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We established an ultrasensitive method for identifying multiple enzymes in biological samples by using a multiplexed microdevice-based single-molecule enzymatic assay. We used a paradigm in which we "count" the number of enzyme molecules by profiling their single enzyme activity characteristics toward multiple substrates. In this proof-of-concept study of the single enzyme activity-based protein profiling (SEAP), we were able to detect the activities of various phosphoric ester-hydrolyzing enzymes such as alkaline phosphatases, tyrosine phosphatases, and ectonucleotide pyrophosphatases in blood samples at the single-molecule level and in a subtype-discriminating manner, demonstrating its potential usefulness for the diagnosis of diseases based on ultrasensitive detection of enzymes., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)

Background/aim: For prostate cancer, positive surgical margins are considered an important predictor of biochemical recurrence. However, biochemical recurrence is observed in approximately 20% of cases, even with negative surgical margins, and some cases require salvage therapy. The elevated expression of low-molecular-weight protein tyrosine phosphatase (LMW-PTP, MW 18 kDa) is associated with a poor prognosis of certain cancers. In this study, we investigated whether the LMW-PTP expression levels could be used as a biomarker of recurrence in prostate cancer with negative surgical margins., Materials and Methods: The subjects of this retrospective study were 119 patients who underwent total prostatectomy with negative resection margins. LMW-PTP expression was categorized either as a high-expression group or as a low-expression group bye two pathologists. Subsequently, we examined the relationship between LMW-PTP expression levels and clinicopathological factors including biochemical recurrence., Results: Evaluation of the immunostained samples by two pathologists was highly reliable, with an Intraclass correlation (ICC) score for two distinct measurements of 0.77 and 0.98, respectively. Seventy-three patients (61.3%) were placed in the LMW-PTP high expression group; and 46 patients (38.7%) were placed in the low expression group. The log-rank test revealed early biochemical recurrence in the high LMW-PTP expression group (p=0.0001). In addition, pathological T stage (p=0.004), lymphatic invasion (p=0.0456), Ki-67 labeling index (p=0.0002), and biochemical recurrence (p<0.0001) were more frequently identified in the LMW-PTP high expression group. Furthermore, multivariate analyses revealed that a high LMW-PTP expression level was an independent prognostic factor for biochemical recurrence (HR=3.14, 95% CI=1.37-8.07, p=0.0057). In addition, Ki-67 labeling indices were significantly higher in the high-expression group compared to the low-expression group (p<0.0001)., Conclusion: LMW-PTP can be assessed using a single immunostaining protocol in a highly reproducible fashion. Tt may, thus, be applied clinically to establish the required postoperative follow-up period and determine the necessity for salvage therapy in cases of prostate cancer with negative surgical margins. LMW-PTP has the potential to be a highly useful prognostic biomarker and a therapeutic target in conjunction with other factors, such as the Gleason Score, the pathological T stage and the PSA level., (Copyright© 2019, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Mycobacterium avium ssp. paratuberculosis (Map) is the etiological agent of Paratuberculosis in ruminants. Protein tyrosine phosphatase A (PtpA) and protein kinase G (PknG) are secreted proteins necessary for the survival of the pathogen within macrophages. In this study we analyzed if Map was able to grow within astrocytes and investigated on the presence of antibodies against PtpA and PknG proteins in MS and NMOSD patients by ELISA. Map was unable to proliferate in astrocytes after of 72 h post-infection, but we observed a high level of antibodies against both virulence factors, suggesting that these patients have been exposed/infected with Map., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objective: We evaluated the prevalence of glutamic acid decarboxylase (GADA) and tyrosine phosphatase-protein antibodies (IA2A), their titers and their relation to first phase insulin response (FPIR) and glucose tolerance in autoimmune thyroid diseases (ATDs) patients., Subjects and Methods: Graves' disease (GD; n = 181) and Hashimoto's thyroiditis (HT; n = 143) patients in addition to healthy controls (n = 93) were studied. Secondly, FPIR and oral glucose tolerance tests (OGTT) were performed in 11 anti-pancreatic islet-cell (+) and in 20 anti-pancreatic-cell (-) patients., Results: There was a non significant trend for higher prevalence of GADA positivity in GD vs HT (7.2% vs 2% p = 0.06), but the GADA titers were higher in HT. We also did not find a significant difference in IA2 prevalence (0.7% vs 0.0%) between these two groups or compared to the control group. In the subsequent analysis, low FPIR was found in 10% of these patients but without statistical difference for OGTT between pancreatic antibody-positive and -negative patients., Conclusion: A trend for greater prevalence of GADA was observed for GD patients than for HT or control. However, the titers of these autoantibodies were higher in HT patients, but there was no significant relation to insulin secretion and glucose tolerance at that moment and stage of autoimmune diseases.

Prostate cancer remains a leading cause of cancer death in Europe and the United States and is an emerging problem in Asia despite significant improvements in available treatments over the last few decades. Androgen deprivation therapy (ADT) has been the core treatment of advance-staged disease since the discovery of prostate cancer's androgen dependence in 1941 by Huggins et al. [1] Options for initial medical treatment include gonadotropin-releasing hormone analogues such as leuprolide (LHRH agonist) and degarelix (LHRH antagonist) and androgen receptor (AR) binding agents such as bicalutamide. Although most patients will initially respond to either surgical or medical castration, there is almost always progression to castration-resistant prostate cancer (CRPC) necessitating treatment with more novel agents. [2] However, even drugs such as abiraterone and enzalutamide, two next-generation agents used commonly in metastatic CRPC, have failed to demonstrate persistent efficacy in most patients. [3] ,[4].

Aim: Lycopene is a member of the carotenoid family and has strong anti-oxidant properties. Lycopene occurs in tomato-based food products primarily as an all-E isomer (80-97%),but its Z-isomers accounts for 79 to 88% of total lycopene in benign or malignant prostate tissues, while the specific biological functions of Z-isomers are still not clarified at present. This study was to examine the bioactive potency of Z-isomers on benign prostatic hyperplasia (BPH) in mice and to make a comparison of effective inhibition between Z-isomers and all-E isomer., Method: Mice were divided into the Saline group, Vehicle control group and testosterone propionate induced BPH mice group (BPH model group, vehicle BPH model group, lycopene treated (5 mg/kg and 2.5 mg/kg), Z-isomers (57%) treated, Z-isomers (86%) treated, finasteride treated). The drugs were orally administered once a day consecutively for 30 days. The inhibitory effects on BPH of all-E lycopene and Z-isomers were evaluated by prostatic index, prostatic acid phosphatase (PAP), estradiol, testosterone and dihydrotestosterone (DHT) levels in serum and histopathology examination., Results: Compared with the BPH model group, E/Z isomers exhibited significant differences in prostatic index, PAP, estradiol, testosterone and DHT levels in serum and similar histological aspects observed in the mice of the control group. The present research also shows that Z-isomers may be more potent inhibitors than all-E isomers in BPH treatment., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Ductal adenocarcinoma of the prostate is considered to be a rare variant of prostatic adenocarcinoma when compared to the more common acinar adenocarcinoma. We report here a case of ductal adenocarcinoma of the prostate in a 68-year old man who presented with complaints of abdominal pain, retention of urine and hematuria of one month duration. Clinical examination showed prostatomegaly. The serum Prostate Specific Antigen (PSA) value was raised to 79ng/mL. Histopathological and immunohistochemical evaluation of resected specimen of prostate revealed ductal adenocarcinoma of the prostate. The patient was lost to follow up and presented four years after the initial diagnosis with metastasis to the bone and testis. Though prostatic cancers have the ability for wide spread dissemination, metastasis to testis is rare. Immunohistochemical staining with PSA and Prostatic Acid Phosphatase (PAP) can help in establishing prostatic nature of the neoplasm. We are reporting this case because of the rarity of metastasis of prostatic carcinoma to testis and for stressing the need for keeping in mind the possibility of metastatic carcinoma also while dealing with testicular tumors.

Recent evidence has shown that plasma fibrinogen, a major cardiovascular risk factor, interacts with the erythrocyte membrane and acts to influence blood flow via erythrocyte nitric oxide (NO) modulation. In the present in-vitro study, whole blood samples were harvested from healthy subjects and aliquots were incubated in the absence (control aliquots) and presence of fibrinogen at different degrees of band 3 phosphorylation, and the erythrocyte deformability was determined. The present study shows that in the presence of higher fibrinogen concentrations, similar to those found in inflammatory conditions, erythrocyte deformability is increased only when band 3 is dephosphorylated by the presence of syk inhibitor and at low shear stress. On the contrary, no changes were verified in the presence of fibrinogen when band 3 is allowed to be phosphorylated by inhibiting the phosphotyrosine phosphatase enzyme activity with calpeptin. We also observed that the presence of fibrinogen at higher concentration does not induce changes in erythrocyte deformability in the absence of modulators of the band 3 phosphorylation degree. However, the mechanisms by which fibrinogen signalling modulates erythrocyte function remain to be clarified and are currently under study.

Adenoid cystic carcinoma is an unusual histological variant of prostatic carcinoma. Because of its rarity, the natural history of this tumor is not known. Here we report this rare entity in a 62-year-old man who presented with symptoms of urinary tract obstruction. Digital rectal examination and ultrasonography (USG) showed an enlarged hard nodular prostate. Serum prostate-specific antigen (PSA) and prostatic acid phosphate levels were found to be within the normal range. Transrectal ultrasound-guided 12 core biopsies of prostate showed morphological features of an adenoid cystic carcinoma in 8 cores (bilateral, mid and base) on histopathological examination. Immunohistochemistry performed for PSA on paraffin section was negative. After diagnosis, bilateral orchidectomy was performed, and hormonal therapy was started in the form of androgen receptor blocker. The patient was clinically stable during a limited follow up of six months., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

Prostate specific antigen (PSA) is still the most useful tool to select the population requiring prostatebiopsy. The main downsides of PSA are an inadequate sensitivity to be used in screening and a low specificity for cancer detection. So far, a limited value for PSA derivates (velocity, density, free, proisoforms and doubling time) has been recognised. We present a short review of the literature describing a selection of the most promising alternatives to PSA being studied currently: PCA3, serum kallikreins, serum detectable prostate specific membrane antigen, the nuclear matrix protein EPCA, EPCA-2, prostatic acid phosphatase, urine detectable GSTP1, anti-AMACR antibodies, sarcosine, plasminogen activating urokinase, IGFBP, TGF beta 1,PSP94, IL6, plasmatic DNA, serum autoantibodies, neuroendocrine markers, proteomic analysis.

Introduction: Pumpkins are thought to be useful in the management of benign prostatic hyperplasia (BPH). The ability of a 15% Telfairia occidentalis seeds incorporated diet to inhibit hormonal induction of BPH in rats was studied., Materials and Methods: Twenty male Wistar rats were divided into 4 equal groups - one test group and three control groups. The test group was placed on the test diet and was given subcutaneous injections of dihydrotestosterone (DHT) and estradiol valerate (ratio 10:1) every other day for 28 days. One control group, 'no test diet' (ND) group, received the hormones, but was placed on a normal diet. The other two control groups, 'no hormone' (NH) and 'no hormone/test diet' (NHD), received subcutaneous olive oil (vehicle) for the same duration and were placed on the test and normal diets, respectively. Markers of BPH and hormone profile were determined using standard methods., Results: The mean relative prostate weight (×10(3)) was reduced in the test group (3.6 ± 0.2) relative to the ND group (4.0 ± 0.4). The protein content (mg/tissue) of the rats' prostates decreased significantly (p < 0.05) from 68.3 ± 2.7 in the ND group to 43.4 ± 3.9 in the test group. Serum prostatic acid phosphatase levels (U/l) decreased significantly (p < 0.05) from 4.8 ± 0.4 in the ND group to 4.0 ± 0.9 in the test group. Histological findings corroborate these data. The testosterone:estradiol ratio (×10(3)) was significantly (p < 0.05) increased from 7.1 ± 0.1 in the ND group to 8.4 ± 0.4 in the test group., Conclusion: The test diet inhibited the induction of BPH in rats and may act by increasing the testosterone:estradiol ratio., (Copyright © 2011 S. Karger AG, Basel.)

Background: fulminant type 1 diabetes mellitus (T1DM) is characterized by abrupt onset of hyperglycemia and rapid progression to ketoacidosis. This study aimed at examining the clinical and autoimmunity features of fulminant T1DM in Chinese., Methods: a total of 24 patients with fulminant T1DM and 48 classical autoimmune T1DM patients with acute onset of ketoacidosis were recruited. Anthropometric and biochemical parameters were tested. Serum antibodies against glutamic acid decarboxylase 65, tyrosine phosphatase-2 and zinc transporter 8 were measured, and human leukocyte antigen-DQ haplotypes were determined. Peripheral glutamic acid decarboxylase 65-specific T-cell responses in some subjects were determined., Results: compared with autoimmune T1DM patients, patients with fulminant T1DM displayed more flu-like and gastrointestinal symptoms, and had significantly higher concentrations of plasma glucose, more severe ketoacidosis, very low levels of pancreatic β-cell reserve, but only slightly increased haemoglobin A(1c) levels. In some patients, the disease onset was associated with drug-related hypersensitivity, pregnancy, or positive serum IgM against viruses. Forty percent (8/20) had low titres of autoantibodies against at least one of the islet autoantigens tested. Three out of six patients had positive glutamic acid decarboxylase-reactive Th1 responses. The frequency of human leukocyte antigen -DQA1*0102-DQB1*0601 haplotype was significantly higher in patients with fulminant T1DM., Conclusions: fulminant T1DM is a distinct entity with unique clinical characteristics and may be mediated by multiple factors, including viral infection, pregnancy, and drug sensitivity syndrome, among others, in the presence of humoral and/or cellular autoimmunity and susceptible genetic background., (2010 John Wiley & Sons, Ltd.)

Protein tyrosine phosphatase interacting protein 51 (PTPIP51) was identified as an in vitro interacting partner of protein tyrosine phosphatase 1B (PTP1B) and T-cell protein tyrosine phosphatase (TCPTP). The full-length form of PTPIP51 encompasses 470aas and has a molecular weight of 52kDa. The physiological function is poorly understood but an involvement in differentiation processes and apoptosis has been suggested. Preliminary observations suggested differences in PTPIP51 expression in blood cells. To analyze a possible involvement of PTPIP51 in hematopoietic processes, we studied its expression in samples of peripheral venous blood (PVB), umbilical cord blood (UCB) and human bone marrow (HBM). In both, PVB and UCB PTPIP51 expression was restricted to neutrophil granulocytes. In HBM samples, besides in mature neutrophil ganulocytes PTPIP51 protein and mRNA was present in myeloid precursor cells of neutrophils. The expression of PTPIP51 in neutrophil granulocytes was corroborated by immunoblot analysis exhibiting different molecular weight forms of PTPIP51 protein. Anti-peptide antibodies, identifying specific regions of the PTPIP51 protein (C-terminus, N-terminus and aas114-129) revealed a distinct isoform expression pattern in neutrophil granulocytes of different sources. In PVB and UCB neutrophil granulocytes reacted positive for all three peptide antibodies. In contrast, neutrophils of HBM express solely an N-terminal variant of PTPIP51 protein, lacking the C-terminal and aas114-129 sequence. Immunocytochemical results displayed a strict co-localization of PTPIP51 and PTP1B in PVB and UCB. The interaction of both proteins was verified by a proximity ligation assay. Neither proliferating cells, as identified by PCNA immunostaining, nor apoptotic cells, labeled by TUNEL assay, displayed an immunoreactivity for PTPIP51 in HBM. In fact, PTPIP51 expression was restricted to myeloid precursor cells undergoing differentiation. In blood cells therefore, PTPIP51 expression is restricted to differentiating and mature neutrophil granulocytes., (2010 Elsevier Inc. All rights reserved.)

The effect of different doses of cadmium [CD] on some biochemical, hormonal and histopathological parameters of the liver, kidney and testes of the Wistar rate were investigated. Cadmium in the dose range 0-40 mg/kg while causing a time-and dose-dependent decrease of the basal serum levels of alkaline phosphatase [ALP] also caused a dose-dependent increase in the serum concentration of the acid and prostatic acid phosphatases. The value of the ALP changed from 148.7+/-1.0 IU/L in the control to 53.7+/-0.098 at 40 mg/kg of cadmium. While the ACP and ACPT changed from 32.6+/-0.72 and 7 Units in the control to 54 and 17 units respectively at 40 mg/kg of CD. Furthermore cadmium also caused positively correlated dose-and time-dependent destruction of the histology of the liver, kidney and testes. These were characterized by vascular congestion, vacuolation, destruction of the seminal epithelial layers, focal necrosis of nucleus, oedema of the seminal epithelia layers, focal necrosis of nucleus, oedema of the seminiferous tubules and reduction of spermatogenesis. CD also caused granular and eosinophilic cytoplasm, enlargement of sinusoids with kupffer cells, haemorrhage and apoptosis of cells. Finally pre-treatment with vitamin C [0.0015/kg], vitamin E [1.51/g] and selenium [0.25 mg] which on their own had little or no effects on the serum basal phosphatases, hormonal and histological stability caused a reversal of the cadmium-induced biochemical, hormonal and histological toxicities of the liver, kidney and testes. These results may be explained by the oxidational/antioxidational properties of these xenobiotics and their mechanisms of actions.

The effects of different doses of ammonium metavanadate on the biochemical, haematological, hormonal and histopathological parameters of stilbesterol treated female Wistar rats were investigated. Ammonium metavanadate in the dose range 0-6 mg/kg caused a bi-phasic and time-dependent response on the acid [total and prostate] phosphatase. Furthermore ammonium metavanadate caused a dose-dependent inhibition of the serum alkaline phosphatases. The maximal inhibitory response at 5mg/kg of ammonium metavanadate was 40.0+/-1.69 compared to 65.0+/-0.94 control values. Ammonium metavanadate also caused a positively correlated biphasic response in the serum female hormonal concentrations with an initial increase, followed by a time-dependent decrease in the serum values of luteinizing (LH), follicle stimulating hormone (FSH) and prolactin. Furthermore ammonium metavanadate also caused time-and dose-dependent effects on the haematological parameters. The effects were biphasic-increase within 72 hours and a reduction in the values of haemoglobin and packed cell volume within 7-28 days. The white blood count and lymphocyte counts were also reduced [P<0.05] significantly. However the neutrophil counts were increased dose-and time-dependently. Finally, ammonium metavanadate caused a dose-dependent destruction of the liver and female reproductive organs namely the uterus, ovary and fallopian tubes. These were characterized by necrosis, oedema, eosinophilic deposits and vacuolation. These results may be explained by the oxidative effects caused by the free oxygen [O2] radical generated by the metavanadate ions.

Prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) are glycoproteins secreted by prostate epithelial cells, and have a long clinical history of use as serum biomarkers of prostate cancers. These two proteins are present at significantly higher concentrations in seminal plasma, making this proximal fluid of the prostate a good source for purifying enough protein for characterization of prostate disease associated changes in glycan structures. With the use of seminal fluid samples representative of normal control, benign prostatic disease and prostate cancers, PAP and PSA were enriched by thiophilic absorption chromatography. Released N-linked glycan constituents from both proteins were analyzed by a combination of normal phase HPLC and MALDI-TOF spectrometry. For PSA, 40 putative glycoforms were determined, and 21 glycoforms were determined for PAP. PAP glycans were further analyzed with a hybrid triple quadrupole/linear ion trap mass spectrometer to assign specific glycoform classes to each of the three N-linked sites. The glycans identified in these studies will allow for more defined targeting of prostate disease-specific changes for PAP, PSA and other secreted prostatic glycoproteins.

Objective: Genetic differences in the activity of phosphotyrosine phosphatases between mother and embryo could result in a differential activation of signals induced by growth factors in the two sides of placenta. Previous observations suggest that this may have important effects on intrauterine development and survival. The aim of the present study is to confirm previous observations and show new data., Study Design: We have studied 573 mother/newborn pairs, 169 wife/husband couples with repeated spontaneous abortion and 34 fertile wife/husband couples, Results: In mother/newborn pairs, the analysis of joint mother/infant ACP1 distribution has shown a deficit of pairs with the mother having low ACP1 S isoform concentration and the infant having high S isoform concentration, and an excess of pairs with the mother having high S isoform concentration and the infant having low S isoform concentration. In RSA couples there is an excess of couples in which the wife has low S isoform concentration and the husband has high S isoform concentration and a deficit of couples in which the wife has high S isoform concentration and the husband has low S isoform concentration. In fertile couples the pattern is reversed., Conclusion: The data suggest that when the mother to fetus S isoform concentration ratio is in favour of the mother, the probability of survival of the fetus is greater than in the opposite situation.

Objective: To search for possible effects of two polymorphisms and of the solar cycle of illumination on reproductive success., Design: Study of haptoglobin (Hp) and ACP1 polymorphisms in consecutive puerperae and analysis of phenotype distribution in relation to time of conception., Setting: The Maternity Department of Penne Hospital, Penne, Italy., Patient(s): Three hundred sixty-eight consecutive healthy pueperae from the Caucasian population., Result(s): The distribution of Hp and ACP1 phenotypes depends on the phase of the solar cycle at conception. Women homozygous for Hp with low ACP1 activity are more likely to conceive in the first part of the year. Women heterozygous for Hp with medium-high ACP1 activity are more likely to conceive in the last part of the year., Conclusion(s): In the first months of the year there is a steady increase in solar illumination, and this phase corresponds to the best period for reproduction in most plants and animals. This period is also the coldest in the Italian latitudes. Although humans are not seasonal breeders, it is possible that women having a genetic background best adapted to the metabolic demand of the cold period of the year will respond better to reproductive stimuli, resulting in a higher probability of conceiving in the first part of the year.

Objectives: To perform a retrospective analysis to assess the utility of pretreatment serum prostatic acid phosphatase (PAP) as a predictor of cause-specific survival (CSS) in patients with higher risk prostate cancer treated with palladium-103 (103Pd) brachytherapy and supplemental external beam radiotherapy (EBRT)., Methods: From 1992 to 1996, 193 patients with clinically localized prostate adenocarcinoma, a pretreatment PAP level, and Gleason score 7 or more, and/or a prostate-specific antigen (PSA) level of 10 ng/mL or more were treated with 103Pd brachytherapy and supplemental EBRT. The patients underwent EBRT of 41.4 Gy to a limited pelvic field and 103Pd brachytherapy with a prescribed minimum 103Pd dose of 80 Gy. Multivariate analysis was performed to analyze the predictive value of PAP, PSA, and Gleason score on CSS., Results: The 10-year CSS rate for patients with a PAP level of less than 1.5, 1.5 to 2.4, and 2.5 U/L or more was 93%, 87%, and 75%, respectively (P = 0.013). The 10-year CSS rate for patients with a PSA level of less than 10, 10 to 20, and greater than 20 ng/mL was 92%, 76%, and 83%, respectively (P = 0.393). The 10-year CSS rate for patients with a Gleason score of 6, 7, 8, and 9 was 90%, 89%, 70%, and 68%, respectively (P = 0.002). On Cox multivariate regression analysis, PAP (hazard ratio 1.31, P <0.0001) and Gleason score (hazard ratio 2.37, P = 0.0007) were associated with CSS. PSA was not predictive of CSS (P = 0.393)., Conclusions: The results of this study demonstrated that PAP is a stronger predictor of CSS than PSA or Gleason score in men with higher risk prostate cancer treated with 103Pd brachytherapy and EBRT. Given the findings of this analysis, the use of PAP should be reconsidered in these patients.

Objectives: Novel prognostic indexes clinically applicable for patients with Stage IV prostate cancer are needed because prostate-specific antigen (PSA) tests occasionally fail to reflect the prognostic outcome. We investigated various clinicopathologic parameters in men with Stage IV prostate cancer and evaluated the utility of the PSA/prostatic acid phosphatase (PAP) ratio as a prognostic index., Methods: We reviewed 241 patients with Stage IV prostate cancer, who were treated in Niigata Cancer Center Hospital from 1992 to 2004. Survival curves were generated using the Kaplan-Meier method. Univariate and multivariate analyses of survival associations, including age, performance status, clinical presentation, disease localization, pathologic findings, and serologic markers, were conducted using the log-rank test and Cox proportional hazard models., Results: The 5-year overall survival rate using the Kaplan-Meier method for all 241 patients was 43.0%. No significant difference was found in the survival rates according to PSA level. However, the 5-year survival rate was significantly lower in patients with a PSA/PAP ratio of less than 3.0 (P = 0.0022): 24.2% and 48.0% in those with a PSA/PAP ratio of less than 3.0 and 3.0 or greater, respectively. On multivariate analysis using the proportional hazards model, the statistically significant prognostic factors of overall survival were alkaline phosphatase (P = 0.0413), lactate dehydrogenase (P = 0.0409), and the PSA/PAP ratio (P = 0.0113)., Conclusions: The PSA/PAP ratio is a valuable prognostic indicator in men with Stage IV prostate cancer. Although our study found that other laboratory tests also had a prognostic influence, the PSA/PAP ratio was an essential index implicated in the physiopathology of prostate cancer.

Objective: To report the results of a trial comparing the efficacy of triptorelin and surgical castration in the treatment of locally advanced or metastatic prostate cancer., Materials and Methods: 80 patients with previously untreated locally advanced or metastatic prostate cancer prostate cancer were included in a one-year multicentre, randomized, prospective, open-label therapeutic trial. Patients either received a monthly injection of triptorelin (group 1; n = 40), or were treated by pulpectomy (group 2; n = 40). Patients were reviewed every 3 months, then every 6 months., Results: The mean age of the patients was 71.22 +/- 8.25 years. At 1 month, 38 patients were castrated (plasma testosterone < 0.5 mg/ml) in the pulpectomy group versus 35 in the triptorelin group. The mean follow-up was 38.8 +/- 26 months in the triptorelin group and 36.3 +/- 25 months in the pulpectomy group. On multivariate analysis, age, impaired performance status and PAP level (> 3.2 ng/ml) were predictive factors of a poor outcome. The median survival was 37.5 +/- 9 months in the triptorelin group and 33 +/- 3 months in the pulpectomy group. At 3 years, no significant difference in specific survival was observed between the 2 groups. At 8 years of follow-up, 63 patients had died., Conclusion: This study demonstrates an equivalent specific survival between patients treated by triptorelin or surgical castration. Castration is rapidly obtained with triptorelin (< 2 months) and is maintained over time throughout the duration of treatment.

In type 1 diabetes mellitus (T1DM), the frequency of antibodies against insulin (IAA), glutamic acid decarboxylase-65 (GAD65), ICA512/IA2 (IA2), and islet cell antigens (ICA) vary with human leukocyte antigen (HLA) composition of the patient. IAA, IA2 autoantibodies, and ICA are increased in DQ8 positives; GAD65 antibodies are increased in DQ2 positives. MHC class I chain-related gene-A (MICA) is another genetic marker that has been proposed to be associated with T1DM. In this article, we looked at microsatellite polymorphism of MICA and its association with autoantibodies (IAA, IA2, and GAD65) in Swedish T1DM patients and if the association explains its importance in early events in autoimmune response. We studied 635 T1DM patients between 0-35 years. Frequency of MICA5/5 was positively associated with the formation of IAA and IA2 antibodies considered individually or in combination (odds ratio [OR], 95% CI, Pc: [IAA+ versus IAA-]: 4.94, 2.09-11.62, <0.0005; [IA2+ versus IA2-]: 2.65, 1.52-4.59, 0.0015; [IAA and/or IA2+ versus rest]: 9.83, 2.37-40.78, <0.0015; [IAA and IA2+ versus rest]: 3.51, 2.01-6.15, <0.0015). Also, -5.1/5.1 was increased in IAA+ patients compared to IAA- patients (2.82, 1.64-4.83, <0.0005). All patients positive for -5/5 developed at least one of the three antibodies. Frequency of MICA5.1 was decreased in IAA+ (0.54, 0.36-0.81, 0.017), in IA2A+ (0.63, 0.45-0.88, 0.04), in IAA and/or IA2A+ (0.52, 0.33-0.84, 0.044), and in IAA and IA2A+ (0.55, 0.39-0.78, 0.0055) patients when compared with patients negative for corresponding antibodies. Frequency of MICA9, 5/5.1, and 5.1/9 was decreased in IAA+ compared to IAA- patients (0.51, 0.32-0.79, 0.021; 0.22, 0.11-0.44, <0.005; and 0.39, 0.22-0.69, 0.026, respectively). Frequency of MICA9 and -5.1/9 was also decreased in IAA and/or IA2 antibody-positive patients while MICA5/5.1 decreased in patients positive for IAA and IA2 antibody both together. IAA and IA2 antibodies are believed to appear early during the autoimmune reaction against beta cells. Thus, according to our data, MICA-5/5 and -5.1/5.1 is associated with early autoimmunity in T1DM patients. Our study suggests that MICA gene polymorphism is associated with autoantibody formation and that the polymorphism especially MICA5/5 and -5.1/5.1 are important in early events of autoimmune reaction.

To evaluate the significance of prostatic acid phosphatase (PAP), we analyzed 1,029 prostate cancer patients who were treated at the Niigata Cancer Center. We classified clinically localized prostate cancer with elevated PAP as stage DO. When stage DO was not taken into acount, the 5-year cause-specific survival rate for stage A, B, C and D was 94.7, 97.9, 87.7 and 42.4%, respectively. Taking stage DO into account, the cause-specific survival curve for stage DO patients was similar to those for stage B,C patients. The 5-year cause-specific survival rate for stage DO was 92.2% considering above 3 ng/ml as elevated PAP. A significant correlation was found between PAP and cause-specific survival for all cases but no correlation was found for non-metastatic disease patients. The significance of PAP in the staging of prostate cancer is limited.

Objective: The "accelerator hypothesis" predicts that fatness is associated with an earlier age at onset of type 1 diabetes. We tested the hypothesis using data from the SEARCH for Diabetes in Youth study., Research Design and Methods: Subjects were 449 youth aged <20 years at diagnosis who had positive results for diabetes antibodies measured 3-12 months after diagnosis (mean 7.6 months). The relationships between age at diagnosis and fatness were examined using BMI as measured at the SEARCH visit and reported birth weight, both expressed as SD scores (SDSs)., Results: Univariately, BMI SDS was not related to age at diagnosis. In multiple linear regression, adjusted for potential confounders, a significant interaction was found between BMI SDS and fasting C-peptide (FCP) on onset age (P < 0.0001). This interaction remained unchanged after additionally controlling for number and titers of diabetes antibodies. An inverse association between BMI and age at diagnosis was present only among subjects with FCP levels below the median (<0.5 ng/ml) (regression coefficient -7.9, P = 0.003). A decrease of 1 SDS in birth weight (639 g) was also associated with an approximately 5-month earlier age at diagnosis (P = 0.008), independent of sex, race/ethnicity, current BMI, FCP, and number of diabetes antibodies., Conclusions: Increasing BMI is associated with younger age at diagnosis of type 1 diabetes only among those U.S. youth with reduced beta-cell function. The intrauterine environment may also be an important determinant of age at onset of type 1 diabetes.

Introduction: We have studied the relationship between the ratio of activated platelets and the thickness of intima and media of the carotid artery in ischemic CVD patients in the chronic stage., Methods: Platelet activation was assessed by means of flow cytometry of whole blood using activation-dependent monoclonal antibodies (MoAb). Forty-one MRI-proven normative subjects and 55 patients with a history of ischemic CVD were examined. The intima-media thickness of the carotid artery was measured by using B-mode ultrasound in all subjects., Results: The appearance rates of PAC-1-positive and CD62P-positive platelets (%) were increased in ischemic CVD patients compared with those in controls (p<0.0001, p<0.001, respectively) The patients and controls were divided into those with atherosclerosis (Ath+), defined as intima-media thickness 1.1 mm, and those without (Ath-). There was no significant difference of PAC-1-positive platelets between the Ath- and Ath+ subgroups in either group, but there was increase in Ath- ischemic CVD patients versus Ath- control subjects (p<0.01), and in Ath+ patients versus Ath+ controls (p<0.05). CD62-positive platelets in the Ath+ subgroup were significantly increased versus the Ath- subgroup in both the controls (p<0.001) and ischemic CVD patients (p<0.05), and there was also an increase in Ath- patients versus Ath- controls (p<0.05)., Conclusion: Platelet activation markers were increased in patients with ischemic CVD compared with controls. A significant relationship was found between increased CD62-P-positive platelets and carotid artery abnormalities in both controls and ischemic CVD patients, suggesting that platelet activation may be a potential marker for atherosclerosis.

In animal models autoreactive CD8(+) T cells are crucial in the development of type 1 diabetes (T1D); however, their role in human T1D is still not known. To address the role of CD81 T cells we performed a pilot study by investigating CD8(+) T cell-mediated cytokine secretion after in vitro stimulation with 94 preproinsulin (PPI) peptides. We were able to show that CD8(+) T cells contribute to a strong IFNgamma reactivity against PPI in human T1D. Further investigations defining epitope specificity, cytokine secretion, and cytotoxic capacity are important to clarify their role in T1D development.

Intravascular large B-cell lymphoma (LBCL) is a rare and aggressive subtype of diffuse LBCL characterized by disseminated intravascular proliferation of neoplastic lymphocytes. Obstruction of blood flow by tumor cells in a variety of organs can cause an array of clinical changes, including alteration of the neural and spinal system and the respiratory system, as well as skin lesions. It is usually very difficult to diagnose intravascular LBCL in a patient simply from clinical symptoms or laboratory examinations. We here document our findings that serum prostatic acid phosphatase levels in both males and a female (2.2-24.0 microg/L) reflect the presence of intravascular LBCL, changing synchronously in response to chemotherapy. To determine whether prostatic acid phosphatase (PAP) might be a useful tumor marker for early diagnosis, we reviewed five intravascular LBCLs. Immunohistochemically, tumor cells in all cases were positive for anti-PAP antibody. The results were further confirmed in one case by Western-blot analysis and in another by the detection of amplified messenger RNA for PAP in microdissected tumor cells, respectively. PAP has not been detected in 17 lymphomas (diffuse LBCL, 8 cases; follicular lymphoma, 3 cases; T-cell lymphoma, 3 cases; Hodgkin lymphoma, 3 cases) by Western blot analyses. We conclude that serum PAP is a useful tumor marker for intravascular LBCL and that it deserves further investigation in this context.

Lysophosphatidic acid (LPA) is a lipid mediator with multiple biological activities and is detected in various biological fluids, including human seminal plasma. Due to its cell proliferation stimulatory and anti-apoptotic activities, LPA has been implicated in the progression of some cancers such as ovarian cancer and prostate cancer. Here, we show that prostatic acid phosphatase, which is a non-specific phosphatase and which has been implicated in the progression of prostate cancer, inactivates LPA in human seminal plasma. Human seminal plasma contains both an LPA-synthetic enzyme, lysoPLD, which converts lysophospholipids to LPA and is responsible for LPA production in serum, and its major substrate, lysophosphatidylcholine. In serum, LPA accumulated during incubation at 37 degrees C. However, in seminal plasma, LPA did not accumulate. This discrepancy is explained by the presence of a strong LPA-degrading activity. Incubation of LPA with seminal plasma resulted in the disappearance of LPA and an accompanying accumulation of monoglyceride showing that LPA is degraded by phosphatase activity present in the seminal plasma. When seminal plasma was incubated in the presence of a phosphatase inhibitor, sodium orthovanadate, LPA accumulated, indicating that LPA is produced and degraded in the fluid. Biochemical characterization of the LPA-phosphatase activity identified two phosphatase activities in human seminal plasma. By Western blotting analysis in combination with several column chromatographies, the major activity was revealed to be identical to prostatic acid phosphatase. The present study demonstrates active LPA metabolism in seminal plasma and indicates the possible role of LPA signaling in male sexual organs including prostate cancer.

Background and Objective: Intravascular large B-cell lymphoma (IVL) has been treated as fever of unknown origin (FUO), and many patients have been treated inadequately based on incorrect diagnoses. We previously cares for a patient with IVL who tested positive for prostatic acid phosphatase (PAP), a marker of prostate cancer. Since then, we have regularly examined this mather when IVL was suspected to investigate the usefulness of PAP as a diagnostic marker for IVL. We retrospectively evaluated the usefulness of PAP as diagnostic marker of IVL., Design and Methods: We reviewed the clinical courses of 5 patients with IVL (3 males, 2 females) in comparison with 23 controls with hematologic malignancies other than IVL., Results: Serum levels of PAP were elevated in all 5 patients with IVL and 2 of the 23 controls. The difference was statistically significant using a chi-squared test (p=0.0002). The sensitivity and specificity of PAP were 100% and 91%, respectively, in the diagnosis of IVL. Its serum levels were closely associated with disease status., Interpretation and Conclusions: This study suggests that PAP might be a useful marker for the screening and assessment of disease activity and responses to the treatment of IVL.

Objectives: To explore the relationships between the peripheral blood levels of CD61, CD63, PAC-1 and the incidence of acute rejection and tubular necrosis after renal transplantation, and recovery of the graft function., Methods: The peripheral blood levels of CD61, CD63, and PAC-1 of 86 patients with uremia in different stages before and after transplantations were analyzed by flow cytometry. The patients were divided into three groups: (1) twenty-nine patients with normal grafts function, (2) hirty with acute rejection and (3) twenty-seven with acute tubular necrosis. The patients with acute rejection were randomly divided into treatment group with anticoagulants and cntrol group., Results: The peripheral blood levels of CD61, CD63 and PAC-1 significantly increased (P < 0.05) in the patients with acute rejection, in comparison with those with normal grafts function and those with acute tubular necrosis. The peripheral blood levels of CD61, CD63 and PAC-1 in patients with acute rejection in anticoagulants therapy was lower, recovery time of the grafts function was shorter, one-year survival rates of patients and grafts were higher, as compared with those of controls., Conclusions: The patients with acute rejection have significantly high peripheral blood levels of CD61, CD63 and PAC-1 before transplantation, however, these values in patients with acute tubular necrosis are not high, this suggesting that acute rejection might relate to platelet activation, while acute tubular necrosis might not relate to it. After anticoagulants therapy in patients with acute rejection, the grafts function might recover faster and their one-year survival rates and grafts might be higher in those with CD61, CD63 and PAC-1 decreasing remarkably.

Background: New cases of type 1 diabetes mellitus (DM1) in Slovak children accumulate in late summer, autumn and winter., Hypotheses: Children manifesting the disease in these seasons have higher autoantibody positivity than those first diagnosed in spring or summer., Patients and Methods: One hundred and fifty Slovak children (and adolescents), aged 1-14 (15-17) years at the manifestation of DM1, born 1978-2000, with the disease manifested 1989-2001, were investigated at diagnosis by IA-2A, GADA and IAA autoantibody positivity, using standard radioimmunoassay procedures. The resulting risk score values (0-1) were related to calendar days of diagnosis. Their annual, semi-annual and quarterly periodicity was tested using Halberg's cosinor regression., Results: Only IA-2A positivity at the time of diagnosis showed significant seasonal cycling, with the acme around the autumn equinox and with the nadir in spring. There was no increase of this positivity in winter., Conclusion: The maximum autoantibody levels were found for those manifesting the disease in late summer and early autumn--the only time of year in Slovakia when both the numbers of births of future diabetic children as well as those of manifestation of new cases are significantly above average.

Content of different cytokines (IF alpha, TNF alpha, IL-1 alpha, IL-4, IL-6 and IL-10) was examined in the blood serum in two groups of healthy children-siblings with type 1 diabetes mellitus with and without revealed insulin autoantibodies against pancreatic islets (GADA, IA-2A and IAA) by enzyme-like immunosorbent assay (ELISA). In the group of patients with two and more revealed autoantibodies the higher indices in the number of IF alpha, TNF alpha and IL-6, and the decrease in the level of IL-4 comparing with the group of children with negative reaction to diabetes associated autoantibodies were more often observed.

Background: In animal models, heparin coating reduces platelet accumulation induced by coronary stenting. However, reduced platelet activation has never been demonstrated in humans. The purpose of the current study was to investigate the effect of heparin coating on platelet activation after coronary artery stenting., Methods: In a prospective, randomized, pilot study of 50 consecutive elective patients, platelet activation was analyzed by measuring aggregation and surface receptor expression before and at 2 hours, 24 hours, 5 days and 30 days after implantation of either heparin-coated or noncoated stents., Results: There was less platelet activation after implantation of a heparin-coated stent, as indicated by reduced expression at 24 hours of active glycoprotein (GP) IIb/IIIa (10.3 +/- 6.1 vs 6.7 +/- 2.1, P =.014), and total GP IIb/IIIa (382 +/- 101 vs 306 +/- 88, P =.01). A trend at 30 days poststenting of lower total (383 +/- 150 vs 296 +/- 86, P =.07) and active GP IIb/IIIa expression (10.3 +/- 6.9 vs 7.5 +/- 2.9, P =.15) was also observed with the heparin-coated stent). Aggregation and stimulated p-selectin did not differ between stent types., Conclusion: Use of a heparin-coated stent in this pilot study of elective patients was associated with primarily less early platelet expression of GP IIb/IIIa. These findings have implications on the risk of subacute thrombosis and deserve further investigation.

Objective: To find out Gleason grades, scores and to see the correlation of these morphological features with tumour markers in prostatic carcinoma., Design: A descriptive study., Place and Duration of Study: The study was conducted at the Departments of Histopathology and Chemical Pathology, Armed Forces Institute of Pathology, Rawalpindi, over a period of one year., Materials and Methods: Fifty cases of prostatic carcinoma were studied. Gleason grades and score of tumour were determined by doing haematoxylin and eosin (HE) staining. Pre-operative serum prostatic acid phosphatase (PAP) and prostate-specific antigen (PSA) assays were carried out in these cases., Results: The patients seen were between 50-102 years of age with an average of 70.9 years. There were 49 cases of adenocarcinoma and 01 case of mixed adeno and transitional cell carcinoma of prostate. Twenty-eight (56%) patients had Gleason score of 5-7. Twenty-nine (58%) patients were having serum PSA levels between 10.0 ng/ml and 50.0 ng/ml. Thirteen (26%) cases showed PSA assays >50 ng/ml. The sensitivity of PSA test was 84 % in these cases. Thirty-five (70%) patients were having PAP values >3.7 U/l (sensitivity 70 %)., Conclusion: The Gleason grading system is a specific morphological predictor. The serum PSA showed better sensitivity and specificity with Gleason grades and scores as compared to serum PAP. The serum PAP levels showed better correlation with morphological features as compared to serum PSA.

Objective: To analyze the relationships between the expression levels of CD61, CD63, and PAC-1 on the platelet surface and the incidences of acute rejection and tubular necrosis as well as the recovery of graft function after renal transplantation., Methods: The expression levels of CD61, CD63, and PAC-1 on platelet surfaces were assayed by flow cytometry in 86 patients with different stages of uremia before and after transplantation. Patients were divided into three groups: 29 patients with normal graft function, 30 with acute rejection, and 27 with acute tubular necrosis. Patients with acute rejection were randomly assigned into groups treated with or without anticoagulants., Results: The expression levels of CD61, CD63, and PAC-1 on platelet surfaces significantly increased (P <.05) among patients with acute rejection, as compared with those with normal graft function or acute tubular necrosis. Compared with controls, the expression levels of CD61, CD63, and PAC-1 were lower among acute rejection patients who, received anticoagulant therapy. The recovery time for graft function shorter and, the 1-year patients and graft survival rates higher., Conclusions: The pretransplant expression levels of CD61, CD63, and PAC-1 on platelet surface were significantly higher among patients with acute rejection, suggesting that this complication rather than acute tubular necrosis may be related to platelet activation. Patients with acute rejection displayed benefit from anticoagulant therapy.

Islet cell autoantibodies are early markers for type 1 diabetes. The aim of this study was to determine whether islet autoantibodies were present at birth in young adults who developed type 1 diabetes at 15-30 years of age. Cord blood sera from 30 patients who developed type 1 diabetes between 15 and 25 years of age and sera from 320 randomly selected control children were tested for islet cell antibodies (ICA) by indirect immunofluorescence and autoantibodies against the 65 kD isoform of glutamic acid decarboxylase (GADA), islet cell antigen-2 (IA-2A) and insulin (IAA) by radiobinding assays. The young adults who developed type 1 diabetes did not differ from controls in the cord blood prevalence of any of the four islet autoantibodies. This is in contrast to our previous findings that children who developed type 1 diabetes below 15 years of age had an increased prevalence of cord blood islet autoantibodies. Our present data suggest that, in contrast to children, pre- and perinatal risk factors are less likely to be involved in the development of type 1 diabetes in young adults.

PSA continues to be one of the most effective and widely used cancer screening tools available. Its popularity in prostate cancer screening, however, has eroded its usefulness in the staging of this disease. As more men are screened every year on a routine basis with DRE and PSA, the average PSA at diagnosis has drifted down to well below 10 ng/mL in many centers, including ours. This trend is likely to accelerate, as a PSA cut off for prompting biopsy of the prostate of 2.5 ng/mL gains more widespread acceptance. The recent realization that, at these levels, serum PSA is more reflective of the presence of BPH than of the extent of cancer and, therefore, does not provide additional staging information, has renewed the search for new biochemical markers that are capable of predicting prostate cancer stage and prognosis. Because of the heterogeneity of this disease, it is unlikely that a single biochemical marker that is capable of accurately staging all prostate cancer patients will be found. For this reason, nomograms that are capable of integrating various parameters to predict stage and prognosis will remain indispensable. As new biochemical markers that provide independent predictive information about stage or prognosis are identified, they can be incorporated into currently available nomograms. Of the biochemical markers discussed in this article, IL-6sR and TGF-beta1 are the most promising. By incorporating them into a preoperative nomogram designed to predict PSA recurrence, we found that they improved the ability to predict biochemical recurrence by a statistically and clinically significant margin. The ability to stage prostate cancer and predict response to therapy has improved dramatically over the last 3 decades. Nevertheless, there is still a need for new biochemical markers that will improve the ability to predict an individual patient's stage and response to therapy. Incorporating these new markers into nomograms will enhance the ability to provide optimal care for each prostate cancer patient.

Aims: The autoantigen SOX-13 of the SRY-related high mobility group box is a low-frequency reactant in sera from patients with Type 1 diabetes. We further investigated the potential diagnostic role of anti-SOX-13, and in particular its ability to distinguish Type 1 from Type 2 diabetes, in two large, well-characterized cohorts., Methods: SOX-13 autoantibody status was ascertained using a radioimmunoprecipitation assay in (i) a random sample of 546 participants in an Australian community-based study (the Fremantle Diabetes Study; FDS) of whom 119 had Type 1 and 427 Type 2 diabetes, and (ii) a sample of 333 subjects with Type 2 diabetes from the United Kingdom Prospective Diabetes Study (UKPDS) stratified by age, anti-glutamic acid decarboxylase (GAD) and islet cell antibody (ICA) status, and requirement for insulin therapy within 6 years of diagnosis., Results: The frequencies of anti-SOX-13 in the FDS subjects were 16.0% and 14.8% for Type 1 and Type 2 patients, respectively, and levels were similar. In the UKPDS subjects, the frequency was 4.5%. In a logistic regression model involving demographic, anthropometric and metabolic variables, only diabetes duration was significantly associated with anti-SOX-13 positivity, especially for duration > 5 years (P < 0.002). When the coexistence of autoantibodies was assessed in the two study samples, there were no significant associations between anti-SOX-13 and ICA, anti-GAD or ICA512/IA-2., Conclusions: Whilst the frequency of anti-SOX-13 may be increased in some populations of diabetic patients, this reactivity does not usefully distinguish Type 1 from Type 2 diabetes. However, the association with diabetes duration suggests that anti-SOX-13 may be a non-specific marker of tissue damage associated with chronic hyperglycaemia.

Background: ACP1, also called cLMWPTP (cytosolic Low Molecular Weight PTPase) is a highly polymorphic enzyme involved in the modulation of signal transduction by insulin, PDGF receptors, and T-cell receptors. The enzyme is controlled by a locus on chromosome 2, with three common codominant alleles; the corresponding six genotypes show strong variations in total enzymatic activity. The purpose of our research was to determine the relationship of ACP1 with glycemic level, ketoacidosis and HbA1C in children with Type 1 diabetes., Material/methods: We studied 189 consecutive children with Type 1 diabetes, from the Pediatric Clinic of Sassari University. The ACP1 genotype was determined by PCR and digestion by specific restriction enzymes., Results: At initial diagnosis a strong negative correlation was observed between glycemic level and ACP1 activity, with the highest levels in genotypes with low activity. Ketoacidosis and HbA1C show a similar pattern of relationship with ACP1. A comparative analysis of the data on Type 1 diabetes with previously obtained data on Type 2 diabetes shows an opposite pattern of relationship between ACP1 and metabolic parameters. Moreover, correlation between glycemia and HbA1C in Type 1 diabetes is much weaker than in Type 2 diabetes., Conclusions: These differences suggest that the enzyme might be involved in different signal transduction pathways relevant in the pathogenesis of these two classes of diabetic disorders. It would be interesting to study the possible correlation in Type 1 diabetes between ACP1 and immunological parameters.

Objectives: patients with peripheral arterial disease (PAD) have a threefold increase in cardiovascular mortality. Standard antiplatelet treatment may not confer uniform benefit in different patient groups. This study aimed to compare platelet function in patients with lower limb PAD, carotid disease and abdominal aortic aneurysm (AAA) with age- and sex-matched healthy controls., Methods: patients with lower limb PAD (n = 20), carotid disease (n = 40), AAA (n = 13) and age/sex matched healthy controls (n= 20) were studied. Whole blood methods to detect spontaneous platelet aggregation (SPA), and adenosine diphosphate (ADP) and collagen-induced aggregation were used. The detection of platelet P-selectin and the PAC-1 antigen by flow cytometry were also used as markers of platelet activation and aggregation., Results: patients with lower limb PAD or AAA had higher baseline SPA compared to normal controls (p < 0.01). There was significantly higher collagen-induced aggregation in IC patients compared to normal controls (p < 0.01). However, there was no difference in ADP-induced aggregation between lower limb PAD and control patients. There was no difference in PAC-1 binding between control patients and the patients with lower limb PAD, carotid disease or AAA. Patients with carotid disease had a higher expression of P-selectin compared to normal controls (p < 0.05)., Conclusions: this study provides further evidence that platelet hyperactivity is present in patients with PAD despite the use of antiplatelet therapy. Further antiplatelet strategies may be indicated to protect these patients.

Background: The continuous measurement of acid phosphatase (EC 3.1.3.2) activity in serum represents an analytical task not yet sufficiently accomplished., Methods: Introducing two novel substrates-2-chloro-4-nitrophenyl phosphate (CNP-P), which was preferred, and 4-nitronaphthyl-1-phosphate (NN-P)-an alternative assay to measure enzymatic activity was developed and compared with a modification of Hillmann's method (azo coupling of released naphth-1-ol with a diazonium compound). Apart from different substrate concentrations of 2-chloro-4-nitrophenyl phosphate, 4 mmol/l, and naphthyl-1-phosphate (N-P), 8 mmol/l (with Fast Red TR, 5 mmol/l), respectively, following identical conditions were selected: Citrate, 50 mmol/l, pH 5.75; pentane-1,5-diol, 150 mmol/l; tartrate, 60 mmol/l; 37 degrees C., Results: Whereas intensity and stability of the azo dye unpredictably depend on the albumin concentration of the sample, the direct test with 2-chloro-4-nitrophenyl phosphate resisted sample interferences, showed no intrinsic hydrolysis by albumin, relied on stable reagents and proved superior in sensitivity, precision and ease of handling. In measuring prostatic phosphatase, the proposed procedure closely correlated with Hillmann's method. The preliminary 0.95-reference intervals for adults were 1.2-3.9 kU/l and 5.8-14.8 U/l for total activity, respectively., Conclusions: The direct assay of the enzyme is suited as an economic, rapid and robust method for mechanized or manual use.

The role of calpain in platelet function is generally associated with aggregation and clot retraction. In this report, data are presented to show that one component of the platelet secretory machinery, SNAP-23, is specifically cleaved by calpain in activated cells. Other proteins of the membrane fusion machinery, e.g. syntaxins 2 and 4 and alpha-SNAP, are not affected. In vitro studies, using permeabilized platelets, demonstrate that cleavage is time- and calcium-dependent. Analysis of SNAP-23 cleavage products suggests that the calpain cleavage site(s) is in the C-terminal third of the molecule potentially between the cysteine-rich acyl attachment sites and the C-terminal coiled-coil domain. The time course of cleavage is most consistent with late calpain-mediated events such as pp60(c-src) cleavage, but not early events such as protein-tyrosine phosphatase-1B activation. SNAP-23 cleavage is inhibited by calpeptin, calpastatin, calpain inhibitor IV, and E-64d, but not by caspase 3 inhibitor III or cathepsin inhibitor I. When tested for their effect on secretion, none of the calpain-specific inhibitors significantly affected release of soluble components from any of the three platelet granule storage pools. These results indicate that SNAP-23 cleavage occurs after granule release and therefore may play a role in affecting granule membrane exteriorization. This is consistent with the ultrastructural morphology of calpeptin-treated platelets after activation.

This was a ten-year, hospital-based retrospective study for the incidence and clinical pattern of prostate cancer in southeastern Nigeria. Clinical information extracted from the files included the TNM stage, histo-pathological grading, level of prostatic acid phosphatase (PAP), mode of presentation and clinical and biochemical response to intravenous and oral diethylstilboestrol diphosphate (Honvan)/ orchidectomy. There were 145 patients, mean age 66.6 + 9.8 years, giving an incidence of 61.3 per 10(5), with 54% under 70 years. Most patients (81.4%) presented late, with 62% metastatic. Over 98% were adenocarcinomas, 77% of which were moderate to well-differentiated cancers. PAP was elevated in 109 patients (75%), (representing 92% of all advanced tumours), and normal in 36 (25%). Forty-two percent of poorly differentiated cancers had normal levels of PAP. Most patients presented with urinary retention (56%), prostatism (44%), anaemia (41%), recurrent UTI (35%), bone pains (20%), haematuria (18%), backache (16%) and paraplegia (6%). Nearly 79% responded to treatment with lowered PAP levels and improved quality of life, within a mean of 26.3+/-13.8 months (range 5-78); objective 81 (58%), subjective 32 (23%), no response 27 (19%). Among paraplegics, 78% had full, and 22% had partial motor recovery. Patients with poorly differentiated cancers had only a 33% two-year survival rate. This study confirmed an upward, though moderate trend in the incidence of prostate cancer in Nigeria. The use of PAP instead of PSA as the tumor marker, a local diet with high fish content but lower animal fat, and poor hospital access may account for the lower incidence in the southeast. Poor health education may account for the high rate of late presentations.

Background: The level of prostatic acid phosphatase in serum is an established marker for prostate carcinoma., Methods: Inactivation of homogeneous prostatic acid phosphatase from human seminal fluid by purified plasmin and human serum was studied in the presence and absence of bovine pancreatic trypsin inhibitor, a plasmin inhibitor, or phenylmethylsulfonylfluoride, a serine protease inhibitor., Results: Plasmin or serine protease inhibitors protect against prostatic acid phosphatase inactivation in serum samples., Conclusion: The immediate addition of serine protease inhibitors to serum samples taken for prostatic acid phosphatase determinations should provide more accurate results and permit extended storage of samples. The stabilization of the enzyme activity and immunological properties of prostatic acid phosphatase in blood samples by these protease inhibitors resurrects the clinical significance of prostatic acid phosphatase measurements in prostate cancer screenings.

The incidences of cardiovascular and cerebrovascular diseases are reportedly higher in patients with obstructive sleep apnea (OSA) than in OSA-free subjects, though the mechanism remains unknown. Recently, the contribution of activated platelets to a number of pathological conditions such as stroke or ischemic heart disease has been suggested. We hypothesized that the expression of activated platelet markers resulting from OSA might be higher than in healthy subjects. By flow cytometry using monoclonal antibodies, we measured two such markers, PAC-1 and CD 62 P, in OSA patients and healthy subjects. Twelve healthy men (age, 52.7 +/- 12.8 y/o; and body mass index (BMI), 22.2 +/- 16.1 kg/m2; mean +/- S.D.) and 20 male patients with OSA (age, 50 +/- 7.96 y/o; BMI, 28.1 +/- 3.3 kg/m2; apnea hypopnea index (AHI), 38.2 +/- 21.2 times/hr; and lowest SpO2, 75.6 +/- 11.3%) were enrolled in this study. PAC-1 expression was significantly higher in OSA patients (65.1 +/- 17.8%) than in healthy subjects (16.8 +/- 7.4%), as was CD 62 P expression (8.5 +/- 8.8% vs. 0.88 +/- 0.57%). The increase in PAC-1 expression was correlated with AHI and the arousal index. These findings suggest that activated platelet markers could be good indicators for untreated OSA.

Platelets were activated with freezing/thawing and thrombin stimulation, and platelet microparticles generated following platelet activation were isolated with ultracentrifugation. The effects of platelet microparticles on platelet activation were studied with annexin V assay, protein tyrosine phosphorylation, and platelet aggregation. Freezing-induced platelet microparticles decreased but thrombin-induced platelet microparticles increased platelet annexin V binding and aggregation. Freshly washed platelets were cryopreserved using epinephrine and dimethyl sulfoxide (Me(2)SO) as combined cryoprotectants, and stimulated with thrombin-induced platelet microparticles. Following incubation of thrombin-induced platelet microparticles, the reaction time of platelets to agonists decreased but the percentages of aggregation increased, such as washed platelets from 44% +/- 30 to 92% +/- 7, p < 0.001, and cryopreserved platelets from 66% +/- 10 to 77% +/- 7, p < 0.02. By increasing platelet aggregability, platelet microparticles recovered after thrombin stimulation improved platelet function for transfusion. A 53-kDa platelet microparticle protein showed little phosphorylation if it was released from resting platelets or platelets stimulated with ADP, epinephrine, propyl gallate or dephosphorylation if it was derived from ionophore A 23187-stimulated platelets. However, the same protein released from frozen platelets showed significant tyrosine phosphorylation. Since a microparticle protein with 53 kDa was compatible with protein tyrosine phosphatase-1B (PTP-1B), its phosphorylation suggests the inhibition of enzyme activity. The microparticle proteins derived from thrombin-stimulated platelets were significantly phosphorylated at 64 kDa and pp60c-src, suggesting that the activation of tyrosine kinases represents a possible mechanism of thrombin-induced platelet microparticles to improve platelet aggregation.