Effect of Lactobacillus plantarum enteral feeding on the gut permeability and septic complications in the patients with acute pancreatitis.

OBJECTIVE: To study the effect of the Lactobacillus plantarum (LP) enteral feeding on the gut permeability and sepsis in the patients with acute pancreatitis. SUBJECTS: Seventy-six subjects who stayed over 1 week in the hospital completed the study. Subjects were not treated with any lactobacillus supplement before the intervention. METHODS: Seventy-six patients with acute pancreatitis were randomly divided into two groups, parenteral nutrition (PN) group (n=38) and ecoimmunonutrition (EIN) group supplied by LP enteral feeding (n=36). The acute physiology and chronic health evaluation score, Balthazar CT score, CRP, fecal bacterial species and DNA fingerprint profiles as well as the potentially pathologenic organisms in nasogastric aspirate were determined on the day of admission and on the 8 day. The intestinal permeability was assessed by measurement of the ratio of lactulose/rhamnose on the day of admission and on days 5 and 8. The rate of organ failure, septic complications and death cases were evaluated at the 8 day. RESULTS: Following 7 days treatment, 38.9% patients in the EIN group were colonized with multiple organisms compared to 73.7% in the PN group (P<0.01), and 30.6% patients in the EIN grew potentially pathogenic organisms compared to 50% patients in PN group (P<0.05). The fecal bacterial DNA fingerprint profiles were less, the amount of lactobacteria and bifidobacteria decreased, and the amount of enterococci increased in PN group as compared with EIN group, P<0.05. By day 8, the lactulose/rhamnose ratio in EIN group were lower than that in PN group at days 5 and 8, P<0.05. The patients with LP got a better clinical outcomes as compared with the patients with PN. CONCLUSION: EIN enteral feeding can attenuate disease severity, improve the intestinal permeability and clinical outcomes.

Parathyroid hormone-related protein and calcium regulation in vitamin D-deficient sea bream (Sparus auratus).

Gilthead sea bream (Sparus auratus L.) were fed a vitamin D-deficient diet for 22 weeks. Growth rate, whole body mineral pools and calcium balance were determined. Plasma parathyroid hormone-related protein (PTHrP) and calcitriol levels were assessed. Expression of mRNA for pthrp and pth1r was quantified in gills and hypophysis. Fish on vitamin D-deficient diet (D- fish) showed reduced growth and lower calcium turnover (calcium influx, efflux and accumulation rates decreased) and unaltered plasma calcium levels. Plasma calcitriol levels became undetectable, PTHrP levels decreased in the D- fish. In controls, a significant increase in plasma PTHrP level over time was seen, i.e. it increased with body mass. Relationships were found between plasma PTHrP and the whole body pools of calcium, phosphorus and magnesium, indicative of a role for PTHrP in bone development. Expression of pthrp and pth1r mRNA was down-regulated in the hypophysis of D-fish, whereas in gill tissue, pthrp and pth1r mRNA were up-regulated. We conclude that lower pthrp mRNA expression and plasma values in D- fish reflect lower turnover of PTHrP under conditions of hampered growth; up-regulation of pthrp mRNA in gills indicate compensatory paracrine activity of PTHrP during calcitriol deficiency to guarantee well-regulated branchial calcium uptake. This is the first report to document a relation between PTHrP and calcitriol in fish.

Measurement of PTHrP, PTHR1, and CaSR expression levels in tissues of sea bream (Sparus aurata) using quantitative PCR.

A quantitative PCR (Q-PCR) method has been established to measure the mRNA expression levels of parathyroid hormone-related protein (PTHrP), parathyroid hormone receptor type 1 (PTHR1), and calcium-sensing receptor (CaSR) in sea bream (Sparus aurata), using the housekeeping gene, beta-actin, as endogenous control. TaqMan primers and probes were designed using the Primer Express program, according to the published/unpublished sequences of the three target genes and beta-actin of sea bream. Different tissues including gill, kidney, duodenum, hindgut, rectum, liver, heart, brain, pituitary, skin, muscle, and gonad were removed and immediately snap-frozen from three juvenile sea bream (100-150 g) cultured in sea water. The mRNAs were extracted and reverse-transcribed into cDNAs, which were subsequently examined by the ABI 5700 system using an optimized Q-PCR method. Triplicate measures of each sample indicated consistency of the technique. However, the mRNA expression levels for each transcript in these tissues were variable between fish and also relatively low. Nevertheless, this methodology can be used in the future studies of factors that may alter gene expression in these tissues.

[PCR amplification of anaerobic fungal 18S rDNA from landfill sites].

Nucleic acid based techniques are widely used to characterize microbial communities in environmental samples. In this study, seven landfill leachates were tested for the presence of 18S rRNA genes from anaerobic fungi from the family Chytridiomycetes. A nested Polymerase Chain Reaction (PCR) strategy was employed to amplify the 18S rRNA gene region of Chytridiomycetous fungi from landfill sites samples. All of the PCR products from landfill leachates amplified with the first primer pair NS1/NS8 and the second primer pair Chyt-719/Chyt-1553 were subjected to agarose gel electrophoresis to show seven samples containing fungi from the family Chytridiomycetes. One of the positive results was then selected for cloning and sequencing of the PCR product to confirm the identity of the amplified DNA. The molecular data presented here reveals for the first time that Chytridiomycetous fungi associated with the anaerobic rumen environment are also present in landfill sites.