Use of methanol as cryoprotectant and its effect on sox genes and proteins in chilled zebrafish embryos

Methanol is a widely used cryoprotectant (CPA) in cryopreservation of fish embryos, however little is known about its effect at the molecular level. This study investigated the effect of methanol on sox gene and protein expression in zebrafish embryos (50% epiboly) when they were chilled for 3. h and subsequently warmed and cultured to the hatching stages. Initial experiments were carried out to evaluate the chilling tolerance of 50% epiboly embryos which showed no significant differences in hatching rates for up to 6. h chilling in methanol (0.2-, 0.5- and 1. M). Subsequent experiments in embryos that had been chilled for 3. h in 1. M methanol and warmed and cultured up to the hatching stages found that sox2 and sox3 gene expression were increased significantly in hatched embryos that had been chilled compared to non-chilled controls. Sox19a gene expression also remained above control levels in the chilled embryos at all developmental stages tested. Whilst stable sox2 protein expression was observed between non-chilled controls and embryos chilled for 3. h with or without MeOH, a surge in sox19a protein expression was observed in embryos chilled for 3. h in the presence of 1. M MeOH compared to non-chilled controls and then returned to control levels by the hatching stage. The protective effect of MeOH was increased with increasing concentrations. Effect of methanol at molecular level during chilling was reported here first time which could add new parameter in selection of cryoprotectant while designing cryopreservation protocol

Evaluating the Suitability of Using Rat Models for Preclinical Efficacy and Side Effects with Inhaled Corticosteroids Nanosuspension Formulations

Inhaled corticosteroids (ICS) are often prescribed as first-line therapy for patients with asthma Despite their efficacy and improved safety profile compared with oral corticosteroids, the potential for systemic side effects continues to cause concern. In order to reduce the potential for systemic side effects, the pharmaceutical industry has begun efforts to generate new drugs with pulmonary-targeted topical efficacy. One of the major challenges of this approach is to differentiate both efficacy and side effects (pulmonary vs. systemic) in a preclinical animal model. In this study, fluticasone and ciclesonide were used as tool compounds to explore the possibility of demonstrating both efficacy and side effects in a rat model using pulmonary delivery via intratracheal (IT) instillation with nanosuspension formulations. The inhibition of neutrophil infiltration into bronchoalveolar lavage fluid (BALF) and cytokine (TNFα) production were utilized to assess pulmonary efficacy, while adrenal and thymus involution as well as plasma corticosterone suppression was measured to assess systemic side effects. Based on neutrophil infiltration and cytokine production data, the ED50s for ciclesonide and fluticasone were calculated to be 0.1 and 0.03 mg, respectively. At the ED50, the average adrenal involution was 7.6 ± 5.3% for ciclesonide versus 16.6 ± 5.1% for fluticasone, while the average thymus involution was 41.0 ± 4.3% for ciclesonide versus 59.5 ± 5.8% for fluticasone. However, the differentiation became less significant when the dose was pushed to the EDmax (0.3 mg for ciclesonide, 0.1 mg for fluticasone). Overall, the efficacy and side effect profiles of the two compounds exhibited differentiation at low to mid doses (0.03–0.1 mg ciclesonide, 0.01–0.03 mg fluticasone), while this differentiation diminished at the maximum efficacious dose (0.3 mg ciclesonide, 0.1 mg fluticasone), likely due to overdosing in this model. We conclude that the rat LPS model using IT administration of nanosuspensions of ICS is a useful tool to demonstrate pulmonary-targeted efficacy and to differentiate the side effects. However, it is only suitable at sub-maximum efficacious levels

Anesthetics Impact the Resolution of Inflammation

Local and volatile anesthetics are widely used for surgery. It is not known whether anesthetics impinge on the orchestrated events in spontaneous resolution of acute inflammation. Here we investigated whether a commonly used local anesthetic (lidocaine) and a widely used inhaled anesthetic (isoflurane) impact the active process of resolution of inflammation.Using murine peritonitis induced by zymosan and a systems approach, we report that lidocaine delayed and blocked key events in resolution of inflammation. Lidocaine inhibited both PMN apoptosis and macrophage uptake of apoptotic PMN, events that contributed to impaired PMN removal from exudates and thereby delayed the onset of resolution of acute inflammation and return to homeostasis. Lidocaine did not alter the levels of specific lipid mediators, including pro-inflammatory leukotriene B(4), prostaglandin E(2) and anti-inflammatory lipoxin A(4), in the cell-free peritoneal lavages. Addition of a lipoxin A(4) stable analog, partially rescued lidocaine-delayed resolution of inflammation. To identify protein components underlying lidocaine's actions in resolution, systematic proteomics was carried out using nanospray-liquid chromatography-tandem mass spectrometry. Lidocaine selectively up-regulated pro-inflammatory proteins including S100A8/9 and CRAMP/LL-37, and down-regulated anti-inflammatory and some pro-resolution peptides and proteins including IL-4, IL-13, TGF-â and Galectin-1. In contrast, the volatile anesthetic isoflurane promoted resolution in this system, diminishing the amplitude of PMN infiltration and shortening the resolution interval (Ri) approximately 50%. In addition, isoflurane down-regulated a panel of pro-inflammatory chemokines and cytokines, as well as proteins known to be active in cell migration and chemotaxis (i.e., CRAMP and cofilin-1). The distinct impact of lidocaine and isoflurane on selective molecules may underlie their opposite actions in resolution of inflammation, namely lidocaine delayed the onset of resolution (T(max)), while isoflurane shortened resolution interval (Ri).Taken together, both local and volatile anesthetics impact endogenous resolution program(s), altering specific resolution indices and selective cellular/molecular components in inflammation-resolution. Isoflurane enhances whereas lidocaine impairs timely resolution of acute inflammation

The Volatile Anesthetic Isoflurane Increases Endothelial Adenosine Generation via Microparticle Ecto-5′-Nucleotidase (CD73) Release

Endothelial dysfunction is common in acute and chronic organ injury. Isoflurane is a widely used halogenated volatile anesthetic during the perioperative period and protects against endothelial cell death and inflammation. In this study, we tested whether isoflurane induces endothelial ecto-5′-nucleotidase (CD73) and cytoprotective adenosine generation to protect against endothelial cell injury. Clinically relevant concentrations of isoflurane induced CD73 activity and increased adenosine generation in cultured human umbilical vein or mouse glomerular endothelial cells. Surprisingly, isoflurane-mediated induction of endothelial CD73 activity occurred within 1 hr and without synthesizing new CD73. We determined that isoflurane rapidly increased CD73 containing endothelial microparticles into the cell culture media. Indeed, microparticles isolated from isoflurane-treated endothelial cells had significantly higher CD73 activity as well as increased CD73 protein. In vivo, plasma from mice anesthetized with isoflurane had significantly higher endothelial cell-derived CD144+ CD73+ microparticles and had increased microparticle CD73 activity compared to plasma from pentobarbital-anesthetized mice. Supporting a critical role of CD73 in isoflurane-mediated endothelial protection, a selective CD73 inhibitor (APCP) prevented isoflurane-induced protection against human endothelial cell inflammation and apoptosis. In addition, isoflurane activated endothelial cells Rho kinase evidenced by myosin phosphatase target subunit-1 and myosin light chain phosphorylation. Furthermore, isoflurane-induced release of CD73 containing microparticles was significantly attenuated by a selective Rho kinase inhibitor (Y27632). Taken together, we conclude that the volatile anesthetic isoflurane causes Rho kinase-mediated release of endothelial microparticles containing preformed CD73 and increase adenosine generation to protect against endothelial apoptosis and inflammation

Токсокароз у людини: історичні, діагностичні та клінічні аспекти хвороби

The article deals with the study of the histori-cal aspects of the development of toxocarosis in humans, the algorithm for its diagnosis, clinical symptoms and treatment. Research methods: retrospective analysis of scientific and information sources on the problem of helminthic diseases. The the problem of toxocarosis has been the great social significance, helminthiasis, used for the first time an in-depth analysis of the historical, clinical and diagnostic aspects in the study of toxsocariasis in humans. The novelty of the research. Toxocarosis is a parasitic infestation, which was included in a number of pathologies, which is treated with disdain in Europe and other regions of the world in the past, but which in recent years, more and more increasing due to high incidence, high risk of infection, sustainable development, and the controversial diagnosis, as well as insufficient and incomplete therapy. Toxocarosis is a zoonotic disease nature. This means that the agents of such disease (helminthes toxocar) during the normal life cycle of living in an organism of animals, man is the intermedi-ate host and the disease can lead to serious consequences. The most common species are tt. canis and t. cati. Among these people, birds, pigs, rodents, goats, monkeys and rabbits. Man catches it when ingested eggs of t. canis or t. cati. In host larvae of paratenicheskih never ripen and remain on the second stage of development. The eggs of these worms are common environmental pollutants Wednesday for people, mostly due to the fact that many of the dogs and cats serve as pets, while many others are homeless running through the streets of most urban centers. Helminth eggs present in the feces of dogs and cats, become contagious for several weeks after their local selection Wednesday. People, especially kids, often come into contact with these eggs immediately thus accidently. Conclusions. Toxocarosis infection has depend on the number of larvae and circulating in the blood of affected organs, as well as the reaction of the immune system of the host. Hardest to recognize toxocarosis, when he is asymptomatic or symptoms is not pro-nounced. In this period can be quite long, up to several years. Infection in humans, unlike their final masters, often remains the occult. Visceral larval form (VLM) and ocular larval form (OLM) is two clinical manifestations, which lead to certain syndromes and constitute a serious health problem, wherever they occur. Keeping the patient from toxocarosis are complex and include not only different methods of diagnosis and treatment and long-term follow up. Toxocarosis is known as a single lesion is already very long, with the development of methods of diagnosis and treatment over time, allowing the gradual and profound knowledge of the helmin-thiasis of public interest to it becoming more and more intense in most of the time. The findings of the indepth study of the history and develop-ment of toxocarosis in humans, especially its algorithm of diagnosis and clinical symptoms may allow adequate technology to justify the treatment and prevention of this disease.Le but de l'étude était d'étudier en profondeur l'histoire de l'émergence et le développement de la toxocarose chez l'homme, les caractéristiques de l'algorithme pour son diagnostic, les symptômes cliniques et le traitement basé sur l'analyse du flux d'informations scientifiques. Méthodes de recherche: analyse rétrospective des sources scientifiques et d'information sur le problème des maladies helminthiques. Nouveauté scientifique: compte tenu de l'importance sociale et épidémiologique importante des helminthiases, une analyse systématique approfondie des aspects cliniques diagnostiques et historiques dans l'étude de la toxocarose chez l'homme a été utilisée pour la première fois. Conclusions Étude approfondie de l'histoire de l'émergence et le développement de la toxocarose chez l'homme, les caractéristiques de l'algorithme pour son diagnostic et les symptômes cliniques peuvent nous permettre de justifier des technologies adéquates pour le traitement et la prévention de cette maladie.Цель исследования: углубленно изучить историю возникновения и развития токсокароза у человека, особенности алгоритма его диагностики, клинической симптоматики и лечения на основании анализа научно-информационного потока информации. Методы исследования: ретроспективный анализ научно-информационных источников по проблеме гельминтозных заболеваний. Научная новизна: учитывая важное социальное, эпидемиологическое значение гельминтозов, впервые использован углубленный системный анализ исторических, диагностических клинических аспектов в изучении токсокароза у человека. Выводы. Углубленное изучение истории возникновения и развития токсокароза у человека, особенности алгоритма его диагностики и клинической симптоматики может позволить обосновать адекватные технологии лечения и профилактики данного заболевания.Мета дослідження: поглиблено вивчити історію виникнення і розвитку токсокароза у людини, особливості алгоритму його діагностики, клінічної симптоматики та лікування на підставі аналізу науково-інформаційного потоку інформації. Методи дослідження: ретроспективний аналіз науково-інформаційних джерел з проблеми гельмінтозів захворювань. Наукова новизна: враховуючи важливе соціальне, епідеміологічне значення гельмінтозів, вперше використаний поглиблений системний аналіз історія чеських, діагностичних клінічних аспектів у вивченні токсокароза у людини. Висновки. Поглиблене вивчення історії виникнення і розвитку токсокароза у людини, особливості алгоритму його діагностики та клінічної симптоматики може дозволити обгрунтувати адекватні технології лікування та профілактики даного захворювання

The effect of dalargin and hyperbaric oxygenation on the oxidant-antioxidant state of blood in patients undergone abdominal surgeries complicated with peritonitis

Досліджено вплив гіпербарооксії та даларгіну на оксидантно-антиоксидантний стан крові хворих після абдомінальних операцій, ускладнених перитонітом. Гіпербарооксія викликає підвищення в крові хворих вмісту окиснювально-модифікованих білків, активності каталази та глутатіонпероксидази. Даларгін на фоні гіпербарооксії активував антиоксидану систему крові.The effect of hyperbaric oxygen exposure and dalargin on the oxidant-antioxidant state of blood has been studied in patients who underwent abdominal surgeries complicated with peritonitis. Hyperbaric oxygenation caused an elevation of oxidatively modified proteins of blood as well as the catalase and glutathione peroxidase activities. The administration of dalargin to the patients against a background of hyperbaric oxygenation turned out to activate the blood antioxidant systems