Investigating the effects of vitreous humour (crude extract) on growth and differentiation of rat mesenchymal stem cells (rMSCs) and human NTERA2 cells

It is very well documented that retinoic acid (RA) reduces growth rate by induction of cell differentiation in certain conditions and cell lines. On the other hand, hyaluronic acid (HA) is known for its growth induction on cultured cells. A natural source of HA, rabbit vitreous humour (VH), was previously shown to promote wound repair in model animals. In search for its possible mechanisms, VH extract was tested on the cultured mesenchymal stem cells and NTERA2 as human embryonal carcinoma cells in the presence of RA. Changes in some cellular and molecular markers (A2B5, Oct4, Sox2) showed that VH and possibly HA interfere with differentiating effects of RA. Therefore, this reagent may affect cell proliferation and tissue regeneration by inhibition of cell differentiation.Хорошо известно, что ретиноевая кислота (RA) снижает темпы роста, индуцируя дифференциацию клеточных линий в определенных условиях. Вместе с тем известно, что гиалуроновая кислота (HA) индуцирует рост культивируемых клеток. Ранее было показано, что естественный источник НА, стекловидное тело (VH) кролика, вызывает заживление ран у модельных животных. В поисках возможного механизма этого процесса экстракт стекловидного тела был исследован на культивируемых мезенхимальных стволовых клетках и клетках NTERA2 эмбриональной карциномы человека в присутствии RA. Изменения некоторых клеточных и молекулярных маркеров (A2B5, Oct4, Sox2) показали, что VH и, возможно, HA влияют на дифференцирующие эффекты RA. Таким образом, это вещество может влиять на пролиферацию клеток и регенерацию тканей, ингибируя дифференциацию клеток.Добре відомо, що ретиноєва кислота (RA) знижує темпи росту, індукуючи диференціацію кліткових ліній в певних умовах. Разом з тим відомо, що гіалуронова кислота (НА) індукує ріст культиво- ваних клітин. Раніше було показано, що природне джерело НА, склоподібне тіло (VH) кроля, викликає загоєння ран у модельних тварин. В пошуках можливого механізму цього процесу екстракт склоподібного тіла був досліджений на культивованих мезенхімальних стовбурових клітинах та клітинах NTERA2 ембріональної карциноми людини в присутності RA. Зміни деяких клітинних та молекулярних маркерів (A2B5, Oct4, Sox2) показали, що VH і, можливо, НА впливають на диференціюючі ефекти RA. Таким чином, ця речовина може впливати на проліферацію і регенерацію тканин, інгібуючи диференціацію клітин

Angiogenesis is induced and wound size is reduced by electrical stimulation in an acute wound healing model in human skin

Angiogenesis is critical for wound healing. Insufficient angiogenesis can result in impaired wound healing and chronic wound formation. Electrical stimulation (ES) has been shown to enhance angiogenesis. We previously showed that ES enhanced angiogenesis in acute wounds at one time point (day 14). The aim of this study was to further evaluate the role of ES in affecting angiogenesis during the acute phase of cutaneous wound healing over multiple time points. We compared the angiogenic response to wounding in 40 healthy volunteers (divided into two groups and randomised), treated with ES (post-ES) and compared them to secondary intention wound healing (control). Biopsy time points monitored were days 0, 3, 7, 10, 14. Objective non-invasive measures and H&E analysis were performed in addition to immunohistochemistry (IHC) and Western blotting (WB). Wound volume was significantly reduced on D7, 10 and 14 post-ES (p = 0.003, p = 0.002, p0.05) on day 14 in VEGF-A expression post-ES compared to controls. Furthermore, organisation of granulation tissue was improved on day 14 post-ES. This randomised controlled trial has shown that ES enhanced wound healing by reduced wound dimensions and increased VEGF-A and PLGF expression in acute cutaneous wounds, which further substantiates the role of ES in up-regulating angiogenesis as observed over multiple time points. This therapeutic approach may have potential application for clinical management of delayed and chronic wounds

An activated sulfonylating agent that undergoes general base-catalyzed hydrolysis by amines in preference to aminolysis

Activated sulfonyl derivatives, similar to acyl ones, usually undergo aminolysis with amines in water as nucleophilic attack by the amine is preferred to hydrolysis. However, despite being active sulfonyl derivatives, four-membered heterocyclic sulfonamides, β-sultams, do not undergo aminolysis in aqueous solution but preferentially react to give hydrolysis products only. The rate of the reaction of β-sultams in buffered solutions of simple primary amines shows a first-order dependence on amine concentrations attributed to general base-catalyzed hydrolysis by the amine. Even N-benzyl-4,4-dimethyl-3-oxo-β-sultam, which is both a β-sultam and a β-lactam, undergoes hydrolysis at the sulfonyl center rather than aminolysis at either the sulfonyl or acyl center. The solvent kinetic isotope effects (SKIE, kH2O/kD2O) for the amine-catalyzed hydrolyses are 1.4 and 1.9 for the hydrolysis of N-benzoyl-β-sultam and N-benzyl-4,4-dimethyl-3-oxo-β-sultam, respectively, compatible with a general base-catalyzed mechanism. The amine-catalyzed hydrolysis gives a Bronsted β value of +0.9 for both N-benzoyl β-sultam and N-benzyl-4,4-dimethyl-3-oxo-β-sultam, indicating that the general base amine is almost fully protonated in the transition state. A general base-catalyzed mechanism for hydrolysis rather than nucleophilic attack was also deduced for the reaction of N-benzyl-4,4-dimethyl-3-oxo-β-sultam with carboxylate anions based on a SKIE of 1.7−1.9 and rate constants which fit the Bronsted plot for amines. In contrast to acyl transfer reactions, those for sulfonyl transfer appear to show an inverse reactivity-selectivity relationshipthe most active compounds being the most selective. The lack of reactivity of β-sultams toward amine nucleophiles appears to be related to the mechanism of ring opening of β-sultams with a decreased reactivity toward amines relative to hydroxide ion, probably related to the expulsion of the relatively poor leaving group amide anion

Expression of seven members of the gene family encoding secretory aspartyl proteinases in Candida albicans

The opportunistic fungal pathogen Candida albicans produces secretory aspartyl proteinases, which are believed to be virulence factors in infection. We have studied the in vitro expression of seven known members of the SAP gene family in a range of strains and serotypes by Northern analysis. SAP1 and SAP3 were regulated during phenotypic switching between the white and opaque forms of the organism. The SAP2 mRNA, which was the dominant transcript in the yeast form, was found to be autoinduced by peptide products of Sap2 activity and to be repressed by amino acids. The expression of the closely related SAP4-SAP6 genes was observed only at neutral pH during serum-induced yeast to hyphal transition. No SAP7 mRNA was detected under any of the conditions or in any of the strains tested. Our data suggest that the various members of the SAP gene family may have distinct roles in the colonization and invasion of the host