The Chemical State of the Calcium Reacting in the Coagulation of Blood

1. The widely accepted theory that calcium participates in the coagulation mechanism in the form of Ca++ and acts as a catalyst is not in accord with several important experimental findings: (a) The anticoagulant action of sodium oxalate is much slower than the precipitation of ionized calcium as the oxalate salt. (b) Sodium citrate begins to depress prothrombin activity at a concentration at which ionized calcium is still present. The inability of tricalcium phosphate to adsorb prothrombin from citrated plasma indicates that citrate forms a complex with prothrombin and it is postulated that prothrombin is thereby inactivated. (c) In plasma which is decalcified, i.e. in which the Ca++ is markedly reduced, the labile factor of prothrombin rapidly decreases. A concentration of 0.01 M sodium citrate sufficient to inhibit coagulation does not depress Ca++ enough to cause diminution of the labile factor, whereas when the concentration is increased to 0.02 M the labile factor decreases as rapidly as in oxalated plasma. 2. It is postulated that calcium functions in coagulation not as Ca++ but as combined with a component which is part of the prothrombin complex that is not adsorbed by tricalcium phosphate. A concentration of sodium citrate just sufficient to inhibit coagulation is not enough to remove calcium from the essential prothrombin component. The primary anticoagulant action of sodium citrate is therefore not decalcification but antiprothrombic. 3. It has been shown that citrated plasma is basically different from oxalated plasma in several important aspects. Unless cognizance is taken of these differences, serious errors and misinterpretations of experimental findings may be made

Clindamycin as trigger of uroporphyrinuria

Administration of clindamycin for the management of a staphylococcal abscess of the abdominal wall was followed by development of a non-pruriginous, macular, blistery rash involving the areas of the body exposed to sunlight.  This was accompanied by significant elevation of urinary uroporphyrin possibly related to the ability of clindamycin to inhibit the enzyme CYP3A4 of the P450 system

Identification of side population cells in mouse primordial germ cells and prenatal testis

In mammals, the stem cells of spermatogenesis are derived from an embryonic cell population called primordial germ cells (PGCs). Spermatogonial stem cells displaying the "side population" (SP) phenotype have been identified in the immature and adult mouse testis, but noting is known about the expression of the SP phenotype during prenatal development of germ cells. The SP phenotype, defined as the ability of cells to efflux fluorescent dyes such as Hoechst, is common to several stem/progenitor cell types. In the present study, we analyzed and characterized the Hoechst SP via cytofluorimetric analysis of disaggregated gonads at different time points during embryonic development in mice. To directly test the hypothesis that the SP phenotype is a feature of germ cell lineage, experiments were performed on transgenic animals expressing enhanced green fluorescent protein (EGFP) under the control of the Oct4 promoter, to identify early germ cells up to PGCs. We found that prenatal gonads contain a fraction of SP cells at each stage analyzed, and the percentage of cells in the SP fraction decreases as development proceeds. Surprisingly, more than 50% of the PGCs displayed the SP phenotype at 11.5 dpc (days post coitum). The percentage of germ cells with the SP phenotype decreased steadily with development, to less than 1% at 18.5 dpc. Cytofluorimetric analysis along with immunocytochemistry performed on sorted cells indicated that the SP fraction of prenatal gonads, as in the adult testis, was heterogeneous, being composed of both somatic and germ cells. Both cell types expressed the ABC transporters Abcg2, Abcb1a, Abcb1b and Abcc1. These findings provide evidence that the SP phenotype is a common feature of PGCs and identifies a subpopulation of fetal testis cells including prospermatogonia whose differentiation fate remains to be investigated. © 2011 UBC Press

Signal transduction in the Sertoli cell: serum modulation of the response to FSH

Immature Sertoli cells of the testicular seminiferous tubule maintain the expression of their differentiated phenotype when cultured in unsupplemented medium. In preliminary experiments we observed that foetal bovine serum (FBS) stimulates polyphosphoinositides (PI) hydrolysis in Sertoli cells. We then evaluated the effect of serum on the function of the immature Sertoli cell in culture, in terms of cAMP and estrogen production. Treatment of Sertoli cells for 30 min with 1–10% FBS had no effect on basal cAMP accumulation but abolished the response to FSH. The serum concentration producing half-maximal inhibition of the FSH-dependent cAMP accumulation was 0.5–1%. Comparison of the FSH-dose-response in the absence or presence of serum showed a decreased maximal response when serum was present. Sertoli cells exposed to serum were also less responsive to the β-adrenergic agonist isoproterenol, to cholera toxin, and to forskolin. The serum inhibition was rapidly reversed upon removal of serum or incubating the cells with the phosphodiesterase inhibitor MIX (methyl-isobutyl-xanthine). Similarly to what observed with cAMP, serum affected androgen aromatization stimulated by FSH, isoproterenol, cholera toxin, forskolin and dibutyryl cAMP. These data indicate that factors present in serum can act as modulators of the Sertoli cell function in vitro by rapidly and reversibly inhibiting the cAMP and steroidogenic response of the Sertoli cell to FSH

Activation of Jun N-terminal Kinase/Stress-activated Protein Kinase Pathway by Tumor Necrosis Factor α Leads to Intercellular Adhesion Molecule-1 Expression

Tumor necrosis factor alpha (TNF-alpha) is a cytokine implicated in the pathogenesis of numerous chronic and acute inflammatory conditions. We have previously shown that mouse Sertoli cells respond to TNF-alpha by increasing interleukin-6 production and intercellular adhesion molecule-1 (ICAM-1) expression (1). In this cell type TNF-alpha activates the mitogen-activated protein kinase (MAPK) pathways p42/p44 MAPK, JNK/SAPK, and p38, the last of which is responsible for interleukin-6 production (1). To determine which MAPK signaling pathway is required for TNF-alpha induction of ICAM-1 expression, we have utilized the protein kinase inhibitor dimethylaminopurine, demonstrating that treatment of Sertoli cells with such compound significantly reduced ICAM-1 expression and JNK/SAPK activation. Moreover, dimethylaminopurine treatment increased the expression of MAPK phosphatase-2, providing a possible mechanism of action of this compound. By using agonist antibodies to p55 and to p75 TNF-alpha receptors and both human and mouse TNF-alpha, we demonstrate that both TNF receptors are expressed and that only the p55 receptor is involved in ICAM-1 expression. The p55 receptor activates all of the three pathways, whereas p75 failed to activate any of the MAPKs. Altogether our results demonstrate that TNF-alpha up-regulates ICAM-1 expression through the activation of the JNK/SAPK transduction pathway mediated by the p55 receptor

Fortalecimiento de redes socioproductivas como base de la transformación social

Ponencia presentada en el I Congreso de Extensión de la AUGM. Montevideo, Uruguay, 6 al 8 de noviembre de 2013El presente trabajo, fue realizado dentro del marco del Programa de Voluntariado Universitario y a partir del trabajo interinstitucional entre la Facultad de Ciencias Agropecuarias de la Universidad Nacional de Córdoba (FCA) y el Pro Huerta del INTA (PH), en pos de contribuir a co-construir un tejido productivo, social y político que garantizara un proceso de transformación de la sociedad basado en la ética de la equidad y la participación democrática.Fil: Barrientos, Mario Alberto. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; Argentina.Fil: Stefanini, Ximena Verónica. Instituto Nacional de Tecnología Agropecuaria. Agencia de Extensión Rural Córdoba. Pro Huerta; Argentina

Spermatogonial kinetics in humans

The human spermatogonial compartment is essential for daily production of millions of sperm. Despite this crucial role, the molecular signature, kinetic behavior and regulation of human spermatogonia are poorly understood. Using human testis biopsies with normal spermatogenesis and by studying marker protein expression, we have identified for the first time different subpopulations of spermatogonia. MAGE-A4marks all spermatogonia, KITmarks all Bspermatogonia and UCLH1 all Apale-dark (Ap-d) spermatogonia. We suggest that at the start of the spermatogenic lineage there are Ap-d spermatogonia that are GFRA1High, likely including the spermatogonial stem cells. Next, UTF1 becomes expressed, cells become quiescent and GFRA1 expression decreases. Finally, GFRA1 expression is lost and subsequently cells differentiate into B spermatogonia, losing UTF1 and acquiring KIT expression. Strikingly, most human Ap-d spermatogonia are out of the cell cycle and even differentiating type B spermatogonial proliferation is restricted. A novel scheme for human spermatogonial development is proposed that will facilitate further research in this field, the understanding of cases of infertility and the development of methods to increase sperm output

Tympanic Membrane Collagen Expression by Dynamically Cultured Human Mesenchymal Stromal Cell/Star-Branched Poly(ε-Caprolactone) Nonwoven Constructs

The tympanic membrane (TM) primes the sound transmission mechanism due to special fibrous layers mainly of collagens II, III, and IV as a product of TM fibroblasts, while type I is less represented. In this study, human mesenchymal stromal cells (hMSCs) were cultured on star-branched poly("-caprolactone) (*PCL)-based nonwovens using a TM bioreactor and proper dierentiating factors to induce the expression of the TM collagen types. The cell cultures were carried out for one week under static and dynamic conditions. Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) were used to assess collagen expression. A Finite Element Model was applied to calculate the stress distribution on the scaolds under dynamic culture. Nanohydroxyapatite (HA) was used as a filler to change density and tensile strength of *PCL scaolds. In dynamically cultured *PCL constructs, fibroblast surface marker was overexpressed, and collagen type II was revealed via IHC. Collagen types I, III and IV were also detected. Von Mises stress maps showed that during the bioreactor motion, the maximum stress in *PCL was double that in HA/*PCL scaolds. By using a *PCL nonwoven scaold, with suitable physico-mechanical properties, an oscillatory culture, and proper dierentiative factors, hMSCs were committed into fibroblast lineage-producing TM-like collagens

Piezoelectric energy harvesting solutions

This paper reviews the state of the art in piezoelectric energy harvesting. It presents the basics of piezoelectricity and discusses materials choice. The work places emphasis on material operating modes and device configurations, from resonant to non-resonant devices and also to rotational solutions. The reviewed literature is compared based on power density and bandwidth. Lastly, the question of power conversion is addressed by reviewing various circuit solutions

Consensus Report of Working Group 2:Soft Tissue Management

OBJECTIVES: The task of working Group 2 at the 6th Consensus Meeting of the European Association for Osseointegration was to comprehensively assess the effects of soft tissue augmentation procedures at dental implant sites on clinical, radiographic and patient‐reported outcome measures (PROMs) including an overview on available outcome measures and methods of assessment. MATERIALS AND METHODS: Three systematic reviews and one critical review were performed in advance on (i) the effects of soft tissue augmentation procedures on clinical, radiographic and aesthetic outcomes, (ii) reliability and validity of outcome measures and methods of assessment and (iii) PROMs applied in clinical studies for soft tissue augmentation procedures at dental implant sites. Major findings, consensus statements, clinical recommendations and implications for future research were discussed in the group and approved during the plenary sessions. RESULTS: The four reviews predominantly revealed: Soft tissue augmentation procedures in conjunction with immediate and delayed implant placement result in superior aesthetic outcomes compared to no soft tissue augmentation in the zone of aesthetic priority. Soft tissue augmentation procedures have a limited effect on marginal bone level changes compared to implant sites without soft tissue augmentation. Clinically relevant parameters (gingival index, mucosal recession) and plaque control improve at implant sites when the width of keratinised mucosa is increased. A variety of aesthetic indices have been described with good reliability. Pink Esthetic Score and Complex Esthetic Index are the most validated aesthetic indices for single implants, though. Superimposed digital surface scans are most accurate to assess profilometric tissue changes. PROMs following soft tissue augmentation procedures have been assessed using various forms of questionnaires. Soft tissue augmentation had a limited effect on PROMs. CONCLUSIONS: Soft tissue augmentation procedures are widely applied in conjunction with implant therapy. Depending on the indication of these interventions, clinical, radiographic and aesthetic outcomes may improve, whereas the effect on PROMs is limited