Identifying the presence of bacteria on digital images by using asymmetric distribution with k-means clustering algorithm.

This paper is mainly aimed at the decomposition of image quality assessment study by using Three Parameter Logistic Mixture Model and k-means clustering (TPLMM-k). This method is mainly used for the analysis of various images which were related to several real time applications and for medical disease detection and diagnosis with the help of the digital images which were generated by digital microscopic camera. Several algorithms and distribution models had been developed and proposed for the segmentation of the images. Among several methods developed and proposed, the Gaussian Mixture Model (GMM) was one of the highly used models. One can say that almost the GMM was playing the key role in most of the image segmentation research works so far noticed in the literature. The main drawback with the distribution model was that this GMM model will be best fitted with a kind of data in the dataset. To overcome this problem, the TPLMM-k algorithm is proposed. The image decomposition process used in the proposed algorithm had been analyzed and its performance was analyzed with the help of various performance metrics like the Variance of Information (VOI), Global Consistency Error (GCE) and Probabilistic Rand Index (PRI). According to the results, it is shown that the proposed algorithm achieves the better performance when compared with the previous results of the previous techniques. In addition, the decomposition of the images had been improved in the proposed algorithm.

MONO, DI and TRI SSRs data extraction & storage from 1403 virus genomes with next generation retrieval mechanism.

Now a day׳s SSRs occupy the dominant role in different areas of bio-informatics like new virus identification, DNA finger printing, paternity & maternity identification, disease identification, future disease expectations and possibilities etc., Due to their wide applications in various fields and their significance, SSRs have been the area of interest for many researchers. In the SSRs extraction, retrieval algorithms are used; if retrieval algorithms quality is improved then automatically SSRs extraction system will achieve the most relevant results. For this retrieval purpose in this paper a new retrieval mechanism is proposed which will extracted the MONO, DI and TRI patterns. To extract the MONO, DI and TRI patterns using proposed retrieval mechanism in this paper, DNA sequence of 1403 virus genome data sets are considered and different MONO, DI and TRI patterns are searched in the data genome sequence file. The proposed Next Generation Sequencing (NGS) retrieval mechanism extracted the MONO, DI and TRI patterns without missing anything. It is observed that the retrieval mechanism reduces the unnecessary comparisons. Finally the extracted SSRs provide the useful, single view and useful resource to researchers.

Chemical and pharmacological standardization of Ashwagandhadi lehyam: an ayurvedic formulation.

Standardization is an important measurement for ensuring the quality control of herbal drugs. It has become essential to develop reliable, specific and sensitive quality control methods. One of the prime ingredients of Ashwagandhadi lehyam is Withania somnifera L. Dunal (Solanaceae) (ashwagandha). In the present study, Ashwagandhadi lehyam was formulated and the quality assessment of the formulation was based on phytochemical screening and physico-chemical evaluation. Ashwagandhadi lehyam was formulated according to a working formula and subjected to phytochemical screening by FTIR analysis and HPTLC fingerprinting, heavy metal determination by AAS, determination of alcohol content, tested for Escherichia coli, Staphylococcus aureus, aerobic bacteria, yeasts and mould, oral toxicity studies and anti-epileptic activity by MES method. The physico-chemical studies showed total ash content as 6.45%, extractive values and some trace elements such as lead, mercury, cadmium and arsenic with 3.2, 0.05, 0.18 and 0.48 ppm, respectively. FTIR and HPTLC studies revealed the presence of functional groups of withanolides in Ashwagandhadi lehyam, resulting in its chemical standardization. The formulation exhibited less epileptic seizures in various phases when compared with that of standard phenytoin and found to possess better anti-inflammatory activity, thus making it biologically standardized. The physico-chemical and pharmacological analysis to standardize Ashwagandhadi lehyam confirmed its use as a safe anti-inflammatory agent and for various seizure disorders.

Clinical evaluation of intrabony defects in localized aggressive periodontitis patients with and without bioglass- an in-vivo study.

BACKGROUND: Aggressive periodontitis is a specific type of periodontitis with clearly identifiable clinical characteristics such as rapid attachment loss, bone destruction and familial aggregation. Regeneration of mineralized tissues affected by aggressive periodontitis comprises a major scientific and clinical challenge. In recent years some evidence has been provided that bioactive glass is also capable of supporting the regenerative healing of periodontal lesions. OBJECTIVE: The aim of this clinical and radiological prospective study was to evaluate the efficacy of bioactive glass in the treatment of intra-bony defects in patients with localized aggressive periodontitis. METHODS: Twelve localized aggressive periodontitis patients with bilaterally located three-walled intra-bony defect depth = 2 mm, preoperative probing depths = 5 mm were randomly treated either with the bioactive glass or without the bioactive glass. The clinical parameters plaque index, gingival index, probing depth, gingival recession, clinical attachment level, and mobility were recorded prior to surgery as well as 12 months after surgery. Intraoral radiographs were digitized to evaluate the bone defect depth at baseline and 12 months after the surgery. RESULTS: After 12 months, a reduction in probing depth of 3.92 + 0.313 mm (P < 0.001) and a gain in clinical attachment level of 4.42+0358mm (P < 0.001) were registered in the test group. In the control group, a reduction in probing depth of 2.5 +0.230mm (P <0.001) and a gain in clinical attachment level of 2.58 + 0.149 mm (P <0.001) was recorded. Radiographically, the defects were found to be filled by 2.587 + 0.218 mm (P < 0.001) in the test group and by 0.1792 + 0.031mm (P < 0.001) in the control group. Changes in gingival recession showed no significant differences. . CONCLUSION: Highly significant improvements in the parameters Probing depth, Clinical attachment level, and Bone defect depth were recorded after 12 months, with regenerative material.

Evaluation of PCR-based methods for isolating flanking regions of genes.

Several polymerase chain reaction (PCR)-based methods are available for isolation of unknown genomic fragments. In the present study, a comparative evaluation of a few methods of ligation-mediated PCR methods and a ligation-independent one were made by isolating promoter fragment for N-methyltransferase gene involved in the caffeine biosynthetic pathway of Coffea canephora. The benefits of tertiary PCR and the effects of a 4-base cutting restriction endonuclease on the size of the PCR products obtained were demonstrated in one of the ligation-mediated PCR methods. The methods adopted in this study differed in the sizes of the 5'-flanking regions obtained. The efficiencies of various methods used reflect the inherent limitations of the PCR-based methods for isolation of unknown flanking regions.

Stable transformation and direct regeneration in Coffea canephora P ex. Fr. by Agrobacterium rhizogenes mediated transformation without hairy-root phenotype.

A system for genetic transformation of Coffea canephora by co-cultivation with Agrobacterium rhizogenes harbouring a binary vector has been developed. The objective of the present study was the genetic transformation and direct regeneration of transformants through secondary embryos bypassing an intervening hairy root stage. Transformants were obtained with a transformation efficiency up to 3% depending on the medium adjuvant used. A. rhizogenes strain A4 harbouring plasmid pCAMBIA 1301 with an intron uidA reporter and hygromycin phosphotransferase (hptII) marker gene was used for sonication-assisted transformation of Coffea canephora. The use of hygromycin in the secondary embryo induction medium allowed the selection of transgenic secondary embryos having Ri T-DNA along with the T-DNA from the pCAMBIA 1301 binary vector. In addition transgenic secondary embryos devoid of Ri-T-DNA but with stable integration of the T-DNA from the binary vector were obtained. The putative transformants were positive for the expression of the uidA gene. PCR and Southern blot analysis confirmed the independent, transgenic nature of the analysed plants and indicated single and multiple locus integrations. The study clearly demonstrates that A. rhizogenes can be used for delivering transgenes into tree species like Coffea using binary vectors with Agrobacterium tumefaciens T-DNA borders.

Isolation of promoter for N-methyltransferase gene associated with caffeine biosynthesis in Coffea canephora.

N-Methyltransferases (NMTs) catalyze the three SAM dependent sequential methylation of xanthosine, producing caffeine in Coffea species. In the present work, a PCR based genome walking method was adopted to isolate and clone the promoter for the NMT gene. Inspection of the promoter sequence revealed the presence of several motifs important for the regulation of the gene expression. The whole fragment was fused to the beta-glucuronidase (gus) reporter gene and used in Agrobacterium tumefaciens mediated transformation of Nicotiana tabacum. GUS assays proved that the isolated promoter was able to direct the expression of the reporter gene in transgenic tobacco. Based on the promoter sequence, primer was designed and the genomic fragment comprising the promoter and its corresponding gene was amplified and cloned. Sequencing of one of the genomic clones revealed the presence of four exons and three introns in NMT gene. The differences in the restriction pattern among the genomic clones were studied using PCR-RFLP. This is the first report of cloning of the promoter for a gene involved in caffeine biosynthetic pathway and it opens up the possibility of studying the molecular mechanisms that regulate the production of caffeine.

A new tetracyclic triterpene from the latex of Euphorbia nerifolia.

A new tetracyclic triterpene 9,19-cyclolanost-22(22'), 24-diene-3beta-ol, named as nerifoliene (2) along with euphol (1) were isolated from the fresh latex of Euphorbia nerifolia. Their structures were elucidated on the basis of spectral (IR, 1H and 13C NMR, FAB and EI Mass) data.

A study of morphology, cytology and sterility in interspecific hybrids and amphidiploids of Nicotiana knightiana X N. umbratica.

Interspecific hybrids and amphidiploids of Nicotiana knightiana Goodspeed (n= 12)x N. umbratica Burbidge (n = 23) resembled either parent in some characters and were intermediate in other characters. The F1 hybrids (2n = 35) showed mostly univalents during meiosis, while the amphidiploids (2n = 70) formed bivalents almost regularly. The former were completely sterile and the latter fully male fertile but predominantly female sterile. This female sterility was due to disintegration of the embryo sacs leading to collapsed ovules. The few fertile ovules, however, showed normal development of embryo sac and embryo. The occurrence of fertile and sterile ovules was believed to be due to segregation of the genes governing sterility.

R-stippled maize as a transposable element system.

The I-R element at the R locus destabilizes kernel pigmentation giving the variegated pattern known as stippled ( R-st). In trans linkage phase with R-st the element was shown to act as a modifier of stippled, intensifying seed spotting in parallel with effects of the dominant linked modifier M-st. Presence of I-R in the genome was, therefore, shown to be detectable as a modifier of R-st. When this test was used, new modifiers resembling M-st were often detected following mutations of R-st to the stable allele R-sc. Such mutations evidently occurred by transposition of I-R away from the R locus to a site where it was identifiable as a modifier. M-st may be such a transposed I-R. Analysis of mutations to R-sc during the second (sperm-forming) mitosis in pollen grains showed that some of the transposed I-R elements were linked with R, whereas others assorted independently. Their strengths varied from barely discernible to a level equal to M-st. Overreplication frequently accompanied transposition at the sperm-forming mitosis, leading to transposed I-R elements in both the mutant and nonmutant sperm.

Cytogenetic study of an interspecific cross of Nicotiana debneyi X N. umbratica.

Interspecific F1 hybrids of Nicotiana debneyi Domin (2n=48) and N. umbratica Burbidge (2n=46), both belonging to the section Suaveolentes, showed a high degree of meiotic chromosome pairing. Two of the five F2 plants obtained exhibited chromosome mosaicism. The first colchiploid generation (C1) had the expected chromosome number of 2n=94 while C2 showed 2n=88, a loss of three pairs of chromosomes. This same chromosome number continued in further colchiploid generations, followed up to C5, except for a few plants in C3 which showed chromosome mosaicism. The F1 phenotype was stable through C1 to C5 and fertility was normal in colchiploids through all generations in spite of the loss of three pairs of chromosomes and chromosome mosaicism. This stability and fertility apparently reflect the tolerance of the genomes to the genetic adjustment of chromosome complements which is believed to be associated with the originally polyploid nature of the parental species and the chromosome doubling brought about in the amphidiploids.