New trends in prostate cancer diagnostics.

The ever-increasing incidence of prostate cancer is driving research and clinicians efforts to manage the disease more precisely. Leaving aside the progress we are making in the field of prostate cancer treatment, we can notice the pressure on accurate diagnosis in everyday practice. We often meet patients who are in the so-called gray zone and we are not sure whether to indicate a prostate biopsy or continue to monitor the patient. For diagnostics, we use a number of more or less proven methods that we have at our disposal. No clinical urologist wants to send their patient for a prostate biopsy unnecessarily. However, if the patient is finally planned for a prostate biopsy, there is no 100% certainty that we will detect the cancer. For this reason, there is an effort to develop other methods that would refine the collection of histological material and thus increase the capture of those patients who need treatment. The aim of this review is to show modern approaches to diagnostics and outline the direction in which diagnostics will go in the coming years.

Risk of biochemical recurrence in patients with a positive surgical margin after radical prostatectomy.

INTRODUCTION: Serum prostate specific antigen (PSA) is an irreplaceable marker in the detection and follow-up of patients with prostate cancer. In our analysis we addressed factors that could indicate the likelihood of biochemical recurrence (BCR) early after radical prostatectomy. We mainly focused on the positive surgical margin (R1). METHODS: Retrospective evaluation and analysis of the database of patients with prostate cancer after radical prostatectomy from 2001 to 2019. In total 1529 patients were enrolled in the study. The median follow-up was 48 months. The age of the patients ranged from 49 to 76 years. We used pre-operative PSA values, and the monitoring of the dynamics of 3rd generation PSA progression (detection limit 0.003 ng/ml) at month 1 and month 3 after surgery and then in 3-month intervals. We monitored the surgical margin positivity (R0 negative, R1 positive) and the Gleason score (GS) based on histological samples and we analysed the relationship to biochemical recurrence of the disease. RESULTS: The pre-operative PSA value did not show a direct relationship to the R1 risk. Patient values in the groups R1 and R0 differed only by 1.159 ng/ml (p=NS). The 3rd generation PSA value at month 1 after surgery was 50.82% higher in R1 patients (p>0.001). 50% of patients with R1 (29.5% patients of the total) did develop BCR during the follow-up period, while in patients with R0 (70.5% patients of the total) this proportion was 30% (p>0.001). Among those with GS 67, 47% developed BCR. The GS 810 group relapsed in 75% of the cases (p>0.001). CONCLUSION: According to our analysis 33% of the patients reached the stage of biochemical recurrence. We demonstrated a direct dependency between the risk of recurrence and the final Gleason score. The presence of R1 should not be viewed as a direct indication for adjuvant radiotherapy.

Signalling by the W/Kit receptor tyrosine kinase is negatively regulated in vivo by the protein tyrosine phosphatase Shp1.

Protein tyrosine phosphorylation plays a key role in regulating eukaryotic cell proliferation and differentiation. Genetic analysis in invertebrates has been invaluable for dissecting the signalling events downstream of receptor tyrosine kinases (RTKs). We have used this approach in mammals to analyse the interactions between the Kit RTK encoded by the murine Dominant white spotting (W) locus and the Shp1 protein tyrosine phosphatase, the product of the murine motheaten (me) gene. Homozygosity for mutations in both W and me ameliorates aspects of both the me and W phenotypes, including the lethal lung disease associated with me and the embryonic lethality and mast cell deficiency associated with W, demonstrating that the Kit receptor plays a role in the pathology of the me phenotype and conversely that Shp1 negatively regulates Kit signalling in vivo.

Peripheral blood marker of residual acute leukemia after hematopoietic cell transplantation using multi-plex digital droplet PCR.

Background: Relapse remains the primary cause of death after hematopoietic cell transplantation (HCT) for acute leukemia. The ability to identify minimal/measurable residual disease (MRD) via the blood could identify patients earlier when immunologic interventions may be more successful. We evaluated a new test that could quantify blood tumor mRNA as leukemia MRD surveillance using droplet digital PCR (ddPCR). Methods: The multiplex ddPCR assay was developed using tumor cell lines positive for the tumor associated antigens (TAA: WT1, PRAME, BIRC5), with homeostatic ABL1. On IRB-approved protocols, RNA was isolated from mononuclear cells from acute leukemia patients after HCT (n = 31 subjects; n = 91 specimens) and healthy donors (n = 20). ddPCR simultaneously quantitated mRNA expression of WT1, PRAME, BIRC5, and ABL1 and the TAA/ABL1 blood ratio was measured in patients with and without active leukemia after HCT. Results: Tumor cell lines confirmed quantitation of TAAs. In patients with active acute leukemia after HCT (MRD+ or relapse; n=19), the blood levels of WT1/ABL1, PRAME/ABL1, and BIRC5/ABL1 exceeded healthy donors (p<0.0001, p=0.0286, and p=0.0064 respectively). Active disease status was associated with TAA positivity (1+ TAA vs 0 TAA) with an odds ratio=10.67, (p=0.0070, 95% confidence interval 1.91 - 59.62). The area under the curve is 0.7544. Changes in ddPCR correlated with disease response captured on standard of care tests, accurately denoting positive or negative disease burden in 15/16 (95%). Of patients with MRD+ or relapsed leukemia after HCT, 84% were positive for at least one TAA/ABL1 in the peripheral blood. In summary, we have developed a new method for blood MRD monitoring of leukemia after HCT and present preliminary data that the TAA/ABL1 ratio may may serve as a novel surrogate biomarker for relapse of acute leukemia after HCT.

Soluble form of carbonic anhydrase IX (CAIX) in transitional cell carcinoma of urinary tract.

We investigated the expression of cell-associated CAIX protein in histological sections of the transitional cell carcinoma (TCC) of the urinary tract and of the soluble form of CAIX (s-CAIX) shed by the tumor into the serum and urine of TCC patients. A total of 23 patients with histologically confirmed TCC or squamous cell carcinoma (SCC) were enrolled in the pilot study. Sixteen healthy individuals served as controls. Membrane-bound CAIX was present in the tumor cells near the endoluminal surface. Necrosis was observed in only 4 samples. Using Western blots, s-CAIX concentrated from urine was visualized as a double band at 50 and 54 kDa. In most cases, the presence of s-CAIX in the urine correlated with CAIX expression in the tumor. On the other hand, s-CAIX did not exceed the normal level in the serum of TCC patients. Urine from patients with TCC of the urinary bladder and renal pelvis contained s-CAIX, allowing the detection of tumors in approximately 70% of the patients. Moreover, two additional patients with suspected, but unconfirmed bladder tumor, with s-CAIX detected in urine, developed tumors identified as TCC within six months. We suggest that after a simple, rapid and sensitive test, monitoring s-CAIX levels in urine will be developed, it may be useful for early detection of relapse in patients following transurethral tumor resection.

Dynamics of microvesicle generation in B-cell chronic lymphocytic leukemia: implication in disease progression.

Previously, we reported that B-cell chronic lymphocytic leukemia (CLL) patients contained elevated levels of microvesicles (MVs). However, given the quiescent nature of CLL B-cells and the relative indolence of the disease, the dynamics of MV generation and their unique phenotypes are not clearly defined. In this study, we find that CLL B-cells generate MVs spontaneously and can be further induced by B-cell receptor-ligation. Most interestingly, CLL B-cells predominantly generate CD52+ MVs, but not CD19+ MVs in vitro, suggesting preferential usage of CD52 into leukemic-MVs and that the CLL plasma MV phenotypes corroborate well with the in vitro findings. Importantly, we detected increased accumulation of CD52+ MVs in previously untreated CLL patients with progressive disease. Finally, sequential studies on MVs in pre- and post-therapy CLL patients demonstrate that although the plasma CD52+ MV levels drop significantly after therapy in most and remain at low levels in some patients, a trend of increased accumulation of CD52+ MVs was detected in majority of post-therapy CLL patients (25 of 33). In total, this study emphasizes that dynamic accumulation of CD52+ MVs in plasma can be used to study CLL progression and may be a useful biomarker for patients as they progress and require therapy.

The incidence of thrombotic thrombocytopenic purpura-hemolytic uremic syndrome: all patients, idiopathic patients, and patients with severe ADAMTS-13 deficiency.

BACKGROUND: Accurate estimates of the incidence of thrombotic thrombocytopenic purpura (TTP) are important to assess the resources required for current treatments as well as to anticipate the need to develop new treatments. Previous estimates have been indirect and have not reported data on patients with ADAMTS-13 deficiency. OBJECTIVE: To determine the incidence of patients with TTP-hemolytic uremic syndrome (HUS) in three categories: all patients with clinically suspected TTP-HUS, patients with idiopathic TTP-HUS, and patients with severe ADAMTS-13 deficiency. METHODS: Incidence rates were estimated from the Oklahoma TTP-HUS Registry, analyzing all 206 consecutive patients from January 1, 1996 to June 30, 2004 who were treated with plasma exchange for their initial episode of clinically suspected TTP-HUS. ADAMTS-13 activity was measured in 186 (90%) of the 206 patients. RESULTS: The age-sex-race standardized annual incidence rates were 11.29 x 10(6) (95% CI: 9.70-12.88) for all patients with clinically suspected TTP-HUS; 4.46 x 10(6) (95% CI: 3.43-5.50) for patients with idiopathic TTP-HUS; and 1.74 x 10(6) (95% CI: 1.06-2.41) for patients with severe ADAMTS-13 deficiency (<5% activity). In all three categories, the incidence rates were greater for women and for blacks. For patients with severe ADAMTS-13 deficiency, the age-sex standardized incidence rate ratio of blacks to non-blacks was 9.29 (95% CI: 4.33-19.93). CONCLUSIONS: Accurate incidence rate estimates for all patients with clinically suspected TTP-HUS, idiopathic TTP-HUS, and TTP associated with severe ADAMTS-13 deficiency have been determined. The greater incidence among women and blacks is comparable with their increased risk for other autoimmune disorders.

Life after acquired thrombotic thrombocytopenic purpura: morbidity, mortality, and risks during pregnancy.

Patients who have recovered from their acute episode of acquired ADAMTS13-deficient thrombotic thrombocytopenic purpura (TTP) were once thought to have complete recovery except for risk of relapse. Data from previous publications from the Oklahoma TTP-hemolytic uremic syndrome (HUS) Registry are summarized. Patients have decreased cognitive function and increased prevalence of hypertension, systemic lupus erythematosus, major depression, and albuminuria as compared to the expected values from the US population. The proportion of patients that died during the follow-up period was greater than expected based on the US population reference population. Among women who had a pregnancy following recovery from TTP, relapse during pregnancy or postpartum is uncommon, but the occurrence of preeclampsia may be increased. Thirteen of 16 pregnancies in these women resulted in healthy children. Increased morbidity and mortality in TTP patients following recovery suggest that TTP may be more of a chronic disorder than a disorder with acute episodes and complete recovery.

Management of patients with chronic, refractory idiopathic thrombocytopenic purpura.

Chronic refractory idiopathic thrombocytopenic purpura (ITP) is defined as ITP with persistent thrombocytopenia despite conventional initial management with prednisone and splenectomy. Rare in children, It may occur in as many as one third of adults with ITP. The goal of treatment is not cure of the ITP, but only to achieve a safe platelet count, which is arbitrarily assumed to be greater than 30,000 to 50,000/microL. The risk for major bleeding seems great only when the platelet count is less than 10,000/microL. Treatment of patients with moderate thrombocytopenia and no clinically important bleeding symptoms should be avoided. There is no accepted algorithm for management of patients with chronic refractory ITP. Observation without specific treatment must be considered a cornerstone of management. Combination regimens of Immunosuppressive agents may be required for patients with severe and symptomatic thrombocytopenia. Additional supportive care measures are also important.

Thrombotic thrombocytopenic purpura-like syndromes following bone marrow transplantation: an analysis of associated conditions and clinical outcomes.

The diagnosis and treatment of thrombotic thrombocytopenic purpura (TTP) in patients following BMT are often uncertain and unsuccessful. To better understand the evaluation and management of these patients, we describe 17 patients treated with plasma exchange for a presumptive diagnosis of TTP following BMT during a 10 year period, 1989-1998. Because of the uncertainty of the diagnosis, these patients are described as having a 'TTP-like syndrome'. All 17 patients had received an allogeneic BMT. Comparison with the other 245 patients who had an allogeneic BMT during the same period demonstrated that patients with a TTP-like syndrome more frequently had unrelated and/or HLA-mismatched donors, and had also experienced more serious complications: grade III-IV acute GVHD and systemic bacterial, fungal, and viral infections. Three months after the diagnosis of the TTP-like syndrome, only four of 17 patients (24%) were alive; currently only one patient survives. These data emphasize: (1) the diagnosis of TTP following BMT is uncertain because of the presence of multiple BMT-associated complications. (2) The outcome of patients with TTP-like syndromes following BMT is poor. (3) Urgent intervention with plasma exchange when TTP is suspected following BMT may not always be appropriate. Alternative explanations for the signs and symptoms should be considered and treated aggressively.

Subzero temperature quenching and electrophoretic methods for the isolation of protein reaction intermediates.

A quenching technique for the study of rapid protein reactions is described which consists of injecting a small volume of aqueous solution of reactants into a large volume (X10) of hydro-organic solvent cooled at subzero temperature and mechanically shaken. The protein reaction intermediates, stabilized at subzero temperature and brought into a hydro-organic solution, can then be separated by subzero temperature electrophoretic methods, such as isoelectric focusing, in the same solvent. The alkaline hydrolysis of 2,4-dinitrophenylacetate was studied by the use of this quenching technique in order to compare the quenching time and the rate constants of the reaction with those obtained by normal rapid quenching methods. It was found that first-order reactions having rate constants up to about 5 s-1 can be satisfactorily studied by this technique. The technique is not suitable for the study of faster reactions because of the high value of the quenching time (40-100 ms). The hybridization reaction of carboxyhemoglobins A and C in aqueous solution at 22 degrees C was studied as an example of the application of this quenching technique and of the isoelectric focusing method at subzero temperature to the isolation of unstable intermediates in a protein reaction.

Bilayer-micelle transition in phosphatidylcholine-sulfatide mixtures.

Sulfatides are membrane-bound glycosphingolipids which tend to associate in micellar forms in water. In this study, combining the data obtained by several techniques, including 31P-NMR, DTA calorimetry, freeze-fracture electron microscopy, trapped volume and turbidity measurements plus enzymatic determination of outer-side "marker ganglioside", have enabled us to establish that bilayered liposomes of phosphatidylcholine formed in the presence of increasing amounts of sulfatide are stable up to 30 mol % glycolipid. Above thus, bilayered lipids progressively start to break up into micellar forms with bilayer-micelle transition complete at sulfatide concentrations above 80 mol %. The gel-to-liquid-crystalline phase transition of a dipalmitoylphosphatidylcholine-sulfatide dispersion is shown to strongly influence the equilibrium between micellar and bilayered forms, the micelles being present at higher concentrations as the fluidity of the system decreases. The possibility that such structural transitions may occur in vivo and effectively contribute to the modulation of some biological properties of the membranes is discussed.

Thrombotic thrombocytopenic purpura-hemolytic uremic syndrome: diagnosis and treatment.

Prompt recognition of thrombotic thrombocytopenic purpura-hemolytic uremic syndrome (TTP-HUS) and initiation of plasma exchange treatment is critical as it substantially decreases mortality. Nevertheless, death and long-term complications remain common. The recent relaxation of diagnostic criteria has dramatically increased the number of patients treated for clinically suspected TTP-HUS.

Associations between number of lifetime partners and other health behaviors.

OBJECTIVE: To examine associations between health behaviors and lifetime sexual partners among college students. METHODS: Data from the 1995 National College Health Risk Behavior Survey were analyzed. RESULTS: After adjusting for age, sex, and race, having 2 or more lifetime sexual partners was associated with infrequently using seat belts, driving after drinking, having a physical fight, considering suicide, and using chewing tobacco and marijuana. Significant sex interactions occurred with cigarette smoking and fruit and vegetable consumption, and significant age interactions occurred with binge drinking. CONCLUSIONS: Having multiple lifetime sexual partners (> or = 2) was associated with several negative health behaviors.

Stability and convergent validity of the Physical Activity Scale for the Elderly (PASE).

AIM: The purpose of this study was to assess the stability and convergent validity of the Physical Activity Scale for the Elderly (PASE) among rural, community dwelling elderly persons using Computer Science and Applications, Inc. Actigraph Monitors (Actigraph) as the direct criterion measure. EXPERIMENTAL DESIGN: a correlational design was employed. SETTING: rural community in the United States. PARTICIPANTS: 56 subjects (age=75.7+/-7.9 years) who were living independently and volunteered to participate in the study. MEASURES: subjects wore an Actigraph monitor during all waking hours for 7 consecutive days. At the conclusion of the 7 days, each subject met with a trained interviewer to complete the PASE questionnaire. Three days later the subjects met with the same interviewer to complete the PASE a 2nd time recalling their physical activity for the same 7-day period. RESULTS: Actigraph data indicated that subjects averaged 168.1+/-76.3 counts x minute(-1) during the 7-day period. A high intraclass correlation coefficient (r=0.91) was calculated between the 1st interview total PASE score (115.97+/-59.91) and the 2nd interview total PASE score (115.71+/-50.97). In addition, there was a statistically significant Spearman correlation coefficient of 0.43 (p<0.01) between Actigraph mean counts x minute(-1) and 1st interview total PASE scores. CONCLUSION: In this rural elderly sample, the PASE was a stable instrument with validity indices similar to those previously reported in younger, more active, populations.

The Oklahoma Thrombotic Thrombocytopenic Purpura-haemolytic Uraemic Syndrome Registry. A model for clinical research, education and patient care.

The Oklahoma Thrombotic Thrombocytopenic Purpura-Haemolytic Uraemic Syndrome (TTP-HUS) Registry has a 24 year record of success for collaborative clinical research, education, and patient care. This article tells the story of how the Registry began and it describes the Registry's structure and function. The Registry provides a model for using a cohort of consecutive patients to investigate a rare disorder. Collaboration between Oklahoma, United States and Bern, Switzerland has been the basis for successful interpretation of Registry data. Registry data have provided new insights into the evaluation and management of TTP. Because recovery from acute episodes of TTP has been assumed to be complete, the increased prevalence of hypertension, diabetes, depression, and death documented by long-term follow-up was unexpected. Registry data have provided opportunities for projects for students and trainees, education of physicians and nurses, and also for patients themselves. During our follow-up, patients have also educated Registry investigators about problems that persist after recovery from an acute episode of TTP. Most important, Registry data have resulted in important improvements for patient care.

A matched case-control study: investigating the relationship between youth assets and sexual intercourse among 13- to 14-year-olds.

BACKGROUND: The number of teenagers engaging in sexual intercourse has consistently changed over the past several years. This can become a major challenge when trying to determine why teens continue to have sex. The purpose of this paper was to investigate the relationship between youth assets and sexual intercourse among 13- to 14-year-olds. METHODS: Cross-sectional data were collected using in-person at-home interviews from a random sample of inner-city neighbourhood 13- to 14-year-old teenagers (n = 64 matched pairs; 36% 13-year-olds, 53% female, 45% white, 28% African-American, 16% Hispanic and 11% Native American). Nine individual youth assets were analysed using conditional logistic regression (matching for several demographic variables) to assess the association between assets and abstinence in younger teens. RESULTS: Conditional logistic regression analyses showed that two assets, positive Peer Role Models [odds ratios (OR) = 4.67, 95% confidence intervals (CI) = 1.93, 11.27] and Use of Time (Religion) (OR = 2.20, 95% CI = 1.04, 4.65), were significantly related (P < 0.05) to younger teens never having engaged in sexual activity. CONCLUSION: These findings suggest, when considering abstinence behaviour, involvement in religious activities and having positive peer role models appear to be protective factors related to delaying sexual intercourse among teens aged 13-14 years.

Associations between youth assets and sexual activity: does adult supervision play a role?

BACKGROUND: Youth participation in sexual risk behaviours continues to be a critically important public health topic. Additionally, as youth are frequently being left alone during the day without adult supervision, there are increased opportunities for sexual risk-taking behaviour. This study examined how the relationships of nine youth assets and sexual activity may vary according to the stratification of youth into two groups: self-care and supervised. METHODS: Data were collected through at-home, in-person interviews from a random sample of inner-city youth (mean age = 15.4 years; 51.5% female; 48.8% White; 22.4% Black; 18.5% Hispanic; 10% Native American) and their parents (n = 1079 parent/youth pairs). Nine youth assets were analysed using multiple logistic regression. Examples of assets youth may possess are: positive role models, family communication, school connectedness, constructive use of time and aspirations for the future. The item used to assess sexual intercourse was 'Have you ever had sexual intercourse ("done it", "had sex", "made love", "gone all the way")?'. Asset/risk behaviour associations that were unique to one of the two strata were the focus of the study. RESULTS: Thirty-seven per cent of youth spent two or more hours per day home alone. Youth who were supervised had a greater number of unique significant associations between assets and sexual activity than youth who were in the self-care group. CONCLUSIONS: Youth in supervised settings may be less likely to participate in sexual activity because of the presence of assets. Certain assets may also be important in deterring sexual activity for youth who are in self-care.

Participation in the transcendental meditation program and frontal EEG coherence during concept learning.

This study assesses variation in frontal bilateral EEG coherence among normal subjects during trials of a concept learning task; the task used a concept-reversal paradigm found from prior research to distinguish frontal lobe patients from normal adults. Subjects were either participants in the Transcendental Meditation (TM) program or controls matched for age, sex, and intellectual ability, and additional experimental factors were whether or not the subject gained information on a given trial and whether or not the trial occurred before, during, or after the shift of concept. It was hypothesized that: (1) higher frontal EEG coherence (alpha and beta frequencies) would be associated with trials on which information was gained; (2) higher coherence in the same frequencies would be found in the two concept-solution periods in contrast to the concept-reversal period that divided them; and (3) these patterns would be more clearly expressed among TM program participants. Each hypothesis received partial support. The first hypothesis was true only for TM program participants for alpha coherence, and only during the first concept-solution period for beta coherence. The second hypothesis was true for alpha coherence only, and the third hypothesis received support for alpha coherence. Results were not attributable to muscle or eye artifacts. However, a different response style was found to the change in concept among the two groups; control subjects displayed greater arousal (muscle artifact) during the concept-reversal period, while TM program participants displayed less arousal.