In vitro formation of the anthranoid scaffold by cell-free extracts from yeast-extract-treated Cassia bicapsularis cell cultures.

The anthranoid skeleton is believed to be formed by octaketide synthase (OKS), a member of the type III polyketide synthase (PKS) superfamily. Recombinant OKSs catalyze stepwise condensation of eight acetyl units to form a linear octaketide intermediate which, however, is incorrectly folded and cyclized to give the shunt products SEK4 and SEK4b. Here we report in vitro formation of the anthranoid scaffold by cell-free extracts from yeast-extract-treated Cassia bicapsularis cell cultures. Unlike field- and in vitro-grown shoots which accumulate anthraquinones, cell cultures mainly contained tetrahydroanthracenes, formation of which was increased 2.5-fold by the addition of yeast extract. The elicitor-stimulated accumulation of tetrahydroanthracenes was preceded by an approx. 35-fold increase in OKS activity. Incubation of cell-free extracts from yeast-extract-treated cell cultures with acetyl-CoA and [2-(14)C]malonyl-CoA led to formation of torosachrysone (tetrahydroanthracene) and emodin anthrone, beside two yet unidentified products. No product formation occurred in the absence of acetyl-CoA as starter substrate. To confirm the identities of the enzymatic products, cell-free extracts were incubated with acetyl-CoA and [U-(13)C(3)]malonyl-CoA and (13)C incorporation was analyzed by ESI-MS/MS. Detection of anthranoid biosynthesis in cell-free extracts indicates in vitro cooperation of OKS with a yet unidentified factor or enzyme for octaketide cyclization.

The correlation between prostate volume, transition zone volume, transition zone index and clinical and urodynamic investigations in patients with lower urinary tract symptoms.

OBJECTIVE: To determine if, in patients with lower urinary tract symptoms (LUTS), measurement of the transition zone (TZ) of the prostate by transrectal ultrasonography (TRUS) and the ratio between the TZ volume and total prostate volume (TZ index) correlates better with clinical and urodynamic investigations than total prostate volume alone. PATIENTS AND METHODS: In total, 150 consecutive patients with LUTS underwent a standardized screening programme including the International Prostate Symptom Sore (IPSS), a physical examination, TRUS of the prostate and urodynamic investigations with pressure-flow studies. The total prostate volume and TZ volume were assessed from TRUS using the ellipsoid formula. Spearman's rank correlation coefficients were calculated between different prostate volume measurements and specific symptomatic and urodynamic variables. RESULTS: The relationships between specific IPSS symptoms, symptom scores and the prostate volume measurements were not statistically significant except for one domain, nocturia, that appeared to be statistically significantly correlated with the TZ index (r = 0.25). The correlations for free flow, pressure-flow variables and prostate volume measurements were stronger, but only moderate at best. The highest correlations were between TZ volume and the linear passive urethral resistance obstruction category, urethral resistance factor and detrusor pressure at maximum flow (r = 0.43, 0.44 and 0.40, respectively). The differences between the correlations of prostate volume and TZ index and these variables were small (r = 0.39, 0.38 and 0.37, respectively for prostate volume and r = 0.38, 0.40 and 0.33 respectively for TZ index). CONCLUSIONS: There were very small differences between the correlations of total prostate volume, TZ volume and TZ index, and clinical and pressure-flow variables. In the assessment of the last two, the estimation of the total prostate volume by TRUS was a reasonable way to obtain the required information about prostate size and measuring TZ volume and calculating TZ index was of limited additional value. Symptoms and bladder outlet obstruction were mainly determined by other factors than the prostate and, specifically, TZ volume. As earlier studies have indicated that including pressure-flow data in the pre-operative evaluation and selection of patients for interventional therapies may improve the overall clinical results, we think that prostate volume, TZ volume or symptoms alone should not be used as the main indication for deciding on the appropriate invasive treatment options.

The value of antibiotic prophylaxis during extracorporeal shock wave lithotripsy in the prevention of urinary tract infections in patients with urine proven sterile prior to treatment.

INTRODUCTION: There are controversies in the literature regarding the need for and duration of antibiotic prophylaxis in patients treated with extracorporeal shock wave lithotripsy (ESWL) who have a negative urine culture before treatment. In order to determine the efficacy of antibiotic prophylaxis in ESWL treatment of patients with proven sterile urine, a randomized trial was performed. METHODS: Patients were randomized for placebo and 1 or 7 days antibiotic prophylaxis (cefuroxime or ciprofloxacin), starting 30 min before ESWL. Post-ESWL studies (immediately and 2 and 6 weeks after ESWL) included patient history, urine culture and Gram stain. RESULTS: After 2 weeks 20% of the patients and after 6 weeks 23% of the patients had bacteriuria, but there was no statistical significance between patients treated with placebo or those receiving prophylactic treatment. Only 2-3% of the patients (in the prophylaxis and placebo group) had clinical and bacteriological signs of a urinary tract infection, either 2 or 6 weeks after ESWL, possibly caused by re-infection, however, since bacteria were found in none of the urine samples collected directly after ESWL. There was no beneficial effect of antibiotic prophylaxis, in the prevention of urinary tract infections in patients with a nephrostomy catheter or dilatation at the site of treatment. CONCLUSION: We conclude that in patients with urine proven sterile prior to ESWL there is no need for antibiotic prophylaxis.

Correlation between uroflowmetry, prostate volume, postvoid residue, and lower urinary tract symptoms as measured by the International Prostate Symptom Score.

OBJECTIVES: To evaluate the relation between lower urinary tract symptoms (LUTS) as measured by the International Prostate Symptom Score (IPSS) and noninvasive objective parameters of lower urinary tract dysfunction. METHODS: Eight hundred three consecutive patients with LUTS and/or benign prostatic hyperplasia were evaluated with IPSS, uroflowmetry, prostate volume estimation, and postvoiding residue measurement. The relations between these parameters were quantified by means of Spearman correlation coefficients. RESULTS: Statistically significant but weak correlations were found between the IPSS and results of uroflowmetry and postvoiding residual urine. There was no correlation between the IPSS and results of prostate volume measurements. CONCLUSIONS: The correlation between objective noninvasive parameters of lower urinary tract dysfunction and LUTS is weak.

The outcome of renal ultrasound in the assessment of 556 consecutive patients with benign prostatic hyperplasia.

PURPOSE: We determined the additional value of renal ultrasonography in the assessment of patients with benign prostatic hyperplasia. MATERIALS AND METHODS: Renal ultrasound was performed in 556 consecutive patients with benign prostatic hyperplasia and the results were correlated with other clinical parameters. RESULTS: Of the patients 14 (2.5%) had dilatation of the renal pelvis, 65 (11.7%) had renal cysts and 1 (0.18%) had renal cell carcinoma. The serum creatinine level appeared to be correlated with dilatation of the renal pelvis. To predict dilatation additional information can be obtained by including the results of the post-void residual measurements. CONCLUSIONS: Renal ultrasound is indicated only in patients with a specific serum creatinine level and/or post-void residual volume.

The predictive value of microscopic haematuria in patients with lower urinary tract symptoms and benign prostatic hyperplasia.

OBJECTIVE: To determine the additional value of the presence of microscopic haematuria in patients with benign prostatic hyperplasia (BPH). METHODS: In 750 consecutive patients with BPH urinalysis was performed and the grade of microhaematuria was correlated with other clinical findings. RESULTS: Microscopic haematuria was found in one third of the patients. Only 3 had a bladder tumour and 49 patients had urinary calculi for which only one patient required treatment. There was no correlations between any clinical parameter and the finding of microscopic haematuria. CONCLUSION: Microscopic haematuria is a frequent finding in assessment of BPH patients and additional tests should only be performed if indicated.