for low endocytic activity in insect stages has not been explored. Here we asked if endocytic down-regulation in the procyclic form was associated with resistance to innate trypanolytic immune factors in the blood meal or tsetse fly midgut. FINDINGS: Using a well-characterized procyclic parasite with augmented endocytic flux mediated via TbRab5A overexpression, we found that insect stage parasites were able to grow both in the presence of trypanosome lytic factor (TLF) provided in human serum, and also in tsetse flies. Additionally, by placing blood stage parasites in restricted glucose medium, we observed that enlargement of the flagellar pocket, a key morphology associated with defective endocytosis, manifests in parallel with loss of cellular ATP levels. CONCLUSIONS: These observations suggest that a high rate of endocytosis per se is insufficient to render insect form parasites sensitive to TLF or tsetse-derived trypanocidal factors. However, the data do suggest that endocytosis is energetically burdensome, as endocytic activity is rapidly compromised on energy depletion in bloodstream stages. Hence an important aspect of endocytic modulation in the nutrient-poor tsetse midgut is likely energetic conservation.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

Characterization of Little millet (Panicum sumatrense) varieties using Morphological descriptors and SSR based DNA fingerprinting

Little millet varieties are generally distinguished by morphological descriptors which are being used for seed certification and DUS characterization [1]. But in practical terms, these key differentiation descriptors between varieties of little millet are very fewer and hence difficult to differentiate germplasm accessions. Germplasm registration in NBPGR needs DNA fingerprint to show the uniqueness of germplasm in comparison to existing varieties. DNA fingerprinting is a better option to identify unique markers to differentiate the varieties. Available genomic resources are scarce since little millet is still considered to be an orphan crop. Therefore markers from other cereal genomes such as maize, pearl millet and barnyard millet that are been utilized for DNA fingerprinting purpose with a clue of cereal synteny relationship. Twenty-one morphological descriptors studies revealed that the variety ATL 1 is different from the other varieties for more than 16 morphological characters studied. DNA fingerprinting is attempted in five genotypes of little millets such as BL6, ATL 1, TNPsu 176, Co (Samai) 4, Paiyur 2 using cereal SSR markers. Among the 25 maize SSR markers used two markers viz., phi213984 and phi295450 scored polymorphism by the amplicon size of 310bp and 600bp respectively. From the 25 Pearl millet SSR markers used only one SSR marker found polymorphic at 305bp allele size for ATL 1 and Hence, SSR based DNA fingerprinting helped to differentiate ATL1, the newly released high yielding variety from other genotypes of little millets which can be used for varietal identification purpose

Conservation of leguminous crops and their wild relatives in Tamil Nadu, India, 2011

Based on the Memorandum of Understanding between the National Institute of Agrobiological Sciences, Japan and the Tamil Nadu Agricultural University, India, a field survey was conducted in Tamil Nadu State, India from 2^ to 6^ February, 2011. As a result, 39 accessions of leguminous plants consist of the genus Vigna , were recorded and seed samples consisting of 29 cultivated and 10 wild accessions were collected. All the seed materials collected were deposited at Tamil Nadu Agricultural University, India. Cultivation of Vigna stipulacea by local farmers and selling their seeds by local seed dealer were confirmed. Genetic erosion of traditional pulse landraces is rapidly proceeding in Tamil Nadu mainly due to an increase in the area of cash crops.本報告は，独立行政法人農業生物資源研究所ジーンバンクとインド，タミルナドゥ農業大学の間で2007年4月に締結した協同研究協定（MOU）に基づいて行ったインド，タミルナドゥ州における第三回目のマメ科植物遺伝資源の調査報告である．調査は，2011年2月2日～ 2月6日にかけて行った．調査の結果，ササゲ（Vigna）属栽培種5種29系統，ササゲ属野生種2種10系統を収集し，タミルナドゥ農業大学に保存した．タミルナドゥ州における豆類の価格は上がっており，儲かる作物になってきた．しかし，収穫作業に必要な労働者を雇用することが困難になってきているため，栽培をあきらめざるを得ない地主が多かった．今回の調査で，少々栽培化が進んで大型化したと考えられるマメ科植物Vigna stipulacea の農家による栽培が確認できた．V.stipulacea を栽培している農民は，1月から2月に水田にこの種の種子を散播し，緑肥，飼料，食料として利用している．種子は農民が自家採種し次年度に使うほか，町の種子屋でも販売していることを確認した．種子屋での聞き取りによれば，昨年は1500㎏の種子を販売し，販売価格はリョクトウ種子と同じで，1㎏あたり60ルピー（120円）であった．聞き取りを行ったすべての農民が，Sambal，Chatney，Dosai，Idli 等の伝統食品を作る場合，リョクトウやケツルアズキよりV.stipulacea の種子で作る方が風味があり，美味であるという感想を持っていた．また，栽培に関しても，リョクトウやケツルアズキに比べて病害虫に強く，V.stipulacea を栽培する際には一切農薬を散布する必要がないとのことであった