The long-term learning curve of holmium laser enucleation of the prostate (HoLEP) in the en-bloc technique: a single surgeon series of 500 consecutive cases.

PURPOSE: To evaluate perioperative parameters, clinical outcomes, and the learning curve of holmium laser enucleation of the prostate (HoLEP) of a single surgeon in 500 consecutive cases. METHODS: Demographic parameters, outcomes, and adverse events were evaluated. The learning curve for HoLEP in en-bloc technique of the first 500 consecutive patients was analyzed in clusters of 100 (clusters 1-5) using the Wilcoxen rank test, Chi² test and Kruskal Wallis test. RESULTS: Enucleation weight was similar in the clusters 1,2,3, and 5 (62 g, 63 g, 61 g, 61 g), in cluster 4 it was slightly higher at 73 g. There was a significant reduction in operating time from 67 min (cluster 1) to 57 min (cluster 2), 46 min (cluster 3), 53 min (cluster 4), and 43 min (cluster 5), p < 0.001. Enucleation efficiency (g/min) showed a steady increase (1.72, 2.24, 2.79, 2.92 vs. 2.99, p < 0.001). Laser energy efficiency also improved (2.17 vs. 2.12 vs. 1.71 vs. 1.65 vs. 1.55; p < 0.001). There was no measurable learning curve regarding the length of hospital stay (mean 2.5 days), catheterization time (1.9 days), hemoglobin drop (approx. 1 g/dl) or complications (p > 0.1). CONCLUSIONS: HoLEP using the en-bloc technique is a safe and highly effective method. Over time, a slight but steady learning curve and improvement in operation time, enucleation efficiency and laser energy efficiency were shown even for an experienced surgeon - after 500 cases, still no plateau was reached. There was no measurable learning curve regarding blood loss, complications, length of hospital stay, and catheterization time.

Comparison of endoscopic versus CT assessment of stone-free status after percutaneous nephrolithotomy (PCNL).

This study is aimed to determine whether postoperative low dose computed tomography (LDCT) imaging is necessary after percutaneous nephrolithotomy (PCNL), or the surgeon's intraoperative assessment of residual fragments (RF) is sufficient and avoidance of postoperative imaging with reduction of radiation exposure can be achieved. Data of all 610 patients who underwent PCNL in prone position in our institution from February 2009 to September 2020 was collected. Parameters such as age, gender, BMI, ASA-Classification, stone related parameters and the surgeon's assessment of stone-free status were analyzed. The LDCT performed postoperatively was compared to the intraoperative assessment of the surgeon regarding RF. The mean age of patients was 52.82 years; the mean BMI was 28.18 kg/m2. In 418 cases, the surgeon made a clear statement about the presence of RF and postoperative LDCT was carried out. The discrepancy between the two methods (surgeon´s assessment vs. LDCT) was significant at p < 0.0001. The sensitivity, specificity, positive and negative predictive value of the surgeon when assessing RF were 24.05%, 99.45%, 98.28% and 50%. Stone free rate (SFR) after primary PCNL was 45.57%. The overall SFR at discharge was 96.23%. Although the surgeon´s assessment of RF was reliable, postoperative LDCT imaging should still be performed if endoscopic stone clearance is suspected due to the high false negative rate and the low negative predictive value. The optimal timing of postoperative imaging following PCNL remains unclear.

The role of AtMUS81 in DNA repair and its genetic interaction with the helicase AtRecQ4A.

The endonuclease MUS81 has been shown in a variety of organisms to be involved in DNA repair in mitotic and meiotic cells. Homologues of the MUS81 gene exist in the genomes of all eukaryotes, pointing to a conserved role of the protein. However, the biological role of MUS81 varies between different eukaryotes. For example, while loss of the gene results in strongly impaired fertility in Saccharomyces cerevisiae and nearly complete sterility in Schizosaccharomyces pombe, it is not essential for meiosis in mammals. We identified a functional homologue (AtMUS81/At4g30870) in the genome of Arabidopsis thaliana and isolated a full-length cDNA of this gene. Analysing two independent T-DNA insertion lines of AtMUS81, we found that they are sensitive to the mutagens MMS and MMC. Both mutants have a deficiency in homologous recombination in somatic cells but only after induction by genotoxic stress. In contrast to yeast, no meiotic defect of AtMUS81 mutants was detectable and the mutants are viable. Crosses with a hyperrecombinogenic mutant of the AtRecQ4A helicase resulted in synthetic lethality in the double mutant. Thus, the nuclease AtMUS81 and the helicase AtRecQ4A seem to be involved in two alternative pathways of resolution of replicative DNA structures in somatic cells.

Comparison of the contact surface pattern between virtual and milled Cerec 3D full-ceramic crowns.

PURPOSE: The objective of this study was to compare resulting contact point patterns in the context of crown production with the Cerec 3D system. The contact surface patterns of full-ceramic crowns milled with the Cerec 3D system were examined for agreement with the corresponding virtually produced crowns. MATERIALS AND METHODS: The agreement was determined with regard to the parameters number, size, position, and shape. Twenty-eight models were mounted centrally in a mechanical articulator. In these model cases, at least one second premolar and one first molar in the maxilla and mandible had to be prepared for crown production (preparation). A total of 50 teeth was prepared. Crowns were then designed with the aid of the Cerec 3D system. The crown designs were milled and integrated in the model. Silicone bite registrations, the contact point patterns of which were compared with the virtually designed crowns, were produced in static occlusion in the region of the crowns. Different evaluation methods were used for the comparison. RESULTS: There was an agreement of 78% with regard to number, 76% in terms of localization, 65% regarding size, and 65% in terms of shape. CONCLUSION: It was thus confirmed that very good, highly reproducible results can be achieved with the Cerec 3D system.

Molecular characterization of homologues of both subunits A (SPO11) and B of the archaebacterial topoisomerase 6 in plants.

The Spo11 protein is an eukaryotic homologue of the topoisomerase 6 subunit A from archaebacteria. In yeast Spo11p has been found to bind covalently to double-strand breaks (DSBs) during meiosis. Single homologues of the SPO11 gene exist in various eukaryotes, except plants. Previously, we found in the Arabidopsis thaliana genome two ancient paralogs, AtSPO11-1 and 2. Here we report on the molecular characterization of a third one, AtSPO11-3. This puzzling finding might be explained by the fact that we detected additionally--for the first time outside of the archaebacterial kingdom--a homologue of the subunit B of topoisomerase 6, AtTOP6B. Both AtSPO11-3 and AtTOP6B are abundantly expressed in Arabidopsis and EST comparisons indicate the presence of both genes in various plant species. Via two hybrid studies we could demonstrate that full length AtTop6B is able to interact with AtSpo11-2 and 3 but not with AtSpo11-1. Our data suggest that plants possess in contrast to other eukaryotes an additional archaebacterial kind of topoisomerase.

Molecular characterisation of RecQ homologues in Arabidopsis thaliana.

Members of the RecQ family of DNA helicases are involved in processes linked to DNA replication, DNA recombination and gene silencing. RecQ homologues of various animals have been described recently. Here, for the first time for plants, we characterised cDNAs of all in all six different RecQ-like proteins that are expressed to different extents in Arabidopsis thaliana. Surprisingly, three of these proteins are small in size [AtRecQl1, AtRecQl2, AtRecQl3-606, 705 and 713 amino acids (aa), respectively], whereas the two bigger proteins result from a duplication event during plant evolution [AtRecQl4A and AtRecQl4B-1150 and 1182 aa, respectively]. Another homologue (AtRecQsim, 858 aa) most probably arose by insertion of an unrelated sequence within its helicase domain. The presence of these homologues demonstrates the conservation of RecQ family functions in higher eukaryotes. We also detected a small gene (AtWRNexo) encoding 285 aa which, being devoid of any RecQ-like helicase domain, reveals a striking homology to the exonuclease domain of human Werner protein, a prominent RecQ helicase of larger size. By means of the two-hybrid assay we were able to detect an interaction between AtWRNexo and AtRecQl2, indicating that activities that reside in a single protein chain in mammals might in plants be complemented in trans.

Molecular characterisation of two paralogous SPO11 homologues in Arabidopsis thaliana.

The Spo11 protein of yeast has been found to be covalently bound to double-strand breaks in meiosis, demonstrating a unique role of the protein in the formation of these breaks. Homologues of the SPO11 gene have been found in various eukaryotes, indicating that the machinery involved in meiotic recombination is conserved in eukaryotes. Here we report on SPO11 homologues in plants. In contrast to what is known from other eukaryotes, Arabidopsis thaliana carries in its genome at least two SPO11 homologues, AtSPO11-1 and AtSPO11-2. Both genes are not more closely related to each other than to other eukaryotic SPO11 homologues, indicating that they did not arise via a recent duplication event during higher plant evolution. For both genes three different poly-adenylation sites were found. AtSPO11-1 is expressed not only in generative but also to a lesser extent in somatic tissues. We were able to detect in different organs various AtSPO11-1 cDNAs in which introns were differently spliced-a surprising phenomenon also reported for SPO11 homologues in mammals. In the case of AtSPO11-2 we found that the 3' end of the mRNA is overlapping with a mRNA produced by a gene located in inverse orientation next to it. This points to a possible antisense regulation mechanism. Our findings hint to the intriguing possibility that, at least for plants, Spo11-like proteins might have more and possibly other biological functions than originally anticipated for yeast.

Investigations on the use of C-21-steroids as precursors for placental oestrogen synthesis in the cow.

Cotyledonary homogenates from 220 and 270 day pregnant and term cows were incubated (NADPH- and NAD(+)-regenerating system) with 3H-pregnenolone and 3H-progesterone, respectively. Identification of metabolites was based on separation on HPLC and the respective retention times. On days 220/270 the major metabolite formed after incubation with 3H-pregnenolone was progesterone, followed by 17 alpha-hydroxyprogesterone and dehydroepiandrosterone/17 alpha-hydroxypregnenolone; the formation of estrone was low (up to 6%), while it was the major metabolite after incubation of pregnenolone with term placenta. At all stages of pregnancy investigated, the only metabolite found after incubation with 3H-progesterone was 17 alpha-hydroxyprogesterone. These data provide evidence that 17 alpha-hydroxyprogesterone is the endpoint of steroid biosynthesis in the bovine placenta along the delta 4-route and that oestrogen synthesis follows the delta 5-pathway. Based on the high activity of 3 beta-hydroxysteroiddehydrogenase/delta 5/4-isomerase also found on days 220/270, the key enzyme allowing for placental oestrogen synthesis in the cow seems to be cytochrome P450c17 alpha. Thus the situation in the cow is similar to that in the sheep and the increased turnover of pregnenolone into estrone may explain the decrease of placental progesterone production in the cow towards the end of gestation.

Analysis of the hyperpolarizing effects of forskolin in guinea-pig atrial heart muscle.

The effects of forskolin on action potential configuration and on both uptake and efflux of 86Rb+ were studied in guinea-pig left atria. The action potential was prolonged by forskolin in the plateau range but shortened at the end of repolarization; maximal upstroke velocity and amplitude of slow response potentials were enhanced. In partially depolarized preparations, the resting potential was increased by forskolin; this effect was not prevented by atropine 1 mumol/l. Forskolin augmented the rate constant of 86Rb+ efflux in beating and in resting preparations. The uptake of 86Rb+ was enhanced by forskolin in resting preparations. It is concluded that forskolin stimulates the Na+,K+-pump and activates a background potassium conductance. Both effects may account for the shortening effect of the drug on the action potential and the increase in resting potential seen in partially depolarized preparations.

[Ambulatory therapy of trichomoniasis vaginalis. Clinical trial of a new Trichomonas-active azide substance]. / Ambulante Therapie der Trichomoniasis vaginalis. Erfahrungsbericht über Ergebnisse der klinischen Prüfung einer neuen trichomonazid-wirksamen Substanz.

Report on the results of the clinical trial of a new trichomonacide in ambulatory patients in a gynaecological practice. A report is given on a new drug for the treatment of vaginal trichomoniasis, which has been tried in 87 patients of a gynaecological practice under outpatient conditions. The rate of failures was 5.7% (5 of 87 cases). The advantages over alternative preparations is to be seen in the short duration of treatment (24 hours at the most), and in the favourable tolerance in spite of high doses.