Femtosecond single-electron diffraction.

Ultrafast electron diffraction allows the tracking of atomic motion in real time, but space charge effects within dense electron packets are a problem for temporal resolution. Here, we report on time-resolved pump-probe diffraction using femtosecond single-electron pulses that are free from intra-pulse Coulomb interactions over the entire trajectory from the source to the detector. Sufficient average electron current is achieved at repetition rates of hundreds of kHz. Thermal load on the sample is avoided by minimizing the pump-probe area and by maximizing heat diffusion. Time-resolved diffraction from fibrous graphite polycrystals reveals coherent acoustic phonons in a nanometer-thick grain ensemble with a signal-to-noise level comparable to conventional multi-electron experiments. These results demonstrate the feasibility of pump-probe diffraction in the single-electron regime, where simulations indicate compressibility of the pulses down to few-femtosecond and attosecond duration.

Surgical therapy of Peyronie's disease by partial plaque excision and grafting with collagen fleece: feasibility study of a new technique.

Surgery is gold-standard for correction of Peyronie's curvature. Grafting is preferred in advanced deviations. We present our novel surgical technique and early results of grafting with collagen fleece. Patients with stable Peyronie's disease (PD) were included. Grafting was performed by a ready-to-use collagen fleece coated with tissue sealant (TachoSil, Nycomed, Konstanz, Germany), following partial plaque excision/incision. Results of correction were documented by artificial erection. In all, n=70 consecutive patients underwent surgery. Mean patient age was 56.4 years (range: 33-72); 88.6% of patients had dorsal deviation, 11.4% lateral or ventral deviation. Grafting after partial plaque excision was performed in 61 patients (87.1%), after plaque incision in 2 (2.9%) patients. In the former patients, mean operative time was 94.2 min (range: 65-165). Totally straightness was achieved in 83.6%. Three patients required surgical drainage because of subcutaneous haematoma formation. After mean early follow-up of 5.2 days (range: 2-15), glans sensation was normal in 56 patients (91.8%). Seven patients (10.0%) underwent Nesbit procedure alone. Grafting by collagen fleece in PD is feasible and promising. Major advantages are decreased operative times and easy application. Moreover, an additional haemostatic effect is provided. However, long-term clinical outcomes are necessary to confirm these encouraging findings.

[Minimally invasive PCNL (mini-perc). Alternative treatment modality or replacement of conventional PCNL?]. / Minimal-invasive PCNL (Mini-Perc). Alternative oder Ersatz für konventionelle PCNL?

Extracorporeal shock wave lithotripsy (SWL) is the current treatment modality of choice for upper urinary tract calculi. For stones with a diameter >2 cm and in lower calyceal stones and diverticular stones, the stone-free rate of SWL is rather poor. In these cases, conventional percutaneous nephrolithotomy (PCNL) leads to an increased stone-free rate, but morbidity with conventional PCNL is significant higher than in SWL. With the invention of miniaturized nephroscopes (mini-perc), new treatment options are available. The experience based on hundreds of treatments using mini-perc show comparable results of mini-perc to conventional PCNL and a complication rate comparable to that for SWL. These favorable results are independent of stone size, stone location, and patient age. In summary, mini-perc can be recommended as a primary approach to stones of the upper urinary tract exceeding 1 cm, larger lower-pole stones, and calyceal diverticular stones. Mini-perc can be regarded as an alternative treatment modality to conventional PCNL.

Diagnosis and metaphylaxis of stone disease. Consensus concept of the National Working Committee on Stone Disease for the upcoming German Urolithiasis Guideline.

This review draws the recent state of the art in metabolic diagnosis and metaphylaxis of stone disease. It is the basis for the consensus approval with the other medical societies and institutions in Germany involved in the guideline process of the new "Urolithiasis Guideline". The German Working Committee on Stone Disease reviewed critically the current literature in the field of urolithiasis-including the existing German and EAU-Guidelines as well as the Conference Book of the First International Consultation on Stone Disease. As far as possible the references were rated according to the EBM criteria. On this basis the expert group discussed all pathways and statements regarding the management of stone disease. The present review coincides with the consented guideline draft of the German Working Committee on Stone Disease. Occurrence of stone disease in the western world increases seriously. Modern lifestyle, dietary habits and overweight-problems of the affluent societies-emerge to be the important promoters of the "stone-boom" in the new millennium. This even affects children, whose stone prevalence is otherwise significantly less than that of adults. Criteria for the high risk group of stone formers were clearly defined. A diagnostic standard is formulated for the basic and the elaborate metabolic evaluation of a stone patient. Approximately 75% of all stone patients could anticipate stone recurrence with elementary reorientation of their lifestyle and dietary habits, summarized as general metaphylaxis. About 25% of the stone formers require additional pharmacological intervention to normalize their individual biochemical risk, precisely compiled for each stone type as specific metaphylaxis.

Renal tubular alteration by crystalluria in stone disease-an experimental study by means of MDCK cells.

OBJECTIVES: Physicochemical properties of urine do not explain the formation of urinary stones. Clinical findings and results of animal experiments suggest that alteration to the renal tubular cell plays a key role in the initiation of urinary stone formation. It is not clear whether this is a primarily intracellular alteration of metabolic origin which, after lysis of the renal tubular cell in the lumen, presents a nucleus for the formation of concretions, or whether in the lumen it is tubular cell damage induced by crystalluria that triggers the formation of urinary stones. MATERIALS AND METHOD: Using Madin-Darby canine kidney cells, the influence of crystalluria on the renal tubular cell was tested in cell cultures. The influence of parathyroid hormone, vitamin D(3), oxalate and calcium concentrations and the extent to which these processes can be inhibited by allopurinol and selenium were investigated. RESULTS: Calcium oxalate monohydrate crystals produced reproducible damage to the renal tubular cell which was independent of parathyroid hormone and vitamin D(3). The crystalluria-induced effects were unrelated to the oxalate and calcium concentration or the pH. Allopurinol and selenium were able to inhibit the processes. CONCLUSION: The results indicate secondary involvement of the renal tubular cell in lithogenesis as a result of luminal alteration caused by calcium oxalate crystals. Mechanical damage and interaction between crystal and tubular cell lead to the apposition of crystals. The nephroprotective effect of allopurinol and selenium as antioxidants might explain the benefit of allopurinol found clinically in terms of stone metaphylaxis.

Genetic variations of the SLC7A9 gene: allele distribution of 13 polymorphic sites in German cystinuria patients and controls.

Cystinuria is a hereditary disorder of cystine and dibasic amino acid transport across the luminal membrane of renal tubules and intestine, resulting in recurrent nephrolithiasis. While mutations in the SLC3A1 gene cause type I cystinuria, patients with non-type I cystinuria carry mutations in the SLC7A9 gene. Both gene products form the renal amino acid transporter rBAT/b0,+AT affected in cystinuria. In the present study a total of 59 patients with different ethnic background were screened for sequence variations in SLC7A9, out of these 32 were of German origin. For determination of allele frequencies of detected polymorphisms, 58 healthy German controls were investigated. Molecular-genetic analysis was performed using single-strand conformation polymorphism analysis, restriction assays and sequencing. Allele frequencies were analyzed statistically for the detected polymorphisms. In addition to the 6 already known variants we identified 7 new polymorphisms. Statistical analyses showed a significantly different distribution of alleles between German patients and German controls in case of the polymorphisms c. 147C>T (exon 2), c.386C>T (exon 3), IVS3+22T>G, c.584C>T (exon 4), c.610T>C (exon 4), c.692C>T (exon 5), c.852C>A (exon 6) and c.872C>T (exon 6). In summary, our results show that cystinuria is a complex disease which is not only caused by mutations in SLC7A9 and SLC3A1, but also influenced by other modifying factors such as variants in SLC7A9.

Urinary infection stones.

Infection stones make up approximately 15% of urinary stone diseases and are thus an important group. These stones are composed of struvite and/or carbonate apatite. The basic precondition for the formation of infection stones is a urease positive urinary tract infection. Urease is necessary to split urea to ammonia and CO(2). As a result, ammonia ions can form and at the same time alkaline urine develops, both being preconditions for the formation of struvite and carbonate apatite crystals. When these crystals deposit themselves infection stones form. If these infections are not treated and the stones are not removed, the kidney will be damaged. For stone removal modern methods are available, e.g. ESWL and/or instrumental urinary stone removal. Here especially less invasive methods are preferable. Any treatment must be adjusted to the patient individually. Patients should be examined frequently for recurrent urinary tract infections and stone recurrences and, newly arising infections must be resolutely treated. Good therapy and prophylaxis are possible with present-day treatment modalities.

Collagen fleece for defect coverage following plaque excision in patients with Peyronie's disease.

OBJECTIVES: Surgical correction of penile deviation in patients with Peyronie's disease by tunical plication often leads to shortening of the penis. It is, thus, recommendable to combine tunical plication with plaque incision or excision. The resulting tunical defect, however, requires grafting, and various techniques have been described. In comparison with tunical plication, all of these combined techniques are associated with increased operation time, mainly due to additional procedures necessary at the donor site, and may result in defects at the donor site. We here report a novel surgical technique by which tunical defects after partial excision of plaques are covered by a ready-to-use collagen fleece coated with tissue sealant (TachoComb). METHOD: A prospective clinical observation trial was conducted in 19 patients with penile deviation due to Peyronie's disease. RESULTS: In all patients, a reliable closure of the Tunica albuginea was achieved, and no postoperative haematoma formation was observed. Postoperatively, none of the patients suffered from erectile dysfunction. During the follow-up period of 25 months, objective and subjective improvement was 83% and 72%, respectively. CONCLUSION: The present data indicate that this novel surgical technique may be of benefit in patients with Peyronie's disease and should thus be further evaluated.

Analysis of the genes SLC7A9 and SLC3A1 in unclassified cystinurics: mutation detection rates and association between variants in SLC7A9 and the disease.

Cystinuria is a common inherited disorder of defective renal reabsorption of cystine and dibasic amino acids. Recently, 2 responsible genes have been identified: mutations in the SLC3AI gene encoding the glycoprotein rBAT cause cystinuria type I, while variants in the SLC7A9 gene have been demonstrated in non-type I cystinuria; its gene product b(0)+AT is the light chain of the renal cystine transport system rBAT/b(0),+-AT. To estimate the role of both genes in the etiology of cystinuria, we searched for sequence alterations in SLC7A9 and SLC3AI: 30 unclassified cystinurics were investigated. In 50% of patients (15/30), point mutations in SLC3A1 were detected. Screening of the SLC7A9 gene revealed 10 mutations in 8 patients corresponding to a frequency of 27%. In addition to previously published mutations in the SLC7A9 gene, we detected 2 new mutations (F 140S, c747delG). An overall detection rate of 73% (22/30) in unclassified patients is delineated for mutations in both genes. In 33% (10/30), 2 mutations were detected, in 40% (12/30) 1 mutation. Furthermore, 5 new polymorphic sites were identified in SLC7A9. While the base pair variation in intron 9 is homogeneously distributed in patients and control individuals, the allelic and genotypic distributions of the polymorphisms in 3 exons of SLC7A9--exons 2, 5 and 6--and intron 3 differ significantly between both groups. Our results suggest that some haplotypes defined through the exons 2, 5 and 6 and intron 3 might be markers of a functional variant in the SLC7A9 gene. Evidently, since the mutation detection rates in the 2 so far known cystinuria genes never reach 100%, further genes and modulating factors should influence the phenotype in a subset of patients. However, the presented data show that testing for mutations in the 2 currently known cystinuria genes is already a meaningful approach to the molecular diagnostics of the disease.

Comparison of cytology and nuclear matrix protein 22 for the detection and follow-up of bladder cancer.

OBJECTIVES: This study was designed to determine the clinical usefulness of the nuclear matrix METHODS: 84 patients suffering from bladder cancer or suspected bladder cancer, 25 patients with benign urological lesions and 60 healthy controls participated in a prospective study. Freshly voided spot urine samples were taken for cytological examination and determination of NMP 22 levels by enzyme-linked immunoassay. RESULTS: The sensitivity of the NMP 22 test according to the tumor grading was (results of cytology in brackets): G1 25.0% (20.0%); G2 68.2% (59.1%), and G3 100.0% (66.7); overall sensitivity was 62.5% (45.0%). The sensitivity for superficial bladder cancer was 46.7% (36.7%) and for invasive bladder cancer 90.0% (70.0%). The specificity was 65.9% (88.9%). CONCLUSIONS: NMP 22 is a reliable tool for detecting invasive bladder cancer. Results for the frequently occurring low grade superficial bladder cancer are as poor as those obtained with cytology. In addition benign lesions such as urolithiasis or urinary tract infection lead to false-positive results. Therefore cystoscopy has to be performed when trying to detect and follow-up bladder cancer.

[Molecular biology of cystinuria]. / Molekularbiologie der Zystinurie.

Cystine stones belong to the kidney stones diseases which are difficult to manage due to the high rate of recidivation and the necessary surgical procedures. Cystinuria is an autosomal recessive defect of the transport of cystine and the dibasic amino acids in the proximal kidney tube. Recently, two subunits of a renal cystine transporter have been identified. Mutations in these genes have been shown to lead to the cystinuria phenotype. Genetic and functional analyses have helped to modify the classification of cystinuria which had previously been exclusively based on biochemical data. Furthermore, first steps towards a molecular genetic testing have been carried out replacing the so far diagnostic procedure which are stressing for the patients. The results of these testings make an individual therapy possible.

Minimally invasive PCNL in patients with renal pelvic and calyceal stones.

Stones of the renal pelvis can be treated either by extracorporeal shock wave lithotripsy (SWL) or percutaneous nephrolithotomy (PCNL). As a low-risk procedure with a longer treatment period, SWL often leads to persistent residual stone fragments, whereas conventional PCNL achieves a higher stone-free rate and allows a shorter treatment period albeit with a somewhat higher surgical risk. To reduce the invasiveness of conventional PCNL, the application of a miniaturised instrument for PCNL (MPCNL) was evaluated. For MPCNL a rigid nephroscope with a calibre of 12 F was developed and used in 19 patients. After puncture of the kidney under ultrasound control and single-step dilatation, a 15 F Amplatz sheath was placed. Data on the stone size and location, stone-free rate, blood transfusions, operating time and complications were recorded. In all patients, the part of the kidney afflicted by the stone was successfully punctured. On average, retreatment rate was 0.7. The mean stone size was 2.4 cm(2). The average operating time was 99.2 min. In every case, the absence of residual stones was confirmed radiologically and nephroscopically. Hemorrhages requiring a blood transfusion did not occur. A febrile pyelonephritis occurred as a postoperative complication in one patient (= 5.3%). MPCNL represents an alternative to SWL for renal calculi with a size from 1 to 2 cm located in the renal pelvis and calices, especially the lower calix. The advantages are the short treatment time, the high stone-free rate and the accessibility of lower pole stones which are less amenable to SWL. MPCNL is not suitable for large concrements since the limited sheath diameter would increase the operating time. Due to this limitation, MPCNL represents an extension of the indication for conventional PCNL that it can in no way replace.

Arterial blood pressure following different types of urinary stone therapy. Presented at the 8th European Symposium on Urolithiasis, Parma, Italy, 1999.

OBJECTIVE: Several studies reported increased blood pressure (BP) values following extracorporeal shock wave lithotripsy (ESWL) treatment of renal stones. It is unclear, however, whether this is due to ESWL, since nephrolithiasis itself increases the relative risk of developing hypertension. Therefore we prospectively studied the BPs of stone patients undergoing different types of treatment. METHODS: 252 stone patients (63% males, 37% females, median age 44.3, range 11.7-86.4 years) participated. 168 suffered from uretral stones: 50 underwent ESWL; 40 ureteroscopy, and 78 patients passed stones spontaneously (SP). 84 had renal stones: 60 underwent ESWL; 8 percutaneous nephrolithotomy/open surgery, and 16 no treatment. Systolic (SBP) and diastolic (DBP) BP were measured according to Riva-Rocci prior to, immediately after, and 3, 6, 12, 18 and 24 months after stone therapy. RESULTS: Immediately after SP, SBP decreases, whereas after active stone treatment increases (highest after ESWL) in SBP were seen. DBP was unchanged. During the further follow-up, a gradual increase in BP was observed in all groups. At 24 months in all groups, regardless of the stone location and type of treatment, SBP and DBP were significantly higher than the pretreatment levels (p = 0.000). There was no a difference between renal and ureteral stones, or between the ESWL treatment and the other groups. CONCLUSION: Renal stone disease itself rather than the type of treatment significantly increases SBP and DBP during a follow-up period of 24 months. The underlying mechanisms remain to be elucidated.

Reduction of high-energy shock-wave-induced renal tubular injury by selenium.

In shock-wave-induced renal injury cavitation-generated free radicals play an important role. Using an in vitro model with Madin-Darby canine kidney (MDCK) cells, we investigated the influence of selenium, a free radical scavenger, in shock-wave-induced tubular cell injury. Suspensions of MDCK cells (33 x 10(6) cells/ml) were placed in small containers (volume 1.1 ml) for shock wave exposure. Two groups of 12 containers each were examined: (1) control (no medication), (2) selenium (0.4 microg/ml nutrient medium). Six containers in each group were exposed to shock waves (impulse rate 256, frequency 60 Hz, generator voltage 18 kV), while the other six containers in each group served as a control. After shock wave exposure, the concentration of cellular enzymes such as lactate dehydrogenase (LDH), N-acetyl-beta-glucosaminidase (NAG), glutamate oxaloacetate transaminase (GOT) and glutamate lactate dehydrogenase (GLDH) in the nutrient medium was examined. Following shock wave exposure there was a significant rise in LDH, NAG, GOT and GLDH concentrations. Selenium reduced this enzyme leakage significantly. Thus we conclude that selenium protects renal tubular cells against shock-wave-induced injury. Since selenium is an essential part of glutathione peroxidase, this effect seems to be mediated by a reduction in reactive oxygen species.

Control of hepatic parameters in renal cell carcinoma (RCC) by interleukin-6 (IL-6)?

RCC can go hand-in-hand with an elevation of various hepatic proteins. An interrelationship between the IL-6 titer, C-reactive protein (CRP) and the blood sedimentation rate (BSR) has already been proven. The aim of the present study was to study 1) the possibility of differentiating between healthy and RCC patients via IL-6 in the serum and 2) the relationship of IL-6 to hepatic parameters {alkaline phosphatase (AP), gamma-glutamyltransferase (gGT), serum proteins (E'p)} and the usual clinical prognostic parameters (tumor grading, staging). Serum analysis of 38 healthy patients via ELISA (DPC-Biermann, Germany) showed normal values of 1.2 ng/ml for IL-6, with a standards deviation of +/- 1.7 and a peak concentration of 3 ng/ml (specificity: 95%). In 20 RCCs there were IL-6 titers of 10.7 ng/ml +/- 6.56 in the pre-operative serum. The sensitivity of IL-6 was about 90%. The difference was statistically significant (p < 0.0001, Wilcoxon test). For IL-6 there was a positive correlation with the BSR (1-hour value: r = 0.7; 2-hour value: r = 0.6), CRP (r = 0.85), E'p (r = 0.6), and gGT (r = 0.6). No correlation was found between AP, the Robson stage, grading, and IL-6. IL-6 is potentially suitable for differentiating between healthy and RCC patients but is not tumor specific. IL-6 has a strong correlation with all laboratory values which were analyzed except AP thus there is considerable evidence for a cytokine (IL-6) control of the hepatic changes. Since some of the above-named laboratory parameters have prognostic relevance, IL-6 can be regarded as a cumulative prognostic parameter.

Mutations in the SLC3A1 gene in cystinuric patients: frequencies and identification of a novel mutation.

Cystinuria is a frequent autosomal recessive transport disorder characterized by defective renal resorption of cystine and other dibasic amino acids. Biochemically, three types of cystinuria can be defined. Here we present our results of screening for mutations in the SLC3A1 gene, which codes for a dibasic amino acid transporter protein and appears to be involved in the pathogenesis of cystinuria type I. Our study population consists of 5 Italian cystinuria type I patients and 10 cystinuric patients as yet unclassified as to clinical type. The latter were of different ethnic origin. In total, we found 13 point mutations and 8 genomic rearrangements in 15 cystinuric patients, i.e., our detection rate was 70% (23/30 chromosomes). Remarkably, in patients known to be suffering from cystinuria type I, the mutation detection rate was only 50%, whereas in patients unselected as to cystinuria type, we found 80% of mutations. Additionally, our results, as with those published in the literature, indicate a possible population specific distribution of mutations: Each of the 4 Greek patients analyzed here showed homozygosity for mutation T216M in exon 3. Analysis of a Yugoslavian patient showed homozygosity for a novel mutation, R365L, in exon 6 (nt1094G > T). Findings from molecular genetic studies, as well as physiological investigations, suggest that there are further genes that play a role in the etiology of cystinuria. Nevertheless, our results show that screening for mutations in the SLC3A1 gene can be a meaningful step toward molecular genetic diagnosis of cystinuria in patients without biochemical classification. As with cystic fibrosis, the finding of specific mutations in particular ethnic populations, suggest that the diagnostic approach should take into consideration a patient's ethnic origins.

Renal cell carcinoma: relevance of angiogenetic factors.

Angiogenesis is essential for tumour growth. Mostly hypervasculariced renal cell carcinoma (RCC) usually express angiogenic factors, e.g. basic fibroblast factor (bFGF), vascular endothelial growth factor, and angiogenin (ag). This study was designed to evaluate the improvement of serological angiogenesis-factors, their prognostic relevance and correlation to tumour-grade, -stage and -volume. Measurement of bFGF, vEGF and ag was done with ELISA (Quantikine), R and D-Systems Europe Abbington, United Kingdom. The control group comprised 39 healthy blood donors, RCC-group were 35 patients with different RCC. Survival dates were presented with Kaplan-Meyer-curves, significance was tested with students-t-test. For all angiogenic factors a highly significant difference between the healthy and RCC-group was found. A strong correlation between angiogenic factors and tumor grade, -stage and -volume, was not found, but a trend for each of the angiogenic factors to correlate with grade was seen. No survival benefit was seen between patients with normal angiogenic factor over those with elevated angiogenic levels.

[Collagen metabolism in induratio penis plastica (IPP)]. / Untersuchungen zum Kollagenstoffwechsel bei Induratio penis plastica (IPP).

The exact pathogenesis of Peyronie's Disease (PD) is still unknown. Histopathological investigations suggest, that the metabolism of the collogenious tissue plays an important role. Therefore, we started a prospective study in order to evaluate the metabolic situation of 11 patients with PD and 11 healthy controls. We measured biochemical parameters of penile and venous blood, which correspond to the collagen metabolism (procollagen III [P III P], PMN-elastase [PMN-E], fibronectin [FN], fibronectinreceptor [FN-R] und alpha-1-proteinase-inhibitor [alpha-1-PI]). There was no difference in PMN-E, P III P, FN and FN-R in all samples. There was no difference in penile and venous blood as well. But we found out a highly significant lower concentration of alpha-1-PI in patients with PD (158,55 mg/dl) than in controls (214,82 mg/ dl). A deficiency of alpha-1-PI can lead to a higher activity of serum proteases, such as PMN-E. This can result in a change in collagen metabolism by an increased synthesis of collagen type III, that is found in PD. This general pathophysiologic factor may be competed by an additional local alteration of the tissue.