Differential cytotoxic effects of fenbendazole on mouse lymphoma EL-4 cells and spleen cells

Fenbendazole (FBZ) is a benzimidazole anthelmintic widely used to treat parasitic infections. The anticancer effect of FBZ has been recently highlighted leading to its consideration as a potential anticancer agent. Although previous studies have demonstrated the effect of FBZ on cancer cells, there is a paucity of studies on the effect of FBZ on lymphoma cells and normal immune cells. Herein, we investigated the effects of FBZ on a mouse lymphoma cell line, EL-4 cells, and spleen cells, using vincristine as a positive control. The cellular metabolic activity of EL-4 cells was decreased by FBZ, but that of the spleen cells was not decreased. Moreover, FBZ reduced the mitochondrial membrane potential and induced reactive oxygen species production in EL-4 cells, but not in spleen cells. FBZ induced G2/M phase arrest and increased the sub G0/G1 phase ratio, indicating apoptosis. Furthermore, compared to the control cells, the reactivity of spleen cells pretreated with FBZ to lipopolysaccharide was maintained. In summary, FBZ is cytotoxic to EL-4 cells, but not to spleen cells. This study provides experimental evidence that FBZ exerts an anticancer effect, and less cytotoxic effects and functional damage to normal spleen cells.

Biphasic immunomodulatory effects of ionized biosilica water on the antigen-presenting capability of mouse dendritic cells

Biosilica is a silica-based substance derived from the cell walls (frustules) of diatoms and has various biological activities. Recently, research into biosilica’s biological functions are underway, but little has been reported on the effects of biosilica on immune cells. In this study, we investigated the effect of ionized biosilica water (iBW) on dendritic cells (DCs), which play crucial roles as antigen (Ag)-presenting cells. Treatment with iBW increased the expression of immune response-related markers, closely connected to the maturation of DCs, and the production of tumor necrosis factor-alpha. In addition, iBW-treated DCs (iBW-DCs) had a lower uptake of fluorescein isothiocyanate-dextran than that of control DCs. Mixed leukocyte response analysis, used for measuring the Ag-presenting capability of DCs, showed iBW-DCs had a higher capability than that of control DCs. Interestingly, DCs treated with lipopolysaccharide (LPS) and iBW had a lower level of Ag-presenting capability than that of LPS-treated DCs. Taken together, the results indicate that iBW alone can mature DCs, but it decreases the Ag-presenting capability of DCs in the presence of LPS, a representative agent of inflammation. This study may provide valuable information regarding the effect of iBW on immune cells. Further research is needed to investigate how iBW induces the observed biphasic immunomodulatory activity.

Biphasic immunomodulatory effects of ionized biosilica water on the antigen-presenting capability of mouse dendritic cells

Biosilica is a silica-based substance derived from the cell walls (frustules) of diatoms and has various biological activities. Recently, research into biosilica’s biological functions are underway, but little has been reported on the effects of biosilica on immune cells. In this study, we investigated the effect of ionized biosilica water (iBW) on dendritic cells (DCs), which play crucial roles as antigen (Ag)-presenting cells. Treatment with iBW increased the expression of immune response-related markers, closely connected to the maturation of DCs, and the production of tumor necrosis factor-alpha. In addition, iBW-treated DCs (iBW-DCs) had a lower uptake of fluorescein isothiocyanate-dextran than that of control DCs. Mixed leukocyte response analysis, used for measuring the Ag-presenting capability of DCs, showed iBW-DCs had a higher capability than that of control DCs. Interestingly, DCs treated with lipopolysaccharide (LPS) and iBW had a lower level of Ag-presenting capability than that of LPS-treated DCs. Taken together, the results indicate that iBW alone can mature DCs, but it decreases the Ag-presenting capability of DCs in the presence of LPS, a representative agent of inflammation. This study may provide valuable information regarding the effect of iBW on immune cells. Further research is needed to investigate how iBW induces the observed biphasic immunomodulatory activity.

Bordetella bronchiseptica is a potent and safe adjuvant that enhances the antigen-presenting capability of dendritic cells

We previously demonstrated that Bordetella bronchiseptica (B. bronchiseptica) antigen (Ag) enhances the Mycoplasma hyopneumoniae Ag-specific immune response. The focus of this study was whether acellular bacterin of B. bronchiseptica could be used as an adjuvant to increase antigen-presenting capability of dendritic cells (DCs) by increasing the level of activation. The metabolic activity of DCs was increased by B. bronchiseptica, similar to lipopolysaccharide (LPS). Flow cytometry analysis revealed that B. bronchiseptica increases the expression of major histocompatibility complex class-2, cluster of differentiation (CD)40, CD54, and CD86 which are closely related to DC-mediated immune responses. B. bronchiseptica enhanced the production of cytokines related to adaptive immune responses. Furthermore, the survival rate of B. bronchiseptica-injected groups was 100% at 15 and 20 mg/kg doses, whereas that of LPS-injected groups was only 20%, 0% at 15 and 20 mg/kg doses respectively, and so B. bronchiseptica is likely to be safer than LPS. Taken together, these results indicate that B. bronchiseptica can be used as an adjuvant to enhance the antigen-presenting capability of DCs. B. bronchiseptica is a candidate for producing vaccines, especially in case of DC-mediating efficacy and safety demands. This study provides researchers and clinicians with valuable information regarding the usage of B. bronchiseptica as a safe bacteria-derived immunostimulating agent for developing efficient vaccines.

Lineage Differentiation Program of Invariant Natural Killer T Cells

Invariant natural killer T (iNKT) cells are innate T cells restricted by CD1d molecules. They are positively selected in the thymic cortex and migrate to the medullary area, in which they differentiate into 3 different lineages. Promyelocytic leukemia zinc finger (PLZF) modulates this process, and PLZFhigh, PLZFintermediate, and PLZFlow iNKT cells are designated as NKT2, NKT17, and NKT1 cells, respectively. Analogous to conventional helper CD4 T cells, each subset expresses distinct combinations of transcription factors and produces different cytokines. In lymphoid organs, iNKT subsets have unique localizations, which determine their cytokine responses upon antigenic challenge. The lineage differentiation programs of iNKT cells are differentially regulated in various mice strains in a cell-intrinsic manner, and BALB/c mice contain a high frequency of NKT2 cells. In the thymic medulla, steady state IL-4 from NKT2 cells directly conditions CD8 T cells to become memory-like cells expressing Eomesodermin, which function as premade memory effectors. The genetic signature of iNKT cells is more similar to that of γδ T cells and innate lymphoid cells (ILCs) than of conventional helper T cells, suggesting that ILCs and innate T cells share common developmental programs.

Analyses of the TCR repertoire of MHC class II-restricted innate CD4+ T cells

Analysis of the T-cell receptor (TCR) repertoire of innate CD4+ T cells selected by major histocompatibility complex (MHC) class II-dependent thymocyte-thymocyte (T-T) interaction (T-T CD4+ T cells) is essential for predicting the characteristics of the antigens that bind to these T cells and for distinguishing T-T CD4+ T cells from other types of innate T cells. Using the TCRmini Tg mouse model, we show that the repertoire of TCRalpha chains in T-T CD4+ T cells was extremely diverse, in contrast to the repertoires previously described for other types of innate T cells. The TCRalpha chain sequences significantly overlapped between T-T CD4+ T cells and conventional CD4+ T cells in the thymus and spleen. However, the diversity of the TCRalpha repertoire of T-T CD4+ T cells seemed to be restricted compared with that of conventional CD4+ T cells. Interestingly, the frequency of the parental OT-II TCRalpha chains was significantly reduced in the process of T-T interaction. This diverse and shifted repertoire in T-T CD4+ T cells has biological relevance in terms of defense against diverse pathogens and a possible regulatory role during peripheral T-T interaction.

Subclinical hypothyroidism diagnosed by thyrotropin-releasing hormone stimulation test in infertile women with basal thyroid-stimulating hormone levels of 2.5 to 5.0 mIU/L

OBJECTIVE: To investigate the prevalence of subclinical hypothyroidism (SH) diagnosed by thyrotropin-releasing hormone (TRH) stimulating test in infertile women with basal thyroid-stimulating hormone (TSH) levels of 2.5 to 5.0 mIU/L. METHODS: This study was performed in 39 infertile women with ovulatory disorders (group 1) and 27 infertile women with male infertility only (group 2, controls) who had basal serum TSH levels of 2.5 to 5.0 mIU/L and a TRH stimulating test. Serum TSH levels were measured before TRH injection (TSH0) and also measured at 20 minutes (TSH1) and 40 minutes (TSH2) following intravenous injection of 400 microg TRH. Exaggerated TSH response above 30 mIU/L following TRH injection was diagnosed as SH. Group 1 was composed of poor responders (subgroup A), patients with polycystic ovary syndrome (subgroup B) and patients with WHO group II anovulation except poor responder or polycystic ovary syndrome (subgroup C). RESULTS: The prevalence of SH was significantly higher in group 1 of 46.2% (18/39) compared with 7.4% (2/27) in group 2 (P=0.001). TSH0, TSH1, and TSH2 levels were significantly higher in group 1 than the corresponding values in group 2 (P<0.001, P<0.001, P<0.001). In group 1, TSH1 and TSH2 levels were significantly lower in subgroup C compared with those in subgroup A and B (P=0.008, P=0.006, respectively). CONCLUSION: TRH stimulation test had better be performed in infertile women with ovulatory disorders who have TSH levels between 2.5 and 5.0 mIU/L for early detection and appropriate treatment of SH.

The effect of aromatase inhibitor letrozole incorporated in gonadotrophin-releasing hormone antagonist multiple dose protocol in poor responders undergoing in vitro fertilization

OBJECTIVE: To evaluate whether letrozole incorporated in a gonadotrophin-releasing hormone (GnRH) antagonist multiple dose protocol (MDP) improved controlled ovarian stimulation (COS) and in vitro fertilization (IVF) results in poor responders who underwent IVF treatment. METHODS: In this retrospective cohort study, a total of 103 consecutive IVF cycles that were performed during either the letrozole/GnRH antagonist MDP cycles (letrozole group, n=46) or the standard GnRH antagonist MDP cycles (control group, n=57) were included in 103 poor responders. COS results and IVF outcomes were compared between the two groups. RESULTS: Total dose and days of recombinant human follicle stimulating hormone (rhFSH) administered were significantly fewer in the letrozole group than in the control group. Duration of GnRH antagonist administered was also shorter in the letrozole group. The number of oocytes retrieved was significantly higher in the letrozole group. However, clinical pregnancy rate per cycle initiated, clinical pregnancy rate per embryo transfer, embryo implantation rate and miscarriage rate were similar in the two groups. CONCLUSION: The letrozole incorporated in GnRH antagonist MDP may be more effective because it results comparable pregnancy outcomes with shorter duration and smaller dose of rhFSH, when compared with the standard GnRH antagonist MDP.

The effect of luteal phase progesterone supplementation on natural frozen-thawed embryo transfer cycles

OBJECTIVE: To evaluate the effect of progesterone supplementation during the luteal phase on pregnancy outcome in natural frozen-thawed embyo transfer (FTET) cycles. METHODS: In this retrospective cohort study, 228 consecutive patients who underwent FTET cycles between January 2009 and September 2012 were included. One hundred forty-five patients received luteal progesterone support (P group) but 83 patients did not receive any progesterone supplementation during luteal phase (control group). RESULTS: There were no differences in patients' characteristics between the two groups. The two groups were similar with respect to the characteristics of previous fresh in vitro fertilization cycle in which embryos were cryopreserved including the numbers of oocytes retrieved, mature oocytes, fertilized oocytes, grade 1 or 2 embryos and frozen embryos. Also, significant differences were not observed between the P and control groups in clinical pregnancy rate, embryo implantation rate and multiple pregnancy rate. However, miscarriage rate was significantly lower in the P group and live birth rate was significantly higher in the P group than in the control group (P<0.05, P<0.05). CONCLUSION: Our results suggest that luteal phase progesterone supplementation decreases miscarriage rate and improves live birth rate in natural FTET cycles.

Surgical resection or aspiration with ethanol sclerotherapy of endometrioma before in vitro fertilization in infertilie women with endometrioma

OBJECTIVE: To evaluate whether the surgical resection or aspiration with ethanol sclerotherapy (AEST) of endometrioma before in vitro fertilization (IVF) affect controlled ovarian stimulation (COS) and IVF outcome in the infertilie women with endometroma undergoing IVF. METHODS: In this retrospective cohort study, 101 consecutive IVF/intracytoplasmic sperm injection cycles that were performed in 101 patients with endometrioma(s) between January 2008 and December 2012 were included. Before IVF, 36 patients underwent surgical resection of endometrioma (resection group), 29 patients had transvaginal endometrioma AEST (aspiration group), and 36 patients did not take any surgical intervention (control group). The three groups were compared in terms of COS and IVF outcomes. RESULTS: Total antral follicle count was significantly lower in the resection group than in the aspiration or control group. The numbers of follicles with a diameter of 14 to 17 mm on the human chorionic gonadotropin day, retrieved oocytes, mature oocytes, and fertilized oocytes were significantly lower in the resection group than in two other groups. However, three groups were similar in terms of clinical pregnancy rate (CPR) per initiated cycle, CPR per embryo transfer, embryo implantation rate, and miscarriage rate. CONCLUSION: Neither of surgical resection and AEST of endometrioma before IVF treatment can give any beneficial effect on IVF outcomes. Moreover, surgical resection of endometrioma can affect the ovarian reserve and ovarian response during COS.

Undifferentiated Carcinoma Arising in a Choledochal Cyst: A Case Report

An association between choledochal cyst and carcinoma is well established. Here, we report an extremely rare case of undifferentiated carcinoma exhibiting extensive sarcomatous features arising in a choledochal cyst. The patient in our case had a radiologically confirmed choledochal cyst and anomalous pancreaticobiliary ductal union, and mild wall thickening in the cyst was observed on endoscopic retrograde cholangiopancreatography. The patient underwent common bile duct excision and cholecystectomy. In the choledochal cyst, a nodule measuring 1.5x1 cm was detected. The lesion was composed of atypical, spindle-shaped and large, round pleomorphic tumor cells simulating sarcoma. Neither glandular nor squamous differentiation was observed. These cells were immunoreactive for both vimentin and cytokeratin by immunohistochemistry. These histologic and immunohistochemical findings were consistent with undifferentiated carcinoma, spindle and giant cell type, according to the WHO classification.

Pathologic Features of Korean Prostate Adenocarcinoma: Mapping Analysis of 83 Cases

BACKGROUND :Prostatic adenocarcinoma makes up about 2% of the total cancer incidence and cancer death in Korean men, but the incidence of this malady is continuously increasing. So far, there have been only a few studies describing the pathologic characteristics of the prostatic adenocarcinoma in Korean patients. In this study, we analyzed 83 radical prostatectomy specimens by using mapping analysis to discover the clinico pathologic characteristics of Korean prostatic adenocarcinoma. METHODS: The resected prostates were serially sectioned and embedded for histologic mapping. The clinico pathologic findings, including the Gleason score, tumor size, prostate intraepithelial neoplasia (PIN) and tumor invasion to the surrounding tissues, were examined. RESULTS: The mean values were as follows: age, 64.1+/-6.6 years; serum prostate specific antigen (sPSA), 16.6+/-16.2 ng/mL; tumor volume, 22.3+/-22.4%; tumor size, 2.2+/-1.2 cm; and Gleason score, 6.9+/-0.9. The rate of high grade PIN was 79.7%. The Gleason score, tumor extent and T stage were statistically correlated (p<0.05). CONCLUSIONS: Some prognostic factors such as sPSA and the Gleason scores showed significantly lower levels compared with those of the previous studies on Korean prostate adenocarcinoma (16-36 ng/mL vs 16.6 ng/mL and 7.3-7.7 vs 6.9, respectively). Although these values are still higher than those of the western studies, this study implies that the early detection of prostate adenocarcinoma is increasing in Korea.

Epithelial Cysts in the Intrapancreatic Accessory Spleen that Clinically Mimic Pancreatic Cystic Tumor: A Report of Two Cases

Cystic lesions in the accessory spleen are extremely rare and they present a challenging clinical differential diagnosis. We report here on two cases of epithelial cyst of intrapancreatic accessory spleen that mimicked pancreatic cystic tumor. In both cases, the patients underwent distal pancreatectomy under the impression of a benign cystic tumor of the pancreas. Unilocular or multilocular cysts in the pancreas tail were observed, and these were later shown to be epithelial cysts in the accessory spleen located within the pancreatic tail. The cysts were lined by columnar, cuboidal or stratified squamous epithelium.