Gossypol acetic acid (GAA) has been shown to have male antifertility effects, but there are pronounced differences among animal species. In the search of endogenous effector molecules, which interfere with the functions of GAA, we have studied the in vitro effect of various amino acids on the inhibition of the purified LDH-X by GAA. Histidine, cysteine and glycine were shown to block the effect of GAA. The effects of these amino acids were concentration dependent. Histidine and glycine protection was found to be complex type in which both the Km and Vmax were decreased compared to control. Arginine, glutamic acid, phenylalanine and valine were found to be ineffective against the inhibitory action of GAA.
Amino Acids/pharmacology* ; Animal ; Enzyme Inhibitors/pharmacology* ; Goats ; Gossypol/pharmacology ; Gossypol/analogs & derivatives* ; Isoenzymes ; Lactate Dehydrogenase/antagonists & inhibitors* ; Male ; Spermatocidal Agents/pharmacology* ; Testis/enzymology

BACKGROUND: In this report, we present the annual data of the intensive care unit (ICU) module of the Korean Nosocomial Infections Surveillance System (KONIS) from July 2008 through June 2009. METHODS: We performed a prospective surveillance of nosocomial urinary tract infections (UTI), bloodstream infections (BSI), and pneumonia (PNEU) at 101 ICUs in 57 hospitals using KONIS. Nosocomial infection (NI) rates were calculated as the numbers of infections per 1,000 patient-days or device-days. RESULTS: We identified 3,287 NIs during the study period: 1,787 UTIs (1,772 cases were urinary catheter-associated), 917 BSIs (797 were central line-associated), and 583 PNEUs (335 were ventilator-associated). The rate of urinary catheter-associated UTIs was 4.80 cases per 1,000 device-days (95% confidence interval, 4.58-5.03) and urinary catheter utilization ratio was 0.85 (0.849-0.851). Although the urinary catheter utilization ratios were lower in the hospitals with 400-699 beds than in the hospitals with 700-899 beds and more than 900 beds, the rates of urinary catheter-associated UTIs were higher in hospitals with 400-699 beds than in the larger ones. The rate of central line-associated BSIs was 3.27 (3.05-3.51) and the utilization ratio was 0.56 (0.559-0.561). The rate of ventilator-associated PNEUs was 1.86 (1.67-2.07) and the utilization ratio was 0.41 (0.409-0.411). The rate of ventilator-associated PNEUs was lower in July 2008-June 2009 than in July 2007-June 2008 and July 2006-December 2006. CONCLUSION: It appears that the KONIS influences the reduction in the rate of device-associated infections, especially ventilator-associated PNEU; therefore, ongoing targeted surveillance and infection control strategies are needed to control device-associated infections.
Cross Infection ; Gossypol ; Infection Control ; Critical Care ; Intensive Care Units ; Pneumonia ; Urinary Catheters ; Urinary Tract Infections

OBJECTIVETo study the effect of gossypol on the expression of connexin 43 (CX43) in Sertoli cells.

METHODSTM4 Sertoli cells were cultured and treated with gossypol at the concentrations of 1.25, 2.5, 5 and 10 micromol/L for 6, 12, 24 and 48 hours. The cytotoxicity of gossypol was assessed by CCK-8 assay, and the expression of CX43 in the normal TM4 Sertoli cells and in those treated with different concentrations of gossypol for different times was detected by RT-PCR and immunofluorescence analysis.

RESULTSSemiquantitative RT-PCR and immunofluorescence analysis showed the expression of CX43 in the normal TM4 cells. At 24 hours of exposure to gossypol, the expression began to decrease gradually with the prolonging of time and the increasing concentration of gossypol (P < 0.05).

CONCLUSIONGossypol reduces the expression of CX43 in TM4 Sertoli cells, which might underlie the mechanism of its antifertility action.

Cells, Cultured ; Connexin 43 ; metabolism ; Gossypol ; toxicity ; Humans ; Male ; Sertoli Cells ; drug effects ; metabolism

OBJECTIVETo establish a method for the determination of gossypol in cotton root bark.

METHODSamples were extracted with acetone, and determined on a C18 column with the mobile phase (Acetonitrile-0.2% phosphoric acid, 85:15) and UV-235 nm detector.

RESULTThe recovery rate was 97.6%, RSD 1.59% (n = 5).

CONCLUSIONThis method can be used for the determination of gossypol in cotton root bark and the content of gossypol in three different species has been determined.

Chromatography, High Pressure Liquid ; Gossypium ; chemistry ; Gossypol ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVEThis paper aims to study the effects of gossypol acetic acid (GAA) on proliferation and methylation level of human MutL homologue 1 (hMLH1) gene in human tongue cancer cell line Tca8113.

METHODSThe MTT assay was used to determine the effects of the acid on the proliferation inhibition in Tca8113 cells treated with different GAA concentrations. Nested methylation-specific polymerase chain reaction (nMSP) was used to detect the change in the methylation level of hMLH1 after 48 and 72 h with 30 and 15 micro mol L(-1) GAA treatment.

RESULTSMTT assay results showed the growth and proliferation inhibition of Tca8113 cells in the experimental GAA group after 24 h to 72 h of GAA treatment. The nMSP results indicated that the average optical density of hMLH1 in the Tca8113 cells significantly changed after the GAA treatment (30 micro mol L(-1) GAA for 48 h and 15 micro mol L(-1) for 72 h) (P<0.05) compared with that of the control group.

CONCLUSIONGAA does not only inhibit Tca8113 proliferation but also has a demethylation effect on the hMLH1 gene. These phenomena may be part of an underlying tumor-suppression mechanism of GAA.

Cell Line ; Cell Line, Tumor ; Cell Proliferation ; Gossypol ; analogs & derivatives ; Humans ; Methylation ; Tongue Neoplasms

OBJECTIVE: To study the toxic effects of short-term exposure to gossypol on the testis and kidney in mice and whether these effects are reversible. METHODS: Twenty 7 to 8-week-old male mice were randomized into blank control group, solvent control group, gossypol treatment group and drug withdrawal group. In the former 3 groups, the mice were subjected to daily intragastric administration of 0.3 mL of purified water, 1% sodium carboxymethylcellulose solution, and 30 mg/mL gossypol solution for 14 days, respectively; In the drug withdrawal group, the mice were treated with gossypol solution in the same manner for 14 days followed by treatment with purified water for another 14 days. After the last administration, the mice were euthanized and tissue samples were collected. The testicular tissue was weighed and observed microscopically with HE and PAS staining; the kidney tissue was stained with HE and examined for mitochondrial ATPase activity. RESULTS: Compared with those in the control group, the mice with gossypol exposure showed reduced testicular seminiferous epithelial cells with rounded seminiferous tubules, enlarged space between the seminiferous tubules, interstitium atrophy of the testis, and incomplete differentiation of the spermatogonia. The gossypol-treated mice also presented with complete, non-elongated spermatids, a large number of cells in the state of round spermatids, and negativity for acrosome PAS reaction; diffuse renal mesangial cell hyperplasia, increased mesangial matrix, and adhesion of the mesangium to the wall of the renal capsule were observed, with significantly shrinkage or even absence of the lumens of the renal capsules and reduced kidney mitochondrial ATPase activity. Compared with the gossypol-treated mice, the mice in the drug withdrawal group showed obvious recovery of morphologies of the testis and the kidney, acrosome PAS reaction and mitochondrial ATPase activity. CONCLUSIONS Shortterm treatment with gossypol can cause reproductive toxicity and nephrotoxicity in mice, but these toxic effects can be reversed after drug withdrawal.
Mice ; Male ; Animals ; Gossypol/toxicity* ; Testis ; Seminiferous Tubules ; Spermatids ; Spermatogenesis ; Adenosine Triphosphatases/pharmacology*

The objective of this work was to investigate the effect of six individual strains of fungi on the reduction of gossypol levels and nutritional value during solid substrate fermentation of cottonseed meal (CSM). Six groups of disinfected CSM substrate were incubated for 48 h after inoculation with either of the fungi C. capsuligena ZD-1, C. tropicalis ZD-3, S. cerevisae ZD-5, A. terricola ZD-6, A. oryzae ZD-7, or A. niger ZD-8. One not inoculated group (substrate) was used as a control. Levels of initial and final free gossypol (FG), crude protein (CP), amino acids (AA) and in vitro digestibility were assayed. The experiment was done in triplicate. The experimental results indicated that microbial fermentation could greatly decrease (P<0.05) FG levels in CSM. The detoxification efficiency differed between the species of microorganisms applied. From the perspective of reducing CSM potential toxicity, C. tropicalis ZD-3 was most successful followed by S. cerevisae ZD-5 and A. niger ZD8. They could reduce FG levels of CSM to 29.8, 63.07 and 81.50 mg/kg based on DM (dry matter), respectively, and their detoxification rate were 94.57%, 88.51% and 85.16%, respectively. If crude protein, amino acids content and their in vitro digestibility were also taken into account, A. niger ZD-8 may be the best choice. The CP content of CSM substrate fermented by C. tropicalis ZD-3 and A. niger ZD-8 were improved by 10.76% and 22.24%; the TAA (total amino acids) contents were increased by 7.06% and 11.46%, and the EAA (essential amino acids) were raised by 7.77% and 12.64%, respectively. Especially, the levels of methionine, lysine and threonine were improved greatly (P<0.05). The in vitro CP digestibility of CSM fermented by C. tropicalis ZD-3 and A. niger ZD-8 was improved by 13.42% and 18.22%, the TAA were increased by 17.75% and 22.88%, and the EAA by 16.61% and 21.01%, respectively. In addition, the in vitro digestibility of methionine, lysine and threonine was also improved greatly (P<0.05).
Amino Acids ; analysis ; Cottonseed Oil ; chemistry ; metabolism ; Digestion ; Fermentation ; Fungi ; metabolism ; Gossypol ; analysis ; Nutritive Value ; Plant Proteins ; analysis

We found that non-small-cell lung cancer (NSCLC) cells express high levels of multiple aldehyde dehydrogenase (ALDH) isoforms via an informatics analysis of metabolic enzymes in NSCLC and immunohistochemical staining of NSCLC clinical tumor samples. Using a multiple reaction-monitoring mass spectrometry analysis, we found that multiple ALDH isozymes were generally abundant in NSCLC cells compared with their levels in normal IMR-90 human lung cells. As a result of the catalytic reaction mediated by ALDH, NADH is produced as a by-product from the conversion of aldehyde to carboxylic acid. We hypothesized that the NADH produced by ALDH may be a reliable energy source for ATP production in NSCLC. This study revealed that NADH production by ALDH contributes significantly to ATP production in NSCLC. Furthermore, gossypol, a pan-ALDH inhibitor, markedly reduced the level of ATP. Gossypol combined with phenformin synergistically reduced the ATP levels, which efficiently induced cell death following cell cycle arrest.
Adenosine Triphosphate ; Aldehyde Dehydrogenase* ; Cell Cycle Checkpoints ; Cell Death ; Energy Metabolism* ; Gossypol ; Humans ; Informatics ; Isoenzymes ; Lung ; Lung Neoplasms ; Mass Spectrometry ; NAD ; Phenformin ; Protein Isoforms

To investigate the effects of gossypol acetate on apoptosis in primary cultured cells from patients with acute lymphoblastic leukemia (ALL) and chronic lymphocytic leukemia (CLL) and its synergistic effect with dexamethasone. The apoptosis-inducing effect of gossypol acetate on primary cultured leukemia cells was analyzed by flow cytometry (FCM). The effect of gossypol acetate on survival rates of Raji cells and mononuclear cells (MNC) from normal bone marrow were evaluated by MTT assay. After co-treatment with gossypol acetate and dexamethasone, the apoptosis rate of Raji cells was detected by FCM. The results showed that gossypol acetate was able to induce apoptosis in primary cultured ALL cells at concentrations of ≥ 5 µmol/L. The effect was concentration and time dependent. Apoptosis-inducing concentration in CLL cells was higher than that in ALL cells. After exposing to 50 µmol/L gossypol acetate for 48 h, the apoptosis rate of ALL and CLL cells were (90.4 ± 6.2)% and (51.7 ± 10.3)% separately. No major growth inhibitory effect was observed in MNC from normal bone marrow when they were exposed to gossypol acetate at concentrations lower than 10 µmol/L. After exposing for 48 and 72 h, the IC(50) of gossypol acetate for MNC from normal bone marrow was 7.1 and 9.1 times as much as the IC(50) of Raji cells. Co-treatment with 10 µmol/L gossypol acetate and dexamethasone remarkably increased the apoptosis rate of Raji cells. It is concluded that the gossypol acetate has apoptosis-inducing activity in primary cultured leukemia cells from patients diagnosed as ALL and CLL in vitro. The inhibitory effect of gossypol acetate on MNC from normal bone marrow is less prominent than that on Raji cells. Co-treatment with gossypol acetate and dexamethasone notably amplified the pro-apoptosis activity of the latter in Raji cells.
Apoptosis ; drug effects ; Cell Line ; Dexamethasone ; pharmacology ; Gossypol ; analogs & derivatives ; pharmacology ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; pathology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; pathology ; Tumor Cells, Cultured

Gossypol is found to induce an inhibition of fertility in various animals such as rat, dog and monkey. This study was undertaken to determine the exact site of the action of gossypol acetic acid at the testicular level concomitant with antifertility activity at various dose and time intervals in male hamsters. This study consisted of three groups of hamsters according to the dose of GAA with control group. After 12 weeks of treatment at the dose of 5mg/kg/day and 10mg/kg/day a rate of increase in body weight were statistically lower than that of control group. In contrary at the dose of 20mg/kg/day there was a tendency to decrease in body weight. Male hamsters became completely sterile after 8 weeks of treatment with 20mg/kg/day. A regaining of fertility was observed after the cessation of treatment in 5 male hamsters that lost fertility. In the group treated with 10mg/kg/day of GAA, the fertility rate of female at 4,8 and 12 weeks of treatment were reduced 54.5%, 37.5% and 35.7 % respectively whereas those of female in 5 mg/kg/day group were 91.7%, 69.2% and 71.4% respectively. The number of delivered hamsters also decreased with the increase in dosage and duration of treatment. A decrease of motility was observed and followed by reduced sperm population. It was not until 4 weeks after administration with 20mg/kg/day of GAA that a reduction of sperm population was statistically significant. Azoospermia was found as early as 8 weeks after treatment with 20mg/ke/day of GAA. ultrastructurally initial detectable damages were observed in mitochondrial sheath of epididymal sperm and the damage was progressed in order of early spermatid, primary and secondary spermatocyte. spermatogonia. Distinct ultrastructural changes were seen in mitochondria of germ cell. In Sertoli cell cytoplasmic vacuolization was noted and mitochondrial damage was observed in late phase. An aggregation of mitochondria in spermatogonia was observed after treatment with 20mg/kg/day of GAA for 12 weeks. Leydig cell was not altered significantly in this study. The results of this study confirmed antifertility effects of GAA in male hamster and suggested that a spermatogonia may be damaged by chronic effects of gossypol treatment.
Acetic Acid* ; Animals ; Azoospermia ; Birth Rate ; Body Weight ; Cricetinae* ; Cytoplasm ; Dogs ; Female ; Fertility* ; Germ Cells ; Gossypol* ; Haplorhini ; Humans ; Infertility ; Male* ; Mitochondria ; Rats ; Spermatids ; Spermatocytes ; Spermatogonia ; Spermatozoa ; Testis* ; Withholding Treatment