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Biomedical subjects

H John

Publications and source records attributed to H John.

At least 37 records · Page 2Linked to original sources

Ultrastructure of the trigone and its functional implications.

OBJECTIVE: To investigate the ultrastructure of the trigonal muscle (= superficial trigone), relate it to normal function, and identify any changes it may undergo in voiding dysfunction. MATERIALS AND METHODS: 20 men (median age 67 years) with low-stage prostatic carcinoma, 10 with and 10 without bladder outlet obstruction, were selected by urodynamic evaluation. Trigonal biopsy was performed at radical prostatectomy, and processed for electron-microscopic study by standard procedures. Biopsies were evaluated independently by 2 examiners without prior knowledge of urodynamic data. RESULTS: Three obstructed and 1 unobstructed bladder had impaired detrusor contractility, and 1 obstructed bladder had detrusor overactivity. Compared to the previously investigated detrusor, the trigonal muscle had smaller compact bundles and fascicles with less collagen and more elastic tissue. Muscle cells had no or rare intermediate junctions that mediate mechanical coupling in normal detrusor, but predominant close cell appositions that mediate electrical coupling. Smooth muscle in most biopsies had widespread or focal features characteristic of the aged detrusor. None, however, had the features previously associated with obstructed detrusor, or detrusor with impaired contractility. CONCLUSIONS: The trigonal muscle does not undergo structural changes as previously described in the detrusor in association with voiding dysfunction. Its contraction depends on electrical coupling of its muscle cells, and has a supportive role in normal micturition, mooring the terminal ureters to the bladder base, to allow efflux and guard against reflux of urine. Activation of volume and tension sensory neuroterminals may contribute to some storage and voiding micturition reflexes, and may be related to normal and abnormal perception of urge.

Aged↗

Noninflammatory chronic pelvic pain syndrome: immunological study in blood, ejaculate and prostate tissue.

OBJECTIVES: The aim of this prospective study was to observe immunophenotypic patterns in patients with noninflammatory chronic pelvic pain syndrome (Cat IIIB CPPS) for further description and as possible surrogate markers for diagnosis and treatment. METHODS: Eighty-eight patients with a referral diagnosis of chronic prostatitis underwent fractionated urinary cultures including expressed prostate secretion (EPS) and ejaculate analysis twice on two occasions. Monthly serum analyses included C3c, C4, IL-1alpha, sIL-2R, and IL-6. One hundred samples from healthy individuals were used as the control group for serum analysis. Monthly ejaculate testing was done 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 men without symptoms or obstruction. RESULTS: Complete sampling of urinary, serum and ejaculate specimens was achieved in 50/88 (57%) patients. Cat IIIB CPPS was observed in 44/50 (88%) patients. Intra-acinar T-lymphocytic infiltrates were dominated by T cytotoxic cells (p = 0.05). Immunohistochemical studies showed inflammatory expression in serum complement, serum interleukin, and ejaculate interleukin concentrations in relation to the presence of large numbers of T cells (all p values < or =0.01). No difference was found in the proportion of B lymphocytes in patients with Cat IIIB CPPS compared to the control group. Serum and ejaculate IL-6 and ejaculate IgA increased significantly and dropped again, correlating with a release of clinical symptoms. CONCLUSIONS: Interleukin, complement and immunoglobulin determinations in serum and ejaculate reveal an inflammatory process even in Cat IIIB CPPS. The findings of intra-acinar T-cell-rich infiltrates and the associated inflammatory reaction may be a significant advance in defining Cat IIIB CPPS caused by a possible autoimmune component. Serum and ejaculate IL-6 and ejaculate IgA are possible surrogate markers for the diagnosis and treatment of Cat IIIB CPPS.

Adult↗

Purification and characterization of the first archaeal aconitase from the thermoacidophilic Sulfolobus acidocaldarius.

The first archaeal aconitase was isolated from the cytosol of the thermoacidophilic Sulfolobus acidocaldarius. Interestingly, the enzyme was copurified with an isocitrate lyase. This enzyme, directly converting isocitrate, the reaction product of the aconitase reaction, was also unknown in crenarchaeota, thus far. Both proteins could only be separated by SDS gel electrophoresis yielding apparent molecular masses of 96 kDa for the aconitase and 46 kDa for the isocitrate lyase. Despite of its high oxygen sensitivity, the aconitase could be enriched 27-fold to a specific activity of approximately 55 micromol x min(-1) x mg(-1), based on the direct aconitase assay system. Maximal enzyme activities were measured at pH 7.4 and the temperature optimum for the archaeal enzyme was recorded at 75 degrees C, slightly under the growth optimum of S. acidocaldarius around 80 degrees C. Thermal inactivation studies of the aconitase revealed the enzymatic activity to be uninfluenced after one hour incubation at 80 degrees C. Even at 95 degrees C, a half-life of approximately 14 min was determined, clearly defining it as a thermostable protein. The apparent K(m) values for the three substrates cis-aconitate, citrate and isocitrate were found as 108 microM, 2.9 mM and 370 microM, respectively. The aconitase reaction was inhibited by the typical inhibitors fluorocitrate, trans-aconitate and tricarballylate. Amino-acid sequencing of three internal peptides of the S. acidocaldarius aconitase revealed the presence of highly conserved residues in the archaeal enzyme. By amino-acid sequence alignments, the S. acidocaldarius sequence was found to be highly homologous to either other putative archaeal or known eukaryal and bacterial sequences. As shown by EPR-spectroscopy, the enzyme hosts an interconvertible [3Fe--4S] cluster.

Aconitate Hydratase↗

Posttranslationally processed forms of the human chemokine HCC-1.

HCC-1 is the only CC-chemokine known so far which circulates in nanomolar concentrations in human plasma. Its physiological function is not well defined. Posttranslational processing of HCC-1 was shown to modulate its biological properties. In this study several different processed forms of HCC-1 were isolated. Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. These peptides were isolated from a peptide library of human blood filtrate and represent predominantly HCC-1 (1-74) and glycosylated HCC-1 (1-74). Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. Furthermore N-terminally truncated HCC-1 (3-74) and HCC-1 (4-74) were identified in the peptide library. In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). Our data imply that HCC-1 (1-74), HCC-1 (3-74), HCC-1 (4-74) and glycosylated HCC-1 (1-74) circulate in human blood. The N-terminal processing and modification of HCC-1 might be of importance in displaying its full biological activity.

Amino Acid Sequence↗

Seminal vesicle-sparing radical prostatectomy: a novel concept to restore early urinary continence.

OBJECTIVES: Urinary incontinence after radical prostatectomy continues to be a distressing problem, even with preservation of the neurovascular bundles and meticulous apical dissection. Recent studies suggest that motor and sensory components of the pelvic nerve may be affected by surgery, since both components are anatomically located in intimate contact with the seminal vesicles. We propose seminal vesicle-sparing radical prostatectomy to preserve pelvic innervation and improve the rate of urinary continence. METHODS: Fifty-four patients were enrolled in this prospective study. A standard retropubic radical prostatectomy was performed in 34 patients. A seminal vesicle-sparing radical prostatectomy was performed in a pilot series of 20 consecutive patients. The seminal vesicle tip and surrounding tissue were preserved and carefully handled. In all patients, a modified pad test and posterior urethral sensory threshold test were performed preoperatively and 6 weeks and 6 months postoperatively and correlated with urinary continence. RESULTS: The intraoperative preservation of the seminal vesicle tip was possible in all patients in this pilot series (n = 20). In the seminal vesicle-sparing radical prostatectomy group, the continence rate was 60% after 6 weeks and 95% after 6 months. These rates were significantly higher than the continence rates in the standard prostatectomy group (18% and 82% at 6 weeks and 6 months, respectively). The sensory threshold levels in the seminal vesicle-sparing group were similar to the preoperative values and were significantly lower than the postoperative threshold levels in the standard prostatectomy group. CONCLUSIONS: Seminal vesicle tip-sparing radical prostatectomy may be a surgical option to preserve pelvic innervation and maintain urinary continence after radical prostatectomy. Further randomized studies are necessary to elucidate the impact of seminal vesicle-sparing radical prostatectomy on restoration of urinary continence.

Aged↗

Early orchiopexy: prepubertal intratubular germ cell neoplasia and fertility outcome.

OBJECTIVES: To investigate the prepubertal prevalence of intratubular germ cell neoplasia of the unclassified type (ITGCNU) and its significance as a predictor of testicular cancer and to evaluate the effect of early orchiopexy (at younger than 2 years of age) on subsequent fertility of patients with bilateral cryptorchidism. METHODS: Testicular biopsies (n = 660) from 440 prepubertal patients with cryptorchidism who underwent orchiopexy between January 1, 1970 and December 31, 1979 were evaluated for ITGCNU using placental-like alkaline phosphatase (PLAP) antibody. The clinical outcome in 15 patients with PLAP-positive germ cells was evaluated in 1997. In addition, the effect of age at surgery on the fertility of patients with bilateral cryptorchidism was assessed by clinical follow-up until 1997 and was correlated with the histologic data at orchiopexy. RESULTS: PLAP-positive germ cells morphologically identical with adult ITGCNU were found in the biopsies of 22 patients (5%). After more than two decades, none of the 15 patients with successful follow-up developed testicular cancer. The fertility outcome in the patients with bilateral cryptorchidism correlated with the number of spermatogonia at orchiopexy (P = 0.018), but correlated inversely with age at orchiopexy (P = 0.021). CONCLUSIONS: PLAP-positive germ cells in prepubertal testicular biopsy specimens are not necessarily precursors of testicular cancer after orchiopexy. In addition, our data support the idea that early orchiopexy may be beneficial in preventing infertility.

Adolescent↗

Kallmann's syndrome: clues to clinical diagnosis.

Hypogonadotropic patients may visit pediatricians, general practitioners, endocrinologists or urologists, presenting with microphallus, cryptochidism or pubertas tarda and delayed bone maturation. Congenital hypogonadotropic hypogonadism is characterized, apart from small testes, by the constellation of low serum levels of testosterone, LH and FSH. Kallman's syndrome is characterized by congenital hypogonadotropic hypogonadism with midline defects such as anosmia (a deficiency of the sense of smell). The first case report dates back to 1856, and genetic defects causing the syndrome have been recently described. The diagnosis can be clinically suspected and is established by confirming hormonal studies.

Adolescent↗

Kallmann's syndrome: clues to clinical diagnosis.

Five cases of Kallmann's syndrome are presented, out-patients with microtestes, hypogonadotropic hypogonadism and complete anosmia. The final diagnosis was made only when they were aged between 17 and 26 (mean 21 years), although they had been seen by several physicians before: 3 for cryptorchidism and 3 for absence of spontaneous puberty; 2 had a positive family history, and 4 of the 5 patients or their parents admitted that they were aware of the fact that their sense of smell was completely absent, but they did not mention it spontaneously.

Adolescent↗

Effect of radical prostatectomy on sensory threshold and pressure transmission.

PURPOSE: The mechanisms involved in post-radical prostatectomy incontinence remain unclear despite previous anatomical and functional studies. In addition, the factors responsible for the restoration of continence are not well studied. To improve our understanding of the alterations in continence mechanisms, we prospectively investigated the temporal changes in urodynamic parameters after radical prostatectomy. MATERIALS AND METHODS: Cystometry, urethral pressure profilometry and posterior urethral sensory threshold measurements were performed in patients undergoing radical prostatectomy. Preoperative pressure transmission was determined by the maximal urethral pressure divided by the maximal abdominal pressure during cough maneuvers at a bladder volume of 200 ml. Postoperative sensory threshold, pressure transmission (% of pressure transmission), maximal urethral closure pressure and functional sphincter length were measured 6 weeks and 6 months after prostatectomy. These parameters were compared between continent and incontinent patients. RESULTS: Preoperative and postoperative urodynamic studies were completed in 34 patients. The continence rate after 6 weeks was 18% (6 patients) and improved to 82% (28) after 6 months. Preoperatively sensory threshold was 16 +/- 11 mA. After 6 weeks and 6 months, respectively, sensory threshold was significantly higher in incontinent (84 +/- 11 mA., 70 +/- 8 mA.) compared to continent (65 +/- 8 mA., 41 +/- 12 mA.) patients. Preoperative proximal urethral sensory threshold was not correlated with degree of postoperative incontinence determined by pad tests. Pressure transmission was not different in continent and incontinent patients postoperatively. After 6 weeks and 6 months, respectively, pressure transmission was 77% and 91% in continent, and 37% and 58% in incontinent patients (p = 0.04, p = 0.03). Maximal urethral closure pressure was significantly higher in continent patients (35 +/- 6 cm. H2O) compared to incontinent patients (11 +/- 9 cm. H2O). Sphincter length decreased from 50 mm. preoperatively to 24 mm. after 6 weeks and 25 mm. after 6 months. There was no difference in sphincter length between continent and incontinent patients. CONCLUSIONS: Posterior urethral sensitivity and pressure transmission are impaired immediately after prostatectomy. An improvement in these parameters after 6 months is associated with the restoration of continence. These observations suggest that urinary continence after radical prostatectomy depends on the integrity of posterior urethral sensation and the efficiency of pressure transmission.

Humans↗

Novel glycosylated forms of human plasma endostatin and circulating endostatin-related fragments of collagen XV.

Circulating elongated forms of the angiogenesis inhibitor and potential anti-cancer drug endostatin were isolated from human blood filtrate. Immunoreactive endostatin was identified by a polyclonal rabbit antiserum raised against an N-terminal epitope of the polypeptide and purified by consecutive chromatographic steps and immunoblotting. N- and C-terminal sequence analyses of the isolated molecules revealed different forms of endostatin starting with V(117)HLRPAR. lacking the last and final three residues of the noncollagenous domain 1 (NC-1) of collagen XVIII, respectively. These polypetides are found to be O-glycosylated at T(125) (residue 9) with a glycan structure of the mucin type consisting of galactose N-acetylgalactosamine and N-acetylneuraminic acid residues. Carbohydrate analyses were performed via the semiquantitative HPLC-electrospray ionization mass spectrometry (ESMS) technique after exoglycosidase hydrolysis. Circulating endostatins are present as sialoglycoprotein (22 000 and 21 841 Da +/- 0.02%) and asialoglycoprotein structures (21 710 and 21 549 Da +/- 0.02%), while the two completely deglycosylated forms are obtained only after enzymatic incubation. The described glycosylated endostatins may represent intermediates in the proteolytic pathway of the NC-1 domain of collagen XVIII resulting in bioactive endostatins. Furthermore, immunoreactive endostatin-related C-terminal fragments of human collagen XV are found in the hemofiltrate. These polypeptides exhibit the N-terminal sequences P(66)HLLPPP. and Y(81)EKPALH. of the collagen XV NC-1 domain. ESMS and immunoblotting analyses reveal three glycosylated polypeptides with a molecular mass ranging from 16 to 21 kDa. Due to the high degree of homology between collagen XV and collagen XVIII as well as their analoqous proteolytic processing, functional similarities of collagen XVIII- and XV-related fragments should be revealed in future experiments.

Amino Acid Sequence↗

[Ureteral stent placement in hydronephrosis during pregnancy].

In this study we describe 22 cases of retrograde ureteral stent placement in pregnant women with therapy-resistant flank pain due to hydronephrosis. Eleven were primiparous and one patient expected twins. Eight of 22 patients presented symptoms of pyelonephritis. In 21 cases the hydronephrosis was located on the right and in 4 cases it was bilateral. Maximal lower calix diameter was 12 mm (range 9-22 mm). With the exception of two cases, sonographically controlled stent placement was performed under local anesthesia without sedation. All patients were painfree within 6 days and were given prophylactic low doses of antibiotic until the day of delivery. Renal function remained within the normal limits. Double-J stent displacement occurred in 3 patients - of which one underwent nephrostomy. Postnatal examination demonstrated urolithiasis in 4 of 19 patients. This study provides evidence for effectiveness of retrograde ureteral Double-J stent placement as a therapeutic option in cases of severe symptomatic hydronephrosis during pregnancy with a low morbidity rate.

Adult↗

Determination of the pK(a) of 12-S-hydroxyheptadecatrienoic acid by studies of its vial surface adsorption using high-performance liquid chromatography and liquid scintillation technique.

When dissolving 12-S-hydroxyheptadecatrienoic acid (HHT) in aqueous buffer systems, a partition equilibrium is achieved between the liquid buffer phase and the solid vial phase. This adsorption equilibrium is dependent on the vial material, the ratio of solvent volume to moistened vial surface area (V/A), the pH of the aqueous solution the buffer additives and the contact time. Partition ratios (E) were established following determination of the recoveries (omega) of HHT in the liquid phases. The pK(a) of HHT could be calculated to be 4.4 +/- 0.2. The HHT concentration in the liquid phase rose considerably when increasing the pH value (17% at pH 1.9, up to 97% of applied HHT at pH 8.0) and increasing the V/A-ratio, as well as when using glass instead of polypropylene (PP) or adding acetonitrile as an organic solvent and HHT-affine substances like albumins or globulin. The HHT concentration was found to decrease during contact to PP surfaces as a result of HHT adsorption and diffusion into the hydrophobic vial material. This effect could be minimized by adding a larger percentage of acetonitrile (50-60%).

Adsorption↗

Pheochromocytomas: can malignant potential be predicted?

OBJECTIVES: The presence of metastatic lesions is the only acceptable fact to confirm malignant pheochromocytoma. Patients with malignant pheochromocytomas, however, have a very poor survival rate. The aim of our study was to postulate predictive values for malignant pheochromocytomas. METHODS: We evaluated symptoms, diagnostic modalities, treatment, and long-term follow-up of 86 patients with 85 benign and 10 malignant pheochromocytomas. Parameters from the benign were compared with those of the malignant pheochromocytomas. RESULTS: Preoperative 24-hour urinary dopamine was in the normal range for benign pheochromocytomas but increased in malignant pheochromocytomas (P<0.0001). Vanillylmandelic acid was elevated in both benign and malignant pheochromocytomas but higher in malignant than in benign tumors (P = 0.01). No differences could be shown in urinary epinephrine and norepinephrine samplings. Tumor location was divided into 77 adrenal (81%) and 18 extra-adrenal (19%) sites. Malignant pheochromocytomas were located more often at extra-adrenal sites (P = 0.03). There was no increased incidence of malignancy in patients with familial bilateral pheochromocytomas or multiple endocrine neoplasia. Tumors greater than 80 g in weight corresponded to malignancy (P<0.0001). Dopamine tumor concentration was higher in malignant than in benign pheochromocytomas (P = 0.01). Persistent arterial hypertension occurred in 9 (13%) of 72 benign and 6 (60%) of 10 malignant pheochromocytomas (P = 0.001). The 10-year survival rate was 94% for benign pheochromocytomas. All patients with malignant pheochromocytomas died within this period (P = 0.0001). CONCLUSIONS: High preoperative 24-hour urinary dopamine levels, extra-adrenal tumor location, high tumor weight, elevated tumor dopamine concentration, and postoperative persistent arterial hypertension are all factors that increase the likelihood of malignant pheochromocytoma. Patients with these characteristics should have more frequent follow-up evaluations to identify malignancy at earlier states.

Adolescent↗

Variable response to intracavernous prostaglandin E1 testing for erectile dysfunction.

OBJECTIVES: Anxiety and apprehension may negatively influence the erectile response to the first intracavernous injection with vasoactive agents. This may result in too many false-positive diagnoses of vascular insufficiency if the first injection ever made in a patient is used for color Doppler duplex sonography (CDS) evaluation. METHODS: One hundred sixty-eight consecutive patients (aged 18 to 75 years) with erectile dysfunction underwent a standardized evaluation, including the intracavernous injection test (ICIT) stimulated with 10 microg prostaglandin E1. Responses were recorded on a four-point scale: no response = 0, tumescence = 1, rigidity sufficient for intercourse = 2, full erection = 3. ICIT was repeated after 10 days and combined with CDS. The clinically assessed response to ICIT was correlated with end-diastolic flow velocity. RESULTS: Of 168 patients, 114 (68%) responded equally to the first and second ICIT, but 45 (27%) had an improved response, from tumescence to full erection in the second test (P <0.0001); in 9 (5%), the response deteriorated. The overall mean response was 1.6 (95% confidence interval 1.5 to 1.7) and 1.9 (95% confidence interval 1.7 to 2.0) (P <0.0001) for the first and second test, respectively. Of 168 patients, 89 (53%) responded with erections sufficient for intercourse when tested the first time and 104 (62%) did so after the second injection. CONCLUSIONS: Erectile response to diagnostic intracavernous injection of prostaglandin E1 significantly improved in the second compared with the first test. Therefore, cautious interpretation of CDS is advised when patients are injected for the first time because too many false-positive tests may result.

Adolescent↗

[Kidney and urinary tract diseases in pregnancy].

Management of urologic disorders in pregnant patients often increases the anxiety of all involved. Based on a thorough understanding of the physiologic changes seen in various organ systems the urologist has to assume the responsibility for the well-being of the mother and the fetus. Apart from the urinary tract infection, which occurs as frequent as in non-pregnant patients but has a significantly higher risk of acute bacterial pyelonephritis, it is mainly the pregnancy-associated symptomatic hydronephrosis and the urolithiasis which are complicating approximately 1 of every 1000-1500 pregnancies. Urinary tract infections should be treated in any case by antibiotics according to a antibiogram. High risk patients with history of vesicoureteral reflux or recurrent pyelonephritis should be treated prophylactically. Following parturition these patients should be investigated urologically to exclude structural abnormalities of the genitourinary system. In case of symptomatic hydronephrosis and calculous disease the first approach should be a watchful conservatism with symptomatic relief. If the symptoms persist insertion of a double-J-stent or in case of live-threatening situations (e.g. urosepsis) when urgent decompression and rapid evacuation is mandatory a percutaneous nephrostomy can be brought in place under sonographic monitoring completely thereby avoiding any radiation exposure.

Adult↗

Clinical value of pelvic and penile magnetic resonance angiography in preoperative evaluation of penile revascularization.

Penile angiography is invasive, costly and requires postinterventional surveillance. The aim of this pilot study was to determine whether three dimensional magnetic resonance (3D-MR)-angiography may replace conventional penile angiography in preoperative planning of penile revascularization. Twelve patients with a mean age of 39 (21-59) y were evaluated. All patients underwent evaluation with intracavernous pharmacotesting, color Doppler sonography (CDS), digital subtraction angiography (DSA) and pelvic MR-angiography with gadolinium diethylene-triaminepentaacetic acid (Gd-Dota) 0.2-0.3 mmol/kg body weight. MR-angiography demonstrated the anatomy of the internal iliac arteries in 9 out of 12 patients. Intrapenile vessels were visible in 7 out of 12 patients. In comparison DSA provided complete visualization of all pelvic and penile vessels. Relevant arterial obstruction was found in 10 out of 12 patients. CDS revealed a mean maximal arterial flow of 27 (22-40) cm/s and showed in accordance to angiography arterial insuffiency in 10 out of 12 cases. Indication for revascularization could have been based on MR-angiography alone in only one patient. Therefore, selective penile angiography remains the 'gold standard' for preoperative planning of revascularization procedures.

Adult↗