Construction and analysis of EST libraries of the trans-polyisoprene producing plant, Eucommia ulmoides Oliver.

Eucommia ulmoides Oliver is one of a few woody plants capable of producing abundant quantities of trans-polyisoprene rubber in their leaves, barks, and seed coats. One cDNA library each was constructed from its outer stem tissue and inner stem tissue. They comprised a total of 27,752 expressed sequence tags (ESTs) representing 10,520 unigenes made up of 4,302 contigs and 6,218 singletons. Homologues of genes coding for rubber particle membrane proteins that participate in the synthesis of high-molecular poly-isoprene in latex were isolated, as well as those encoding known major latex proteins (MLPs). MLPs extensively shared ESTs, indicating their abundant expression during trans-polyisoprene rubber biosynthesis. The six mevalonate pathway genes which are implicated in the synthesis of isopentenyl diphosphate (IPP), a starting material of poly-isoprene biosynthesis, were isolated, and their role in IPP biosynthesis was confirmed by functional complementation of suitable yeast mutants. Genes encoding five full-length trans-isoprenyl diphosphate synthases were also isolated, and two among those synthesized farnesyl diphosphate from IPP and dimethylallyl diphosphate, an assumed intermediate of rubber biosynthesis. This study should provide a valuable resource for further studies of rubber synthesis in E. ulmoides.

Externalization of saw-tooth architecture in small serrated polyps implies the presence of methylation of IGFBP7.

INTRODUCTION: Serrated polyps have been considered to be precursors of colorectal cancer with microsatellite instability. However, the biological and/or morphological changes which occur during the course of serrated polyp to cancer remain to be elucidated. METHODS: Twenty-eight colorectal serrated polyps including five mixed polyps (MP) from 20 patients were observed by chromoendoscopy with magnification, and subsequently resected endoscopically. The presence of mutations in two genes (K-ras and BRAF) and the methylation status of six genes (MLH1-A, MLH1-C, ESR1, P16, SOCS1, and IGFBP7) were examined. RESULTS: The 28 polyps included 32 histological serrated lesions (22 sessile serrated adenomas [SSA], six hyperplastic polyps [HP], and four traditional serrated adenoma [TSA]-like lesions). BRAF mutation was frequently observed in SSAs (19/22), while K-ras mutation was dominant in HPs (5/6). The externalization of saw-tooth architecture in serrated polyps was endoscopically observed more frequently in those with high levels of IGFBP7 methylation (P = 0.03). Moreover, the endoscopic finding was observed in five of six small serrated lesions (<10 mm) which contained both BRAF mutation and high levels of IGFBP7 methylation. TSA-like lesions in small MPs demonstrated the endoscopic finding with no or little MLH1 methylation, while the counterparts in the mixed polyps had high levels of MLH1 methylation with relatively low levels of IGFBP7 methylation. CONCLUSIONS: Our data suggests two distinct pathways may be involved in the early stages of the serrated pathway: one where MLH1 is primarily methylated, and a second where methylated IGFBP7 is associated with an externalization of saw-tooth architecture.

Human VIP-alpha: a long-acting, biocompatible and biodegradable peptide nanomedicine for essential hypertension.

We have previously shown that self-association of human vasoactive intestinal peptide with sterically stabilized liposomes (VIP-alpha) alters peptide conformation from random coil in aqueous solution to alpha-helix. This, in turn, protects the peptide from hydrolysis and amplifies and prolongs its bioactivity. The purpose of this study was to determine whether a single, intravenous injection of low-dose human VIP-alpha normalizes systemic arterial pressure in anesthetized spontaneously hypertensive hamsters for a prolonged period of time in a selective fashion. We found that intravenous injection of human VIP-alpha, VIP alone (each, 1.0 nmol) and empty liposomes had no significant effects on mean arterial pressure (MAP) in normotensive hamsters. By contrast, human VIP-alpha (0.01-1.0 nmol) evoked a significant concentration-dependent decrease in MAP to the normative range in spontaneously hypertensive hamsters that lasted throughout the observation period (6 h; p<0.05). VIP alone and empty liposomes had no significant effects on MAP in these animals. We conclude that a single, low-dose intravenous injection of human VIP-alpha normalizes systemic arterial pressure in spontaneously hypertensive hamsters for a prolonged period of time in a selective fashion. We suggest that human VIP-alpha should be further developed as a long-acting, biocompatible and biodegradable peptide nanomedicine for essential hypertension.

Triple therapy of interferon and ribavirin with zinc supplementation for patients with chronic hepatitis C: a randomized controlled clinical trial.

AIM: To study the therapeutic effect of interferon (IFN) and ribavirin with zinc supplement on patients with chronic hepatitis C viral (HCV) infection. METHODS: A total of 102 patients confirmed histologically to have chronic HCV infection with genotype 1b and more than 100 KIU/mL of HCV were randomly assigned to each arm of the study and each received 10 million units of pegylated interferon (IFN-alpha-2b) daily for 4 wk followed by the same dose every other day for 20 wk plus ribavirin (600 or 800 mg/d depending on body weight), with or without polaprezinc (150 mg/d) orally for 24 wk. The primary endpoint was sustained virological response (SVR) defined as negative HCV-RNA in the serum 6 mo after treatment. RESULTS: There were no differences in the clinical background between the two groups except for more females in the dual therapy group than in the other group (P<0.05). SVR was observed in 33.3% of the triple therapy group and 33.3% of the dual therapy group. The side effects were almost the same in both groups except for gastrointestinal symptoms, which were less in the triple therapy group (P=0.019). CONCLUSION: Considered together, triple therapy of zinc plus IFN and ribavirin for HCV infection patients with genotype 1b and high viral load is not better than dual therapy except for lower incidence of gastrointestinal side effects.

Are sterile water and brushes necessary for hand washing before surgery in Japan?

PURPOSE: To examine whether sterile water and brushes are necessary for hand washing before surgery. METHOD: Twenty-two operating room nurses were randomly divided into two groups as follows: 11 nurses who used 7.5% povidone iodine (PVI group) and another 11 nurses who used 4% chlorhexidine gluconate (CHG group) to wash their hands using the rubbing method. All the nurses were examined for bacterial contamination of their hands before and after surgical hand rubbing. We used tap water to wash the hands at the sink used for washing surgical instruments in the operating room and non sterilized plastic brushes. RESULTS: No bacteria were detected in the tap water. Before washing the hands, the number of bacteria detected was 5.0 x 10(3) cfu/H in the PVI group and 4.0 x 10(3) cfu/H in the CHG group, which were similar in both groups. After washing the hands, the median value of the bacteria decreased to 8.7 x 10(2) cfu/H in the PVI group and 0 cfu/H in the CHG group. CONCLUSIONS: Sterile water and brushes are not necessary for preoperative scrubbing up. When using tap water for surgical hand washing, 1) the hand-rubbing method should be used; 2) a quick-alcohol-based disinfectant scrub should be used; 3) the concentration of free chloride in the water should be maintained at over 0.1 PPM; 4) the bacterial contamination of the water should be checked; and 5) the faucet should be routinely cleaned and sterilized.

Abdominal incisional hernia repair using the Composix Kugel Patch: two case reports.

We describe two patients with abdominal incisional hernias, which occurred after appendectomy and replacement of an artificial blood vessel. Both were treated by operative hernial repair with the Composix Kugel Patch (C.R. Bard Inc.), a composite mesh that combines polypropylene mesh and expanded polytetrafluoroethylene (Gore-Tex). The mesh has various beneficial characteristics. It is a reinforcing material for the abdominal wall; even when in direct contact with the intestinal tract it is minimally adherent to the intestinal tract. The mesh expands readily and is easily fixed to the abdominal wall because it has a shape-memory ring. The long-term results of operative repair with this mesh have not yet been reported, but it is hoped that the aforementioned characteristics will yield favorable outcomes.

Identification of laticifer-specific genes and their promoter regions from a natural rubber producing plant Hevea brasiliensis.

Latex, the milky cytoplasm of highly differentiated cells called laticifers, from Hevea brasiliensis is a key source of commercial natural rubber production. One way to enhance natural rubber production would be to express genes involved in natural rubber biosynthesis by a laticifer-specific overexpression system. As a first step to identify promoters which could regulate the laticifer-specific expression, we identified random clones from a cDNA library of H. brasiliensis latex, resulting in 4325 expressed sequence tags (ESTs) assembled into 1308 unigenes (692 contigs and 617 singletons). Quantitative analyses of the transcription levels of high redundancy clones in the ESTs revealed genes highly and predominantly expressed in laticifers, such as Rubber Elongation Factor (REF), Small Rubber Particle Protein and putative protease inhibitor proteins. HRT1 and HRT2, cis-prenyltransferases involved in rubber biosynthesis, was also expressed predominantly in laticifers, although these transcript levels were 80-fold lower than that of REF. The 5'-upstream regions of these laticifer-specific genes were cloned and analyzed in silico, revealing seven common motifs consisting of eight bases. Furthermore, transcription factors specifically expressed in laticifers were also identified. The common motifs in the laticifer-specific genes and the laticifer-specific transcription factors are potentially involved in the regulation of gene expression in laticifers.