Age-related DNA methylation changes in normal human prostate tissues.

PURPOSE: Prostate cancer is a leading cause of cancer death among the aging male population but the mechanism underlying this association is unclear. Aberrant methylation of promoter CpG islands is associated with silencing of genes and age-dependent methylation of several genes has been proposed as a risk factor for sporadic cancer. We examined the extent of gene methylation in pathologically normal human prostate as a function of age. EXPERIMENTAL DESIGN: We used pyrosequencing to quantitatively analyze the methylation status of nine CpG islands in normal prostate tissue DNA from 45 organ donors and 45 patients who had undergone cystoprostatectomy for bladder cancer. We also analyzed 12 pairs of matched benign and prostate cancer tissue DNA from patients with prostate cancer. RESULTS: Linear regression analysis revealed a significant increase in promoter methylation levels correlating with age for CpG islands at RARbeta2 (r = 0.4; P < 0.0001), RASSF1A (r = 0.27; P = 0.01), GSTP1 (r = 0.59; P < 0.0001), NKX2-5 (r = 0.27; P = 0.008), and ESR1 (r = 0.244; P = 0.023) in the normal prostate tissue samples studied. A calculated average methylation (z score) at all nine CpG loci analyzed in the normal prostate tissues showed a strong correlation with age (r = 0.6; P < 0.001). Comparison of the methylation level for the matched benign and prostate cancer tissues from individual patients with prostate cancer showed significantly higher methylation in the prostate cancer tissue samples for RARbeta2 (P < 0.001), RASSF1A (P = 0.005), GSTP1 (P < 0.001), NKX2-5 (P = 0.003), ESR1 (P = 0.016), and CLSTN1 (P = 0.01). CONCLUSIONS: Our findings show aberrant hypermethylation as a function of age in the normal prostate tissues. Such age-related methylation may precede and predispose to full-blown malignancy.     

A study of the conditions of the supercritical fluid extraction in the analysis of selected anti-inflammatory drugs in plasma

Supercritical fluid extraction (SFE) was employed to analyze selected anti-inflammatory drugs in plasma. Evaluation of selected drugs (ibuprofen, indomethacin, and flufenamic acid) was performed using the HPLC method on columns with the reverse phase C-18 and detection in the UV region of the spectrum. A study of the conditions of SFE carried out for 30 min at 50 degrees C investigated the magnitude of the pressure of carbon dioxide suitable for drug extraction, the selection of the collecting solvent, and the modification of CO2 with an organic solvent. The results of the study made it possible to determine the optimal procedure for SFE of ibuprofen, indomethacin, and flufenamic acid from plasma, which renders their HPLC quantification possible.     

Renal ischemia-reperfusion injury in the rat is prevented by a novel immune modulation therapy

BACKGROUND: Vasogen Inc.'s (Mississauga, Ontario, Canada) immune modulation therapy (IMT) is a therapy in which cells from the patient's own blood are modified by ex vivo exposure to specific physicochemical stressors, including oxidation, ultraviolet (UV) light, and an elevated temperature. The therapy has been shown to have a beneficial effect in models of inflammation and vascular diseases. This study tested the hypothesis that IMT can prevent renal ischemia-reperfusion (I/R) injury in rats. METHODS: Whole blood was collected from syngeneic age-matched donors by cardiac puncture. It was treated with a combination of controlled physiochemical stressors consisting of elevated temperature, a gas mixture of medical oxygen containing ozone, and UV light. The treated blood (150 microL) was injected in the gluteal muscle. Control animals received the same volume of untreated blood or physiological saline. Transient (45 or 60 minutes) left-renal ischemia was produced with simultaneous contralateral nephrectomy in treated and control spontaneously hypertensive rats (SHR). Young and old male and female rats were studied. Plasma creatinine, diuresis, and the survival rates of each group were compared. Renal apoptosis-necrosis was estimated by DNA laddering, histology, and in situ terminal deoxynucleotidyl transferase assay. mRNA levels of several regulators of apoptosis-regeneration were determined in control and postischemic kidneys by Northern blotting. RESULTS: IMT pretreatment of SHR significantly reduced renal I/R injury compared with equivalent placebo treatments consisting of untreated blood- or saline-injected SHR, as evidenced by a significant increase of the survival rate curves in young and old male SHR, which correlated with 24-hour postischemic diuresis. The increases in plasma creatinine following renal I/R were significantly lower in IMT-treated young male and old female SHR compared with saline or untreated blood-injected controls. Dilution analysis showed that the protective effect of treated blood was lost by dilution. Loss of epithelial cells was reduced in IMT-treated rats, with a significant decline in the peak of apoptosis 12 hours after acute ischemic renal injury. IMT did not modify the pattern of mRNA levels of several genes involved in the inflammation and regeneration processes. CONCLUSION: Our data demonstrate that IMT prevents the destruction of kidney tissue and the resulting animal death caused by renal I/R injury.     

Microsatellite instability in hematological malignancies

The replication error (RER+) phenotype, characterized by microsatellite instability (MSI) has been recently related to mutations of genes involved in DNA mismatch repair pathway. These genetic alterations were first described in hereditary non polyposis colorectal cancer (HNPCC). We examined 44 patients with hematological malignancies (27 AML, 9 MDS, 2 CML-BP and 6 T-ALL) for evidence of MSI. Twenty seven percent of our patients showed differences for only one marker. In four cases (9.1%) MSI was observed in multiple markers and these cases were described as RER+ phenotype. Presented data suggest that this phenomenon may play a role in at least a subset of patients with hematological malignancies.     

Evaluation of geometric and dosimetric inaccuracies of stereotactic irradiation in the rat brain

OBJECTIVE: To evaluate geometric and dosimetric inaccuracies in the irradiation of the rat brain with the Leksell Gamma Knife. MATERIALS AND METHODS: Altogether three types of dosimeters were employed for these measurements: (a) a thermoluminescent dosimeter, (b) a semiconductor detector and (c) a polymer gel dosimeter. The thermoluminescent dosimeter and the semiconductor detector were calibrated using an ion chamber and then implanted in the brain of a rat cadaver and used for absolute dose determination. A special glass phantom mimicking exactly the shape of the rat body filled with the polymer gel was used for measurements of the relative dose distribution and evaluation of geometric inaccuracies during the stereotactic irradiation in the rat brain. RESULTS: Both thermoluminescent and semiconductor detectors, due to their size, measured mean doses. The observed results demonstrated that the Leksell GammaPlan can be employed for the calculation of absorbed doses in irradiation of experimental animals. In our case, it was necessary to apply a correction factor of 1.078 for the absolute absorbed dose to obtain reliable results. A comparison of calculated dose profiles using the treatment planning system in all three axes with those measured by the polymer gel dosimeter demonstrated a very good geometric agreement with the mean deviation in profile position of 0.5 mm. CONCLUSION: The results indicate that this technique can effectively check the geometric and dosimetric accuracy of stereotactic irradiation in the rat brain. The Leksell GammaPlan can be employed for the calculation of absorbed doses, but the correction factor of 1.078 had to be applied for the absolute dose calculations in our irradiation geometry.     

Über Versuche zur Normalisierung einer gestörten Darmflora beim Säugling

Ohne Zusammenfassung     

Polymerization of lactams, 58. Nonactivated polymerization of 2-pyrrolidone in the presence of higher lactams

Addition of a higher lactam (ε-caprolactam or 8-octanelactam) to the nonactivated polymerization of 2-pyrrolidone, initiated with potassium tert-butoxide or the isolated potassium salt of 2-pyrrolidone, does not lead to the formation of a copolyamide. Only a marked decrease of polymerization rate is observed and the maximum rate of 2-pyrrolidone homopolymerization shifts to a lower temperature. The possible causes of this interesting phenomenon are discussed.     

Internal mobility of PMMA in concentrated CDCl3 solutions, 2. 13C NMR spectra

The ¹³C NMR relaxation parameters (T₁, line widths and NOE factors) were measured and analyzed for two samples of atactic poly(methyl methacrylate) (PMMA) of molecular weight M_w 4,20·10⁴ and 1,76.10⁶ in CDCl₃ solutions in the concentration range 11 to 54% (by weight) of polymer, at 297 and 323 K. By this analysis it was found that the backbone segmental motion of PMMA is spatially isotropic in the whole measured concentration range; the motion of the α-CH₃ group may be approximated by the double reorientation model; the motion of the ester CH₃ group can be approximated by the isotropic model only at concentrations lower than 12%; in more concentrated solutions, a satisfactory approximation is obtained neither by the isotropic, nor by the double reorientation models. The reasons of this behaviour are discussed.     

Search for XYY syndrome in psychiatrically disturbed children and adolescent juvenile delinquents

Two groups of young males were analyzed: 18 patients from the Division of Child Psychiatry, University of Minnesota, and 28 juvenile delinquents from Lino Lakes Juvenile Detention Center. All patients were screened regarding stature and behavior characteristics. None of the 46 cases analyzed showed any abnormality, either of autosomal or of sex chromosomes.Presented at the Fifth World Congress of Psychiatry, November 25–December 4, 1971, Mexico.     

On the copolymerization of acrylonitrile with styrene in the presence of Lewis acid, 2. Regulation of molecular weight

The results of the study of the acrylonitrile (AN)-sytrene (St) copolymerization in the presence of zinc chloride and various hydrogen donors (cyclohexane, octane, isooctane, and decahydronaphthalene) are reported. It was found that the rate of copolymerization (R_p) increases with concentration of the hydrogen donor at constant concentrations of comonomers in acetone in the presence as well as in the absence of a free-radical source (AIBN). The dependence of the reciprocal value of limiting viscosity number (1/[η]) of the equimolar copolymers on R_p in the system St? AN? ZnCl₂? AIBN is not linear. The position of the minimum on this curve at a given ratio of comonomers depends on the concentration of both zinc chloride and AIBN. The reduction of [η] of the equimolar copolymer attained by the effect of hydrocarbon is relatively small provided the reaction system is homogeneous. On the basis of the results obtained, it may be stated that a change in molecular weight of the copolymer is to be achieved more easily by a change in the rate of copolymerization (effected by AIBN) than by the effect of hydrogen donor (chain transfer).