Enhanced Production of β-D-glycosidase and α-L-arabinofuranosidase in Recombinant <Emphasis Type="Italic">Escherichia coli</Emphasis> in Fed-batch Culture for the Biotransformation of Ginseng Leaf Extract to Ginsenoside Compound K

Ginsenoside compound K is an essential ingredient in nutritional supplements, cosmetics, and traditional medicines. However, cultivation for the production of enzymes involved in ginsenoside biotransformation has not been attempted in a fermenter. The host strain Escherichia coli ER2566 and the constitutive pHCE vector were selected for the efficient production of β-D-glycosidase, and expression medium composition to produce Sulfolobus solfataricus β-glycosidase expressed in E. coli was optimized in flask and batch cultures. The total activity of β-Dglycosidase in fed-batch culture using a fermenter increased 14-fold before optimization. S. solfataricus β-D-glycosidase and Thermotoga petrophila α-L-arabinofuranosidase were produced in a fed-batch culture. These two enzymes completely converted protopanaxadiol-type ginsenosides in ginseng leaf extract obtained from discarded ginseng leaves as a renewable substrate to compound K. The effective bioprocess for compound K production developed here will contribute to the industrial biological production of compound K.     

Serum Concentrations of Trace Elements Zinc,Copper, Selenium,and Manganese in Critically Ill Patients

Biological Trace Element Research - We measured serum concentrations of trace elements and evaluated their clinical significance in relation to treatment outcomes of critically ill patients. A...     

Use of helper-free retroviral vector to direct a high expression of porcine growth hormone in mouse fibroblast cells.

A retroviral vector has been employed to express the cDNA coding for porcine growth hormone (pGH) in the mouse fibroblast cell NIH 3T3 in large quantity. In this study, a single gene vector which contained no selectable marker was used. We have coinfected NIH 3T3 cells with pGH retrovirus and Neo(r) retrovirus to obtain a stable, high-expression clone. Using a superinfection strategy, we further increased the copy number of proviral DNA in the host chromosome, thus increasing the pGH secretion from 22 to 55 micrograms/10(6) cells/24 h. The recombinant pGH produced from mouse fibroblast cells was heterogeneous at the N-terminus, which mimicked the situation with bovine growth hormone either from natural sources or from recombinant products derived from mouse fibroblasts. This technology is useful for many biologically important genes to be stably transduced by retroviral vector into mammalian cells and highly expressed.     

Ginsenoside Rc and Rg1 Differentially Modulate NMDA Receptor Subunit mRNA Levels after Intracerebroventricular Infusion in Rats

We investigated the influence of centrally administered ginsenoside on the regulation of mRNA levels of the family of NMDA receptor subtypes (NR1, NR2A, NR2B, NR2C) by in situ hybridization histochemistry in the rat brain. The ginsenosides Rc and Rg1, the major components of ginseng saponin, differentially modulate NMDA receptor subunit mRNA levels in rat brain following prolonged i.c.v.-infusion. Ginsenosides Rc or Rg1 (10 g/10 l/hr for 7 days) was infused through preimplanted cannulae connected to osmotic mini-pumps. The level of NR1 mRNA is significantly increased in temporal cortex, caudate putamen, hippocampus, and granule layer of cerebellum in Rg1-infused rats as compared to control group. The level of NR2A mRNA is elevated in the frontal cortex. In contrast, it was decreased in CA1 area of hippocampus in Rg1-infused rats. However, there was no significant change of NR1 and NR2A mRNA levels in Rc-infused rats. The level of NR2B mRNA is elevated in cortex, caudate putamen, and thalamus in both Rc- and Rg-infused rats. In contrast, NR2B level is decreased in CA3 in Rg1-infused rats. The level of NR2C mRNA is increased in the granule layer of cerebellum in only Rg1 but not Rc infused rats. These results show that structure difference of ginsenoside may diversely affect the modulation of expression of NMDA receptor subunit mRNA after infusion into cerebroventricle in rats.     

Characteristics of nutrient intakes,basal physical fitness and serum markers in elderly women with osteopenia

Purpose

This study investigated the correlations among BMD, nutrient intake, basal physical fitness and serum markers in elderly women with osteopenia.

Methods

Bone Mineral Density (BMD) was measured in 54 elderly women (60–77yrs) by using dual energy X-ray absorptiometry (DEXA). The subjects were divided into an osteopenia group (OG; n = 29) and a normal group (NG; n = 25). A physical fitness test was conducted to determine muscle strength, muscle endurance, flexibility, agility, and balance. The dietary nutrient intake for one day was analyzed. The serum Ca, Pi and Mg concentration was also analyzed.

Results

There was significant correlation between All subject’s weight and their total-BMD as well as the T-score (p < .01). There was also significant correlation between the fat free mass and Total-BMD (p < .01). There was significant positive correlation between energy intake and dietary carbohydrate with Total-BMD and T-score in the OG (p < .05). The Serum Pi of the OG was significantly lower than that of the NG (p < .05), and there was significant correlation with the BMD in most regions as well as with the Total-BMD and T-score (p < .05, p < .01).

Conclusion

These results suggested that the fat free mass in elderly women plays an important role in maintaining their BMD and that sufficient caloric intake would be likely to improve BMD in elderly women with osteopenia. In addition, the results suggested the possibility of a decrease in bone mass by hypophosphatemia in elderly women.     

Innate immunity markers in culprit plaques of acute myocardial infarction or stable angina

We analyzed the innate immunity markers, C-reactive protein (CRP), serum amyloid P component (SAP) and pentraxin-3 (PTX-3), and metalloproteinase-9 (MMP-9) in coronary atherectomy tissues obtained from patients with acute ST elevation myocardial infarction (STEMI) (n=27) or stable angina (n=15). The relative areas immunopositive for CRP and SAP were similar in the two groups. In contrast, the proportion of areas immunopositive for PTX-3 and MMP-9 was higher in the STEMI group, compared to the stable angina group. PTX-3 or MMP-9-stained areas largely overlapped with those positive for CD68. We concluded that PTX-3 plays a role in the pathogenesis of STEMI.