Academic relationships between Hungarian professors and the Second Vienna Medical School.

This article discusses the impact of the 'second' Vienna Medical School, hallmarked by Karl Rokitansky, Joseph Skoda and Ferdinand Hebra, on the study and practice of medicine in Hungary. Six medical doctors' lives and achievements are outlined, who formed a bridge between Vienna and Budapest through their studies and work. Four of them returned to Hungary and promoted the cause of medicine and medical education there. Lajos Arányi (1812-1877) founded in 1844 the Institute of Pathology at the University of Pest. János Balassa (1814-1868) took the Chair of the Surgical Department. Ignaz Philip Semmelweis (1818-1865), the 'Saviour of Mothers', received a position at the Department of Obstetrics and Gynaecology in Vienna in 1846. Gustav Scheuthauer (1832-1894) became Arányi's successor. Each of them continued to keep contact with their tutors in Vienna, especially with Karl Rokitansky, and followed the clinicopathological conception pioneered by the Vienna Medical School regarding diagnostics, treatment and prevention of diseases. Two physicians remained in Vienna: Mór Kaposi (1837-1902), who became known worldwide posthumously due to the connection between Kaposi's sarcoma and AIDS, was the director of the Department of Dermatology of the Vienna University in 1878. Salomon Stricker (1837-1898) undertook the leadership of the Department of General and Experimental Pathology in 1872.

Effect of formaldehyde on cell proliferation and death.

Formaldehyde (HCHO) may reach living organisms as an exogenous agent or produced within cells. The so-called formaldehydogenic compounds like S-adenosyl-L-methionine, N-hydroxymethyl-L-arginine, 1'-methyl ascorbigen, methanol, E-N-trimethyl lysine and methylamine are special exogenous sources of HCHO. Endogenous HCHO can be formed from hydroxymethyl groups during enzymatic methylation and demethylation processes. HCHO, as a highly reactive compound, is considered to be involved in the induction of apoptosis, consequently in the pathogenesis of atherosclerosis and neurodegenerative processes. The biological action of HCHO is dose-dependent. In vitro studies on tumour cell and endothelial cell cultures showed that HCHO in the concentration of 10.0 mM caused necrotic cell death, 1.0 mM resulted in enhanced apoptosis and reduced mitotic activity, while 0.5 and 0.1 mM enhanced cell proliferation and reduced apoptotic activity. Among formaldehydogenic compounds N-hydroxymethyl-L-arginine, 1'-methyl ascorbigen and the HCHO donor resveratrol may be considered as potential inhibitors of cell proliferation. Endogenous HCHO in plants apparently play a role in regulation of apoptosis and cell proliferation. The genotoxic and carcinogentic effects of HCHO is due to production of DNA-protein cross-links. Low doses of HCHO, reducing apoptotic activity may also accumulate cells with such cross-links. Experimental data point to the possible therapeutic use of methylated lysine residues and methylated arginine residues in the case of neoplasms.

GnRH receptor and androgen receptor status and outcome of advanced prostate carcinomas.

BACKGROUND: The effect of gonadotropin-releasing hormone (GnRH) analogs on prostate carcinoma is partly a result of breaking the pituitary-gonadal axis, and partly the direct action on tumor cells expressing the GnRH receptor (GnRHR). The aim of this study was to detect the extent of correlation between the expression of GnRHR and also the androgen receptor (AR) and the efficacy of total androgen blockade (TAB). PATIENTS AND METHODS: Needle biopsy samples of twenty advanced prostate carcinoma patients were investigated histologically regarding Gleason score, AR and GnRHR status of the tumor cells. An affinity purified polyclonal antibody reacting with GnRHR and a monoclonal antibody for AR were applied for immunoperoxidase reactions. TAB was started in each case. Pathological, radiological and laboratory-prostate-specific antigen (PSA) data obtained before the start of TAB and survival, PSA values, and radiological findings after three years of TAB were related to AR and GnRHR. RESULTS: Regarding the clinical, radiological and laboratory findings before and after three years of TAB, 13 patients (group A) were considered to show a favourable' and 7 (group B) to show a 'poor' outcome. Twelve patients out of 14 with AR-positive tumor cells and all nine patients with GnRHR-positive tumor cells, as well as all eight patients with both AR- and GnRHR-positive tumor cells belonged to group A. The majority of AR-negative, GnRHR-negative or both AR- and GnRHR-negative cases belonged to group B. CONCLUSION: The presence of GnRHR and AR or both of these receptors indicates a more favourable outcome of advanced prostate carcinoma when treated with TAB, compared to GnRHR- and AR-negative cases. GnRHR and AR negativity may indicate a need for supplementary chemo- or radiotherapy.

Joint effects of cigarette smoking and irradiation in pregnant mice and their offspring.

BACKGROUND: We have previously reported that irradiation of mice in utero significantly increased the tumor incidence in the offspring of irradiated mothers. The joint effects of irradiation and cigarette smoking (CS) on tumor incidence and on the process of carcinogenesis were investigated. MATERIALS AND METHODS: Pregnant C57Bl/6J female mice were irradiated with a single dose of gamma-ray (1 Gy or 3 Gy) and/or exposed to CS of IR3 non-filtered cigarettes before or during pregnancy, tumors were investigated both with histological and immunohistochemical methods. RESULTS: Longer exposure (60 days) of the mice to CS before pregnancy and irradiation during pregnancy significantly increased the tumor incidence in the mothers and their offspring. Parallel activation of Caspase-8 and inactivation of Caspase-9 was found. CONCLUSION: Joint exposure of mice to prolonged CS before pregnancy and irradiation during pregnancy significantly increased the tumor incidence both in the mothers and their offspring.

Apoptosis regulation and spontaneous apoptosis index of testicular germ cell tumors are associated with differentiation and resistance to systemic treatment.

BACKGROUND: Cisplatin-based chemotherapy can cure more than 80% of metastatic germ cell testicular tumors (GCT). Germ cells are particularly susceptible to apoptosis and it is reasonable to presume that GCTs are curable because of an intact and effective apoptotic pathway. PATIENTS AND METHODS: The expression of p53 and p21 was investigated in conjunction with the spontaneous apoptotic index in 20 refractory and 50 chemosensitive GCTs, with a complete follow-up. To detect a differentiation-dependent alteration in the apoptotic pathway, all of the histological tumor types were examined separately. RESULTS: Embryonal carcinoma components showed significantly higher p53 expression compared to other histological subtypes of GCTs. p21 was barely detectable in the majority of tumors. Seminomatous components showed no p21 expression. Mature teratomas and syncytiotrophoblasts showed significantly higher p21 expression than other tumor subtypes. Embryonal carcinomas showed significantly higher apoptotic indices than other non-seminomatous components. On the other hand, choriocarcinomas and mature teratomas showed the lowest spontaneous apoptotic potential. The apoptotic index correlated with the fraction of p53-positive cells, but not with the p21 expression rate. The refractory group showed significantly lower p53 expression, higher p21 expression and a higher apoptosis index than the sensitive group. CONCLUSION: Our results suggest that the p53 and p21 expression levels and the apoptosis index seem to be important factors in the issue of the chemosensitivity of GCTs. The protein expression pattern reflects a differentiation-dependent preference for G1/S-phase arrest in terminally differentiated syncytiotrophoblasts and mature teratoma cells, while p53 mediated apoptosis induction is meaning to less differentiated tumor types.

Expression of p21(waf1/cip1), p27 (kip1), p63 and androgen receptor in low and high Gleason score prostate cancer.

The aim of this study was to investigate the expression of p21(waf1/cip1), p27(kip1), p63 and androgen receptor proteins in relation to serum prostate specific antigen levels in low and high Gleason score prostate cancers. Biopsies of patients suffering from prostate adenocarcinoma of low (3 + 3 to 3 + 4) and high (5 + 4 to 5 + 5) Gleason scores (13 cases each group) were immunostained for positive regulators of cell cycle control (p21(waf1/cip1) and p27(kip1)), and essential markers of normal prostate gland ontogeny (p63) and growth (androgen receptor) to find differentially expressed markers of malignant progression. Serum prostate specific antigen levels were also monitored at the time of biopsy and following anti-androgen therapy. All cases except one in each group were androgen receptor positive. P63 and p21(waf1/cip1) proteins detected in normal basal cell nuclei were lost in all but one studied tumors respectively. P27(kip1) protein, however, was detected in all low Gleason score prostate cancers, but it was found in only 7/13 high score cases. Prostate specific antigen levels, either pre- or post-treatment, did not show strict correlation with the p27(kip1) results. The low to high grade dedifferentiation of prostate adenocarcinoma is accompanied with the down-regulation of p27(kip1) protein, which may be an important molecular sign of the lost cell cycle control.

[Examination of somatostatin receptor expression in recurrent childhood medulloblastomas]. / A szomatosztatinreceptor-expresszió változásának vizsgálata recidiváló medulloblastomában.

Medulloblastoma is the most common malignant pediatric central nervous system tumor. Despite the adequate therapy the tumor often recurs. The primary medulloblastoma expresses somatostatin receptor-2 (SSTR-2), but so far we had no experience about the receptor status in recurrent tumors. The presence of SSTR-2 may have an important role in the early detection and treatment of recurrent medulloblastomas. Our aim was to examine the state of SSTR-2 expression in recurrent childhood medulloblastomas. We examined SSTR-2 expression by immunohistochemistry in primary and recurrent medulloblastoma samples of ten children treated with recurrent medulloblastoma at Semmelweis University, Departments of Pediatrics, between 1998 and 2004. All primary and recurrent tumors have been operated at the National Institute of Neurosurgery. We examined the intensity and the percentage of SSTR-2-positive tumor cells in the primary and recurrent tumor samples. All primary tumors were receptor-positive and SSTR-2 was also expressed in all recurrent medulloblastomas. In our samples the percentage of SSTR-2-positive tumor cells was 30-90%. As a positive in vivo control Octreoscan images were available in two cases. In these cases the results of immunohistochemistry and Octreoscan imaging seemed to correlate. As a conclusion, SSTR-2-positive recurrent tumors can be detected early by Octreoscan imaging, and the presence of SSTR-2 establishes the opportunity of applying somatostatin analogues (octreotide) in the treatment of recurrent childhood medulloblastoma.

Fermented wheat germ extract (avemar) inhibits adjuvant arthritis.

Anti-inflammatory efficacy of the fermented wheat germ extract (FWGE, Avemar) in the rat adjuvant arthritis (AA) model was examined. To Wistar rats with AA, different doses of FWGE and anti-inflammatory drugs (indomethacin, dexamethasone) as monotherapies were administered and FWGE and either diclofenac or dexamethasone were also given in combination. Besides plethysmographies of the paws, histological investigations of synovial tissues were also performed along with detection of CD4+ and CD8+ T lymphocytes. Gene expressions of COX-1 and 2 were determined by real-time polymerase chain reaction (PCR). FWGE monotherapy significantly inhibited the development of the secondary (immune-mediated) response in AA, and dexamethasone and indomethacin exerted inhibitory effects in a degree comparable to that of FWGE. Histological analysis of the affected joints confirmed the results. FWGE inhibited COX-1 and -2, while indomethacin enhanced COX-2 gene expressions. FWGE had an additive interaction with diclofenac. It is concluded that FWGE has significant anti-inflammatory efficacy confirmed by plethysmography, histology, and real-time PCR.

DNA and RNA isolated from tissues processed by microwave-accelerated apparatus MFX-800-3 are suitable for subsequent PCR and Q-RT-PCR amplification.

Over the past decade, methods of molecular biology have appeared in diagnostic pathology and are routinely applied on formalin-fixed, paraffin-embedded histological samples, processed via conventional embedding methods. Due to its reagent- and cost-effectiveness, embedding techniques that utilize microwave acceleration in one or more steps of histoprocessing are increasingly used by numerous laboratories. The demand arises that tissues processed this way should also be suitable for the requirements of molecular pathology. In this study, both conventionally embedded and MFX-800-3 machine-processed tissue samples from the same source were used for isolation of DNA and RNA and for performing PCR and real-time PCR. PCR amplification of the beta-globin gene, as well as the real-time PCR amplification of the ABL mRNA was successful in all cases. Our conclusion is that samples processed by the vacuum assisted automatic microwave histoprocessor MFX-800-3 are perfectly applicable for DNA and RNA isolation and provide appropriate templates for further PCR and realtime PCR studies.

Skull base chordoma mimicking a preauricular neoplasm in a child: clinicopathological features and biological behaviour.

INTRODUCTION: The extreme rarity of chordomas in childhood, the slow growing nature of these tumours and the diverse symptoms may cause many diagnostic problems. PATIENT: A 9-year-old girl presented with an unusual manifestation of a skull base chordoma. The clinical and pathological features were analysed. RESULT: In the present case, the initial symptoms of the skull base tumour were completely misleading. The otodynia, the masticatory difficulties and the mass in the preauricular region were not characteristic of skull base chordomas. The female sex, the young age, the large tumour size and the atypical histological pattern of the tumour all indicated a very poor prognosis. CONCLUSION: The rarity of this tumour in childhood and the atypical lateral and intracranial spread resulted in a serious delay of the diagnosis and in a fatal outcome.

The clinical and pathological symptoms of pregnancy epulis.

Examination of the oral cavity in 2260 pregnant women revealed 12 cases of pregnancy epulis, an incidence of 0.48%. In 9 of these 12 cases, spontaneous regression and recovery occurred 1-4 months after delivery. Despite the continuous conservative therapy, several surgical interventions were necessary in the remaining 3 pregnant women, due to the large size of the epulis and the bleeding that was difficult to stop. The authors provide a detailed description of the clinical symptoms (bleeding, difficulty of oral closure, loosening and migration of teeth) caused by large, multiple epulis instances during two pregnancies of one woman, together with the surgical interventions, the histopathologic examination and the immunohistochemical characteristics of the epulis.

Genotoxic potential of by-products in drinking water in relation to water disinfection: survey of pre-ozonated and post-chlorinated drinking water by Ames-test.

Mutagenic potential of drinking water samples derived from ranneywells was studied. 100-100 l of untreated (rough) and ozone-treated as well as chlorinated-disinfected water were dropped on and adsorbed by macroreticular resin columns (Serdolit PAD-III and Amberlite XAD-2). The adsorbed material was desorbed by methanol and dichloromethane. After elimination of the solvents by vacuum distillation the adsorbed material was dissolved in dimethylsulfoxide. The mutagenic activity was tested in the Ames-Salmonella/rat liver microsome system. The tester strains were TA-98 and TA-100. The material adsorbed to Serdolit PAD-III from rough and also disinfected water did not induce mutagenicity in case of the TA-98 tester strain, irrespective of activation by liver microsomes. However, the material adsorbed to Amberlite XAD-2 exerted mutagenic effect on the TA-98 tester strain, with and without liver microsome activation, both in case of rough and disinfected water. The TA-100 tester strain showed mutation after, but not without activation, when treated with the material adsorbed by either Serdolit PAD-III or Amberlite XAD-2, in case of rough water. Material derived from disinfected water and adsorbed to Serdolit PAD-III, caused mutation of the TA tester strain also only after activation. The material derived from disinfected water and adsorbed to Amberlite XAD-2 proved to be mutagenic to the TA-100 tester strain both without and after activation. Mutagenic activity was exerted by the amount of concentrates derived from 0.28 to 0.83 l of rough and 0.83-2.5l of disinfected water. The mutagenic activity of drinking water raises the possibility of carcinogenic effect, too. Search for alternative methods of water disinfection may be recommended.

[Changes in gene expression in the course of proliferative processes in the parathyroid gland]. / Génexpresszió-változások a mellékpajzsmirigy proliferatív folyamata során.

The aim of this study was to investigate the changes in expression pattern of the most important genes connected with apoptosis in proliferative apoptotic lesions (hyperplasia, adenoma), applying cDNA microarray technique, in order to promote the possible diagnostic or therapeutic utilisation of any difference in gene expression compared to the healthy (normal) parathyroid gland. Samples were taken from surgically removed 2 hyperplasias, 2 adenomas and 2 normal parathyroid glands. The Apoptosis Gene Array (Superarray) was used. This contains 112 genes, in tetraspot arrangement. The probes measured 250-600 base pairs. Streptavidin was bound to the array. CDP Star TM chemiluminescent substrate was used for detection. The samples deriving from hyperplasia or adenoma were compared to samples from normal parathyroid glands. The following genes were overexpressed in both hyperplasia and adenoma: CHEK1, ATM, BCL-XL, FAS, TNF, cIAP1, TRAIL, FADD, CASP 4,5,6,8, CD120b, CD137, LTA, TANK, TARF2, CAD, LIGHTR, DR3LG. CASP1,10, BFAR, BOD, BCL2L2, TRANCE were underexpressed in both hyperplasia and adenoma. Genes overexpressed only in hyperplasia were: MDM2, MCL1, BCL2A1, BLK, RIPK2, CD40LG, TRAF5, HUS1, BNIP3. Underexpressed only in hyperplasia: BOK, CIDEA, TRAF1, TRIP. Overexpressed only in adenoma: APOLLON, RIPK1, LTB, LTBR, CASP2,13, cIAP2, CIDEB. Underexpressed only in adenoma: TRAF4 and FASLG. Overexpresion or underexpression meant 1.5-fold difference from normal average values. As a result of this study, both pro-apoptotic and antiapoptotic genes were identified in hyperplasia and adenoma of the parathyroid gland. It seems that increased proliferation is connected also with increased apoptotic activity, but tumor cell candidates are able to survive, by activation of signal pathways resulting in overexpresion of anti-apoptotic genes.

Immunohistochemical demonstration of gonadotropin-releasing hormone receptors in prostate carcinoma.

The antiandrogen and gonadotropin-releasing hormone (Gn-RH) analogue treatment of prostate carcinoma is based on decreased proliferative and increased apoptotic activity of tumor cells, induced by androgen ablation. Gn-RH analogues decrease the serum level of androgens by breaking the pituitary-gonadal axis, but increasing evidence points to the direct effect of Gn-RH analogues on tumor cells. Immunohistochemical demonstration of Gn-RH receptors recently became possible. Cryostat and paraffin sections of prostate biopsy samples of 10 untreated patients with prostate carcinoma (T2-T3, Gleason score 5-8) were investigated for Gn-RH receptors (Gn-RHR) and androgen receptors, respectively, using the immunoperoxidase method. Membrane bound, focal Gn-RHR positivity was found in 5 samples. Nuclear androgen receptor positivity appeared in 7 samples. No correlation between Gn-RHR and androgen receptor positivity was found, neither receptor status and tumor-nodes-metastasis stage nor Gleason score could be related to each other. Correlation between Gn-RHR positivity and response to luteinizing hormone-releasing hormone analogue treatment will be investigated further.

Effect of Fraxiparine and heparin on experimental tumor metastasis in mice.

BACKGROUND: Low molecular weight heparins (LMWH) have become increasingly important in anticoagulant therapy. Antitumor and antimetastatic activity of heparin and LMWH-s have also been reported. MATERIALS AND METHODS: Fraxiparine, a new modified LMW-H, was tested for antimetastatic effect using 3LL-HH intravenous, B16 intra-foot pad and 3LL-HH intrasplenic models in C57 Bl/6 mice. The dose of Fraxiparine was 38, 57 and 172 IU/kg, respectively. Heparin (100 IU/kg) was used as a positive control. Both pre-treatment (starting 6 hours before tumor inoculation) and post-treatment (starting 24 hours after tumor inoculation), followed by daily injections, were applied in the intra-foot pad and intrasplenic models. In the intravenous model, only a single dose was administered one hour after tumor cell injection. RESULTS: Fraxiparine at the dose of 57 IU/kg was significantly antimetastatic in the intravenous model. Continuous treatment, starting 6 hours before tumor inoculation, with 173 IU/kg Fraxiparine resulted in a strong inhibition of lung metastases in the intra-foot pad model, but was ineffective in the intrasplenic model. Heparin did not influence the metastasis number in any of the metastasis models. CONCLUSION: These data may be of importance in the anticoagulant treatment of human cancer patients.

Development of present and future of telepathology in Hungary.

Pathologists in Hungary evaluate several hundred thousand histological specimens yearly, and a second opinion is requested in 5-10 percent of cases. Application of multimedia systems (i.e. telepathology) is convenient and efficient to establish a correct diagnosis in such cases. The first telepathology connection in Hungary has been established between the 1st Department of Pathology and Experimental Cancer Research, Semmelweis University and the Department of Pathology, Central Hospital of the Ministry of the Interior. Further development occurred in the course of various projects, supported by the EU (Inter-path, Re-transplant, Be Pro): new stations were established in three university institutes and six county hospitals. Electronic fixation of the images and their transmission by telephone line (ISDN) is easily available and an important feature of the multimedia system applied in telepathology. The system used by us is suited to evaluate frozen or paraffin-embedded histologic sections, as well as immunohistochemical and cytologic specimens, if necessary supplemented with transmission of macroscopic pictures. Our experience with bilateral consultations has proven the importance of telepathology. The telepathology system established in Hungary is now ready to join the telepathology network of the EU.

Proliferation and apoptosis in the corneal stroma in longterm follow-up after photorefractive keratectomy.

This study aimed at investigating the proliferation and apoptosis of corneal cells following photorefractive keratectomy (PRK) treatment. PRK (-6.0 D correction) was performed with the Asclepion-Meditec MEL70 G-scan excimer laser on the right eye of each of 33 rabbits under combined local and general anaesthesia. Animals were sacrificed at 4 h, 1, 4, 7, 14, 28, 56, and 112 days postoperatively, and corneal samples from these eight groups were examined histologically. Stromal cell proliferation was evaluated by immunocytochemical analysis of Ki67. Apoptosis was detected using the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling (TUNEL) assay method. The untreated left eyes served as controls. Ki67 positivity was detected in the upper stroma on day 1, 4, 7, and 14, and keratocyte apoptosis on day 1, 4, 7, and 14 after PRK, but not at an earlier or later time. Neither Ki67 positivity nor apoptotic activity was observed in the controls (untreated corneas). PRK was found to trigger proliferation and apoptosis of corneal keratocytes. The frequency and spatial distribution of keratocyte proliferation and apoptosis are likely to be important determinants of the corneal wound healing process, but the detailed regulatory mechanisms have not yet been characterized.

Chemopreventive properties of trans-resveratrol against the cytotoxicity of chloroacetanilide herbicides in vitro.

The beneficial effect of trans-resveratrol (RESV) on health is well documented. Our aim was to study the putative preventive effect of RESV on the cytotoxicity of frequently used herbicides (alachlor, acetochlor). Estrogen receptor positive (ER+) MCF-7 human mammary carcinoma, HepG2 (ER+) human hepatocellular carcinoma and VERO estrogen receptor negative (ER-) non-transformed monkey fibroblast cell lines were treated with alachlor and acetochlor (2-500 microg/ml) as toxic agents, and RESV (10 microM) as preventive agent. The MTT dye reduction assay was performed to test cytotoxicity, and flow cytometry to test cell proliferation and apoptosis. RESV is not cytotoxic in the concentration range of 1-100 microM on neither cell lines examined after 24 h, but cytotoxic on Vero and MCF-7 cells at 100 microM after 48h, and on all three cell lines after 72 h. On both ER+ cell lines a stimulation of viability occurs in the low concentration range (0.5-12.5 microM) as detected by the MTT assay. Cell cycle analysis of the culture shows a significant increase of S-phase cells at low concentrations of RESV (10-50 microM) and a decrease in the 100-200 microM concentration range. The ratio of apoptotic cells significantly increases after the administration of 50 microM RESV, depending on the incubation time. The cytotoxicity of 20-65 microg/ml alachlor and 10-65 microg/ml acetochlor was significantly decreased by the addition of 10 microM RESV in Vero ER- cells whereas no significant change was detected on ER+ cell lines MCF-7 and HepG2. These results show that RESV protects non-transformed ER- cells, but has no such effect on ER+ tumor cells.

Changes of protein expression in prostate cancer having lost its androgen sensitivity.

OBJECTIVE: The majority of prostate cancers require androgen hormones for growth, and androgen ablation is an important part of the systemic treatment of advanced prostate cancer. Nevertheless, most of these cancers eventually relapse as they become less sensitive to androgen ablation and anti-androgen treatment. Elucidating the molecular events that are responsible for the conversion of androgen-sensitive cancers to androgen-refractory tumors may reveal new therapeutic opportunities. METHODS: In the present study, we investigated nine androgen-sensitive and nine androgen-refractory prostate cancer samples to evaluate the expression levels of 10 selected proteins that have been implicated in oncogenesis and cancer progression. RESULTS: Our immunohistochemical data show that three of the investigated proteins (i.e., minichromosome maintenance-2, methylguanine-DNA methyltransferase, and androgen receptor) are expressed at significantly different levels in the androgen-refractory cancer samples than in the androgen-sensitive tumors, whereas the expression levels of the seven other studied proteins (i.e., ß-catenin, p27, p21, p16, Ki67, hypoxia-inducible factor 1 alpha, and geminin) are not significantly different regarding the two groups. CONCLUSIONS: Our data suggest that the increased expression of minichromosome maintenance-2 and decreased expression of methylguanine-DNA methyltransferase related to androgen receptor are indicative of the androgen-refractory stage in prostate cancer. Further studies are required to determine whether these expression changes play a causative role in the transition of androgen-sensitive to androgen-refractory prostate cancer.