S.A. Loening

Cathepsins B, H, L and cysteine protease inhibitors in malignant prostate cell lines, primary cultured prostatic cells and prostatic tissue

Elevated activities of cysteine proteinases, the cathepsins B, H, L (CB, CH, CL) and diminished cysteine protease inhibitors (CPI) have been demonstrated in a variety of tumours and have been suggested to contribute to invasion and metastasis. The situation for prostate cancer is still unknown. In this study, using fluorimetric assays, the catalytic activities of CB, CH, CL were measured in prostatic tissue samples after radical prostatectomy, adenomectomy, transurethral resection of the prostate, in cell cultures grown from cancerous and non-cancerous parts of human prostate after prostatectomy and in the cell lines LNCaP, DU 145 and PC 3. CPIs were determined using heat activation before testing their inhibitory activity against purified CB. Comparing matched pairs of normal and cancerous tissue samples from the prostate, significantly decreased levels of CB, CL in malignant parts of the prostate were found. In contrast, primary cell cultures from cancerous samples showed elevated levels of CB, CH, CL and increased ratios of cathepsins to CPI compared with cell cultures from normal prostate. Established cell lines showed a similar distribution pattern of each cathepsin, DU 145 containing the highest levels, followed by LNCaP and PC 3. Our results suggest that elevated cathepsin levels and consequently increased ratios of cathepsins to CPI in primary cell cultures from cancerous versus non-cancerous parts of the prostate may be indicative of a cellular proteolytic imbalance in prostatic cancer cells. In this respect, primary cell culture experiments should be preferred to determinations in tissue samples.

RATIO OF FREE-TO-TOTAL PROSTATE SPECIFIC ANTIGEN IN SERUM CANNOT DISTINGUISH PATIENTS WITH PROSTATE CANCER FROM THOSE WITH CHRONIC INFLAMMATION OF THE PROSTATE

PURPOSE We demonstrate the effect of chronic inflammation of the prostate on the ratio of free-to-total prostate specific antigen (PSA) in serum calculated as a percentage of free PSA and, therefore, that percentage of free PSA is an unspecific means to distinguish among prostate cancer, chronic prostatitis and benign prostatic hyperplasia (BPH). MATERIALS AND METHODS Total, free and percentage of free PSA was measured in 66 men with prostate cancer, 119 with BPH and 17 with asymptomatic chronic prostatitis. In all patients the diagnosis was histopathologically confirmed by microscopic examination of prostatic specimens after sextant biopsy, transurethral prostatic resection or prostatectomy. RESULTS The median values of total, free and percentage of free PSA were 4.11 microg./l., 0.75 microg./l. and 20.4% in patients with BPH, 10.0 microg./l., 0.84 microg./l. and 8.5% in those with prostate cancer, and 7.60 microg./l., 1.23 microg./l. and 10.6% in those with chronic prostatitis. Patients with prostate cancer and chronic prostatitis had a significantly lower percentage of free PSA than those with BPH. Receiver operating characteristics curve analysis showed that percentage of free PSA as a discriminator between prostate cancer and BPH was not suitable for differentiating between prostate cancer and chronic prostatitis. CONCLUSIONS Chronic prostatitis is not characterized by elevated total PSA concentrations alone but also by a decreased percentage of free PSA, a tendency similar to that in prostate cancer. This unspecific change in percentage of free PSA must be considered to interpret the percentage of free PSA correctly.

Factors influencing the ratio of free to total prostate-specific antigen in serum

The ratio of free prostate‐specific antigen (f‐PSA) to total PSA (t‐PSA) in serum, calculated as percent free PSA (f‐PSA%), is lower in patients with prostate carcinoma (PCa) than in patients with benign prostate hyperplasia (BPH). This parameter facilitates discrimination between the 2 groups of patients, but there is an overlapping of data. A better understanding of factors influencing this ratio is of practical importance. Therefore, f‐PSA% was measured in controls and patients suffering from BPH, PCa and chronic prostatic inflammation with t‐PSA concentrations up to 20 μg/l using the IMMULITE assays. The relationships of f‐PSA% to clinical situation, age, prostate volume, kind of treatment, and stage and grade of tumor were calculated. Compared with controls or BPH patients, mean f‐PSA% values were reduced in PCa patients and in patients with chronic prostatic inflammation. The prostate volume was the most important factor to influence f‐PSA%. The difference of f‐PSA% between PCa and BPH patients with prostate volumes smaller than 40 cm3 was lost if the prostate volumes exceeded 40 cm3. No relationship of f‐PSA% to pTNM stage or grade of tumor was observed. In contrast to t‐PSA concentrations, the f‐PSA% values were not age‐dependent and were not influenced by any kind of treatment in BPH and PCa patients either, which simplifies the use of f‐PSA% compared with t‐PSA. Thus, for using f‐PSA% in clinical practice and for interpreting the data correctly, the advantages shown have to be considered along with the potential limitations of f‐PSA%. Int. J. Cancer 74:630–636.© 1997 Wiley‐Liss, Inc.

Die laparoskopische radikale Prostatektomie Erfahrungen mit 145 Eingriffen

ZusammenfassungErmutigt durch die ersten erfolgreichen Berichte über die laparoskopische radikale Prostatektomie haben wir diesen Eingriff im Juni 1999 in unser Operationsprogramm aufgenommen. Bis Ende November 2000 führten wir insgesamt 145 dieser Eingriffe durch. Die Indikation zur laparoskopischen Prostatektomie entsprach genau der, die wir für die offene Operation anwenden.Abdominelle Voroperationen, transurethrale Resektionen bzw. eine mäßige Adipositas sind keine Kontraindikationen. Eine Konversion zur offenen Operation war in keinem Fall erforderlich. Die postoperative Komplikationsrate betrug 11,7%, wobei es sich vorwiegend um Minorkomplikationen handelte. Auch hinsichtlich der Kontinenz und Potenz sind die Ergebnisse sehr gut.Unserer Meinung nach ist die laparoskopische radikale Prostatektomie eine echte Alternative zur offenen Operation, da sie eine Reihe von Vorteilen für den Patienten und den Operateur aufweist.AbstractEncouraged by the groups in Paris, we performed 145 laparoscopic radical prostatectomies between June 1999and the end of November 2000. The indication for laparoscopic prostatectomy is the same as for open surgery: an organ-confined cancer.Previous abdominal surgery, transurethral resection, and/or relative adiposity are not considered to be contraindications for this laparoscopic procedure. The mean operating time was 255 min; the last 60 procedures took an average of 200 min. In no case was it necessary to convert to open surgery. Worthy of note was the low blood loss of 185 ml on average so that in 98% of the patients no blood transfusion was required. After completing the learning curve, the average indwelling catheter time was only 5.5 days. The postoperative complication rate was 11.7%, consisting mainly of minor complications. Also with regard to continence and potency, the results were representative. Postoperatively, 75%, 86%, 92%, and 93% of the patients were continent after 3, 6, 9, and 12 months, respectively.In our opinion, laparoscopic radical prostatectomy is an alternative to open prostatectomy, offering a number of advantages for the patient and surgeon as well.

Laparoscopic Lymphocele Drainage After Renal Transplantation

Lymphoceles are among the most frequent surgical complications after renal transplantation. We performed laparoscopic drainage in 5 patients with a large lymphocele in the small pelvis after successful kidney transplantation.

Laparoskopische radikale zystektomie mit intrakorporaler anlage einer kontinenten harnableitung. Zukunft oder gegenwart

ZusammenfassungSind ausreichend Erfahrungen mit der laparoskopischen Prostatektomie vorhanden, ist der Schritt zur laparoskopischen Zystektomie nicht mehr sehr groß. Die eigentliche Herausforderung besteht in der laparoskopischen Konstruktion der Harnableitung.Wir berichten in diesem Artikel über Erfahrungen mit 11 laparoskopischen Zystektomien und der intrakorporalen Anlage eines Mainz Pouch II als kontinente Harnableitung bei Patienten mit einem muskelinvasiven Harnblasenkarzinom. Alle 11 Eingriffe konnten erfolgreich durchgeführt werden. Eine Konversion war nicht notwendig. Die mittlere Operationszeit betrug 6,7 h. Bis auf 2 Pouchfisteln wurden keine Komplikationen beobachtet. Sowohl die funktionellen als auch onkologischen Ergebnisse sind überzeugend. Der wesentliche Vorteil des minimal invasiven Vorgehens ist die deutlich reduzierte Morbidität mit kürzerer Rekonvaleszenz. Außerdem senkt der extrem niedrige Blut-, Elektrolyt- bzw. Flüssigkeitsverlust den kardiovaskulären Stress enorm.Die laparoskopische radikale Zystektomie stellt derzeit die Grenze der technisch machbaren und sinnvollen Laparoskopie dar und sollte deshalb den Zentren mit entsprechender Expertise vorbehalten bleiben.AbstractOnce laparoscopic radical prostatectomy has been mastered, the step to performing a radical cystectomy is not that far. The challenge is to create the urinary diversion by laparoscopy.In this report we describe our experience with 11 laparoscopic radical cystectomies and intracorporeal construction of a continent urinary diversion (Mainz pouch II) as a treatment option in patients with muscle-invading bladder cancer. All 11 procedures could be performed successfully. A conversion to open surgery was not required in any case. The mean surgery time was 6.7 h. Except for two pouch fistulas we did not observe any intra- or postoperative complications. The functional as well as the oncological results are convincing. Less morbidity and faster recovery are the main advantages of this minimally invasive procedure. In addition, the low levels of blood loss, fluid shifts, and electrolyte loss considerably reduce cardiovascular stress.Radical cystectomy and construction of a continent urinary diversion represent the limit of technically feasible laparoscopy and should be done exclusively in specialized centers.

Nonmhyperbolic calcium calibration curve of Fura-2: implications for the reliability of quantitative Ca2+ measurements

The fluorescence probe Fura-2 is widely applied for the quantitative determination of cellular free Ca2+ concentration. Generally, a hyperbolic calibration curve has been taken as a basis. The in vitro calibration of Fura-2 performed with free Ca2+ concentrations ranging from 10 nM to about 9 mM demonstrates a non-hyperbolic curve. Assuming two Ca2+ binding sites of Fura-2 dissociation constants of 190 nM and 176 microM were estimated. The analysis of the calibration data by the equation introduced by Grynkiewicz et al. [Grynkiewicz G., Poenie M., Tsien R.Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 1985; 260: 3440-3450] shows that the inclusion of Ca2+ concentrations in the range of the low-affinity binding site leads to an overestimation of the dissociation constant normally used within this method. The Rmax value conventionally estimated at a nominal saturating free Ca2+ concentration increases if concentrations in the range of the low-affinity binding site are selected for this purpose. Provided that millimolar free Ca2+ concentrations are applied for the determination of Rmax, the experimentally-estimated free Ca2+ concentration should be considerably lower than the real cellular concentration. This bias is especially to be expected if free Ca2+ concentrations exceeding the high-affinity Ca2+ binding site of Fura-2 are measured. With the use of approximately 10 microM free Ca2+ for the determination of Rmax the contribution of the low-affinity binding site is negligible. Data for cellular free Ca2+ calculated on the basis of Rmax values estimated with Ca2+ concentrations in the millimolar range should be considered with caution.

Thermoradiotherapie mit interstitiellen Thermoseeds bei der Behandlung des lokalen Prostatakarzinoms Erste Ergebnisse einer Phase-II-Studie

ZusammenfassungIm Rahmen des Sonderforschungsbereichs Hyperthermie (273) der Deutschen Forschungsgemeinschaft wurde eine Phase-II-Studie initiiert, um die Effektivität der interstitiellen Hyperthermie in Kombination mit konformaler Strahlentherapie beim lokalen Prostatakarzinom zu ermitteln. Zwischen Juli 1997 und April 2000 konnten 41 Patienten rekrutiert werden.Die interstitielle Hyperthermie wurde mittels Kobalt-Palladium-Seeds in einem magnetischen Feld erzeugt. Die perkutane konformale Strahlentherapie begann simultan zur Hyperthermie. Die tägliche fraktionierte Dosis betrug 1,8 Gy und die Gesamtdosis 68,4 Gy. Während der Therapie kam es zu keiner Grad-III- oder -IV-Nebenwirkung. Der mediane initiale PSA-Wert fiel in einem medianen Beobachtungszeitraum von 10 Monaten von 11,25 ng/ml auf 0,88 ng/ml 3 Monate nach Therapie und auf 0,38 ng/ml 12 Monate nach Therapie ab.Wir konnten nachweisen, dass die interstitielle Hyperthermie mit Thermoseeds durchführbar und die Kombination mit der konformalen Strahlentherapie verträglich ist. Ein längerer Beobachtungszeitraum ist erforderlich, um die Effizienz dieser Kombination zu beurteilen.AbstractSeveral experimental data are available regarding the efficacy of the combination of interstitial hyperthermia and conformal radiation therapy. We planned a phase II trial as a member of the special hyperthermia research group of the Deutsche Forschungsgemeinschaft (German Research Society) to determine the efficacy of thermoradiotherapy using interstitial cobalt-palladium thermoseeds in the treatment of patients with localized prostate cancer.Forty-one patients with localized prostate cancer were enrolled in the study between July 1997 and April 2000. The interstitial hyperthermia induced in a magnetic field was applied in six sessions once a week. Conformal three-dimensional radiation therapy was given simultaneously in daily fractions of 1.8 Gy with a total dose of 68.4 Gy. We measured intraprostatic temperatures between 42° and 46°C. No major side effects were observed during the hyperthermia session. The median level of prostate-specific antigen (PSA) decreased from 11.25 ng/ml to 0.88 ng/ml 3 months after treatment and to 0.38 ng/ml 12 months after treatment with a median follow-up of 10 months. The mean prostate volume decreased from 32.6 ml to 26 ml after 3 months of treatment and to 18.5 ml after 12 months.Interstitial hyperthermia is a feasible, well-tolerated procedure in prostate cancer therapy. A significant PSA decrease was observed. Longer follow-up is necessary to determine the efficacy.

Brachytherapie des lokalen Prostatakarzinoms

ZusammenfassungDie Brachytherapie des Prostatakarzinoms hat in den letzten Jahren auch in Deutschland mehr Anwendung gefunden. Dabei kommen verschiedene Strahlenquellen in Frage. Diese unterscheiden sich teilweise aufgrund ihrer physikalischen Eigenschaften. Es werden permanente oder temporäre Strahlenquellen angewendet. Permanente Quellen sind 125Jod, 103Palladium und 198Gold. Als temporäre Quelle ist 192Iridium vorhanden. Der Erfolg der Brachytherapie ist von der Patienten- und der Isotopenselektion abhängig. Die LDR-Brachytherapie als Monotherapie eignet sich für Low-risk-Patienten mit initialem PSA-Wert ≤10 ng/ml, Tumorstadium maximal T2b und G2 bzw. Gleason-Score ≤6. Bei Patienten mit einem höherem Tumorstadium muss eine Dosiseskalation stattfinden. Ein synergistischer Effekt einer zusätzlichen perkutanen Strahlentherapie oder einer antiandrogenen Therapie zur LDR-Implantation ist noch nicht bewiesen.AbstractBrachytherapy of prostate cancer has become attractive in recent years in Germany. There are several radioactive sources available, which are physically different. Some of them are used as permanent or temporary implants. The permanent sources most frequently used are iodine 125, palladium 103, and gold 198. Iridium is a temporary implant.The techniques used in Germany are low-dose rate (LDR) and high-dose rate brachytherapy, which differ in dose distribution and patient population.The success of prostate cancer brachytherapy depends on patient selection and choosing the right source for the technique used. Best suited for LDR monotherapy is the low-risk patient with a prostate-specific antigen (PSA) level below 10 ng/ml, maximal tumor stage T2b, and a Gleason score of less than 7.

Molekulare Formen des prostataspezifischen Antigens und des humanen Kallikreins 2 als mögliche Indikatoren in der Prostatakarzinomdiagnostik

ZusammenfassungDas prostataspezifische Antigen (PSA) ist die wichtigste Kenngröße in der Diagnostik und Therapieüberwachung des Prostatakarzinoms. Zur besseren Abgrenzung zwischen benigner Prostatahyperplasie (BPH) und Prostatakarzinom, aber auch zur Früherkennung des Prostatakarzinoms haben sich die molekularen Formen des PSA und neuerdings auch das humane Kallikrein 2 (hK2) als wertvolle Entscheidungshilfen erwiesen.Es wird eine Literaturübersicht über bisher vorliegende Informationen zur Aussagekraft des freien PSA (fPSA) in Relation zum Gesamt-PSA (tPSA), des an α1-Antichymotrypsin gebundenen PSA (ACT-PSA) und des komplexierten PSA (cPSA) zusammen mit eigenen Resultaten gegeben.Der Quotient fPSA/tPSA (fPSA%) hat sich bereits als eine wichtige Entscheidungsgröße in der urologischen Praxis etabliert, mit der Sensitivität und Spezifität der Prostatakarzinomdiagnostik verbessert werden. Die Zahl von Prostatastanzbiopsien im tPSA-Bereich 4–10 μg/l kann damit reduziert und bei tPSA-Werten <4 μg/l können zusätzlich unerkannte Karzinompatienten identifiziert werden. Ein diagnostischer Vorteil der alleinigen Bestimmung des gebundenen PSA bzw. der entsprechenden Quotienten (ACT-PSA/tPSA oder cPSA/tPSA) im Vergleich zum fPSA% konnte bisher nicht sicher nachgewiesen werden. Die Bestimmungen der anderen molekularen PSA-Formen und des hK2 sind noch weitgehend Gegenstand der Forschung bzw. erfordern noch entsprechende klinische Evaluierungen.AbstractProstate-specific antigen (PSA) is the most useful marker in the early detection of prostate cancer and in the monitoring of patients with this diagnosis. Molecular forms of PSA and human kallikrein 2 (hK2) have been used to discriminate between benign prostatic hyperplasia and prostate cancer, as well as for the detection of prostate cancer within the gray zone of PSA.In this respect, a literature survey on the diagnostic validity of free PSA (fPSA) related to total PSA (tPSA), PSA bound to α1-antichymotrypsin (ACT-PSA), and complexed PSA (cPSA) is given together with our own results.The ratio of fPSA/tPSA has been shown to improve both sensitivity and specificity of prostate cancer diagnosis based on tPSA measurements. The number of biopsies can be reduced in the total PSA range of 4–10 μg/l. Furthermore, carcinomas can be detected in patients with PSA values less than 4 μg/l. ACT-PSA or cPSA alone and the calculated derivatives are not superior in their discriminatory power compared with tPSA and the fPSA% value. The other molecular PSA forms and hK2 are still objects of research and their diagnostic significance needs to be evaluated in more extensive clinical trials.