Utilizing bioinformatics to detect genetic similarities between African honey bee subspecies

Various honey bees, especially subspecies Apis mellifera, occur in Africa and are distribute across the continent. The genetic relationships and identical genetic characteristics between honey bee subspecies in Africa (African bee subspecies) have not been widely investigated using sequence analysis. On the other hand, bioinformatics are developed rapidly and have diverse applications. It is anticipated that bioinformatics can show the genetic relationships and similarities among subspecies. These points have high importance, especially subspecies with identical genetic characteristics can be infected with the same group of pathogens, which have implications on honey bee health. In this study, the mitochondrial DNA sequences of four African subspecies and Africanized bees were subjected to the analyses of base composition, phylogeny, shared gene clusters, enzymatic digestion, and open reading frames. High identical base composition was detected in the studied subspecies, and high identical results from all tests were found between A. m. scutellata and A. m. capensis followed by A. m. intermissa and A. m. monticola. The least genetic relationships were found between A. m. lamarckii and the other subspecies. This study presents insights into the genetic aspects of African bee subspecies and highlights similarity and dissimilarity aspects. Also, Africanized honey bees derived from A. m. scutellata showed high genetic similarities to other African bees, especially A. m.capensis. Additionally, specific primers to identify these subspecies were designed and tested.

Implementación hardware del algoritmo de Needleman-Wunsch modificado usando una arquitectura paralela / Hardware implementation of modified Needleman-Wunsch algorithm using a parallel architecture / Implementação de hardware do algoritmo modificado Needleman-Wunsch usando uma arquitetura paralela

Resumen Este artículo presenta el diseño de un procesador para el alineamiento global de pares de cadenas de ADN. El principal bloque funcional del procesador es un arreglo paralelo de dos dimensiones que permite realizar cálculos simultáneos, reduciendo el tiempo de procesamiento con respecto a la implementación software. En este trabajo, el hardware diseñado lleva a cabo la alineación de dos secuencias de más de 400 nucleótidos correspondientes a la proteína de transición 1 (Tnp1) de la rata parda y el ratón común. El algoritmo implementado es k-band, una modificación del algoritmo de alineamiento global Needleman-Wunsch, donde se realizan únicamente cálculos sobre las diagonales principales de la matriz, formando una banda que puede ser de un tamaño variable. Se realizan simulaciones del diseño propuesto usando bandas de K=2, 4, 6, 8 y 10.

Abstract This paper proposes a DNA sequence pair global alignment processor design. The processor main functional block is a two-dimensional parallel array that allows simultaneous computations, reducing the processing time compared to the software implementation. In this work, the designed hardware performs over 400 nucleotides sequence alignment corresponding to the Mus musculus transition protein 1 (Tnp1), mRNA and the Rattus norvegicus transition protein 1 (Tnp1), mRNA. The implemented algorithm is k-band, a Needleman-Wunsch global alignment algorithm modification, where calculations are made only on the matrix diagonals, establishing a band that can be of a variable size. Simulations of the proposed design are performed using K = 2, 4, 6, 8 and 10 bands.

Resumo Este artigo propõe um projeto de processador de alinhamento global de pares de cadeia de DNA. O bloco funcional principal do processador é uma matriz bidimensional paralela que permite cálculos simultâneos, reduzindo o tempo de processamento para a implementação do software. Neste trabalho, o hardware projetado executa o alinhamento de duas sequências de mais de 400 nucleótidos correspondente à proteína de transição 1 (Tnp1) do rato marrom e ao mouse comum. O algoritmo implementado é k-band, uma modificação do algoritmo de alinhamento global Needleman-Wunsch, onde os cálculos são feitos apenas nas diagonais da matriz, estabelecendo uma banda que pode ser de tamanho variável. As simulações do projeto proposto são realizadas usando K = 2, 4, 6, 8 e 10 bandas.

Indole acetic acid production by fluorescent Pseudomonas spp. from the rhizosphere of Plectranthus amboinicus (Lour.) Spreng. and their variation in extragenic repetitive DNA sequences.

Fluorescent Pseudomonas (FP) is a heterogenous group of growth promoting rhizobacteria that regulate plant growth by releasing secondary metabolic compounds viz., indole acetic acid (IAA), siderophores, ammonia and hydrogen cyanide. In the present study, IAA producing FPs from the rhizosphere of Plectranthus amboinicus were characterized morphologically, biochemically and at the molecular level. Molecular identification of the isolates were carried out using Pseudomonas specific primers. The effect of varying time (24, 48, 72 and 96 h), Trp concentrations (100, 200, 300, 400 and 500 µg.ml-1), temperature (10, 26, 37 and 50±2 °C) and pH (6, 7 and 8) on IAA production by 10 best isolates were studied. Results showed higher IAA production at 72 h incubation, at 300 µg.ml-1 Trp concentration, temperature 26±2 °C and pH 7. TLC with acidified ethyl acetate extract showed that the IAA produced has a similar Rf value to that of the standard IAA. Results of TLC were confirmed by HPLC analysis. Genetic diversity of the isolates was also studied using 40 RAPD and 4 Rep primers. Genetic diversity parameters such as dominance, Shannon index and Simpson index were calculated. Out of 40 RAPD primers tested, 9 (2 OP-D series and 7 OP-E series) were shortlisted for further analysis. Studies using RAPD, ERIC, BOX, REP and GTG5 primers revealed that isolates exhibit significant diversity in repetitive DNA sequences irrespective of the rhizosphere.

Morphology of the larvae, male genitalia and DNA sequences of Anopheles (Kerteszia) pholidotus (Diptera: Culicidae) from Colombia

Since 1984, Anopheles (Kerteszia) lepidotus has been considered a mosquito species that is involved in the transmission of malaria in Colombia, after having been incriminated as such with epidemiological evidence from a malaria outbreak in Cunday-Villarrica, Tolima. Subsequent morphological analyses of females captured in the same place and at the time of the outbreak showed that the species responsible for the transmission was not An. lepidotus, but rather Anopheles pholidotus. However, the associated morphological stages and DNA sequences of An. pholidotus from the foci of Cunday-Villarrica had not been analysed. Using samples that were caught recently from the outbreak region, the purpose of this study was to provide updated and additional information by analysing the morphology of female mosquitoes, the genitalia of male mosquitoes and fourth instar larvae of An. pholidotus, which was confirmed with DNA sequences of cytochrome oxidase I and rDNA internal transcribed spacer. A total of 1,596 adult females were collected in addition to 37 larval collections in bromeliads. Furthermore, 141 adult females, which were captured from the same area in the years 1981-1982, were analysed morphologically. Ninety-five DNA sequences were analysed for this study. Morphological and molecular analyses showed that the species present in this region corresponds to An. pholidotus. Given the absence of An. lepidotus, even in recent years, we consider that the species of mosquitoes that was previously incriminated as the malaria vector during the outbreak was indeed An. pholidotus, thus ending the controversy.

Effects of Immunostimulatory CpG-Oligodeoxynucleotides of Bronchial Asthma in Rat / 결핵

BACKGROUND AND OBJECT: Immunostimulatory CpG-oligodeoxynucleotides (ISS CpG-ODN) up-regulate the TH1-type immune response and down-regulate the TH2-type response. This study was performed to investigate the immune response changes resulting from ISS CpG-ODN on bronchial hyperrestponsiveness, eosinophilic inflammation and mucus hypersecretion in rat asthma. MATERIALS AND METHODS: 10 normal controls(NC) and 26 asthmatic rats, which were generated by ovallbumin(OVA) sensitization and challenge, were studied. The asthmatic rats were randomized into 11 asthma controls(AC) and 15 in the asthma-CpG treatment group(CpG). The CpG group was administered ISS CpG-ODN intramuscularly and the AC group was administered a placebo(0.9% NaCl)on day 15 and 20. After CpG-ODN or placebo administration, we measured the IFN-(TH1-type cytokine) and IL-4(TH2-type cytokine) levels in the bronchoalveolar lavage fluid(BALF), the specific airway resistance(sRaw), eosinophilic fraction in BALF, eosinophilic infiltration, goblet cell dysplasia and MUC5AC gene expression in the lung tissue. RESULTS: In the BALF of the CpG group, the IFN-γ concentration was significantly high and the IL-4 concentration was significantly low when compared with the AC group. Both the sRaw and eosinophilic fraction, and infiltration into the BALF and lung tissue significantly lower in the CpG group when compared with the AC group. However, little difference in goblet cell dysplasia and MUC5Ac gene expression was observed between the CpG group and the Ac group. CONCLUSION: ISS CpG-ODN decreases bronchial hyperresponsiveness and eosinophilic inflammation in the rat asthma model through the up-regulation of the TH1-type immune response with the down-regulation of the TH2-type response. However, the effect of these immune response changes on mucus hypersecretion was is not remarkable in this study.

Analysis of sequences of two different classes of Kinetoplast DNA minicircles of a Leishmania Spp.

We have determined the nucleotide sequences of the minicircles representing a major (pLURkE3) and a minor (pLURkH13) class populations from the kinetoplast DNA of Leishmania strain UR6. These minicircles have sequence organization similar to other kinetoplastid parasites, however, they have some unique structural features. These features include the following: (i) imperfect inverted repeat in the variable regions, similar to the conserved sequence elements of guide RNA genes in African trypanosomes, (ii) tandem and non-tandem direct repeats of 8 bp or longer scattered throughout the minicircles, (iii) non uniform strand distribution of bases throughout the minicircles and (iv) high TG content, more than half of the molecules being extremely (T + G) versus (A + C) strand biased. The heterogeneity of minicircle sequences in the variable regions may be exploited in developing recombinant DNA based diagnostic probes for detection and classification of Leishmania species.

Computer simulation of the interaction of a3helix of λ Cro protein with DNA and origin of sequence specific recognition.

The conformation of the a3 helix of Cro protein (residues 27–36) of bacteriophage λ is optimised by the damped least square minimization technique, with the steric constraint that Ca atom positions should match the crystallographic data available to date. On the basis of minimization of total interaction and conformation energy, models for complexes of this peptide sequence with heptanucleotide duplexes from native and altered OR3 operator are obtained in the major groove of Β DNA. Analysis of the energetics for 3 sequences of the DNA show that binding strength is derived mainly from the interaction of side chains of the peptide with DNA. Sequence specificity (maximum difference in binding energy for different DNA sequences) is due to hydrogen bonding interaction. A small amount of sequence specificity is derived from non-bonded interaction also. Stereochemical aspects of peptide DNA interaction and their role in DNA recognition are discussed in this paper.