Mitochondria-Targeted Antioxidant SkQ1 Improves Dermal Wound Healing in Genetically Diabetic Mice

Oxidative stress is widely recognized as an important factor in the delayed wound healing in diabetes. However, the role of mitochondrial reactive oxygen species in this process is unknown. It was assumed that mitochondrial reactive oxygen species are involved in many wound-healing processes in both diabetic humans and animals. We have applied the mitochondria-targeted antioxidant 10-(6′-plastoquinonyl)decyltriphenylphosphonium (SkQ1) to explore the role of mitochondrial reactive oxygen species in the wound healing of genetically diabetic mice. Healing of full-thickness excisional dermal wounds in diabetic C57BL/KsJ-db−/db− mice was significantly enhanced after long-term (12 weeks) administration of SkQ1. SkQ1 accelerated wound closure and stimulated epithelization, granulation tissue formation, and vascularization. On the 7th day after wounding, SkQ1 treatment increased the number of α-smooth muscle actin-positive cells (myofibroblasts), reduced the number of neutrophils, and increased macrophage infiltration. SkQ1 lowered lipid peroxidation level but did not change the level of the circulatory IL-6 and TNF. SkQ1 pretreatment also stimulated cell migration in a scratch-wound assay in vitro under hyperglycemic condition. Thus, a mitochondria-targeted antioxidant normalized both inflammatory and regenerative phases of wound healing in diabetic mice. Our results pointed to nearly all the major steps of wound healing as the target of excessive mitochondrial reactive oxygen species production in type II diabetes

Novel Biodegradable Polymeric Microparticles Facilitate Scarless Wound Healing by Promoting Re-epithelialization and Inhibiting Fibrosis

Despite decades of research, the goal of achieving scarless wound healing remains elusive. One of the approaches, treatment with polymeric microcarriers, was shown to promote tissue regeneration in various in vitro models of wound healing. The in vivo effects of such an approach are attributed to transferred cells with polymeric microparticles functioning merely as inert scaffolds. We aimed to establish a bioactive biopolymer carrier that would promote would healing and inhibit scar formation in the murine model of deep skin wounds. Here we characterize two candidate types of microparticles based on fibroin/gelatin or spidroin and show that both types increase re-epithelialization rate and inhibit scar formation during skin wound healing. Interestingly, the effects of these microparticles on inflammatory gene expression and cytokine production by macrophages, fibroblasts, and keratinocytes are distinct. Both types of microparticles, as well as their soluble derivatives, fibroin and spidroin, significantly reduced the expression of profibrotic factors Fgf2 and Ctgf in mouse embryonic fibroblasts. However, only fibroin/gelatin microparticles induced transient inflammatory gene expression and cytokine production leading to an influx of inflammatory Ly6C+ myeloid cells to the injection site. The ability of microparticle carriers of equal proregenerative potential to induce inflammatory response may allow their subsequent adaptation to treatment of wounds with different bioburden and fibrotic content

<i>Komagataella phaffii</i> as a Platform for Heterologous Expression of Enzymes Used for Industry

In the 1980s, Escherichia coli was the preferred host for heterologous protein expression owing to its capacity for rapid growth in complex media; well-studied genetics; rapid and direct transformation with foreign DNA; and easily scalable fermentation. Despite the relative ease of use of E. coli for achieving the high expression of many recombinant proteins, for some proteins, e.g., membrane proteins or proteins of eukaryotic origin, this approach can be rather ineffective. Another microorganism long-used and popular as an expression system is baker’s yeast, Saccharomyces cerevisiae. In spite of a number of obvious advantages of these yeasts as host cells, there are some limitations on their use as expression systems, for example, inefficient secretion, misfolding, hyperglycosylation, and aberrant proteolytic processing of proteins. Over the past decade, nontraditional yeast species have been adapted to the role of alternative hosts for the production of recombinant proteins, e.g., Komagataella phaffii, Yarrowia lipolytica, and Schizosaccharomyces pombe. These yeast species’ several physiological characteristics (that are different from those of S. cerevisiae), such as faster growth on cheap carbon sources and higher secretion capacity, make them practical alternative hosts for biotechnological purposes. Currently, the K. phaffii-based expression system is one of the most popular for the production of heterologous proteins. Along with the low secretion of endogenous proteins, K. phaffii efficiently produces and secretes heterologous proteins in high yields, thereby reducing the cost of purifying the latter. This review will discuss practical approaches and technological solutions for the efficient expression of recombinant proteins in K. phaffii, mainly based on the example of enzymes used for the feed industry

Mitochondria-Targeted Antioxidant SkQ1 Prevents the Development of Experimental Colitis in Mice and Impairment of the Barrier Function of the Intestinal Epithelium

Mitochondria-targeted antioxidants have become promising candidates for the therapy of various pathologies. The mitochondria-targeted antioxidant SkQ1, which is a derivative of plastoquinone, has been successfully used in preclinical studies for the treatment of cardiovascular and renal diseases, and has demonstrated anti-inflammatory activity in a number of inflammatory disease models. The present work aimed to investigate the therapeutic potential of SkQ1 and C12TPP, the analog of SkQ1 lacking the antioxidant quinone moiety, in the prevention of sodium dextran sulfate (DSS) experimental colitis and impairment of the barrier function of the intestinal epithelium in mice. DSS-treated animals exhibited weight loss, bloody stool, dysfunction of the intestinal epithelium barrier (which was observed using FITC-dextran permeability), reduced colon length, and histopathological changes in the colon mucosa. SkQ1 prevented the development of clinical and histological changes in DSS-treated mice. SkQ1 also reduced mRNA expression of pro-inflammatory molecules TNF, IL-6, IL-1&beta;, and ICAM-1 in the proximal colon compared with DSS-treated animals. SkQ1 prevented DSS-induced tight junction disassembly in Caco-2 cells. Pretreatment of mice by C12TPP did not protect against DSS-induced colitis. Furthermore, C12TPP did not prevent DSS-induced tight junction disassembly in Caco-2 cells. Our results suggest that SkQ1 may be a promising therapeutic agent for the treatment of inflammatory bowel diseases, in particular ulcerative colitis

Biological Studies of New Implant Materials Based on Carbon and Polymer Carriers with Film Heterostructures Containing Noble Metals

This paper presents pioneering results on the evaluation of noble metal film hetero-structures to improve some functional characteristics of carbon-based implant materials: carbon-composite material (CCM) and carbon-fiber-reinforced polyetheretherketone (CFR-PEEK). Metal-organic chemical vapor deposition (MOCVD) was successfully applied to the deposition of Ir, Pt, and PtIr films on these carriers. A noble metal layer as thin as 1 µm provided clear X-ray imaging of 1–2.5 mm thick CFR-PEEK samples. The coated and pristine CCM and CFR-PEEK samples were further surface-modified with Au and Ag nanoparticles (NPs) through MOCVD and physical vapor deposition (PVD) processes, respectively. The composition and microstructural features, the NPs sizes, and surface concentrations were determined. In vitro biological studies included tests for cytotoxicity and antibacterial properties. A series of samples were selected for subcutaneous implantation in rats (up to 3 months) and histological studies. The bimetallic PtIr-based heterostructures showed no cytotoxicity in vitro, but were less biocompatible due to a dense two-layered fibrous capsule. AuNP heterostructures on CFR-PEEK promoted cell proliferation in vitro and exhibited a strong inhibition of bacterial growth (p < 0.05) and high in vitro biocompatibility, especially Au/Ir structures. AgNP heterostructures showed a more pronounced antibacterial effect, while their in vivo biocompatibility was better than that of the pristine CFR-PEEK, but worse than that of AuNP heterostructures

The Issue of Obedience in the Anglican Confessional Thought in the mid 1629s

Тема послушания стала актуальна для англиканской конфессио- нальной мысли после Реформации. Эта проблематика вновь начала обсуждаться в 1620-х годах, когда королевская власть попыталась собрать не легитимный, по мнению многих подданных, принудительный заем. В 1627 г. изрядное количество дворян отказалось раскошеливаться и угодило в тюрьму. Следствием стало из- вестное дело «пяти рыцарей», дебаты в парламенте 1628 г. и принятая палатами знаменитая «Петиция о праве». В конце сентября 1626 г. Карл I отправил письмо епископам, призывая их обратиться с церковных кафедр к прихожанам с просьбой «помочь и поддержать» личными средствами «защиту королевства». Набирающий политический вес, лидер арминианского крыла в церкви епископ Бата и Уэльса Уильям Лод адресовал духовенству инструкцию, призывающую «напомнить под- данным о долге послушания». В статье разбираются четыре проповеди: первая именовалась «Апостольское послушание» и была произнесена 22 февраля 1627 г. в Нортгемптоне Робертом Сибторпом, вторая, названная в печати просто «Про- поведь», принадлежит капеллану короля, настоятелю Кентерберийского собора Исааку Баргрейву и была произнесена в присутствии монарха 27 марта 1627 г., еще две под общим названием «Религия и верность» прозвучали 4 июля 1627 г. перед Карлом I в королевском дворце в Отланде, графство Сари, и 29 июля в Олдертоне, графство Сэффолк из уст другого королевского капеллана Роджера Мэнверинга. Все проповеди для большего эффекта были по повелению короля опубликованы. Англиканский клир интересовался проблемой послушания и продвигал ее среди прихожан в интерпретации, угодной королевской власти, а именно: как важнейший духовный принцип, на котором держится установленный порядок. Англиканское духовенство считало монарха божьим наместником на земле, а неповиновение королю склонно было расценивать как сопротивление самому Господу, как по- пытку разрушить устроенное им иерархическое системное единство. Проповеди показывают, что англиканские клирики были близки к тому, чтобы поставить повиновение монарху выше повиновения закону и традиции. Некоторые из них полагали, что миссия поддерживать порядок дает королю право по своему усмо- трению назначать и собирать денежные субсидии с подданных. Однако были и те, кто расценивал такие заявления, как недопустимую попытку расширить границы королевской прерогативы и как покушения на исконные вольности . The idea of obedience emerged full blown for the Anglican confessional thought after the Reformation. This problem came under discussion again in 1620s, when the royal power tried to collect non-legitimate in the eyes of many subjects forced loans. In 1627 a number of noblemen refused to dip into their purse and were imprisoned. It resulted in famous Five Knights’ case, parliament debates in 1628, which contributed to the enactment of the Petition of Right. In late September 1626 Charles I sent a letter to his bishops, demanding to appeal from their cathedrae to the parish with a request to “help and support protection of the royalty” by their personal means. Coming into political force leader of the Arminian church wing bishop of Bath and Wells William Laud addressed an instruction to the clergy, which demanded to “remind subjects about their debt of obedience”. The present paper studies four sermons: the first was called Apostolic Obedience and was delivered by Robert Sibthorpe on the 22nd of February, 1627 in Northampton, the second, simply called in press The Sermon belonged to the king’s chaplain, Dean of Canterbury Isaac Bargrave and was delivered before the king’ majesty on the 27th of March, 1627. Two sermons under common name “Religion and Allegiance” were preached before Charles I by the king’s chaplain Roger Maynwaring the one on the 4th of July 1627 at the king’s palace at Oatlands Surrey, the other on the 29th of July at Alderton Suffolk. To heighten an effect all sermons were published by order of his majesty. The Anglican clergy was interested in the problem of obedience and promoted it among the parish in interpretation acceptable before the royal power, namely as the most impotent religious principle, lying in the ground of the stated order. The Anglican Church considered the king to be the vicar of God on earth, thus disobedience to the king was judged as rebellion to Christ Himself, i.e. an attempt to destroy a hierarchically systemized community settled by Him. The sermons show that the Anglican clergy was close to place obedience to the monarch higher than obedience to law or tradition. Some of them believed that a mission to support the order gave the king a right to set and collect monetary contributions from his subjects to his own discretion. Nonetheless, the others regarded such claims as an intolerable attempt to broaden the limits of royal prerogative and as aggression upon ancient liberties of subjects