Myenteric nitrergic neurons along the rat esophagus: evidence for regional and strain differences in age-related changes

Several studies have suggested an age-related reduction in the number of myenteric neurons in the lower gastrointestinal (GI) tract linked to changes in GI neuromuscular functions. The present study, combining protein gene product 9.5 immunostaining and NADPH-diaphorase histochemistry, aimed at quantifying the proportion of nitrergic neurons compared to the overall number of enteric neurons in the esophagus of young (3-4.5 months) and aged (18-20 months) Sprague-Dawley and Wistar rats. In both strains, the neuron numbers per ganglion in the cervical region were almost twice as high as in the other esophageal regions. Irrespective of age or strain, the esophagus harbored a very high proportion of intrinsic nitrergic neurons (greater than approximately 65%). Both strains showed with aging an overall neuronal loss of approximately 27%. While a significant increase (young: 64-71%; aged: 82-89%) was observed in all esophageal regions in the Wistar strain, the proportion of nitrergic neurons remained stable with aging in the Sprague-Dawley strain (range: 72-82%). In conclusion, the age-related reduction in the overall number of myenteric, nitrergic, and non-nitrergic neurons observed in the rat esophagus, appears to be highly region- and strain-dependent. Therefore, a protective mechanism against neuronal cell loss, selectively present in specific (nitrergic) enteric subpopulations, as suggested in earlier reports, cannot be put forward as a general phenomenon throughout the entire GI tract.     

Quantification of tagitinin C in Tithonia diversifolia by reversed-phase high-performance liquid chromatography

A simple, rapid and reliable reversed-phase high-performance liquid chromatographic method for the determination of tagitinin C, an anti-plasmodial sesquiterpene lactone isolated from the aerial parts of Tithonia diversifolia, has been developed. The assay has been used to quantify tagitinin C in various extracts of the aerial parts of T. diversifolia.     

Biodiversity of exopolysaccharides produced by Streptococcus thermophilus strains is reflected in their production and their molecular and functional characteristics

Twenty-six lactic acid bacterium strains isolated from European dairy products were identified as Streptococcus thermophilus and characterized by bacterial growth and exopolysaccharide (EPS)-producing capacity in milk and enriched milk medium. In addition, the acidification rates of the different strains were compared with their milk clotting behaviors. The majority of the strains grew better when yeast extract and peptone were added to the milk medium, although the presence of interfering glucomannans was shown, making this medium unsuitable for EPS screening. EPS production was found to be strain dependent, with the majority of the strains producing between 20 and 100 mg of polymer dry mass per liter of fermented milk medium. Furthermore, no straightforward relationship between the apparent viscosity and EPS production could be detected in fermented milk medium. An analysis of the molecular masses of the isolated EPS by gel permeation chromatography revealed a large variety, ranging from 10 to >2,000 kDa. A distinction could be made between high-molecular-mass EPS (>1,000 kDa) and low-molecular-mass EPS (<1,000 kDa). Based on the molecular size of the EPS, three groups of EPS-producing strains were distinguished. Monomer analysis of the EPS by high-performance anion-exchange chromatography with amperometric detection was demonstrated to be a fast and simple method. All of the EPS from the S. thermophilus strains tested were classified into six groups according to their monomer compositions. Apart from galactose and glucose, other monomers, such as (N-acetyl)galactosamine, (N-acetyl)glucosamine, and rhamnose, were also found as repeating unit constituents. Three strains were found to produce EPS containing (N-acetyl)glucosamine, which to our knowledge was never found before in an EPS from S. thermophilus. Furthermore, within each group, differences in monomer ratios were observed, indicating possible novel EPS structures. Finally, large differences between the consistencies of EPS solutions from five different strains were assigned to differences in their molecular masses and structures.     

Structure of gene coding for the fruit body-specific hydrophobin Fbh1 of the edible basidiomycete Pleurotus ostreatus

Two fruit body-specific hydrophobins (Fbh1 and POH1) have been identified in two different strains of the edible basidiomycete Pleurotus ostreatus. Comparison of their nucleotide and amino acid sequences yielded similarity values (59% and 66%, respectively) smaller than those found for alleles of the same hydrophobin gene but higher than those found for different hydrophobin genes in P. ostreatus var. florida (Pe?as et al 2002). In this paper, we have addressed the question of Fbh1 and POH1 allelism by studying the structure of the gene fbh1 and by a classical genetic analysis to compare it with that of POH1. The structure of both genes is similar, as revealed by the similarity of their promoters and leader peptide sequences and by the conserved position of their introns. Furthermore, the allelism analysis revealed that both genes segregated as alleles when present in the same hybrid. These results suggest an allelic condition for POH1 and fbh1 and stress the importance of the similarity of fbh1/POH1 promoter and leader sequences. Furthermore, we have identified various microsatellite-like regions in this gene that can be used for strain and species typing in the future.     

Reconstitution of lethally irradiated mice by cells isolated from adipose tissue

It is suggested that hematopoietic stem cells (HSC) could be found in several tissues of mesodermic origin. Among these, adipose tissue can expand throughout adult life and its expansion is not only due to mature adipocyte hypertrophy but also to the presence of precursor cells in stroma-vascular fraction (SVF). Here we report that transplantation of cells isolated from mice adipose tissue can efficiently rescue lethally irradiated mice and results in a reconstitution of major hematopoietic lineages. Donor cells can be detected in blood and in hematopoietic tissues of recipient mice. Adipose tissue contains a significant percentage of CD34, CD45 positive cells, and SVF cells were able to give rise to hematopoietic colonies in methylcellulose. We demonstrate the presence of hematopoietic progenitors in adipose tissue by phenotypic and functional characteristics. Thus adipose tissue could be considered as an important and convenient source of cells able to support hematopoiesis.     

Pharmacological sensitivity of ATP release triggered by photoliberation of inositol-1,4,5-trisphosphate and zero extracellular calcium in brain endothelial cells

Recently, ATP has gained much interest as an extracellular messenger involved in the communication of calcium signals between cells. The mechanism of ATP release is, however, still a matter of debate. In the present study we investigated the possible contribution of connexin hemichannels or ion channels in the release of ATP in GP8, a rat brain endothelial cell line. Release of ATP was triggered by photoactivation of InsP(3) or by reducing the extracellular calcium concentration. Both trigger protocols induced ATP release significantly above baseline. InsP(3)-triggered ATP release was completely blocked by alpha-glycyrrhetinic acid (alpha-GA), the connexin mimetic peptides gap 26 and 27, and the trivalent ions gadolinium and lanthanum. ATP release triggered by zero calcium was, in addition to these substances, also blocked by flufenamic acid (FFA), niflumic acid, and NPPB. Gap 27 selectively blocked zero calcium-triggered ATP release in connexin-43 transfected HeLa cells, while having no effect in wild-type and connexin-32 transfected cells. Of all the agents used, only alpha-GA, FFA and NPPB significantly reduced gap junctional coupling. In conclusion, InsP(3) and zero calcium-triggered ATP release show major similarities but also some differences in their sensitivity to the agents applied. It is suggested that both stimuli trigger ATP release through the same mechanism, which is connexin-dependent, permeable in both directions, potently blocked by connexin mimetic peptides, and consistent with the opening of connexin hemichannels.     

Differential cytoplasmic and whole cellular expression of histone H1 within the avian bursa of Fabricius

Bursal anti-steroidogenic peptide (BASP), purified from the chicken bursa of Fabricius (BF), has been previously demonstrated to be a potent and efficacious inhibitor of steroid hormone biosynthesis from chicken ovarian, and both mammalian and avian adrenal cells in vitro. Other studies have demonstrated that BASP can markedly reduce avian and mammalian mitogen-stimulated lymphocyte proliferation. Recent studies have indicated that BASP has a structural and functional relationship with histone H1. Immunohistochemical studies using a monoclonal antibody, which is known to recognize a common histone H1 epitope from several plant and animal species identified the protein within the cytoplasm and nucleus of distinct cells within both the cortex and medulla of all BF follicles. Additionally, epithelial cells within the BF expressed the protein strongly in the cytoplasm with reduced nuclear staining. In contrast, the same antibody did not recognize the protein in thymus of the same animals. The differential expression of histone H1 immunoreactivity within selected cells of the BF may support a previous proposed role of histone H1 in extranuclear and extracellular signaling in chickens and possibly other species.     

Onychomycosis by common non-dermatophyte moulds

Between 1990 and 2000, The Medical Mycology Laboratory at University of Antioquia (Medellín, Colombia), attended 4621 patients, with nail dystrophies. We used the direct test and culture of nail samples for diagnostic purposes. This report includes 310 patients, 7% (310/4621), with positive cultures for the same non-dermathophyte fungi, in five or more places of inoculation. From the 310 positive cultures, 284 (92%) were positive in direct test. No characteristic morphology for any of the genera could be detected. Until 1989 we had an average of 9 cases per year and between 1990 and 2000 we found 28 cases per year (an increase of 211%). The implicated genera were: Fusarium 50%, Nattrassia 31%, Aspergillus and Dendrophoma 7% each, Penicillium and Scopulariopsis contributed with 2% each, and Acremonium was found in less of 1% of cases. The general prevalence of onychomycosis due to non-dermatophytic fungi was about 12,4% between 1990-2000. This report describes the main epidemiological characteristics found, comparing them with results from other authors, and defines the basic profile of affected patients: adults (31-40 years old) who use occlusive shoes (trainers) and practice sport, which predispose to hyperhydrosis and consequently to onychomycosis. The course of the disease takes in average 30 months in both sexes. The percentage of onychomycosis found was 62% for women and 38% for men.     

Novel mechanism for high-altitude adaptation in hemoglobin of the Andean frog Telmatobius peruvianus

In contrast to birds and mammals, no information appears to be available on the molecular adaptations for O(2) transport in high-altitude ectothermic vertebrates. We investigated Hb of the aquatic Andean frog Telmatobius peruvianus from 3,800-m altitude as regards isoform differentiation, sensitivity to allosteric cofactors, and primary structures of the alpha- and beta-chains, and we carried out comparative O(2)-binding measurements on Hb of lowland Xenopus laevis. The three T. peruvianus isoHbs show similar functional properties. The high O(2) affinity of the major component results from an almost complete obliteration of chloride sensitivity, which correlates with two alpha-chain modifications: blockage of the NH(2)-terminal residues and replacement by nonpolar Ala of polar residues Ser and Thr found at position alpha131(H14) in human and X. leavis Hbs, respectively. The data indicate adaptive significance of alpha-chain chloride-binding sites in amphibians, in contrast to human Hb where chloride appears mainly to bind in the cavity between the beta-chains. The findings are discussed in relation to other strategies for high-altitude adaptations in amphibians.