It has been previously shown that the double PRLRKO/ D2RKO mice present more marked pituitary hyperplasia and increased pituitary weight compared to the single D2RKO counterpart. Our results may explain why abrogation of PRLR signaling in PRLKO or PRLRKO mice contributes to pituitary hyperplasia. PRLRKO female mice develop pituitary hyperplasia from 6 months of age, whereas male mice do not show pituitary enlargement until 18 months of age. In females, the apoptotic and proliferative rates are acutely and closely regulated at each stage of the estrous cycle. Our results indicate that in PRLRKO female mice there is a progressive and accumulative effect due to the lack of PRL apoptotic and antiproliferative actions in each proestrus. These results also could explain why PRLRKO female mice develop pituitary hyperplasia earlier than males, although PRL also plays a key role in regulating pituitary remodeling in males. Based on the evidence provided here that PRL, like dopamine, regulates anterior pituitary homeostasis by controlling cell proliferation and apoptosis, our findings further support the earlier hypothesis of Schuff et al that a deficiency of these two individual pathways results in additive effects on the development of pituitary tumors in double PRLRKO/D2RKO mice. In accordance with this hypothesis, we also observed that chronic blockade of PRLR by the antagonist D1-9-G129R-hPRL increased the pituitary weight in TG female mice. Based on these results, we propose a general model depicted in Figure 8. At proestrus, the high circulating levels of estrogens upregulate PRL secretion which in turn, together with estradiol itself, induces apoptosis of anterior pituitary cells. Furthermore, PRL stimulates dopamine release from hypothalamic dopaminergic neurons to the portal vessels. Dopamine reaching the anterior pituitary could also contribute to the apoptosis observed in the gland at this stage of the estrous cycle. Estradiol also increases anterior pituitary PRL cleavage into 16 kDa N-terminal fragments, which in turn induce apoptosis in an estrogen-dependent manner, by a PRLRindependent mechanism. The molecular mechanisms by which PRL regulates pituitary homeostasis during the estrous cycle are unknown. Since different PRLR isoforms exhibit different capabilities to activate intracellular signaling pathways, the ultimate effects of PRL depend not only on the number, but also on the type of receptor isoforms expressed in target tissues. It is thus important to determine the pattern of PRLR expression before moving further in intracellular signaling investigations. In the present study we show that the anterior pituitary from female mice expresses short and long PRLR isoforms and that expression of both are increased at proestrus. This suggests that PRLR expression in the anterior pituitary is regulated by circulating hormones whose levels fluctuate during the estrous cycle.

The pioneering work on the influence of age and sex on collagen autofluorescence was reported earlier by Shuster and co-workers involving human subjects which concluded that skin collagen decreases with age. On the contrary, in a study by Kollias and co-workers, a positive correlation between age and collagen deposition by measuring endogenous skin fluorescence using albino hairless mouse models was observed. In the present study although, there were variations in the collagen intensities among un-illuminated control and laser treatment groups with age, it was found to be nonsignificant, may be attributed to the selected age range, which would have been improved if the age range selected was between 5-55 weeks. Similarly, autofluorescence results indicated higher collagen deposition in male mice compared to females in all the tested time points, however the difference in collgen deposition was not found to be statistically significant. Present report is the first of its kind, wherein autofluorescence technique has been utilized to monitor collagen non-invasively as well as to reveal the differential collagen deposition with gender during acute wound healing. Previous studies from our laboratory contributed significantly in demonstrating the augmented effect of red light laser irradiation on collagen synthesis in pre-clinical models during tissue regeneration. Thus, the major focus of the current work was to elucidate the prognostic potential of in vivo autofluorescence in the factual measurement of collagen rather than proving the beneficial effect of LLLT in collagen deposition. In order to achieve the stated objective, normalized spectral intensity and area under the curves for collagen peaks were computed and compared. In our previous report, we have utilized normalized NADH/collagen ratio to assess the collagen deposition in the spectra. However, in the present study we believed that the inclusion of collagen intensity and an additional parameter such as AUC could help in acquiring more accurate information about the collagen. Although, collagen intensities and AUC were extracted from the same spectra, we believe that the intensity could provide information at a single point i.e., at a wavelength of 405 nm whereas, AUC could provide supplementary information to the spectral intensity values. Thus, AUC measurments in the present study were performed only to support/confirm the findings of collagen intensity at 405 nm. Single exposure of the optimum laser treated animals displayed 1.171, 1.165, 1.125, 1.020 and 1.012 fold increase in collagen intensity and 1.165, 1.151, 1.127, 1.019 and 1.016 fold increase in AUC values compared to un-illuminated controls on days 5, 10, 30, 45 and 60 post-wounding respectively. Two different spectral parameters i.e. spectral intensity and AUC displayed similar and comparable value for collagen deposition substantiating.

In wheat, plastidial ADP-glucose transporter was also analyzed by reconstituting amyloplast envelope membrane proteins in proteoliposomes. Its apparent affinities for ADP-Glc, and both ADP and AMP were found to be 430 mM and 200 mM, respectively. Enhancing the efflux of ADP-Glc by dilution of the medium with high concentration of ADP and AMP revealed the potentials of these nucleotides as counter exchange substrates. This result was supported by the efflux of the putative ADP by dilution with high concentration of non-labeled ADP-Glc. In agreement with this, efflux study with intact amyloplasts of wheat showed that ADP-glucose transporter protein transports ADP-Glc into the amyloplasts in counter exchange with ADP and AMP. Colorectal cancer is the third most diagnosed cancer type in males and the second in females worldwide, and its incidence is increasing even in traditionally low-risk countries such as Spain. Moreover, mortality rates caused by colorectal cancer remain high, being the fourth and third cause of cancer-related mortality in males and females, respectively. On the other hand, pancreatic cancer affects 250 000 individuals worldwide annually. Although its incidence rates are not very high, it is one of the most lethal tumors, representing the five and fourth cause of cancer-related mortality in males and females, respectively, in developed countries. Therefore, new complementary therapeutic approaches, ideally cost-effective and non-toxic, are needed to improve efficacy and quality of life of patients with these cancer types. Rosemary and many of its components were reported to possess chemopreventive properties in skin and breast cancers in vivo, mostly by inhibiting 7,12dimethylbenzanthracene -DNA adduct formation. Moreover, they exert antioxidant activity both in vitro and in vivo, thus inhibiting genotoxicity, which is a significant contributory cause of cancer, and protecting from carcinogens or toxic agents. They also were reported to display antiproliferative activity in vitro against breast, leukemia, hepatoma, colon, lung, prostate, ovarian, and urinary bladder cancer cells. However, the effect of rosemary on pancreatic carcinoma cells has not been reported to date. Regarding the tumor progression in vivo, the effect of rosemary extract in combination with an analogue of 1,25-dihydroxyvitamin D3 was assessed in a syngeneic mouse leukemia tumor model, and showed a strong cooperative antitumor effect. Several rosemary components, such as carnosic acid, carnosol, ursolic acid, as well as some of its essential oil constituents, have been proposed to be responsible for the anticancer effects of rosemary extracts. Although the concentration ratios of carnosol and carnosic acid were reported to influence the antioxidant and antimicrobial activities, the possible synergism of the rosemary components regarding the antitumor activity of rosemary extracts has not been reported yet.

The encouraging results presented in this report will support our consideration for further development of this novel therapeutic duplet in the treatment of patients with NMIBC. After 1 hour, the purse string suture was removed and rats were stimulated to expel bladder contents. The intravesical therapy was administered weekly for a total of six weeks to mimic intravesical BCG therapy in humans. Two weeks after completion of the intravesical therapy, animals were sacrificed and tissues were harvested and processed for subsequent analysis. Figure 1 illustrates the study treatment schema. As previously described, the IL-15N72D mutein was generated through site-directed mutagenesis of IL-15 wild type and found to exhibit super agonist activity through improved interactions with the human IL-15Rb chain. The IL-15N72D mutein displayed,4 fold lower effective concentration than wildtype IL-15 and the super agonist activity was enhanced further by pre-association with either mouse or human IL-15Ra-Fc molecules. Overall, the IL-15N72D/IL-15Ra-Fc complex exhibited a 25-fold lower EC50 than IL-15 alone for supporting IL-15-dependent cell growth, indicating the complex provides highly potent biological activity. The enhanced activity of the IL-15N72D/IL-15Ra-Fc complex is likely the result of a combination of the increased binding activity of the N72D mutein to the IL-2Rbc complex, optimized cytokine trans-presentation by the IL-15Ra chain, the dimeric nature of the cytokine domain and its increased in vivo half-life compared to native IL-15. Recently, Xu et al. reported on the therapeutic efficacy of intravenous ALT-803 in a myeloma xenograft model and alluded to the possible mechanism behind ALT-803 activity. Briefly, a single dose of ALT-803, but not IL-15 alone, eliminated wellestablished 5T33P and MOPC-315P myeloma cells from the bone marrow of tumor-bearing mice and prolonged survival in myeloma-bearing mice. ALT-803 treatment stimulated CD8 cells to secrete IFN-c and promoted rapid expansion of CD8CD44 memory T-cells. These memory CD8 T- cells also exhibited nonspecific cytotoxicity against myeloma and other tumor cells in vitro, whereas IFN-c had no direct effect on myeloma cell growth in vitro. Though ALT-803 lost its antimyeloma activity in tumor-bearing IFN-c knockout mice, it was able to retain its ability to promote CD8CD44 memory T-cell proliferation, indicating that ALT-803-mediated stimulation of CD8CD44 memory T-cells is IFN-c-independent. ALT-803 stands out as a potent immunostimulant that is capable of simultaneously activating the innate and adaptive arms of the immune system to elicit both rapid and long-lasting protective responses against infectious or neoplastic challenges to the host. Thus, we hypothesize that intravesical administration of ALT-803 plus BCG will provide durable and potent cell-mediated immune responses. In the current study, we confirmed that ALT-803 reduction in angiogenesis.

PKM2 in cancer cells, such as the human lung cancer cell line H1299, and replacing it with PKM1 was demonstrated to result in a metabolic phenotype change involving decreased glucose uptake and increased oxidative phosphorylation. Further, reprogramming of somatic cells to induced pluripotent stem cells has also been shown to incur metabolic reprogramming; the change from a low glycolytic flux state of somatic cells to a high flux state of rapidly dividing iPSC cells is accompanied by a switch in the isozyme expression of HK and PFK enzymes. An additional layer of flux regulation of glycolysis is exerted by signaling pathways. Through signaling pathways, contrasting glycolysis flux behavior is accomplished without changing the isoforms ; instead the action of signaling pathways alters the kinetic behavior of the target enzyme. Tyrosine kinase signaling has been shown to change the kinetic behavior of PKM2 isoform through modulation of its allosteric regulations. Similarly, signaling events triggered by glucagon in hepatocytes alter the kinetics of the liver isozyme of PK. Protein kinases A/B/C have been shown to affect the kinetics of PFKFB isoforms. The composition of isozymes in glycolysis, through multiple layers of regulation, is pivotal to the flux control and plays a key role in growth control and physiological balance. Over the last four decades, the kinetic behavior of isoforms of individual glycolytic enzymes has been examined in detail. However, a holistic understanding of the effect of different combinations of such isoforms on the flux behavior of the complete glycolysis pathway is yet to be attained. We have taken a systems biology approach to study the flux states of glycolysis pathway. Using a mathematical model that employs mechanistic rate equations for enzyme kinetics, we demonstrate that glycolysis exhibits a classical multiple steady state behavior in terms of its flux with respect to the glucose concentration. The multiplicity of steady states segregate cell metabolism into distinct states: high glycolytic flux states and low glycolytic flux states. Such bistable behavior is an output of complex allosteric regulations which in turn depend on the type of glycolytic isozymes expressed. We show that the presence of the muscle or the liver isozyme of PFK or/and the L, R or M2 isoform of PK is necessary for multistability in glycolytic flux. We substantiated the modeling insights with gene expression data from various tissues as well as experimental data from HeLa cells. Further, we discuss the factors that affect the bistable nature of the glycolysis such as the level and the K/P ratio of enzyme PFKFB. Similar kinds of bistable behavior have been shown to act like a robust switch in many regulatory circuits including oocyte cell maturation, transition between multiple phosphoform stable states in multisite phosphorylation systems, among many others.