Determination of in vitro activity renin inhibitors by HPLC method

A high-performance liquid chromatographic assay has been developed to the separation of angiotensin I, tetradecapeptide and the tetrapeptide Leu-Val-Tyr-Ser. This purpose is achieved in a single step, using HPLC technique in the reversed phase system. The method is based on enzymatic hydrolysis of a substrate renin (TDP) with formation of angiotensin I (DP). After 1 h incubation at 37 degrees C the reaction mixture is introduced directly into the chromatographic system. A 200 microl reaction mixture is needed for analysis. Acetonitrile gradient in 0.01 M ammonium acetate buffer allows a satisfactory separation of hydrolysis products. The application of this technique for the determination of in vitro renin activity and evaluation the potency of human renin inhibitors, employing the artificial renin substrate tetradecapeptide, is demonstrated.     

Multiple cerebral glioma or tumor dissemination via CSF pathways? Case report

A case is presented of a 38-year-old female patient who developed bifocal metachronous cerebral glioma with the same histological appearance (glioblastoma multiforme). Two separate tumors were operated on within six months: the first one was localized in the left parieto-occipital area, and the other in the right temporal lobe. The tumor cells dissemination occurred probably via the CSF pathways: during the first operation the posterior horn of the left lateral ventricle was opened, and the second neoplastic lesion was situated also in the direct vicinity of CSF spaces (the Sylvian cistern). For all practical purposes, the case presented testifies to the necessity of intraoperative protection of the CSF spaces by separating any open CSF cisterns from the removed tumor mass with cotton pads. In case of diagnosing cerebral glioma, a possibility of the presence of multiple foci should be taken into account. It is especially important for the differentiation of multiple lesions occurring synchronically, where similarity of radiological features is seen in metastases, cerebral abscesses and demyelinating lesions.     

Suppression of human brain tumor with interference RNA specific for tenascin-C

Glioblastoma multiforme (GBM) accounts for approximately 12-15% of intracranial neoplasms. The GBM remains refractory to therapy because of tumor heterogeneity, local invasion, and non-uniform vascular permeability to drugs. Patients with GBM have the median survival of approximately 8-10 months, and for those cases where tumor recurs, the average time of tumor progression after therapy is only eight weeks. A combination of different treatment modes as surgery and chemo- or/and radiotherapy extend survival only for a short time, if any. Recently, tenascin-C (TN-C) as a dominant epitope in glioblastoma has been discovered. It is transiently expressed during organogenesis, absent or much reduced in most fully developed organs, but reappears under pathological conditions such as infection, inflammation, or tumorigenesis. It was found that the intensity of TN-C staining correlates with the tumor grade and that the strongest staining indicates poor prognosis.     

Fluvastatin does not elevate periosteal osteogenesis induce by Moloney sarcoma virus (MSV) in mice

Several studies have demonstrated the pleiotropic effects of statins. Since Wang and associates reported that in rabbits lovastatin reduced steroid-induced bone loss, numerous authors have confirmed these data, however, others have reported conflicting results. In this study, the effects of fluvastatin on bone formation were investigated in early and late phase of osteogenesis. In the first set of experiments (early phase of osteogenesis) CFW/Ll mice were randomly divided into three groups. Two groups were injected with Moloney-murine sarcoma virus (Mo-MSV) into right thighs to induce orthotopic bone formation. Mice in the experimental group received fluvastatin for 11 consecutive days. Thirty days after Mo-MSV inoculation, total serum cholesterol, triglycerides, high- and low-density lipoprotein cholesterol, alkaline phosphatase (AP) were measured and bone mineral increase was calculated. In the second set of experiments (late phase of osteogenesis), fluvastatin was administered from day 11 after Mo-MSV inoculation for 20 consecutive days. Fluvastatin administration in the early phase of osteogenesis made no significant difference in average bone increase compared with mice receiving placebo. Lipid profile and AP were not significantly affected. During late phase of osteogenesis, the average increase in femural dry mass was significantly lower in the group of mice receiving fluvastatin than in the control group. Also, Mo-MSV-initiated tumors disappeared earlier in mice treated with fluvastatin. This may be attributed to the antioncogenic potential of fluvastatin. These results also point out that orthotopic bone formation at the sites of Mo-MSV inoculation in mice seems to be a useful model to examine the pleiotropic effects of statins.     

Induction of caspase 3 and modulation of some apoptotic genes in human acute promyelocytic leukemia HL-60 cells by carboplatin with amifostine

The influence of carboplatin alone and carboplatin in combination with cytoprotective agent amifostine on the growth, caspase 3 activity and some apoptotic genes expression was investigated in vitro in human acute promyelocytic leukemia HL-60 cells. Proliferation of HL-60 cells exposed to carboplatin dropped down with increasing dose of the drug. This effect was slightly higher when carboplatin was used in combination with amifostine. The cytotoxic index (IC50) was estimated as 6.6 and 4.4 x 10(-4) M (after 24 h) and 3.3 and 2.5 x 10(-5) M (after 48 h) for carboplatin and carboplatin with amifostine, respectively. This effect was accompanied by induction of caspase 3 activity. HL-60 cells treated with carboplatin alone showed about 120-fold increase in caspase 3 activity. Combination of carboplatin with amifostine induced the enzyme activity up to 280 times. Furthermore, the expression of bcl-2, c-myc and bax genes involved in apoptosis as well as p65, which function in this process is unknown, were determined. Semi-quantitative RT-PCR showed a decrease in bcl-2 and an increase in bax, c-myc and p65 expression in HL-60 cells treated with carboplatin in combination with amifostine as compared to the cells treated only with carboplatin. We conclude that amifostine may potentiate carboplatin therapeutic efficiency towards human acute promyelocytic leukemia cells.     

Maternal serum and amniotic fluid IL-1 alpha, IL-1 beta, IL-6 and IL-8 levels in preterm and term labor complicated by PROM

Cytokines may be implicated in the pathophysiologic mechanisms of preterm and term labor. Many studies indicate cytokines as predictors of preterm delivery and explain partially mechanism of preterm uterine contractions. Complicated relations between mediators in systemic fluids of a fetomaternal unit require further explorations. The right diagnosis and management require better understanding of these relationships. OBJECTIVES: The comparison of IL-1 alpha, IL-1 beta, IL-6 and IL-8 levels in maternal serum and amniotic fluid in term and preterm labor complicated by PROM. MATERIAL AND METHODS: In 44 patients in premature labor with PROM (group I) and 33 patients in labor at term with PROM (group II) cytokines levels were estimated one time in amniotic fluid: just after PROM, and two times in maternal serum: just after PROM and during labor. RESULTS: Amniotic fluid cytokines levels were significantly higher in group I than in group II. Maternal serum cytokines concentrations of IL-1 alpha and IL-1 beta in group I were significantly higher than in group II. IL-6 level was significantly higher in group II than in group I. In both groups maternal serum IL-6 levels during labor significantly increased in comparison to IL-6 levels just after PROM. No correlations between amniotic fluid and maternal serum cytokine levels at PROM were observed. CONCLUSIONS: Higher amniotic fluid cytokines levels in patients with preterm labor complicated by PROM than in labor at term with PROM indicate possible differences between PROM mechanisms in preterm and term labor. The increase of IL-6 level during labor can be related with the possible role of this cytokine in the immunological mechanism of the labor beginning. No relationships between amniotic fluid and maternal serum levels of investigated cytokines in PROM suggest the presence of the barrier stopped cytokines transfer by the placenta and the complete separation of these two compartments.     

Evaluation of breast conserving therapy (BCT) standards for invasive and preinvasive breast cancer adapted in RCO in Bydgoszcz

INTRODUCTION: Prophylactic campaign against breast cancer wide spread by Regional Center of Oncology made significant influx of patients coming to our place and presenting less advanced stages of the mentioned disease. AIM: The aim of this work was estimate of BCT treatment standard for invasivum cancer and preinvasivunm breast cancer treatment standard which was adapted in RCO. MATERIALS AND METHOD: Patients presenting unifocal invasive cancer smaller than 2 cm, stage No, where the minimal margin of more than 1 cm was possible to perform, were qualified to BCT. Minimal surgical border during tumorectomy was 1 cm. We were excluded patients with carcinoma lobulare and carcinoma mucinosum. During qualification to DCIS treatment standard first we must exclusion multifocalis cases based on Anderson classification and Falun consens. In the other cases we based on Van Nuys Prognostic Index (VNPI) considering the patient's age. We have 3 forms of treatment: simplex mastectomy, tumorectomy and tumorectomy with RTG-therapy. To BCT standard were qualified 52 patients. Schematically was attended 45 peoples. We had 5 patients which was attended based on preinvasivum breast cancer treatment standard. RESULTS: The size of breast cancer tumor at the patients which was attended based on BCT standard was 0,5-2 cm. 3 patients was disqualified from BCT because we found second breast cancer focus in histopathological material. We must widen surgical border post tumorectomy in 5 cases. Among 5 patients with preinvasivum cancer was 3 simplex mastectomy and 2 cases tumorectomy with RTG-therapy. CONCLUSIONS: In our opinion BCT treatment standard and breast preinvasivum cancer treatment standard should be using only in high specialty oncological center. It is a guaranty of right qualification and treatment for breast cancer patients.     

Preparation of Carbamoyl Derivatives of Indole and of Derivatives of 1,2,4-Triazol-3-one from Semicarbazides

A simple and one-step method for the preparation of the title compounds is described. Semicarbazides 2 react easily with levulinic acid in the presence of HCl to give 4 , whereas in a alkaline medium they cyclize to form the triazole derivatives 3 .