Update in glaucoma medicinal chemistry: emerging evidence for the importance of melatonin analogues.

Glaucoma is a chronic progressive optic neuropathy, which can result in visual impairment and blindness. Elevated intraocular pressure (IOP) is currently the only modifiable risk factor. Several recent studies have shown the benefits of IOP reduction in open-angle glaucoma. Therefore, current glaucoma drugs are IOP-lowering substances such as α(2)-adrenergic agonists, ß(2)-adrenergic antagonists, carbonic anhydrase inhibitors and hypotensive lipids, which are used separately or in combination. In spite of the wide variety of antiglaucoma medicines, all therapies have several undesirable side effects. As a consequence, there are constant research attempts on the discovery of novel ocular hypotensive drugs. In the current paper, we review the latest available patents and literature for the pharmacological treatment of glaucoma, focusing on their molecular targets and/or their chemical characteristics and especially directed to melatoninergic drugs. Melatonin is a hormone secreted into the blood mainly from the pineal gland allowing the entrainment of the circadian rhythms of several biological functions. Melatonin and its analogues potently reduce IOP in rabbits, monkeys and humans. In addition, there are indications of long-term hypotensive effects and a proven neuroprotective role of melatoninergic substances. Furthermore, antidepressant and normalizing circadian rhythm actions of melatonin analogues might be beneficial for glaucoma patients. All the above mentioned facts suggest these agents as proper candidates for the glaucoma treatment. Consequently, the scientific research has given new and significant progress on the development of new, potent and selective melatonin ligands.

Real-time PCR quantification of haematopoietic chimerism after transplantation: a comparison between TaqMan and hybridization probes technologies.

This study aimed to compare the sensitivity and accuracy of two methods of quantitative real-time polymerase chain reaction (qrt-PCR), in order to determine haematopoietic chimerism (CH): single nucleotide polymorphisms using TaqMan (TM) probes and insertion/deletion polymorphisms using Hybridization (Hyb) probes. A total of 106 samples from 20 patients who underwent allogenic stem cell transplantation (n = 14) or live-donor liver transplantation (n = 6) were studied. The mean level of chimerism was 8.37% for the TM method and 7.73% in the Hyb method, which was not significantly different (P = 0.69). The Pearson correlation coefficient between the two methods was r = 0.91 (P < 0.001). The estimation of the regression line, using the Passing and Balbock method was Intercept A -0.0381 [95% confidence interval (CI) -0.1265 to 0.0296) and Slope B: 1.04609(95% CI 0.9349-1.161). Bland-Altman data showed that the standard deviations, which differed between the two methods (%Hyb-%TM), were 0.98 and -1.28. The accuracy and sensitivity of qrt-PCR chimerism is independent of the method used if the optimization is adequate and satisfies the criteria for adequate study. Real-time PCR, independent of the method adopted, is a very good tool for study levels of CH.

Comparison of the MagNA pure LC automated system and the RiboPure-Blood RNA manual method for RNA extraction from multiple myeloma bone marrow samples conserved in an RNA stabilizer.

A total of 62 frozen bone marrow specimens conserved in RNA later (Ambion) were processed using two different extraction methods, the MagNA Pure LC system (MAG; Roche) and the manual RiboPure-Blood RNA method (RIBO; Ambion); Beta glucoronidase RNA (GUS) was amplified by LightCycler PCR to evaluate the quality of both extraction procedures. Less than 1000 GUS copies/ml was detected in 26 of 62 specimens (41.94%) processed by MAG and in five of 62 specimens (8.06%) processed by RIBO. Moreover, RNA recovery from the 62 specimens by MAG is, on average, 2.91 cycle threshold-fold higher than RIBO (P = 0.0008). Furthermore, we compared the extraction times and reagent costs of both methods. In conclusion, RNA extraction using MAG is faster to process 32 samples and less expensive than RIBO but it is not sensitive enough to be employed for research purpose in our laboratory.

Clomiphene citrate and human chorionic gonadotropin in the treatment of anovulatory infertility.

Fifty-one patients with proven anovulatory infertility were treated for four months. For two months each, a dose regimen of 50 mg of clomiphene citrate daily for five days and one of 150 mg daily for five days was prescribed. During one month of each of these two regimens, each patient also received 5000 IU of human chorionic gonadotropin intramuscularly. Thirty-three of the 51 patients ovulated at least once during the four months of treatment; the ovulation rate was 64.7%. Sixteen pregnancies occurred during the treatment, for a pregnancy rate of 31.4%. Human chorionic gonadotropin did not appear to increase the ovulation rate.

Analysis of variables in treatment of anovulation with human gonadotrophins.

Twenty seven women with anovulation were given 121 courses of gonadotrophins monitored by excretion of estrogen and pregnanediol. The FSH, ovulating dosage of HCG, time interval between FSH and HCG and subsequent supporting doses of HCG were varied in a statistical design. None significantly affected ovulation or pregnancy rates. The preparation with an FSH:LH ratio of 1 required a lower dosage to initiate an estrogen response than that with a ratio of 5. After adjustment it still produced a faster rise in estrogen excretion. The amount of estrogen also varied with the time interval between FSH and HCG, the ovulating dose of HCG and supporting doses of HCG. The amount of pregnanediol varied with the ovulating dose of HCG and the length of the luteal phase varied with the ovulating dose of HCG and supporting injections of HCG. The dosage of FSH needed to initiate a response increased on average by 8% from one course to the next. Simplicity, economy and risk of multiple pregnancy are discussed in relation to these variables.