Retraction Note: Roles of the Nrf2/HO-1 pathway in the anti-oxidative stress response to ischemia-reperfusion brain injury in rats.

The article "Roles of the Nrf2/HO-1 pathway in the anti-oxidative stress response to ischemia-reperfusion brain injury in rats", by L.-J. Jiang, S.-M. Zhang, C.-W. Li, J.-Y. Tang, F.-Y. Che, Y.-C. Lu, published in Eur Rev Med Pharmacol Sci 2017; 21 (7): 1532-1540-PMID: 28429353 has been retracted by the Editor in Chief. Following some concerns raised on PubPeer (link: https://pubpeer.com/publications/4C502B6EB4FCA59AC9F42A8278A3D4), the Editor in Chief has started an investigation to assess the validity of the results as well as possible figure manipulation. The authors have been informed about the journal's investigation but remained unresponsive and have not provided the study's raw data. The journal investigation revealed several figure duplications and manipulations in Figures 3 and 6. Consequently, the Editor in Chief mistrusts the results presented and has decided to retract the article. This article has been retracted. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/12521.

The impact of transferred air bubble position on clinical pregnancy rate in FET cycles.

We aim to investigate the correlation of the position of the transferred air bubble with the clinical pregnancy rate (PR) in frozen-thawed embryo transfer(FET) cycles. A prospective clinical study was carried out at Reproductive medicine center of West China Second University Hospital between June 2020 and May 2021. 1159 women underwent FET were included in this study. Transabdominal ultrasonographic guidance was used during the transfer procedure. The distance from the air bubble to endometrial cavity fundus（DAF）was measured in the freeze-frame ultrasound immediately after ET. In group DAF ≤3mm, 3-15mm and ≥15mm, the clinical PR in women transferred with cleavage embryos were 33.3% (7/21), 55.0% (153/280), and 31.3% (5/16), respectively, the difference was statistically significant (P＜0.05). Among women transferred with blastocysts, the clinical PR was 63.0% (34/54), 68.5% (485/708) and 55.0% (44/80), respectively, the difference was statistically significant (P＜0.05). In multivariate logistic regression model for clinical PR, the clinical PR was associated with age, embryo quality, number of embryo transferred, and endometrial thickness. DAF was an independent risk factor influencing clinical PR in blastocysts FET cycles rather than in cleavage embryos FET cycles.In conclusion, our results suggested that DAF was associated with the clinical PR and DAF between 3mm and 15mm is the optimal position in blastocysts FET cycles.

Inter-relationship of risk factors and pathways associated with chronic kidney disease in patients with type 2 diabetes mellitus: a structural equation modelling analysis.

OBJECTIVES: Diabetes mellitus is the most common cause of chronic kidney disease (CKD); however, the inter-relationships and pathogenetic mechanisms among risk factors are still largely unknown. Structural equation modelling (SEM) was applied to test a hypothesis of causal pathways related to CKD in patients with type 2 diabetes mellitus (T2DM). STUDY DESIGN: This is a prospective observational study. METHODS: A total of 3395 patients with T2DM were enrolled in this study. A hypothesised SEM was applied to assess associations among demographic data, diabetic self-management behaviours, diabetes control, lifestyle, psycho-social, chronic inflammation factors, anthropometric and metabolic variables simultaneously and the risk of CKD. RESULTS: Demographic data (including education, marital status and mini-mental state examination score) (-0.075), white blood cell count (0.084), high blood pressure (0.144), World Health Organisation (WHO) 5 well-being index (-0.082), diabetes control (0.099), triglyceride (0.091) and uric acid (0.282) levels had direct effects on the risk of CKD. The final model could explain 26% of the variability in baseline CKD status. In addition, the same direct and specific indirect factors at baseline CKD status analysis contributed to the risk of CKD at the 12-month follow-up. The final model could explain 31% of the variability in the risk of CKD at the 12-month follow-up. CONCLUSIONS: This study investigates associations between factors obtained from real-world daily practice and CKD status simultaneously and delineates the potential pathways and inter-relationships of the risk factors that contribute to the development of CKD in patients with T2DM.

Hyper-viscoelastic mechanical behavior of cranial pia mater in tension.

BACKGROUND: Cranial pia mater, the innermost layer of the meninges, protects the central nervous system by tightly wrapping the brain and damping the external impact force to the brain. Accurate experimental data of the mechanical property of the cranial pia mater can enhance the theoretical prediction of traumatic brain injury or the scientific surgery design for brain disease. The aim of this study is to characterize the mechanical behavior of the cranial pia mater. METHODS: In vitro tensile and stress-relaxation experiments of ovine cranial pia mater specimens were conducted at eight strain rates to characterize the rate-dependent viscoelastic property. The tensile and stress-relaxation experimental data were fitted by an Ogden hyper-viscoelastic model with a strain rate function to describe the mechanical behavior of the cranial pia mater. FINDINGS: The elastic modulus and the ultimate stress are significantly increased from 5.545 MPa and 0.535 MPa at 0.00167 s-1 to 18.345 MPa and 2.547 MPa at 0.83 s-1 (p < .0001), respectively. The initial stress and the long-term stress (300 s) are also increased significantly with the increasing strain rates (p < .0001). A good fit of the experimental data with the Ogden hyper-viscoelastic model incorporated with a strain rate function was achieved (R2 > 0.93). INTERPRETATION: The cranial pia mater exhibits as a rate-dependent hyper-viscoelastic material in the tensile and stress-relaxation experiments. Compared with the brain, the stiffer nature of the cranial pia mater indicates its essential role in brain protection. The rate-dependent constitutive model provides a proper description of the hyper-viscoelastic characteristics of the cranial pia mater in tension and may provide a basic constitutive relationship for numerical simulations of traumatic brain injury.

Action of the insecticide cyfluthrin on Ca2+ signal transduction and cytotoxicity in human osteosarcoma cells.

Cyfluthrin is a pyrethroid insecticide and common household pesticide. The effect of cyfluthrin on Ca2+-related physiology in human osteosarcoma is unclear. This study investigated the effect of cyfluthrin on cytosolic-free Ca2+ concentrations ([Ca2+]i) and viability in MG63 human osteosarcoma cells. Cyfluthrin concentration-dependently induced [Ca2+]i rises. Cyfluthrin-induced Ca2+ entry was confirmed by the Mn2+-induced quench of fura-2 fluorescence. Cyfluthrin at concentrations of 10-100 µM induced [Ca2+]i rises. Ca2+ removal reduced the signal by approximately 50%. Cyfluthrin (100 µM) induced Mn2+ influx suggesting Ca2+ entry. Cyfluthrin-induced Ca2+ entry was inhibited 50% by protein kinase C (PKC) activator (phorbol 12-myristate 13-acetate) and inhibitor (GF109203X) and also by three inhibitors of store-operated Ca2+ channels: nifedipine, econazole, and SKF96365. In Ca2+-free medium, treatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin (TG) completely inhibited cyfluthrin-evoked [Ca2+]i rises. Conversely, treatment with cyfluthrin abolished TG-evoked [Ca2+]i rises. Inhibition of phospholipase C (PLC) with 1-[6-[((17ß)-3-methoxyestra-1,3,5[10]-trien-17-yl)amino]hexyl]-1H-pyrrole-2,5-dion abolished cyfluthrin-induced [Ca2+]i rises. Cyfluthrin at 25-65 µM decreased cell viability, which was not reversed by pretreatment with the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester. Together, in MG63 cells, cyfluthrin induced [Ca2+]i rises by evoking PLC-dependent Ca2+ release from the endoplasmic reticulum and Ca2+ entry via PKC-sensitive store-operated Ca2+ entry. Cyfluthrin also caused Ca2+-independent cell death.

A 3D Computational Head Model Under Dynamic Head Rotation and Head Extension Validated Using Live Human Brain Data, Including the Falx and the Tentorium.

We employ an advanced 3D computational model of the head with high anatomical fidelity, together with measured tissue properties, to assess the consequences of dynamic loading to the head in two distinct modes: head rotation and head extension. We use a subject-specific computational head model, using the material point method, built from T1 magnetic resonance images, and considering the anisotropic properties of the white matter which can predict strains in the brain under large rotational accelerations. The material model now includes the shear anisotropy of the white matter. We validate the model under head rotation and head extension motions using live human data, and advance a prior version of the model to include biofidelic falx and tentorium. We then examine the consequences of incorporating the falx and tentorium in terms of the predictions from the computational head model.

Effects of female sex hormones on folic acid-induced anti-angiogenesis.

AIM: Pregnant women have been recommended to take FA daily to prevent birth defects in the brain and spinal cord. We previously showed that folic acid (FA) exerts an anti-angiogenic activity. As angiogenesis is important for endometrial reorganization and embryonic development, there should be some mechanisms to allow the pregnant mother and the foetus to escape from the FA-induced anti-angiogenesis. This study was designed to investigate the effect of female sex hormones on the FA-induced anti-angiogenic activity. METHODS: The protein levels and protein-protein interaction were examined by Western blot analysis and immunoprecipitation assay respectively. The cell proliferation and migration were examined by MTT assay and wound healing assay respectively. The in vivo angiogenesis was evaluated by Matrigel angiogenesis assay. RESULTS: In human umbilical venous endothelial cells (HUVEC), FA receptor (FR) formed a complex with progesterone receptor (PR), oestradiol receptor (ER) and cSrc. Pregnancy levels of progesterone (P4) or oestradiol (E2) prevented FA-induced inhibitions of proliferation and migration in HUVEC. Both E2 and P4 prevented the FA-induced anti-angiogenesis in vivo. Moreover, cotreatment with FA and P4 or E2 inhibited the signalling pathways involved in FA-induced inhibitions of proliferation and migration in HUVEC. CONCLUSION: Female sex hormones interrupt the FA-induced anti-angiogenic action through receptor-receptor interaction.

Inhibition of cyclooxygenase-2-mediated matriptase activation contributes to the suppression of prostate cancer cell motility and metastasis.

Chronic inflammation plays an important role in cancer development and progression. Cyclooxygenases-2 (COX-2) is a key enzyme in generating prostaglandins causing inflammation, is often found to be overexpressed in prostate cancer (PCa) and is correlated with PCa cell invasion and metastasis. We aim to investigate the molecular mechanism of how COX-2 promotes PCa cell invasion and metastasis and to evaluate the effect of COX-2 inhibitors in a selected model of PCa progression. Our results showed that the expression of COX-2 and Interleukin 1ß (IL-1ß) was upregulated in highly invasive PCa cells and was correlated with the activated levels of membrane-anchored serine protease matriptase. The expression levels of COX-2 were increased and were correlated with matriptase levels in PCa specimens. Moreover, results showed that COX-2 overexpression or a COX-2 product Prostaglandin E2 (PGE2) caused an increase in matriptase activation and PCa cell invasion, whereas COX-2 silencing antagonized matriptase activation and cell invasion. In addition, the inhibition of COX-2-mediated matriptase activation by Celebrex and sulindac sulfide suppressed the androgen-independent and COX2-overexpressing PCa PC-3 cell invasion, tumor growth and lung metastasis in an orthotopic xenograft model. Our results indicate that COX-2/matriptase signaling contributes to the invasion, tumor growth and metastasis of COX-2-overexpressing and androgen-independent PCa cells.

Effects of Gender and Age on Immune Responses of Human Peripheral Blood Mononuclear Cells to Probiotics: A Large Scale Pilot Study.

BACKGROUND: Despite the widely accepted concept that probiotics confer miscellaneous benefits to hosts, the controversies surrounding these health-promoting claims cannot be ignored. These controversies hinder development and innovation in this field. RESULTS: To clarify the effects of age and gender on probiotic-induced immune responses, we recruited 1613 Taiwanese individuals and calculated the ratio of IFN-γ to IL-10 production after each individual's PBMCs were stimulated by six probiotic strains (L. paracasei BRAP01, L. acidophilus AD300, B. longum BA100, E. faecium BR0085, L. rhamnosus AD500 and L. reuteri BR101). Our results indicated that gender and age have only minor effects on the immune modulation of probiotics. Additionally, we showed that L. paracasei BRAP01 and L. acidophilus AD300 are the two dominant strains inducing IFN-γ/IL-10 production in Taiwanese individuals and that L. reuteri BR101 was the most effective stimulator of IL-10/IFN-γ. Additionally, a significant inverse relationship between the ability of L. paracasei BRAP01 and L. rhamnosus AD500 to stimulate IFN-γ/IL-10 or IL-10/IFN-γ production was also observed. CONCLUSIONS: Our results indicated that age and gender have only minor effects on the immune modulation abilities of probiotics.

Roles of the Nrf2/HO-1 pathway in the anti-oxidative stress response to ischemia-reperfusion brain injury in rats.

OBJECTIVE: The aim of this study was to investigate the roles of the Nrf2/HO-1 pathway in the responses to the oxidative stress created by ischemia-reperfusion brain injury in rats. MATERIALS AND METHODS: 54 healthy, adult, male SD rats were included in the study. Eighteen (18) rats were placed in the sham group. The ischemia-reperfusion model was created in the other 36 rats, among which 18 received injections of Nrf2 agonist before the surgery. The suture method was used to create artery occlusions in the right brain of the rats; and reperfusion was done after 90-minute ischemia (MCAO); while no suture was inserted in the sham group. At 3, 6, 12, 24, 48 and 72 hours after the modeling, their neurological functions were evaluated. Also, at different time points, rats were decapitated, and their fresh brain tissues were used to detect the infarct volume percentages by TTC staining and the brain water contents by the dry-wet weight method. The SOD contents in the brain tissue were measured by Xanthine oxidase assay. RT-PCR was used to detect the mRNA expression of HO-1 in the brain tissues, and western blot method was used to detect the expression level of HO-1 and Nrf-2. RESULTS: The rats in the sham group had no obvious neurological defects; while those in the MCAO group showed significant neurological defects at all time points. The MCAO group had higher neurological evaluation scores than the sham group. TTC staining showed that infarct in the MCAO group kept increasing over time and peaked at 24h. Measurements of SOD found that the sham group had the highest SOD among the three groups, and showed no significant fluctuation over time. The MCAO group had much lower SOD activities than the sham group at all the time points. The higher the level of HO-1mRNA and protein expression in the brain tissue of rats in each group, the higher the degree of brain injury, but the lower the level of Nrf2 protein expression and the lower degree of brain injury. Nrf2 agonist markedly improved all these indicators in the rats which underwent the MCAO surgery. CONCLUSIONS: The expression of HO-1 after ischemia-reperfusion brain injury may contribute to the increased infarct volume. Activation of Nrf2 could improve the prognosis of ischemia-reperfusion brain injury.

Anthocyanin contents in the seed coat of black soya bean and their anti-human tyrosinase activity and antioxidative activity.

OBJECTIVE: The seed coat of black soya bean (SCBS) contains high amount of anthocyanins and shows antioxidant and anti-mushroom tyrosinase activities. The objectives of this study were to analyse the anthocyanins in SCBS with different solvents and to find the relationship between anthocyanin profile with anti-human and anti-mushroom tyrosinase activities. METHODS: SCBS was extracted with hot water, 50 and 80% ethanol, 50 and 80% acetone and 50 and 80% acidified acetone. Total phenol and total flavonoid contents in the extracts were determined. Anthocyanins in the extracts were analysed using HPLC and LC/MS/MS. A genetically engineered human tyrosinase was used to evaluate the anti-tyrosinase potential of the extracts from SCBS. RESULTS: 80% acetone extract from SCBS obtained the highest total phenol, total flavonoid and cyanidin-3-O-glucoside (C3G) contents among all the extracts, whereas the hot water extract showed the lowest antioxidant contents. Three anthocyanin compounds were found in all the extracts from SCBS, and the analysis of HPLC and LC/MS/MS indicated that they were C3G, delphinidin-3-O-glucoside (D3G) and peonidin-3-O-glucoside (P3G). The ratios of C3G (2.84 mg g(-1) ), D3G (0.34 mg g(-1) ) and P3G (0.35 mg g(-1) ) in 80% acidified acetone extract were 76.6, 9.1 and 9.3%, respectively. All the extracts from SCBS possessed anti-human tyrosinase activity. Moreover, a good correlation was found between the anti-human tyrosinase activities and C3G contents in the extracts. CONCLUSION: Antioxidants in SCBS also possess anti-human and anti-mushroom tyrosinase activities.

Prognostic significance of p62/SQSTM1 subcellular localization and LC3B in oral squamous cell carcinoma.

BACKGROUND: Autophagy is a programmed cell survival mechanism that has a key role in both physiologic and pathologic conditions. The relationship between autophagy and cancer is complex because autophagy can act as either a tumour suppressor or as a tumour promoter. The role of autophagy in oral squamous cell carcinoma (OSCC) is controversial. Several studies have claimed that either a high or low expression of autophagy-related proteins was associated with poor prognosis of OSCCs. The aims of the study were to compare autophagy in OSCCs, verrucous hyperplasias, and normal oral mucosas, and to inspect the prognostic role of autophagy in OSCCs. METHODS: We used the autophagosome marker, LC3B, and autophagy flux marker, p62/SQSTM1 (p62), by using immunohistochemistry, and examined p62 mRNA by RNA in situ hybridization, to evaluate autophagy in 195 OSCCs, 47 verrucous hyperplasias, and 37 normal oral mucosas. The prognostic roles of LC3B and p62 protein expressions in OSCCs were investigated. RESULTS: We discovered that the normal oral mucosa exhibited limited LC3B punctae and weak cytoplasmic p62 staining, whereas the OSCCs exhibited a marked increase in LC3B punctae and cytoplasmic p62 expression. The expression pattern of LC3B and cytoplasmic p62 of the verrucous hyperplasias were between normal oral mucosas and OSCCs. The normal oral mucosas, verrucous hyperplasias, and OSCCs presented no differences in nuclear p62 expression and the p62 mRNA level. p62 mRNA expression was elevated in a minority of cases. High p62 mRNA expression was associated with high p62 protein expression in the cytoplasm. Increased LC3B punctae, high cytoplasmic p62, and low nuclear p62 expressions in OSCCs were associated with aggressive clinicopathologic features and unfavourable prognosis. In addition, low nuclear p62 expression was an independent prognostic factor for overall and disease-specific survival rates. Furthermore, we disclosed that high cytoplasmic p62 expression accompanied with either a low or high LC3B expression, which indicated autophagy impairment under basal or activated autophagic activity, was associated with aggressive behaviour in advanced OSCCs. CONCLUSIONS: We suggested that autophagy was altered during cancer initiation and progression. Autophagy impairment contributed to cancer progression in advanced OSCCs.

Celecoxib-induced increase in cytosolic Ca(2+) levels and apoptosis in HA59T human hepatoma cells.

Celecoxib has been shown to have antitumor effect in previous studies but the mechanisms are unclear. The effect of celecoxib on cytosolic Ca(2+) concentrations ([Ca(2+)]i) and viability in HA59T human hepatoma cells was explored. The Ca(2+)-sensitive fluorescent dye fura-2 was applied to measure [Ca(2+)]i. Celecoxib at concentrations of 10-50 µM induced a [Ca(2+)]i rise in a concentration-dependent manner. The response was reduced by 80% by removing Ca(2+). Celecoxib induced Mn(2+) influx, leading to quenching of fura-2 fluorescence. Celecoxib-evoked Ca(2+) entry was suppressed by nifedipine, econazole, SK&F96365, and protein kinase C modulators. In the absence of extracellular Ca(2+), incubation with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin nearly abolished celecoxib-induced [Ca(2+)]i rise. Incubation with celecoxib abolished thapsigargin-induced [Ca(2+)]i rise. Inhibition of phospholipase C with U73122 abolished celecoxib-induced [Ca(2+)]i rise. At 1-50 µM, celecoxib inhibited cell viability by less than 20%, which was not reversed by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N, N, N', N'-tetraacetic acid/acetoxy methyl (BAPTA/AM). Celecoxib (10-50 µM) also induced apoptosis. In sum, in HA59T hepatoma cells, celecoxib induced a [Ca(2+)]i rise by evoking phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via protein kinase C-sensitive store-operated Ca(2+) channels. Celecoxib also caused cell death via apoptosis.

AKT2 expression in histopathologic grading and recurrence of meningiomas.

BACKGROUND: AKT2 (protein kinase B), an important protein in PI3K signaling pathway, is overexpressed in a variety of malignant tumors. However, in patients with meningiomas, the potential correlation between AKT2 and clinical outcome remains unknown. METHODS: The expression of AKT2 and Ki-67 in meningioma tissues were evaluated immunohistochemically in 94 patients with meningiomas. The correlation of AKT2 immunoreactivity with clinicopathological features and the prognostic value of AKT2 in patients were also analyzed. RESULTS: In this study, we examined the expression of AKT2 in meningiomas and unveiled its possible relationship with the clinical outcome. Immunohistochemical analysis revealed high AKT2 expression in 46 patients (46/94, 48.9%) and low AKT2 expression in the remaining 48 patients (48/94, 51.1%). There was a positive correlation between AKT2 and Ki-67 immunoreactivity (r = 0.35, P = 0.01). Clinicopathological evaluation suggested that AKT2 expression was associated with pathological grade and recurrence (P < 0.05). Univariate and Cox analysis indicated a significant correlation between high levels of AKT2 immunoreactivity and high rates of tumor recurrence (P < 0.05). CONCLUSIONS: We conclude that AKT2 may play an important role in the development of meningioma. High AKT2 labeling index indicates higher grade of meningioma, and therefore AKT2 may be a useful molecular marker for predicting the prognosis of meningioma.

Grp78 as a therapeutic target for refractory head-neck cancer with CD24(-)CD44(+) stemness phenotype.

Cancer stem cells are refractory to conventional therapy, which result to cancer metastasis and chemo-radioresistance. Grp78 is known to have important roles in cytoprotection and tumorigenesis in several cancers. We therefore examined whether Grp78 can serve as a therapeutic target for refractory stemness phenotype of head and neck cancer (HNC). Six HNC cell lines were used. Fluorescence-activated cell sorting (FACS) analysis was used to sort CD24(-)CD44(+) and Grp78(+) cells. The small interfering RNA (siRNA) knockdown and cDNA transfection were applied to examine the effects of Grp78 on cellular function. Western blot and confocol microscopy were used to determine the effects of downstream protein expressions. Xenografted mouse tumors and immunohistochemistry were used to validate the results. We found that Grp78 regulated the conversion of CD24(-)CD44(+) cells, a characteristic of HNC stem cells. The CD24(-)CD44(+)Grp78(+) cells showed superior chemo-radioresistance and invasion ability compared with CD24(-)CD44(+), Grp78(+) or the parental cells. Silencing Grp78 increased chemo-radiosensitivity, inhibited cell invasion, reverse epithelial-mesenchymal transition, suppressed cancer stemness, withdrew CD24(-)CD44(+) cell conversion and induced differentiated phenotype. Study in xenografted mice further showed that CD24(-)CD44(+)Grp78(+) cells exhibited highest tumorigenesis, compared with CD24(-)CD44(+) CD24(+)CD44(+) or the parental cells. Grp78 knockdown dramatically restrained tumor growth along with the inhibition of stem cell regulatory proteins Oct-4 and Slug. Grp78 may serve as a molecular target that can be further developed for eradication of refractory HNC with stemness phenotype.

Effect of diindolylmethane on Ca(2+) homeostasis and viability in MDCK renal tubular cells.

The effect of the natural product diindolylmethane (DIM) on cytosolic Ca(2+) concentrations ([Ca(2+)]i) and viability in MDCK renal tubular cells was explored. The Ca(2+)-sensitive fluorescent dye fura-2 was applied to measure [Ca(2+)]i. DIM at concentrations 1-50 µM induced a [Ca(2+)]i rise in a concentration-dependent manner. The response was reduced partly by removing Ca(2+). DIM induced Mn(2+) influx leading to quenching of fura-2 fluorescence. DIM-evoked Ca(2+) entry was suppressed by nifedipine, econazole, SK&F96365 and protein kinase C modulators. In the absence of extracellular Ca(2+), incubation with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin (TG) or 2,5-di-tert-butylhydroquinone (BHQ) greatly inhibited DIM-induced [Ca(2+)]i rise. Incubation with DIM abolished TG or BHQ-induced [Ca(2+)]i rise. Inhibition of phospholipase C with U73122 reduced DIM-induced [Ca(2+)]i rise by 50%. At 1, 10, 40 and 50 µM, DIM slightly enhanced cell proliferation. The effect of 50 µM DIM was reversed by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid. In sum, in MDCK cells, DIM induced a [Ca(2+)]i rise by evoking phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via protein kinase C-sensitive store-operated Ca(2+) channels. DIM did not induce cell death.

Evaluation of discrepancies between oxacillin and cefoxitin disk susceptibility for detecting methicillin-resistant Staphylococcus aureus.

The Clinical Laboratory Standards Institute recommends that if both cefoxitin and oxacillin are tested against Staphylococcus aureus and either result is measured as resistant, the organism should be reported as oxacillin resistant. This indicates that discrepancies may be present between oxacillin and cefoxitin sensitivities in S. aureus. In this study, we aimed to investigate the discrepancy between oxacillin and cefoxitin susceptibility in S. aureus clinical isolates. Of 10,980 S. aureus isolates recovered from 2005 to 2010, 27 (0.3%) isolates with discordant results between oxacillin and cefoxitin were collected. Fourteen (oxacillin diameters 10-12 mm) of the 27 strains were susceptible (MICs = 0.5-2 µg/ml) and 13 (6-13 mm) were resistant (4->256 µg/ml) to oxacillin. The cefoxitin MICs of 14 oxacillin-susceptible and 13 oxacillin-resistant strains ranged between 4 and 8 and 8 to 32 µg/ml, respectively. Discrepancies were present between oxacillin and cefoxitin in S. aureus, and these strains should be further tested for oxacillin MICs and for the mecA gene or ß-lactamase activity.

Evaluation of discrepancies between oxacillin and cefoxitin susceptibility in coagulase-negative staphylococci.

The Clinical and Laboratory Standards Institute (CLSI) recommends testing coagulase-negative staphylococci (CoNS) strains to determine resistance against oxacillin by testing for mecA, PBP2a, or with cefoxitin disk. However, discrepant results of resistance to oxacillin and susceptibility to cefoxitin were found. In this study, we aimed to investigate the oxacillin resistance and cefoxitin susceptibility of CoNS in Taiwan. Of 9,017 strains collected from 2005 to 2010, 131 (1.5%) of the isolates were oxacillin-resistant and cefoxitin-susceptible. Species identification was carried out using the Vitek 2 system or 16S ribosomal RNA sequencing. Oxacillin minimum inhibitory concentrations (MICs) were examined by the agar dilution method. The presence of mecA and the activity of ß-lactamase were performed by polymerase chain reaction (PCR) and Cefinase disks, respectively. Overall, 33% (43/129) of the strains carried mecA and 43% (37/86) of mecA-negative isolates tested positive for ß-lactamase. The remaining 49 isolates were negative for both mecA and ß-lactamase, and were mainly Staphylococcus cohnii ssp. urealyticus and S. saprophyticus (oxacillin MICs 0.5-2 µg/ml) obtained from bloodstream and urinary tract infections. Our study suggests that incorrect reporting can be found in CoNS using cefoxitin disk alone to determine the susceptibility to oxacillin, and the strains should be further tested for oxacillin MICs and detection of the mecA gene or ß-lactamase activity.

Highly potent and specific siRNAs against E6 or E7 genes of HPV16- or HPV18-infected cervical cancers.

Infection with high-risk types (type 16 or type 18) of human papillomaviruses (HPVs) increases a patient's risk of cervical cancer. Given the importance of the cervix and the severe side effects resulting from traditional cancer therapies, this study aimed to achieve targeted inhibition of viral oncogenes in tumor cells using small interfering RNAs (siRNA). To accomplish this, we developed nine siRNAs against either the E6 or E7 genes of HPV-16 or HPV-18 in several combinations, yielding siRNAs targeting 16E6, 16E7, 18E6 and 18E7. We measured the effectiveness of the siRNAs by examining E6 or E7 mRNA expression after transfection of the siRNAs into HPV-positive CaSki (HPV-16) or HeLa (HPV-18) cell lines. We found that the HPV-siRNAs significantly reduced cell growth and colony formation in both cell lines. Flow cytometry analysis revealed a significant increase in apoptosis. The siRNAs had no effect on cell growth, colony formation or apoptosis in HPV-negative C33A cells, demonstrating a lack of off-target effects. In addition, an in vivo xenograft study showed that intra-tumoral injection of the siRNAs reduced tumor growth in BALB/c nude mice. In conclusion, we have developed highly specific and potent HPV-siRNAs that successfully suppress tumor growth and induce apoptosis in HPV-positive cervical cancer cells. siRNA treatment has potential for further development as an adjuvant therapy for cervical cancer.

Impact of food and antacids on the pharmacokinetics of anti-tuberculosis drugs: systematic review and meta-analysis.

OBJECTIVE: To compare the effects of food and antacids on the bioavailability of first-line anti-tuberculosis drugs. METHOD: Systematic search of electronic databases PubMed (January 1950-May 2009), and the Cochrane Library database (January 1974-May 2009), including the Cochrane Centre register of controlled trials, and ongoing trials from research registers using key terms 'food', 'antacids', 'meal', 'controlled trial', 'diet', and the first-line anti-tuberculosis drugs isoniazid (INH), rifampicin (RMP), ethambutol (EMB) and pyrazinamide (PZA). Meta-analysis was performed using RevMan software 5 to assess the impact of food or antacids on the maximum plasma concentrations (C(max)) and area under the plasma concentration time curve (AUC) of anti-tuberculosis drugs. RESULTS: Twelve trials involving 157 patients were included in the meta-analysis. The overall effects showed that food significantly reduced the C(max) mean difference (C(max) MD; C(max) MD -1.42, 95%CI -1.56--1.28, P < 0.00001) and AUC (C(max) MD -3.33, 95%CI -4.05--2.62, P < 0.00001) of INH but antacids did not. Food also significantly reduced the C(max) MD (C(max) MD -2.47, 95%CI -3.30--1.64, P < 0.00001) but not the AUC of RMP. Antacids had no effect on the C(max) MD or AUC of RMP. The C(max) and AUC of PZA were unaffected by both food and antacids. Both food and antacids reduced the C(max) but not the AUC of EMB. CONCLUSION: From a pharmacokinetic point of view, it seems that the better option for patients with gastrointestinal upsets during chemotherapy would be to add antacids rather than dosing with meals.