PCR-based methylation testing for Prader-Willi or Angelman syndromes using archived fixed-cell suspensions

All Prader-Willi syndrome (PWS) and 75% of Angelman syndrome (AS) patients have specific DNA methylation pattern alterations that can be used for diagnostic evaluation. The methylation testing identifies a significantly higher proportion of patients as compared to fluorescence in situ hybridization (FISH)-based microdeletion analysis and is thus a useful diagnostic evaluation for clinically suspect, but FISH-negative, patients. We used two independent PCR-based protocols for methylation testing on fixed cell specimens archived after FISH analyses. Changes in DNA methylation due to the procedure of cell fixation were ruled out by testing control specimens before and after fixation. Then methylation testing was carried out on 20 standard fixed-cell supsensions from people suspected for PWS or AS. These fixed specimens were stored after negative FISH analysis for up to 4 years at 4 degrees C in 3:1 methanol/acetic acid. Methylation patterns associated with AS (one specimen) and with PWS (one specimen) were identified for both protocols. The observed methylation patterns were concordant with the phenotypes of the positive individuals and for the two protocols used. We have, thus, shown that archived fixed-cell suspensions from individuals suspected as PWS or AS that were negative for cytogenetic/FISH microdeletions, can now be re-evaluated with PCR-based methylation testing without the need for additional blood samples from the previously studied individuals.     

The Role of Recombinant Human Thyrotropin for Diagnostic Monitoring of Patients with Differentiated Thyroid Cancer

ObjectiveTo describe the evolving role of recombinant human thyrotropin in the diagnostic evaluation of patients treated for differentiated thyroid carcinoma.MethodsA systematic review was performed of published English language articles appearing in PubMed using terms “recombinant thyrotropin” and “thyroid cancer”. The author selected articles for inclusion based upon potential for clinical impact of the reported findings.ResultsThe addition of recombinant human thyrotropin to diagnostic testing replaced the requirement for thyroid hormone withdrawal and symptomatic hypothyroidism that had been necessary to generate sufficient endogenous thyrotropin for radioiodine scanning and thyroglobulin testing. The high negative predictive value of stimulated thyroglobulin testing removed the need for serial radioiodine scanning for many patients, but repeated stimulated testing rarely appeared to add significantly. The development of highly sensitive second generation thyroglobulin assays may replace the need for stimulated testing in a subset of patients.ConclusionRecombinant human thyrotropin-stimulated testing continues to be a valuable component of follow-up testing in the first year after initial treatment of differentiated thyroid cancer. (Endocr Pract. 2013;19: 157-161)     

Technical aspects of the use of cytopathological specimens for diagnosis and predictive testing in malignant epithelial neoplasms of the lung

Lung cancer is a leading cause of cancer mortality worldwide but recent years have seen a rapidly rising proportion of cases of advanced non-small cell carcinoma amenable to increasingly targeted therapy, initially based on the differential response to systemic treatment of tumours of squamous or glandular differentiation. In two-thirds of the cases, where patients present with advanced disease, both primary pathological diagnosis and biomarker testing is based on small biopsies and cytopathological specimens. The framework of this article is an overview of the technical aspect of each stage of the specimen pathway with emphasis on maximising potential for success when using small cytology samples. It brings together the current literature addressing pre-analytical and analytical aspects of specimen acquisition, performing rapid onsite evaluation, and undertaking diagnostic and predictive testing using immunocytochemistry and molecular platforms. The advantages and drawbacks of performing analysis on cell block and non-cell block specimen preparations is discussed.     

Erectile dysfunction and diabetes mellitus

Background: Erectile dysfunction (ED) is highly prevalent, affecting ≥50% of men with diabetes mellitus (DM) worldwide.Objective: This article reviews current knowledge on the epidemiology and underlying pathophysiology of ED in men with DM, diagnostic modalities, and treatment options.Methods: A MEDLINE literature search was conducted for articles published in English from inception of the database through November 2008, using the terms erectile dysfunction, diabetes, epidemiology, pathophysiology, phosphodiesterase inhibitors, intracavernosal injection, and penile prosthesis. Data on the epidemiology, diagnosis, and treatment of ED were extracted from all relevant articles.Results: The literature search revealed 685 original articles and reviews, 67 of which were selected for inclusion in this review. DM may cause ED through a number of pathophysiologic changes, including neuropathy, endothe-lial dysfunction, cavernosal smooth muscle structural/functional changes, hormonal changes, and psychological effects. The diagnosis of ED in men with DM is based on their sexual and medical histories and results of validated questionnaires such as the International Index of Erectile Function. Laboratory examinations are usually limited to testosterone and prolactin levels that may independently contribute to ED because specialized examinations are not necessary in most diabetic men with ED. The first step in the treatment of ED in men with DM includes glycemic control and treatment of diabetic comorbidities. The associated hypogonadism must also be treated; otherwise, pharmacologic treatment may be less efficacious or not efficacious at all. Phosphodiesterase type-5 (PDE-5) inhibitors have revolutionized the treatment of ED, and they are considered first-line treatment, with a mean efficacy rate of 50% and a favorable safety profile. Intracavernous administration of vasoactive drugs is the second-line medical treatment when PDE-5 inhibitors have failed. Alprostadil is the most widely used drug for this condition, but the combination of papaverine, phentolamine, and alprostadil represents the most efficacious pharmacologic treatment option for patients whose ED does not respond to monotherapy. Excellent functional and safety results have been reported for penile prosthesis implantation, and this approach, along with proper counseling, can be considered for selected patients with treatment-refractory ED.Conclusions: ED is common in men with DM, who represent one of the most difficult-to-treat subgroups of ED patients. PDE-5 inhibitors are the first-line treatment option, followed by intracavernosal injections and implantation of a penile prosthesis.     

Interpretation of diagnostic testing strategies for production and economic usefulness

Veterinarians providing reproductive services use a variety of diagnostic testing methods, including physical examination, laboratory testing, diagnostic imaging, and performance record evaluation. The diagnostic end point may be a physical diagnosis of pregnancy, attainment of puberty, or adequate quality and quantity of sperm; furthermore, it may be a medical diagnosis of reproductive tract pathology, presence of an infectious pathogen, or abnormal hormonal status. Proper interpretation of test results requires an understanding of how sensitivity and specificity (as measures of test accuracy), and prevalence of the condition, affect the interpretation of an individual result. For many diagnostic questions, the proper use of more than one test, either in series or in parallel, allows veterinarians to optimize their diagnostic accuracy and the economic return for the testing strategy.     

Single slide assessment: A highly effective cytological rapid on‐site evaluation technique for endobronchial and endoscopic ultrasound‐guided fine needle aspiration

No standardised, comprehensive approach to rapid on‐site evaluation (ROSE) of cytology samples currently exists. Recent meta‐analysis indicates variation in the effectiveness of ROSE, however, reviews commonly omit the details of how ROSE is conducted. This review demonstrates the clinical effectiveness of single slide assessment (SSA) for ROSE of cytology samples, providing a highly effective, standardised methodology, maximising cell yield and the diagnostic potential of samples obtained via endobronchial or endoscopic ultrasound. Advances in molecular testing and immunotherapy now allow patients to access sophisticated, targeted cancer treatments and, consequently, obtaining diagnostic material alone is no longer sufficient. SSA uses specific criteria, based on the morphological presentation, to ensure sufficient material is obtained through one procedure, allowing for all the molecular profiling and tumour expression testing required to provide the patient and clinicians with the optimal treatment options. In total, 450 endobronchial or endoscopic ultrasound procedures were conducted with ROSE SSA performed by a biomedical scientist between 2010 and 2017. In 97% of cases, ROSE SSA matched the final report (inadequate vs adequate—benign material vs malignancy). ROSE SSA provided sufficient material for immunocytochemistry in 200/208 cases (96%) and for additional molecular testing/tumour profiling in 92% (85/92) of cases. The median number of needle passes was three. ROSE SSA streamlines diagnostic pathways; minimising risk of complications to patients, reducing cost and delays to treatment associated with repeat or more invasive procedures. Using SSA, sufficient material for a comprehensive diagnosis can be obtained in one procedure.     

Penile revascularization in the treatment of vasculogenic impotence

Arterial insufficiency is the most common organic cause of impotence. The diagnosis can be established on a clinical basis through the use of Doppler probe evaluation of the penile pulses, penile blood pressure measuring, and nocturnal penile tumescence monitoring. The diagnosis may be confirmed by internal pudendal arteriography. Through microsurgical arterial revascularization, a 60 percent long-term success rate has been achieved. Two early and two late failures suggest the need for improved patient selection and confirm the progressive nature of the arteriosclerotic process.     

Methodologic evaluation of the lumbar disc syndrome.

Though the lumbar disc syndrome is a costly and ubiquitous affliction, effective evaluation of the disease process has been confounded by major unaddressed methodological short falls. Prominent difficulties include: inattention to the clinical boundaries of the syndrome, neglected co-morbid disease processes, comparison of unequal treatment groups and premature clinical data extrapolation, inadequate diagnostic validation, variability in surgical observation, and reliance upon follow-up techniques faulted by unaddressed distorting factors. Proposals for improvement include: formulation of suitable stratification subgroups emphasizing age and sign-symptom intensity and duration, techniques for improved diagnostic return from surgical exploration, suggestions toward improved quantitation of clinical testing procedures, and implantation of a quality of life scale.     

Penile prosthesis implantation for end-stage erectile dysfunction after radical prostatectomy

When erectile dysfunction occurs after radical prostatectomy and phosphodiesterase-5 inhibitor therapy fails, second-line therapies such as vacuum constriction devices, intraurethral prostaglandins, and penile injection therapy should be offered. When second-line therapies are not effective or acceptable to the man and his partner, penile prosthesis implantation becomes the treatment of choice. Today's 3-piece inflatable devices offer flaccidity and erection that approach the natural state. Design improvements have resulted in devices that have freedom from mechanical failure ranging from 92% to 94%. Antibiotic and hydrophilic coatings have reduced infection rates.     

Penile cancer: is there an epidemiological role for smoking and sexual behaviour?

A retrospective study was carried out to determine whether penile cancer, like cervical cancer, was associated with smoking and sexual behaviour. Altogether 244 men with penile cancer and 232 matched controls completed a questionnaire by post or telephone. Data on marital state, socioeconomic group, occupation, history of phimosis and balanitis, sexual behaviour, and smoking were obtained. The results of statistical analyses confirmed that phimosis and balanitis were risk factors for penile cancer, but there was no epidemiological evidence for it being a sexually transmitted disease. Smoking was a risk factor with a dose-response relation and remained associated with penile cancer even after adjustment for confounding factors. Penile cancer is associated with smoking independently of phimosis; treatment of phimosis alone does not remove the risk caused by smoking.     

Treatment of Peyronie's disease by dermal grafting.

We describe our experience with dermal grafting for the treatment of Peyronie's disease in 27 patients over a 3-year period. With the various nonsurgical treatments, a significant number of patients remain handicapped by the disease. For this unresponsive group, we recommend surgical excision of the penile plaque with dermal grafting. The proper selection of patients for this operation is of paramount importance, and this can be facilitated by a preoperative psychological evaluation in doubtful cases.     

Long-term follow-up of total penile reconstruction with sensate osteocutaneous free fibula flap in 18 biological male patients

Surgical reconstruction of the penis is challenging because of the many cosmetic and functional (e.g., sexual intercourse and voiding) requirements that must be addressed. Since the free sensate osteocutaneous fibula flap was first described for total penile reconstruction in 1993 it has been widely accepted, with its advantages and minimal shortcomings. In this article, the authors present the longest follow-up of biologically male patients with free fibular phalloplasties. Since 1994, 18 biologically male patients with total penile losses for various reasons were treated with free sensate osteocutaneous fibula flaps. All patients were included in the study. The ages of the patients ranged between 20 and 26 years (mean, 22.2 years). The average follow-up period was 5.4 years (range, 1 to 9 years). Patient satisfaction was evaluated by a questionnaire regarding both quality of orgasm and daily activities. Conventional radiographic imaging, magnetic resonance imaging, and bone mineral densitometry were performed to evaluate the fate of the bony component of the flap. Also, sensibility was evaluated by bulbocavernous reflex and penile somatosensory evoked potentials testing in nine patients. Six patients married, and five of them had six children. Most patients and their partners reported pleasurable sexual intercourse and orgasm. Conventional radiographs of the fibular bone in neophalluses showed robust, calcified bone structure without any evidence of bone resorption or fracture. The magnetic resonance images showed the cortical substance and spongiosum of the bone marrow, which are characteristic signs of bone viability. After intravenous injection of gadolinium, the neophallus bone showed uptake of contrast medium. Viability of neophallus bone was shown even at 9-year follow-up (the longest follow-up in the literature). Dual energy x-ray absorptiometry measurements of the penile bone grafts showed that fibular components in the penis had bone mineral density values that were close to but lower than those of intact fibula in the same subjects. These results were considered as evidence of viability of bone grafts. Neural integrity was found between the nerves of the neophallus and the residual penile bodies by both bulbocavernous reflex and penile somatosensory evoked potentials tests. In conclusion, free sensate fibula flap phalloplasty provides the cosmetic and functional requirements that an ideal penis should have. All results put an end to the discussion that the fibular component of the neophallus could resorb. Constitution of neural integrity is important in terms of pleasurable sexual intercourse. The authors believe the free sensate osteocutaneous fibula flap should be considered as the standard in penile reconstruction.     

Applying random forests to identify biomarker panels in serum 2D-DIGE data for the detection and staging of prostate cancer

In recent years, Prostate Specific Antigen (PSA) testing is widespread and has been associated with deceased mortality rates; however, this testing has raised concerns of overdiagnosis and overtreatment. It is clear that additional biomarkers are required. To identify these biomarkers, we have undertaken proteomics and metabolomics expression profiles of serum samples from BPH, Gleason score 5 and 7 using two-dimensional difference in gel electrophoresis (2D-DIGE) and nuclear magnetic resonance spectroscopy (NMR). Panels of serum protein biomarkers were identified by applying Random Forests to the 2D-DIGE data. The evaluation of selected biomarker panels has shown that they can provide higher prediction accuracy than the current diagnostic standard. With careful validation of these serum biomarker panels, these panels may potentially help to reduce unnecessary invasive diagnostic procedures and more accurately direct the urologist to curative surgery.     

Les traumatismes de la verge: à propos de 23 cas

The authors report 23 cases of penile injuries based on a retrospective study of their urological practice in Senegal. The various cases were distributed as follows: fracture of the penis (19 cases), corpora cavernosa and urethral gunshot injuries (2 cases), rupture of the superficial dorsal vein of the penis (1 case) and laceration of the penile skin (1 case). The mean age of these patients was 32.4 years. Early surgical treatment of all penile fractures reduces the complication rate.     

Penile paralysis and paraphimosis associated with reserpine administration in a stallion

Reserpine was administered to an 8-yr-old Thoroughbred stallion at a dosage of 5 mg subcutaneously (s.c.) every 2 wk for a 2-mo period to control unmanageable behavior. Reserpine produced a satisfactory calming effect that lasted for about 2 wk. After the last injection, the stallion developed penile paralysis and was unable to retract his penis, resulting in paraphimosis and attendant penile edema. The prolapsed penis was reduced and kept within the prepuce by placing a purse string suture in the preputial orifice. Phenylbutazone was given orally (1 gm) and the stallion was exercised for 20 to 30 min twice daily. This treatment was continued for 20 d with little improvement. The purse string retention suture cut through the skin 5 d after the stallion was discharged from the clinic. The penis was then supported against the abdominal wall and the stallion was exercised by hand for 30 min each day. The stallion was not used for breeding within 34 mo after the last injection of reserpine. A breeding soundness examination was performed approximately 3 yr after the initial injury. At this time the stallion's penis was noted to extend 5 to 8 cm from the prepuce when in a detumescent state. Although the stallion protruded his penis when exposed to a mare in estrus, a full rigid erection was never attained. Examination of the penis revealed partial engorgement of the corpus cavernosum penis and a 2.5-cm-wide dorsal semi-circumferential depression of the penile shaft approximately 10 to 12 cm proximal to the glans penis. The penile shaft and glans penis distal to this depression were cooler than the proximal portion of the penis. Semen collection was attempted, aided by manual insertion of the penis into the artificial vagina. When serving the artificial vagina, no "belling" of the glans penis was observed, although ejaculation occurred. Semen evaluation indicated normal spermatozoal motility and morphology parameters. The stallion was able to breed several mares with manual assistance to guide the penis into the vagina and one mare was diagnosed pregnant.     

Amputation du gland au cours de la circoncision et réimplantation : à propos d’un cas et revue de la littérature

Penile amputation due to circumcision is a tragic complication in which the operator is responsible. The current treatment is based on microsurgical replantation methods by anastomosing penile dorsal vessels and nerves. We report a new case of penile glans amputation due to circumcision in a 6-year-old boy. Replantation was done without microvascular and nerves anastomosis. After 7 months of treatment, the final result was found to be good in terms of the urinary stream, erectile function, sensitivity and morphological aspect of the glans.     

Peyronie's Disease: A Review

Peyronie's disease is an acquired benign condition without known systemic sequelae with presenting symptoms that include the presence of a plaque or induration of the penile shaft, penile curvature or deformity during erection, penile pain, and erectile dysfunction. This article reviews the natural history of the disease, discusses the disease's etiology (widely thought to involve minor penile trauma with subsequent aberrant wound healing), and outlines proper clinical evaluation of Peyronie's disease patients. Medical treatments can be systemic (colchicine, potassium aminobenzoate, vitamin E), intralesional (steroids, verapamil, collagenase, interferons), or topical. Surgical therapy for Peyronie's disease (plication, graft-based, and prosthetic techniques) should be reserved for the man who has failed conservative therapy and whose curvature, indentation, or erectile dysfunction precludes intercourse. Regardless of the surgical procedure, the patient should be made aware of the inherent risks of surgery.     

La place des protheses de verge dans le traitement de la maladie de La Peyronie

In Peyronie’s disease, a penile curvature makes sexual intercouse painful or impossible. Surgery is a treatment option after medical treatment failure. The aim of surgery is to restore an erection allowing sexual intercourse. Beside corpus cavernosus modeling procedures, penile prosthesis implantation (i.e.,semi-rigid, inflatable, orimplants) is rarely indicated. Such procedures should be based on erectile response to intracavernous injection and penile and corpus cavernosus anatomy.     

Collection and quality of rhesus monkey semen

Electroejaculation is an accepted method of semen collection from nonhuman primates. Although both penile and rectal probe stimulation techniques have been used, there has been a general lack of consistency and detail regarding their application. This report describes the collection, processing, and evaluation of rhesus monkey semen contrasting two methods of penile electroejaculation: 1) a constant-voltage method where stimulus current is a variable and 2) a constant-current method where stimulus current is operator-controlled. The constant-current method was the more efficient procedure, requiring a lower stimulus current for successful electroejaculation. The influence on semen quality of potentially toxic agents used in the procedure, surgical glove powder and electrolyte cream, was tested; both were detrimental as measured by motility loss. No correlation was found between coagula volume and sperm numbers. The intra- and interanimal variability in semen samples from six monkeys was also evaluated. Penile electroejaculation, combined with control of stimulus current, provides a consistent, successful, and humane method for the collection of semen in the rhesus monkey.     

Clinical Outcomes and Correlates of TP53 Mutations and Cancer

The initial observation that p53 accumulation might serve as a surrogate biomarker for TP53 mutation has been the cornerstone for vast translational efforts aimed at validating its clinical use for the diagnosis, prognosis, and treatment of cancer. Early on, it was realized that accurate evaluation of p53 status and function could not be achieved through protein-expression analysis only. As our understanding of the p53 pathway has evolved and more sophisticated methods for assessment of p53 functional integrity have become available, the clinical and molecular epidemiological implications of p53 abnormalities in cancers are being revealed. They include diagnostic testing for germline p53 mutations, and the assessment of selected p53 mutations as biomarkers of carcinogen exposure and cancer risk and prognosis. Here, we describe the strengths and limitations of the most frequently used techniques for determination of p53 status in tumors, as well as the most remarkable latest findings relating to its clinical and epidemiological value.