The role of urine prostate cancer antigen 3 mRNA levels in the diagnosis of prostate cancer among Hong Kong Chinese patients.

OBJECTIVE: To establish and verify the utility of measuring urine prostate cancer antigen 3 (PCA3) mRNA levels in the diagnosis of prostate cancer among Hong Kong Chinese patients. DESIGN: Cross-sectional study. SETTING: Urology Unit of a regional hospital in Hong Kong. PATIENTS: This study was carried out in two parts. In the first part, 102 post-prostatic massage urine samples were collected from patients with known prostate cancer (38 patients) and controls (64 patients, with normal digital rectal examination and serum prostate-specific antigen <4 ng/mL). The urine levels of PCA3 and prostate-specific antigen mRNA were measured and the best cut-off point for differentiating cancer was determined. In the second part of the study, post-prostatic massage urine samples from 47 patients with clinically suspected prostate cancer were collected prior to prostate biopsy. The performance of PCA3 as a diagnostic aid for cancer was then assessed using the aforementioned cut-off value. RESULTS: In the first part of the study, the best cut-off for the PCA3 ratio (defined as the ratio of the Ct value of PCA3/PSA mRNA) was 1.127. Applying this cut-off to the 47 patients with clinically suspected prostate cancer and no history of previous prostate biopsy, the sensitivity and specificity of PCA3 for diagnosing prostate cancer were 71% and 92%, respectively. CONCLUSION: The post-prostatic massage urine PCA3 level shows utility for diagnosing prostate cancer in patients with elevated prostate-specific antigen levels that could facilitate decisions to undertake prostate biopsy and avoid unnecessary biopsies.

Is the decrease in the hypophysiotropic signal frequency normally observed during the luteal phase important for menstrual cyclicity in the primate?

The two phases of the ovulatory menstrual cycle of the primate are characterized by divergent activities of the GnRH pulse generator. During the luteal phase, LH pulse frequency is significantly reduced below that observed during the follicular phase. In this report we investigate whether the decrease in pulse frequency during the luteal phase is of physiological relevance to normal menstrual cyclicity. We have tested the effect of a pulsatile GnRH infusion given iv at hourly intervals for a period of 8-10 days during the luteal phase on the subsequent three to five cycles in eight female rhesus monkeys. Three of the eight animals received two treatment courses. Amounts of GnRH infused were 1.5 micrograms/pulse (n = 2 trials); 3.0 micrograms/pulse (n = 7); and 4.0 micrograms/pulse (n = 2). LH response to GnRH pulses of 1.5 and 3.0 micrograms resembled spontaneous LH pulses observed during the luteal phase. During the GnRH infusion period, the monkeys were fitted with a primate vest and tethered. Eleven control experiments were performed in these monkeys under similar conditions. GnRH therapy during the luteal phase affected subsequent cycles significantly, while no differences were observed in the control experiments. Overall mean follicular phase length in the control cycle was 13.4 days; it was significantly increased (P less than 0.005) in all post-GnRH treatment cycles to reach 34.4 (+/- 10.9), 43.9 (+/- 12.7), 40.4 (+/- 13.0), and 23.1 (+/- 4.8) days (+/- SE) in the first to fourth post-GnRH cycles, respectively. Progesterone secretion was significantly lower (P less than 0.05) in the first two post-GnRH cycles than in the control cycles: progesterone, 46.4 (+/- 2.1) in all control cycles, decreased to 27.7 (+/- 3.7), 24.8 (+/- 4.3), 34.0 (+/- 5.4), and 32.0 (+/- 6.5) surface units (+/- SE) from the first to fourth post-GnRH cycles, respectively, while luteal phase length remained relatively unchanged. The data indicate that significant disturbances in the menstrual cycle of the rhesus monkey follow imposed changes in the normal frequency pattern of the GnRH hypophysiotropic signal during the luteal phase and suggest that the naturally occurring slowing of GnRH-LH pulse frequency during the luteal phase is a relevant phenomenon in the sequence of events which control menstrual cyclicity.

Reduced frequency of pulsatile luteinizing hormone secretion in the luteal phase of the rhesus monkey. Involvement of endogenous opiates.

Pulsatile secretion of LH in women has been shown to vary during the menstrual cycle. LH pulse frequency during the luteal phase is markedly reduced compared to that in the follicular phase. The objectives of the present study were to determine if similar changes in pulsatile LH secretion occur in the monkey, and whether endogenous opiates are involved in producing these changes. In order to document if LH pulse frequency is reduced in the nonhuman primate luteal phase, serial blood samples were collected from 10 rhesus monkeys at 15-min intervals for 6 h at 3 different times of the luteal phase (early, mid-, and late). This pattern of secretion was contrasted to that observed during the ensuing early follicular phase. LH pulse frequency during the luteal phase was significantly reduced compared to the early follicular phase. Mean pulse frequency (+/- SE) was 0.84 +/- 0.16 pulses/6 h in the luteal phase vs. 2.99 +/- 0.58 pulses/6 h in the early follicular phase. When endogenous opioid activity was blocked during the luteal phase by a 5-h continuous infusion of naloxone (2 mg/h), an opiate antagonist, LH pulse frequency was increased to 2.48 +/- 0.25 pulses/5 h. This frequency was markedly different from the frequency of 0.85 +/- 0.17 pulses/5 h observed in the control period which immediately preceeded the naloxone infusion. The mean amplitude of the LH pulses in the luteal phase, which was significantly greater than that observed in the early follicular phase (20.9 +/- 1.9 ng/ml and 11.7 +/- 0.3 ng/ml) was not affected by naloxone (23.5 +/- 2.4 ng/ml vs. 25.3 +/- 1.9 ng/ml). Infusion of naloxone for longer periods (9 h) in 3 additional monkeys caused an increase in LH pulse frequency which was maintained in 2 of the monkeys, whereas the third animal exhibited only an acute response (a single pulse). These results indicate that the reduction in LH pulse frequency that occurs in the luteal phase of the rhesus menstrual cycle is an event in which endogenous opiates participate. Our previous finding that beta-endorphin release from neurons in the median eminence is stimulated during the luteal phase of the monkey, together with the present results, suggest that beta-endorphin functions as a modulator of pulsatile LH secretion in the primate menstrual cycle.

Effects and site of action of morphine on gonadotropin secretion in the female rhesus monkey.

The effects of morphine on gonadotropin secretion, and the site of its action, were tested in female rhesus monkeys. In Exp 1, morphine sulfate (3, 6, or 9 mg iv) was injected into ovariectomized monkeys, and its effects on tonic (pulsatile) LH and FSH secretion were examined. Administration of morphine (9 mg) resulted in a significant decrease in circulating LH and FSH, which lasted for 4-5 h. Exp 2 was performed to evaluate the site of action of morphine, whether hypophyseal or suprahypophyseal. The effects of morphine (6, 9, or 12 mg) on the LH response to GnRH pulses were evaluated in stalk-sectioned monkeys, in which gonadotropin secretion had been restored by long term pulsatile infusion of GnRH. LH responses to GnRH were not significantly altered by morphine. Exp 3 was performed to determine the effects of morphine on the estrogen-induced LH surge. Estradiol benzoate (330 micrograms in oil) was administered on days 2-5 of the menstrual cycle to nine animals. Four of these also were injected with 9 mg morphine at 5-h intervals for 40 h. Four of the five control and three of the four morphine-treated monkeys showed similar LH surges. The results demonstrate that, in the monkey, opiates inhibit tonic (pulsatile) gonadotropin secretion, most probably by acting at a suprahypophyseal site. In contrast, morphine does not alter the estradiol-induced LH surge, a result that differs from that seen in lower species and that may be related to differences in estradiol positive feedback characteristics.

Derivation of a prediction rule for posttraumatic organ failure using plasma DNA and other variables.

The early identification of patients at high risk of developing posttraumatic organ failure would allow preventive therapies to be studied. In this study, highly sensitive and specific guidelines for the early prediction of posttraumatic organ failure (OF) and multiple organ dysfunction syndrome (MODS) using cell-free (plasma) DNA and other predictors of posttraumatic complications were derived. As plasma DNA increases after injury and may be used to predict acute lung injury (ALI), we hypothesized that in combination with other predictors it would predict the later development of OF and MODS. Eighty-three patients (69 males; median age, 36 years) were studied as a consequence of major trauma within 3.5 hours of injury (median time to sampling and assessment, 60 min). Plasma DNA was measured using a real-time, quantitative, polymerase chain reaction assay for the beta-globin gene. OF and MODS occurred in 20/83 (24%) and 9/79 (11%) cases, respectively. At selected cutoff points, the sensitivity of plasma DNA for predicting OF and MODS ranged from 50% to 100%, specificity ranged from 74% to 95%, and the likelihood ratio ranged from 3.89 to 10.50. Other variables studied included serum albumin, creatine kinase, aspartate transaminase, lactate dehydrogenase, leukocyte count, hematocrit, injury severity score, maximal abbreviated injury score, and shock index. Using a classification and regression tree, plasma DNA and aspartate transaminase at optimal cutoffs predicted OF and MODS with an overall correct classification of 93% and 87%, respectively.

Increased plasma free cyclic-AMP levels following major trauma and their relevance to the immune response.

BACKGROUND: Following injury, neutrophil mobilisation is an important element of the immune response. The ideal features of an agent responsible for this mobilisation would include the ability to mobilise neutrophils without activating them, and rapid reversibility. This study investigates the hypothesis that raised levels of plasma adrenaline following trauma act via cyclic adenosine monophosphate (cAMP) to mobilise neutrophils, and measures the amount of cAMP extruded from cells into the plasma following injury. METHODS: 20-ml samples of venous blood were drawn from 34 trauma patients within 3 h of injury and divided between three sample tubes: (1) ethylene diamine tetra-acetic acid anticoagulant (EDTA) for full blood count; (2) cooled EDTA for cAMP levels; and (3) cooled lithium heparin for catecholamines. The latter two tubes were immediately centrifuged at low temperature and the supernatant plasma deep frozen pending analysis. Adrenaline was measured using high pressure liquid chromatography (HPLC) and cAMP measured by an enzyme immunoassay technique. RESULTS: 34 patients, six of whom had sustained minor trauma (ISS 1-8), 12 moderate trauma (ISS 9-15) and 16, major trauma (ISS 16 and above) were studied. Median age was 39 years (range 16-77) and 30 patients were male. Plasma adrenaline levels were available for 28 of the patients. Plasma free cAMP levels were significantly raised in patients with major trauma (P < 0.006). There were positive correlations between the plasma levels of cAMP and adrenaline (rho 0.660, P = 0.011), adrenaline and neutrophil count (rho 0.654, P = 0.01) and cAMP and neutrophil count (rho 0.508, P = 0.013). CONCLUSIONS: Evidence is presented of the possible inter-relationships between neutrophil counts, adrenaline levels and cAMP levels following injury, supporting the proposition that neutrophil mobilisation pathways are activated early via beta-adrenergic stimulation.

Role of monocyte L-selectin in the development of post-traumatic organ failure.

The vascular leucocyte adhesion molecule, L-selectin, plays an important early role in monocyte trafficking at sites of inflammation, a process which leads to the development of inflammatory organ failure. In this prospective observational study, we investigate whether early numerical and functional changes in circulating monocytes, expression of monocyte L-selectin (CD62L) and monocyte:neutrophil L-selectin ratios are related to the subsequent development of post-traumatic organ failure (OF) and multiple organ dysfunction syndrome (MODS). Monocyte counts and cell surface L-selectin were measured by an automated cell counter and flow cytometry, respectively. Of 164 trauma patients admitted to a university emergency department resuscitation room, 64 had multiple injuries, 51 developed OF, 20 developed MODS and 21 died. Early monocyte counts in patients with multiple injuries were lower in those who developed MODS (0.44 x 10(9)/l) compared with those who did not (0.60 x 10(9)/l; P=0.024). Monocyte L-selectin mean channel fluorescence increased with injury severity and was highest in those who developed MODS (P=0.033). In the sub-group of patients with multiple injuries, L-selectin mean channel fluorescence was also greater in those patients who developed MODS compared with patients who did not develop MODS (P=0.042). The monocyte to neutrophil count ratio also decreased with injury severity (P=0.006). Using optimal cut off values for L-selectin mean channel, fluorescence, the positive and negative predictive values for OF was 43.5 and 91.4%, respectively and for MODS it was 25.4 and 92.9%, respectively. Alterations in early circulating monocyte counts and L-selectin expression after injury are related to the development of post-traumatic organ failure and suggest an area in the inflammatory pathway that may be influenced by L-selectin blockade.

The Prostate Health Index in predicting initial prostate biopsy outcomes in Asian men with prostate-specific antigen levels of 4-10 ng/mL.

PURPOSE: To investigate the role of the Prostate Health Index (phi) in prostate cancer (PCa) detection in patients with a prostate-specific antigen (PSA) level of 4-10 ng/mL receiving their first prostatic biopsy in an Asian population. METHODS: This was a retrospective study of archived serum samples from patients enlisted in our tissue bank. Patients over 50 years old, with PSA level of 4-10 ng/mL, a negative digital rectal examination, and received their first prostatic biopsy between April 2008 and April 2013, were recruited. The serum sample collected before biopsy was retrieved for the measurement of various PSA derivatives and the phi value was calculated for each patient. The performance of these parameters in predicting the prostatic biopsy results was assessed. RESULTS: Two hundred and thirty consecutive patients, with 21 (9.13 %) diagnosed with PCa, were recruited for this study. Statistically significant differences between PCa patients and non-PCa patients were found for total PSA, PSA density, [-2]proPSA (p2PSA), free-to-total PSA ratio (%fPSA), p2PSA-to-free PSA ratio (%p2PSA), and phi. The areas under the curve of the receiver operating characteristic curve for total PSA, PSA density, %fPSA, %p2PSA, and phi were 0.547, 0.634, 0.654, 0.768, and 0.781, respectively. The phi was the best predictor of the prostatic biopsies results. At a sensitivity of 90 %, the use of the phi could have avoided unnecessary biopsies in 104 (45.2 %) patients. CONCLUSIONS: Use of the phi could improve the accuracy of PCa detection in patients with an elevated PSA level and thus avoid unnecessary prostatic biopsies.

Early role of neutrophil L-selectin in posttraumatic acute lung injury.

OBJECTIVE: To investigate whether early numerical and functional changes in circulating neutrophils and expression of neutrophil L-selectin and soluble L-selectin are related to the subsequent development of posttraumatic acute lung injury (ALI), the systemic inflammatory response syndrome, sepsis, and organ failure. DESIGN: Prospective study of whole blood and plasma samples to assess numerical and functional changes in circulating neutrophils and in soluble L-selectin. SETTING: The emergency department of a university hospital. PATIENTS: A total of 147 patients admitted to the resuscitation room after trauma were compared with 69 control subjects. Ten patients developed ALI. LABORATORY ANALYSIS: Flow cytometry of whole blood and ELISA of plasma. RESULTS: Total leukocyte and neutrophil counts, expression of L-selectin, and the ratio of neutrophil to plasma L-selectin increased with injury and were highest in those who developed ALI. Soluble L-selectin decreased with injury severity and was lowest in those who developed ALI. CONCLUSIONS: Early changes in the average expression of L-selectin per cell do not correlate with the development of subsequent posttraumatic ALI. However, the development of ALI is related to the total expression of L-selectin in the neutrophil mass, and the most striking association is in those with lower concentrations of plasma L-selectin.

Transplanted gonadotropin-releasing hormone neurons promote pulsatile luteinizing hormone secretion in congenitally hypogonadal (hpg) male mice.

Congenitally hypogonadal (hpg) male mice are unable to synthesize biologically active gonadotropin-releasing hormone (GnRH). Implantation of normal fetal preoptic area tissue containing GnRH neurons into the third ventricle of adult hpg males significantly elevates pituitary levels of luteinizing hormone (LH) and corrects their hypogonadism. In all responding animals, immunoreactive GnRH neurons within the transplant innervate the median eminence of the host. To assess whether gonadal recovery in hpg hosts results from pulsatile secretion of GnRH from grafted neurons, we compared the pattern of variation in plasma LH levels in 19 hpg graft recipients with testicular growth to that of 10 normal adult mice. All animals were castrated prior to receiving an indwelling catheter in the jugular vein. Sequential blood samples were collected (t = 10 min) and assayed for LH. Pulsatile LH secretion was seen in 11 of 19 hpg hosts and in all control mice. While there was great variability between individual animals, measures of baseline LH, LH pulse amplitude and duration, interpulse interval, and LH pulse frequency revealed no difference between hpg graft recipients and normal castrates in their LH pulse pattern. Immunocytochemical analysis of the brain in hpg hosts suggested no correlation between any parameter of pulse activity and individual differences in GnRH cell number or GnRH fiber outgrowth into the median eminence. Sources of variation in LH secretion among graft recipients, and between hpg hosts and normal mice, are discussed. We suggest that transplanted GnRH neurons are capable of integration into a GnRH 'pulse generator' which can support a near-normal pattern of pulsatile LH secretion, leading to testicular growth and steroid production.