Leticia Santos

Identification of yeast and bacteria involved in the mezcal fermentation of Agave salmiana

Aims:  To identify the yeast and bacteria present in the mezcal fermentation from Agave salmiana.

Expansion of human hematopoietic stem cells for transplantation: trends and perspectives

Umbilical cord blood transplantation is clinically limited by its low progenitor cell content. Ex vivo expansion has become an alternative to increase the cell dose available for transplants. Expansion has been evaluated in several ways such as static cultures combining growth factors or mimicking the natural microenvironment using co-culture systems. However, static cultures have a small volume capacity and therefore large-scale expansion has been addressed using bioreactors. These and other biotechnological approaches for the expansion of hematopoietic progenitors and their utility to study several aspects of hematopoietic stem cell biology are discussed here.

Isolation and phylogenetic classification of culturable psychrophilic prokaryotes from the Collins glacier in the Antarctica

Culturable psychrophilic prokaryotes were obtained of samples of glacier sediment, seaside mud, glacier melted ice, and Deschampsia antarctica rhizosphere from Collins glacier, Antarctica. The taxonomic classification was done by a culture-dependent molecular approach involving the Amplified Ribosomal DNA Restriction Analysis. Two hundred sixty colonies were successfully isolated and sub-cultivated under laboratory conditions. The analysis showed a bacterial profile dominated by Beta-proteobacteria (35.2%) followed by Gamma-proteobacteria (18.5%), Alpha-proteobacteria (16.6%), Gram-positive with high GC content (13%), Cytophaga–Flavobacterium–Bacteroides (13%) and Gram-positive with low GC content (3.7%). Eleven of the isolates have been reported previously and the others microorganisms remain uncharacterized. The isolated microorganisms here could be a potential source for biotechnological products, such as cold-active enzymes and secondary metabolites.

Identification of calcium stress induced genes in amaranth leaves through suppression subtractive hybridization

Calcium (Ca(2+)) is a critical ion for the growth and development of plants and plays an important role in signal transduction pathways in response to biotic and abiotic stresses. We investigated the Ca(2+) stress responsive-genes in amaranth leaves by using the suppression subtractive hybridization technique. Screening of the libraries generated 420 up-regulated transcripts and 199 down-regulated transcripts. The differentially expressed transcripts were associated with general stress response, transcription factors, gene regulation, signal transduction, and some other with unknown function. Selected genes were used to study their differential regulation by sqRT-PCR. Among the up-regulated transcripts, a fragment containing the motif of C3HC4-type RING-Zinc family was further characterized. The ORF of amaranth zinc finger protein (AhZnf) has a closer relationship with its ortholog from Ricinus communis while is distantly related to the Arabidopsis thaliana C3HC4-type ortholog. We have identified a novel putative zinc finger protein along with other novel proteins such as the wall associated kinase, phosphoinositide binding protein, and rhomboid protease involved in response to Ca(2+) stress in amaranth leaves.

Identification of differential expressed transcripts in cervical cancer of Mexican patients

The aim of this study was to identify the gene expression profile in biopsies of patients with cervical intraepithelial neoplasia (CIN) 1, CIN 2, CIN 3, and microinvasive cancer by suppression subtractive hybridization and Southern blotting. After analyzing 1,800 cDNA clones, we found 198 upregulated genes, 166 downregulated, and no significant change of gene expression in 86 clones (p = 0.005). These results were validated by Northern blot analysis (p = 0.0001) in the identification of 28 overexpressed and 7 downregulated transcripts. We observed a set of genes related to the Notch signaling pathway that may be involved in the transformation of cervical cells and in the development to malignancy. The differentially expressed genes may provide useful information about the molecular mechanisms involved in human cervical carcinoma and as diagnostic markers.

Vía de señalización Notch y nuevas estrategias para el tratamiento de cáncer

The Notch signaling pathway plays a crucial role at different stages of cell development, such as proliferation, growth, differentiation, and apoptosis. Recent studies demonstrate that depending on the expression level and cellular context, the Notch receptors play a role in apoptosis resistance in malignant cells. These findings suggest that Notch signaling components may be a potential target in the development of new cancer therapies. This review describes the function of the Notch pathway and new strategies in the modulation of its signal.

Characterization of ATPase activity of the AAA ARC from Bifidobacterium longum subsp. infantis.

Bifidobacteria are considered to be probiotics that exist in the large intestine and are helpful to maintain human health. Oral administration of bifidobacteria may be effective in improving the intestinal flora and environment, stimulating the immune response and possibly preventing cancer. However, for consistent and positive results, further well-controlled studies are urgently needed to describe the basic mechanisms of this microorganism. Analysis of the proteasome-lacking Bifidobacterium longum genome reveals that it possesses a gene, IPR003593 AAA ATPase core, which codes a 56 kDa protein containing one AAA ATPase domain. Phylogenetic classification made by CLANS, positioned this sequence into the ARC divergent branch of the AAA ATPase family of proteins. N-terminal analysis of the sequence indicates this protein is closely related to other ATPases such as the Rhodococcus erythropolis ARC, Archaeoglobus fulgidus PAN, Mycobacterium tuberculosis Mpa and the human proteasomal Rpt1 subunit. This gene was cloned, the full-length recombinant protein was overexpressed in Escherichia coli, purified as a high-molecular size complex and named Bl-ARC. Enzymatic characterization showed that Bl-ARC ATPase is active, Mg(+2)-dependent and sensitive to N-ethylmaleimide. Gene organization positions bl-arc in a region flanked by a cluster of genes that includes pup, dop and pafA genes. These findings point to a possible function as a chaperone in the degradation pathway via pupylation.

Protein Expression Analysis in Uterine Cervical Cancer for Potential Targets in Treatment

Specific markers in lesions of the human uterine cervix cancer (UCC) are still needed for prognostic, diagnostic and/or therapeutic purposes. In this study we evaluated key molecules at protein level between normal epithelium, cervical intraepithelial neoplasia (CIN1–3) and invasive cancer of a group of molecules previously reported at mRNA level. For that purpose, human formalin-fixed paraffin embedded tissue microarrays (TMAs) were constructed containing 205 Mexican tissue core specimens. Immunohistochemistry and quantitative analysis of histological staining was performed against twenty-two distinct proteins for each core and the processing platform ImageJ. In the progression of the disease we found key statistical differences for the proteins SEL1, Notch3 and SOCS3. High expressions of SEL1L, Notch3 and SOCS3 have potential value to increase the prognostic of UCC in combination with markers such as p16INK4a. This study identified key drivers in cervical carcinogenesis that should be evaluated for the development of UCC therapies.

Identification, heterologous expression and detection of enzymatic activity of an asparaginase from the archaeon Thermoplasma acidophilum

Abstract Asparaginases (ASNases) participate in the metabolism of all living organisms in the hydrolysis of free asparagines. Bacterial ASNases are used in cancer chemotherapy as they efficiently deplete amino acids. However, allergic reactions and silent inactivation represent critical limitations to their extended use. The rationale of this study was to identify, express, and characterize a plant-type L-ASNase from the archaeon Thermoplasma acidophilum as an enzyme with potentially improved characteristics. The Ta0338 orf was cloned into the pET28a(+) expression vector and overexpressed as a soluble protein with a molecular weight of 32 kDa. The quantity of recombinant L-ASNase produced in Escherichia coli was estimated as 9.68 mg/l. The purified protein showed evident autocatalytic processing of the zymogen at 4° and 37°C at physiological pH of 7.2 and clearly generated the expected alpha and beta subunits of 18 and 13 kDa, respectively. We propose that Ta-ASNase represents a potential biotechnological product for therapeutic purposes.