DDEC: Dragon database of genes implicated in esophageal cancer

Background  

Esophageal cancer ranks eighth in order of cancer occurrence. Its lethality primarily stems from inability to detect the disease during the early organ-confined stage and the lack of effective therapies for advanced-stage disease. Moreover, the understanding of molecular processes involved in esophageal cancer is not complete, hampering the development of efficient diagnostics and therapy. Efforts made by the scientific community to improve the survival rate of esophageal cancer have resulted in a wealth of scattered information that is difficult to find and not easily amendable to data-mining. To reduce this gap and to complement available cancer related bioinformatic resources, we have developed a comprehensive database (Dragon Database of Genes Implicated in Esophageal Cancer) with esophageal cancer related information, as an integrated knowledge database aimed at representing a gateway to esophageal cancer related data.     

Presence of calcitonin-like peptide in rat milk: possible physiological role in regulation of neonatal prolactin secretion

Previous results have shown that salmon calcitonin (sCT), a peptide in rat brain and pituitary gland, inhibits basal and TRH-stimulated PRL release and reduces PRL mRNA levels in isolated anterior pituitary cells of adult rats in culture. Rat milk contains a variety of neuropeptides and hormones, some of which are absorbed in bioactive form to exert endocrine influences in the developing offspring. The present studies were undertaken to investigate whether a CT-like peptide is present in rat milk. Circulating PRL levels in neonatal rats are low, and there is an abrupt increase in the basal secretion of this hormone at weaning. A second objective was to examine whether CT plays a role in the regulation of PRL secretion in neonatal animals. A sensitive and specific RIA for sCT was developed and used to assay rat milk on various days of lactation for sCT-like immunoreactivity. sCT-like activity was present in the water-soluble (infranatant) fraction of milk throughout lactation in concentrations as high as 1589 pg/ml. There were no statistically significant differences in immunoreactive levels of the peptide in milk samples from different days of lactation. sCT-like immunoreactivity in rat milk infranatant coeluted with synthetic sCT on reverse phase HPLC, and these HPLC fractions inhibited basal PRL release when added to cultures of anterior pituitary cells. This inhibition of PRL release by the sCT-immunoreactive HPLC fractions was comparable to that exerted by equivalent concentrations of synthetic sCT. Newborn rats were injected sc with 10 microliters normal rabbit serum or anti-sCT serum from the day of birth until postpartum day 10. The rats were killed on day 11, and their sera were analyzed for PRL. Anti-sCT-injected rats showed a significant increase in serum PRL levels compared to those in untreated or normal serum-treated rats. These results demonstrate that a CT-like peptide, which is a potent inhibitor of PRL release, is present in rat milk throughout lactation, and that passive immunization with a highly specific anti-sCT serum leads to an increase in serum PRL levels in neonatal rats. CT, possibly of milk origin, may be a physiologically relevant PRL-inhibiting factor during the neonatal period.     

Dynamic photoacoustic imaging of neurovascular coupling in salivary glands

The purpose of this study was to apply photoacoustic imaging (PAI), a relatively new imaging method, to non-invasively map neurovascular dynamics in salivary glands. Dynamic PAI with co-registered ultrasound (US) was performed in mice to monitor salivary gland hemodynamics in response to exogenous muscarinic receptor stimulation (pilocarpine) and blockade (atropine). Pilocarpine increased salivary gland oxygen saturation (%sO₂) within minutes after administration, which was abrogated by atropine. A significant correlation was observed between change in %sO₂ measured by PAI and saliva secretion. PAI is a novel imaging method that can be used for functional assessment of neurovascular dynamics in salivary glands.     

Metabolism and elimination of rhodamine 123 in the rat

Summary Little is known of the pharmacology of rhodamine 123 (RH-123), an agent reported to have carcinoma-selective experimental antitumor activity. Accordingly, using a high-performance liquid chromatographic assay system with fluorescence detection, we examined the plasma decay and the biliary and urinary elimination of parent drug and metabolites in female Sprague-Dawley rats receiving RH-123 at an intravenous dose (5 mg/kg) equivalent to the therapeutic dose used in murine tumor models. Following drug administration to unconscious animals, plasma levels of drug-associated fluorescence fell in a triphasic manner (t_1/2, 15 min; t_1/2, 1 h; t_1/2, 4.7 h). In plasma, unchanged drug predominated but lower levels of the deacylated metabolite rhodamine 110 (RH-110) and two unknowns were also detectable throughout the study. Drug fluorescence was recovered extensively in both urine and bile. In unconscious animals with ureteral cannulae, urinary excretion (11.4% of the dose in 6 h) occurred predominantly as unchanged RH-123 (97% of the total), with low levels of RH-110 (2.4%) and two unknowns (<0.6% combined) also being present. Similarly dosed conscious animals (without surgical intervention) housed in metabolic cages showed a comparable pattern of urinary excretion, with 11.9% of the drug dose being recovered in 6 h and 21.9%, by 48 h. Biliary drug elimination accounted for 8% of the delivered dose in 6 h in unconscious animals and for 11% by 36 h in conscious animals fitted with biliary cannulae. In contrast to urinary excretion, in which unchanged drug predominated, only 50% of the fluorescence recovered in bile was attributable to RH-123. The remainder was due to a number of products that were detectable throughout the study. Of these, one present at significant levels was identified as a glucuronide conjugate of RH-123, based on the liberation of parent drug when the purified metabolite was incubated with -glucuronidase or hydrolyzed with 1 N hydrochloric acid. Further studies with a radiolabeled form of RH-123 are necessary to establish the identity of the remaining unknowns disclosed in this work.This work was supported in part by research grants CA 44890 (T.W.S.) and CA 37082 (M.I.) from the National Cancer Institute, National Institutes of Health, United States Public Health Service     

Development and Validation of a Precise,Single HPLC Method for the Determination of Tolperisone Impurities in API and Pharmaceutical Dosage Forms

A novel, sensitive, stability-indicating HPLC method has been developed for the quantitative estimation of Tolperisone-related impurities in both bulk drugs and pharmaceutical dosage forms. Effective chromatographic separation was achieved on a C18 stationary phase with a simple mobile phase combination delivered in a simple gradient programme, and quantitation was by ultraviolet detection at 254 nm. The mobile phase consisted of a buffer and acetonitrile delivered at a flow rate 1.0 ml/min. The buffer consisted of 0.01 M potassium dihydrogen phosphate with the pH adjusted to 8.0 by using diethylamine. In the developed HPLC method, the resolution between Tolperisone and its four potential impurities was found to be greater than 2.0. Regression analysis showed an R value (correlation coefficient) of greater than 0.999 for the Tolperisone impurities. This method was capable of detecting all four impurities of Tolperisone at a level of 0.19 μg/mL with respect to the test concentration of 1000 μg/mL for a 10 µl injection volume. The tablets were subjected to the stress conditions of hydrolysis, oxidation, photolysis, and thermal degradation. Considerable degradation was found to occur in base hydrolysis, water hydrolysis, and oxidation. The stress samples were assayed against a qualified reference standard and the mass balance was found to be close to 100%. The established method was validated and found to be linear, accurate, precise, specific, robust, and rugged.     

Cervical intraepithelial neoplasia and human papilloma virus infection.

Evidence of human papilloma virus (HPV) infection was sought in 50 patients with cervical intraepithelial neoplasia (CIN) and 50 controls. Cytologic, colposcopic and histopathological examinations were carried out in each one of them. Cytological evidence of HPV infection was found in 13 cases and two controls. Colposcopic evidence was found in 33 (66%) cases and 17 (34%) controls and histopathological evidence in 35 (70%) patients and 14 (28%) controls. These differences were statistically significant. Colposcopy proved to be a good method of diagnosing HPV infection with a sensitivity of 86% and specificity of 80% compared to histopathologic diagnosis.     

Combination versus sequential doxorubicin and docetaxel as primary chemotherapy for breast cancer: A randomized pilot trial of the Hoosier Oncology Group.

PURPOSE: To evaluate the efficacy and toxicity of combination and sequential dose-dense chemotherapy with doxorubicin and docetaxel (Taxotere; Rh?ne-Poulenc Rorer, Collegeville, PA) as primary chemotherapy of breast cancer. PATIENTS AND METHODS: Patients with newly diagnosed stage II or noninflammatory stage III breast cancer were randomly assigned to receive the same total doses of doxorubicin and docetaxel over a 12-week period before definitive surgery. Patients in arm A received sequential therapy with doxorubicin 75 mg/m(2) every 2 weeks for three cycles followed by docetaxel 100 mg/m(2) every 2 weeks for three cycles. Patients in arm B received combination therapy with doxorubicin 56 mg/m(2) plus docetaxel 75 mg/m(2) every 3 weeks for four cycles. Granulocyte colony-stimulating factor was administered on days 2 to 12 of each cycle in both groups. RESULTS: Forty patients were entered onto the trial. Pretreatment tumor size averaged 5.7 cm with clinically positive axillary lymph nodes in 23 patients (57%). As expected, myelosuppression was severe in both groups; however, >/= 80% of planned dose-intensity was delivered. Hand-foot syndrome was more common after sequential therapy. Clinical responses were similar in both groups, with an overall response rate of 87%, including 20% clinical complete remissions. Pathologic complete remission or residual in situ disease only was confirmed in five patients (12.8%). Patients who received sequential therapy had fewer positive lymph nodes (mean, 2.17 v 4.81; P <.037) at definitive surgery. CONCLUSION: Primary chemotherapy with doxorubicin and docetaxel is well tolerated and highly active. A sequential treatment schedule increases toxicity but may result in more substantial lymph node clearance than combination therapy.     

A simple index using video image analysis to predict disease outcome in primary breast cancer.

Image analysis was used to investigate the prognostic significance of immunostaining for oestrogen receptor (ER), p53 tumour-suppressor protein and tumour cell proliferation (MIB-1) in a random cohort of 200 primary breast cancer patients with between 4 and 7 years of clinical follow-up. Image measurements of the percentage of immunopositive cancer cell nuclei (% positive nuclear area) were recorded for the above tumour features for each patient. Assessment of relative risk using Cox's univariate analysis indicated that tumour size, number of cancer-involved nodes, MIB-1 and ER % positive nuclear area were significantly associated with breast cancer disease outcome, i.e., relapse-free survival and overall survival. In multivariate analysis, tumour size, number of involved nodes, ER and MIB-1 % positive nuclear area were retained as independent predictors of prognosis, depending on the image measurement cut-point used. A prognostic model, which can be used without reference to nodal involvement, was constructed for tumour size, ER cut-point of 30% positive nuclear area and MIB-1 cut-point of 10% positive nuclear area. Kaplan-Meier analysis of this image-based prognostic index identified 4 risk groups with predicted 5-year overall survival rates of 93%, 83%, 76.7% and 61.5%. We conclude that image measurements of ER and proliferative rate can be combined with tumour size to construct a prognostic index which reliably predicts disease outcome in primary breast cancer without knowledge of the nodal status of the patient.