Screening and Evaluation of in vivo and in vitro Generated Insulin Plant (Vernonia divergens) for Antimicrobial and Anticancer Activities

Vernonia divergens Benth., commonly known as “Insulin Plant” (Fam: Asteraceae) is a potent sugar killer. Locally the leaves of the plant, boiled in water are successfully administered to a large number of diabetic patients. The present study evaluates the putative anti-diabetic ingredients, isolated from the in vivo and in vitro grown plantlets of V. divergens for their antimicrobial and anticancer activities. Sterilized explants of nodal segments were cultured on MS (Musashige and Skoog, 1962) medium in presence of different combinations of hormones. Multiple shoots along with bunch of roots were regenerated at 1mg l-1 BAP and 0.5 mg l-1 NAA. Micro-plantlets were separated and sub-cultured on the double strength (2X) of the above combination of hormones leading to increased length of roots and shoots. These plantlets were successfully transferred to soil and survived well in nature. The ethanol extract of plantlets from both in vivo & in vitro sources were prepared in soxhlet extractor and then concentrated to dryness under reduced pressure in rotary evaporator. Thus obtainedconcentrated extracts showed significant inhibitory activity against gram negative bacteria like Escherichia coli and Pseudomonas aeruginosa but no inhibition was found against gram positive bacteria. Further, these ethanol extracts were screened for in vitro percentage cytotoxicity at different time periods (24 h, 48 h and 72 h) of different dilutions. The in vivo plant extract inhibited the growth of EAC mouse cell lines in the range of 65, 66, 78, and 88% at 100, 50, 25 & 12.5μg mL-1 but at 72 h of treatment. In case of the extract of in vitro origin, the inhibition was found against EAC cell lines even at 48h. During spectrophotometric scanning, the extracts exhibited different maxima (ʎ) - four peaks in in vitro extracts as against single in in vivo preparation suggesting the possible change in the nature of ingredients during micropropagation through tissue culture techniques.

Improved Technique of Non-viral Gene Delivery into Cancer Cells

Liposomal magnetofection is a simple, highly efficient technology for cell transfection, demonstrating better outcome than a number of other common gene delivery methods. However, aggregate complexes distribution over the cell surface is non-uniform due to the gradient of the permanent magnetic field. The aim of this study was to estimate the efficiency of liposomal magnetofection for prostate carcinoma PC3 cell line using newly designed device, “DynaFECTOR", ensuring magnetofection in a dynamic gradient magnetic field. Liposomal magnetofection in a dynamic gradient magnetic field demonstrated the highest transfection efficiency for PC3 cells – it increased for 21% in comparison with liposomal magnetofection and for 42% in comparison with lipofection alone. The optimal incubation time under dynamic magnetic field for PC3 cell line was 5 minutes and the optimal rotation frequency of magnets – 5 rpm. The new approach also revealed lower cytotoxic effect to cells than liposomal magnetofection.

Polymorphic Marker Designed from Bioinformatics Sequences Related to Cell Wall Strength for Discrimination of Mangosteen (Garcinia mangostana L.) Clones Resistant to Gamboge Disorder

Gamboge disorder (GD) or fruit damage by the yellow sap is a major problem in mangosteen. Mangosteen plants varied in the level of GD, from very low or non GD to low, moderate and high GD. However it was difficult to differentiate between GD and non GD plants because evaluation of the disorder is strongly influenced by environment. In this study we investigated the usefulness of primer designed from bioinformatics related to cell wall strength, termed as MCWS, to predict GD. Plant materials used were 28 mangosteen plants selected based on percentage of GD categorized as high, moderate, low and very low or non GD. The result showed that the specific DNA fragments were absent in the high GD accessions. The MCWS marker suggests as a novel polymorphic marker for GD in mangosteen as well as a marker for detect variability in mangosteen as apomictic plant.

Interactions between Cells and Nanoscale Surfaces of Oxidized Silicon Substrates

The importance for manipulating an incorporated scaffold and directing cell behaviors is well appreciated for tissue engineering. Here, we developed newly nano-topographic oxidized silicon nanosponges capable of being various chemical modifications to provide much insight into the fundamental biology of how cells interact with their surrounding environment in vitro. A wet etching technique is exerted to allow us fabricated the silicon nanosponges in a high-throughput manner. Furthermore, various organo-silane chemicals enabled self-assembled on the surfaces by vapor deposition. We have found that Chinese hamster ovary (CHO) cells displayed certain distinguishable morphogenesis, adherent responses, and biochemical properties while cultured on these chemical modified nano-topographic structures in compared with the planar oxidized silicon counterparts, indicating that cell behaviors can be influenced by certain physical characteristic derived from nano-topography in addition to the hydrophobicity of contact surfaces crucial for cell adhesion and spreading. Of particular, there were predominant nano-actin punches and slender protrusions formed while cells were cultured on the nano-topographic structures. This study shed potential applications of these nano-topographic biomaterials for controlling cell development in tissue engineering or basic cell biology research.

Bioefficacy of Some Oil-Mixed Plant Derivatives against African Mud Catfish (Clarias gariepinus) Beetles, Dermestes maculatus and Necrobia rufipes

The efficacy of the separate mixing of four tropical spicy and medicinal plant products: Dennettia tripetala Baker (pepper fruit), Eugenia aromatica Hook (clove), Piper guineense (Schum and Thonn) (black pepper) and Monodora myristica (Dunal) (African nut-meg) with a household vegetable oil was evaluated under tropical storage conditions for the control and reproductive performance of Dermestes maculatus (De Geer) (hide beetle) and Necroba rufipes (De Geer) (copra beetle) on African catfish, Clarias gariepinus (Burchell). Each of the plant materials was pulverized into powder and applied as a mix of 1ml of oil and plant powder at 2.5, 5.0, 7.5 and 10.0g per 100g of dried fish, and allowed to dry for 6h. Each of the four oil-mixed powder treatments evoked significant (P < 05) mortalities of the two insects compared with the control (oil only) at 1, 3 and 7 days post treatment. The oil-powder mixture dosages did not prevent insect egg hatchability but while the emergent larvae on the treated samples died, the emergent larvae in the control survived into adults. The application of oil-mixed powders effectively suppressed the emergence of the larvae of the beetles. Similarly, each of the oil-powder mixtures significantly reduced weight loss in smoked fish that were exposed to D. maculatus and N. rufipes when compared to the control (P < 05). The results of this study suggest that the plant powders rather than the domestic oil demonstrated protective ability against the fish beetles and confirm the efficacy of the plant products as pest control agents.

An Information Theoretic Approach to Rescoring Peptides Produced by De Novo Peptide Sequencing

Tandem mass spectrometry (MS/MS) is the engine driving high-throughput protein identification. Protein mixtures possibly representing thousands of proteins from multiple species are treated with proteolytic enzymes, cutting the proteins into smaller peptides that are then analyzed generating MS/MS spectra. The task of determining the identity of the peptide from its spectrum is currently the weak point in the process. Current approaches to de novo sequencing are able to compute candidate peptides efficiently. The problem lies in the limitations of current scoring functions. In this paper we introduce the concept of proteome signature. By examining proteins and compiling proteome signatures (amino acid usage) it is possible to characterize likely combinations of amino acids and better distinguish between candidate peptides. Our results strongly support the hypothesis that a scoring function that considers amino acid usage patterns is better able to distinguish between candidate peptides. This in turn leads to higher accuracy in peptide prediction.

Detection of Pathogenic Escherichia coli Strains Pollution in Red Deer Meat in Latvia and Determination the Compatibility of VT1, VT2, eae A Genes in their Isolate

Tasks of the work were study the possible E.coli contamination in red deer meat, identify pathogenic strains from isolated E.coli, determine their incidence in red deer meat and determine the presence of VT1, VT2 and eaeA genes for the pathogenic E.coli. 8 (10%) samples were randomly selected from 80 analysed isolates of E.coli and PCR reaction was performed on them. PCR was done both on initial materials – samples of red deer meat - and for already isolated liqueurs. Two of analysed venison samples contain verotoxin-producing strains of E. coli. It means that this meat is not safe to consumer. It was proven by the sequestration reaction of E. coli and by comparison of the obtained results with the database of microorganism genome available on the internet that the isolated culture corresponds to region 16S rDNS of E. coli thus presenting correctness of the microbiological methods.

Sericulture for Employment Generation among the Tribal- A Study of Two Trible Block of Raigarh Dist. [C.G.] India

Among many agro- based cottage industries in India sericulture has been promoted as an agro-based, labor intensive, rural oriented cottage industry, providing gainful employment mainly to the weaker and marginalized section of the society specially tribal. Sericulture occupies the place of pride in the rural economy can be practiced even with very low land holding, low gestation, high returns make sericulture an ideal program, requiring little capital investment. In 2010-2011 the employment in sericulture sector was 72.5 lakh persons. The involvement of landless rural people in tasar sericulture is because they understood its potential for rural and tribal upliftment. This article demonstrates that certain developmental initiatives have been playing an important role in the socio-economic progress of tribal masses in Raigarh district and explains the increased returns from sericulture as a result of development programs. The study concludes with some suggestions to improve the long term feasibility of sericulture.

Allelopathic Effects of Sisymbrium irio L. and Descurainia sophia (L.) Schur on the Germination of Wheat (Triticum aestivum L.)

An experiment was conducted under controlled conditions to study the effect of water extract of leaves, shoots and roots of either Sisymbrium irio L. =SISIR and/or Descurainia sophia (L.) Schur =DESSO on the germination and primary growth of wheat. A split-split plot experiment in CRD with three replications was used. The main plots were the type of weed: i.e. SISIR and DESSO and the sub-plots were type of organ: i.e. leaf, stem and root and, the sub-sub plots were concentration of the water extract of each organ of the weeds: i.e. 0, 2, 4 and 8 % w/v. The plant materials were cut in 2-3 cm pieces and then were ground in a blender. The crushed materials were weighed according to experimental protocol and the final volume was reached to 100 ml in distilled water in dark bottles. All bottles were put on a shaker for 24 hours. The solutions were filtered by muslin cloth. Whatman paper, 9 cm in diameter, were put in petri dishes and twenty seeds of wheat were put on it and 5 ml distilled water or water extract of weeds were added to each petri dish. All petri dishes were put in constant temperature of 15 0C incubator. The results showed that the SISIR water extract had a greater inhibitory effects on germination and primary growth of wheat than those of DESSO water extract. The water extracts of the leaves of both weeds had the greatest inhibitory effects on germination and primary growth of wheat, compared to those of stems and roots. Increasing the concentration of water extract of leaves, stems and roots of both weeds up to 8 % caused the greatest inhibitory effects to wheat and reduced the germination rate and primary growth of it linearly.

Determination of Alkaline Protease Production In Serratia Marcescens Sp7 Using Agro Wastes As Substrate Medium, Optimization Of Production Parameters And Purification Of The Enzyme

The enzyme alkaline protease production was determined under solid state fermentation using the soil bacteria Serratia marcescens sp7. The maximum production was obtained from wheat bran medium than ground nut shell and chemically defined medium. The physiological fermentation factors such as pH of the medium (pH 8), Temperature (40oC) and incubation time (48 hrs) played a vital role in alkaline protease production in all the above. 100Mm NaCl has given better resolution during elution of the enzymes. The enzyme production was found to be associated with growth of the bacterial culture.

Gonadotoxic and Cytotoxic Effect of Induced obesity via Monosodium Glutamate on Mus musculus Testis Cytoarchitecture and Sperm Parameter

Impaired fertility may be the result of indirect consumption of anti-fertility agents through food. Monosodium glutamate (MSG) has been widely used as food additive, flavour enhancer and included in vaccines. This study focuses in determining the gonadotoxic and cytotoxic effect of MSG on selected sperm parameters such as sperm viability, sperm membrane integrity and testes cytoarchitecture of male mice via histological examination to determine its effect on spermatogenesis. Twenty-four Mus musculus were randomly divided into 4 groups and given intraperitoneal injections (IP) daily for 14 days of different MSG concentrations at 250, 500 and 1000mg/kg MSG to body weight to induce obesity. Saline was given to control group. Mice were sacrificed and analysis revealed abnormalities in values for sperm parameters and damages to testes cytoarchitecture of male mice. The results recorded decreased viability (p0.05) with degenerative structures in seminiferous tubule of testes. The results indicated various implications of MSG on male mice reproductive system which has consequences in fertility potential.

Pseudo-Homogeneous Kinetic of Dilute-Acid Hydrolysis of Rice Huskfor Ethanol Production: Effect of Sugar Degradation

Rice husk is a lignocellulosic source that can be converted to ethanol. Three hundreds grams of rice husk was mixed with 1 L of 0.18 N sulfuric acid solutions then was heated in an autoclave. The reaction was expected to be at constant temperature (isothermal), but before that temperature was achieved, reaction has occurred. The first liquid sample was taken at temperature of 140 0C and repeated every 5 minute interval. So the data obtained are in the regions of non-isothermal and isothermal. It was observed that the degradation has significant effects on the ethanol production. The kinetic constants can be expressed by Arrhenius equation with the frequency factors for hydrolysis and sugar degradation of 1.58 x 105 min-1 and 2.29 x 108 L/mole-min, respectively, while the activation energies are 64,350 J/mole and 76,571 J/mole. The highest ethanol concentration from fermentation is 1.13% v/v, attained at 220 0C.

Production of IAA by Bradyrhizobium sp.

The objective of this research was to determine the potency of indigenous acid-aluminium tolerant Bradyrhizobium japonicum as producer of indole acetic acid (IAA) and applied it as nitrogen fixation on local soybeans viz Anjasmoro, Tanggamus (yellow soybean seeds), and Detam (black soybean seed). Three isolates of acid-aluminium tolerant Bradyrhizobium japonicum (BJ) were used in this research, i.e. BJ 11 (wt), BJ 11 (19) - BJ 11(wt) mutant, and USDA 110 as a reference isolate. All of isolates tested to produce the IAA by using Salkowsky method. Effect of IAA production by each of B. japonicum was tested on growth pouch and greenhouse using three varieties of soybean. All isolates could grow well and produce IAA on yeast mannitol broth (YMB) medium in the presence of 0.5 mM L-tryptophan. BJ 11 (19) produced the highest of IAA at 4 days incubation compared to BJ 11 (wt) and USDA 110. All tested isolates of Bradyrhizobium japonicum have showed effect on stimulating the formation of root nodules in soybean varieties grown on Leonard bottle. The concentration of IAA on root nodules of soybean symbiotic with B. japonicum was significantly different with control, except on the treatment using Tanggamus soybean.

Codon-optimized Carbonic Anhydrase from Dunaliella species: Expression and Characterization

Carbonic anhydrases (CAs) has been focused as biological catalysis for CO2 sequestration process because it can catalyze the conversion of CO2 to bicarbonate. Here, codon-optimized sequence of α type-CA cloned from Duneliala species. (DsCAopt) was constructed, expressed, and characterized. The expression level in E. coli BL21(DE3) was better for codon-optimized DsCAopt than intact sequence of DsCAopt. DsCAopt enzyme shows high-stability at pH 7.6/10.0. In final, we demonstrated that in the Ca2+ solution, DsCAopt enzyme can catalyze well the conversion of CO2 to CaCO3, as the calcite form.

The Key Role of the Steroidal Hormones in the Pattern Distribution of the Epiphyseal Structure in Rabbit

Steroidal hormones with the efficient changes on the epiphyseal growth plate may influence tissue structure properties. Presents paper to investigate the effects of gonadectomy in the pattern distribution of the epiphyseal structure. Fifteen adult female New Zealand white rabbits were separated into three groups. One group was intact and others groups were selected for surgical operation. From these two groups, one group carried out steroidal administration. The results obtained showed that there is no statistically difference in the mean diameter of the growth plate cells between all three groups. The maximum value of the cartilage cells were allocated to the gonadectomized group and the minimum number were observed in Hormonal induced group significantly. Growth plate height was significantly greater in gonadectomized group than in two other groups.

The Quality Maintenance and Extending Storage Life of Mango Fruit after Postharvest Treatments

The quality attributes and storage life of 'Jinhwang' mango fruit can be effectively maintained with 1-methylcyclopropene (1-MCP) application and/or chitosan coating. 'Jinhwang' mango fruit was treated with 5 μl l-1 1-MCP for 12 h, dipped with 0.5 % chitosan, 5 μl l-1 1-MCP combine with 0.5 % chitosan and untreated (control) then stored at 10oC. Mango treated with 1-MCP maintained firmness, sucrose and starch content. Chitosan coating delayed firmness loss, sucrose content and the fruit decay when compare with control. Application of 1-MCP combine with chitosan also delayed firmness loss, sucrose content and starch content during storage. Furthermore, chitosan coating and combine treatment prolonged storage life of mango up to 29 days after storage while 1-MCP extended to 28 days after storage. Therefore, using all application of chitosan coating or 1-MCP combine with chitosan or 1-MCP in mango at 10oC is a feasible technology for maintains quality and prolongs storage life in order to expand marketability and export options.

Some Physico-chemical and Nutritional Properties of Russion Olive (Elaeagnus angustifolia L.) Fruit Grown in Turkey

In this research, several physical and chemical properties and mineral contents of Russion olive (Elaeagnus angustifolia L.) fruit grown in Turkey were determined. In the fruit, length, width, thickness, weight were determined as 2.866cm, 1.880cm, 1.870cm, 2.90g; total soluble solids and colour were determined as 13.14% and S33O60Y99, respectively. In the seed, mean length, width, thickness, and weight were found to be 2.420cm, 0.508cm, 0.503cm, and 0.38g, respectively. In the fruit, 28 minerals (K, P, Ca, Na, Mg, S, Pb, Ba, Ga, As, In, Ti, Fe, Mn, Zn, Cu, Co, B, Cd, Se, Li, Sr, Al, Ni, Ag, V, Cr, Bi) were analyzed and 19 minerals were present at detectable levels. Russion olive fruit was richest in potassium (10296.906ppm), Mg (762.314ppm), P (609.694ppm), Ca (547.647ppm), and Na (222.749ppm).

Determination of Yield and Some Quality Characteristics of Winter Canola (Brassica napus ssp. oleifera L.) Cultivars

Canola is a specific edible type of rapeseed, developed in the 1970s, which contains about 40 percent oil. This research was carried out to determine the yield and some quality characteristics of some winter canola cultivars during the 2010-2011 vegetation period in Central Anatolia of Turkey. In this research; Oase, Dante, Californium, Excalibur, Elvis, ES Hydromel, Licord, Orkan, Vectra, Nelson, Champlain and NK Petrol winter canola varieties were used as material. The field experiment was set up in a “Randomized Complete Block Design” with three replications on 21 September 2010. In this research; seed yield, oil content, protein content, oil yield and protein yield were examined. As a result of this research; seed yield, oil content, oil yield and protein yield (except protein content) were significant differences between the cultivars. The highest seed yield (6348 kg ha-1) was obtained from the NK Petrol, while the lowest seed yield (3949 kg ha-1) was determined from the Champlain cultivar was obtained. The highest oil content (46.73%) was observed from Oase and the lowest value was obtained from Vectra (41.87%) cultivar. The highest oil yield (2950 kg ha-1) was determined from NK Petrol while the least value (1681 kg ha-1) was determined from Champlain cultivar. The highest protein yield (1539.3 kg ha-1) was obtained from NK Petrol and the lowest protein yield (976.5 kg ha-1) was obtained from Champlain cultivar. The main purpose of the cultivation of oil crops, to increase the yield of oil per unit area. According the result of this research, NK Petrol cultivar which ranks first with regard to both seed yield and oil yield between cultivars as the most suitable winter canola cultivar of local conditions.

Full-genomic Network Inference for Non-model organisms: A Case Study for the Fungal Pathogen Candida albicans

Reverse engineering of full-genomic interaction networks based on compendia of expression data has been successfully applied for a number of model organisms. This study adapts these approaches for an important non-model organism: The major human fungal pathogen Candida albicans. During the infection process, the pathogen can adapt to a wide range of environmental niches and reversibly changes its growth form. Given the importance of these processes, it is important to know how they are regulated. This study presents a reverse engineering strategy able to infer fullgenomic interaction networks for C. albicans based on a linear regression, utilizing the sparseness criterion (LASSO). To overcome the limited amount of expression data and small number of known interactions, we utilize different prior-knowledge sources guiding the network inference to a knowledge driven solution. Since, no database of known interactions for C. albicans exists, we use a textmining system which utilizes full-text research papers to identify known regulatory interactions. By comparing with these known regulatory interactions, we find an optimal value for global modelling parameters weighting the influence of the sparseness criterion and the prior-knowledge. Furthermore, we show that soft integration of prior-knowledge additionally improves the performance. Finally, we compare the performance of our approach to state of the art network inference approaches.

Evaluating the Response of Rainfed-Chickpea to Population Density in Iran, Using Simulation

The response of growth and yield of rainfed-chickpea to population density should be evaluated based on long-term experiments to include the climate variability. This is achievable just by simulation. In this simulation study, this evaluation was done by running the CYRUS model for long-term daily weather data of five locations in Iran. The tested population densities were 7 to 59 (with interval of 2) stands per square meter. Various functions, including quadratic, segmented, beta, broken linear, and dent-like functions, were tested. Considering root mean square of deviations and linear regression statistics [intercept (a), slope (b), and correlation coefficient (r)] for predicted versus observed variables, the quadratic and broken linear functions appeared to be appropriate for describing the changes in biomass and grain yield, and in harvest index, respectively. Results indicated that in all locations, grain yield tends to show increasing trend with crowding the population, but subsequently decreases. This was also true for biomass in five locations. The harvest index appeared to have plateau state across low population densities, but decreasing trend with more increasing density. The turning point (optimum population density) for grain yield was 30.68 stands per square meter in Isfahan, 30.54 in Shiraz, 31.47 in Kermanshah, 34.85 in Tabriz, and 32.00 in Mashhad. The optimum population density for biomass ranged from 24.6 (in Tabriz) to 35.3 stands per square meter (Mashhad). For harvest index it varied between 35.87 and 40.12 stands per square meter.