Humoral and cellular immune response to Buerger's disease.

OBJECTIVE: Thromboangiitis obliterans is a nonatherosclerotic occlusive disease, affecting small to moderate sized arteries of the upper and lower extremities, leading to progressive inflammation and clot formation. However, the role of humoral and cell-mediated immunity in the development of this disease has not been clearly identified. The present study was intended to investigate the humoral and cellular immune response in patients with Buerger's disease with different disease severity. METHODS: In an observational study, 80 male patients with Buerger's disease were included and categorized into three groups (mild, moderate, and severe) based on clinical manifestations. After blood sampling, cellular phenotypes were determined, and erythrocyte sedimentation rate, immunoglobulins (Ig) A, M, G, and E, as well as C3 and C4 components of the complement system and complement hemolytic activity (CH50) were measured. RESULTS: The mean age of the patient was 42.85 ± 8.39 years. Pulse abnormality, cold intolerance, and claudication were the most common symptoms. Eleven (13.75%), 46 (57.50%), and 23 (28.75%) patients had mild, moderate, and severe symptoms. Regression analyses showed that the presence of severe symptoms was significantly associated with elevated erythrocyte sedimentation rate and C4 levels (p < 0.05). CONCLUSION: Buerger's disease in severe cases was associated with increased erythrocyte sedimentation rate and abnormal C4 levels. The alterations in these inflammatory biomarkers might be due to a secondary inflammatory response to the presence of ulcer or gangrene and the inflammatory process in patients with severe symptoms.

Differentiation of adipocytes and osteocytes from human adipose and placental mesenchymal stem cells.

OBJECTIVES: Mesenchymal stem cells (MSC) can be isolated from adult tissues such as adipose tissue and other sources. Among these sources, adipose tissue (because of easy access) and placenta (due to its immunomodulatory properties, in addition to other useful properties), have attracted more attention in terms of research. The isolation and comparison of MSC from these two sources provides a proper source for clinical experimentation. The aim of this study was to compare the characteristics of MSC isolated from human adipose tissue and placenta. MATERIALS AND METHODS: Adipose and placental MSC were isolated from the subcutaneous adipose tissues of 10 healthy women (25 to 40 years) and from a fresh term placenta (n= 1), respectively. Stem cells were characterized and compared by flow cytometry using CD29, CD31, CD34, CD44, CD45, CD105, CD166 and HLA-DR markers. Osteocytes and adipocytes were differentiated from isolated human mesenchymal stem cells (HMSC). RESULTS: Adipose and placenta-derived MSC exhibited the same morphological features. ADSC differentiated faster than placenta; however, both were differentiated, taking up to 21 days for osteocyte and 14 days for adipocyte differentiation. About 90% of PLC-MSC and ADSC were positive for CD29, CD44, CD105, and CD166; and negative for CD31, CD34, CD45, and HLA-DR. CONCLUSION: The two sources of stem cells showed similar surface markers, morphology and differentiation potential and because of their multipotency for differentiating to adipocytes and osteocytes, they can be applied as attractive sources of MSC for regenerative medicine.

Evaluation of the cytotoxicity of fiber reinforced composite bonded retainers and flexible spiral wires retainers in simulated high and low cariogenic environments.

OBJECTIVES: The aim of this study was to evaluate the cytotoxic effects of fiber reinforced composite bonded retainers in comparison with flexible spiral wires (FSWs) under high and low cariogenic-simulated environments using human oral fibroblasts. MATERIALS AND METHODS: Four types of bonded retainers were evaluated: (1) reinforced with glass fibers: Interlig (Angelus), (2) reinforced with polyethylene fibers: Connect (Kerr), (3) reinforced with quartz fibers: Quartz Splint UD (RTD), and (4) FSW. Twenty specimens of each sample group were prepared with the same surface area and halved. Next, half of them were placed in a high cariogenic environment 60 min in 10% lactic acid 3 times a day and remained in Fusayama Meyer artificial saliva for the rest of the day) and the other half were placed in a low cariogenic environment 20 min in 10% lactic acid 3 times a day and remained in Fusayama Meyer artificial saliva for the rest of the day) for 1, 7 and 30 days. Cell viability was assessed by MTT assay. Data were analyzed using SPSS software (α =0.05). RESULTS: During the 1(st) month, cytotoxicity reduced gradually. In the low cariogenic-simulated environment, the cytotoxicity of all of the groups were reported to be mild at day 30 and the difference between them was significant (P = 0.016). In the same period in the high cariogenic-simulated environment, the cytotoxicity of Connect and Quartz Splint was mild, and they had lower cytotoxicity than the other groups. Meanwhile, Interlig had moderate (52%) and FSW had severe cytotoxicity (22%) and the difference between the groups was also significant (P = 0.000). CONCLUSIONS: FSW retainers are not recommended in those at high-risk for dental caries. However, in those at low-risk, there is no difference from the standpoint of cytotoxicity.

Antiproliferative activity and induction of apoptotic by ethanolic extract of Alpinia galanga rhizhome in human breast carcinoma cell line.

BACKGROUND: We investigated the potential of galangal rhizomes to induce cytotoxic and apoptotic effects in the cultured human breast carcinoma cell line, (MCF-7) in compare with the non-malignant (MRC-5) cells. METHODS: Both cells were cultured in DMEM medium and treated with galangal rhizomes for three consecutive days. The percentage of apoptotic cells was determined by flow cytometry using Annexin-V fluorescein isothiocyanate. RESULTS: The results showed that the ethanolic extract of galangal rhizomes decreased cell viability in the malignant cells as a concentration- and time- dependent manner. The IC50 values against MCF-7 were determined at 400.0 ± 11.7 and 170.0 ± 5.9 µg/ml after 48 and 72 h respectively. The morphology of MCF-7 cells treated with the ethanolic extract confirmed the cell proliferation assay results. Alpinia galanga induced apoptosis in MCF-7 cells, as determined by flow cytometry. CONCLUSIONS: We concluded that the extract of Alpinia galanga exerts pro-apoptotic effects in a breast cancer-derived cell line and could be considered as a potential chemotherapeutic agent in breast cancer.

Association between HLA-DQB1 alleles and HAM/TSP patients in Khorasan Province.

OBJECTIVES: HTLVI-1 is the first human retrovirus with limited endemic regions in the world. The epidemiological studies have shown that the genetic background and immune response to the virus have a significant role in HTLV-I-associated diseases. Among the genes are involved in HTLV-I infection, the role of human leukocytes antigen (HLA) have been studied in different population. In the present study we examined the association between HLA-DQB1 alleles and HTLV-I infection in HAM/TSP patients, HTLV-I carriers and healthy controls in north east of Iran, Mashhad. MATERIALS AND METHODS: The blood samples of 16 patients with HAM/TSP, 20 HTLV-1 carriers, and 30 healthy individuals were taken and DNA was extracted by salting out method. HLA-DQB1 typing was performed using PCR-SSP method and the frequency of HLA-DQB1 alleles were compared by Fischer Exact Test. RESULTS: There was a significant difference between HAM/TSP patients and healthy controls in the frequency of HLA-DQB1*07 (P=0.004, RR=7). Furthermore, we found that possession of HLA- DQB1*02 or HLA-DQB1*05 increased the risk of disease 1.5 times. CONCLUSION: The data presented here suggest that both HLA-DQB1*07 and HLA-DQB1*06 are associated with disease development.

Association between vitamin D receptor gene polymorphisms and type 1 diabetes mellitus in Iranian population.

Type 1 diabetes mellitus (T1DM) is one of the T-cell mediated autoimmune diseases and vitamin D suppresses activation of T-cell and has immunomodulatory effects. In this study the association between four vitamin D receptor (VDR) gene polymorphisms, at positions FokI, BsmI, ApaI and TaqI, and susceptibility to T1DM was investigated. We assessed 87 Iranian patients with T1DM and one hundred healthy controls with no history of diabetes or other autoimmune diseases. Our results demonstrated that genotypes frequency of the TaqI VDR polymorphism differed significantly between T1DM patients and controls, TT genotype and T allele was more frequent in healthy controls compared with TIDM patients (P = 0.003; OR = 0.51, 95% CI = 0.31-0.84). Therefore, allele t is the risk-allele for developing TIDM in this study. No significant association was observed between others VDR SNPs and disease susceptibility. In conclusion, our case-control study indicated that the VDR TaqI polymorphism is associated with TIDM in Iranian population.

Chrysin reduces proliferation and induces apoptosis in the human prostate cancer cell line pc-3.

INTRODUCTION: Honey is a common household product with many medicinal uses described in traditional medicine. Only recently has its antioxidant properties and preventive effects against disease been highlighted. Chrysin is a natural flavone commonly found in honey that has been shown to be an antioxidant agent. In this study, we investigated the antiproliferative and apoptotic effects of honey and chrysin on cultured human prostate cancer cells. METHODS: Cells were cultured in RPMI medium and treated with different concentrations of honey and chrysin for three consecutive days. Cell viability was quantitated by the 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. The percentage of apoptotic cells was determined by flow cytometry using Annexin V-fluorescein isothiocyanate. RESULTS: The MTT assay revealed that both compounds had an antiproliferative effect on PC-3 cells in a dose- and time-dependent manner. The IC50 values for honey and chrysin against PC-3 cells were 2.5% and 24.5% after 48 h and 1.8% and 8.5% after 72 h, respectively. Chrysin induced apoptosis in PC-3 cells, as determined by flow cytometry. CONCLUSION: Our results suggest that honey has anti-proliferative effects on prostate cancer cells and the effects are mainly due to chrysin. Therefore, chrysin may be a potential compound for both cancer prevention and treatment. Further in vivo investigation is needed to support the use of chrysin in cancer therapy.

Honey induces apoptosis in renal cell carcinoma.

BACKGROUND: The fact that antioxidants have several preventative effects against different diseases, such as coronary diseases, inflammatory disorders, neurologic degeneration, aging, and cancer, has led to the search for food rich in antioxidants. Honey has been used as a traditional food and medical source since ancient times. However, recently many scientists have been concentrating on the antioxidant property of honey. By use of human renal cancer cell lines (ACHN), we investigated the antiproliferative activity, apoptosis, and the antitumor activity of honey. MATERIALS AND METHODS: The cells were cultured in Dulbecco's modified Eagle's medium with 10% fetal bovine serum treated with different concentrations of honey for 3 consecutive days. Cell viability was quantitated by the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptotic cells were determined using Annexin-V-fluorescein isothiocyanate (FITC) by flow cytometry. RESULTS: Honey decreased the cell viability in the malignant cells in a concentration- and time-dependent manner. The IC (50) values against the ACHN cell lines were determined as 1.7 ± 0.04% and 2.1 ± 0.03% µg/mL after 48 and 72 h, respectively. Honey induced apoptosis of the ACHN cells in a concentration-dependent manner, as determined by flow cytometry histogram of treated cells. CONCLUSION: It might be concluded that honey may cause cell death in the ACHN cells, in which apoptosis plays an important role. Most of the drugs used in the cancer treatment are apoptotic inducers, hence apoptotic nature of honey is considered vital. Therefore, it prompted us to investigate honey as a potential candidate for renal cancer treatment.

Chrysin reduces proliferation and induces apoptosis in the human prostate cancer cell line pc-3

INTRODUCTION: Honey is a common household product with many medicinal uses described in traditional medicine. Only recently has its antioxidant properties and preventive effects against disease been highlighted. Chrysin is a natural flavone commonly found in honey that has been shown to be an antioxidant agent. In this study, we investigated the antiproliferative and apoptotic effects of honey and chrysin on cultured human prostate cancer cells. METHODS: Cells were cultured in RPMI medium and treated with different concentrations of honey and chrysin for three consecutive days. Cell viability was quantitated by the 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. The percentage of apoptotic cells was determined by flow cytometry using Annexin V-fluorescein isothiocyanate. RESULTS: The MTT assay revealed that both compounds had an antiproliferative effect on PC-3 cells in a dose- and time-dependent manner. The IC50 values for honey and chrysin against PC-3 cells were 2.5 percent and 24.5 percent after 48 h and 1.8 percent and 8.5 percent after 72 h, respectively. Chrysin induced apoptosis in PC-3 cells, as determined by flow cytometry. CONCLUSION: Our results suggest that honey has anti-proliferative effects on prostate cancer cells and the effects are mainly due to chrysin. Therefore, chrysin may be a potential compound for both cancer prevention and treatment. Further in vivo investigation is needed to support the use of chrysin in cancer therapy.

Cytotoxic effect of saffron stigma aqueous extract on human transitional cell carcinoma and mouse fibroblast.

INTRODUCTION: Saffron has been suggested to have inhibitory effects on tumoral cells. We evaluated the cytotoxic effect of aqueous extract of saffron on human transitional cell carcinoma (TCC) and mouse non-neoplastic fibroblast cell lines. MATERIALS AND METHODS: Human TCC 5637 cell line and mouse fibroblast cell line (L929) were cultivated and incubated with different concentrations of aqueous extract of saffron stigma (50 microg/mL to 4000 microg/mL). Cytotoxic effect of saffron was evaluated by morphologic observation and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay after 24, 48, 72, and 120 hours in each cell line. RESULTS: After 24 hours, morphological observations showed growth inhibitory effects at saffron extract concentrations higher than 200 microg/mL for L929 cells and at concentrations of 50 microg/mL to 200 microg/mL for the TCC cells. These changes became more prominent after 48 hours. However, significant growth inhibitory effects of the extract were shown at concentrations of 400 microg/mL and 800 microg/mL. Higher concentrations of saffron correlated inversely with cell population of both cell lines. Significant reduction of the survived cells was seen at concentrations of 400 microg/mL and 2000 microg/mL for TCC and L929 cell lines, respectively. After 120 hours, decrease in the percentage of survived cells at higher concentrations of saffron extract was seen in both cell lines. At a concentration of 800 microg/mL, the survived L929 cells plummeted to less than 60% after 120 hours, while no TCC cells survived at this time. No L929 cells survived at 2000 microg/mL. CONCLUSION: Saffron aqueous extract has inhibitory effects on the growth of both TCC 5637 and normal L929 cell lines. This effect is dose dependent.

Increased microvessel density in involved organs from patients with HTLV-I associated adult T cell leukemia lymphoma.

Adult T-cell leukemia-lymphoma (ATLL) is a rapidly progressive lymphoproliferative disorder secondary to infection with the human T cell lymphotropic virus type I (HTLV-I). The role of angiogenesis in the development and prognosis of many hematologic malignancies is established. We have previously shown that ATLL derived cells secrete high levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b-FGF), induce endothelial tube formation in vitro and establish functional gap junction-mediated communication with endothelial cells. We also demonstrated that plasma from ATLL and tropical spastic paraparesis/HTLV-I associated myelopathy patients exhibit very high levels of VEGF and b-FGF. Recently, we showed that treatment with the combination of zidovudine and interferon alpha reduced both HTLV-I proviral load and importantly VEGF plasma levels suggesting a potential anti-angiogenic effect of this therapy. In this report, we evaluated microvessel density (MVD) in involved organs from 20 patients with ATLL, as compared to normal organs from matched controls. We show evidence of significantly increased MVD in all tested involved organs from ATLL patients, suggesting that angiogenesis plays an important role in the development or organ invasion of ATLL, and could represent a potentially interesting target for anti-angiogenic therapy of ATLL.