The HGR motif is the antiangiogenic determinant of vasoinhibin : implications for a therapeutic orally active oligopeptide

The hormone prolactin acquires antiangiogenic and antivasopermeability properties after undergoing proteolytic cleavage to vasoinhibin, an endogenous prolactin fragment of 123 or more amino acids that inhibits the action of multiple proangiogenic factors. Preclinical and clinical evidence supports the therapeutic potential of vasoinhibin against angiogenesis-related diseases including diabetic retinopathy, peripartum cardiomyopathy, rheumatoid arthritis, and cancer. However, the use of vasoinhibin in the clinic has been limited by difficulties in its production. Here, we removed this barrier to using vasoinhibin as a therapeutic agent by showing that a short linear motif of just three residues (His46-Gly47-Arg48) (HGR) is the functional determinant of vasoinhibin. The HGR motif is conserved throughout evolution, its mutation led to vasoinhibin loss of function, and oligopeptides containing this sequence inhibited angiogenesis and vasopermeability with the same potency as whole vasoinhibin. Furthermore, the oral administration of an optimized cyclic retro-inverse vasoinhibin heptapeptide containing HGR inhibited melanoma tumor growth and vascularization in mice and exhibited equal or higher antiangiogenic potency than other antiangiogenic molecules currently used as anti-cancer drugs in the clinic. Finally, by unveiling the mechanism that obscures the HGR motif in prolactin, we anticipate the development of vasoinhibin-specific antibodies to solve the on-going challenge of measuring endogenous vasoinhibin levels for diagnostic and interventional purposes, the design of vasoinhibin antagonists for managing insufficient angiogenesis, and the identification of putative therapeutic proteins containing HGR.“Consejo Nacional de Ciencia y Tecnología” (CONACYT) and UNAM grant.http://link.springer.com/journal/10456ImmunologyNeurolog

On strategic choices faced by large pharmaceutical laboratories and their effect on innovation risk under fuzzy conditions

ObjectivesWe develop a fuzzy evaluation model that provides managers at different responsibility levels in pharmaceutical laboratories with a rich picture of their innovation risk as well as that of competitors. This would help them take better strategic decisions around the management of their present and future portfolio of clinical trials in an uncertain environment. Through three structured fuzzy inference systems (FISs), the model evaluates the overall innovation risk of the laboratories by capturing the financial and pipeline sides of the risk.Methods and materialsThree FISs, based on the Mamdani model, determine the level of innovation risk of large pharmaceutical laboratories according to their strategic choices. Two subsystems measure different aspects of innovation risk while the third one builds on the results of the previous two. In all of them, both the partitions of the variables and the rules of the knowledge base are agreed through an innovative 2-tuple-based method. With the aid of experts, we have embedded knowledge into the FIS and later validated the model.ResultsIn an empirical application of the proposed methodology, we evaluate a sample of 31 large pharmaceutical laboratories in the period 2008–2013. Depending on the relative weight of the two subsystems in the first layer (capturing the financial and the pipeline sides of innovation risk), we estimate the overall risk. Comparisons across laboratories are made and graphical surfaces are analyzed in order to interpret our results. We have also run regressions to better understand the implications of our results.ConclusionsThe main contribution of this work is the development of an innovative fuzzy evaluation model that is useful for analyzing the innovation risk characteristics of large pharmaceutical laboratories given their strategic choices. The methodology is valid for carrying out a systematic analysis of the potential for developing new drugs over time and in a stable manner while managing the risks involved. We provide all the necessary tools and datasets to facilitate the replication of our system, which also may be easily applied to other settings

The primary structure of acetyl-coenzyme A carboxylase messenger-RNA

Acetyl-Coenzyme A Carboxylase (ACC; acetyl-CoA:carbon dioxide ligase (ADP forming), EC 6.4.1.2), is the biotin containing enzyme that catalyzes the regulatory step in the biosynthesis of fatty acids. The primary structure of ACC mRNA, and the structural and functional features thereby deduced, are described. ACC mRNA exists as a 10 Kb, very rare mRNA species in tissues engaged in lipogenesis. Biologically active ACC mRNA has been prepared from lactating rat mammary glands. ACC cDNA clones were generated from this RNA preparation and used to obtain the primary structure of ACC mRNA. ACC mRNA contains an open reading frame 7035 nucleotides long; it encodes a polypeptide of 2345 amino acids having a M\sb{\rm r} of 265,220 dalton. The deduced protein exhibits the phosphorylation sites involved in the covalent modification of the enzyme, as well as the site for biotinylation, VMKM, which is located within the NH\sb2-terminal half of the encoded polypeptide. Sequence homologies between ACC and carbamyl phosphate synthetase, and between ACC and other biotin containing enzymes suggest that all the catalytic activities exhibited by ACC are contained in this polypeptide. This supports the hypothesis that this multifunctional enzyme has evolved through the gene fusion of separated catalytic units. Titration of the ACC gene copy number indicates that there is one copy of the ACC gene per haploid chromosome. Nonetheless, ACC mRNA exhibits a profuse 5\sp\prime end heterogeneity. Five types of ACC mRNA having different 5\sp\prime untranslated regions have been characterized. They are expressed in a tissue specific manner, and in the liver, two of them are synthesized in response to an induced lipogenic condition. Experimental evidence supports the hypothesis that the 5\sp\prime end heterogeneity is the result of alternative promoter use and differential splicing of the ACC transcriptional unit

Ventana al análisis de estados financieros

Seminario: análisis financiero para la mejor toma de decisione

Neotropical Rattlesnake (Crotalus simus) Venom Pharmacokinetics in Lymph and Blood Using an Ovine Model

The most abundant protein families in viper venoms are Snake Venom Metalloproteases (SVMPs), Snake Venom Serine Proteases (SVSPs) and Phospholipases (PLA(2)s). These are primarily responsible for the pathophysiology caused by the bite of pit-vipers; however, there are few studies that analyze the pharmacokinetics (PK) of whole venom (WV) and its protein families. We studied the pathophysiology, PK profile and differential absorption of representative toxins from venom of Neotropical Rattlesnake (Crotalus simus) in a large animal model (ovine). Toxins studied included crotoxin (the main lethal component), which causes moderate to severe neurotoxicity; SVSPs, which deplete fibrinogen; and SVMPs, which cause local tissue damage and local and systemic hemorrhage. We found that Whole Venom (WV) was highly bioavailable (86%) 60 h following intramuscular (IM) injection, and extrapolation suggests that bioavailability may be as high as 92%. PK profiles of individual toxins were consistent with their physicochemical properties and expected clinical effects. Lymph cannulated animals absorbed 1.9% of WV through lymph during the first 12 h. Crotoxin was minimally detectable in serum after intravenous (IV) injection; however, following IM injection it was detected in lymph but not in blood. This suggests that crotoxin is quickly released from the blood toward its tissue targets.Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Betaglycan (TβRIII) Is Expressed in the Thymus and Regulates T Cell Development by Protecting Thymocytes from Apoptosis

<div><p>TGF-β type III receptor (TβRIII) is a coreceptor for TGFβ family members required for high-affinity binding of these ligands to their receptors, potentiating their cellular functions. TGF-β <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044217#pone.0044217-Massague1">[1]</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044217#pone.0044217-TenDijke1">[3]</a>, bone morphogenetic proteins (BMP2/4) and inhibins regulate different checkpoints during T cell differentiation. Although TβRIII is expressed on hematopoietic cells, the role of this receptor in the immune system remains elusive. Here, we provide the first evidence that TβRIII is developmentally expressed during T cell ontogeny, and plays a crucial role in thymocyte differentiation. Blocking of endogenous TβRIII in fetal thymic organ cultures led to a delay in DN-DP transition. In addition, <em>in vitro</em> development of TβRIII^−/− thymic lobes also showed a significant reduction in absolute thymocyte numbers, which correlated with increased thymocyte apoptosis, resembling the phenotype reported in Inhibin α ^−/− thymic lobes. These data suggest that Inhibins and TβRIII may function as a molecular pair regulating T cell development.</p> </div

TβRIII deficiency results in increased apoptosis of developing thymocytes.

<p>(A) Left panel, representative CD4 versus CD8 staining dot plots from TβRIII^+/+ and TβRIII^−/− fetal thymic lobes at day 7 of culture. Histograms show the expression of active caspase 3⁺ cells in each gated thymocyte subset. Right panel, graphs represent the percentage of active caspase 3⁺ cells and the levels of expression (MFI values) in each thymocyte subset. Data are representative of three independent experiments. (B) Left panel, representative histograms show the percentage of Annexin V⁺ cells in gated thymocyte subsets. Right panel, graph shows the analysis of the percentage of Annexin V⁺ cells in thymocytes from day 7 TβRIII^+/+ or TβRIII^−/− FTOCs. Data are representative of two independent experiments. Mean values ± SEM are shown (TβRIII^+/+ n = 3 and TβRIII^−/− n = 3). Asterisks indicate statistically significant differences (** p≤0.05).</p

The blocking of TβRIII in FTOCs alters T cell development.

<p>E14 thymic lobes were cultured in the presence of anti-TβRIII antibody or in the presence of pre-immune serum (control lobe). At day 3 and 7 of culture thymic lobes were disaggregated, counted and stained with antibodies to CD4, CD8. (A) Representative CD4 versus CD8 staining dot plots. (B). Comparative graphs represent the percentages of DN, DP, CD4SP and CD8SP thymocytes obtained after 3 and 7 days of culture between both treatments. (C) Analysis of cell numbers in non-treated and anti-TβRIII treated FTOCs at day 3 and 7. Data are representative of two independent experiments. Mean values ± SEM are shown (n  = 7 per group for day 3, and n  = 9 per group for day 7). Asterisks indicate *p≤0.05.</p