Tissue Balance Research Articles

Cold stress-induced autophagy and apoptosis disorders are mainly mediated by AMPK/PPAR/PI3K/AKT/mTOR pathways

Under cold stress, the processes of autophagy, apoptosis and energy metabolism are pivotal for sustaining energy and tissue balance. However, the molecular regulatory mechanisms and interactions underlying these processes are still largely unknown. In this study, the molecular mechanisms underlying the regulation of energy homeostasis and tissue homeostasis of Cranoglanis bouderius (13 ± 0.5 g) under cold stress conditions were investigated using histological, metabolomic, and transcriptomic approaches after programmed cooling (25 °C, 20 °C, and 15 °C). Results revealed an uptick in the serum antioxidant activity of C. bouderius during cold stress, contrasted with a decrease in cholesterol levels. Histological assessments revealed extensive damage, marked by evident apoptotic signals, in the gills, liver, and muscles. Transmission electron microscopy and mitochondrial membrane potential analysis also indicated that cold stress induced mitochondrial damage in the liver of C. bouderius, resulting in decreased mitochondrial membrane potential and increased mitochondrial fusion and autophagy. The integrated analysis of metabolomics and transcriptomics suggests that under cold stress, energy homeostasis is primarily regulated by the coordinated action of the AMPK and PPAR signaling pathways, with potential modulation by metabolites like epinephrine. The obstruction of biosynthetic processes is primarily associated with the inhibition of signaling pathways such as PI3K/AKT/mTOR and protein processing in the endoplasmic reticulum. Sustained activation of AMPK alleviates the PI3K/AKT/mTOR-mediated inhibition of autophagy and apoptosis, potentially underpinning the transition from autophagy-mediated apoptosis suppression to apoptosis promotion during cold stress. In summary, these findings elucidate the consequences arising from the altered interactions within the AMPK/PPAR/PI3K/AKT/mTOR pathway under cold stress conditions and offer a theoretical basis for enhancing the ability of fish species to cope with cold stress.

Tissue inhibitors of metalloproteinases are proteolytic targets of matrix metalloproteinase 9

Extracellular proteolysis and turnover are core processes of tissue homeostasis. The predominant matrix-degrading enzymes are members of the Matrix Metalloproteinase (MMP) family. MMPs extensively degrade core matrix components in addition to processing a range of other factors in the extracellular, plasma membrane, and intracellular compartments. The proteolytic activity of MMPs is modulated by the Tissue Inhibitors of Metalloproteinases (TIMPs), a family of four multi-functional matrisome proteins with extensively characterized MMP inhibitory functions. Thus, a well-regulated balance between MMP activity and TIMP levels has been described as critical for healthy tissue homeostasis, and this balance can be chronically disturbed in pathological processes. The relationship between MMPs and TIMPs is complex and lacks the constraints of a typical enzyme-inhibitor relationship due to secondary interactions between various MMPs (specifically gelatinases) and TIMP family members. We illustrate a new complexity in this system by describing how MMP9 can cleave members of the TIMP family when in molar excess. Proteolytic processing of TIMPs can generate functionally altered peptides with potentially novel attributes. We demonstrate here that all TIMPs are cleaved at their C-terminal tails by a molar excess of MMP9. This processing removes the N-glycosylation site for TIMP3 and prevents the TIMP2 interaction with latent proMMP2, a prerequisite for cell surface MMP14-mediated activation of proMMP2. TIMP2/4 are further cleaved producing ∼14 kDa N-terminal proteins linked to a smaller C-terminal domain through residual disulfide bridges. These cleaved TIMP2/4 complexes show perturbed MMP inhibitory activity, illustrating that MMP9 may bear a particularly prominent influence upon the TIMP:MMP balance in tissues.

Assessment of oxygen extraction rate changes following red blood cell transfusion in the intensive care unit: a protocol for a prospective observational non-interventional study

IntroductionHaemoglobin transfusion thresholds have been used in the intensive care unit (ICU) to guide red blood cell transfusion (RBCT) decisions. Recent research has also focused on physiological indicators of tissue...

P2X7 receptor antagonist A-438079 alleviates oxidative stress of lung in LPS-induced septic rats.

Sepsis is a deadly systemic inflammatory response of the body against infection resulting in immune response, cell differentiation and organ damage. Endotoxemia is one of the causes of sepsis-related acute respiratory distress and respiratory burst is an important generator of oxidants. Inflammation may be aggravated by overexpression of ATP-gated purinergic receptors (i.e., P2X7R) following cell damage. We aimed to evaluate the effects of P2X7R antagonist A-438079 on lung oxidative status and the receptor expression in endotoxemia of sepsis. Rats were subjected to sepsis by E. coli lipopolysaccharide (LPS) and treated with 15mg/kg A-438079. The increase in circulatory IL-1β and IL-8 concentrations in LPS group confirmed the systemic inflammatory response to endotoxemia compared with Control groups (p < 0.001). Besides, there was an increase in P2X7R expression in lung tissue after LPS administration. Compared with Control groups, there were significant increases in the values of malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) (p < 0.001), and myeloperoxidase (MPO) (p < 0.05) in lung tissue of LPS group. P2X7R expression in lung and IL-1β level in blood did not increase in LPS + A-438079 group. A-438079 decreased the lung levels of MDA, GSH, CAT and SOD (p < 0.001), and MPO (p < 0.01) in septic rats. As a result, administration of pathogen-associated LPS led to increased P2X7R expression into lung tissue and elevated lipid peroxidation product MDA with regard to oxidative damage. The P2X7R antagonist A-438079 alleviated the oxidative stress of lung with a balance of tissue oxidant/antioxidant factors in experimental sepsis in rats.

RESVERATROL PROTECTS WHOLE BODY HEAT STRESS-INDUCED TESTICULAR DAMAGE IN RAT MODEL

Objective: The local thermoregulation in testis is important for optimum spermatozoa development. Excessive heat hampers this regulation resulting in alteration of normal testicular function. The present investigation confirms the role of free radicals in hyperthermia induced oxidative damage in testis and elucidates the dose-dependent ameliorating effect of resveratrol (RSV) against testicular oxidative damage. The aim of the present investigation is also to observe the role of selective concentration of RSV on heat induced oxidative changes in the damaged tissue. Methods: 48 male Wister rats were exposed to hyperthermic condition for the past 7 days of the total 21 days of experiment. RSV was pre- and co-treated with heat stress daily in a dose-dependent manner (1 mg, 5 mg, and 10 mg/kg body weight) for 21days. Results: Reactive oxygen species level was estimated using flow cytometry. Enhancement of hepatotoxicity markers in serum, lipid peroxidation and decreasing antioxidant status in the testis homogenate demonstrated that the oxidative damage in heat exposed tissue. Conclusion: Histological study along with biochemical and molecular assessment of the redox balance of testicular tissue in the present study revealed that RSV significantly ameliorated the heat induced damage in testis. The findings suggest that RSV is an effective antioxidant polyphenolic compound that can protect testis against hyperthermia induced oxidative damage.

Mathematical Modeling of Lymph Node Drainage Function by Neural Network

The lymph node (LN) represents a key structural component of the lymphatic system network responsible for the fluid balance in tissues and the immune system functioning. Playing an important role in providing the immune defense of the host organism, LNs can also contribute to the progression of pathological processes, e.g., the spreading of cancer cells. To gain a deeper understanding of the transport function of LNs, experimental approaches are used. Mathematical modeling of the fluid transport through the LN represents a complementary tool for studying the LN functioning under broadly varying physiological conditions. We developed an artificial neural network (NN) model to describe the lymph node drainage function. The NN model predicts the flow characteristics through the LN, including the exchange with the blood vascular systems in relation to the boundary and lymphodynamic conditions, such as the afferent lymph flow, Darcy’s law constants and Starling’s equation parameters. The model is formulated as a feedforward NN with one hidden layer. The NN complements the computational physics-based model of a stationary fluid flow through the LN and the fluid transport across the blood vessel system of the LN. The physical model is specified as a system of boundary integral equations (IEs) equivalent to the original partial differential equations (PDEs; Darcy’s Law and Starling’s equation) formulations. The IE model has been used to generate the training dataset for identifying the NN model architecture and parameters. The computation of the output LN drainage function characteristics (the fluid flow parameters and the exchange with blood) with the trained NN model required about 1000-fold less central processing unit (CPU) time than computationally tracing the flow characteristics of interest with the physics-based IE model. The use of the presented computational models will allow for a more realistic description and prediction of the immune cell circulation, cytokine distribution and drug pharmacokinetics in humans under various health and disease states as well as assisting in the development of artificial LN-on-a-chip technologies.

Total Knee Arthroplasty Conversion after Open-wedge High Tibial Osteotomy

Background: Open-wedge High Tibial Osteotomy (HTO) is an effective treatment for Osteoarthritis (OA) of the knee; however, postoperative changes due to HTO can make subsequent Total Knee Arthroplasty (TKA) technically difficult and can affect long-term survival. Case Report: Medio-lateral stability along with balanced flexion and extension gaps provides a good functional outcome. A 71-year-old man underwent TKA conversion after open-wedge HTO 12 years earlier; the conversion was performed due to arthritic deterioration in the knee. Medial instability was noted because of poor ligament balancing. The residual medial instability was managed with a knee brace due to the unavailability of the constrained prosthesis. Management of persistent mediolateral instability persisting after soft tissue release and balancing ideally requires varus-valgus constraint. Conclusion: TKA conversion after open-wedge HTO requires a special technique and careful preparation before surgery.

The Assessment of Fixed Orthodontic Treatment Successfulness Using American Board of Orthodontics System in Airlangga University Dental Hospital

Background: The purpose of orthodontic treatment is to improve efficient function, tissue balance, and obtain harmonious facial aesthetic results so the successfulness of orthodontic treatment should be comprehensive because maloclusion can occur in dentoalveolar, skeletal and soft tissue tissues. Purpose: This study aims to evaluate the successfulness of orthodontic treatment using fixed orthodontic appliances in Orthodontic Clinic Dental Hospital, Faculty of Dental Medicine Universitas Airlangga by means of ABO system. Methods: Descriptive analytic by comparing the data before and after treatment, then the sample was divided into 3 based on the skeletal malocclusion group. Dental efficacy was measured using the ABO system and the Bolton anterior ratio. Skeletal success by looking at ANB, FHI, and proportion of anterior facial height (UAFH: LAFH). Meanwhile, the success of the soft tissue was by seeing the changes in the aesthetic lines of the upper and lower lips. The statistics used in this study were Kruskal-Whallis for the ABO DI and OGS difference test, Spearman to determine the relationship between ABO DI and OGS measurement components and McNemar and Wilcoxon to determine the difference before and after treatment on all measurements. Results: Treatment of class I skeletal malocclusion had the best mean ABO OGS score. Class II and III treatments had significant treatment advancements. In other measurements, there are significant differences after treatment at FHI in class I, (UAFH: LAFH) in class II and lower lip esthetic line in class II. Conclusion: Generally, the successfulness of orthodontic treatment in Orthodontic Clinic Dental Hospital, Faculty of Dental Medicine, Universitas Airlangga was adequate. In addition, the assessment of the successful orthodontic treatment needs to be adjusted to the standard values that can be accepted by a certain population, especially the Javanese population.

Changes of the redox-regulatory system in tumor and its surrounding tissues in patients with soft tissue sarcoma (STS) after neoadjuvant chemotherapy (NC).

e23522 Background: The disruptions in redox homeostasis in the nonmalignant tissues surrounding neoplasm can promote the tumor progression. The aim of this work was to assess the changes of the redox-regulatory system in the tumor and tumor-surrounding tissues in STS patients (pts) under the influence of a mofified metod of NC. Methods: The activity of glutathione S-transferase (GST), glutathione peroxidase (GPx), glutathione reductase (GR), content of reduced glutathione (GSH) and malondialdehyde (MDA) were determined by spectrophotometric methods. All markers were measured in the samples of tumor, peritumoral area and healthy tissues (taken along the line of resection) obtained during the surgery from 42 STS pts (T2a-bN0M0). The control group consisted of 21 primary pts who underwent resection only. Patients of the experimental group (21) received NC comprising systemic and local administration of antitumor drugs. Doxorubicin (40 mg/m2) was injected intravenously on the 1st and 7th days with autologous red blood cells as drug carriers; at the same time, cyclophosphamide (600 mg/m2) and methotrexate (40 mg/m2) were injected along the tumor periphery, on autologous plasma as a carrier. After 14 days, tumor removal surgery performed. All STS pts received standard postoperative chemoradiotherapy. Results: The level of GSH in tumor without NC treatment was higher than in the healthy and peritumoral tissue (by 2.3-2.5 times), and the activity of all glutathione-dependent enzymes was higher by 53.0-147.0 % (p = 0.0413-0.00124). The content of MDA in tumor was lower than in other tissues by 30.0-46.0 % (p = 0.00061). We did not find any differences between the healthy and peritumoral areas. In tumor samples of the experimental group, we also observed statistically significant increase in the level of GSH (by 2.8–3.0 times) and activation of GPO (by 37.3-95.8 %) and GR (by 2.0-3.2 times) vs. other tissues. However, after NC, the studied samples showed an increase in GSH by 3.1–3.8 times (p = 0.0143–0.00112), compared with the corresponding control samples. Also, the activity of GPO (by 54.5 %) and GsT (by 38.9 %) was significantly increased in peritumoral tissue vs similar area in the control group. After NC, the content of MDA was reduced in the healthy and tumor tissues vs control by 52.0 % (p = 0.0074) and 30.6 % (p = 0.04815), respectively. Clinical efficacy of NC was confirmed by reduced tumor volume in most patients by 30-40 %; the 5-year monitoring of STS pts showed that local recurrence and metastasis occurred in 14 of 21 pts in the control group, and in 6 of 21 in the main group (p = 0.0294). Conclusions: The NC treatment modifies the redox balance in the tumor-surrounding tissues and, as a result, decreases the oxidation damages in the healthy tissues. This effect, apparently, is an additional factor that improves the effectiveness of the proposed NC method.

Immunohistochemical localization of carbonic anhydrase IV in the human parotid gland

ABSTRACT Carbonic anhydrases (CAs) catalyze the hydration and dehydration of carbon dioxide. They are important for regulating ions, fluid and acid-base balance in many tissues. The location of CAs by cell type is important for understanding their roles in these functions. CAs II and VI have been demonstrated using immunohistochemistry (IHC) in the serous acinar cells of human salivary glands and ducts of rat salivary glands. CA IV has been localized by IHC to the ducts of rat salivary glands. CA IV also is present in human parotid glands as shown by real time-polymerase chain reaction (RT-PCR), but this method does not show the distribution of the CA isozymes by cell type. We investigated the cell-specific distribution of CA IV in the human parotid gland. Sections from five formalin fixed, paraffin embedded specimens of human parotid gland were subjected to IHC for CA IV using a commercial antibody. Moderate to strong reactions were found in the cell membranes and cytoplasm of the intercalated, striated and excretory ducts and capillaries, and reactions in the acini were limited to faint areas in some cells. These results indicate that CA IV participates in the regulation of bicarbonate/carbon dioxide fluxes in the ductal system of the human parotid gland.

The evolving cardiac lymphatic vasculature in development, repair and regeneration.

The lymphatic vasculature has an essential role in maintaining normal fluid balance in tissues and modulating the inflammatory response to injury or pathogens. Disruption of normal development or function of lymphatic vessels can have severe consequences. In the heart, reduced lymphatic function can lead to myocardial oedema and persistent inflammation. Macrophages, which are phagocytic cells of the innate immune system, contribute to cardiac development and to fibrotic repair and regeneration of cardiac tissue after myocardial infarction. In this Review, we discuss the cardiac lymphatic vasculature with a focus on developments over the past 5 years arising from the study of mammalian and zebrafish model organisms. In addition, we examine the interplay between the cardiac lymphatics and macrophages during fibrotic repair and regeneration after myocardial infarction. Finally, we discuss the therapeutic potential of targeting the cardiac lymphatic network to regulate immune cell content and alleviate inflammation in patients with ischaemic heart disease.

Characteristics of Trace Element Balance in Children with Pollution-Related Dental Diseases

Summary. Introduction: The necessity to assess, prevent and correct element-dependent diseases, increase the level of safety, predict and reduce the risk of exposures to industrial contaminants in the development of dental diseases in the child population is an important and promising area of modern hygiene. The objective of our study was to establish concentrations of trace elements in dental tissues of children exposed to various levels of industrial pollution. Materials and methods: We used atomic absorption spectrophotometry to measure 13 trace elements in extracted healthy and carious primary teeth in two groups of children aged 7–11. The first group consisted of 56 children living in the area with a significantly high level of anthropogenic burden (Caveraged total = 1.17 units) and a high prevalence of caries (86.9 %) while the second group consisted of 68 children living in the area with a relatively high level of anthropogenic burden (Caveraged total = 0.68 units) and a mean prevalence of caries (77.1 %) according to WHO criteria. Results: Our findings indicate that accumulation of toxic microelements in hard dental tissues of children in the first group was higher than in children of the second group: concentrations of lead, bismuth, cadmium and strontium were 2.6, 1.8, 2.5, and 1.2 times higher, respectively. We also observed higher accumulation of environmental toxicants in carious teeth compared to healthy ones. Thus, lead, cadmium and bismuth levels in carious teeth were 3.04, 1.2 and 3.13 times higher than in healthy teeth, respectively, while the level of strontium was, on the opposite, 2.5 times lower. Conclusions: The study revealed specific features of the interaction of essential trace elements in children in the form of antagonistic effects between cadmium and zinc, copper and manganese, and synergism in the content of copper, iron, nickel, zinc, and cobalt in healthy teeth, the mechanism of which is based on competition for a bond with a carrier substance and replacement of one element with another. In addition, we established that changes in the microelement balance in the hard tissues of healthy and carious teeth are not unidirectional in nature, thus reflecting complex interactions between the child’s organism and anthropogenic factors. The results proved that accumulation of toxic trace elements in hard dental tissues promotes caries development and causes a sharp decrease in the content of essential trace elements, especially chromium, manganese, iron, and copper. The study showed that hard dental tissues are a depot for lead, cadmium, bismuth, and strontium in children environmentally exposed to high levels of industrial chemicals. Features of developing an imbalance of biotic concentrations of essential elements and accumulating toxic microelements in children with pollution-related tooth pathology were also established.

Hypoxia-induced miR-92a regulates p53 signaling pathway and apoptosis by targeting calcium-sensing receptor in genetically improved farmed tilapia (Oreochromis niloticus).

miR-92a miRNAs are immune molecules that regulate apoptosis (programmed cell death) during the immune response. Apoptosis helps to maintain the dynamic balance in tissues of fish under hypoxia stress. The aim of this study was to explore the role and potential mechanisms of miR-92a in the liver of tilapia under hypoxia stress. We first confirmed that CaSR (encoding a calcium-sensing receptor) is a target gene of miR-92a in genetically improved farmed tilapia (GIFT) using luciferase reporter gene assays. In GIFT under hypoxia stress, miR-92a was up-regulated and CaSR was down-regulated in a time-dependent manner. Knocked-down CaSR expression led to inhibited expression of p53, TP53INP1, and caspase-3/8, reduced the proportion of apoptotic hepatocytes, and decreased the activity of calcium ions induced by hypoxia in hepatocytes. GIFT injected in the tail vein with an miR-92a agomir showed up-regulation of miR-92a and down-regulation of CaSR, p53, TP53INP1, and caspase-3/8 genes in the liver, resulting in lower serum aspartate aminotransferase and alanine aminotransferase activities under hypoxia stress. These findings suggest that stimulation of miR-92a interferes with hypoxia-induced apoptosis in hepatocytes of GIFT by targeting CaSR, thereby alleviating liver damage. These results provide new insights into the adaptation mechanisms of GIFT to hypoxia stress.

Acute effect of kinesiology taping on muscle strength, tissue temperature, balance, and mobility in female patients with osteoarthritis of the knee

Aim The data on the action mechanism of the kinesiology taping (KT) used to reduce pain and inflammation, provide mechanical support, and facilitate or inhibit muscles in the treatment of osteoarthritis (OA) of the knee are contradictory. The aim of this study was to investigate the acute effects of KT treatment on muscle strength, tissue temperature, balance, and mobility in female patients diagnosed with OA of the knee. Methods Thirty-four female patients (age: 60.26 ± 6.51 years) who were diagnosed with Kellgren–Lawrence Grade 2 OA according to the American Rheumatology Association (ACR-1986) criteria were included in the study. Patients who were performed KT with the superior Y, inferior Y, and U strip techniques were evaluated before and 30 min after taping. Muscle strength was assessed with a handheld dynamometer, tissue temperature with a thermal camera, balance with a monoaxial balance platform, and mobility the Timed Up and Go (TUG) test. Results After taping, there was a significant increase in the muscle strength during both extension and flexion (p<.001, p=.005). There was no statistically significant difference in the tissue temperature and balance scores before and after taping (p=.219, p>.05). There was a significant improvement in TUG scores, which evaluated mobility, after taping (p=.033). Conclusions Although KT treatment seems to be ineffective on tissue temperature and balance in the short term in patients with OA of the knee, we are of the opinion that patients should be included in the treatment program due to its positive effects on muscle strength and mobility.

ST Segment Yükselmeli Miyokard İnfarktüsü Hastalarında Koroner Arter Hastalığının Ciddiyeti ile Serum Adropin Seviyesi Arasındaki İlişkinin Değerlendirilmesi

Objective: Adropin is associated with energy balance in tissues and organs. The SYNTAX score (SS) is used to determine the severity of ST-segment elevation myocardial infarction (STEMI). The aim of this study was to determine the relationship between serum adropin levels and disease severity in STEMI patients. Materials and Methods: Eighty-nine patients who underwent coronary angiography (CAG) for STEMI were included in the study. The STEMI patients were divided into two subgroups: Group 1(SS < 22) and Group 2 (SS ≥ 23). 43 patients who underwent CAG and had normal coronary arteries were included in the study as a control group. Results: Groups 1 and 2 included more male participants than the control group (89.7% and 74.2% vs. 34.9%, respectively, p < 0.001). The smoking rate was higher in Groups 1 and 2 than in the control group (55.1% and 34.4% vs. 11.6%, respectively, p≤0.001). Serum adropin levels were lower in Group 1 than in the control group (147.3 ± 149.2 mg/L and 228.1 ± 253.3 ng/L, p = 0.03). Serum adropin levels were the lowest in Group 2 (87.8 ± 23.2 ng/L, 147.3 ± 149.2 ng/L, and 228.1 ± 253.3 ng/L, p = 0.004). Serum adropin levels were also negatively correlated with SS (r = -0.33, p = 0.002). Conclusion: Serum adropin levels decreased more in STEMI patients than in those without coronary artery disease (CAD). In addition, serum adropin levels decreased with increasing SS;this indicates the severity of CAD.

Прооксидантно-антиоксидантна рівновага в тканинах серця і мозку гусей за ембріонального та раннього постнатального онтогенезу

Переваги гусей породи Легарт зумовлюють достатньо широкий попит та їхнє швидке розповсюдження в господарствах країни. Втім, гуси цієї породи достатньо чутливі до незбалансованих кормів, особливо в перші тижні життя. Розуміння механізмів формування адаптивної відповіді на пошкоджуючий вплив сприятиме вірному вибору стратегії, спрямованої на регуляцію стану redox системи організму птиці та розкриття генетичного потенціалу продуктивності даної породи. Мета досліджень - з’ясування особливостей підтримки прооксидантно-антиоксидантної рівноваги в тканинах гусей породи Легарт під час ембріонального і раннього постнатального розвитку. Досліджено специфічність підтримки рівноваги між про- і антиоксидантами в тканинах серця та мозку гусей цієї породи. Доведено, що в мозку генетично запрограмована активізація антиоксидантної системи, спрямована на адаптацію організму гусей до гіпероксії початку атмосферного дихання, відбувається шляхом підвищення активності антиоксидантних ензимів і зниження вмісту ненасичених жирних кислот. У міокарді підтримка прооксидантно-антиоксидантної рівноваги здійснюється переважно за рахунок активізації ферментативної складової антиоксидантної системи. Втім, з урахуванням протилежно спрямованих змін коефіцієнта антиоксидантної активності і активності антиоксидантних ензимів з одного боку, а з іншого, стабільно високого вмісту ненасичених жирних кислот, саме в міокарді дуже висока ймовірність реалізації альтернативних механізмів антиоксидантного захисту.

STANDARDIZATION OF ROUTINE QUANTIFICATION OF POLYUNSATURATED ACIDS IN PEDIATRIC TOTAL PARENTERAL NUTRITION

Plasma and erythrocyte PUFAs are useful in clinical practice to optimize the parenteral nutrition therapy. The aim of this study was the definition of pediatric PUFAs reference ranges to follow-up patients with IICB. We validated a method to quantify, in plasma and erythrocyte membranes, glycerophospholipids PUFAs by gas chromatography coupled with the highly specific Mass spectrometer. The analyzed PUFAs are only those metabolized by liver: no interferences by oral nor parenteral nutrition are expected. Metabolically significative PUFAs were selected for analysis: ARA (ω6), EPA and DHA (ω3) were quantified because they are precursors of important mediators of inflammation, while Mead Acid determination is useful to calculate the ratio ARA (ω6)/ Mead (ω9), a known marker of essential fatty acid deficiency. We quantified these PUFAs in healthy pediatric subjects to build reference ranges. A pilot study was performed to compare PUFAs of two groups of pediatric patients with IICB (group A fed with Clinoleic® and group B with SMOF®) with reference ranges. Pediatric reference ranges of PUFAs were proposed. A good correlation between plasma and erythrocyte results was shown for the ratio ARA/(EPA+DHA), (R2=0.941). PUFAs assessment is highly recommended in plasma rather than in erythrocytes because plasma is less prone to peroxidation artifacts. Our fast method can offer an accurate, specific and easy-to-handle PUFAs analysis and a useful marker of w6/w3 balance in tissues for routine clinical practice. This work could be a step towards standardization of routine determination of PUFAs.

Effects of Prostaglandin Analogue Iloprost Treatment on Distant Organ Damage Following Ovarian Ischemia/Reperfusion Injury in Rats: An Experimental Study.

BackgroundIschemia/reperfusion (I/R) injury causes oxidative stress, which, in turn, may impair the oxidant/antioxidant balance in tissues and cause damage to the tissues. The local effects of I/R injury can be typically observed in the related organ while systemic effects can be observed predominantly in the heart, brain, lung, and kidney. In this study, we aimed to evaluate the effects of iloprost on heart tissues after an ovarian I/R injury in an experimental rat model.Materials and methodsA total of 32 female Sprague Dawley rats were used for the experiment. The rats were divided into four groups with eight rats each: Group I, control group; Group II, ischemia group; Group III, I/R group; Group IV, I/R + iloprost group. Surgical intervention was performed in each group and after the procedures, heart tissues were obtained and examined histopathologically.ResultsNo significant pathological finding was found in Group I and II while degeneration of muscle fibers and interstitial edema was observed in group III and dilation of the vessels was detected in Group IV. No fibrosis or inflammation was observed in any group.ConclusionIloprost provided protection against I/R injury and thus may be an alternative treatment for I/R injury.

Age-Related Effects of Orthovanadate Nanoparticles Involve Activation of GSH-Dependent Antioxidant System in Liver Mitochondria.

Vanadium is an important ultra-trace element nowadays attracting attention with particular emphasis on medical application. But the therapeutic application of vanadium-based drugs is still questionable and restricted due to some toxic side effects. It was found that unique redox properties of vanadium in nanoform provided antioxidant activity and prevented oxidative disturbance in cells in vitro. Though, on the organism level, ambiguous effects of vanadium-based nanoparticles were observed. In this study, the age-related features of prooxidant/antioxidant balance in blood serum and liver mitochondrial and postmitochondrial fractions of 3 and 18-month-old Wistar male rats treated with orthovanadate nanoparticles (GdVO4/Eu3+, 8 × 25nm) within 2months have been investigated. Prooxidant potential-related indexes were the content of lipid hydroperoxides as well as aconitase activity. Activity of glutathione peroxidase, glutathione-S-transferase, glutaredoxin, glutathione reductase, glucose-6-phosphate dehydrogenase, and NADPH-dependent isocitrate dehydrogenase designated the tissue antioxidant potential. Based on the obtained values, the integral index of the prooxidant/antioxidant balance-the reliability coefficient (Kr) has been calculated. The data show that due to activation some chain links of GSH-dependent antioxidant system, GdVO4/Eu3+ nanoparticles increase the reliability of the prooxidant-antioxidant balance in tissues and especially in the liver mitochondria of old animals (Kr in mitochondria of young rats was 2.94, and in mitochondria of old ones-9.83 conventional units). Detected in vitro glutathione peroxidase-like activity of the GdVO4/Eu3+ nanoparticles is supposed to be among factors increasing the reliability of the system. So, for the first time, the beneficial effect of the long-term orthovanadate nanoparticle consumption in old males has been discovered.

Immunotherapy for Upper Airway Disease Generates IL-10-Producing KLRG1 &lt;sup&gt;+&lt;/sup&gt; ILC2s that are Associated with Clinical Benefit

SummaryIn this study, we addressed the role of IL-10-producing ILCs in controlling grass pollen allergy. IL-10+ ILCs are comprised in KLRG1+ILC2-lineage cells and absent on CRTH2+ ILC2 which lack KLRG1. We observed that ILC2s in healthy subjects and grass pollen allergic patients show differential gene expression profiles. A cross-sectional study demonstrates that IL-10-producing KLRG1+ ILC2s are strongly reduced in patients with grass pollen and house dust mite allergy when compared to healthy subjects and those who have received grass pollen subcutaneous immunotherapy treatment. In a prospective double-blind, placebo-controlled trial of grass pollen sublingual immunotherapy (SLIT), we observed that the capacity of ILC2 to produce IL-10 was restored in patients who received SLIT, but not patients receiving a placebo. Finally, we demonstrated that these IL-10-producing ILCs are functional and play a crucial role in restoring barrier function and tissue balance in allergic diseases.