Correlation of beta-endorphin and prostaglandin E2 levels in prostatic fluid of patients with chronic prostatitis with diagnosis and treatment response.

PURPOSE: The chronic pelvic pain syndrome is a clinically defined symptom complex of unclear etiology. We have noted increased oxidative stress in the prostatic fluid of these patients, implying an active inflammatory response. Immune cells can produce the natural opioid beta-endorphin at the site of injury, which may modulate pain. We measured beta-endorphin and the inflammatory marker prostaglandin E2 in the expressed prostatic secretions of men with prostatitis, and correlated the results with symptoms. MATERIALS AND METHODS: Expressed prostatic secretions samples from 70 patients and 8 asymptomatic controls were collected and frozen. beta-Endorphin and prostaglandin E2 were measured by enzyme-linked immunosorbent assay. Results were stratified according to prostatitis category and compared in individuals before and after therapy. RESULTS: In symptomatic patients beta-endorphin and prostaglandin E2 were not significantly different in categories II, IIIa and IIIb expressed prostatic secretions but they were higher than in controls. The mean beta-endorphin level plus or minus standard error of mean in symptomatic patients was significantly higher (23.8 +/- 11 ng./ml. versus 8.7 +/- 4.7, p = 0.0001) and mean prostaglandin E2 was lower (6.01 +/- 2.9 ng./ml. versus 3.01 +/- 2.9, p = 0.001) after successful therapy with antibiotics or antioxidant phytotherapy, Prosta-Q (Farr Laboratories, Santa Clarita, California). CONCLUSIONS: We observed a correlation of higher prostaglandin E2 and lower beta-endorphin in symptomatic men with chronic prostatitis. Increased oxidative stress and inflammation may induce prostaglandin E2 production that would inhibit beta-endorphin release. Treatment with therapeutic agents that decrease oxidative stress, such as antibiotics and antioxidant phytotherapy, may function at least partially by increasing beta-endorphin and decreasing prostaglandin E2.

Delayed graft function. Influence on outcome and strategies for prevention.

Delayed graft function remains a prevalent problem in cadaveric renal transplantation that increases rejection, decreases graft and patient survival, increases the cost of transplantation, and complicates patient management. Although current medical and surgical strategies can reduce the incidence of DGF to 20% or less, newer therapies that focus on nonimmune and immune forms of renal injury are needed to improve outcomes further.

Oxidative stress in prostatic fluid of patients with chronic pelvic pain syndrome: correlation with gram positive bacterial growth and treatment response.

The etiology of chronic pelvic pain syndrome (CPPS)/chronic prostatitis category III remains unknown. Whereas a subset of men respond to antimicrobial therapy, gram positive bacteria isolated from expressed prostatic secretions (EPS) are often considered to be commensal rather than pathogenic. We wished to study oxidative stress as a marker of tissue injury and response in EPS of men with CPPS to determine whether infection with gram positive bacteria is associated with increased oxidative stress. A total of 300 EPS specimens from 100 men with CPPS were collected for microscopy, culture, and biochemical and molecular assays. Oxidant injury was measured by 8-isoprostane F2alpha (IsoP) levels and total antioxidant capacity as Trolox equivalents. Total RNA from EPS was used for gene expression of heme oxygenase-1 (HO-1) and granzyme B. The only bacteria found in EPS were gram positive. For our analysis, these men were classified as having chronic bacterial prostatitis (category II). IsoP levels (pg/mL) were highest in men with category II prostatitis (7315 +/- 1428) followed by nonbacterial prostatitis (category IIIa, 2043 +/- 561), prostatodynia (category IIIb, 319 +/- 81), and asymptomatic controls (298 +/- 99). IsoP levels decreased significantly after successful treatment with antibiotics or an antioxidant supplement (Prosta-Q). Antioxidant capacity was detected in 11 out of 18, 4 out of 16, and 1 out of 16 men tested with category II, IIIa, and IIIb prostatitis, respectively. No correlation was observed between IsoP levels and the number of white blood cells in EPS. HO-1 and granzyme B expression was highest in men with category II prostatitis than in men with either category III prostatitis or asymptomatic controls. On the basis of elevated oxidative stress, clinical response to antibiotics, and post-treatment reduction in oxidative stress, we conclude that gram positive bacteria in some men with CPPS may be pathogens. It is speculated that oxidative stress may be a key pathway in some men with CPPS that can be targeted with antioxidant therapy.

Detection of bacterial signal by 16S rRNA polymerase chain reaction in expressed prostatic secretions predicts response to antibiotic therapy in men with chronic pelvic pain syndrome.

PURPOSE: Some men with chronic pelvic pain syndrome (CPPS) have evidence of bacteria in their prostatic fluid (expressed prostatic secretions [EPS]) detected by 16S rRNA techniques. In this study, we correlate presence of bacterial signal with response to therapy. MATERIALS AND METHODS: EPS and first voided urine (VB1) from 47 men with CPPS were analyzed by polymerase chain reaction (PCR) for bacterial signal using universal primers specific for bacterial 16S rRNA. Signal was considered positive if found only in the EPS sample, or if at least 10x stronger in the EPS than in VB1. All patients were treated with antibiotic therapy. RESULTS: Thirty-three patients were category IIIa (nonbacterial prostatitis) and 14 were category IIIb (prostatodynia). Seventeen of the 33 category IIIa patients had positive localizing cultures for gram-positive bacteria. However, a positive bacterial signal was detected in 23 EPS samples by 16S rRNA PCR. This signal was found in 14 of 17 culture-positive patients, 7 of 16 of the remaining category IIIa patients, and 2 of 14 of category IIIb patients. No patient with negative bacterial signal improved with antibiotic therapy (negative predictive value 100%). Thirteen patients with positive bacterial signal improved with antibiotic therapy. CONCLUSIONS: In men with category III chronic prostatitis/CPPS, bacterial signal detected by PCR can help predict response to antimicrobial therapy.

Differences in acetylcholine but not choline in brain tissue fixed by freeze fixation or microwave heating.

Among the many rapidly metabolized compounds in the brain, acetylcholine is one of the most challenging to sample effectively due to its rapid synthesis, degradation and sequestration. To ascertain problems that invalidate sampling procedures two methods of tissue fixation, microwave heat inactivation and freeze fixation, were used for obtaining mice and rat brain samples, respectively. The data show that acetylcholine levels obtained by microwave fixation were much higher than those obtained by freeze fixation. Choline levels were not affected by the fixation method used. Microwave fixation results in more accurate assessment of acetylcholine levels than the freeze fixation method, even though the tissue fixation time was less than 1 s in both methods, because tissue integrity is maintained in the microwave fixation, but not during freeze fixation.

Expression of c-fos, c-jun and HSP70 mRNA in rat brain following high acceleration stress.

Rats exposed to high +Gz forces in a small animal centrifuge (SAC) exhibit loss of neuronal function (isoelectric EEG), termed G-induced loss of consciousness (G-LOC). This phenomenon is presumably due to a reduction in cerebral blood flow (CBF) or ischemia. Ischemia induces various metabolic and physiologic changes including expression of immediate early genes (IEGs) in the brain. Expression of IEGs have been suggested to be reliable markers for neuronal response to external stimuli or stress. In the present study expression of IEGs c-fos, c-jun and stress response gene HSP70 were measured in the brains of rats subjected to six 30 s exposures of +22.5Gz in a small animal centrifuge. The level of c-fos, HSP70 and beta-actin mRNA were measured by both Northern blot and RT-PCR. Expression of c-jun was measured only by RT-PCR. Expression of c-fos and c-jun was significantly stimulated at 0.5, 15, 30 and 60 min post-centrifugation. The level of HSP70 mRNA was significantly higher only at 60 and 180 min post-centrifugation. Measurement of metabolities showed a significant increase in lactate and a decrease in Cr-P level at 30 s and 15 min post-centrifugation, respectively. Lactate, but not Cr-P and ATP levels were restored to control levels by 60 min post-centrifugation. It is concluded that the transient expression of c-fos, c-jun and HSP70 mRNA is stimulated by repeated ischemic/reperfusion episodes induced by high acceleration stress.

Rat brain glucose and energy metabolites: effect of +Gz (head-to-foot inertial load) exposure in a small animal centrifuge.

A unique small animal centrifuge with on-line physiological monitoring and brain tissue collection (in < 1 s) capability was used to investigate the effect of increasing +Gz levels, exposure duration, number of exposures, and time course of metabolic changes in the rat brain. To determine the +Gz tolerance, rats were exposed to +7.5 to 25 Gz (30 s each) and EEG was monitored. G-induced loss of consciousness (G-LOC) defined as isoelectric EEG (I-EEG) occurred only at +22.5 and 25 Gz within 14.5 +/- 3 s. To study the effect of increasing +Gz levels on metabolism, rats were exposed to either 0.5 (control) or +7.5 to 25 Gz (30 s each), and brains were collected 1 min postcentrifugation by freeze fixation. A significant increase in lactate (> or = +7.5 Gz) and a decrease in glucose, creatine phosphate (Cr-P), and ATP levels were observed at +15 Gz and higher. The effect of exposure duration was investigated by exposing the rats to +22.5 Gz for 15-60 s. Brain lactate levels increased six-fold while glucose decreased (75%) following the 60-s exposure. The level of Cr-P and ATP decreased significantly after the 15- and 30-s exposures with no further changes at longer +Gz exposures. For time course studies, brains were collected both during (5-35 s) and after (1-15 min) a +25 Gz exposure. A significant decrease in Cr-P occurred within 5 s, but changes in glucose, ATP, and lactate required 15 s. All metabolites returned to control levels within 3 min, except lactate and adenosine, which required 15 min.(ABSTRACT TRUNCATED AT 250 WORDS)

c-fos and HSP70 gene expression in rat brains in high gravitation-induced cerebral ischemia.

Previous studies have shown that brief exposures of rodents to high gravitational forces (+Gz) in a specifically designed centrifuge cause global cerebral ischemia. In the present study, the effect of +Gz exposure to +22.5Gz for 15 to 60 s on c-fos and HSP70 gene expression was examined. Northern and RT-PCR analyses to total RNA isolated from brains of rats in different post-exposure times revealed a significant, time-dependent increase in the c-fos mRNA level which returned to near normal by 180 min. The HSP70 mRNA level was increased two-fold at 30 min post exposure, and remained elevated until 180 min. The transient stimulation of c-fos and HSP70 gene expression should serve as useful biomarkers for hypergravic stress on the brain. The present results should aid in design of future experiments in our understanding of the pathophysiology of the high +Gz challenges.

Multiple +Gz exposures cause brain edema in rats.

The most serious effect of high sustained +Gz (head-to-foot inertial load) known to occur in pilots of high performance aircraft is +Gz-induced loss of consciousness (G-LOC), which may result in pilot incapacitation and subsequent loss of life. G-LOC is believed to occur due to a critical reduction in cerebral blood flow (CBF). Recently, using a small animal centrifuge (SAC), we showed that +Gz exposure causes global cerebral ischemia in a rodent animal model. Since ischemia, depending upon the severity and duration, has been associated with increased brain water content or edema, the present study was undertaken. Rats were exposed to six exposures of either +25 Gz (30 s each) or +10 Gz (2 min each) in the SAC at +20 Gz.s-1 G onset rate. The appearance of G-LOC was monitored by the flattening of the electroencephalography (EEG) brain wave recording. G-LOC was observed at 101 +/- 46 and 19.2 +/- 5 s during +10 and +25 Gz exposures, respectively. The brains from these animals were removed 15 min to 24 h after the +Gz exposure and analyzed for edema formation (increase in the percentage of tissue water), metabolites, and cerebral blood volume (CBV). A significant decrease in glucose and an increase in lactate concentration were observed during +Gz exposure. Edema formation was observed 15 min after six exposures of either +10 or +25 Gz. A slight but significant decrease in CBV was also observed in rats exposed to six +10 Gz exposures. Edema formation was transient and resolved within 24 h. We concluded that multiple exposures of either +25 Gz, short duration or +10 Gz, long duration, that resulted in G-LOC, can cause cytotoxic brain edema which probably results from tissue hyperosmolality due to metabolic changes and accumulation of lactate during ischemia.

Effect of hypoxia and hyperoxia on human +Gz duration tolerance.

To determine the effects of varying inspired O2 on positive radial acceleration (+Gz; i.e., head-to-foot inertial load) duration tolerance, seven men were exposed to the +4.5- to +7.0-Gz simulated aerial combat maneuver (SACM) by use of the Armstrong Laboratory (Brooks Air Force Base) centrifuge. Exposures were repeated on different days while subjects breathed gas mixtures of fractional concentration of O2 in inspired air (FIO2) between 0.12 and 0.6. SACM duration tolerance was positively related to inspired O2 of FIO2 between 0.12 and 0.2 but was unchanged at FIO2 between 0.2 and 0.6. SACM exposure decreased arterial O2 saturation and increased heart rates; SACM-induced changes were additive to FIO2 effects. The positive relationship between blood lactate and SACM duration tolerance at all FIO2 indicated an anaerobic component. It is concluded that SACM duration tolerance is limited by reduced FIO2 but not enhanced by hyperoxia. Thus the aerobic component of +4.5- to +7.0-Gz SACM duration tolerance is much greater than previously believed.

Acceleration-induced effects on baboon blood chemistry.

Gravity-induced loss of consciousness (G-LOC) is known to have occurred in pilots since the early 1920's. Most of the research in this area has shown that G-LOC occurs due to a decrease in cerebral blood pressure and a concomitant reduction in brain perfusion. Since a reduction in cerebral blood flow can cause transient hypoxia, it is important to study the cerebral metabolism during high +Gz exposure. One component of these studies should include measurements of substrate availability and degradative products. In the present study, adult baboons were given multiple high +Gz exposures (2 to 6) using the Armstrong Laboratory human centrifuge. Venous blood was collected by an automatic syringe withdrawal pump before, during and after centrifuge exposures. The concentration of blood gases, glucose and lactate tended to decrease during the centrifuge exposure followed by an increase after the run. Total creatine kinase activity in serum was not significantly altered. These results suggest that during +Gz exposure, anaerobic glycolysis is stimulated resulting in elevated lactate production due to a reduction in cerebral blood flow (CBF). The elevated tissue lactate is released into the central circulation upon resumption of normal CBF (after the termination of centrifuge run). Therefore, the observed decrease in lactate concentration during the run may result from a lag in the release of tissue lactate into the blood due to a reduction in CBF. It is speculated that at high +6 Gz, G-LOC may occur as a protective response to reduce the brain metabolic rate, to maintain energy levels and to prevent severe cellular acidosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Stimulation of cAMP accumulation and superoxide production in human neutrophils and monocytes.

The effect of sodium fluoride (NaF) on superoxide generation and cyclic adenosine monophosphate (cAMP) levels in human neutrophils and monocytes was investigated. NaF (greater than 10 mM) stimulated superoxide (O2-) production in both cell types in a time dependent manner. NaF (0.5 to 20 mM) increased cAMP levels by 1.5- to 3.-fold in both neutrophils and monocytes. Increases in cAMP levels were time-dependent; the maximal level was attained within 5 minutes after the addition of NaF, and cAMP levels remained elevated for up to 10 minutes. Only high concentrations of NaF (10 and 20 mM) increased both cAMP levels and O2- production. Therefore, a direct role of cAMP in O2- generation is not likely. It is speculated that since NaF (greater than 10 mM) can complex with extracellular Ca++, and thus reduce free Ca++ concentration required for O2- generation, a NaF-dependent increase in cAMP may restore cytosolic free Ca++ by mobilizing intracellular stores of Ca++. Further, in view of the proposed involvement of a phosphorylation-dephosphorylation mechanism in the regulation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, we speculate that NaF, by inhibiting phosphoprotein phosphatase activity, may indirectly activate the NADPH oxidase system and thus superoxide generation.

Effect of NaF on cAMP accumulation, cAMP-dependent protein kinase activity in, and amylase secretion from, rat parotid gland cells.

Stimulation of amylase secretion from parotid glands by beta-adrenergic agonists is mediated by the activation of adenylate cyclase and the resultant increase in cellular cAMP. Since NaF is known to increase adenylate cyclase activity and cAMP accumulation in intact cells, we investigated whether it would stimulate amylase secretion from isolated rat parotid gland cells. The results provide evidence that the addition of NaF (0.01-10 mmol/L) increased cAMP concentration (1.5-2.8-fold) in, and amylase secretion (16-93%) from, isolated parotid gland acinar cells. NaF was found to increase cAMP-dependent protein kinase activity ratios (51-84%) in a concentration- and time-dependent manner. The data suggest that the stimulation of amylase secretion from parotid gland cells by NaF may be mediated by an increase in the cellular cAMP concentration, which exerts its effect, at least in part, by increasing the activity of cAMP-dependent protein kinase.

Stimulation of mucin release from rat submandibular salivary-gland cells by NaF.

The effect of NaF on cAMP accumulation, cAMP-dependent protein kinase activity (cAMP-dPK) ratios and [14C]-glucosamine-labelled mucin release from these isolated cells was investigated. NaF (0.01-5 mM) increased significantly the cellular cAMP concentration and cAMP-dPK activity ratios in a dose- and time-dependent manner. NaF (5.0 mM) increased [14C]-glucosamine-labelled mucin release in a time-dependent manner. Thus the stimulation of prelabelled mucin secretion by NaF is mediated by an increase in the cAMP concentration, which exerts its effect, at least partly, via the activation of cAMP-dPK activity.

Topical fluorides: effects on physiologic and biochemical processes.

The ingestion of fluoride from dentifrices or mouthrinses can contribute substantially to the total daily intake of the ion, even in communities that provide optimally fluoridated drinking water. It is concluded that the frequent and unsupervised use of these products by children six years of age or younger, especially those living in areas with water fluoridation, places them at risk of dental fluorosis. Recommendations to reduce the risk are presented. The use of 1.23% (12,300 ppm) APF gels, particularly in the absence of suctioning during the application and expectoration after the application, is associated with the swallowing of relatively large quantities of fluoride. The resulting increases in plasma fluoride levels may be sufficient to cause dental fluorosis, as judged by studies with laboratory animals, and a reduction in the kidney's ability to concentrate the urine, as judged by studies with both laboratory animals and humans. The epigastric distress experienced by some patients during or after APF gel applications appears to be due, at least in part, to a direct toxic effect of fluoride on the gastric mucosa. Data from studies with humans and laboratory animals indicate that there may also be associated changes in plasma and tissue cAMP levels, glucose metabolism, and salivary amylase secretion. There is an immediate need for the dissemination to the dental profession of standardized methods for gel application that will minimize the quantities of fluoride available for ingestion and systemic absorption.