Involves nucleotide binding and hydrolysis, but how this occurs is not well understood. Septins polymerize by interacting with one another via the N- and C-termini and the nucleotide-binding pocket of their GTP binding domains. Structural studies indicate that GTP-binding triggers conformational changes that destabilize the NC interface, while GTP hydrolysis appears to have the opposite effect on the G interface, which is more stable in its GDP- than GTP-bound state. Septin monomers hydrolyze GTP faster than septin dimers and oligomers, whose nucleotide-binding pocket is not readily accessible by GTP. Not all septins, however, are capable of hydrolyzing GTP. Septins that lack a threonine residue, which is critical for GTP hydrolysis, are constitutively bound to GTP and septin hetero-hexamers contain both GDP and GTP at a ratio of 2:1. Interestingly, in vitro biochemical studies of septin assembly have shown that GTP-gamma-S, a slowly hydrolysable GTP analog, promotes the assembly of septin monomers into homo-polymeric filaments. While more work is needed to fully understand how GTP-binding and hydrolysis affects the dynamics of septin filament assembly, pharmacological agents that stabilize or depolymerize septin filaments can be useful tools in understanding the mechanisms of septin assembly and function. To date, forchlorfenuron -N9phenylurea; CPPU) is the only small molecule compound known to affect septin filament assembly. FCF is a synthetic plant cytokinin that consists of a chlorinated pyridine and a phenol ring joined together by a urea group. FCF was fortuitously discovered to reversibly affect the localization and morphology of septins in the budding yeast Saccharomyces cerevisiae. Subsequent studies in mammalian cells and the filamentous fungus Ashbya gossypii showed that FCF dampens the dynamics of septin filaments, amplifying the length and thickness of septin filaments. Notably, FCF had a similar effect in a cell-free in vitro assay, boosting the assembly of purified recombinant septin complexes into higher order filamentous structures. While not ruling out the possibility of off-target effects in cells, these studies suggest that FCF has a direct effect on septin assembly. Understanding how FCF binds and affects septins can provide new insights into the mechanism of septin polymerization and guide the design of small molecule compounds that target septins with high specificity and affinity. Computational simulations of drug-target interactions using in silico molecular docking and molecular dynamics approaches are commonly used for the rational design and screening of drugs. As evidenced by the paucity of high-resolution atomiclevel septin structures, studying septin-FCF interactions by X-ray crystallography can be very challenging. To gain an insight into how FCF binds septins, we performed in silico simulations of FCF docking to all available high-resolution crystal structures of septins.

The ultimate goal of our research was to document for others the potential sources of accessible internal company data for public health and healthcare research, particularly in the area of pharmaceutical research, and, in doing so, to take the first steps toward exploring the current use and future potential for repositories of internal company information. Internal company documents serve as a valuable source of information about industry activities for those who wish to know about the impact of those activities upon the health of the public. Internal documents from pharmaceutical companies include not only information on marketing and policy activities but also contain quantitative and other data related to clinical trials carried out on company products. Data from all trials are critical for a complete and accurate assessment of interventions within systematic reviews. In response to this imperative, systematic reviewers and other healthcare stakeholders have been working hard towards making trial data held by companies publically available so that patients, providers, and policy makers can have a full picture of all that is known. Because internal company documents are not located in a single place, and they are not published or indexed in a bibliographic database, they are difficult to identify and locate. We elected to identify research articles using internal company documents across all industries as a starting point, knowing that we would likely leave many documents that are available for research unidentified. What we learned, first, is that thousands of internal tobacco company documents, mainly released through litigation, are located in repositories around the world, including searchable online repositories. Ninety percent of the research articles meeting our criteria used internal tobacco company documents and all but one of these used documents made available through litigation. The reference standard we used for articles employing internal tobacco documents indicated that our searches failed to find hundreds of additional, potentially eligible, studies of tobacco documents and that our searches also identified tobacco articles not included in the Tobacco Documents Library. The second thing we learned was that identification of nontobacco studies using internal company documents was harder than we had anticipated. Only 36/361 articles that we identified used non-tobacco sources, and more than half of these were concerned with pharmaceutical company documents. We made every effort to ensure a thorough search of PubMed and Embase databases to retrieve all relevant documents, and to be practical we designed a search strategy that elected precision over sensitivity. It is possible that there are additional relevant articles from other non-tobacco industries that our search failed to retrieve. We need to identify better search terms for retrieving articles that use internal company document.

In this study, we attempted to use lactoferrin to improve HFCSinduced HMMS including hepatic steatosis, insulin resistance, inflammation and oxidative stress in a murine model. In the western diet, long-term consumption of beverages containing HFCS contributes to the development HMMS, including obesity, insulin resistance, hypertriglyceridemia, NAFLD, and NASH, all of which are associated with an inflammatory state. Previous studies have indicated that administration of 15-54% aqueous solutions of HFCS can elevate fasting blood glucose concentrations and increase insulin resistance in murine models. Inflammatory status indicates the progression of obesity, insulin resistance, NAFLD, and NASH. In this study, gross examination of the livers of HFCSadministered mice showed lipid accumulation and a 45% increase in liver weight. Microscopic examination of liver sections revealed steatosis after HFCS administration, which was significantly reduced in a dosedependent manner by lactoferrin treatment. HFSC stimulates overgrowth of intestinal microbiota, increasing intestinal permeability and leading to chronic inflammation. In this study, HFCS induced overgrowth of fecal coliforms and higher serum LPS. Our data indicate that lactoferrin altered the HFCS-induced imbalance in intestinal microbiota and reduced chronic inflammation. In previous studies, lactoferrin presents to exert anti-infectious and anti-inflammatory activities in vivo, and to inhibit LPS-induced IL6 secretion in a human monocytic cell line. This observation was extended with a demonstration that lactoferrin inhibits the expression of mRNA of proinflammatory cytokines, including TNF-a, IL-1, IL-6, and IL-8, and modulates the nuclear transcription factor kappa B signaling cascade. Lactoferrin has also been shown to downregulate IL-10 secretion in LPS-stimulated macrophages. In our study, serum and hepatic LPS levels significantly increased in HFCS-induced HMMS, and this increase was significantly reduced IL-10 by lactoferrin. Significantly, reduced expression of TLR-4 was also observed in lactoferrin-treated groups indicated the reducing inflammatory cascade signaling. Puddu et al., indicate bovine lactoferrin conteracts TLR mediated activation signals in monocyte-derived dendritic cells. Serum ALT is a direct indicator of hepatitis, and lactoferrin reduced sALT in a dose-dependent manner. Furthermore, cytokine measurements indicated that lactoferrin could significantly reduce hepatic IL-1b, TNF-a, and MCP-1. Previous studies have shown that lactoferrin can bind LPS from E. coli and Salmonella typhimurium and remove the glycolipid from the bacterial surface. Lactoferricin, a peptide produced by hydrolysis of lactoferrin by a gastrointestinal digestive enzyme, plays a central role in scavenging LPS. Analysis of the structural characteristics of lactoferricin identified a six-residue sequence responsible for binding LPS of E. coli or Pseudomonas aeruginosa.

Perlucin from H. laevigata was first purified and characterised as a water-soluble protein of 155 amino acids from the nacreous layer. Based on the amino acid sequence a synthetic gene has been designed, although the natural coding sequence has not yet been determined. Perlucin comprises a C-type lectin with a lectin domain spanning the first 130 amino acids followed by two repeats of ten amino acids each. It has one predicted N-glycosylation site at position 84. Furthermore, it is able to bind lactose and mannose, most likely via its lectin domain. However, the physiological function of this activity has not yet been investigated. Together with other proteins of the organic matrix, one function proposed for Perlucin is the nucleation of aragonite crystals by binding to aragonite instead of calcite. In calcium carbonate precipitation experiments, native Perlucin clearly leads to faster precipitation of CaCO3. Using atomic force microscopy it has been demonstrated that 100 mg/mL Perlucin supports nucleation on calcite in saturated CaCO3 solutions. Moreover, if Perlucin is dialysed against a saturated CaCO3 solution, the CaCO3 crystals formed incorporated Perlucin, which has recently been confirmed by Weber et al. using green fluorescent protein tagged Perlucin. Purified native Perlucin always appears very heterogeneous. This observation raised the idea that additional variants exist beside the described protein. Here we describe the cloning and expression of cDNAs encoding Perlucin from the mantle epithelia of H. laevigata. Three new splice variants of Perlucin were identified, which differ mainly in the number of a 10 amino acid repeating unit at their C-termini. Recently, it has been assumed that aragonite-associated proteins have evolved signature sequence traits of intrinsic disorder and a predicted “disordered domain” within the repeating region of a Perlucin variant described here has been identified. Interestingly, in recombinant Perlucin the number of repeats strongly influences the precipitation behaviour of CaCO3, suggesting specific physiological roles of the splice variants in the regulation of crystal growth. In addition to alternative splicing, posttranslational modifications clearly lead to heterogeneity. One motif for N-glycosylation has been identified in Perlucin. Our MALDI-ToF MS data demonstrate that the non-glycosylated protein also occurs, since the corresponding non-glycosylated peptide was identified in the peptide map of spot 4. Obviously, this does not exclude the existence of glycosylated forms of Perlucin exist. Since the glycan structures of Haliotis are unknown, the molecular masses of the corresponding glycopeptides cannot be predicted and no evidence for the non-glycosylated peptide was found in any of the other spots.

This 8 point difference is clinically meaningful, as it is larger than the difference between most categories . An additional interesting aspect of the current study was that the genetic risk score was related to the severity of depressive symptoms in two groups with wide-ranging depression scores, with a lower score related to higher depression both in healthy undergraduates and in patients with a diagnosis of depression. Future studies can examine the robustness of the genetic risk score across other conditions related to dopamine neurotransmission, where it might have utility, for example, to provide insights in the setting of Parkinson’s disease, where inter-individual response to dopaminergic therapy is highly variable. Theoretically, the dopamine genetic risk score could inform the likelihood that a drug with dopaminergic activity would be an effective antidepressant treatment choice for an individual patient. Despite its strengths, the study has a number of limitations. First, there was heterogeneity across the samples, with the two replication samples having different measures of depressive symptoms and different genes comprising the genetic risk score when compared to the discovery cohort. For example, the genetic score for STAR*D was estimated based on imputed SNPs, compared to the discovery sample, which was based on genotyped SNPs. Second, replication in the second healthy adult cohort failed to reach statistical significance, although results trended in the same direction as with the two other cohorts. This finding might reflect the fact that the 5-item POMS depression subscale used in the GSP, which measures only current mood state, lacked sufficient sensitivity to detect an association with the dopamine genetic risk score. It may also reflect the phenomenon known as the “winner’s curse”, whereby the effect of our genetic risk score could have been exaggerated in the discovery sample compared to the two replication studies. Future studies may be able to address this issue by examining more detailed measures of depressive symptoms or its specific features as well as by examining whether the genetic risk score predicts diagnoses of depression rather than depressive symptoms. Third, the direction of correlation between the DAT 9/10 polymorphism and depression is in the opposite direction of that found in a meta-analysis by Lopez-Leon. Mixed results are common with this polymorphism and the reason for the discrepancy is unclear. Fourth, dopamine effects are influenced by numerous factors such as the dynamics and concentration of its release, issues not examined in the current study. Fifth, although a great deal of evidence indicates that environmental factors may interact with genetic susceptibility to produce the final affective/behavioral phenotype, i.e, whether or not an individual will develop depression, we did not examine geneenvironment interactions in this study.