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Biomedical subjects

H John

Publications and source records attributed to H John.

At least 19 recordsLinked to original sources
PubMed2006

Expression of smoothelin in the normal and the overactive human bladder.

PURPOSE: We established the expression pattern of smoothelin, a marker protein for contractile smooth muscle cells, in the human detrusor and investigated its possible impact on bladder overactivity. MATERIALS AND METHODS: Detrusor samples of 13 overactive bladders (sensory urge and detrusor instability) were obtained before botulinum toxin injection and compared to those of 8 normally contractile, nonobstructed bladders obtained during radical cystectomy. Smoothelin mRNA expression patterns were investigated by Northern blot and variant specific reverse transcriptase-polymerase chain reaction as well as by quantitative reverse transcriptase-polymerase chain reaction on laser capture, microdissected smooth muscle. At the protein level smoothelin was investigated by standard and quantitative immunohistochemistry. RESULTS: The bladder muscularis expressed vascular and visceral smoothelin isoforms, and 2 of the known splice variants. In the smooth muscle of patients with detrusor instability and sensory urge a significant 2.4 and 2.2-fold increase, respectively, in smoothelin variant 1 mRNA was observed in comparison to that of normal controls. Analyses at the smoothelin protein level confirmed significant up-regulation in these bladder dysfunctions by a factor of 2.3 and 1.8, respectively. No significant difference in smoothelin expression was observed between detrusor instability and sensory urge. CONCLUSIONS: Increased expression of smoothelin in patients with detrusor instability and sensory urge implies that the etiology of these dysfunctions includes changes in myogenic parameters. In addition, our data support the new classification of the International Continence Society for overactive bladder proposing that sensory urge and detrusor instability represent a single clinical entity.

Cytoskeletal Proteins↗
PubMed2006

Experience with 100 cases treated with botulinum-A toxin injections in the detrusor muscle for idiopathic overactive bladder syndrome refractory to anticholinergics.

PURPOSE: In this prospective, nonrandomized, ongoing study we evaluated the efficacy and safety of botulinum-A toxin injections in the detrusor muscle to treat patients with idiopathic overactive bladder resistant to conventional treatment, such as anticholinergic drugs. MATERIALS AND METHODS: A total of 23 men and 77 women with a mean age of 63 years (range 24 to 89) with nonneurogenic overactive bladder, including urgency-frequency syndrome, and incontinence despite the administration of maximal doses of anticholinergics were consecutively treated with injections of 100 U botulinum-A toxin in the detrusor muscle at 30 sites under cystoscopic guidance. Micturition diary, full urodynamics, neurological status and urine probes were performed in all participants before treatment. Bladder biopsies were done only in cases of suspected bladder fibrosis or unclear findings. Special attention was given to reflex volume, maximal bladder capacity, detrusor compliance, post-void residual urine, urgency and frequency/nocturia. Clinical, urodynamic and quality of life assessments were performed at baseline, and 4, 12 and 36 weeks after botulinum-A toxin treatment. RESULTS: Overall after 4 and 12 weeks 88% of our patients showed significant improvement in bladder function in regard to subjective symptoms, quality of life and urodynamic parameters (p <0.001). Urgency disappeared in 82% of the patients and incontinence resolved in 86% within 1 to 2 weeks after botulinum-A toxin injections. Mean frequency decreased from 14 to 7 micturitions daily (-50%) and nocturia decreased from 4 to 1.5 micturitions. Mean maximal bladder capacity increased 56% from 246 to 381 ml, mean detrusor compliance increased from 24 to 41 ml/cm H(2)O and pretreatment detrusor instability (mean reflex volume 169 ml) resolved in 74% of patients. Mean volume at first desire to void increased from 126 to 212 ml and mean urge volume increased from 214 to 309 ml. There were no severe side effects except temporary urine retention in 4 cases. Only in 8 patients was the clinical benefit poor and analysis revealed preoperative low detrusor compliance. Mean efficacy duration +/- SD was at least approximately 6 +/- 2 months and then symptoms began to increase. CONCLUSIONS: Our results show that intradetrusor botulinum-A toxin injections may be an efficient and safe treatment option in patients with severe overactive bladder resistant to all conventional treatments.

Adult↗
PubMed2005

[Diagnostic assessment of painless microhematuria: prospective study comparing image quality, assessibility and diagnostic certainty of multidetector-row CT and intravenous pyelography within a single examination].

PURPOSE: The purpose of this study is to prospectively compare intravenous pyelography (IVP) and combined unenhanced and excretory phase multidetector-row CT (MDCT) with respect to image quality, diagnostic certainty and diagnostic concordance with the final clinical diagnosis in patients with painless microhematuria. MATERIALS AND METHODS: Unenhanced MDCT, IVP and excretory phase MDCT were performed in 59 consecutive patients (21 women, 38 men, mean age 56 +/- 19 years, range 23 - 83 years) with painless microhematuria of unknown origin during a single examination with a single contrast media application (100 ml, non-ionic iodinated contrast media). Images were assessed by two experienced urogenital radiologists in consensus for image quality, diagnostic certainty of stone detection, obstruction, parenchymal lesions and morphological distinctive features. Imaging diagnoses of MDCT and IVP were compared with the final clinical diagnoses. In case of failure to detect an relevant pathology, the final clinical diagnosis was established after a mean follow-up period of 18 +/- 6 months (10 months to 2 years). Costs and radiation exposure of IVP and MDCT were compared. RESULTS: MDCT scan performed better than IVP in terms of image quality for all regarded variables. Image quality of MDCT was rated in all parameters as very good or good; the image quality of IVP differed in a wide range. MDCT and IVP reached a sensitivity of 100 % and 50 % for stone detection (n = 14, p = 0.008), respectively. Two bladder stones were not detected by IVU but correctly seen with MDCT. MDCT and IVP were unsatisfactory for detecting transitional cell carcinomas (n = 4, 2 of 4 detected with MDCT, 0 of 4 detected with IVU). One false positive transitional cell carcinoma was detected with IVP, none with MDCT. Additional relevant pathological changes (one teratoma, one abdominal aortic aneurysma and one abscess) were detected using MDCT but missed with IVP. In 38 of 59 patients (64 %) imaging and clinical follow-up over up to 24 months did not reveal any pathology to explain the microhematuria. The costs of the IVP (283 Euro) were lower compared with non-enhanced MDCT (380 Euro) or combined non-enhanced and contrast-enhanced MDCT (560 Euro). The radiation exposure was 23 - 27 mSv for MDCT and 2.3 mSv for IVP. CONCLUSION: MDCT performed better regarding image quality, subjective diagnostic certainty and diagnostic results with respect to stone detection. Since urolithiasis is a frequent cause of painless microhematuria MDCT is recommended as the initial imaging modality rather than IVU.

Adult↗
PubMed2003

Characterization of human circulating TIG2 as a ligand for the orphan receptor ChemR23.

The orphan receptor ChemR23 is a G-protein coupled receptor (GPCR) with homology to neuropeptide and chemoattractant receptors. Tazarotene, a synthetic retinoid activating retinoic acid receptor (RAR), up-regulates tazarotene-induced gene-2 (TIG2). The function and molecular target of this protein are now described. By means of reverse pharmacology screening using a peptide library generated from human hemofiltrate, we have isolated and identified TIG2 as the natural ligand of ChemR23 and report the specific molecular form of the bioactive, circulating TIG2, representing the amino-acid residues 21 to 154 of the 163 amino acid-containing prepropeptide. Based on the expression pattern of ChemR23 and TIG2, the physiological role in bone development, immune and inflammatory responses and the maintenance of skin is now being investigated.

Amino Acid Sequence↗
PubMed2003

[Diagnosis of chronic prostatitis: 4 or 2 glass sample?].

Chronic pelvic pain syndrome is still an important clinical problem. The NIH-prostatitis classification introduced 1998 for diagnosis and treatment measures is based on extended microbiological analysis of urine, expressed prostate secretion (4-glass test), and ejaculate. We investigated if a simple urine analysis of an urine sample before and after prostatic massage (2-glass test) could replace the 4-glass test. 143 patients with the diagnosis chronic prostatitis were included in this prospective study. The results showed that the expensive and time consuming 4-glass test can be replaced by a simple 2-glass test. Extended examinations should only be performed in special cases.

Adult↗
PubMed2003

[Four-glass or two glass test for chronic prostatitis].

Chronic pelvic pain syndrome is still an important clinical problem. The NIH prostatitis classification introduced in 1998 for diagnosis and treatment measures is based on extended microbiological analysis of urine and expressed prostate secretion (4-glass test). In 1997 J.C. Nickel proved that the culture and microscopic examination of urine before and after prostatic massage leads to the same results as the 4-glass test. In our prospective study on 143 patients with a diagnosis of chronic prostatitis, we analyzed this statement and came to the same results. We therefore recommend replacing the expensive and time-consuming 4-glass test by a simple preprostatic and postprostatic massage urine culture. Further examinations should only be performed in special cases.

Adult↗
PubMed2003

[Urinary incontinence in the man--pathophysiology and diagnosis].

About 7-8% of men experience unvoluntary urinary leak. Incidence in patients older than 60 years is 2-3 fold increased. Urge incontinence is the mostly present, less frequent are overflow-incontinence/chronic retention and urinary stress incontinence. In men, prostate and bladder dysfunction as well as neurologic diseases are responsible for urinary incontinence. The baseline diagnostics include micturition protocols, the urologic clinical examination, residual urine determination and laboratory analyses of urine and blood. Extended diagnostics proceed to morphological and infectious etiologies and base on urodynamic evaluation.

Aged↗
PubMed2003

[Conservative and surgical therapy of urinary incontinence and bladder complaints in the man].

Treatment of incontinence and bladder complaints in the male should be directed to the cause whenever possible. Frequently, however, only symptomatic therapy is possible. Urge incontinence or overactive bladder due to obstruction should primarily be treated by eliminating the obstruction. Medical and surgical treatment methods are available for benign prostatic hyperplasia, bladder neck hypertrophy and prostatic cancer. In contrast, bladder neck sclerosis and uretheral strictures can only be treated surgically. Anticholinergics are primarily indicated if urge symptoms/incontinence persist after obstruction has been relieved or if urge incontinence occurs without obstruction. Seldom, in special cases injection of Botulinustoxin A or augmentation of the bladder may be indicated. Another possible cause of urge symptoms is urinary tract infection. This should be adequately treated according to resistance studies and the cause of the infection determined. In cases of overflow incontinence the infravesicle obstruction must be sought and treated. If limited detrusor contractability is the cause of overflow incontinence and the bladder cannot be emptied through pressmicturition, parasympathicometics may be of help. By insufficient effect, the procedure of intermittent self-catheterization must be taught. If this is not possible, the last resort is placement of a transuretheral or percutaneous catheter for continuous drainage. Stress incontinence is a rare complication in men, usually following prostatic surgery. It can be treated conservatively with pelvic floor training and alpha-adrenergic receptor agonists and if necessary surgically with submucosal collagen or silicon injections in the sphincter area or implantation of a sphincter prosthesis. Supravesicular urinary diversion is occasionally necessary after conservative and less invasive surgical measures have been exhausted and symptomatic suffering persists. Neurogenic disturbances in bladder capacity and/or emptying can be treated conservatively, medically, surgically or a combination of these depending upon the site of the lesion and the resulting urodynamic patterns.

Animals↗
PubMed2003

[Gap junctions in the human urinary bladder].

PURPOSE: Gap junctions are intercellular contacts important for the synchronization of muscle cell activity through electrical coupling. Since the role of gap junctions for the function of smooth bladder muscle is still a matter of debate, we investigated the occurrence of gap junctions and the gap junctional protein connexin (Cx) 45 in the detrusor of the nonobstructed stable human bladder. MATERIALS AND METHODS: Detrusor biopsies from 6 patients aged 64 (55-72) years with stable nonobstructed bladders were investigated for the occurrence of gap junctions by electron microscopy, molecular biological techniques and immunohistochemistry. RESULTS: Transmission electron microscopy and freeze fracture showed the presence of gap junction at plasma membranes of detrusor smooth muscle cells. By reverse transcriptase (RT) polymerase chain reaction (PCR) and in situ hybridization, we found an expression of Cx45 in the detrusor. These data were confirmed by immunolocalization of Cx45 on smooth muscle cells. CONCLUSIONS: This study provides morphological as well as molecular biological and immunohistochemical evidence that bladder smooth muscle cells are electrically coupled.

Aged↗
PubMed2003

The effect of seminal vesicle-sparing radical prostatectomy on serum prostate-specific antigen level.

OBJECTIVE: To investigate if the remaining seminal vesicle tips can affect serum levels of prostate-specific antigen (PSA) in patients after seminal vesicle-sparing radical prostatectomy (SVRP). PATIENTS AND METHODS: Thirty-six patients were treated by either radical retropubic prostatovesiculectomy (23) or SVRP (13). Serum PSA was monitored in all patients before surgery, and at 6 weeks and 30 months afterward. Samples of normal seminal vesicles from radical cystectomies (six) were also snap-frozen and either processed for semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) using primers against PSA and alpha-actin (for normalization) or for PSA immunohistochemistry. RESULTS: RT-PCR and sequencing showed that the seminal vesicles synthesise PSA mRNA. Furthermore, PSA peptide was detectable in the glandular epithelium of the seminal vesicle using immunohistochemical methods. There was no significant difference in serum PSA levels after standard or SVRP, with median (range) values (ng/mL) at 6 weeks of 0.04 (0.04-0.9) and 0.04 (0.04-0.66) and at 30 months of 0.17 (0.04-3.8) and 0.22 (0.04-58.2), respectively. CONCLUSION: Although the seminal vesicles produce PSA, the PSA derived from the remaining seminal vesicle tips after SVRP has no effect on the oncological follow-up of these patients.

Age Factors↗
PubMed2003

Immunological alterations in the ejaculate of chronic prostatitis patients: clues for autoimmunity.

The aim of this prospective study was to observe immunophenotypic patterns in the ejaculate of patients with noninflammatory chronic pelvic pain syndrome (Cat IIIB CPPS) and to test for a possible autoimmune aetiology. Thirty-five patients of a total of 88 patients with chronic prostatitis Cat IIIB were consecutively selected. Monthly ejaculate testing was carried out for IgG, IgA, IgM, IL-1alpha, sIL-2R and IL-6. The control group for ejaculate analysis was composed of 96 normal ejaculates (according to the WHO criteria). Immunohistochemical detection of CD3 cells (T lymphocytes) and CD20 cells (B lymphocytes) was performed in 71 biopsy cylinders of Cat IIIB CPPS patients and in 25 prostate biopsy cylinders of subjects without symptoms or obstruction. Intra-acinar T-lymphocytic infiltrates were dominated by T-cytotoxic cells (P = 0.05). Ejaculate IL-6 and ejaculate IgA increased significantly and dropped again, correlating with a release of clinical symptoms. Inflammatory ejaculate interleukin concentrations correlated with the immunohistochemical findings with presence of large numbers of T cells (all P-values < or = 0.01). Immunomodulation was performed in a pilot series of three patients by five monthly cycles of IgG (Sandoglobulin), 1 g kg-1 body weight. Immunomodulation with IgG decreased pain moderately and did not change ejaculate interleukin and immunoglobulin concentrations. In summary, interleukin and immunoglobulin determinations in the ejaculate revealed an inflammatory process even in Cat IIIB CPPS. The findings of intra-acinar T-cell rich infiltrates and the associated inflammatory reaction may indicate a possible autoimmune component in the aetiology of CPPS. Exact origin and role of interleukin changes in the ejaculate of CPPS patients need to be further evaluated. Unfortunately, pilot series with immunomodulation with IgG do not seem to provide clear clinical benefit.

Adult↗
PubMed2003

[Diagnostic and therapeutic procedures for hemospermia].

In general, hemospermia represents a trivial condition: nevertheless, it is often alarming for the patient and his partner. Although differential diagnosis is comprehensive, the most frequent underlying causes are infection and inflammatory processes in the lower seminal passages. Cases of primary and solitary hemospermia can be adequately assessed by urinalysis, blood pressure measurement, and reassurance of the patient. Persistent and recurrent cases of hemospermia are best clarified by transrectal ultrasound examination, cystoscopy, and magnetic resonance imaging. Treatment is based on the examination results.

Cystoscopy↗
PubMed2002

[Partial androgen deficiency in the aging male: diagnosis and therapy].

After the fourth decade, androgen serum level fall while the binding capacity for testosterone, the sex hormone binding globulin, raises. Clinical symptoms and hypogonadism define the syndrome of the partial androgen deficiency of the aging male (PADAM). Testosterone substitution may be performed by intramuscular injections, oral or transdermal testosterone application. Each patient should undergo urological evaluation before starting a testosterone replacement therapy. Life style drugs should not be used according the currently rational data. Well balanced nutrition, physical training and an active social and sexual life are important factors to achieve a healthy aging.

Administration, Cutaneous↗
PubMed2002

The 15-domain serine proteinase inhibitor LEKTI: biochemical properties, genomic organization, and pathophysiological role.

Proteinases are involved in specific and non-specific proteolytic reactions, and participate in many pathophysiological processes. Normally, they are regulated by endogenously produced proteinase inhibitors which, thus, represent lead structures for the development of therapeutics. We succeeded in partially isolating and cloning a novel human serine proteinase inhibitor which, according to its structure and the expression pattern of the corresponding gene, was termed lympho-epithelial Kazal-type-related inhibitor (LEKTI). This inhibitor is of special interest because it exhibits an extraordinarily large number of 15 potentially inhibitory domains and is of pathophysiological importance for the severe congenital disease Netherton syndrome. Here, we review the as yet known data on protein structure, biochemical properties, genomic organization and gene expression. Furthermore, the relevance of LEKTI for several disorders pointing out its possible future therapeutic value, is discussed.

Animals↗
PubMed2001

Determination of the disulfide bond pattern of the endogenous and recombinant angiogenesis inhibitor endostatin by mass spectrometry.

Endostatin, a C-terminal fragment of collagen XVIII, is a promising protein drug which is in development for cancer therapy due to its anti-angiogenic activity. Although several endogenous molecular forms of human endostatin differing in their N-terminal length and their post-translational modifications (18.5-22 kDa) have been discovered, only one recombinant form of 20 kDa is used in clinical trials. This protein, recombinantly expressed in Pichia pastoris, contains four cysteines forming two disulfide bonds (Cys1-Cys4 and Cys2-Cys3). In contrast, there are conflicting data about the disulfide pattern of endogenous material. This report presents the disulfide analyses of both the endogenous circulating endostatins isolated from human hemofiltrate and the recombinant protein. The determination of the disulfide pattern was performed by Edman degradation, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and electrospray ionization ion trap mass spectrometry (ESI-ITMS) performed in the off-line nanospray mode. All native and recombinant endostatins exhibited a Cys1-Cys4 (Cys(162)-Cys(302)) and Cys2-Cys3 (Cys(264)-Cys(294)) linkage. For a clear discussion of fragmented disulfide-bridged peptide chains obtained from MS(n) experiments, a modified general nomenclature is proposed.

Amino Acid Sequence↗
PubMed2001

Identification of nucleated cells in urine using lectin staining.

Microscopic examination of urinary sediment is an integral component in the evaluation of nephropathies. However, identification and differentiation of the nucleated nonsquamous cells in urine is often difficult using such conventional techniques as phase contrast or bright field microscopy, even after Papanicolaou staining, and requires a lot of experience. We now report a method to differentiate urinary cell types using lectin staining. Twenty-five lectins were examined with respect to their binding pattern on cryosections of the human kidney and urinary tract, as well as binding to blood cells. The specificity of lectin binding to a cell type both in situ and in urine was confirmed by double labeling with specific antibodies directed against various sections of the nephron or nucleated blood cells. For urine cytologic examinations, acetone-fixed cytopreparations of urinary sediments were incubated with a combination of a fluorescein isothiocyanate (FITC)-coupled and a rhodamine-coupled lectin, followed by staining of the nuclei with 4',6-diamidino-2-phenylindole. Specimens were examined in triple immunofluorescence (FITC/rhodamine/UV). Cell types could be identified by their characteristic lectin-binding pattern. For example, the lectin combination of Sophora japonica agglutinin (aggl; SJA) and Erythrina cristagalli aggl (ECA) permitted a differentiation between cells of the proximal tubules (SJA positive [SJA+], ECA+), distal tubules (SJA negative [SJA-], ECA+), collecting ducts (SJA+, ECA-), and lymphocytes (SJA-, ECA-). In preliminary studies, examination of urinary sediment in various chronic nephropathies by this technique showed differences in their cellular excretion pattern. In summary, staining urinary sediments with combinations of lectins provides a rapid and relatively inexpensive method for a facilitated and reliable differentiation of the various nucleated cell types in urine.

Binding Sites↗
PubMed2001

Evidence of trigonal denervation and reinnervation after radical retropubic prostatectomy.

PURPOSE: Urinary incontinence continues to be a major consequence of radical prostatectomy. To understand the pathophysiology of this dysfunction we studied the impact of autonomic innervation of the superficial trigone on postoperative urinary continence. MATERIALS AND METHODS: To investigate nerve fiber density biopsies of the superficial trigone were obtained in 34 patients preoperatively as well as 6 weeks and 6 months postoperatively in 15 and 19, respectively. Specimens were Bouin fixed, paraffin embedded and processed for light microscopic immunohistochemical evaluation using an antibody against protein gene product 9.5, a general neuronal marker protein. In parallel we performed a comprehensive urodynamic evaluation, including determination of maximal urethral closure pressure and posterior urethral sensory threshold. RESULTS: Postoperatively protein gene product 9.5 immunoreactive nerve fiber density was generally decreased. However, nerve fiber density after 6 weeks of incontinence in 12 of 15 patients was only 7%, while 3 of 15 who were continent preserved 36% of initial nerve fiber density. After 6 months nerve fiber density in 19 patients increased in 3 with incontinence to 20% and in 16 with continence to 44% of intraoperative density. Urinary incontinence was associated with decreased trigonal innervation, a high sensory threshold and low maximal urethral closure pressure. CONCLUSIONS: Protein gene product 9.5 immunoreactive nerve fiber density corresponds with posterior urethral sensory threshold and urinary continence. Thus, preserving trigonal innervation and postoperative reinnervation may be important factors for achieving early postoperative urinary continence after radical prostatectomy.

Aged↗