A Study of Association between H. Pylori Genotype and Chronic Gastritis.

The prevalence of Helicobacter pylori (HP) infection is very high in Bangladesh. Chronic gastritis due to H. pylori is commonly associated with important gastric diseases such as peptic ulcer diseases or gastric carcinoma and MALT-oma. The natural course of chronic gastritis is HP-associated antral gastritis or pangastritis or rarely atrophies. This study was done to see the association of H. pylori genotypes with chronic gastritis. This observational cross sectional study was carried out at Bangabandhu Sheikh Mujib Medical University from July 2012 to April 2013 to find out the association of H. pylori genotypes with chronic gastritis in dyspeptic patients of Bangladesh. A total of 50 dyspeptic subjects were involved in the study whose upper GI endoscopies were carried out in presence of an experienced endoscopist. During the procedure four biopsies were taken, two from the antrum and two from body of the stomach. Endoscopic diagnosis was categorized into normal and erosive gastritis. Two (one from antrum and one from body) biopsy samples were collected in phosphate buffer saline and PCR analysis carried out by Multiplex PCR assay. Another two were collected in 10% formalin and histopathological examination was done according to updated Sydney system of classification. Among 50 patents only 34 were PCR positive. So, only 34 subjects were included in the study. Among them 21 patients (61.8%) were male and 13 patients (38.2%) were female, with the mean age of 29.91 years. Endoscopy revealed erosive gastritis in 5(14.7%) patients and normal findings in 29(86.3%) patients. Amongst the strains, cagA gene was detected in 58.8% and was not significantly associated with severity of any parameter of chronic gastritis such as H. pylori density, inflammation (mononuclear infiltration), activity (neutrophilic infiltration), atrophy and intestinal metaplasia. All the strains were positive for vacA allele. s1m1 (55.9%) genotype was most predominant. No vacA allele (s1m1, s1m2, s2m1 and s2m2) were significantly associated with severity of chronic gastritis. In this study, H. pylori genotype -cagA, vacA-s1, s2, m1, m2 allele and histological grading of chronic gastritis according to updated Sydney system of classification is identified. This study will identify the genotypes associated with severe gastritis in our country and thereby help us to take appropriate preventive measure. Further study with larger sample size may be carried out to establish proper association between different genotypes and parameters of chronic gastritis.

Role of cathepsin D in prostatic cancer cell growth and its regulation by brefeldin A.

We investigated a possible relationship between brefeldin A (BFA), an antibiotic, and cathepsin D (Cat.D), a lysosomal protease, in prostate cancer proliferation. Effects of BFA (30 ng/ml) were examined on the growth of three human prostatic cancer cell lines, PC-3, DU-145, and LNCaP cells. Its effect on Cat.D in these cancer cells was assessed by Western blots and compared with Cat.D expressed in clinical prostate specimens (n = 55). BFA profoundly (> 70%) inhibited the growth of all three cancer cell lines. Western blots revealed that expression of procathepsin D (Pro.Cat.D) was markedly increased with BFA, whereas actively proliferating (control) cells greatly exhibited mature Cat.D. Analysis of prostate specimens then showed predominant Pro.Cat.D expression in non-cancerous tissues while also showing enhanced expression of mature Cat.D in all cancer specimens. Therefore, BFA-induced growth inhibition in prostatic cancer cells is associated with a blocking of Cat.D maturation (activation), suggesting a possible role of Cat.D in prostate cancer proliferation/development.

Olanzapine versus haloperidol in children with autistic disorder: an open pilot study.

OBJECTIVES: Conventional neuroleptics ameliorate symptoms in children with autistic disorder; however, they are known to cause dyskinesias. Atypical neuroleptics, including olanzapine, may have less risk for dyskinesia, but their efficacy in autistic disorder is not established. This study was designed to investigate the safety and effectiveness of open-label olanzapine as a treatment for children with autistic disorder by using haloperidol as a standard comparator treatment. METHOD: In a parallel groups design, 12 children with DSM-IV autistic disorder (mean age 7.8+/-2.1 years) were randomized to 6 weeks of open treatment with olanzapine or haloperidol. Mean final dosages were 7.9+/-2.5 mg/day for olanzapine and 1.4+/-0.7 mg/day for haloperidol. Outcome measures included the Clinical Global Impressions (CGI) and the Children's Psychiatric Rating Scale (CPRS). RESULTS: Both groups had symptom reduction. Five of six in the olanzapine group and three of six in the haloperidol group were rated as responders according to the CGI Improvement item. Subjects showed improvement on the CPRS Autism Factor (F1,9 = 24.4, p = .0008). Side effects included drowsiness and weight gain. CONCLUSIONS: The findings suggest that olanzapine is a promising treatment for children with autistic disorder. Further placebo-controlled and long-term studies of olanzapine in autistic disorder are required.

Five-year prognosis after radical prostatectomy in a patient with localized prostate cancer and incidental non-Hodgkin's lymphoma.

We present a white male patient with an initial prostate-specific antigen level of 69 ng/ml, referred for urological evaluation. He was found to be free of prostatitis but diagnosed for prostate adenocarcinoma without any indications of metastatic disease. Lymphadenectomy then revealed lymphadenopathy of low-grade non-Hodgkin's lymphoma. Five-year follow-up after radical retropubic prostatectomy (RRP) showed no evidence of metastatic or local prostate cancer recurrence. In addition, no radiation or chemotherapy was required for his lymphoma. Although RRP is a viable option in this unique case, the outcome thus far suggests that it should be considered a primary therapeutic modality.

Glyoxalase I phenotype as a potential risk factor for prostate carcinoma.

OBJECTIVES: To elicit a possible link between glyoxalase I (Gly-I), a detoxifying enzyme, and the incidence of prostate cancer (PCa), we investigated Gly-I phenotypic expression in the prostatic tissue and red blood cells (RBCs) from patients with PCa. METHODS: Eighty-seven clinical specimens, including 42 PCa tissue samples, 20 RBC samples, and 25 matched pair (prostate and RBC) samples from patients at prostatectomy were examined. The Gly-I phenotypes in these specimens were assessed by nondenaturing starch-polyacrylamide gel electrophoresis. RESULTS: Of the 87 patients, 63 (72.4%) were white, 15 (17.2%) were black, and 9 (10.4%) were another ethnicity (eg, Hispanic, Asian, Indian). Three Gly-I phenotypes were detected in these specimens as fast, intermediate, and slow-moving bands on the gel. The fast phenotype was the most common form found in the white (34 [54%] of 63) and black (8 [53.3%] of 15) patients, but the third ethnic group was too small for proper analysis. To validate this finding, the data from the white patients were compared with the Gly-I phenotypic frequencies in U.S. populations. The data analysis confirmed that a higher incidence (54%) of the fast type in our white patients was statistically significant (P <0.0001) compared with its phenotypic frequency of 30.6% in the general U.S. white population. CONCLUSIONS: The significantly high frequency (P <0.0001) of the fast Gly-I phenotype was detected among patients with PCa, suggesting it is a potential risk factor for PCa. Whether its increased incidence in whites reflects the lack of sample numbers for other ethnic groups needs additional investigation.

Methylglyoxal-induced apoptosis in human prostate carcinoma: potential modality for prostate cancer treatment.

OBJECTIVE: To examine the cellular effects of methylglyoxal (MG), a toxic physiological metabolite, on human prostatic cancer PC-3 cells. METHODS: The effects of MG on cell growth and viability were evaluated first, and then its effects on the cell cycle and the glycolytic process were analyzed by Western blots and specific assays. Possible MG-induced apoptosis was also assessed by DNA analysis using agarose gel electrophoresis. RESULTS: MG > or =3 mM caused severe growth inhibition, resulting in nearly 100% cell death by 24h. The time course study revealed that expression of cyclin D(1), cdk2, and cdk4 was significantly (>50%) downregulated in 3 h of MG (3 mM) exposure, followed by the dephosphorylation of retinoblastoma protein by 6 h. Both the glyceraldehyde-3-phosphate dehydrogenase activity and the cellular lactate level were also reduced by approximately 50 and 80%, respectively, following 6-hour MG exposure. Induction of apoptosis by MG was indicated by partial degradation of poly(ADP-ribose) polymerase and further confirmed by discrete DNA fragmentation detected on an agarose gel. CONCLUSION: MG is capable of inducing apoptosis in prostatic cancer PC-3 cells, due primarily to a blocking of the cell cycle progression (G(1) arrest) and glycolytic pathway. Therefore, MG could be a potent apoptosis inducer, which may have a potential for prostate cancer treatment.

Induction of apoptosis in human prostatic cancer cells with beta-glucan (Maitake mushroom polysaccharide).

PURPOSE: To explore more effective treatment for hormone-refractory prostate cancer, we investigated the potential antitumor effect of beta-glucan, a polysaccharide of the Maitake mushroom, on prostatic cancer cells in vitro. MATERIALS AND METHODS: Human prostate cancer PC-3 cells were treated with various concentrations of the highly purified beta-glucan preparation Grifron-D(R) (GD), and viability was determined at 24 h. Lipid peroxidation (LPO) assay and in situ hybridization (ISH) were performed to unravel the antitumor mechanism of GD. RESULTS: A dose-response study showed that almost complete (>95%) cell death was attained in 24 h with GD > or = 480 microg/mL. Combinations of GD in a concentration as low as 30 to 60 microg/mL with 200 microM vitamin C were as effective as GD alone at 480 microg/mL, inducing >90% cytotoxic cell death. Simultaneous use with various anticancer drugs showed little potentiation of their efficacy except for the carmustine/GD combination (approximately 90% reduction in cell viability). The significantly (twofold) elevated LPO level and positive ISH staining of GD-treated cells indicated oxidative membrane damage resulting in apoptotic cell death. CONCLUSION: A bioactive beta-glucan from the Maitake mushroom has a cytotoxic effect, presumably through oxidative stress, on prostatic cancer cells in vitro, leading to apoptosis. Potentiation of GD action by vitamin C and the chemosensitizing effect of GD on carmustine may also have clinical implications. Therefore, this unique mushroom polysaccharide may have great a potential as an alternative therapeutic modality for prostate cancer.

Ammonium-chloride-induced prostatic hypertrophy in vitro: urinary ammonia as a potential risk factor for benign prostatic hyperplasia.

To test the possibility that urinary ammonia could be a risk factor for benign prostatic hyperplasia (BPH), we explored the cellular effects of ammonium chloride (NH(4)Cl) on prostatic cancer cells used as an experimental model. Following treatment of human prostatic cancer DU-145 cells with the varying concentrations of NH(4)Cl for 3 days, cell growth was inhibited by approximately 50% at 5 mM NH(4)Cl and almost completely inhibited at 10 mM NH(4)Cl. However, the individual cell size in these treated cells became approximately 2-fold larger and cellular protein content was also up to 2.5-fold greater than in untreated cells. This protein increase appeared to result from the reduced protein degradation, verified by metabolic labeling with [(14)C]valine. Western blot analysis further suggested that such reduced protein turnover could in part be due to the inactivation of a lysosomal acid protease, cathepsin D. Taken together, these studies demonstrate NH(4)Cl-induced hypertrophy in prostatic cancer cells, as evidenced by the growth inhibition, cell enlargement, and cellular protein increase. Therefore, ammonia is not an inert metabolic product; instead, its chronic effects on the prostate may ultimately lead to significant cellular and biochemical alterations of the prostate such as BPH.

Glyoxalase I activity in human prostate cancer: a potential marker and importance in chemotherapy.

PURPOSE: To provide information on the activity of Gly-I in prostate cancer. MATERIALS AND METHODS: We performed qualitative Gly-I assay on prostate tissues. RESULTS: Gly-I activity between prostate cancer and noncancerous specimens differed substantially and significantly, although such activity also varied somewhat among cancer specimens. CONCLUSIONS: Gly-I activity is indeed higher in cancerous than in noncancerous specimens, suggesting that it may play a role in prostate cancer homeostasis and survival.

Analysis of cathepsin D forms and their clinical implications in human prostate cancer.

PURPOSE: To assess cathepsin D (Cat.D) status in the prostate, we analyzed the different Cat.D forms in human prostate tissues using Western immunoblots. MATERIALS AND METHODS: Cell extracts were prepared from prostate tissues (n = 42) obtained from radical prostatectomy, adopting the tissue homogenization method. Expression of the different Cat.D forms was analyzed using Western blots. The catalytic activity of Cat.D was assayed by acid treatment, in which cell extracts were incubated in acidic buffer (pH 3 to 4) at 37C for 1 hour. RESULTS: Pathologically confirmed normal (NML), benign prostatic hyperplasia (BPH) and cancer (CAP) specimens all expressed Cat.D, but as two distinct forms. Both NML and BPH predominantly expressed an inactive procathepsin D (Pro.Cat.D), while CAP notably exhibited an active mature Cat.D. The assessment of Cat.D activity, using PSA (prostate specific antigen) as a physiological substrate, showed that such activity was consistently higher in CAP than in NML/BPH specimens. Further studies revealed that the mode of Cat.D activation in CAP specimens appeared to be primarily due to acid-induced autoproteolysis (self-degradation) of mature Cat.D. CONCLUSION: This study demonstrates that expression and activity of Cat.D varies among prostate specimens. A greater expression of mature Cat.D with a higher catalytic activity in CAP specimens is the most notable difference from NML/BPH. Therefore, the differential expression/activity of Cat.D forms may be a useful indicator for assessing prostate cancer status.

Hydrolysis of androgen receptor by cathepsin D: its biological significance in human prostate cancer.

OBJECTIVE: To elicit the biological role of a lysosomal protease, cathepsin D (CatD) in prostate cancer, by investigating its regulatory effect on the androgen receptor (AR) using human prostate cancer LNCaP cells and prostate tissue specimens. MATERIALS AND METHODS: Cell extracts were prepared from LNCaP or prostate specimens by cell lysis and tissue homogenization. Proteolytic assays were performed by incubating these extracts in acidic buffer (pH 3-4) at 37 degrees C. The resulting effects on AR and CatD were then analysed using Western immunoblots. RESULTS: The Western blots showed that AR was virtually hydrolysed with acid treatment, because endogenous CatD was activated; this activation only occurred at pH 3.2-3.5, but no specific acid appeared to be required. Further analyses suggested that CatD activation could be attributed to acid-induced autoproteolysis of mature CatD. Similar assays were also performed on prostate tissues, including normal and malignant specimens. These studies revealed that CatD-mediated AR hydrolysis was observed only in cancer specimens, while no such hydrolysis occurred in normal specimens. CONCLUSION: Endogenous CatD can hydrolyse AR, thereby possibly modulating AR function/metabolism in LNCaP cells, and in cancer specimens. CatD activity also appears to differ significantly between normal and malignant tissue. Thus, CatD may play a pivotal role as a growth modulator in androgen-dependent prostate cancer.

Comparison of fluorescence in situ hybridization and flow cytometric DNA ploidy analysis in paraffin-embedded prostatic adenocarcinoma specimens.

Cancer cell nuclei extracted from 15 paraffin-embedded radical prostatectomy specimens were examined with fluorescent in situ hybridization (FISH) and flow cytometry (FCM) for DNA ploidy. Repetitive chromosome-specific alpha-satellite DNA probes to centromeres 7 and 10 were used in the FISH assay. Results from FISH and FCM were compared and viewed in relation to clinical experience. Of the 15 tumors examined, 9 were hyperdiploid for at least one chromosome by FISH assay. Seven were aneuploid by FCM, showing a good correlation between the two methods (P = .006). Using either evidence of clinical progression or a postoperative prostate-specific antigen (PSA) level > 0.5 ng/mL by Hybritech assay to indicate the risk of clinical progression, 56% of FISH hyperdiploid tumors had a risk of progression of carcinoma versus 17% of FISH diploid tumors. A preoperative PSA level > 30 ng/mL had a marginal correlation with risk of progression (P = .05). We demonstrate that FISH is a useful tool for chromosomal analysis in paraffin-embedded tumor specimens.

Urologic complications of high-dose chemotherapy and bone marrow transplantation.

OBJECTIVE: This study reviews the incidence and management of cyclophosphamide-induced hemorrhagic cystitis in a group of patients who received high-dose chemotherapy and bone marrow transplantation. METHODS: The records of 217 consecutive patients undergoing bone marrow transplantation were reviewed. The incidence, degree, and management of hematuria in this group of pancytopenic and immunocompromised patients were recorded. RESULTS: Despite prophylaxis, cystitis developed in 58 of these 217 patients (27%). In 12 patients (6%) the cystitis was severe. These patients had gross hematuria, clot retention, and drop in hematocrit necessitating blood transfusion. These patients were managed with continuous bladder irrigation, alum irrigation, and when less aggressive approach was unsuccessful, with intravesical formalin instillation. Alum irrigation was used in 5 patients, and was successful in only 1 patient. Six patients required intravesical formalin instillation to control the hematuria. Formalin solution 2-5% was instilled initially. When lower-concentration formalin failed, 5-10% formalin was used progressively. CONCLUSIONS: Patients with bone marrow transplantation in whom severe hemorrhagic cystitis develops should be managed aggressively early. Intravesical formalin appears to be the most effective regimen in controlling profuse, persistent hematuria.

Squamous cell carcinoma arising from suprapubic cystotomy site without bladder involvement.

We report on an eighty-year-old Haitian man with a suprapubic mass of seven months duration after five years of urinary diversion for urethral stricture. Histologically the mass was a squamous cell carcinoma confined to the suprapubic tract without bladder involvement. We believe this is the first such case reported in the literature, and it stresses the need for close monitoring of patients with any type of long-term indwelling catheter.

New Cavitron system (CUSA/CEM): its application for kidney surgery.

A new Cavitron Ultrasonic Surgical Aspirator system (CUSA/CEM), which contains electrocautery function at the hand piece, was used for partial nephrectomies in dogs without clamping the renal artery. The advantages of this new device (CUSA/CEM) over the original CUSA are: (1) it enabled more precise resection of the tissue in a dry field, (2) shorter operating time, and eventually (3) helpful for reduction of blood loss. The microscopic examination of the resected kidney revealed minimal reaction and damage to the adjacent kidney tissue as compared with the effect by electrocautery or microwave tissue coagulator.

Maintenance intravesical bacillus calmette-guerin for superficial transitional cell carcinoma of bladder. A retrospective study.

Eighteen patients with recurrent and/or multifocal superficial transitional cell carcinoma of the bladder who were rendered tumor-free by transurethral resection and were then treated with either a single or second six-week course of induction Bacillus Calmette-Guerin (BCG) therapy, followed by maintenance therapy, were retrospectively reviewed. A 73 percent complete response rate was achieved in those patients treated prophylactically, while a 70 percent complete response rate was observed in patients treated for carcinoma in situ (CIS) with an average follow-up of twenty-nine months. Maintenance therapy may be warranted in those patients able to tolerate it without significant side effects.

Establishment of new human prostatic cancer cell line (JCA-1).

The establishment of a new human prostatic cancer cell line is described. This cell line was derived from a poorly to moderately differentiated prostatic adenocarcinoma. It has been maintained in tissue culture for fourteen months and has been passed fifty-two times. This cell line has an ability to form colonies in soft agar suspension cultures, and also is transplantable to nude mice. Tumors grown in nude mice revealed a poorly differentiated adenocarcinoma with positive PSA staining. Acid phosphatase activity was detected in freeze-thawed cells by enzymatic assay. A karyotype analysis demonstrated aneuploidy with a model chromosomal number of 69 and six marker chromosomes.

Correlation of heterogeneity index score (HIS) of DNA content in bladder cancer recurrence.

Automated flow cytometry (FCM) has been used to monitor the effects of therapy and progression of human bladder carcinoma. We have previously reported a computer-based model which has shown a correlation to relative mean DNA content in cell populations analyzed by FCM. Two patients with superficial transitional cell carcinoma of the bladder were followed for several months. Using FCM the patients' tissue samples were examined and heterogeneity index scores (HIS) were determined. Both cases had recurrence. The first patient has increased in grade with a respective increase in HIS within four months (grade I----II, HIS 27.2----80.8). The second patient also has recurrence in three months both times with a grade I tumor and a score of 106.2. High scores reflect large aneuploid populations which have shown to recur. Since such a correlation exists HIS may not only offer a more objective technique with quantitative results to monitor patients but possibly can distinguish the degree of tumor malignancy.

Cavitrons in urologic surgery.

The CUSA is a unique modality for precise removal of tissue. This ultrasonic scalpel represents a major technical advance in surgery by providing a small handpiece device that can selectively remove tissue in a controlled manner. The usefulness of this modality in removing parenchymal tissue while sparing vascular or ductal structures is outstanding. Laboratory and clinical experience have demonstrated that utilization of this scalpel, particularly in vascular organs and lesions, provides the surgeon with increased visibility, reduced bleeding, and shorter operating time. Our application of this instrument in the performance of renal surgery in dogs as well as in the clinical setting has been well documented. Its future application as a surgical tool in the armamentarium of the urologist is uncertain and awaits the outcome of future studies.

Cavitron ultrasonic surgical aspirator. Applications in urologic surgery.

The Cavitron Ultrasonic Surgical Aspirator is a unique modality for precise tissue removal which results in increased visibility, reduced bleeding, and shorter operating time when dealing with vascular lesions or organs. Herein, we present our experience in a dog laboratory as well as in a clinical setting with this new instrument. The mechanism of action, proper surgical technique required, and further application in urologic surgery are discussed.