Urinary Lipidomics: evidence for multiple sources and sexual dimorphism in healthy individuals.

Urinary lipidomics may add new valuable biomarkers to the diagnostic armamentarium for early detection of metabolic and kidney diseases. Sources and composition of urinary lipids in healthy individuals, however, have not been investigated in detail. Shotgun lipidomics was used to quantify lipidomic profiles in native urine samples from 16 individuals (eight men, eight women) collected in five fractions over 24 h. All probands were comprehensively characterized by urinary and clinical indices. The mean total urinary lipid concentration per sample was 0.84 µM in men and 1.03 µM in women. We observed significant intra- and interindividual variations of lipid concentrations over time, but failed to detect a clear circadian pattern. Based on quantity and subclass composition it seems very unlikely that plasma serves as major source for the urinary lipidome. Considering lipid metabolites occurring in at least 20% of all samples 38 lipid species from 7 lipid classes were identified. Four phosphatidylserine and one phosphatidylethanolamine ether species (PE-O 36:5) were detectable in almost all urine samples. Sexual dimorphism has been found mainly for phosphatidylcholines and phosphatidylethanolamines. In men and in women urinary lipid species were highly correlated with urinary creatinine and albumin excretion, reflecting glomerular filtration and tubular transport processes. In women, however, lipid species deriving from urinary cells and cellular constituents of the lower genitourinary tract considerably contributed to the urinary lipidome. In conclusion, our study revealed the potential of urinary lipidomics but also the complexity of methodological challenges which have to be overcome for its implementation as a routine diagnostic tool for renal, urological and metabolic diseases.

Age-associated changes in the densities of presynaptic monoamine transporters in different regions of the rat brain from early juvenile life to late adulthood.

The binding parameters of highly selective ligands of serotonin (5-HT) transporters ([3H]paroxetine), noradrenaline (NE) transporters ([3H]nisoxetine), and of dopamine (DA) transporters ([3H]GBR-12935) were determined on membrane preparations from frontal cortex, striatum, midbrain and brain stem of Wistar rats on postnatal days 25, 50, 90 and 240, i.e., from the time of weaning till late adulthood. No age-dependent alterations in the affinity-parameters (K(D)-values) of all three monoamine transporters were observed. Age-associated changes in B(max)-values of the binding of all three specific ligands were most pronounced in the phylogenetically younger, late maturing brain regions (frontal cortex, striatum). Most likely, these changes reflect age-related changes in 5-HT, NE and DA-innervation densities. In the frontal cortex, 5-HT-transporter density increased steadily from weaning (day 25) till late adulthood, whereas the density of NE-transporters was highest at weaning, declined till puberty (day 50) and remained at this level until old age. DA-transporter density in the frontal cortex was not reliably measurable by [3H]GBR-binding assays. In the striatum, DA-transporter density increased till puberty and declined thereafter considerably and steadily to about one-fourth of the pubertal values at old age. No such age-associated changes in DA-transporter density were seen in the midbrain. Densities of 5-HT and NE remained at the level reached already at weaning until old age in the striatum, midbrain and brain stem. These findings provide the first comprehensive description of the normally occurring changes in the densities of all three presynaptically located monoamine transporters in the rat brain throughout the life span from weaning to late adulthood.

Evaluation of tolerance limits for humans under heat stress and the problems involved.

Investigators are divided about the limits of tolerable ambient temperatures and air humidities, even under equal boundary conditions. In 266 climatic chamber experiments, mostly lasting 4 h, 13 young healthy men were exposed to combinations of ambient temperature (20 to 55 degrees C) and relative humidity (5 to 97%) at an air velocity of 0.3 m.s-1. They were lightly clad and performed equal treadmill work (metabolic rate about 850 kJ.h-1). Mean rectal isotherms and their standard deviations were calculated from the final rectal temperatures of the 13 men and plotted on a psychrometric chart. The slopes of the isotherms were compared with those of various "indices of heat stress." The probability of reaching the potentially critical rectal temperature of 39 degrees C was calculated according to data of Wyndham & Heyns. It increased from about 1:500,000 to 1:150 in climates corresponding to mean isotherms of 37.6 and 38.2 degrees C, respectively. Most of the limits thus far proposed are situated between these two isotherms, and they differ merely in regard to health risk. It is useless to discuss "correct" or "incorrect" limits until society has decided what risk is to be accepted. An upper tolerable limit per se does not and cannot exist. For several reasons the 37.6 degrees C isotherm might be taken into consideration as a limit.

Light retardance by human oocyte spindle is positively related to pronuclear score after ICSI.

Disturbed spindle assembly increases risks of chromosome mal-segregation. Non-invasive polarization microscopy (PolScope) was employed in two centres to assess spindle integrity for the first time quantitatively in human oocytes from consenting patients undergoing intracytoplasmic sperm injection (ICSI) with respect to pronuclear (PN) score after fertilization. In one centre oocytes were selected before ICSI, in another selection was after ICSI according to PN score. In both centres, mean retardance of light by birefringent spindles in oocytes forming a pre-embryo with good PN score after ICSI was significantly higher compared with spindles in oocytes developing into a lower PN score pre-embryo with limited developmental potential (P < 0.001). Transfers involving oocytes with high retardance and at least one good PN score embryo resulted more frequently in a conception than transfers from oocytes with spindles of lower mean retardance and lower PN score embryos. There was a trend for an inverse relationship between age and magnitude of retardance in a small oocyte cohort. The study suggests that quantitative evaluation of mean retardance of light by the oocyte spindle predicts oocyte health, is related to PN score of the embryo and may be especially useful to assess oocyte quality in countries with legal restrictions to select after fertilization.

High magnitude of light retardation by the zona pellucida is associated with conception cycles.

BACKGROUND: Failures in expression of zona proteins correlate to subfertility in animals. Low expression of the zona proteins by the growing human oocyte may indicate reduced developmental potential. Therefore, we non-invasively analysed the thickness and the structure of the zona pellucida (ZP) of human oocytes with respect to embryo fate after ICSI. METHODS: Retardance magnitude and thickness of the inner, middle and outer layers of the ZP were quantitatively analysed by a Polscope in 166 oocytes selected for transfer after ICSI (63 patients; 32.8 +/- 4.4 years) on the basis of pronuclear score at day 1. Blastomere number was determined at day 2. Data were compared between conception cycles (CC; 65 oocytes/23 patients) and non-conception cycles (NCC; 101 oocytes/40 patients) and with respect to maternal age. RESULTS: The thickness was slightly elevated (P < 0.001), and the mean magnitude of light retardance was nearly 30% higher (P < 0.001) in the inner layer of the zona pellucida of oocytes contributing to CC compared to NCC. Embryos in the CC group tended to develop faster. CONCLUSIONS: The magnitude of light retardance by the zona pellucida inner layer appears to present a unique non-invasive marker for oocyte developmental potential.

Influence of motility and vitality in intracytoplasmic sperm injection with ejaculated and testicular sperm.

The vitality of spermatozoa used for intracytoplasmic sperm injection (ICSI) is a crucial factor for fertilization, establishment and outcome of a pregnancy in assisted reproductive technique cycles. The sperm origin may also be a limiting factor, although little is known about this issue. It is known that the motility of injected spermatozoa and their origin from ejaculate or testicular biopsies are important predictors in terms of fertilization, pregnancy and birth rates. Oocytes of patients in 2593 cycles were retrieved in our in vitro fertilization programme and inseminated via ICSI. We used motile (group 1, n = 2317) or immotile ejaculated spermatozoa (group 2, n = 79), motile sperm retrieved from testicular biopsies (group 3, n = 62) and immotile spermatozoa from testicular biopsies (group 4, n = 135). Female age and number of oocytes retrieved did not differ significantly among the groups. The fertilization rates were as follows: 67.1% in group 1, 49.8% in group 2, 68.3% in group 3 and 47.8% in group 4. The pregnancy rates in cases where three embryos had been transferred amounted to 35.7% in group 1, 17.3% in group 2, 38.3% in group 3 and 20.5% in group 4. The embryo quality showed no differences between groups 1 and 3 (14.5), and between groups 2 (11.8) and 4 (10.8). The abortion rate was similar in groups 1-3, but increased in group 4 (26.6%, 27.3%, 31.6% and 55.5%). Irrespective of their origin, the fertilization potential of injected spermatozoa was found to be influenced by motility. The resulting pregnancy and birth rates, i.e. the potential of the resulting embryos to implant and to achieve viable pregnancies, seem to be additionally dependent on the sperm origin. This was well shown by declining rates when spermatozoa in a relatively early stage of maturity had been used. We see increasing evidence that the degree of sperm maturity has an important impact on the outcome of ICSI. In obstructive azoospermia, spermatozoa retrieved from the epididymis should be used rather than testicular biopsy spermatozoa, or testicular sperm should be preincubated in culture medium before ICSI.

[The surgical realization of optimal timing for undescended testicle]. / Zur chirurgischen Realisierung des optimalen Therapiezeitpunktes der Hodenfehllagen.

An account is given in this paper of surgical indications to cope with cryptorchism. Views on timing of surgical action have undergone considerable change over the past 35 years, ranging from prepuberty to the second year of age, the latter date being considered optimal now. Optimal timing is not kept in practice, as may be seen from a regional longitudinal study over 25 years as well as from a representative inquiry among 17 centres of paediatric surgery across the GDR over the same period of time and from a cross-section analysis of all boys operated on for cryptorchism throughout the GDR in 1986. All medical doctors serving at infant welfare centres and day nurseries are invited to refer affected boys to surgeons at an earlier date. The existing health system for children and adolescents is considered to provide good conditions for lowering the average age at surgery.

Radiation exposure in endoscopic retrograde cholangiopancreatography.

Radiation exposure was studied on the basis of x-ray screening time and spotfilms taken in 3590 patients undergoing ERCP. Factors influencing radiation exposure in diagnostic procedures were the age of the patient, target duct system and gastroenteric anastomosis, while duodenal diverticula proved to be of no significance. In endoscopic sphincterotomy average fluoroscopic time was about one minute longer. In contrast to diagnostic ERC, there was an appreciable reduction of radiation exposure in endoscopic sphincterotomy with time, reflecting the growing experience of the endoscopic team. The data presented are within the acceptable range for conventional radiological Gl-examinations.

Ribulose bisphosphate carboxylase capacity and chlorophyll content in developing seedlings of Chenopodium rubrum L. growing under light of different qualities and fluence rates.

In order to evaluate the aclimation of Chenopodium seedlings to different quantum fluence rates of R and BL, kinetics of Rubisco capacity, Chl content and chloroplast structure were studied. Under monochromatic light photoreceptors are stimulated selectively and their influence on biosynthetis capacities during chloroplast development can be studied.R irradiations saturate Rubisco capacity even at the lowest quantum fluence rates applied, whereas Chl a+b synthesis depends strongly upon fluence rate of R. Under BL irradiations, both Rubisco capacity and Chl content are fluence rate dependent. R irradiations favour Chl b synthesis relative to Chl a, whereas under BL Chl a content is high relative to Chl b. Under R irradiation Pfr is the main photoreceptor involved in regulation of Rubisco capacity whereas under BL a specific BL absorbing photoreceptor may control the response. From the fluence rate dependency under BL irradiations it is concluded that the blue region of the day light spectrum may be the sensor for monitoring fluence rate and causing the characteristic changes in shade and high/low WL adaptation with respect to Rubisco levels in Chenopodium.