Membrane subproteomic analysis of Mycobacterium bovis bacillus Calmette-Guérin

Mycobacterium bovis bacillus Calmette-Guérin (BCG) vaccine has been known for a long time to prevent tuberculosis (TB) worldwide since 1921. Nonetheless, we know little about BCG membrane proteome. In the present study, we utilized alkaline incubation and Triton X-114-based methods to enrich BCG membrane proteins and subsequently digested them using proteolytic enzyme. The recovered peptides were further separated by 2-D LC and identified by ESI-MS/MS. As a result, total 474 proteins were identified, including 78 integral membrane proteins (IMPs). Notably, 18 BCG IMPs were described for the first time in mycobacterium. Further analysis of the 78 IMPs indicated that the theoretical molecular mass distribution of them ranged from 8.06 to 167.86 kDa and pI scores ranged from 4.40 to 11.60. Functional classification revealed that a large proportion of the identified IMPs (67.9%, 53 out of 78) were involved in cell wall and cell processes functional group. In conclusion, here we reported a comprehensive profile of the BCG membrane subproteome. The present investigation may allow the identification of some valuable vaccine and drug target candidates and thus provide basement for future designing of preventive, diagnostic, and therapeutic strategies against TB.     

Identification of a novel DNA-binding protein from Mycobacterium bovis bacillus Calmette-Guérin

A novel DNA-binding protein expressed (8-10% in total protein) in Mycobacterium bovis bacillus Calmette-Guérin was observed. This protein was designated mycobacterial DNA-binding protein 1 (MDP1). MDP1 recognized bases, sugar moieties, phosphate-backbone on DNA and preferentially bound to DNA guanine and cytosine. In the gel retardation assay, MDP1 preferentially bound to closed circular plasmid DNA than open circular and linear form plasmid DNA and also bound to RNA. MDP1 formed a highly polymerized structure and localized not only in the nucleoid but also at the 50S ribosomal subunits and cell surface. MDP1 was conserved in Mycobacterium thus far examined and the expression was enhanced in stationary growth phases. These results will provide a reasonable basis for further study of the function of MDP1 in living mycobacteria.     

Patterns of care of radiotherapy in México

Aim

This survey is performed to learn about the structure of radiotherapy in México.

Background

Radiation oncology practice is increasing because of the higher incidence of cancer. There is no published data about radiotherapy in México.

Materials and methods

A questionnaire was sent to the 83 registered centers in the database of the Mexican regulatory agency. One out of the 32 states has no radiotherapy. 27 centers from 14 states provided their answers.

Results

829 patients are treated annually with any radiotherapy modality in each center. Two centers have one cobalt machine, 7 have a cobalt and a linac and 10 have more than one linac. Five centers use 2D planning systems, 22 use 3D; 9, conventional simulators; 22, CT based simulation, and 1 center has no simulation. Most of the centers verify beams with films, electronic portal image devices and cone beam CTs are also used. Intensity modulated and image guided radiotherapy are performed in 5 states. Breast, prostate, cervix, lung, rectum and head and neck cancer are the six most common locations. There are 45 public and 38 private centers, 2 dedicated to children. Two gamma knife units, 5 Novalis systems, 1 tomotherapy and 2 cyberknife machines are working. All centers have at least one radiation oncologist, one physicist and one radiotherapist.

Conclusions

Definitive conclusions cannot be drawn from this limited feedback due to a low participation of centers. This survey about radiotherapy in Mexico shows the heterogeneity of equipment as well as medical and technical staff in the whole country.     

Ion-exchange chromatography method for the purification of genomic DNA fraction from Mycobacterium bovis Bacillus Calmette-Guérin

The goal of this study was to provide practical strategies for purifying genomic DNA fraction from Mycobacterium bovis Bacillus Calmette-Guérin (BCG-DNA) by ion-exchange chromatography. A multistep process was developed to purify BCG-DNA. The process consisted of sonication, heating, trypsin digestion, ion-exchange chromatography, gel-filter chromatography, and lyophilization. After ion-exchange chromatography, BCG-DNA was highly purified and possessed potent biological effects. The methods described were efficient and had good reproducibility. Further, this was the first reported chromatography method to purify BCG-DNA.     

Optimization of native agar extraction from Hydropuntia cornea from Yucatán, México

Response surface methodology (RSM) was used to determine optimum extraction conditions for yield, gelling temperature, melting temperature and apparent viscosity of native agar from the red alga Hydropuntia cornea. Two independent variables were selected during extraction, temperature (80, 90 and 100 degrees C) and time (2, 3 and 4 h). The extraction procedure using 100 degrees C and 3 h produced the greatest yield (43.3%), the greatest gelling temperature (32 degrees C) and the greatest melting temperature (78 degrees C). The extraction using 65 degrees C and 4 h produced the optimum statistical apparent viscosity (80.73 cPs), though agar yield under these conditions had low quality for industrial purposes. Temperature and time do not affect melting temperature, but do play an important role in gelling temperature. From the independent variables studied, extraction temperature is most important in producing an increase in yield, and time is the only variable that contributes significantly to increasing viscosity. Most of the physicochemical properties found in the native agar from H. cornea extracted at 100 degrees C/3 h were similar to commercial samples. These findings suggest the feasibility of using this colloid in spreads and soft-texture food products.     

Health Disparities from Economic Burden of Diabetes in Middle-income Countries: Evidence from México

The rapid growth of diabetes in middle-income countries is generating disparities in global health. In this context we conducted a study to quantify the health disparities from the economic burden of diabetes in México. Evaluative research based on a longitudinal design, using cost methodology by instrumentation. For the estimation of epidemiological changes during the 2010–2012 period, several probabilistic models were developed using the Box-Jenkins technique. The financial requirements were obtained from expected case management costs by disease and the application of an econometric adjustment factor to control the effects of inflation. Comparing the economic impact in 2010 versus 2012 (p<0.05), there was a 33% increase in financial requirements. The total amount for diabetes in 2011 (US dollars) was $7.7 billion. It includes $3.4 billion in direct costs and $4.3 in indirect costs. The total direct costs were $.4 billion to the Ministry of Health (SSA), serving the uninsured population; $1.2 to the institutions serving the insured population (Mexican Institute for Social Security–IMSS-, and Institute for Social Security and Services for State Workers-ISSSTE-); $1.8 to users; and $.1 to Private Health Insurance (PHI). If the risk factors and the different health care models remain as they currently are in the analyzed institutions, health disparities in terms of financial implications will have the greatest impact on users’ pockets. In middle-income countries, health disparities generated by the economic burden of diabetes is one of the main reasons for catastrophic health expenditure. Health disparities generated by the economic burden of diabetes suggests the need to design and review the current organization of health systems and the relevance of moving from biomedical models and curative health care to preventive and socio-medical models to meet expected challenges from diseases like diabetes in middle-income countries.     

Biological physics in México

Biological and physical sciences possess a long-standing tradition of cooperativity as separate but related subfields of science. For some time, this cooperativity has been limited by their obvious differences in methods and views. Biological physics has recently experienced a kind of revival (or better a rebirth) due to the growth of molecular research on animate matter. New avenues for research have been opened for both theoretical and experimental physicists. Nevertheless, in order to better travel for such paths, the contemporary biological physicist should be armed with a set of specialized tools and methods but also with a new attitude toward multidisciplinarity. In this review article, we intend to somehow summarize what has been done in the past (in particular, as an example we will take a closer look at the Mexican case), to show some examples of fruitful investigations in the biological physics area and also to set a proposal of new curricula for physics students and professionals interested in applying their science to get a better understanding of the physical basis of biological function.     

Clinical data and molecular analysis of Mycobacterium tuberculosis isolates from drug-resistant tuberculosis patients in Goiás, Brazil

Drug resistance is one of the major concerns regarding tuberculosis (TB) infection worldwide because it hampers control of the disease. Understanding the underlying mechanisms responsible for drug resistance development is of the highest importance. To investigate clinical data from drug-resistant TB patients at the Tropical Diseases Hospital, Goiás (GO), Brazil and to evaluate the molecular basis of rifampin (R) and isoniazid (H) resistance in Mycobacterium tuberculosis. Drug susceptibility testing was performed on 124 isolates from 100 patients and 24 isolates displayed resistance to R and/or H. Molecular analysis of drug resistance was performed by partial sequencing of the rpoB and katGgenes and analysis of the inhA promoter region. Similarity analysis of isolates was performed by 15 loci mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing. The molecular basis of drug resistance among the 24 isolates from 16 patients was confirmed in 18 isolates. Different susceptibility profiles among the isolates from the same individual were observed in five patients; using MIRU-VNTR, we have shown that those isolates were not genetically identical, with differences in one to three loci within the 15 analysed loci. Drug-resistant TB in GO is caused by M. tuberculosis strains with mutations in previously described sites of known genes and some patients harbour a mixed phenotype infection as a consequence of a single infective event; however, further and broader investigations are needed to support our findings.     

Acetobacter diazotrophicus,an indoleacetic acid producing bacterium isolated from sugarcane cultivars of México

Thirteen cane cultivars grown on fields in México were sampled to assess the occurrence of Acetobacter diazotrophicus, a recently identified N₂-fixing bacterium. Results showed that the isolation frequencies extended over a broad range (1.1 to 67%), likely to be related to the nitrogen fertilization level. The lowest isolation frequencies (1.1 to 2.5%) were obtained from plants growing at high nitrogen doses (275–300 kg ha^-1) and the highest values (10–67%) from plants cultivated with 120 kg N ha^-1. All eighteen strains of A. diazotrophicus produced indoleacetic acid (IAA) in defined culture medium. Estimates obtained from HPLC analyses revealed that A. diazotrophicus strains produced from 0.14 to 2.42 g IAA mL^-1 in culture medium. Considering that A. diazotrophicus is found within the plant tissue, the biosynthesis of IAA suggests that the bacteria could promote rooting and improve sugarcane growth by direct effects on metabolic processes, in addition to their role in N₂ fixation.     

Ichthyofauna from wetlands of San Pedro, Balancán, Tabasco, México

San Pedro River's wetlands sustain trophic nets in the fluvial system, due to the high habitat availability, and space and temporal variations. In order to describe the relationship between environmental parameters and ichthyofauna, this study evaluated fish assemblages composition, distribution, abundance, density, biomass, richness species, diversity and equitability in the wetlands. Sampling considered three different sites and climatic seasons (dry, rainy and cold fronts). The physical and chemical parameters considered were dissolved oxygen, temperature, pH, water transparency and the depth. Fishes were caught with a shrimp net, after six minutes towings and were identified afterwards. A total of 1 049 organisms of 25 fish species were caught, two of which were exotic species: Oreochromis niloticus and Parachromis managuense. A total of 23 species were found at site I (with the highest density 0.23 ind./m2), 17 at site II (0.23 ind./m2) and 14 at site III (0.12 ind./m2). The dry season had the highest species number with 21 species, followed by the rainy season with 17 species, and the cold season with five species. Similarly, the highest biomass (8.30 g/m2) was found in dry season, followed by the rainy (2.16 g/m2) and the cold seasons (0.03 g/m2). Considering seasons, highest density was found during the dry (0.436 ind./m2), followed by the rainy (0.213 ind./m2) and the cold (0.023 ind./m2) seasons. The dominant density species during the study period, according to the quadrants graphic of Olmstead-Tukey were: Petenia splendida, Vieja heterospila, Vieja synspila, Dorosoma petenense and Astyanax aeneus. There were significant differences in the species richness among sites. Temperature, depth and transparency showed differences among the seasons. The canonical correspondence analysis indicated that fish distribution was governed by environmental parameters during all seasons. In terms of fish abundance and composition, environmental parameters play an important role showing spatial and temporal differences in the ecosystem, this could be explained with the fact that most of young fishes have a movement behavior to the wetlands, searching refuge and feed during the dry season. Considering the diversity indexes variation, it may be concluded that San Pedro River's wetlands correspond to a system where the ichthyofauna composition fluctuates spatial and seasonally.     

Perkinsus marinus in pleasure oyster Crassostrea corteziensis from Nayarit, Pacific coast of México

Culture of the pleasure oyster Crassostrea corteziensis is emerging as an alternative to the Pacific oyster (Crassostrea gigas) for oyster producers, who face severe mortalities since 1997 in Northwest México. For determining the health status of this species, we conducted a histopathological analysis of cultured populations from two estuaries in the Pacific coast of México. Macroscopical analysis revealed animals with transparent and retracted mantle. Histopathological analysis of these specimens showed tissue alterations and parasitic forms consistent with Perkinsus sp. infection. Stages of the parasite identified included tomont and trophozoites with an eccentric vacuole characteristic of Perkinsus spp. Pieces of tissues of infected oysters were incubated in Fluid Thioglycollate Medium (FTM) resulting in blue–black hypnospores after incubation. The identity of the parasite was confirmed by species specific PCR-based assay in DNA samples from oysters, tissue fractions from FTM cultures, and deparaffined samples with Perkinsus-like parasite detected by histology. Sequencing of positive amplified fragments (307 bp) showed a sequence similar to Perkinsus marinus strain TXsc NTS ribosomal RNA gene (100% coverage and 98% identity, GenBank Accession No. AF497479.1) and to P. marinus, Genomic DNA, (100% coverage and 97% identity, GenBank Accession No. S78416.1). The prevalence of P. marinus varied from 1 to 5% in Boca del Camichín and from 1 to 6% in Pozo Chino. In general, the intensity of infection was moderate. The infection was observed in oysters from 31 to 110 mm of shell length. This is the first record of P. marinus in oysters from the North America Pacific coast and the first record in C. corteziensis. The origin of this parasite in the area is unknown, but it may be associated to introductions of Crassostrea virginica from the East coast of United States of America or Gulf of México.     

Gastropods and bivalves of commercial interest from the continental shelf of Jalisco and Colima,México

The distribution and abundance with respect to depth and type of substratum of 20 species of gastropods and four species of bivalves of economic importance were examined in the continental shelf of Jalisco and Colima, México. These species were taken with net trawls at depths from 24 to 83 m in August, 1988. Most individuals and species of gastropods were collected in stations with sandy silt substratum. Bivalves were collected in sandy silt and medium sand substrata. The six most abundant species represented 81.2% of all gastropods and bivalves collected. These species are: Cantharus pallidus, Fusinus dupetittouarsi, Ficus ventricosa, Hexaplex brassica, Harpa conoidalis and Arca pacifica.     

Population biology patterns of commercially important mollusks in México

The need to propose recommendations for the management of over 80 species of bivalves and gastropod mollusks exploited commercially in Mexico, led to look for trends on the population and reproductive biology in relation to prevailing environmental conditions of the habitats where they are exploited. The reproductive cycle, growth parameters for the von Bertalanffy equation, mortality and recruitment of 18 populations of 14 species from 13 localities are compared and related to ambient temperature, precipitation, evaporation, geomorphology, tides, and water salinity and temperature. Localities were classified as influenced by landmasses or with marine influence, with a desert or tropical humid climate. With restricted or continuous communication to oceanic waters and with or without freshwater runoff. The reproductive cycles were classified in four groups in relation to intensity and duration of the spawning season: 1) limited to one annual spawning, 2) two or more defined spawning periods, 3) two or more extended spawning periods, and 4) continuous low intensity spawning. And three groups in relation to gonad recovery, and duration and intensity of gametogenesis: 1) post spawn and rest stages absent or restricted, 2) fast gametogenesis and a clear mature stage. and 3) extended gametogenesis and limited maturity stages. The population parameters were classified in relation to age structure and number of cohorts, intensity and duration of recruitment, and growth rates as expressed by infinity and K. In relation to their life cycle four types were found: 1) population represented by only one cohort, at least during part of the year, 2) with two or more cohorts at any time, 3) longevity under five years, and 4) longevity over live years. In relation to recruitment: 1) one recruitment period restricted to a short season, 2) two or more recruitment periods, 3) constant recruitment with one or more peaks, and 4) constant recruitment without periods of dominance.     

A new glossiphoniid leech from Catemaco Lake, Veracruz, México

Haementeria acuecueyetzin n. sp. from Catemaco Lake, Veracruz, Mexico, is described based on the examination of 6 specimens. This new hematophagous leech species resembles other members of the genus in the number and position of the eyespots, number of compact salivary glands, and in the presence of 2 pairs of spheroidal mycetomes, but it is distinguished from the other species by having 6 rows of longitudinal smooth white papillae in the dorsal surface and numerous tubercles in dorsal and ventral surfaces. This new species represents the third species of Haementeria in the Northern Hemisphere of the Americas.     

Dynamics of a parasite assemblage of the Vermilion Rockfish Sebastes miniatus from northwestern Baja California,México

A parasite assemblage of Sebastes miniatus from northwestern Baja California, México, was composed of a total of 12 species: five ectoparasites (two monogeneans and three parasitic copepods) and seven endoparasites (two digeneans, one cestode, three nematodes, and one acanthocephala). Five of these parasites constituted new genera records to the genus Sebastes, and nine were new geographic records. The most abundant species were the endoparasites Parabothriocephalus sagitticeps, Hysterothylacium sp., and Anisakis sp., and the specific richness ranged from 1 to 8 parasite species per host. The most important parasite species in terms of prevalence were Microcotyle sebastis (93 %) and Anisakis sp. (92 %). The mean abundance of parasites found in S. miniatus showed significant variations over the year, with maximum values (31.7 individuals/host) in August, and minimum (0.39 individuals/host) in February. P. sagitticeps showed the highest mean intensity of infection (190.4 parasites/host), followed by Anisakis sp. (127.2 parasites/host) and Hysterothylacium sp. (46.6 parasites/host). The presence of larval stages of the nematodes Anisakis, Pseudoterranova, and Hysterothylacium is particularly important due to their high abundance and prevalence and because they may represent a human health risk (anisakiasis). Rockfishes (family Scorpaenidae) of the genus Sebastes constitute one of the most important groundfish resources in the American and Mexican northern Pacific Ocean, both for recreational and for the commercial fisheries of California and Baja California. These rockfish species makes up a substantial part of the Mexican cuisine.     

Growth inhibition of HeLa cell by internalization of Mycobacterium bovis Bacillus Calmette-Guérin (BCG) Tokyo

Background  

Intravesical BCG immunotherapy is effective for preventing recurrence and progression in none muscle-invasive bladder cancer but the dosing schedule and duration of treatment remain empirical. The mechanisms by which intravesical BCG treatment mediates antitumor activity are currently poorly understood.     

Susceptibility to mercurials of clinical Pseudomonas aeruginosa isolated in México

Susceptibility to inorganic mercuric ions and to organomercurials of 237 Pseudomonas aeruginosa clinical strains isolated in Mexico was determined by agar dilution tests. Resistant strains fell into two classes: i) narrow-spectrum resistant strains (27% of total isolates) resistant only to mercuric ions and to merbromin, and most grouped in pyocin type 1; and ii) broad-spectrum resistant strains (5%) with additional resistances to thimerosal, phenylmercury, methylmercury and p-hydroxymercuribenzoate, that belonged mostly to pyocin type 10. Mercurial resistant isolates showed a higher proportion of resistance to antibiotics and metals than did mercurial sensitive isolates, and broad-spectrum resistant strains had the highest frequency of resistance to antibiotics and to tellurite and arsenate.     

The Ecitoninae ants (Hymenoptera: Formicidae) from Morelos, México

To produce an inventory of the Ecitoninae ants from Morelos State (south central Mexico), we used direct capture and pit-fall traps. Occasionally, males were also collected near artificial light sources. Fifteen species were found: Labidus coecus, L. praedator s. str, Neivamyrmex agilis, N. cornutus, N fallax, N. graciellae, N. impudens, N. macropterus, N. melanocephalus, N. nigrescens, N. opacithorax, N. pauxilus, N. sumichrasti, N. swainsoni and Nomamyrmex esenbecki mordax. Twelve of these species are new records for the state. The most abundant species were L. coecus, N. melanocephalus, N. nigrescens and N. esenbecki mordax. The distribution of the species in the state, flight dates of males for some species, and a key for the identification of workers and males are included. One additional species (N. fuscipennis), collected by other authors in Morelos but not by us, is also included in the key.