In Vitro Regulation of Cell Growth and Angiogenesis by Inositol Hexaphosphate in Bladder Cancer

Background

Inositol Hexaphosphate (IP6) is a naturally occurring polyphosphorylated carbohydrate that is found in food sources high in fiber content. We hypothesized that IP6 would inhibit the cell growth rate of bladder cancer in vitro.

Methods

T24 and TCCSUP bladder cancer cell lines were treated with titrating doses of IP6 (0.3, 0.6 and 0.9 mM/well). Cell viability and vascular endothelial growth factor levels were measured.

Results

Significant reductions (p < 0.001) in cellular growth were noted in both cell lines at all doses and time points tested, with the exception of 0.3 mM IP6 at 24 hours in the T24 cell line. The percent inhibition of vascular endothelial growth factor was significantly higher than that observed in the TCCSUP cell line at 48 and 72 hours with 0.3 mM IP6 (p < 0.001). The T24 cells exhibited the same level of inhibition at 24 and 48 hours with 0.6 mM dose of IP6 and at 72 hours with the 0.3 mM dose (p < 0.001).

Conclusions

In vitro treatment of bladder cancer with the common dietary polyphosphorylated carbohydrate IP6 significantly decreased cellular growth by anti-angiogenic mechanisms. We feel that this data warrants further investigation and consideration for initiation of clinical trials to evaluate the safety and clinical utility of this agent.Key Words: Cellular anti-proliferation, Bladder cancer, Angiogenesis, Inositol hexaphosphate, Vascular endothelial growth factor     

Coping with Stressful or Traumatic Events: What Aspects of Trauma Reactions are Associated with Health Outcomes?

The presence of posttraumatic stress disorder (PTSD) symptoms has been shown to be related to a number of health outcomes. In the current study, we explored which specific aspects of PTSD are most related to health measures. The associations between the specific DSM‐IV‐TR PTSD criteria (criteria A–F) and five indicators of health and well‐being—physical health symptoms, quality of life, mental health, depression and negative affect—were examined. The sample consisted of 711 undergraduates. A non‐clinical sample was recruited so there would be variability in the various criteria for PTSD. Multiple regression analyses revealed that the hyperarousal (criterion D) was the most consistent and strongest predictor of outcomes. However, the F criterion (causes significant impairment) predicted additional variance in quality of life, depression and negative affect. These results suggest that it is not just the mere frequency of trauma symptoms that affect well‐being but also the disruptive capability of these symptoms. In addition, follow‐up analyses indicated that hyperarousal mediated the association between the A2 criterion (traumatic response) and all five outcome measures. These results underscore the importance of the hyperarousal criterion, while also suggesting the need for increased attention to the F criterion when considering the impact of stressful events on health and well‐being. Copyright © 2012 John Wiley & Sons, Ltd.     

Complementary suppression of T cell activation by peritoneal macrophages and CTLA-4-Ig

Peritoneal macrophages suppressed T lymphocyte activation by amino acid catabolism. CTLA-4-Ig complemented this form of suppression by blocking the costimulation of T cells. Inhibition of the amino acid catabolizing enzymes or blockade of the IFNgamma signaling essential for macrophage-mediated suppression did not impact CTLA-4-Ig efficacy. Except for phytohemagglutinin, T cell responses to superantigen, mitogen, and anti-CD3 were further reduced when CTLA-4-Ig was combined with peritoneal macrophages. The combination of these complimentary forms of immune regulation potently suppresses adaptive immunity.     

Exposure?response modelling for empagliflozin,a sodium glucose cotransporter 2 (SGLT2) inhibitor,in patients with type 2 diabetes

Aims

To provide model-based clinical development decision support including dose selection guidance for empagliflozin, an orally administered sodium glucose cotransporter 2 inhibitor, through developed exposure−response (E−R) models for efficacy and tolerability in patients with type 2 diabetes mellitus (T2DM).

Methods

Five randomized, placebo-controlled, multiple oral dose studies of empagliflozin in patients with T2DM (n = 974; 1–100 mg once daily, duration ≤12 weeks) were used to develop E−R models for efficacy (glycosylated haemoglobin [HbA_1c], fasting plasma glucose [FPG] and urinary glucose excretion). Two studies (n = 748, 12 weeks) were used to evaluate tolerability E−R.

Results

The efficacy model predicted maximal decreases in FPG and HbA_1c of 16% and 0.6%, respectively, assuming a baseline FPG concentration of 8 mm (144 mg dl⁻¹) and 10–25 mg every day empagliflozin targeted 80–90% of these maximums. Increases in exposure had no effect on incidence rates of hypoglycaemia (n = 4), urinary tract infection (n = 17) or genital/vulvovaginal-related (n = 16) events, although low prevalence rates may have precluded more accurate evaluation.

Conclusions

E−R analyses indicated that 10 and 25 mg once daily empagliflozin doses achieved near maximal glucose lowering efficacy.     

Modulation of transforming growth factor-beta production in normal human osteoblast-like cells by 17 beta-estradiol and parathyroid hormone

Although our laboratory has reported that normal human osteoblast-like (hOB) cells contain estrogen receptors, we have failed to find major effects of 17 beta-estradiol (E2) on modulation of proliferation of bone matrix protein production by hOB cells. Because the major effect of E2 in vivo is to decrease bone resorption and because transforming growth factor-beta (TGF-beta) has been reported to decrease osteoclast-mediated bone resorption, we have tested the hypothesis that the effect of E2 on osteoclast activity is, at least in part, indirectly mediated by enhancing production of TGF-beta by osteoblasts. We therefore have extended our studies to examine possible TGF-beta gene expression including the modulation of the release of TGF-beta by E2 in near homogenous populations of hOB cells. TGF-beta protein production was measured using growth inhibition of CCL-64 cells and verified by blocking effects with anti-TGF-beta antibodies. TGF-beta 1 messenger RNA (mRNA) steady state levels were assessed by northern blot analysis and quantitated by densitometric measurement using 18S ribosomal RNA as a reference. There was an E2 dose-dependent increase in TGF-beta protein production within 24 h of challenge with E2. Northern blots from these cells demonstrated a dose-dependent increase in steady state mRNA levels of TGF-beta 1 within 6 h of treatment. PTH was also a potent stimulator of TGF-beta protein and message levels in a dose-dependent manner. Interestingly, coincubation of equimolar concentrations of E2 and PTH (10(-8) M) abrogated the stimulation of TGF-beta 1 mRNA and protein. Decreasing the relative concentration of PTH in this coincubation with E2 increased TGF-beta 1 mRNA and protein levels. These data support the fact that E2 modulates TGF-beta production in osteoblasts. In this manner TGF-beta may mediate E2 inhibition of osteoclast activity.     

Bone formation rate in older normal women: concurrent assessment with bone histomorphometry, calcium kinetics, and biochemical markers

In 12 younger (age, 30-41 yr) and 11 older (age, 55-73 yr) normal women we assessed bone formation rate using multiple methods. Bone formation (mean +/- SE) was higher in the older women than in the younger women, based on measurements of serum bone Gla-protein (1.67 +/- 0.07 vs. 1.14 +/- 0.10 nmol/L; P less than 0.01), serum bone-specific alkaline phosphatase activity (388 +/- 42 vs. 223 +/- 22 nanokatal/L, P less than 0.01), and bone formation rate by histomorphometry of iliac biopsy (31.1 +/- 4.9% vs. 15.1 +/- 2.7%/yr; P less than 0.01), but was similar in the two groups when accretion rates were assessed by calcium kinetics (5.9 +/- 1.0 vs. 7.5 +/- 1.2; P = NS). This latter discrepancy may have been caused by several age-related factors, especially reduced mineralization of completed osteons, and by not correcting for the decrease in total skeletal calcium in the older group. Our data call into question the traditional belief that bone turnover decreases in older women.