Mango processing generates significant amounts of residues (35-65%) that may represent environmental problems owed to improper disposal. The use of mango byproducts as substrates to produce hyaluronic acid (HA) is an attractive alternative to reduce the cost of substrate. In this study, we evaluated the potential of hydrolyzates from mango peels and seeds to produce HA by Streptococcus equi. subsp. zooepidemicus. The physicochemical characterization of mango residues showed that the seeds contain a higher amount of holocellulose (cellulose and hemicellulose), which amounts 54.2% (w/w) whereas it only represents 15.5% (w/w) in the peels. Mango peels, however, are composed mainly of hot water-extractives (62% w/w, that include sucrose, fructose, glucose and organic acids). A higher concentration of monosaccharides (39.8 g/L) was obtained from the enzymatic hydrolysis (with Macerex) of peels as compared to seeds (24.8 g/L with Celuzyme). From mango peels, hydrolyzates were obtained 0.6 g/L HA, while 0.9 g/L HA were obtained with hydrolyzates from mango seeds. These results demonstrate that mango byproducts have the potential to be used for production of HA.
- MeSH
- celulosa metabolismus MeSH
- fermentace MeSH
- hydrolýza MeSH
- kyselina hyaluronová * biosyntéza metabolismus MeSH
- Mangifera * mikrobiologie chemie MeSH
- monosacharidy metabolismus MeSH
- semena rostlinná chemie mikrobiologie metabolismus MeSH
- Streptococcus equi * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
Bone morphogenetic protein-15 (BMP-15), an oocyte-derived growth factor, has been shown to play integral roles in regulation of ovarian follicular function in mammals. Despite the recognition of the physiological importance of the BMP system in regulation of gonadotropin action in the ovary, molecular mechanisms of BMP-15 effect on oocyte and somatic follicular cell functions remain poorly understood. The objective of this study was to determine the effect of BMP-15 on the FSH/LH-stimulated synthesis of hyaluronan (HA) by oocyte cumulus complexes (OCC) and progesterone by OCC and granulosa cells (GC) in the presence or absence of serum using primary porcine cultures. In addition, the effect of BMP-15 on oocyte maturation- and steroidogenesis-related transcripts after 4, 8, 16, and 24 hours of cultivation was evaluated using real-time RT-PCR. We demonstrated that the FSH/LH-induced cumulus expansion was accompanied by a significant increase in CD44, PTGS2, CYP11A1 (at 4 h) and AREG, HAS2, TNFAIP6, STAR (at 8 h) mRNAs. While FSH/LH-stimulated total HA synthesis by OCC was not affected by BMP-15 in serum-supplemented medium, its retention within the complex was significantly increased after the action of BMP-15 in comparison to FSH/LH alone (P < 0.001; 65% versus 35%, respectively). Moreover, we detected a significant increase in the expression of AREG and TNFAIP6 (both at 16 h), and CYP11A1 (at 24 h) in FSH/LH-stimulated OCC due to the action of BMP-15 compared to complexes cultured only with FSH/LH. In the presence of serum, BMP-15 markedly increased FSH/LH-stimulated progesterone secretion by OCC (about 69%) and induced a significant decrease in FSH/LH-induced progesterone release by GC (about 35%) compared to FSH/LH alone. The present results indicate that the addition of BMP-15 to the gonadotropin-stimulated OCC cultured in serum-supplemented medium might improve oocyte-cumulus maturation.
- MeSH
- gonadotropiny farmakologie MeSH
- kostní morfogenetický protein 15 farmakologie MeSH
- kultivované buňky MeSH
- kyselina hyaluronová biosyntéza MeSH
- ovariální folikul účinky léků metabolismus MeSH
- prasata MeSH
- progesteron biosyntéza MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
This review deals with molecular mechanisms controlling three important ovarian follicular processes: 1) expansion of the cumulus, 2) synthesis of the hyaluronan (HA), and 3) production of the progesterone in oocyte cumulus complexes (OCCs). The expansion of the mice cumuli induced by FSH or 8-bromo cAMP is dependent upon a specific factor(s) secreted by the oocyte (called "cumulus expansion enabling factor", CEEF). The porcine oocytes produce at least two factors that have influence on the formation and stability of the preovulatory extracellular cumulus matrix (ECM), although oocytectomy does not alter the ability of the cumulus cells to respond to FSH and forskolin by increased cAMP content, HA synthesis, and subsequent cumulus expansion, The net synthesis of HA, during the FSH-stimulated expansion of the OCCs in the presence of serum, correlates directly with accumulation of glycosaminoglycans in the ECM. In pig, insulin growth factor 1 (IGF1) is a component of the serum that promotes the FSH-stimulated synthesis and retention of HA within the expanded ECM by phosphatidylinositol 3-kinase (PI3K)/v-akt murine thymoma viral oncogene homolog (AKT)- and mitogen-activated kinase 3 and 1 (MAPK3/1)-dependent mechanisms. Mouse, porcine, bovine, and rat oocytes produce CEEF(s). Possible candidate for the CEEF in the mouse is growth differentiation factor 9 (GDF9) secreted by oocytes. In pig, GDF9 mRNA is expressed not only in the oocytes but also in the cumulus and mural granulosa cells of the growing and preovulatory follicles, although the relative abundance of the GDF9 in the somatic cells is approximately 4 times lower than in the oocytes. Cross talk between FSH/ epidermal growth factor receptor (EGFR) and transforming growth factor β (TGFβ)/GDF9 signaling pathways is essential for functional activities of the porcine OCCs, since FSH enhances EGF-induced tyrosine phosphorylation of EGFR, indicating that FSH signaling pathway may stimulate specific EGFR-regulating proteins. Also, FSH-induced synthesis of both HA and progesterone is reduced but not abolished by AG1478 (EGFR tyrosine kinase inhibitor), indicating that other signaling pathways elicited by FSH are operating in parallel. Furthermore, SMAD2/3 signaling pathway is involved in the control of both cumulus expansion and steroidogenesis in porcine OCCs, since SMAD2/3 activation by GDF9/ TGFβ produced by oocyte and/or cumulus cells, significantly affects gonadotropin-induced HA and progesterone synthesis by porcine cumulus cells. Keywords: oocyte-cumulus complex, hyaluronan, progesterone, cumulus expansion.
- MeSH
- biologické modely MeSH
- IVM techniky * metody veterinární MeSH
- krysa rodu rattus MeSH
- kumulární buňky metabolismus fyziologie MeSH
- kyselina hyaluronová biosyntéza MeSH
- myši MeSH
- oocyty metabolismus fyziologie MeSH
- prasata * metabolismus fyziologie MeSH
- proliferace buněk MeSH
- skot MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- myši MeSH
- skot MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Several lines of evidence suggest that in mice the activation of SMAD2/3 signaling by oocyte secreted factors, together with epidermal growth factor receptor (EGFR) activation, is essential to induce cumulus expansion. Here we show that inhibition of EGFR kinase in follicle stimulating hormone (FSH)-stimulated porcine oocyte-cumulus cell complex (OCCs) strongly decreases hyaluronan (HA) synthesis and its retention in the matrix, as well as progesterone synthesis. Although porcine cumulus cells undergo expansion independently of oocytes, we use biochemical and gene expression analyses to show that they do require activation of SMAD2/3 for optimal stimulation of HA synthesis and proteins involved in the organization of this polymer in the expanded matrix. Furthermore, FSH-induced progesterone synthesis by porcine cumulus cells was increased by blocking SMAD2/3 activation. In conclusion, these results support the hypothesis that an FSH-EGF autocrine loop is active in porcine OCCs, and provide the first evidence that the SMAD2/3 signaling pathway is induced by paracrine/autocrine factors in porcine cumulus cells and is involved in the control of both cumulus expansion and steroidogenesis.
- MeSH
- benzamidy farmakologie MeSH
- C-reaktivní protein metabolismus MeSH
- chinazoliny farmakologie MeSH
- dioxoly farmakologie MeSH
- epidermální růstový faktor metabolismus MeSH
- erbB receptory antagonisté a inhibitory metabolismus MeSH
- folikuly stimulující hormon metabolismus MeSH
- glukuronosyltransferasa antagonisté a inhibitory metabolismus MeSH
- isochinoliny farmakologie MeSH
- kumulární buňky metabolismus MeSH
- kyselina hyaluronová biosyntéza MeSH
- meióza účinky léků MeSH
- molekuly buněčné adheze antagonisté a inhibitory metabolismus MeSH
- myši MeSH
- oocyty enzymologie fyziologie MeSH
- prasata MeSH
- progesteron biosyntéza MeSH
- protein Smad2 antagonisté a inhibitory metabolismus MeSH
- protein Smad3 antagonisté a inhibitory metabolismus MeSH
- pyridiny farmakologie MeSH
- pyrroly farmakologie MeSH
- sérový amyloidový protein metabolismus MeSH
- signální transdukce účinky léků MeSH
- tyrphostiny farmakologie MeSH
- vývojová regulace genové exprese účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Synoviální tekutina (ST) v kloubech patří mezi významné fenomenální systémy v živé přírodě. Jedna z několika hlavních funkcí ST spočívá v adaptaci jejích viskózních vlastnosti na účinky smykových napětí. Důsledkem interakcí molekul povrchu artikulární chrupavky (ACH) s molekulami ST dochází k tokům ST ve směrech rotací a posunů bodů protilehlých povrchů ACH. ST velmi citlivě reaguje na velikosti smykových napětí a na rychlosti otáčení a posunů femorální a tibiální části kolenního kloubu při přechodech končetiny z flexe do extenze a naopak. Smyková napětí regulují v ST změny agregací makromolekul kyseliny hyaluronové. Vlivem rychlostí pootáčení a posunů povrchů ACH a vlivem rychlostí toků dochází ke změnám viskozity ST. Při valivých a posuvných pohybech femorální části kolenního kloubu a důsledkem viskozních vlastností ST dochází k rychlému snížení viskozity ST v místech největších rychlostí toků ST, tj. v rozhraní ST s povrchem ACH. Viskozita ST se směrem od protilehlých povrchů ACH směrem ke střednicové neutrální zóně (mezi povrchy ACH) zvětšuje. V neutrální zóně, vznikající v mezeře mezi artikulárními povrchy, dosahuje viskozita ST relativně větší hodnoty než u povrchů ACH.
Synovial fluid (SF) in joints belongs to significant phenomenal systems in living nature. One of several principal functions of SF consists in the adaptation of its viscose characdisteristics to the effects of shear stresses. The consequence of interactions of AC surface molecules with SF molecules are SF flows in directions of AC surface rotations. SF reacts very sensitively to the magnitude of shear stresses and to the velocity of the rotations and shifts of the femoral and tibial part of the knee joint when the lower extremity moves from flexion to extension and vice versa. Shear stresses regulate the changes of macromolecules of hyaluronic acid in SF. During the rolling movements of the femoral part of the knee joint and as a consequence of pseudoplastic properties of SF there is a fast growth in SF viscosity in places of the minimum velocities of SF, i.e. in the neutral zone. In the direction from the opposite AC surfaces towards the medial neutral zone (between AC surfaces) SF viscosity increases. In the neutral zone found in the gap between articular surfaces it reaches the higher value relatively.
- Klíčová slova
- viskozní vlastnosti,
- MeSH
- biomechanika fyziologie MeSH
- financování organizované MeSH
- kloubní chrupavka fyziologie patofyziologie patologie MeSH
- kloubní pouzdro fyziologie patofyziologie patologie MeSH
- klouby fyziologie patofyziologie patologie MeSH
- kyselina hyaluronová biosyntéza diagnostické užití MeSH
- lidé MeSH
- nemoci chrupavky etiologie patofyziologie patologie MeSH
- reologie MeSH
- statistika jako téma MeSH
- synoviální membrána fyziologie MeSH
- synoviální tekutina fyziologie MeSH
- viskozita MeSH
- Check Tag
- lidé MeSH
The purpose of the present study was to elucidate signaling pathways by which insulin like-growth factor 1 (IGF1) promotes FSH-stimulated synthesis and retention of hyaluronic acid (HA) in pig oocyte-cumulus complexes (OCCs) cultured in serum-free medium. We found that IGF1 had no effects on FSH-stimulated production of cAMP and activation of protein kinase A in the OCCs. Immunoblotting with phospho-specific antibodies showed that FSH moderately phosphorylated v-akt murine thymoma viral oncogene homolog (AKT) and mitogen-activated kinase 3 and 1 (MAPK3/1) in cumulus cells. The exposure of OCCs to both FSH and IGF1 resulted in a significant (P < 0.05) increase in AKT and MAPK3/1 phosphorylation. An inhibitor of phosphoinositide-3-kinase (PIK3), LY 294002, significantly (P < 0.05) reduced the IGF1-enhanced phosphorylation of AKT, and inhibitors of AKT (SH6) and MAPK3/1 (U0126) significantly (P < 0.05) decreased the synthesis and retention of HA stimulated by concomitant exposure of OCCs to both FSH and IGF1. The IGF1-promoted synthesis of HA was not accompanied by an increase in the relative abundance of hyaluronan synthase 2 (HAS2) mRNA in the cumulus cells. We conclude that IGF1 promotes the FSH-stimulated synthesis and retention of HA in pig OCCs by PIK3/AKT- and MAPK3/1-dependent mechanisms.
- MeSH
- AMP cyklický biosyntéza MeSH
- financování organizované MeSH
- folikulární buňky cytologie metabolismus účinky léků MeSH
- folikuly stimulující hormon farmakologie MeSH
- fosfatidylinositol-3-kinasy metabolismus MeSH
- fosforylace účinky léků MeSH
- glukuronosyltransferasa genetika metabolismus MeSH
- insulinu podobný růstový faktor I farmakologie MeSH
- kultivované buňky MeSH
- kyselina hyaluronová biosyntéza metabolismus MeSH
- mitogenem aktivovaná proteinkinasa 3 metabolismus MeSH
- oocyty metabolismus účinky léků MeSH
- prasata MeSH
- proliferace buněk účinky léků MeSH
- proteinkinasy závislé na cyklickém AMP metabolismus MeSH
- protoonkogenní proteiny c-akt metabolismus MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- MeSH
- insulinu podobný růstový faktor I farmakologie MeSH
- kyselina hyaluronová biosyntéza MeSH
- oocyty fyziologie MeSH
- ovariální folikul cytologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- přehledy MeSH
