Comparative protein composition of the brains of PACAP-deficient mice using mass spectrometry-based proteomic analysis.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a widespread neuropeptide acting as a neurotransmitter, neuromodulator, or neurotrophic factor. The diverse biological actions provide the background for the variety of deficits observed in mice lacking endogenous PACAP. PACAP-deficient mice display several abnormalities, such as sudden infant death syndrome (SIDS)-like phenotype, decreased cell protection, and increased risk of Parkinson's disease. However, the molecular and proteomic background is still unclear. Therefore, our aim was to investigate the differences in peptide and protein composition in the brains of PACAP-deficient and wild-type mice using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometric (MS)-based proteomic analysis. Brains from PACAP-deficient mice were removed, and different brain areas (cortex, hippocampus, diencephalon, mesencephalon, brainstem, and cerebellum) were separated. Brain pieces were weighed, homogenized, and further processed for electrophoretic analysis. Our results revealed several differences in diencephalon and mesencephalon. The protein bands of interest were cut from the gel, samples were digested with trypsin, and the tryptic peptides were measured by matrix-assisted laser desorption ionization time of flight (MALDI TOF) MS. Results were analyzed by MASCOT Search Engine. Among the altered proteins, several are involved in metabolic processes, energy homeostasis, and structural integrity. ATP-synthase and tubulin beta-2A were expressed more strongly in PACAP-knockout mice. In contrast, the expression of more peptides/proteins markedly decreased in knockout mice, like pyruvate kinase, fructose biphosphate aldolase-A, glutathione S-transferase, peptidyl propyl cis-trans isomerase-A, gamma enolase, and aspartate amino transferase. The altered expression of these enzymes might partially account for the decreased antioxidant and detoxifying capacity of PACAP-deficient mice accompanying the increased vulnerability of these animals. Our results provide novel insight into the altered biochemical processes in mice lacking endogenous PACAP.

Effects of pituitary adenylate cyclase activating polypeptide on human sperm motility.

Pituitary adenylate cyclase activating polypeptide (PACAP), a neuropeptide with diverse effects, was originally isolated as a hypothalamo-hypophyseal peptide. Subsequent studies showed highest levels of PACAP in the testis after the brain, suggesting that it influences the development and functioning of spermatozoa. Indeed, it has been proven that PACAP has an effect on spermatogenesis, both locally and via influencing the hypothalamo-hypophyseal-gonadal axis. The aim of the present study was to determine whether PACAP has an effect on human sperm motility and whether it is present in the human seminal fluid. Furthermore, the sperm head morphology was studied in mice lacking endogenous PACAP. Human samples were obtained from healthy adult volunteers and andrological patients. The effects of PACAP on the motility of human sperm cells were investigated using a computer aided sperm analysis system. In cases where the motility was lower, addition of PACAP to the samples increased the motility and the ratio of rapid progressive and medium progressive sperm motility groups. The presence of PACAP could not be detected in human seminal fluid samples by means of mass spectrometry. Investigating sperm head morphology with routine histology in PACAP deficient mice revealed that both the longitudinal and transverse diameters were significantly lower in PACAP deficient mice, without marked difference in the shape, as revealed by scanning electron microscopy.

Investigation of pituitary adenylate cyclase activating polypeptide in human gynecological and other biological fluids by using MALDI TOF mass spectrometry.

Pituitary adenylate cyclase activating polypeptide (PACAP) is a multifunctional and pleiotropic neuropeptide. PACAP has diverse effects in the endocrine system, among others, it plays important roles in oogenesis, implantation and development of the nervous system. However, it is not known whether PACAP is present in the fluids of the human reproductive organs. The aim of the present study was to determine, by means of mass spectrometry and radioimmunoassay, whether PACAP is present in human amniotic fluid, ovarian follicular fluid and cervico-vaginal fluid. Samples were obtained from healthy adult volunteers. Our MALDI TOF and MALDI TOF/TOF spectrometry results show that PACAP38 is present in all of the follicular fluid samples, and PACAP-like immunoreactivity was also measured by radioimmunoassay. However, we did not find the characteristic peak representing the unmodified 38 amino acid form of the peptide in normal cervico-vaginal smear and amniotic fluid samples. Furthermore, we analyzed other body fluids for comparison, such as human nasal fluid, saliva and aqueous humor. PACAP was not found in these latter samples. In summary, the present study provides evidence for the presence of PACAP in human follicular fluid, suggesting a role in oocyte function, but determination of the exact physiological significance awaits further investigation.

Association among fitness components, anxiety, and confidence following aerobic training in aquarunning.

The relationship of actual and perceived physiological and physical changes, i.e., VO2max, percent body fat, body weight, and perceived fitness change, to changes in anxiety and confidence were investigated. 15 adults volunteered to participate in an 8-wk. exercise program and 15 adults volunteered to be in a nonexercising control group. Physiological and physical measures (VO2max, percent body fat, body weight) were taken at Week 1 (pretest week) and Week 10 (posttest week) of the program. Each subject completed the Competitive Sport Anxiety Inventory-II prior to and just after the 8-wk. training program. Cognitive and somatic anxiety scores significantly decreased for both groups over the 8-wk. period. Confidence scores did not change for either group. The physiological variable and the physical measures were significantly related to cognitive anxiety, somatic anxiety, or confidence. It appears that both the aquarunning exercise and quiet rest sessions can be associated with anxiety within 8 wk.

[Changes in approach to the delivery of dysmature infants following the introduction of Doppler flowmetry]. / A Doppler-flowmetria bevezetését követö szemléletváltozás a dysmaturus magzatok szülésvezetésében.

The authors studied the clinical history dysmature newborns delivered between 1982-1988. Since the middle of 1985 the fetal circulation in each case has routinely been followed by Doppler flowmetry. With this direct assessment of fetal circulation it is possible to diagnose intrauterine growth retardation and pathologic deviations of fetal circulation at earliest. As a result of this the authors have found an increase in the frequency of terminated pregnancies and caesarean sections. The ratio has been shifted towards pre and dysmature newborns. But on the other hand the frequency of intrauterine deaths has decreased. In all those cases where the dysmature newborn died during the perinatal period there were pathologic flow parameters, as well as malformations in some cases. Further prospective clinical studies of neurological development and disorders of dysmature newborns and children are necessary to decide whether our new point of view has been right or wrong.

[Importance of arginine content and arginase activity in fertility]. / Die Rolle des Arginingehaltes und der Arginase-Aktivität in der Fertilität.

Experimental studies have shown that in men with pathospermia the arginine content of sperm plasma is lower than in normal men. In order to find out the reason of this divergency the authors analysed the content of arginine, urea, citrulline, GAA and arginase activity in serum and in sperm plasma of 40 patients with normo-, oligo- or azoospermia. For the first time our results show that the low arginine content in pathological sperm is not due to disorder of the precursor metabolism, but to the increased arginase activity and to the increased conversion of arginine to GAA. In order to find out the mechanism of action and to develop an adequate andrological therapy detailed studies on molecular level have to be done.