Stage-related increase in the proportion of apoptotic germ cells and altered frequencies of stages in the spermatogenic cycle following gestational, lactational, and direct exposure of male rats to p-nonylphenol.

The cumulative effects of environmental toxicants, for example, the alkylphenol, para-nonylphenol (p-NP) are of concern. Our previous study showed that p-NP reduced several testicular morphometric parameters, including sperm counts. The present study reexamined material collected in that study to determine the mechanistic basis of p-NP action on spermatogenic development in the offspring. Seven-day pregnant Sprague-Dawley rats were treated with vehicle or 100 or 250 mg/kg p-NP through gestation, lactation and afterward directly to all male offspring until 10 weeks of age. Both doses of p-NP significantly (P < 0.02) increased the number of germ cells with in situ end-labeled fragmented DNA (TUNEL positive) by 1.9-fold and 1.7-fold, respectively, and specifically in stages XII-XIV and I-III. TUNEL-labeling was, however, selective, and excluded labeling of basal cells with apoptotic morphology. Cleaved caspase-3 immunohistochemistry strongly labeled basal cells (spermatogonia and early spermatocytes) with condensed marginated chromatin but not degenerate germ cells lacking definitive nuclear material found throughout the epithelium. Only the caspase index (ratio of number of caspase positive to number of degenerate cells) of the 100-mg/kg p-NP group was significantly (p < 0.05) threefold greater than controls. Whereas both doses and either 250 or 100 mg/kg treatment alone significantly (p < 0.002) reduced the frequencies (duration) of stages I-III, VII-VIII, and late VIII-IX (spermiating and recently spermiated tubules), respectively, both doses significantly (p < 0.002) increased the frequencies of stages IV-VI and all stages containing late-stage spermatocytes (XII-XIII) and meiotic cell divisions (XIV). Thus, p-NP, an environmentally persistent xenoestrogen, insidiously alters the spermatogenic cycle and spermatogenic process in male offspring.     

The value of different resistance parameters in distinguishing biopsy-proved dysfunction of renal allografts

The data concerning the value of duplex sonography in diagnosingparenchymatous renal allograft dysfunction are controversial.Most early studies did not take into consideration the manyfactors influencing resistance parameters. We therefore performeda prospective, biopsy-controlled study with exclusion of allknown sources of error regarding resistance parameters. Furthermorewe investigated the value of a new resistance parameter, thesystolic deceleration percentage. Forty-seven duplex sonographicstudies were performed on 43 patients (30 male, 13 female, medianage 47 years, range 7–70). Fourteen studies were doneon normally functioning grafts (control group) an average of33 days after transplantation. Thirty-three studies were performedon dysfunctional grafts immediately prior to biopsy. Graftswhich had been transplanted more than a year previously or withvascular findings or any other clinical or sonographic pathologyprobably explaining function deterioration were excluded. Inall patients, the resistive index (RI), pulsatility index (PI)and systolic deceleration percentage (DP) were calculated inthe main renal artery and in the interlobar artery. Of the 33grafts with dysfunction, nine had vascular rejection (VR), 11interstitial rejection (IR), 11 cyclosporin A toxicity (CAT)and two other histologies (OR). The mean RI in normal grafts(NO) was 0.71±0.06 in the main artery and 0.68±0.06in the interlobar artery, in VR 0.86±0.12 and 0.80±0.18,in IR 0.72±0.05 and 0.70±0.07, in CAT 0.67±0.06and 0.65±0.07 and in OR 0.64±0.07 and 0.60±0.01.For PI, the values were 1.45±0.23 and 1.41±0.28(NO), 3.5±2.13 and 2.92±2.16 (VR), 1.55±0.26and 1.46±0.33 (IR), 1.32±0.25 and 1.27±0.26(CAT) and 1.30±0.34 and 1.13±0.04 (OR). For DPwe calculated 28±5% and 29±6% (NO), 43±14%and 36±6% (VR), 29±9% and 27±9% (IR), 31±8%and 32±7% (CAT ) and 32±4% and 28±3% (OR).The sensitivity/specificity for VR with a cutoff mean+2 SD was0.44/1 for RI, 0.55/0.97 for PI and 0.33/0.89 for DP. It wasconcluded that:(1) despite the high selection of our patientgroup, diagnostic accuracy of duplex sonography for diagnosingparenchymatous function disorder in renal allograft remainsinsufficient; (2) in vascular rejection only, the resistanceparameters differ significantly from the values of normal allografts;(3) the higher the cutoff of resistance parameters, the betterthe specificity and the worse the sensitivity for diagnosingvascular rejection; (4) of all investigated resistance parameters,the RI is the most practical due to a simple measurement technique.     

PET for evaluation of differential myocardial perfusion dynamics after VEGF gene therapy and laser therapy in end-stage coronary artery disease.

The purpose of this study was to appraise the value of PET in the assessment of the effect of supposedly proangiogenic new therapies such as gene therapy with vascular endothelial growth factor (VEGF) gene and endomyocardial laser therapy. METHODS: Thirty-five patients with end-stage coronary artery disease and class III (Canadian Cardiovascular Society) angina were included. Myocardial ischemia was evaluated with dipyridamole PET scanning and exercise tolerance with bicycle ergometry. Ten patients were treated with naked plasmid DNA encoding for human VEGF165 (VEGF) and 12 patients were treated with laser therapy (direct myocardial revascularization [DMR]) using an electromechanical mapping system. Thirteen patients were treated with standard medical therapy (control). RESULTS: In both active treatment groups, angina was reduced in most subjects, except in 2 VEGF and 5 DMR patients. In the control group, no improvement in anginal classification was found, except in 3 subjects. On the PET scan, solely in the VEGF group, the stress perfusion was significantly improved (from 57 +/- 33 to 81 +/- 55 mL/min/100 g; P = 0.031). Furthermore, in the VEGF group, the number of ischemic segments was reduced from 274 +/- 41 to 234 +/- 48 segments (P = 0.004) but not in the DMR group (from 209 +/- 43 to 215 +/- 52 segments) or in the control group (from 218 +/- 18 to 213 +/- 28 segments). Bicycle exercise duration showed slight nonsignificant changes in the VEGF group (from 3.6 +/- 2.0 to 4.6 +/- 2.1 min), in the DMR group (from 5.1 +/- 1.5 to 4.7 +/- 1.3 min), and in the control group (from 3.3 +/- 1.8 to 3.5 +/- 2.3 min). CONCLUSION: PET showed that intramyocardial gene therapy with the human VEGF165 gene in contrast to laser DMR treatment effectively reduces myocardial ischemia.     

Die Lungencirculation und der arterielle Blutdruck

Ohne ZusammenfassungVorliegende Arbeit wurde schon vorgenommen im Winter 1880–1881, als ich noch Assistent am physiologischen Institut der Universität Leiden war. Durch besondere Umstände konnte sie erst heute publicirt werden.     

The endoscopic spectrum of primary non-Hodgkin's lymphoma of the stomach

Thirty-one consecutive patients with primary non-Hodgkin's lymphoma of the stomach were studied to outline the spectrum of endoscopic abnormalities. The 17 men and 14 women had a median age of 65 years. There were 22 patients in stage I and 9 in stage II. Three endoscopic patterns were recognized: diffuse infiltration (9), ulceration (9) and polypoid lesions (13). There was no apparent correlation between the endoscopic appearance and the grading or subtype of the malignancy. Local recurrence did not occur once complete remission was obtained. The disease-free 5-year survival rate was 45%. Recognition of these endoscopic patterns may lead to earlier detection and, hopefully, improved survival.     

Tumor vascular signals in renal masses: detection with Doppler US

The vascularity of 49 renal masses (26 malignant and 23 benign lesions) was investigated with duplex Doppler ultrasound. Doppler signals obtained at the margins of renal masses were defined as "tumor signals" when the Doppler-shifted frequency of the lesion exceeded the frequency shift in the ipsilateral main renal artery. These exceeded 2.5 kHz with a 3-MHz insonating frequency. Among the 26 renal masses that subsequently proved to be malignant, tumor signals were obtained in 15 of 18 (83%) untreated renal cell carcinomas, in three of four Wilms tumors, and in two patients with metastases to the kidney, but not in the one patient with lymphoma. None of the 23 benign renal masses demonstrated tumor signals. Tumor vascularity in malignant lesions gives rise to abnormal, high-velocity, Doppler-shifted signals that can help in the differential diagnosis of renal masses.     

Simultaneous Detection of 15 Human Cytokines in a Single Sample of Stimulated Peripheral Blood Mononuclear Cells

Cytokines secreted by cells of the immune system can alter the behavior and properties of immune or other cells. At a site of inflammation, sets of cytokines interact with immune cells, and their combined effect is often more important than the function of one isolated component. Conventional techniques, such as enzyme-linked immunosorbent assays, generally require large quantities of cells to characterize a complete cytokine profile of activated lymphocytes. The Bio-Plex system from Bio-Rad Laboratories combines the principle of a sandwich immunoassay with the Luminex fluorescent-bead-based technology. We developed a multiplex cytokine assay to detect different cytokines simultaneously in culture supernatant of human peripheral blood mononuclear cells stimulated with antigen and with mitogen. Fifteen human cytokines (interleukin 1α [IL-1α], IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-15, IL-17, IL-18, gamma interferon, and tumor necrosis factor alpha) were validated with a panel of healthy individuals, rheumatoid arthritis patients, and juvenile idiopathic arthritis patients. Comparing the multiplex assay with a regular enzyme-linked immunosorbent assay technique with this donor panel resulted in correlation coefficients for all cytokines ranging from 0.75 to 0.99. Intra-assay variance proved to be less then 10%, whereas interassay variability ranged between 10 and 22%. This multiplex system proved to be a powerful tool in the quantitation of cytokines. It will provide a more complete picture in differences between activated lymphocyte cytokine profiles from healthy individuals and those from patients with chronic inflammatory diseases.