Antiproteinuric Effects of Green Tea Extract on Tacrolimus-Induced Nephrotoxicity in Mice.

INTRODUCTION: It has been reported that proteinuria is an early predictive marker in detection of tacrolimus (TAC) nephrotoxicity. The aim of this study was to investigate the antiproteinuric effects of green tea extract (GTE) on TAC-induced acute nephrotoxicity in mice. METHODS: The mice (n = 20) were divided into 4 groups (n = 5 per group); control group mice were intraperitoneally (IP) injected with 0.9% saline, TAC group mice were IP injected with TAC 1 mg/kg, and inducible nitric oxide synthase (iNOS) inhibitor group mice were given in addition NG-nitro-L-arginine-methyl ester 12 mmol/L by subcutaneous injection. TAC-GTE group mice were given TAC by IP injection and GTE 100 mg/kg by subcutaneous injection. RESULTS: The 24-hour urine protein amounts were significantly increased in TAC group mice (36.1 ± 9.9 mg/d) compared with control group mice (13.3 ± 5.4 mg/d) and significantly decreased in TAC-GTE group mice (19.1 ± 6.9 mg/d, P < .01) compared with TAC group mice. The nitric oxide (NO) production by TAC was significantly suppressed by GTE and iNOS inhibitor injection. Renal tissue malondialdehyde (MDA) level was significantly increased in the TAC group compared with the control group and was significantly decreased in the TAC-GTE group compared with that of the TAC group. The antioxidant enzyme activities of superoxide dismutase and catalase were significantly suppressed in the TAC group compared with the control group and were restored in the GTE injection group. CONCLUSIONS: GTE treatment has beneficial antiproteinuric effects on TAC-induced acute renal injury in mice.

Rubi Fructus (Rubus coreanus) activates the expression of thermogenic genes in vivo and in vitro.

OBJECTIVE: To investigate the anti-obesity effect of Rubi Fructus (RF) extract using brown adipose tissue (BAT) and primary brown preadipocytes in vivo and in vitro. METHODS: Male C57BL/6 J mice (n=5 per group) were fed a high-fat diet (HFD) for 10 weeks with or without RF. Brown preadipocytes from the interscapular BAT of mice (age, post-natal days 1-3) were cultured with differentiation media (DM) including isobutylmethylxanthine, dexamethasone, T3, indomethacin and insulin with or without RF. RESULTS: In HFD-induced obese C57BL/6 J mice, long-term RF treatment significantly reduced weight gain as well as the weights of the white adipose tissue, liver and spleen. Serum levels of total cholesterol and low-density lipoprotein cholesterol were also reduced in the HFD group which received RF treatment. Furthermore, RF induced thermogenic-, adipogenic- and mitochondria-related gene expressions in BAT. In primary brown adipocytes, RF effectively stimulated the expressions of thermogenic- and mitochondria-related genes. In addition, to examine whether LIPIN1, a regulator of adipocyte differentiation, is regulated by RF, Lipin1 small interfering RNA (siRNA) and RF were pretreated in primary brown adipocytes. Pretreatment with Lipin1 siRNA and RF downregulated the DM-induced expression levels of thermogenic- and mitochondria-related genes. Moreover, RF markedly upregulated AMP-activated protein kinase. Our study shows that RF is capable of stimulating the differentiation of brown adipocytes through the modulation of thermogenic genes. CONCLUSIONS: This study demonstrates that RF prevents the development of obesity in mice fed with a HFD and that it is also capable of stimulating the differentiation of brown adipocytes through the modulation of thermogenic genes, which suggests that RF has potential as a therapeutic application for the treatment or prevention of obesity.

The antiproteinuric effects of green tea extract on acute cyclosporine-induced nephrotoxicity in rats.

BACKGROUND: It has been reported that the proteinuria is an early useful marker to detect cyclosporine (CsA) nephrotoxicity. The aim of this study was to investigate the antiproteinuric effects of green tea extract (GTE) on CsA-induced acute renal injury in rats. METHODS: The rats (n = 28) were divided into four groups (n = 7/group); controls intraperitoneally (IP) injected with 0.9% saline; CsA group IP injected CsA (50 mg/kg); inducible nitric oxide synthase (iNOS) inhibitor group administered in addition NG-nitro-L-arginine-methyl ester (12 mmol/L) subcutaneously and CsA-GTE group of CsA IP plus GTE (100 mg/kg) subcutaneously. RESULTS: The 24-hour urine proteins were significantly increased among the CsA (22.6 ± 3.1 mg/d) compared with the control (7.1 ± 1.5 mg/d) and significantly decreased in the CsA-GTE group (8.2 ± 1.8 mg/d, P < .01). Nitric oxide production induces by CsA treatment was significantly suppressed by GTE and iNOS inhibitor. Renal tissue malondialdehyde level was significantly increased in the CsA compared with controls and significantly decreased in the CsA-GTE group. The antioxidant enzyme activities of superoxide dysmutase and catalase, which were significantly suppressed in the CsA compared with the control group, were restored in the CsA-GTE cohort. CONCLUSION: GTE treatment of rats showed meaningful antiproteinuric effects through antioxidative activity in kidneys from CsA-induced acute renal injury.

Cottonseed with a high (+)- to (-)-gossypol enantiomer ratio favorable to broiler production.

This study was designed to evaluate the relative toxicity of (+)- and (-)-gossypol enantiomers in 0-3-week-old broilers. Treatments consisted of broiler starter diets formulated with either a glandless, which did not contain gossypol, a commercial glanded [62. 2% (+)-gossypol], or a glanded moco [83.2% (+)-gossypol] crushed cottonseed (CCS) (six replicates/treatment) plus a soybean meal negative control. Glandless cottonseed was mixed with the moco cottonseed (2.4% free gossypol) so that both the commercial glanded and moco glanded cottonseeds contained equivalent concentrations of free gossypol (2.0%). The cottonseed treatments were added at 5 and 10% of the diet. Body weights and feed conversions were determined weekly. Body weights and feed-to-gain ratios of broilers fed 5 and 10% glandless CCS and 5% moco CCS were not significantly different. Broilers receiving 10% commercial glanded CCS weighed significantly less than those subjected to all other treatments. Feed-to-gain ratios were significantly higher for broilers receiving 10% commercial glanded and 10% moco CCS as compared to 5% moco and glandless CCS, 10% glandless CCS, and control. Relative liver weights of birds receiving 10% moco CCS were significantly less than those of birds receiving 10% commercial CCS. The data clearly showed that broilers fed moco CCS containing a relatively high (+)- to (-)-gossypol enantiomer ratio performed better than broilers receiving commercial CCS with a lower (+)- to (-)-gossypol enantiomer ratio.

Effect of preoperative biofeedback/pelvic floor training on continence in men undergoing radical prostatectomy.

OBJECTIVES: To determine whether preoperative biofeedback training improves urinary continence overall or the rate of return of continence in men undergoing radical prostatectomy. METHODS: One hundred men scheduled to undergo radical prostatectomy were randomized to receive graded pelvic muscle exercise training with biofeedback 2 to 4 weeks before surgery or to a control group performing pelvic muscle exercises without biofeedback. The biofeedback group was instructed to continue exercises four times per day until surgery and to resume exercises when the urethral catheter was removed following surgery. The control group received written and brief verbal instructions in pelvic muscle exercises before surgery and again after catheter removal. Urinary continence was assessed by personal or phone interviews. RESULTS: Six months following surgery, the continence rates, as defined by the use of one pad or less per day, were 94% (44 of 47) and 96% (48 of 50) in the biofeedback and control groups, respectively (P = 0.596). Also, the rate of return as determined at time points 1, 2, 3, and 4 months after surgery was not significantly different between the two groups. CONCLUSIONS: Preoperative biofeedback training did not improve the outcome of pelvic muscle exercises on overall continence or the rate of return of urinary control in men undergoing radical prostatectomy.

Role of transurethral biopsy sampling of the prostate to diagnose prostate cancer in men undergoing surgical intervention for benign prostatic hyperplasia.

PURPOSE: Newer minimally invasive surgical procedures are being used to treat men with significant benign prostatic hyperplasia (BPH). These modalities do not allow retrieval of prostate tissue for histologic review. The goal of our study was to assess the value of transurethral biopsies in detecting prostate cancer in men who would undergo surgical intervention for BPH. MATERIALS AND METHODS: Between September 1997 and January 1999, 422 men undergoing transurethral resection of the prostate (TURP) had transurethral biopsies obtained before completing the TURP. Pathology reports as well as prostate-specific antigen (PSA) results were reviewed and analyzed to determine when cancer was present. RESULTS: Pathological examination revealed that cancer was found in 53 men (12.5%). The transurethral biopsies detected cancer in 32 of 53 (60.4%). No cancers were found in the transurethral biopsy specimen only. Of the 21 cancers missed by transurethral biopsy, 7 were stage T1b. PSA level >10 ng/mL increased the likelihood of finding cancer. CONCLUSIONS: Transurethral biopsy sampling is unreliable for detecting prostate cancer in men with clinically significant BPH. Significant cancers are missed if transurethral biopsies are used to determine the presence of carcinoma before minimally invasive surgical therapy for BPH.

St. John's wort for depression: a meta-analysis of well-defined clinical trials.

Studies concluding that St. John's wort (Hypericum perforatum) is an effective antidepressant can be challenged due to questionable methodology. We attempt to correct this by a meta-analysis utilizing only well-defined clinical trials. Controlled, double-blind studies using strictly defined depression criteria were analyzed by the rate of change of depression and by the number of "treatment responders." Rates of side effects and dropouts were also analyzed. Hypericum was 1.5 times more likely to result in an antidepressant response than placebo and was equivalent to tricyclic antidepressants (TCAs). The meta-analysis also showed that there was a higher dropout rate in the TCA group and that the TCAs were nearly twice as likely to cause side effects, including those more severe than hypericum. Hypericum perforatum was more effective than placebo and similar in effectiveness to low-dose TCAs in the short-term treatment of mild to moderately severe depression. However, design problems in existing studies prevent definitively concluding that St. John's wort is an effective antidepressant.

Chromosomal localization and neural distribution of voltage dependent calcium channel beta 3 subunit gene.

Voltage dependent calcium channel (VDCC) mediates the influx of free calcium ions that acts as a signal transducer. The beta 3 subunit of the VDCC regulates the activation (opening) and inactivation (closing) kinetics through phosphorylation/dephosphorylation. We isolated a genomic clone of the human VDCC beta 3 subunit from a human genomic DNA library using VDCC beta 3 cDNA as a probe. We localized VDCC beta 3 with this genomic DNA on the chromosome by fluorescent in situ hybridization, and the distribution of VDCC beta 3 in the nervous system was investigated in rats by in situ hybridization histochemistry with rat VDCC beta 3 cDNA. The gene for the VDCC beta 3 was specifically localized on human chromosome 12q13. The mRNA for the VDCC beta 3 was predominantly expressed in the nervous system. In the brain, a strong expression of VDCC beta 3 mRNA was found in the medial habenular nucleus, a high level of expression was observed in the olfactory bulb and cerebellum, and a relatively high level of VDCC beta 3 mRNA was localized in the cerebral cortex, caudate-putamen and hippocampal formation. Interestingly, this distribution pattern is very similar to that of the rbE-II, mid-low VDCC 1 subunit, and it is suspected that VDCC beta 3 and rbE-II may function together as a pair.

Accumulation of gossypol enantiomers in ovine tissues.

Tissue residue levels of gossypol enantiomers in cottonseed-fed and lethally intoxicated lambs were determined by the high-performance liquid chromatography-ultraviolet detection method. Gossypol was derivatized with (+)-2-amino-1-propanol and separated with a reversed-phase C18 column and the elution of analytes was monitored at 254 nm. The highest residue level was found in the liver tissue (318-416 ng total gossypol/mg dry tissue), and the residue of (-)-gossypol was higher than (+)-gossypol in the heart, muscle and spleen. The detection limit was 2 ng, and the detector response of gossypol-amine adducts was linear between 2 and 100 ng enantiomers.

Role of thiol agents in protecting against the toxicity of helenalin in tumor-bearing mice.

Helanalin, a sesquiterpene lactone antineoplastic agent, is toxic at therapeutic doses in murine tumors. The toxicity has been assumed to be correlated with the binding of the drug to cellular thiol groups. Studies were undertaken to increase the intracellular level of GSH in the liver, kidney and other tissues to eliminate the toxicity of helenalin in vivo. Combination of helenalin 8 mg/kg/day i.p.) with L-cysteine (100 mg/kg/day), beta-mercaptoethanolamine (20 mg/kg/day), 18-beta-glycyrrhetinic acid (15 mg/kg/day), or 4,4'-diaminodiphenylsulfone (10 mg/kg/day) afforded improvement in survival of mice bearing P-388 lymphocytic leukemia. However, other thiol-elevating agents, anti-oxidants, intracellular buffering agents, and cardiac treatment drugs were not effective. Hydrocortisone, Cortef, treatment with helenalin afforded improvement in life expectancy. Reduced glutathione (GSH) and non-protein sulfhydryl (NPS) levels were not reduced in the liver, kidney, or circulating red blood cells (rbc) by helenalin treatment. After three days treatment of mice with helenalin, GSH levels were reduced and NPS levels elevated in P-388 tumor cells. Administration of L-cysteine, beta-mercaptoethanolamine, 4,4'-diaminodiphenylsulfone, or 18-beta-glycyrrhetinic acid alone caused no alteration in liver GSH but elevated NPS levels; P388 cell GSH and NPS levels were lowered. Combination of any of these agents, after three days, with helenalin afforded increases in P-388 cell GSH and NPS levels. This data would suggest that helenalin toxicity is not related to the lowering of GSH or NPS levels in critical tissues of mice.

Anatomical and behavioral analyses of the inheritance of audiogenic seizures in the progeny of genetically epilepsy-prone and Sprague-Dawley rats.

Our previous studies have shown an increase in the number of GABAergic and total neurons in the inferior colliculus (IC) of the genetically epilepsy-prone rat (GEPR-9) as compared to the non-seizing Sprague-Dawley (SD) rat. To determine whether an increase in neuron number in the IC is genetically associated with seizure behavior, seizing and non-seizing offspring of GEPR-9 and SD progenitor strains were studied as well as offspring from backcrosses made with F1 and either GEPR-9 or SD rats. In addition, the ontogeny of seizure behavior was studied in seizing rats from these same backgrounds. The development of seizure behavior in GEPR-9s was shown to be dependent on age and the number of exposures to sound stimulus up until the age of 9 weeks. The F1 and F2 generations displayed different audiogenic seizure profiles than those of the two progenitor strains. In the F1 generation, the ratio of seizing to non-seizing rats was always greater than 3:1, and the distribution of seizure scores was similar for males and females. In addition, the off-spring from backcrosses made with F1 rats (high or low seizing) and GEPR-9s displayed maximal audiogenic response scores (ARS) of 9, a characteristic of the GEPR-9s used in this study. The results of these genetic studies indicate a polygenetic inheritance of this autosomal dominant trait of audiogenic seizure susceptibility. For the quantitative study of neuronal density in the IC, neurons were counted from cresyl violet-stained preparations from seizing and non-seizing F1 and F2 rats, backcrosses from different categories and age-matched SD rats. Statistically significant increases in the number of both small (70% increase) and medium-sized (14% increase) neurons occurred in the high seizing animals (ARS = 7-9) as compared to either the non-seizing F2 or SD rats. In addition, a significant increase in the number of small neurons (77% increase) occurred in the high seizing offspring of the F1 X GEPR-9 backcross as compared to that of the non-seizing offspring of the F1 X SD backcross. The data from 25 rats generated a 0.9 coefficient of linear correlation between ARS and the number of small neurons. The results from the anatomical studies suggest that the inheritance of audiogenic seizures appears to be closely linked to the increase in cell number. Therefore, the increase in cell number in the IC may be an important determinant of seizure behavior for GEPR-9s.

Increased numbers of neurons occur in the inferior colliculus of the young genetically epilepsy-prone rat.

To determine if the increase in the number of neurons observed in the inferior colliculus (IC) of the adult genetically epilepsy-prone rat (GEPR) as compared to the Sprague-Dawley rat was present in the young GEPRs prior to the time at which seizure activity commences, brains from both types of rats 4-10 days of age were studied. A statistically significant increase in the numbers of small neurons occurred in the IC of the young GEPR. At 4 days of age, a 55% increase in the number of small neurons was found in the GEPR as compared to the Sprague-Dawley rat and at 10 days of age this increase was 105%. The numbers of the medium and large neurons were similar in the older group of rats. These data suggest that the increase in cell number observed in the adult GEPR is not compensatory to the seizure activity, but is genetically programmed.

Elimination of adverse effects of ethoxyquin (EQ) by methionine hydroxy analog (MHA). Protective effects of EQ and MHA for bitterweed poisoning in sheep.

Dietary ethoxyquin (EQ) and methionine hydroxy analog (MHA) protected 6-8-month-old wethers from toxic doses of bitterweed (Hymenoxys odorata DC.). The EQ-MHA group received sweet feed (corn, oats, dehydrated alfalfa pellets, cane molasses and minerals), 500 g/day/sheep, supplemented with EQ and MHA (0.5% and 1.0% of feed, respectively) for 9 days prior to the poisoning with bitterweed while the MHA group received the same feed without EQ and controls received the same amount of feed with no additives. Two of 6 MHA-treated and 3 of 7 controls died whereas all 7 EQ-MHA-treated sheep survived after receiving 5 doses of bitterweed (5 X 5.5 g/kg) in 6 days. Coadministration of MHA and EQ eliminated the adverse effect of EQ; dietary EQ lowered the serum albumin, calcium, and alkaline phosphatase content while protecting the animals from bitterweed poisoning. EQ is the most promising protective agent tested for bitterweed poisoning in sheep.

Protective effects of antioxidants on bitterweed (Hymenoxys odorata DC) toxicity in sheep.

Antidotal effects of the 2 antioxidants butylated hydroxyanisole (BHA) and ethoxyquin (EQ) were evaluated in bitterweed (Hymenoxys odorata DC) toxicosis in sheep. Bitteerweed contains a toxic sesquiterpene lactone, hymenoxon, the toxicity of which is reduced by cysteine. Both BHA and EQ are known to induce hepatic glutathione production in rodents. Treatment of sheep with EQ (2.5 g/sheep/day for 9 days before poisoning) gave significant protection from toxic doses of bitterweed, but the protective effect of BHA was insignificant. Of 6 sheep given EQ in the feed, 5 survived 7 doses of bitterweed (4 g/kg/day or higher for 7 days), whereas 5 of 7 controls and 4 of 7 sheep given feed with added BHA died. The added EQ in the feed decreased the serum alkaline phosphatase activity and total protein, albumin, and calcium concentrations. Seemingly, EQ is the first protective agent with field application potential for bitterweed toxicity.

The antagonistic effect of L-cysteine in experimental hymenoxon intoxication in sheep.

L-Cysteine was given IV at dosages of 34.5, 48.8, 69.0, 97.6, or 138.0 mg/kg of body weight to groups of sheep previously treated (10.6 minutes) intraperitoneally with an LD92 of hymenoxon (12.8 mg/kg). L-Cysteine at dosages of 69.0, 97.6, and 138.0 mg/kg significantly increased survival rate 8.4-, 10-, and 12.5-fold, respectively, over that of gymenoxon-treated sheep not given L-cysteine. L-Cysteine at dosages of 34.5 and 48.8 mg/kg failed to significantly reduce mortality. Surviving hymenoxon-treated sheep given 97.6 or 138.0 mg of L-cysteine/kg showed mild or no clinical signs of toxicosis. There was no dosage of L-cysteine that significantly changed the survival time for fatally intoxicated sheep.