Therapeutic applications of melatonin and related compounds

Increasing knowledge of the pharmacological effects of melatonin has suggested various possible therapeutic applications for the hormone. Because, as a natural substance, melatonin cannot be patented, melatonin-related compounds have been synthesized by industrial groups. The scope of such compounds is also to specifically target the recently discovered melatonin receptor subtypes. The sleep-inducing properties of melatonin are disputed, but are distinct from those of benzodiazepines. The observed effects on sleep latency or sleep efficiency, which remain to be confirmed, could be accounted for by the effects of melatonin on core body temperature and on circadian rhythms. There is also an urgent need for safety data, both in animals and in humans, particularly when long-term use is envisaged.     

Therapeutic comparison of thiol compounds in severe paracetamol poisoning

Twelve patients with toxic blood concentrations of paracetamol were treated with either cysteamine or amino-acid solution. None of the patients developed severe liver damage, although transient mild biochemical abnormalities developed in three. None of the patients treated with amino-acid solution had side effects due to therapy, whereas all those treated with cysteamine did. It is recommended that amino-acid solutions be used as a temporary measure in patients suspected of massive paracetamol overdose while awaiting estimation of blood paracetamol concentration.     

A novel cell-based assay for the evaluation of anti-ras compounds

In order to identify drugs active against mutated ras oncogenes we have developed an in vitro assay employing two clones of the human fibrosarcoma cell-line, HT1080 which carries an N-ras gene mutated at codon 61. Clone, HT1080scc2, retains the transformed phenotype of the parental line, whilst the other, HT1081c, is a morphologically flat, non-tumourigenic, revertant with under-representation of the chromosome carrying the transforming N-ras allele. The clear implication of mutant ras in maintaining the transformed nature of HT1080scc2 was confirmed when these cells were microinjected with the pan ras neutralising antibody Y13-259, which resulted in the morphological detransformation of these cells to a phenotype resembling that of the HT10801c clone. A number of known anti-cancer drugs with modes of action unrelated to ras function were found to be equipotent against both clones. However, when compounds chosen on the grounds of their potential selective cytotoxic or differentiating activity were tested some interesting results were obtained. Thus 8-bromo cAMP affected some morphological detransformation of HT1080scc2 cells and reduced their colony forming potential. The IMP-dehydrogenase inhibitors, tiazafurin and mycophenolic acid also flattened the morphology of the transformed clone. Fumagillin, an antibiotic reported to exhibit selective activity against ras transformed cells showed very marked and selective cytostatic effects against HT1080scc2 cells with IC50 values as low as 1 x 10(-11) M.     

Comparative efficacy evaluation of dicationic carbazole compounds, nitazoxanide, and paromomycin against Cryptosporidium parvum infections in a neonatal mouse model

The efficacies of dicationic carbazole compounds, nitazoxanide (NTZ), and paromomycin were evaluated against the AUCp1 isolate of Cryptosporidium parvum by using a neonatal mouse model. Compounds were solubilized or suspended in deionized water and administered orally by gavage to neonatal mice at a constant dose rate on days 0 to 5 (treatment started on day 0). Dose rates varied for individual carbazole compounds but ranged from 0.65 to 20 mg/kg of body weight. NTZ was tested at 100 and 150 mg/kg, and paromomycin was tested at 50 mg/kg. Efficacies were determined by comparing numbers of oocysts present in treated versus control mice at necropsy examination on day 6. Demonstrable efficacy was observed for several carbazole compounds, based on significant reductions in the numbers of oocysts recovered from treated mice versus control mice. Compounds 1, 7, and 10 (19.0 mg/kg) reduced oocyst passage in treated mice to less than 5% of that in control mice. Treatment with compounds 6, 8, and 9 (17.0 mg/kg) resulted in reductions of oocyst output to less than 10% of that in controls. Although they were not comparable in efficacy to compounds 1, 6, 7, 8, 9, and 10, treatment with other carbazole compounds resulted in statistically significant reductions in oocyst output in treated versus control mice. Compound 1 retained efficacy resulted in reduction of oocyst output to approximately 6% of that in controls when the dose was reduced to 5 mg/kg. Further reductions in the dose rate resulted in considerable reductions in anticryposporidial activity. Likewise, the efficacies of compounds 9 and 10 were reduced substantially when the doses were lowered to one-half the screening dose. Paromomycin yielded excellent activity (reduction of oocyst output to <2% of that in controls) at a dose of 50 mg/kg. NTZ yielded moderate efficacy as powder and injectable formulations administered at 100 mg/kg orally (reduction of oocyst output to 42 and 26% of that in controls, respectively). Oral administration of the injectable formulation of NTZ at a dose of 150 mg/kg resulted in improved efficacy (oocyst output, <5% of that in controls).     

Therapeutic trials in the murine model of hereditary tyrosinaemia type I: a progress report

We have studied a knockout mouse with fumarylacetoacetate hydrolase (FAH) deficiency as a model of human hereditary tyrosinaemia type (I (HT1). These mice have a phenotype very similar to the human disease, which is characterized by acute hepatic failure, renal tubular disease and hepatocarcinoma. We have previously reported on the efficacy of 2-(2-nitro-4-trifluoromethylbenzyol)-1,3-cyclohexanedione (NTBC) in preventing acute liver disease in HT1 mice. Here we present a progress report on long-term follow up (> 1 year) of high-dose NTBC therapy in combination with tyrosine restriction. In vivo retroviral gene therapy was also effective in abolishing the acute liver failure of HT1. Retrovirally treated mice remained completely healthy and active for 12 months after retroviral gene transfer. However, hepatocarcinoma developed in 2/3 treated animals after 1 year. Southern blot analysis showed that the tumours did not arise from retrovirally transduced hepatocytes but from non-corrected FAH-deficient cells. These results highlight the extreme danger for tumour formation in HT1 and indicate the need for improved gene therapy that leads to the elimination of endogenous FAH-deficient liver cells.     

Effects of immunopharmacological compounds in the Lyme arthritis model using normal and SCID mice

The murine Lyme borreliosis causes a special type of arthritis whose development appears to be controlled by a functioning immune system. Immunocompetent C3H and severe combined immunodeficient (SCID) mice were infected with Borrelia burgdorferi (strain SH-2-82) to induce experimental Lyme disease. Expression of clinical symptoms was mild to very moderate in the C3H but more rapidly developing and severe in the SCID mouse. Various pharmacological compounds, such as anti-inflammatory and immunosuppressive drugs, monoclonal antibodies and other miscellaneous agents, were investigated for profiling their effects in this model in both mouse strains. Several disease parameters were assessed, in particular paw swelling. The use of these various compounds provided further evidence that experimental borreliosis in mice represents a special type of arthritis which has no autoimmune basis and which requires productive infection with Borrelia burgdorferi. In addition, when comparing these results with those obtained in other mainly immune driven arthritis models commonly used in inflammation research, it is concluded that this arthritis model is not suitable for the therapeutic assessment of antiinflammatory agents.     

Development of a model for evaluation of microbial cross-contamination in the kitchen

Foods can become contaminated with pathogenic microorganisms from hands, the cutting board, and knives during preparation in the kitchen. A laboratory model was developed to determine occurrence of cross-contamination and efficacy of decontamination procedures in kitchen food-handling practices. Enterobacter aerogenes B199A, an indicator bacterium with attachment characteristics similar to that of Salmonella spp., was used. Chicken meat with skin inoculated with 10(6) CFU of E. aerogenes B199A/g was cut into small pieces on a sterile cutting board. The extent of cross-contamination occurring from meat to the cutting board and from the cutting board to vegetables (lettuce and cucumbers) subsequently cut on the board was determined. Swab samples from the cutting board, hand washings, and lettuce and cucumber samples revealed that approximately 10(5) CFU of E. aerogenes/cm2 were transferred to the board and hands and approximately 10(3) to 10(4) CFU of E. aerogenes/g to the lettuce and cucumbers. The surfaces of the cutting board and hands were treated with antibacterial agents after cutting the meat, and counts of E. aerogenes on the cutting board and vegetables (lettuce and cucumbers) were determined. Results revealed that use of the disinfectant reduced the population of E. aerogenes to almost nondetectable levels on the cutting boards. The average counts after treatment were < 20 CFU/g of vegetable and ranged from < 20 to 200 CFU per cm2 or g on the cutting board and subsequently on the vegetables. These results indicate that bacteria with attachment characteristics similar to Salmonella spp. can be readily transferred to cutting boards during food preparation and then cross-contaminate fresh vegetables if the boards are not cleaned. Application of a kitchen disinfectant can greatly reduce bacterial contamination on cutting boards.     

模糊综合模型在采空区稳定性评价中的应用

采空区稳定性影响因素众多且相互作用关系复杂,基于模糊数学理论,对采空区稳定性等级进行定量评价。从岩体质量、采场结构特征和工程环境等3方面入手,将采空区稳定性影响因素细分为3大类共13个指标,构建采空区稳定性评价指标体系。通过构建指标因素集、权重集和评价集,建立采空区稳定性模糊综合评价模型。以某地下开采金矿为对象,应用模糊综合模型对其-200 m中段采空区稳定性分级。对象采空区中有5个被划分为Ⅲ级(中等稳定),1个为Ⅳ级(不稳定)。评价结果与现场实际情况相符,验证了模糊综合评价模型的有效性。     

Issues in the management of human papillomavirus genital disease

Human papillomavirus (HPV) infects epithelial cells at a variety of anatomic sites. Of the more than 80 types of this virus, approximately one third infect genital tissue. Genital infection can be clinical (warts), subclinical (abnormal Papanicolaou [Pap] smears or lesions visible only with magnification and application of acetic acid) or latent (infection with apparently normal tissue). Recent research has revealed a clear association between genital warts and HPV types 6 and 11, and between cervical cancer and HPV types 16, 18 and others. Women with genital warts who have normal Pap smears do not need colposcopy. All available therapies are suboptimal because HPV is not eradicated by treatment. However, the active disease expression can be eliminated by treatment. The approach to treatment will depend on each patient's personal characteristics and preferences.     

Selection of a statistical model for the evaluation of the reliability of the results of toxicological analyses. II. Selection of our statistical model for the evaluation

Part II presents a statistical model devised by the authors for evaluating toxicological analyses results. The model includes: 1. Establishment of a reference value, basing on our own measurements taken by two independent analytical methods. 2. Selection of laboratories -- basing on the deviation of the obtained values from reference ones. 3. On consideration of variance analysis, t-student's test and differences test, subsequent quality controls and particular laboratories have been evaluated.