The most biofilm forming bacteria in catheter, Esctherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus were isolated and identified from a patient's catheter occuring catheter-associated urinary tract infection (CA-UTI). We examined mRNA expression and its quantification of AIs synthetic genes encoding signal substance of quorum sensing from each bacterial species in order to elucidated quorum sensing mechanism. Both pure cultures for each bacterial strains and a mixed cultures with three were grown for 24 hr and 30 days. Initial densities to be able to detect mRNA expression oil single strains culture were shown at $2.4{\times}10^5$ CFU/ml, $5.4{\times}10^6$ CFU/ml of E. coli for ygaG and S. aureus for luxS, and at $6.9{\times}10^4$ CFU/ml of P. aeruginosa for rhlI and lasI. Also, in mixed culture of three, initial cell densities of mRNA expression were appear to at $7.3{\times}10^5$ CFU/ml, $1.6{\times}10^7$ CFU/ml of E. coli for ygaG and S. aureus for luxS, and at $2.1{\times}10^5$ CFU/ml of P. aeruginosa for rhlI and lasI. Each AIs synthetic gene was expressed in initial cell density and the mRNA expression of the genes were detected continously during 30 days. And then, the quantification of mRNA expression level of ygaG, rhlI, last, and luxS which were related AIs synthesis was done each time point by real-time RT-PCR. Interestingly, the mRNA levels of ygaG, rhlI, lasI, and luxS from the mixed culture was higher than those from each single strain culture. In the case of E. coli ygaG, the amount of transcript from the mixed culture was at least 30 times for that from single culture. In the case of P. aeruginosa rhlI and lasI, the amount of transcript from the mixed culture was at least 40 times and 250 times for that from single strain culture. In the case of S. aureus luxS, the amount of transcript from the mixed culture was at least 5 times for that from single strain culture. And specially, the mRNA expression of rhlI and lasI of P. aeruginosa showed the highest efficency among four AIs synthetic genes.
The aim of the present study was to elucidate the direct effects of melatonin on bladder activity and to determine the mechanisms responsible for the detrusor activity of melatonin in the isolated rat bladder. We evaluated the effects of melatonin on the contractions induced by phenylephrine (PE), acetylcholine (ACh), bethanechol (BCh), KCl, and electrical field stimulation (EFS) in 20 detrusor smooth muscle samples from Sprague-Dawley rats. To determine the mechanisms underlying the inhibitory responses to melatonin, melatonin-pretreated muscle strips were exposed to a calcium channel antagonist (verapamil), three potassium channel blockers [tetraethyl ammonium (TEA), 4-aminopyridine (4-AP), and glibenclamide], a direct voltage-dependent calcium channel opener (Bay K 8644), and a specific calcium/calmodulin-dependent kinase II (CaMKII) inhibitor (KN-93). Melatonin pretreatment ($10^{-8}{\sim}10^{-6}M$) decreased the contractile responses induced by PE ($10^{-9}{\sim}10^{-4}M$) and Ach ($10^{-9}{\sim}10^{-4}M$) in a dose-dependent manner. Melatonin ($10^{-7}M$) also blocked contraction induced by high KCl ($[KCl]_{ECF}$; 35 mM, 70 mM, 105 mM, and 140 mM) and EFS. Melatonin ($10^{-7}M$) potentiated the relaxation response of the strips by verapamil, but other potassium channel blockers did not change melatonin activity. Melatonin pretreatment significantly decreased contractile responses induced by Bay K 8644 ($10^{-11}{\sim}10^{-7}M$). KN-93 enhanced melatonin-induced relaxation. The present results suggest that melatonin can inhibit bladder smooth muscle contraction through a voltage-dependent, calcium-antagonistic mechanism and through the inhibition of the calmodulin/CaMKII system.
The purpose of this study was to examine the effects of dietary caffeine supplementation on bone mineral density and bone mineral content in ovariectomized rats. Twenty eight female Sprague-Dawley rats (body weight $210\;{\pm}\;5\;g$) were divided into two groups, ovariectomy (OVX) and Sham groups, which were each randomly divided into two subgroups that were fed control and control supplemented with caffeine diets (caffeine 0.03% diets). All rats were fed on experimental diet and deionized water ad libitum for 6 weeks. Bone mineral density (BMD) and bone mineral content (BMC) were measured using PIXImus (GE Lunar Co, Wisconsin) in spine and femur. Serum alkaline phosphatase activity (ALP) and osteocalcin and urinary DPD crosslinks value were measured as markers of bone formation and resorption. The results of this study indicate that body weight gain and food intake were higher in OVX groups than in Sham groups regardless of diets. There were no differences weight gain between the control and caffeine groups in both OVX and Sham groups. Within the OVX groups, serum Ca concentration was lower in rats fed caffeine than in rats fed the control diet. Serum ALP, osteocalcin, urinary Ca, and phosphate were not different in each group. Spine BMD, spine BMD/weight, and spine BMC/weight, femur BMD/weight and femur BMC/weight of ovariectomy groups were significantly lower than Sham groups. Within the OVX group, there were no differences in spine BMD and BMC and femur BMD and BMC. These results indicate that no significant differences in spine and femur BMD were found due to 0.03% caffeine intakes in diet in OVX rats for 6 weeks. No negative effect of caffeine in 0.03% diet on bone mineral density were found in the present study. Further investigation of the relation between caffeine and bone mineral density are warranted. (KoreanJNutr2008; 41(3): 2l6~223)
Bojungbangdocktang (BJBDT), a formula of eight Oriental herbs, is a modified herbal prescription of Bangdoktang and Bojungbangamtang. Recently, BJBDT was demonstrated to inhibit angiogenesis induced by vascular endothelial growth factor in human umbilical vein endothelial cells, enhance hematopoiesis and protect cisplatin-induced cytotoxicity in normal MCF-10A breast cells. Nevertheless, there is no safety study of BJBDT before clinical trial so far. Thus, in the current study, we investigated the toxicity about ethanol-extracted BJBDT. Male and female Spraque Dawley (SD) rats were given orally by BJBDT at 250, 500, and 1000 mg/kg for 4 weeks. Mortality, clinical signs and measured change of body weight, food consumption and water consumption were observed. In addition, we performed ophthalmologic, urinary, hematological, blood serum biochemical and histopathological examination. Any general toxicity was not found in BJBDT treated group. Also, there were no significant differences in the parameters such as body weight, food consumption and water consumption, a lot of urine and blood factor levels except HCT, MCHC, Ca, TG, Glucose and T-Bilirubin level compared with control group. Although HCT was elevated and TG was decreased in male rats, and MCHC, Glucose and T-Bilirubin were elevated and Ca and HCT were decreased in female rats, these were within normal ranges. Finally, we determined that maximum tolerated dose (MTD) was 1000 mg/kg and no observed adverse effect level (NOAEL) was 500 mg/kg. Taken together, these results demonstrated that BJBDT is very safe to SD rats.
This study explored the effect of calcium levels and/or ovariectomy on bone metabolism using female Sprague-Dawley weanling rats as a model . Rats received a low (0.1%) calcium diet for 8 weeks. The rats were then divided into three subgroups that were fed 0.1% ,0.5% and 1.5% calcium diets for 8 weeks after operation. The results of this experiment indicate that body weight gin was higher in ovariectomy groups than in sham groups regardless of calcium level and food intake. Serum Ca and P concentrations were of normal level regardless of calcium level and ovariectomy. Estrogen concentration was low in the ovariectomized group. Serum alkaline phophatase activity and urinary hydroxyproline have been used as markers of bone formation and resorption. These values were increased in ovariectomized groups. The weight, length and breaking force of femur were not significantly different between the groups. Ash, Ca, P and total lipid contents in femur and lumbar were decreased in the groups fed low calcium . Mg content was decreased in the ovariectomy and total protein content was not affected by calcium level and ovariectomy. The effect groups of ovarectomy on calcium contents of bone was more prominent in lumbar than in femur. In conclusion, though low calcium intakes during growth period may retard the attainment of peak bone mass, calcium supplementation after this period increased bone growth and mineral contents, but not significant effect in three calcium levels. Furthermore, calcium intake was shown to have a greater influence on the mineral contents of femur than of lumbar, and removal of endogenous estrogen production by ovariectomy was shown to be more deleterious on the ash and calcium contents of the lumbar than of femur.
This study was performed to evaluate the effect of red-yeast-rice on bone metabolism in overiectomized (OVX) rats. Forty female Sprague-Dawley rats (body weight 210 $\pm$ 5 g, 9 weeks old age) were divided into two groups. One group were OVX, and the other group received sham operation (SHAM), and received either control diet (20% casein) or a red-yeast-rice power supplemented diet (0.1%) for 9 weeks. And then each rat group was further divided into control diet (casein 20%) and red-yeast-rice powder supplemented (0.1%) diet group. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks. Bone mineral density (BMD) and bone mineral content (BMC) were measured using PIXImus (GE Lunar Co, Wisconsin, USA) in spine and femur on 5, 9 weeks after feeding. The serum and urine concentrations of Ca and P were determined. Bone formation was measured by serum osteocalcin and alkaline phosphatase (ALP) concentrations. And bone resorption rate was measured by deoxypyridinoline (DPD) crosslinks immunoassay and corrected for creatinine. Serum osteocalcin, growth hormone, IGF-l and calcitonin were analyzed using radioimmunoassay kits. Urinary Ca and P excretion were not significantly different among the groups. Within the OVX group, the red-yeast-rice group had a lower crosslinks value than the casein group. Therefore the red-yeast-rice supplemented groups had a lower bone resorption ratio than the casein group in the ovariectomized rats. And, the red-yeast-rice group had significantly higher IGF-l hormone than casein group in ovariectomized rats. The red-yeast-rice group had higher spine bone mineral content than those of control group within the OVX groups. This study was an important first step in establishing that the observed beneficial effects of red-yeast-rice on bone, and this study also established the need for a study on the long-term effect of this supplement in a human.
Journal of Physiology & Pathology in Korean Medicine
/
v.23
no.4
/
pp.799-804
/
2009
Kamikaekyuk-tang(KMKKT), a formula of ten Oriental herbs, was orientally designed to promote vital energy, to remove blood stasis, and to decrease inflammation for treating cancers. KMKKT and its component had potent antiandrogen and androgen receptor activities in prostate cancer and also inhibited angiogenesis induced by basic fibroblast growth factor (bFGF) in human umbilical vein endothelial cells and suppressed the tumor growth in LLC-bearing mice, and liver metastasis of colon 26-L5 cancer cells, suggesting a potent cancer preventive agent. Nevertheless, there is no safety study of KMKKT before clinical trial so far. Thus, in the current study, we investigated the toxicity about ethanol-extracted KMKKT. Male and female Spraque Dawley (SD) rats were given orally by KMKKT at 250, 500, and 1000 mg/kg for 4 weeks. Mortality, clinical signs and measured change of body weight, food consumption and water consumption were observed. In addition, we performed ophthalmologic, urinary, hematological, blood serum biochemical and histopathological examination. Any general toxicity was not found in KMKKT treated group. Also, there were no significant differences in the parameters such as body weight, food consumption and water consumption, a lot of urine and blood factor levels except WBC, MCHC and Ca level compared with control group. Although WBC and MCHC were elevated in female rats and Ca level was decreased in male rats, these were within normal ranges. Finally, we determined that maximum tolerated dose (MTD) was 1000 mg/kg and no observed adverse effect level (NOAEL) was 500 mg/kg. Taken together, these results demonstrated that KMKKT is very safe to SD rats.
Abouheif, M.A.;Al-Saiady, M.;Kraidees, M.;Eldin, A. Tag;Metwally, H.
Asian-Australasian Journal of Animal Sciences
/
v.13
no.7
/
pp.967-973
/
2000
A metabolism trial was conducted with 28 Najdi rams allocated into seven dietary groups to evaluate the effect of dietary inclusion of Salicornia bigelovii Torr biomass on nutrient digestibility, rumina I fluid metabolites and nitrogen and mineral balances. Either the stems (ST) or spikes (SP) of this seawater-irrigated halophyte were incorporated into complete diets at rates of 0, 10, 20 and 30% levels, replacing equal amounts of rhodesgrass hay in a ground mixed control diet. Digestibility of DM, OM, EE, NFE and fecal and urinary nitrogen were not affected by increased level of ST in the diet. As level of ST increased from 0 to 20% in the diets, CP digestibility and nitrogen retention approached their maximum (p<0.01), whereas CF digestibility reached its minimum (p<0.01). On the other hand, except for EE, digestion of all nutrients and nitrogen retention were linearly depressed (p<0.01) as SP increased in the diets from 10 to 30% level. Concentration of ammonia-N, total VFA and pH values in the rumen fluid were lower (p<0.01) with the ST- or SP-fed diets than with the control diet. Increasing level of ST or SP in the diet was associated with an increase (p<0.01) in the proportion of acetate and a decline (p<0.01) in molar percentage of propionate in the ruminal fluid. Sodium absorption increased (p<0.01) with increased ST and SP in the diets up to the 10 and 20% level, respectively, followed by constant absorption values up to the 30% level. When the level of ST in the diet gradually increased to 30%, a concomitant increased (p<0:01) in Ca and P absorption were obvious; whereas, increased level of SP in the diets from 0 to 30% resulted in noticeable (p<0.01) depression in Ca and P apparent absorption.
Purpose : The purpose of this study was to investigate whether hypercalciuria patients with hematuria show different renal indices compared to non-hypercalciuria patients with hematuria. Methods : We retrospectively reviewed the medical records of patients with gross or microscopic hematuria whose blood chemistry and 24 hour urine chemistry were examined. After excluding the patients with more than $4 mg/m^2/day$ proteinuria or the patients with urinary calcium excretion between 3 and 4 mg/kg/day, we divided the patients into two groups: a hypercalciuria group whose calcium excretion was more than 4 mg/kg/day(n=30) and a non hypercalciuria group whose calcium excretion was less than 3 mg/kg/day(n=41). The urinary excretion, clearance, and fractional excretion(FE) of Na, K, Cl, Ca, P, urea, and creatinine were calculated and compared between the two groups. Results : The hypercalciuria group had more calcium excretion($6.1{\pm}2.9$ vs $1.5{\pm}0.9 mg/kg/day$), more urea excretion($341{\pm}102$ vs $233{\pm}123 mg/kg/day$), greater glomerular filtration rate(GFR) ($93.7{\pm}31.1$ vs $79.5{\pm}32.0 mL/min$) but lower FENa($1.0{\pm}0.4%$ vs $1.3{\pm}0.6%$) than the nonhyper-calciuria group, although the urinary sodium excretion was similar between the two groups. Conclusion : The greater urea excretion and GFR in hypercalciuric patients suggest that they might be on a higher protein diet than the non-hypercalciuria group. The increased glomerular filtration of sodium and calcium induced by the higher GFR in hypercalciuria would have increased their delivery to the distal tubule, where sodium is effectively reabsorbed but calcium is not, which is suggested by the lower FENa but higher FECa in hyercalciuria. It is recommended that the diet of hematuria patients be reviewed in detail at initial presentation and during treatment.
This study was desiged to investigate the effects of calcium supplementation on the metabolism of sodium and potassium and blood pressure in seven healthy college women, aged from 19 to 21 years old. For this purpose, metabolic studies were conducted for two weeks. During the first week, the subjects ate experimental diet of which nutrients composition was similar to their usual intake. And during the consecutive second week, they ate the same experimental diet supplemented with 500mg or calcium daily. The results were summarized as follows : 1) Urinary excretion of sodium was significantly increased(p<0.05), but fecal excretion and retention of it was not affected by supplementary intake of calcium. 2) Potassium balance was not changed after additional intake of calcium. 3) Serum sodium and potassium level decreased significantly(p<0.05), but aldosterone and renin levels in serum were not changed by additional intake of calcium. 4) Systolic blood pressure(SBP) was not affected, but diastolic blood pressure (DBP) was significantly decreased (p<0.05) by supplementation of calcium. The above results showed that daily supplementary intake of calcium can be effective to decrease diastolic blood pressure through inducing the change of sodium metabolism in young women eating usual Korean diets.
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