Evaluation of cytotoxic and anticancer effect of Orobanche crenata methanolic extract on cancer cell lines.

We aimed to assess the antitumor activity of Orobanche crenata methanolic extract and evaluate its cytotoxic effect on different cancer cell lines to develop an effective natural anticancer drug. Components of O. crenata methanolic extract were analyzed using gas chromatography-mass spectrometry. The extract's antioxidant activity was assessed by 2,2-diphenyl-1-picrylhydrazyl and ferric reducing antioxidant power procedures and cytotoxicity of the extract was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase assays. Caspase-3 activity was also estimated. O. crenata methanolic extract shows powerful antioxidant activity. The extract inhibited the propagation of human hepatocellular carcinoma (HepG2), human prostate cancer (PC3), human breast adenocarcinoma (MCF-7), and human colon carcinoma (HCT-116) in a dose-dependent manner. O. crenata-treated cells displayed obvious morphological structures distinctive of apoptosis. MTT assay exposed that the extract presented prevention of cell persistence in a dose-dependent means and revealed extremely cytotoxic activity against HepG2, PC3, MCF-7, and HCT-116 with 50% inhibitory concentration values 30.3, 111, 89.6, and 28.6 µg/mL, respectively, after 24 h of incubation. In addition, treatment of HCT-116 with various concentrations of the extract caused the release of lactate dehydrogenase and induction of caspase-3 activity in a dose-dependent way. In conclusion, our findings suggested that the O. crenata extract possesses potent antioxidant, cytotoxic activity, and anticancer properties which are possibly due to the principal bioactive phytochemical composites existing in this plant. These results can be used to develop new drugs for cancer treatment.

Evaluation of Osteopontin and Pokémon genes expression in hepatitis C virus-associated hepatocellular carcinoma.

Osteopontin and Pokémon genes may have an important role in the pathogenesis of different malignancies. Osteopontin is a glycoprotein of the extracellular matrix, and Pokémon is a regulator of transcription. Both have been hypothesized to be useful as therapeutic targets or diagnostic markers. We aim to assess the role of both in hepatocellular carcinoma and liver fibrosis due to hepatitis C virus (HCV) infection. We conducted our study on 50 patients and classified them into three groups-Group I: Patients with HCV-related hepatocellular carcinoma (HCC) (n = 30); Group II: Patients with hepatitis C cirrhosis (n = 10); and Group III: Patients with hepatitis C fibrosis (n = 10). We found high levels of Osteopontin and Pokémon gene expression in group I. Osteopontin levels were higher also in patients with liver fibrosis was correlated to high levels of parameters such as alpha fetoprotein and caspase. We conclude that HCC is associated with overexpression of both Osteopontin and Pokémon and that Osteopontin plays a significant role in liver fibrosis due to hepatitis C infection.

Biological activity of Echinops spinosus on inhibition of paracetamol-induced renal inflammation.

This study was designed to evaluate the possible mechanisms through which Echinops spinosus (ES) extract demonstrates nephroprotective effect on the paracetamol acetominophen (N-acetyl-p-aminophenol (APAP)) induced nephrotoxicity in rats. Twenty-four Swiss albino rats were divided into four groups (six rats each). The placebo group was orally administered sterile saline, the APAP group received APAP (200 mg·kg-1·day-1 i.p.) daily, the ES group was given ES extract orally (250 mg/kg), and the APAP + ES group received APAP as for the APAP group and administrated the ES extract as for the ES group. Pretreatment of methyl alcohol extract of ES reduced the protein expression of inflammatory parameters including cyclooxygenase-2 and nuclear factor κB in the kidney. It also reduced the mRNA gene expression of tumor necrosis factor-α and interleukin-1ß. The ES extract compensated for deficits in the total antioxidant activity, suppressed lipid peroxidation, and amended the APAP-induced histopathological kidney alterations. Moreover, ES treatment restored the elevated levels of urea nitrogen in the blood and creatinine in the serum by APAP. The ES extract attenuated the APAP-induced elevations in renal nitric oxide levels. We clarified that the ES extract has the potential to defend the kidney from APAP-induced inflammation, and the protection mechanism might be through decreasing oxidative stress and regulating the inflammatory signaling pathway through modulating key signaling inflammatory biomarkers.

Assessment of anticancer activity of Pulicaria undulata on hepatocellular carcinoma HepG2 cell line.

Searching for new sources of safe nutraceuticals antitumor drugs is an important issue. Consequentially, this study designed to assess the antitumor activity of Pulicaria undulata extract in vitro in the treatment of hepatocellular carcinoma HepG2 cell line. Aerial parts of P. undulata plants were collected, used for phytochemical analysis, and assessed for anticancer activity. The antitumor activity was evaluated through studying the cell viability and apoptotic pathway. The gas chromatography-mass spectrometry phytochemical analysis revealed that P. undulata is a promising new source of several known antioxidant and antitumor compounds which could participate in drug development and exploration of alternative strategies to the harmful synthetic antitumor drugs. P. undulata stifled HepG2 cell viability in a concentration-dependent manner. Meanwhile, P. undulata tempted substantial apoptosis in HepG2 cells and enhanced the expression of miR-34a. However, the mRNA expression level of antiapoptotic B-cell lymphoma-2 was markedly decreased by P. undulata treatment. Moreover, P. undulata increased the protein expression of proapoptotic p53 and caspase 3/9 with reducing B-cell lymphoma-2 protein expression level. Thus, P. undulata induced apoptosis in the HepG2 cells by overexpression of miR-34a which regulates p53/B-cell lymphoma-2/caspases signaling pathway. These findings were well appreciated with morphological studies of cells treated with P. undulata. In conclusion, P. undulata could be a probable candidate agent for the initiation of cell apoptosis in HepG2 and thereby can serve as promising therapeutic agent for treatment of hepatocellular carcinoma which should attract further studies.

Diabetic Nephropathy Assessment: Microtubule-Associated Protein 1 Light-Chain 3B a New Promising Biomarker.

Autophagy is a highly conserved pathway. Impairment of autophagy is implicated in the pathogenesis of diabetic nephropathy. The current study applied a bioinformatics analysis to retrieve promising autophagy biomarker relevant diabetic nephropathy. Urinary expression of Microtubule-associated protein 1 light-chain 3B (LC3B) RNA was assessed. Urine samples of 86 type II diabetic kidney disease Egyptian patients (albuminuria group) were provided to quantify urinary expression of LC3B. A group of 30 healthy volunteers were also enrolled in addition to non-albuminuria group including 44 patients. Our study revealed a cut-off value for urinary LC3B expression level that was calculated by receiver-operating characteristic curve as 0.866. Sensitivity and specificity of LC3B were 83.7 and 78.4% respectively. The positivity rate of urinary LC3B expression level was significantly lower in diabetic nephropathy patients than control group. LC3B has great clinical value as promising biomarker in diabetic nephropathy assessment.

Caffeic Acid Modulates miR-636 Expression in Diabetic Nephropathy Rats.

We investigated the action of caffeic acid in regulating miR-636 expression level in kidney of streptozotocin-induced diabetic rats. Streptozotocin-induced diabetic rats were orally treated with caffeic acid at 40 mg/kg/day for 8 weeks. At the end of the treatment, body and kidney weight and blood glucose levels were determined, blood, urine, and kidneys were collected for biochemical and histological examination. Expression levels of miR-636 were determined in liver by qRT-PCR. Induction of diabetic nephropathy by streptozotocin was evidenced by displayed elevated levels of serum creatinine, blood urea nitrogen, microalbuminuria and urinary albumin/creatinine ratio in addition to renal hypotrophy. Caffeic acid (CA) can ameliorate renal damage and significantly decreased the fasting blood glucose, cholesterol and triglyceride in diabetic rats. CA treatment improved histological architecture in the diabetic kidney. CA significantly down regulate miR-636 expression level in the kidney of diabetic rats in comparison to healthy group. Overall, caffeic acid down regulates miR-636 expression level which is involved in development of diabetic nephropathy and might therefore be potential attractive therapeutic agent to pursue in DN.

Possible Role of microRNA-122 in Modulating Multidrug Resistance of Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is a hypervascular primary liver cancer characterized by rapid progression, besides, resistance to traditional chemotherapeutic agents. It has been shown that microRNAs play critical roles in regulation of tumor cell sensitivity to drugs through modulating the expression of genes involved in drug transport. The present study investigated whether restoration of miR-122 in HCC cells could alter the cell cycle distribution and the expression of multidrug resistance (MDR)-related genes (ABCB1, ABCC1, ABCG2 and ABCF2). After overexpression of miR-122 in HepG2 cells treated or untreated with doxorubicin doses, total RNAs and protein extracts were isolated for application of QRT-PCR and western blotting techniques. Moreover, cell cycle distribution was monitored by flow cytometry. Our results revealed that, the over expression of miR-122 in HepG2 cells treated or untreated with doxorubicin could modulate the sensitivity of cells to chemotherapeutic drug through downregulation of MDR-related genes, ABCB1 and ABCF2. Interpretation of cell cycle distribution revealed that, the anti-proliferative effect of miR-122 is associated with the accumulation of cells in G0/G1 phase. Moreover, treatment with miR-122 and doxorubicin resulted in high percentage of HCC cells in G0/G1 phase. Taken together, our findings revealed that, overexpression of miR-122 inhibited HCC cell growth by inducing cell cycle arrest and this arrest is associated with down-regulation of MDR-related genes.

Oxidant/antioxidant status in children and adolescents with immune thrombocytopenia (ITP) and the role of an adjuvant antioxidant therapy.

BACKGROUND: Free oxygen radicals might have an adverse effect on platelets which might be reflected either on its count and/or degree of bleeding severity. AIM: To assess oxidant-antioxidant systems and evaluate effect of antioxidant therapy on platelet count (PC) and bleeding score (BS) in children and adolescents with ITP. METHODS: Six months prospective randomized single blind study registered as (NCT 01763658) including 39 patients with newly diagnosed (ND) ITP; group 1 (G1) and 39 patients with chronic ITP (G2), each group was randomly allocated (2:1) to one of two subgroups respectively; (G1A and G2A) interventional arm received daily antioxidant therapy, while G1B and G2B; received a placebo. Both groups were compared with healthy controls (n = 39). The primary efficacy endpoints were the difference in the change from baseline to 6 month in ITP specific bleeding assessment tool (ITP-BAT), PC, total antioxidant capacity (TAC), catalase (CAT), reduced glutathione (GSH) and serum malondialdehyde (MDA). RESULTS: Baseline TAC was significantly lower in patients with (ND) ITP compared to patients with chronic ITP (P < 0.05), both showed significantly lower levels than healthy controls (P < 0.001). At end of study both BS and PC significantly improved in patients receiving antioxidant compared to placebo (P < 0.05). Patients with chronic ITP receiving antioxidant showed better improvement. CONCLUSION: Reduced antioxidant mechanisms were reported in patients with ITP. Antioxidant therapy ameliorated the oxidative stress in both ND and chronic ITP groups which might explain the improvement in both BS and PC.

Ethanolic extract of Trigonella Foenum Graecum attenuates cisplatin-induced nephro- and hepatotoxicities in rats.

Nephro-and hepatotoxicities are important complications in cancer patients undergoing cisplatin (CP) therapy. We aimed to study the protective effect of fenugreek (FG) on CP induced renal and hepatic injuries in rats. Cisplatin intoxication resulted in structural and functional renal and hepatic impairments, which were revealed by massive histopathological changes and elevated kidney and liver function tests. However, it was associated with oxidative stress and lipid peroxidation as evident by increased reactive oxygen species (ROS) and malondialdehyde (MDA) with decreased levels of total antioxidant activity. Cisplatin administration triggered inflammatory responses and apoptosis in rat livers and kidneys as evident by increased expression of pro-inflammatory cytokine, tumor necrosis factor- α (TNF-α) and apoptotic marker p38 mitogen-activated protein kinase (p38 MAPK) as results of overproduction of ROS. FG significantly attenuated the cisplatin-induced biochemical and histopathological alterations, inflammation and apoptosis in rat livers and kidneys. Results suggested that fenugreek co-administration has a powerful antioxidant effect and may serves as a novel and promising preventive strategy against cisplatin-induced nephron- and hepatotoxicities.

Value of vanin-1 assessment in adult patients with primary immune thrombocytopenia.

The diagnosis of primary immune thrombocytopenia (ITP) is clinical and cannot be established by any specific laboratory assay. Perhaps the best diagnostic study is assessment of the patient's response to ITP therapy. Oxidative stress-related pathways were among the most significant chronic ITP-associated pathways. Overexpression of VNN1 gene, an oxidative stress sensor in epithelial cells, was most strongly associated with progression to chronic ITP. To address this issue, we tested the hypothesis that blood vanin-1 protein level could distinguish between chronic responders and non-responders ITP patients as well as between ITP patients and healthy controls. Vanin-1 protein levels were determined in peripheral blood leukocytes of 80 adult subjects (16 newly diagnosed ITP patients, 24 chronic responders ITP patients, 24 chronic non-responders ITP patients and 16 healthy controls) by enzyme-linked immunesorbent assay (ELISA). Blood vanin-1 protein levels were lower in controls (median = 18.39 ng) than in ITP patients (median = 58.78 ng) with a highly significant p value (p < 0.001). Vanin-1 levels were highly significantly elevated in newly diagnosed ITP patients (median = 188.62 ng) in comparison to chronic responders (median= 26.90 ng) and chronic non-responders (median = 73.87 ng). Vanin-1 level at a cut-off value of >20.73 ng was found to be 100% sensitive and 93.7% specific in discriminating between newly diagnosed ITP patients and healthy controls. Vanin-1 level was found to be 100% sensitive and 100% specific in differentiating between responders and non-responders with a cut-off value of ≤ 34.5 ng. Our results suggest that vanin-1 can distinguish between chronic responders and non-responders ITP patients as well as between newly diagnosed ITP patients and healthy controls. These findings demonstrate that vanin-1 may contribute to the pathogenesis of ITP, indicating that vanin-1 is an important target for further investigation.

Potential diagnostic role of circulating MiRNAs in colorectal cancer.

OBJECTIVES: Colorectal cancer (CRC) is the third most common and fourth most deadly cancer worldwide despite its various screening method. Thus, the search for novel and better markers is continuous. This study aimed to assess the combined expression levels of miR-133a, miR-574-3p, and miR-27a in early diagnosis of colorectal cancer in comparison to traditional tumor markers (CEA and CA19.9). METHODS: miR-133a, miR-574-3p, and miR-27a were assessed in sera of 120 participants categorized into healthy control group (n = 20), benign group (n = 30) and malignant group (n = 70) using real-time PCR. RESULTS: miR-133a, miR-574-3p, and miR-27a expressions showed significant difference among different staging, grading and tumor size of CRC. The sensitivities of the three miRNAs whether combined or individually used were better than routinely used tumor markers (CEA and CA19.9) leading to more accurate and faster diagnosis of CRC. CONCLUSION: Synergetic detection of miRNA-133a, miRNA-574-3p, and miRNA-27a may serve as better noninvasive biomarkers with higher combined sensitivity for early diagnosis of CRC than individual detection of miRNAs.

Evaluation of Brown Micro-Algae Synergies With Low Dose γ-Radiation Against Chronic Hepatitis Induced by D-Galactosamine in Rats.

Introduction: Hepatic inflammation is considered key driver of hepatic tissue impairment.We aimed to explore the interaction of Halamphora coffeaeformis (Amph.) with low dose ionizing γ radiation (γR) exposure against D-galactosamine (D-GaIN)-induced chronic hepatitis in Albino rats. Methods: Chronic hepatitis was induced with single dose of D-GalN (400 mg/kg BW i.p.). Rats received 400 mg Amph/kg BW daily by gastric gavage concomitant with .25 Gy γ-R. Liver oxidative stress and inflammatory status were assessed. Gene expression levels of signal transducer and activator of transcription 3 (STAT3) and nuclear factor kappa B (NFKB) were estimated by q-PCR. D-Galactosamine injection significantly encouraged hepatic oxidative damage and inflammatory disturbance accompanied with improved intercellular adhesion molecule-1 level (ICAM-1). Results: messenger RNA gene expression levels of STAT3 and NF-kB were expressively higher in D-GaIN-treated animals. Histopathological examination supported results. Interestingly, Amph treatment with γ-radiation (γ-R) subjection displayed significant improvement of oxidative and inflammatory status along with controlled signaling molecular factors which was supported by amended histological structure of induced liver hepatitis. Conclusion: Results conclude the efficacious control of liver hepatitis progression by dual collaboration of Amph. with low dose γ-R via control of vital growth signaling factors linked with inflammation thru anti-inflammation, antioxidative and anti-proliferative activities.

Ameliorative potential of manganese nanoparticles with low-level ionizing radiation against experimentally induced hepatocarcinogenesis.

Nanotechnology is a rich field with infinite possibilities of drug designs for cancer treatment. We aimed to biosynthesize manganese nanoparticles (Mn NPs) using Lactobacillus helveticus to investigate its anticancer synergistic effect with low-dose gamma radiation on HCC-induced rats. Diethylnitrosamine (DEN) (20 mg/kg BW, 5 times a week for 6 weeks) induced HCC in rats. Rats received Mn NPs (5 mg/kg BW/day) by gastric gavage over 4 weeks concomitant with single dose of gamma radiation (γ-R) (0.25 Gy). Characterization, cytotoxicity, and anticancer activity of Mn NPs were evaluated. DEN-induced significant liver dysfunction (alanine transaminase activity ALT, total proteins, and albumin levels) associated with significant increase in lipid peroxidation levels with reduction in super oxide dismutase activity. Furthermore, DEN intoxication is sponsored for remarkable increase in levels of Alfa-fetoprotein, tumor necrosis factor α, vascular endothelial growth factor, and transforming growth factor beta with remarkable decrease in caspase 3 and cytochrome c. Treatment with Mn NPs (4.98-11.58 nm) and single dose gamma radiation evoked significant repair in ALT, total protein, and albumin accompanied with balanced oxidative status, diminished inflammatory biomarkers, angiogenic factor, and growth factor with restoration in apoptotic factors. Mn NPs revealed obvious in vitro cytotoxic activity against HepG2 cell line in a dose-dependent manner. Our findings were well appreciated with the histopathological study. In conclusion, a new approach of the single or combined use of Mn NPs with low-dose γ-radiation regimens as promising paradigm for HCC treatment is recommended.

Curative role of mesenchymal stromal cells in chronic pancreatitis: Modulation of MAPK and TGF-ß1/SMAD factors.

BACKGROUND AND OBJECTIVE: Living organisms respond to physical, chemical, and biological threats with a potent inflammatory response which alters organ cell signaling and leads to dysfunction. We evaluated the therapeutic effect of bone marrow-based mesenchymal stromal cell (BM-MSC) transplanted in rats to preserve tissue integrity and to restore homeostasis and function in the pancreatitis experimental pattern. METHODS: This study involved 40 adult male Wister rats. Repeated L-arginine injections caused chronic pancreatitis (CP), leading to the development of pancreatic damage and shifting the intracellular signaling pathways. Rats were then infused with BM-MSC labeled with PKH26 fluorescent linker dye for 12 weeks. RESULTS: Cell-surface indicators of BM-MSCs such as CD 90 and CD29 were expressed with the lack of CD34 expression. BM-MSC treatment considerably improved the alterations induced in a series of inflammatory markers, including IL-18, TNF-α, CRP, PGE2, and MCP-1. Furthermore, improvement was found in digestive enzymes and lipid profile with amelioration in myeloperoxidase activity. BM-MSC treatment also regulated the (TGF-/p-38MPAK/SMAD2/3) signaling factors that enhances repair of damaged pancreatic tissue, confirmed by reversed alteration of histopathological examination. CONCLUSION: our results further bring to light the promise of cell transplant therapy for chronic pancreatitis.

Assessment of the association of serum progranulin with autophagy in diabetic patients.

INTRODUCTION: Progranulin (PGRN) has newly arisen as an important regulatory protein of glucose metabolism and insulin sensitivity. Progranulin expression is interrelated with lysosomal function strongly linked to autophagy pathway. We aimed to evaluate the correlation between PGRN protein and microtubule-associated protein light chain 3B (LC3B) expression level in diabetic patients. MATERIAL AND METHODS: Blood samples of 70 type 2 diabetic Egyptian patients were provided for analysis of concentrations of serum progranulin and interleukin 6 (IL-6) using ELISA, and quantifying expression of LC3B RNA level using qPCR. A group of 20 healthy volunteers were also enrolled. RESULTS: Serum levels of PGRN and IL-6 as well as LC3B gene expression levels were markedly higher in type 2 diabetic patients. Additionally, our study revealed a cut-off value of 18.14 ng/mL for progranulin serum level and 3.23 for LC3B expression level, with sensitivities of 83.6% and 75.4% and specificities of 83.8% and 58.3%, respectively. Circulating PGRN levels are positively correlated with body mass index (BMI), glucose concentration, and IL-6. CONCLUSION: Our results support the hypothesis that progranulin is introduced as a novel marker of chronic inflammatory response in type 2 diabetes that aggravates insulin resistance via activated autophagy, indicating the importance of this novel adipokine in the regulation of glucose metabolism and as a promising therapeutic target in the treatment of diabetes. KEY WORDS: diabetes; progranulin; autophagy; microtubule-associated proteins light chain 3B; interleukin 6.

Caffeic Acid Attenuates Diabetic Kidney Disease via Modulation of Autophagy in a High-Fat Diet/Streptozotocin- Induced Diabetic Rat.

The aim of this study is to evaluate the anti-diabetic nephropathy effect of Caffeic acid and to prove our hypothesis for its mechanism of action that it may occur by reactivation of autophagy pathway via suppression of autophagy regulatory miRNAs. In vivo, high-fat diet and streptozotocin-induced (HFD-STZ) diabetic rats were treated with Caffeic acid once per day for 12 weeks before and after development of diabetic nephropathy. Blood and urine biochemical parameters, autophagy transcripts and their epigenetic regulators together with renal tissue morphology were investigated. In diabetic rats, Caffeic acid intake, caused improvement in albumin excretion,blood glucose, reduced renal mesangial matrix extension with increased vacuolation and reappearance of autophagosomes. Meanwhile, it resulted in autophagy genes up-regulation [RB 1-inducible coiled coil protein (RB1CC1), Microtubule-associated proteins 1A/1B light chain 3(MAP1LC3B), Autophagy related gene (ATG-12),] with simultaneous reduction in their epigenetic regulators; miRNA-133b, -342 and 30a, respectively. These above mentioned effects were more significant in the diabetic nephropathy Caffeic treated rats than in the prophylactic group. Based on our results we postulated that caffeic acid modulates autophagy pathway through inhibition of autophagy regulatory miRNAs, that could explain its curative properties against diabetic kidney disease.

Evaluation of urinary microRNA panel in bladder cancer diagnosis: relation to bilharziasis.

We assessed the differential expression of a urinary panel of microRNAs (miRs) in terms of potential application as diagnostic markers of bladder cancer (BC) and relationship to bilharziasis. We investigated voided urine samples and blood from patients with BC (n = 188), benign bladder lesions (n = 88), and age-matched controls (n = 92). Five miRs (miR-210, miR-10b, miR-29c, miR-221, and miR-23a) were selected from previous microarray signature profiling (released by miR2Disease). Afterward, they were validated using polymerase chain reaction array. The expression levels of miR-210, miR-10b, and miR-29c in the urine samples were significantly higher in BC (P < 0.001). The receiver-operating characteristic curve analyses demonstrated that each miR had good sensitivity and specificity for distinguishing patients with BC from patients without BC (miR-210, 71.3% and 91.1%; miR-10b, 80.9% and 91.1%; and miR-183, 71.3% and 88.9%). On combining the 3 miR detection data with the urinary cytology, the results sensitivity increased to 95.2%. Relative quantity mean rank of the miR-29c was significantly higher in the bilharzial-positive patients compared with bilharzial-negative patients. To conclude, urine miR-210, miR-10b, and miR-29c are promising tumor markers for BC: bilharzial and nonbilharzial.

Detection of 14q32 rearrangements in multiple myeloma, using simultaneous FISH analysis combined with immunofluorescence.

BACKGROUND: 14q32 rearrangement has been identified as a recurrent hotspot of translocations in multiple myeloma (MM). The Fluorescence Immunophenotyping and Interphase Cytogenetics as a tool for the Investigation of Neoplasms (known as FICTION technique) for evaluation of chromosomal changes in MM. The aim of this work is to detect 14q32 rearrangement, using FICTION technique, on archival bone marrow (BM) slides of MM patients, and to study its prognostic value. METHOD: This study was conducted at Ain Shams University Hospital. The FICTION technique, which uses CD138 and dual color, and the break-apart 14q32 rearrangement probe, was performed on archived smears of BM slides for 50 MM patients at the time of diagnosis. RESULTS: A significantly higher percentage of cases were positive for 14q32 rearrangement by FICTION (32%) compared to fluorescence in situ hybridization (FISH) (12%) (p=0.04). Cases positive by FICTION for the rearrangement were designated as Group A, while negative cases were designated as Group B. Significantly lower Hb and CRP levels were found among Group B when compared to Group A patients (p=0.001 and 0.01, respectively). Serum albumin level and Bence Jones protein (BJP) significantly affect overall survival (OS) (p=0.01, 0.007, respectively). However, a statistically non-significant shorter mean survival time was found in positive cases through FICTION versus negative cases. CONCLUSION: FICTION technique provides a sensitive tool for establishing clonal plasma cells (PC) infiltration of BM aspirates, and is amenable for use on archived as well as fresh smears.

Urinary retinoic acid receptor-ß2 gene promoter methylation and hyaluronidase activity as noninvasive tests for diagnosis of bladder cancer.

OBJECTIVES: We evaluated the significance of urinary retinoic acid receptor-ß2 (RAR-ß2) gene promoter methylation and hyaluronidase activity in comparison with voided urine cytology (VUC) in diagnosis of bladder cancer. DESIGN AND METHODS: This study included 100 patients diagnosed with bladder cancer, 65 patients with benign urological disorders and 51 healthy volunteers. Urine supernatant was used for determining hyaluronidase activity by zymography while urine sediment was used for cytology and detection of methylated RAR-ß2 gene promoter by methylation specific nested PCR. RESULTS: The sensitivity and specificity were 53% and 90.5% for VUC, 65% and 89.7% for percent methylation fraction of RAR-ß2 gene promoter, and 89% and 90.5% for hyaluronidase activity; combination of the three parameters increased sensitivity to 95%. A significant association was observed between investigated markers and advanced grade tumor. CONCLUSIONS: Combined use of RAR-ß2 gene promoter methylation, hyaluronidase activity and VUC is promising non-invasive tool for bladder cancer detection.

The diagnostic efficacy of urinary TGF-beta1 and VEGF in bladder cancer: comparison with voided urine cytology.

The purpose of this study was to evaluate the diagnostic efficacy of urinary transforming growth factor-beta1 (TGF-beta1) and vascular endothelial growth factor (VEGF) in comparison with voided urine cytology in the detection of bladder cancer. This study included 120 patients with bladder cancer, 54 patients with benign urological disorders and 55 healthy volunteers. Urine supernatant was used for estimation of TGF-beta1 and VEGF by ELISA. VEGF was detected by Western blot (WB) analysis in the urine supernatant of randomly selected bladder cancer patients. The urine sediment was used for cytology. There was a statistically significant difference in the median levels of TGF-beta1 (P=0.002) and VEGF (P=0.000) between the control, benign and malignant groups. The concordance rate of VEGF ELISA with VEGF WB was 96.3%. The overall sensitivity and specificity were 70.8% and 90.8% for voided urine cytology, 71.6% and 59.6% for TGF-beta1, and 76.7% and 61.5% for VEGF. The combined use of voided urine cytology with TGF-beta1 and VEGF improved the sensitivity up to 94.9%, although it lowered specificity to 62.0%. There was a significant association between positivity rate of TGF-beta1 and positive urine cytology samples (P=0.023). Median level and positivity rate of VEGF were significantly associated with early stage (I, II) of bladder carcinoma (P=0.01 and 0.025, respectively). Our data indicate that urinary TGF-beta1 and VEGF had higher sensitivities compared to voided urine cytology. Moreover, the combined sensitivity of voided urine cytology with TGF-beta1 and VEGF together was higher than sensitivity of voided urine cytology alone in detection of bladder cancer.