Perillyl alcohol mediated radiosensitization via augmentation of the Fas pathway in prostate cancer cells

BACKGROUND: The management of hormone-insensitive locally advanced prostate cancer is difficult and complex and there is an urgent need for the development of effective chemotherapeutic agents intended for combination with currently available treatment modalities. METHODS: The present paper demonstrates the effectiveness of the monoterpene perillyl alcohol (POH) as potent radiosensitizer on DU145 and PC3 cell lines by performing clonogenic survival assays, cycle analysis, and assays to detect viability, apoptosis, and Fas receptor/ligand by flow cytometry. RESULTS: POH pretreatment resulted in a dose dependent sensitization to kill cell by radiation. Furthermore, POH treatment induced a transient G(2)/M arrest, enhanced the expression of the membrane bound form of the Fas ligand and sensitized the cells to Fas mediated apoptosis. CONCLUSIONS: The unique manner of radiosensitization in addition to its low toxicity profile makes POH a promising new agent for preclinical evaluation as a potential radiosensitizer in the treatment of prostate cancer.     

Characterization of immune response in mice to plasmid DNA encoding dog zona pellucida glycoprotein-3

To investigate the immunogenicity of plasmid DNA encoding dog zona pellucida glycoprotein-3 (dZP3), the cDNA corresponding to dZP3, was cloned in mammalian expression vector VR1020 downstream of tissue plasminogen activator signal sequence under cytomegalovirus promoter (VRdZP3). In vitro transfection of COS-1 mammalian cells with VRdZP3 plasmid DNA led to its cytosolic expression. The expressed dZP3 has an apparent molecular weight of 45kDa as compared to calculated molecular weight of 38.4 kDa, suggesting possible glycosylation. Immunization of male BALB/cJ mice with VRdZP3 plasmid DNA in saline, by electroporation or adsorbed onto gold microcarriers (delivered by gene gun) generated antibody response against Escherichia coli expressed recombinant dZP3 (r-dZP3). Administration of r-dZP3 in saline following immunization with plasmid DNA led to boosting of the antibody response. Although mice immunized with gene gun exhibited highest antibody titres, the differences in the antibody titres seen by the three modes of plasmid DNA delivery were not statistically significant (P>0.05). Interestingly, female mice immunized with VRdZP3 plasmid DNA using gene gun also generated antibodies against r-dZP3. A dominant IgG1 isotype response was observed in mice immunized with VRdZP3 plasmid DNA using gene gun as compared to a mixed IgG1-IgG2a isotype response when delivered in saline or by electroporation. Immunization with VRdZP3 plasmid DNA also generated cell mediated immune response. The antibodies generated by VRdZP3 plasmid DNA recognized dog native zona pellucida. These studies for the first time, demonstrate the feasibility of generating an immune response to ZP3 by DNA vaccine and that the antibodies thus generated recognize native zona pellucida.     

Short-term impact of an acute attack of malaria on the cognitive performance of schoolchildren living in a malaria-endemic area of Sri Lanka

A prospective study was conducted from January 1998 to November 1999 in a malaria-endemic area of Sri Lanka to determine the short-term impact of an acute attack of malaria on the cognitive performance of 648 schoolchildren attending grades 1 to 5 (mostly aged 6-11 years) in 4 schools. Three groups were studied comprising children with malaria, children with non-malarial fever, and healthy controls. Cognitive performance in language and mathematics at the time of presentation and 2 weeks later was assessed. At the time of presentation, children with malaria scored significantly less in both mathematics and language than children with non-malarial fever and healthy controls. Two weeks later, the mathematics and language scores of children with malaria improved but the scores were significantly lower than the scores of children with non-malarial fever (P < 0.001) and controls (P < 0.001). Having malaria was a significant predictor of cognitive performance after controlling for other confounding factors. These findings suggest that an acute attack of uncomplicated malaria causes significant short-term impairment of cognitive performance. The impairment persists for more than 2 weeks and appears to be cumulative with repeated attacks of malaria.     

Phase I study of AG 331, a novel thymidylate synthase inhibitor, in patients with refractory solid tumors

Purpose: This was a phase I study of AG 331 to determine systemic tolerance and pharmacokinetics following single and multiple escalating intravenous doses. Methods: The study was an open-label phase I trial that was divided into two components. In phase IA (single dose), six dose levels from 12.5 to 225 mg/m² were administered to 18 patients (3 at each dose level) and serial blood samples were collected for 72 h. Upon achieving satisfactory pharmacologic parameters, the multiple dosing component (phase IB) was initiated. Six dose levels from 50 to 800 mg/m² per day were administered for 5 consecutive days to 18 patients. Pre- and postdose blood samples were obtained on days 1–4 and serial blood samples were collected over 24 h following dose 5. Nonhematologic and hepatic toxicities were assessed, serum AG 331 concentrations were measured and pharmacokinetic parameters determined. Results: Other than fatigue, no severe toxicities were encountered in phase IA. Liver toxicity was manifested by elevations in transaminase first noted at multiple doses of 200 mg/m² per day for 5 days. Fever and malaise but no myelosuppression were noted. The mean terminal t_1/2 following single doses was significantly shorter than the t_1/2 following multiple dosing (6.8 vs 9.9 h) and clearance was significantly faster following single doses than following multiple dosing (81.7 vs 30.4 1/h), but no significant difference in V_d was noted. Conclusions: The dose-related toxicity profile precludes further clinical development at this time. The pharmacokinetics of AG 331 following single and multiple doses showed significant differences. Received: 11 July 1997 / Accepted: 18 September 1998     

Quantitative study of plasticity in the auditory nuclei of chick under conditions of prenatal sound attenuation and overstimulation with species specific and music sound stimuli

Morphological effects of prenatal sound attenuation and sound overstimulation by species specific and music sounds on the brainstem auditory nuclei of chick have been evaluated quantitatively. Changes in length, volume, neuron number, size of neuronal nuclei and glial numbers of second and third order auditory nuclei, n. magnocellularis (NM) and n. laminaris (NL), were determined from thionine-stained serial sections of control and experimental groups on posthatch day 1 using stereological methods. Significant increase in volume of both auditory nuclei attributable to increase in length of nucleus, number and size of neurons, number of glia as well as neuropil was observed in response to both species specific and music overstimulation given during the critical period of development. The enhanced development of auditory nuclei in response to enriched environment prenatally indicates a positive effect of activity on neurons which may have clinical implications in addition to providing explanation for preference to auditory cues in the postnatal life. Reduction in neuron number with a small increase in proportion of cell nuclei of large size as well as an increase in glial numbers was seen in both NM and NL of the prenatally sound attenuated chicks. The increase in size of some neuronal nuclei may probably be evidence of enhanced synthesis of proteins involved in cell death or an attempt at recovery. The dissociated response of neurons and glia under sound attenuated and auditory stimulated conditions suggests that they are independently regulated by activity-dependent signals with glia also being under influence of other signals for a role in removal of dead cell debris.     

Ocular alterations in patients of alopecia areata

There is paucity of published work on ocular alterations in patients of alopecia areata (AA), especially from the Asian continent. We studied the clinical profile of 83 patients of AA and 80 sex- and age-matched controls to assess and compare the ocular changes, namely punctate opacities, cataract, intraocular tension and retinal changes. The outcome was analyzed with respect to prevalence of atopy, concomitant personal or family history of autoimmune diseases and nail changes in both the groups. The prevalence of atopy and family history of autoimmune diseases was significantly higher in the patient group. Lenticular changes were observed in 40.9% patients (including cataract in 16.9%) and 11.2% controls ( P  < 0.005). Within the patient group, lenticular involvement occurred with increased frequency in atopics ( P  = 0.034) and in the presence of family history of autoimmune diseases ( P  < 0.05). Retinal changes in the form of degenerative changes, pigmentary clumping and abnormal vascular changes were more prevalent ( P  < 0.001) in the study group. As the ocular changes were not found to correlate with the age, severity or extent of the disease, an initial ophthalmological screening of all patients is suggested.     

Pentamidine is an inhibitor of PRL phosphatases with anticancer activity

The PRL family oncogenic phosphatases are attractive targets for developing inhibitors as anticancer therapeutics given their potentially pathogenic role in human malignancies. Herein we demonstrate that pentamidine, an anti-protozoa drug with an unknown mechanism of action, is an inhibitor of PRLs with anticancer potential. Pentamidine at its therapeutic doses inhibited recombinant PRL phosphatases in vitro and inactivated ectopically expressed PRLs in NIH3T3 transfectants with an effective duration more than 24 h after a pulse cell treatment. The drug had in vitro growth-inhibitory activity against human cancer cell lines that express the endogenous PRLs. Pentamidine at a tolerable dose markedly inhibited the growth of WM9 human melanoma tumors in nude mice coincident with the induction of tumor cell necrosis and is capable of inactivating ectopically expressed PRL-2 in the cancer cells. These observations suggest the potential of pentamidine in anticancer therapies and may provide a basis for developing novel PTPase-targeted therapeutics.     

Precursor Frequency and Affinity Determine B Cell Competitive Fitness in Germinal Centers,Tested with Germline-Targeting HIV Vaccine Immunogens

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UGT2B7 promoter variant -840G>A contributes to the variability in hepatic clearance of morphine in patients with sickle cell disease

The purpose of the study was to determine if UDP-glucuronosyltransferase (UGT) 2B7 allelic variants encoding for UGT2B7, primary enzyme responsible for morphine glucuronidation contribute to the variability in the hepatic clearance of morphine in sickle cell disease (SCD). Twenty-four hour PK study of morphine and UGT2B7 variants genotyping was performed in 20 SCD patients in a steady state of health. Presence of the -840G allele (GG and GA) was associated with lower morphine metabolites/morphine AUC ratio compared with AA genotype (1.8 +/- 0.5 vs. 3.0 +/- 1.8 for M6G/M and 10.1 +/- 2.7 vs. 15.7 +/- 9.4 for M3G/M) (P = 0.03). Presence of UGT2B7 -840G allele is associated with significantly reduced glucuronidation of morphine and thus contributes to the variability in hepatic clearance of morphine in SCD.