Mining the Proteome of Fusobacterium nucleatum subsp. nucleatum ATCC 25586 for Potential Therapeutics Discovery: An In Silico Approach

The plethora of genome sequence information of bacteria in recent times has ushered in many novel strategies for antibacterial drug discovery and facilitated medical science to take up the challenge of the increasing resistance of pathogenic bacteria to current antibiotics. In this study, we adopted subtractive genomics approach to analyze the whole genome sequence of the Fusobacterium nucleatum, a human oral pathogen having association with colorectal cancer. Our study divulged 1,499 proteins of F. nucleatum, which have no homolog's in human genome. These proteins were subjected to screening further by using the Database of Essential Genes (DEG) that resulted in the identification of 32 vitally important proteins for the bacterium. Subsequent analysis of the identified pivotal proteins, using the Kyoto Encyclopedia of Genes and Genomes (KEGG) Automated Annotation Server (KAAS) resulted in sorting 3 key enzymes of F. nucleatum that may be good candidates as potential drug targets, since they are unique for the bacterium and absent in humans. In addition, we have demonstrated the three dimensional structure of these three proteins. Finally, determination of ligand binding sites of the 2 key proteins as well as screening for functional inhibitors that best fitted with the ligands sites were conducted to discover effective novel therapeutic compounds against F. nucleatum.

Comparative investigation of the free radical scavenging potential and anticancer property of Diospyros blancoi (Ebenaceae)

Objective: To investigate the comparative effects of Diospyros blancoi (Ebenaceae) leaves (DBL), root bark (DBRB) and stem bark (DBSB) on free radicals and cancer. Methods: The polyphenol contents, antioxidant and free radical scavenging properties were determined using standard spectrophotometric methods. Cytotoxicity and anticancer activities were performed on brine shrimp nauplii and Ehrlich ascite carcinoma cells, respectively. Results: Among the extracts, DBSB showed the highest total antioxidant capacity and reducing capacity on ferrous ion. Based on 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radical scavenging activities, DBSB showed (95.760 ± 0.343)% and (67.460 ± 2.641)% scavenging with IC

In Silico Structural and Functional Annotation of Hypothetical Proteins of Vibrio cholerae O139

In developing countries threat of cholera is a significant health concern whenever water purification and sewage disposal systems are inadequate. Vibrio cholerae is one of the responsible bacteria involved in cholera disease. The complete genome sequence of V. cholerae deciphers the presence of various genes and hypothetical proteins whose function are not yet understood. Hence analyzing and annotating the structure and function of hypothetical proteins is important for understanding the V. cholerae. V. cholerae O139 is the most common and pathogenic bacterial strain among various V. cholerae strains. In this study sequence of six hypothetical proteins of V. cholerae O139 has been annotated from NCBI. Various computational tools and databases have been used to determine domain family, protein-protein interaction, solubility of protein, ligand binding sites etc. The three dimensional structure of two proteins were modeled and their ligand binding sites were identified. We have found domains and families of only one protein. The analysis revealed that these proteins might have antibiotic resistance activity, DNA breaking-rejoining activity, integrase enzyme activity, restriction endonuclease, etc. Structural prediction of these proteins and detection of binding sites from this study would indicate a potential target aiding docking studies for therapeutic designing against cholera.

Preparation of Metformin Hydrochloride Extended Release Matrix Tablets by Direct Compression Method and Its In vitro Evaluation.

Aims: Metformin Hydrochloride, a biguanide, is an orally active antihyperglycemic agent, used in the treatment of non-insulin dependent diabetes mellitus (NIDDM). It has relatively short plasma half life, low absolute bioavailability. Extended release formulation of Metformin Hydrochloride by direct compression method has significant challenges due to its poor inherent compressibility and high dose. The aim of this study was to develop extended release tablets of Metformin Hydrochloride by direct compression method and In vitro evaluation. Study Design: Nine different formulations were made by varying drug-polymer ratio and were subjected to different physical property tests of the powder blend as well as prepared tablets, followed by dissolution test. Place and Duration of Study: Department of Pharmacy, State University of Bangladesh, Dhaka, Bangladesh, between January 2013 and July 2013. Methodology: Nine formulations of Metformin Hydrochloride matrix tablets - F-1, F-2, F- 3, F-4, F-5, F-6, F-7, F-8 and F-9 - were prepared by direct compression method using release retarding materials, Methocel K100 MCR Premium (derivative of hydroxypropyl methylcellulose - HPMC) and Xanthan gum. The drug and polymer ratio were 1:0.41, 1:0.45, 1:0.49, 1:0.59, 1:0.63, 1:0.67, 1:0.77, 1:0.81 & 1:0.85 respectively. The micromeritic behavior of the powder blends were evaluated for bulk density, angle of repose, compressibility index along with post compressional attributes of the tablets such as thickness, hardness, friability, weight variation and content of Metformin Hydrochloride in the tablets. The in-vitro drug release study was carried out in 1000 mL phosphate buffer medium (pH 6.8) at 37±0.5°C at 100 rpm for 10 hours using USP Apparatus Type-II (paddle) method. Results: FT-IR study showed drug-excipient compatibility and DSC analysis showed no solid state interaction between components. The physical properties of the powder blend and the tablets were within the acceptable limits. Maximum and minimum drug release were found in formulation F-1 and F-9 respectively which indicate that release rate is inversely proportional to the concentration of Methocel K100 MCR Premium and Xanthan gum in combination. Dissolution study also showed that, formulations F-7, F-8 & F-9 do not comply with drug release specification of USP and among the rest six formulations F- 3, F-4 & F-5 comply better with drug release specification of USP. After fitting the data to Korsmeyer-Peppas equation we found that diffusion along with erosion could be the mechanism of drug release.Considering the micromeritic behaviour of the powder blend, physical attributes of the compressed tablets, and dissolution, formulation F-4 seemed most suitable. Conclusion: Extended release Metformin Hydrochloride tablets can be produced to overcome frequent dosing related problems. However, Further study on formulation optimization and scale up, stability and bioequivalence is needed to confirm the appropriateness of these formulated extended release tablets.

Antioxidant, antibacterial and cytotoxic effects of the phytochemicals of whole Leucas aspera extract

<p><b>OBJECTIVE</b>To investigate the antioxidant, antibacterial and cytotoxic activity of whole Leucas aspera (Labiatae) (L. aspera) alcoholic extract.</p><p><b>METHODS</b>Whole L. aspera powder was extracted by absolute ethanol (99.50%). The ethanolic extract was subjected to antioxidant, antibacterial and brine shrimp lethality assay.</p><p><b>RESULTS</b>The extract showed potent radical scavenging effect (antioxidant) with IC50 value of (99.58±1.22) µg/mL which was significant (P<0.01) in comparison to ascorbic acid with IC50 value of (1.25±0.95) µg/mL. In case of antibacterial screening, the extract showed notable antibacterial effect against the tested microbial strains. Significant (P<0.05) zone of inhibitions against Gram positive Bacillus subtilis [(12.00±1.32) mm] and Bacillus megaterium [(13.00±1.50) mm], Staphylococcus aureus [(8.00±0.50) mm] and Gram negative Salmonella typhi [(6.00±0.50) mm], Salmonella paratyphi [(8.00±1.00) mm], Shigella dysenteriae [(9.00±1.32) mm] and Vibrio cholerae [(9.00±0.66) mm] was observed. In brine shrimp lethality bioassay, the extract showed the LC50 value as (181.68±2.15) µg/mL which was statistically significant (P<0.01) compared to positive control vincristine sulfate [LC50=(0.76±0.04) µg/mL].</p><p><b>CONCLUSIONS</b>The results demonstrate that the ethanolic extract of L. aspera could be used as antibacterial, pesticidal and various pharmacologic actives.</p>

Immunoglobulin levels in panic disorder patients

The study was conducted to evaluate the serum immunoglobulin levels in patients suffering from panic disorder and to assess the relationship between the changes of immunoglobulin levels and the socioeconomic parameters, as well as nutritional status. 54 panic patients were randomly selected from the Department of Psychiatry, Bangabandhu Sheikh Mujib Medical University [BSMMU] and Dhaka Medical College Hospital, Bangladesh. Fifty two, age and gender matched healthy volunteers [42 males and 10 females, mean age of 30 +/- 6 yrs] were also enrolled in this study. Immunoglobulin levels were measured by turbidimetry method using immunoglobulin kits. It was found that the mean serum immunoglobulin concentrations of IgG, IgM and IgA of panic disorder patients were 0.999 +/- 0.26 [g/L], 0.1 +/- 0.028 [g/L] and 0.194 +/- 0.066 [g/L] respectively whereas the values were 1.24 +/- 0.39 [g/L], 0.096 +/- 0.022 [g/L], 0.194 +/- 0.053 [g/L] in healthy volunteers. IgG level in panic disorder patient was found significantly [p <0.05] lower than that of the controls but the change in concentration of IgM and IgA were not significant [p=0.497, p=0.962]. Socioeconomic data reveals that most of the patients were from lower income group and educated. BMI [Mean +/- SD] of the patients [22.62 +/- 3.74 kg/m[2]] and controls [23.74 +/- 2.71 kg/m[2]] were well within the normal range. From correlative analysis it has been found that income has significant effect [p=0.047] on the change of the serum IgG level in panic disorder patient and it was also been justified by the regression analysis [p=0.049]. This finding may play a key role in the diagnosis and treatment of the panic disorder patients. Further studies have been suggested with a large number of populations to confirm these findings