Exploring the non-coding regions in the mtDNA of some honey bee species and subspecies

The sequence of the DNA contains coding and non-coding regions. The role of the non-coding regions is not known and is hypothesized to maintain the structure of the DNA. This study aimed to investigate the structure of the non-coding sequences in honey bees utilizing bioinformatics. The non-coding sequences of the mtDNA of three honey bee species Apis dorosata, Apis florea, Apis cerana, and ten subspecies of Apis mellifera were investigated. Different techniques were utilized to explore the non-coding regions of these bees including sequence analysis, phylogenetic relationships, enzymatic digestion, and statistical tests. Variations in size and sequences of nucleotides were detected in the studied species and subspecies, but with the same nucleotide abundance (i.e. nucleotides A were more than T and nucleotides G were less than C). The phylogenetic tree based on the non-coding regions was partially similar to the known phylogenetic relationships between these bees. The enzymatic digestion using four restriction enzymes confirmed the results of the phylogenetic relationships. The statistical analysis based on numerical codes for nucleotides showed the absence of significant variations between the studied bees in their sequences in a similar way to results of neutrality tests. This study suggests that the non-coding regions have the same functional role in all the studied bees regardless of the number of nucleotides, and not just to maintain the structure of the DNA. This is approximately the first study to shade lights on the non-coding regions of the mtDNA of honey bees.     

Recombination and population structure in Salmonella enterica

Salmonella enterica is a bacterial pathogen that causes enteric fever and gastroenteritis in humans and animals. Although its population structure was long described as clonal, based on high linkage disequilibrium between loci typed by enzyme electrophoresis, recent examination of gene sequences has revealed that recombination plays an important evolutionary role. We sequenced around 10% of the core genome of 114 isolates of enterica using a resequencing microarray. Application of two different analysis methods (Structure and ClonalFrame) to our genomic data allowed us to define five clear lineages within S. enterica subspecies enterica, one of which is five times older than the other four and two thirds of the age of the whole subspecies. We show that some of these lineages display more evidence of recombination than others. We also demonstrate that some level of sexual isolation exists between the lineages, so that recombination has occurred predominantly between members of the same lineage. This pattern of recombination is compatible with expectations from the previously described ecological structuring of the enterica population as well as mechanistic barriers to recombination observed in laboratory experiments. In spite of their relatively low level of genetic differentiation, these lineages might therefore represent incipient species.     

Molecular characterization of secretory proteins Rv3619c and Rv3620c from Mycobacterium tuberculosis H37Rv

Rv3619c and Rv3620c are the secretory, antigenic proteins of the ESAT-6/CFP-10 family of Mycobacterium tuberculosis H37Rv. In this article, we show that Rv3619c interacts with Rv3620c to form a 1 : 1 heterodimeric complex with a dissociation constant (K(d)) of 4.8 × 10(-7) M. The thermal unfolding of the heterodimer was completely reversible, with a T(m) of 48 °C. The comparative thermodynamics and thermal unfolding analysis of the Rv3619c-Rv3620c dimer, the ESAT-6-CFP-10 dimer and another ESAT family heterodimer, Rv0287-Rv0288, revealed that the binding strength and stability of Rv3619c-Rv3620c are relatively lower than those of the other two pairs. Molecular modeling and docking studies predict the structure of Rv3619c-Rv3620c to be similar to that of ESAT-6-CFP-10. Spectroscopic studies revealed that, in an acidic environment, Rv3619c and Rv3620c lose their secondary structure and interact weakly to form a complex with a lower helical content, indicating that Rv3619c-Rv3620c is destabilized at low pH. These results, combined with those of previous studies, suggest that unfolding of the proteins is required for dissociation of the complex and membrane binding. In the presence of membrane mimetics, the α-helical contents of Rv3619c and Rv3620 increased by 42% and 35%, respectively. In mice, the immune response against Rv3619c protein is characterized by increased levels of interferon-γ, interleukin-12 and IgG(2a) , indicating a dominant Th1 response, which is mandatory for protection against mycobacterial infection. This study therefore emphasizes the potential of Rv3619c as a subunit vaccine candidate.     

Arsenic accumulation in Ocimum spp. and its effect on growth and oil constituents

A comparative evaluation of As accumulation and subsequent effects upon exposure to arsenite [As(III)] was performed in three species of Ocimum. Plants accumulated high amount of As (μg g^?1 dry weight; dw) (662 in O. tenuiflorum, 764 in O. basilicum and 831 in O. gratissimum at 100 μM As(III) after 10 days) with the order of accumulation being roots > stem > leaves. A significant reduction in plant height and biomass was observed. However, essential oil yield and major oil constituents, such as eugenol, methyl chevicol, and linalool, increased at lower As(III) concentrations [mostly up to 25 μM As(III)] in all three species. Positively, no detectable amount of As was found in oil of any species. The study proposes that Ocimum may be used as a phytoremediator and at the same time as a source of essential oils under proper regulation.     

Biotransformation of 17α-ethynyl substituted steroidal drugs with microbial and plant cell cultures: A review

Structural modification of steroids through whole-cell biocatalysis is an invaluable procedure for the production of active pharmaceutical ingredients (APIs) and key intermediates. Modifications could be carried out with regio- and stereospecificity at positions hardly available for chemical agents. Much attention has been focused recently on the biotransformation of 17α-ethynyl substituted steroidal drugs using fungi, bacteria and plant cell cultures in order to obtained novel biologically active compounds with diverse structure features. Present article includes studies on biotransformation on 17α-ethynyl substituted steroidal drugs using microorganisms and plant cell cultures. Various experimental and structural elucidation methods used in biotransformational processes are also highlighted.     

Molecular characterization of the modular chitin binding protein Cbp50 from <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> serovar <Emphasis Type="Italic">konkukian</Emphasis>

Bacillus thuringiensis is an insecticidal bacterium whose chitinolytic system may be exploited to improve the insecticidal system of Bt-crops. A nucleotide fragment of 1368 bp from B. thuringiensis serovar konkukian S4, containing the complete coding sequence of the chitin binding protein Cbp50, was cloned and sequenced. Analyses have shown the protein to contain a modular structure consisting of an N-terminal CBM33 domain, two copies of a fibronectin-like domain and a C-terminal chitin binding domain classified as CBM5. The Cbp50 protein was heterologously expressed in Escherichia coli, purified and assessed for chitin binding activity. A deletion mutant (CBD-N; containing only the N-terminal CBM33 domain) of Cbp50 was produced to determine the role of C-terminal domains in the binding activity of the protein. The full-length Cbp50 was shown to bind β-chitin most efficiently followed by α-chitin, colloidal chitin and cellulose. The polysaccharide binding activity of CBD-N was drastically decreased. The data demonstrate that both the N-terminal and C-terminal domains of Cbp50 are essential for the efficient binding of chitin. The purified Cbp50 showed antifungal activity against the phytopathogenic fungus Fusarium oxysporum and the opportunistic human pathogen Aspergillus niger. This is the first report of a modular chitin binding protein in bacteria.     

Cytotoxicity and mechanical behavior of chitin-bentonite clay based polyurethane bio-nanocomposites

Chitin based polyurethane bio-nanocomposites (PUBNC) were prepared using chitin, Delite^® HPS bentonite nanoclay enriched in montmorillonite (MMT), 4,4′-diphenylmethane diisocyanate (MDI) and polycaprolactone polyol CAPA 231 (3000 g/mol⁻¹). The prepolymers having different concentration of Delite HPS bentonite nanoclay were extended with 2 moles of chitin. The structures of the resulted polymers were determined by FT-IR technique. The effect of nanoclay contents on mechanical properties and in vitro biocompatibility was investigated. The mechanical properties of the synthesized materials were improved with increase in the Delite HPS^® bentonite nanoclay contents. Optimum mechanical properties were obtained from the PU bio-nanocomposite samples having 4% Delite HPS^® bentonite nanoclay. The results revealed that the final PU bio-nanocomposite having 2% Delite HPS^® bentonite nanoclay contents is ideal contenders for surgical threads with on going investigations into their in vitro biocompatibility, non-toxicity, and mechanical properties.     

Effect of Mineral Fortification on Plasma Biochemical Profile in Rats

This study aimed at investigating the changes in biochemical profile of male rats following 8 weeks administration of different concentration of elemental iron, sodium iron ethylenediaminetetraacetate (NaFeEDTA), zinc sulfate (ZnSO₄), and zinc oxide (ZnO) in whole wheat flour. Eight groups comprising five rats each were fed fortified whole wheat flour in the form of baked pallets, while one group served as control. Concentration of total cholesterol, high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), triglycerides, total proteins, albumin, globulin, plasma glucose, and blood urea nitrogen were assayed. Supplementing mineral-fortified diet to male rats did not indicate any significant (p ≤ 0.05) effect on total cholesterol concentration. Diets containing NaFeEDTA alone increased HDL-C and decreased LDL-C; however, the differences remained non significant. Likewise, plasma triglycerides content of male rats remained unchanged on feeding fortified diets. Diets containing iron as NaFeEDTA and elemental iron exerted little effect on total protein concentration in the plasma of rats. Plasma glucose and blood urea nitrogen levels did not exhibit any significant change as a result of ingesting mineral supplemented diets. The study concludes that the forms of fortificants and the fortification levels used in the current study are undamaging for lipid profile, renal function, and glucose levels in rats, suggesting that these may be safely used in wheat flour to combat iron and zinc deficiency in vulnerable groups.