Mobility and accessibility of Zn, Pb, and As in abandoned mine tailings of northwestern Mexico.

Generation, storage, and management of waste coming from industrial processes are a growing worldwide problem. One of the main contributors is the mining industry, in particular tailings generated by historical mining, which are barely maintained, especially in developing countries. Assessing the impact of a mining site to surrounding soils and ecosystems can be complex, especially when determining mobility and accessibility of the contaminants is required to perform ecological and human health risk assessment. As an effort to obtain information regarding mobility and accessibility of some potentially toxic elements (Zn, Pb, and As) from an historical mining site of northwestern Mexico, the abandoned mine tailings of San Felipe de Jesús in central Sonora and adjacent agricultural soils were investigated. Mobility and accessibility were assessed by means of sequential extraction procedures and using simulated physiological media. Additionally, an assessment of accidental oral intake was calculated considering the bioaccessible fractions. Results show that higher concentrations of contaminants were found in sulfide-rich tailings (Zn = 92,540; Pb = 21,288; As = 19,740 mg kg-1) compared with oxide-rich tailings (Zn = 43,240; Pb = 14,763; As = 13,401 mg kg-1). Concentrations in agricultural soils were on average Zn = 4755, Pb = 2840, and As = 103 mg kg-1. Zinc was mainly recovered from labile fractions in oxide-rich tailings (~ 60%) and in a lower amount from sulfide-rich tailings (~ 30%). Pb and As were mainly associated with residual fractions (80-95%) in both types of tailings. The percentage of mobile fractions (sum of water-soluble, exchangeable, and bound to carbonate fractions) in agricultural soils was as follows: Zn ~ 60%, Pb ~ 15%, and As ~ 70%. Regarding the phytoaccessible fraction, the studied elements in mine tailings and agricultural soil samples exceeded the threshold limits, except for As in agricultural soils. According to data obtained, toxic effects were also calculated. As for daily oral intake for non-carcinogenic effects in adults and children, only Pb and As exceeded reference dose values, especially in children exposed to sulfide-rich tailings and agricultural soils. Regarding carcinogenic effects of Pb and As, most of the samples were above acceptable risk values.

Human papillomavirus DNA and protein in tissue samples of oesophageal cancer, Barrett's oesophagus and oesophagitis.

BACKGROUND: The risk of presenting with oesophageal cancer is associated with Barrett's oesophagus, with a higher prevalence in some Asian and African countries. Human papillomavirus (HPV) DNA has been identified in oesophageal carcinomas, which share common features with cervical cancers and originate in stratified epithelium. MATERIALS AND METHODS: Sixty-eight paraffin-embedded tissue biopsies were selected from Mexican patients: 17 from oesophageal cancers, 28 from cases of Barrett's oesophagus and 23 from cases of oesophagitis. HPV protein was detected immunohistochemically and the presence and types of HPV DNA were assessed by polymerase chain reaction. RESULTS: HPV DNA-positive results were found in 26% of samples of oesophagitis, 96% of samples of Barrett's oesophagus and 88% of samples of oesophageal cancers. HPV viral types 6 and 11 were prevalent. HPV protein was detected in 41 samples (60%). CONCLUSION: Mexico has a high prevalence of HPV in premalignant and malignant oesophageal diseases compared with other countries.

Molecular and functional characterization of adenylate kinase 2 gene from Leishmania donovani.

ATP-regenerating enzymes may have an important role in maintaining ATP levels in mitochondria-like kinetoplast organelle and glycosomes in parasitic protozoa. Adenylate kinase (AK) (ATP:AMP phosphotransferase) catalyses the reversible transfer of the gamma-phosphate group from ATP to AMP, releasing two molecules of ADP. This study describes cloning and functional characterization of the gene encoding AK2 from a genomic library of Leishmania donovani and also its expression in leishmania promastigote cultures. AK2 was localized on an approximately 1.9-Mb chromosomal band as a single copy gene. L. donovani AK2 gene is expressed as a single 1.9-kb mRNA transcript that is developmentally regulated and accumulated during the early log phase. The overexpression of L. donovani AKgene in Escherichia coli yielded a 26-kDa polypeptide that could be refolded to a functional protein with AK activity. The recombinant protein was purified to apparent homogeneity. Kinetic analysis of purified L. donovani AK showed hyperbolic behaviour for both ATP and AMP, with Km values of 104 and 74 microM, respectively. The maximum enzyme activity (Vmax) was 0.18 micromol.min(-1).mg(-1) protein. P1,P5-(bis adenosine)-5'-pentaphosphate (Ap5A), the specific inhibitor of AK, competitively inhibited activity of the recombinant enzymes with estimated Ki values of 190 nM and 160 nM for ATP and AMP, respectively. Ap5A also inhibited the growth of L. donovani promastigotes in vitro which could be only partially reversed by the addition of ADP. Thus, presence of a highly regulated AK2, which may have role in maintenance of ADP/ATP levels in L. donovani, has been demonstrated.

Identification of a gene in Leishmania infantum encoding a protein that contains a SP-RING/MIZ zinc finger domain.

The SP-RING or Miz zinc finger domain that is related to the classical RING-finger motif, defines a class of proteins that can act as E3-like factors in the pathway of small ubiquitin-related modifier (SUMO) conjugation. This family includes the mammalian protein inhibitor of activated STAT (PIAS) proteins and related proteins from lower eukaryotes. Here we report the existence of a gene in Leishmania infantum, present as two identical copies placed upstream of each MAT2 gene copy, and transcribed as a single approximately 2.2 kb mRNA both in the logarithmic and stationary phases of the promastigote stage. This gene encodes a 47 kDa protein that has been named LORIEN. LORIEN is circumscribed to the cell periphery and it is antigenic during L. infantum infection of dogs and hamsters. Strikingly, this novel protein contains a highly conserved SP-RING/Miz zinc finger domain, raising the possibility that a SUMO or ubiquitin-like system may exist in this microorganism.

[Biochemical and histologic changes in patients with chronic hepatitis C under treatment with interferon. a 5-year cohort study]. / Cambios bioquímicos e histológicos en pacientes con hepatitis crónica por virus C en tratamiento con interferón. Estudio de una cohorte a cinco años.

Since 1989, some schemes of treatment for Chronic hepatitis C (CHC) have been investigated, including alpha and beta interferon (IFN), ribavirin, and amantadine either alone or in combination. These studies have been performed in other countries. In our study, we investigated the outcome of Mexican patients with CHC treated with IFN. Twenty-six patients with five years follow-up treated with IFN in monotherapy were studied. We evaluated three kinds of response: sustained response (SR); non response (NR), and partial response (PR). Patients were evaluated biochemically and histologically by means of aminotransferase levels (pretreatment and serial) and hepatic biopsy (pretreatment and control). The data were obtained from clinical records. Aminotransferase levels at the end of treatment and six months later as well as histologic damage score showed the following results: 16 patients were cataloged as NR (61.5%), four as PR (15.3%), and six as SR (23.1%). Ten patients developed cirrhosis during follow-up, eight from NR group, one from PR group, and one from SR group. CHC patients treated with IFN alone obtained SR in 19% of cases; the patient who developed cirrhosis was not included. A study for longer time, prospective and protocolized to evaluate standardized IFN dose as well as evaluate combined treatment, is necessary.

A novel active DNA topoisomerase I in Leishmania donovani.

A common feature shared by type I DNA topoisomerases is the presence of a "serine, lysine, X, X, tyrosine" motif as conventional enzyme active site. Preliminary data have shown that Leishmania donovani DNA topoisomerase I gene (LdTOP1A) lacked this conserved motif, giving rise to different theories about the reconstitution of an active DNA topoisomerase I in this parasite. We, herein, describe the molecular cloning of a new DNA topoisomerase I gene from L. donovani (LdTOP1B) containing the highly conserved serine, lysine, X, X, tyrosine motif. DNA topoisomerase I activity was detected only when both genes (LdTOP1A and LdTOP1B) were co-expressed in a yeast expression system, suggesting the existence of a dimeric DNA topoisomerase I in Leishmania parasites.

Leishmania donovani methionine adenosyltransferase. Role of cysteine residues in the recombinant enzyme.

Methionine adenosyltransferase (MAT, EC 2.5.1.6)-mediated synthesis of S-adenosylmethionine (AdoMet) is a two-step process consisting of the formation of AdoMet and the subsequent cleavage of the tripolyphosphate (PPPi) molecule, a reaction induced, in turn, by AdoMet. The fact that the two activities, AdoMet synthesis and tripolyphosphate hydrolysis, can be measured separately is particularly useful when the site-directed mutagenesis approach is used to determine the functional role of the amino acid residues involved in each. The present report describes the cloning and subsequent functional refolding, using a bacterial expression system, of the MAT gene (GenBank accession number AF179714) from Leishmania donovani, the etiological agent of visceral leishmaniasis. The absolute need to include a sulfhydryl-protection reagent in the refolding buffer for this protein, in conjunction with the rapid inactivation of the functionally refolded protein by N-ethylmaleimide, suggests the presence of crucial cysteine residues in the primary structure of the MAT protein. The seven cysteines in L. donovani MAT were mutated to their isosterical amino acid, serine. The C22S, C44S, C92S and C305S mutants showed a drastic loss of AdoMet synthesis activity compared to the wild type, and the C33S and C47S mutants retained a mere 12% of wild-type MAT activity. C106S mutant activity and kinetics remained unchanged with respect to the wild-type. Cysteine substitutions also modified PPPi cleavage and AdoMet induction. The C22S, C44S and C305S mutants lacked in tripolyphosphatase activity altogether, whereas C33S, C47S and C92S retained low but detectable activity. The behavior of the C92S mutant was notable: its inability to synthesize AdoMet combined with its retention of tripolyphosphatase activity appear to be indicative of the specific involvement of the respective residue in the first step of the MAT reaction.