Environmental microbes like Bordetella petrii has been established as a causative agent for various infectious diseases in human. Again, development of drug resistance in B. petrii challenged to combat against the infection. Identification of potential drug target and proposing a novel lead compound against the pathogen has a great aid and value. In this study, bioinformatics tools and technology have been applied to suggest a potential drug target by screening the proteome information of B. petrii DSM 12804 (accession No. PRJNA28135) from genome database of National Centre for Biotechnology information. In this regards, the inhibitory effect of nine natural compounds like ajoene (Allium sativum), allicin (A. sativum), cinnamaldehyde (Cinnamomum cassia), curcumin (Curcuma longa), gallotannin (active component of green tea and red wine), isoorientin (Anthopterus wardii), isovitexin (A. wardii), neral (Melissa officinalis), and vitexin (A. wardii) have been acknowledged with anti-bacterial properties and hence tested against identified drug target of B. petrii by implicating computational approach. The in silico studies revealed the hypothesis that lpxD could be a potential drug target and with recommendation of a strong inhibitory effect of selected natural compounds against infection caused due to B. petrii, would be further validated through in vitro experiments.
Garcia, Liliana Torcoroma;Cristancho, Laura Maritza;Vera, Erika Patricia;Begambre, Oscar
Journal of Microbiology and Biotechnology
/
v.25
no.10
/
pp.1714-1727
/
2015
This work describes a new strategy for optimal design of Multiplex-PCR primer sequences. The process is based on the Particle Swarm Optimization-Simplex algorithm (Mult-PSOS). Diverging from previous solutions centered on heuristic tools, the Mult-PSOS is selfconfigured because it does not require the definition of the algorithm's initial search parameters. The successful performance of this method was validated in vitro using Multiplex-PCR assays. For this validation, seven gene sequences of the most prevalent bacteria implicated in urinary tract infections were taken as DNA targets. The in vitro tests confirmed the good performance of the Mult-PSOS, with respect to infectious disease diagnosis, in the rapid and efficient selection of the optimal oligonucleotide sequences for Multiplex-PCRs. The predicted sequences allowed the adequate amplification of all amplicons in a single step (with the correct amount of DNA template and primers), reducing significantly the need for trial and error experiments. In addition, owing to its independence from the initial selection of the heuristic constants, the Mult-PSOS can be employed by non-expert users in computational techniques or in primer design problems.
The effect of diets varying in level and source of nitrogen (N) and fermentable organic matter on dynamic characteristics of microbial populations in rumen liquor and their impact on substrate fermentation in vitro was studied. The diets tested were straw alone, straw+concentrate mixture and straw+urea molasses mineral block (UMMB) lick. The same diets were taken as substrates and tested on each inoculum collected from the diets. Diet had no effect on the amino acid (AA) composition of either bacteria or protozoa. Differences among the diets in intake, source of N and OM affected bacterial and protozoal characteristics in the rumen. Upper asymptote of gas production (Y$\alpha$) had a higher correlation with bacterial pool size and production rate than with protozoal pool size and production rate. Among the parameters of the gas production model, Y$\alpha$ and lag time in total gas has showed significant (p<0.01) correlation with bacterial characteristics. Though the rate constant of gas production significantly differed (p<0.01) between diet and type of straw, it was least influenced by the microbial characteristics. The regression coefficient of diet and type of straw for Y$\alpha$ indicated that the effect of diet on Y$\alpha$ was threefold higher than that of the straw. As microbial characteristics showed higher correlation with Y$\alpha$, and diet had more influence on the microbial characteristics, gas production on a straw diet could be used effectively to understand the microbial characteristics.
As described here, most recently developed methods for improving reproduction performance of domesticated animals such as cattle, swine and chicken have been considered to be also usable for restoring some sorts of endangered and/or extinct wild animals in the very near future. Especially, the techniques for in vitro storage of gametes obtained from dead animals shortly after the death, probably 24 h following the sacrifice are also available for obtaining some of experimental specimens. In case of the endangered animals, nobody will be allowed to use any tissues from the living animals, therefore, e.g., the use of skin tissues from these bodies is another possibility of restoring the living animals. Regarding the use of skin tissues, the most highly usable tools must be the cloning techniques for reviving rare cells from the living body. Most possible techniques for cloning cells is nuclear transfer from rare species to highly relative species, and this is the case of germ cells, e.g., primordial germ cells (PGCs) of avian species. One of the possibilities is the nuclear transfer of Crested Ibis (Nipponia nippon) to the PGCs of chicken, resulting in the PGCs with transferred nucleus from the ibis. In mammalian species, the same procedure as in the case of birds would be successful, e.g., the removed nucleus from Giant Pandas will be transferred to the cell, such as somatic cells or germ cells from black bears or lesser pandas, leading to the production of transnucleared cells in the body of female black bears. These two cases are most promising techniques for reviving endangered animals in the world, particularly in Asian countries, mainly in China. As a conclusion, possible production of cloned animals carrying transnucleared cells from endangered animals, such as Giant Pandas and Crested Ibis, may be reproduced gradually in the near future. Scientists are, therefore, required to convert the paradigm from domestic animals to wild animals, including endangered and/or extinct animals on the earth.
Nutritional evaluation of some agro-industrial byproducts available in Samoa [dry brewers' grains (DBG), cocoa shell (CS), cocoa dust (CD) and desiccated coconut waste meal (DCWM)] available in Samoa was carried out using both the in vivo and in vitro techniques. In the in vivo study 24 Anglo-nubian goats were offered by-products with other feed ingredients to compound four different diets. The goats were randomly allocated to 4 diets on the basis of liveweight (18.7-0.3kg). The ADF content of the byproducts followed a similar trend to NDF. The byproducts have a high content of organic matter (91.0-95.4%). Gross energy (GE) content was higher in DCWM (25.1 MJ/kg DM), closely followed by CD (23.2 MJ/kg DM). Concentrate intake was significantly different (p<0.05) among the goats. Average daily live weight gains were 105, 92, 88 and 97 g/goat/day for DBG, CS, CD and DCWM, respectively. Daily live weight gains were higher (p<0.05) in the goats that received DBG, while the least gain was obtained in the goats that received CS byproduct diet. DM digestibility was significantly higher (p<0.05) in the goats on DBG diet than in the other goats. The least DM digestibility was obtained in the goats that received CD diet (p>0.05). CP digestibility followed a similar pattern to DM digestibility. The digestibility of NDF and ADF was influenced by the nature of the diets. The digestibility of OM and GE were best (p<0.05) in the goats that received DBG, DCWM and CS byproduct diets than in CD. Significant differences (p<0.05) among the byproducts were recorded for net gas production. Potential gas production (a+b) ranged from 7.064 to 42.17 ml. Organic matter digested (OMD) from gas production value at 24 h was higher in DBG (47.6 g/kg DM) and this was followed by DCWM (42.5 g/kg DM). The least OMD was obtained in CD (17.9 g/kg DM). A significant difference (p<0.05) in DM disappearance after 4, 8, 16, 24, 48 and 72 h was recorded. The potential and effective degradability varied significantly (p<0.05) from 85.95-99.6 g/kg DM and from 39.9-65.8%, respectively. The digestibility of the byproducts in both the in vivo and in in vitro techniques demonstrated that they are potential source of feed ingredients for ruminant livestock in Samoa and possibly in the other small Pacific Island countries. On the basis of their potential degradability the byproducts could be ranked in the following order:DCWM>DBG>CD>CS. In conclusion, the results obtained suggest that all the byproducts can contribute to ruminant livestock diets without adverse effects on feed intake, growth rate and apparent nutrient digestibility coefficients.
Forestry sector has witnessed some unprecedented events in the recent past both in terms of galloping biotechnological developments and heated environmental debates over risks associated with release of transgenic trees. Improvements in the in vitro propagation techniques has made it possible to develop tissue culture based plant regeneration protocols just for about any tree species. And with the inclusion of every new species within the realms of tissue culture technology, it becomes a candidate for genetic improvement through recombinant DNA technology, the so called genetic engineering. Poplars and their hybrids serve as the model tree species on which most of the genetic transformation work as been carried out. A lot of work has also gone in genetic transformation of fruit trees and trees of horticultural interests. Trees have been successfully transformed for traits ranging from reduction of length of juvenile phase to alteration of tree architecture to altering wood quality by lignin and cellulose modification. More-over trees have been genetically engineered successfully to combat various types of insect pests and pathogens causing diseases. But all these developments have ignited controversies over the possible benefits and risks associated with transgenic plantations by various environmental agencies and activists. Solutions to most of these concerns can be found out with more intensive prioritized research.
The aim of this in vitro study was to investigate the coronal leakage of obligate anaerobes into root canals obturated with two different techniques. 48 extracted human teeth with straight, single root canals were prepared with crown-down technique with Profile under copious irrigation until the master apical file was size 40. The teeth were divided randomly into experimental groups (40 teeth) and control groups (8 teeth). In the experimental groups, 20 teeth were obturated with lateral condensation and other 20 teeth were obturated with continuous wave technique with System B. Coronal leakage of two root canal filing technique was evaluated using anaerobic bacterial leakage model with Fusobacterium nucleatum(ATCC 25586) for 60 days. The results were as follows 1. The incidence of bacterial leakage of experimental groups was 65% in group 1 (lateral condensation) and 60% in group 2 (continuous wave technique with System B). This difference was not statistically significant (P>0.05). 2. There was no statistically significant difference(P>0.05) in leakage score between group 1 (lateral condensation) and group2 (continuous wave technique with System B).
Nguyen Thi Hang;Nguyen Thi Bich Thu;Le Ba Vinh;Nguyen Viet Phong;Tran Van On;Ki Yong Lee
Journal of Microbiology and Biotechnology
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v.33
no.7
/
pp.941-948
/
2023
Metabolites from medicinal plants continue to hold significant value in the exploration and advancement of novel pharmaceuticals. In the search for plants containing compounds with anti-inflammatory effects, we observed that the ethanol (EtOH) extract obtained from the aerial components of Gouania leptostachya DC. var. tonkinensis Pit. exhibited substantial suppression of nitric oxide (NO) in vitro. In a phytochemical study on an EtOH extract of G. leptostachya, 11 compounds were purified, including one unreported compound namely gouanioside A (1). Their chemical structures were unambiguously determined through the use of various spectroscopic techniques, such as 1 and 2D NMR, IR, and HR-ESI-MS, and by producing derivatives via chemical reactions. The EtOH extract, fractions, and a new compound exerted inflammatory effects by altering NO synthesis in murine RAW264.7 macrophage cells stimulated with lipopolysaccharide. The underlying inflammatory mechanism of the new compound 1 was also explored through various in vitro experiments. The results of this study indicate the potential usefulness of new compound 1 from G. leptostachya as a treatment for inflammatory diseases.
Seven essential oils were tested for in vitro cytotoxicity against the cancerous cell lines A-549, HEp-2 and DLA and normal BRL-3A, NRK-49F and Vero cell lines using standard MTT, SRB and dye exclusion techniques. The A-549 cell line was found to be the most susceptible to all the essential oils. The essential oils of A. nilagirica, A. calamus and O. sanctum were found to be the more active against these cells with mean $CTC_{50}$ values of 17.75, 19.00 and $24.37\;{\mu}g/ml$, respectively. The essential oil of Acorus calamus was found to be the most potent with low $CTC_{50}$ values against the cancerous and comparatively higher $CTC_{50}$ values against the normal cell lines. Artemisia pellens and Pelargonium graveolens oils also showed potent activity. These oils merit further investigation to identify the active principles and nature of the anti tumor activity in animal models.
We determined the effects of follicular fluid fractions in the maturation medium on bovine oocyte maturation, fertilization and subsequent development, as well as on number of cells in blastocysts following culture. Follicular fluid and oocytes from bovine follicles less than 5 mm in diameter were collected from the ovaries of slaughtered cows. Follicular fluid was separated into different molecular weight fractions by untrafiltration through a membrane using a centrifuge at 500$\times$g, for 2h. For the maturation medium, follicular fluid fractions (30%, v/v), whole fluid (30%) or PVP(3mg/ml) were added to TCM 199(0.1$\mu\textrm{g}$/ml estradiol-17$\beta$, 100IU hCG). After maturation for 24h, oocytes were fertilized in vitro with bull frozen-thawed spermatozoa and cultured on a monolayer of granulosa cells for 9 days after fertilization. There were no differences in maturation rates or fertilization rates among any maturation conditions. The rates of development to >2-cell stage of the oocytes were significantly decreased when fraction of follicular fluid below 10,000 MW were added into maturation medium, compared with control and fraction above 10,000 MW(26.0% vs 40.8% to 64.0%, respectveily. p<0.01). Likewise, the rates of development to blastocysts of fertilized oocytes were significantly decreased in maturation medium containing fraction of follicular fluid (<10,000 MW). The average cell number of blastocysts derived from oocytes that matured in the fraction(>10,000 MW) of follicular fluid was 154.7$\pm$13.7. These embryos contained more cells than those matured in whole follicular fluid, or the fraction(<10, 000 MW) of follicular fluid or control(107.0$\pm$8.4, 91.8$\pm$11.8 and 95.8$\pm$6.2, respectively). In conclusion, we found that fractions of follicular fluid contained factors stimulating or inhibiting oocyte cytoplasmic matruation. These suggest that a factor(s) inducing cytoplasmic maturation of oocytes may exist in >10,000 MW fraction of follicular fluid.
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