Changes In Cell Permeability Research Articles

The Role of Cardio-Renal Inflammation in Deciding the Fate of the Arteriovenous Fistula in Haemodialysis Therapy.

Vascular access is an indispensable component of haemodialysis therapy for end-stage kidney disease patients. The arteriovenous fistula (AVF) is most common, but importantly, two-year failure rates are greater than fifty percent. AVF failure can occur due to a lack of suitable vascular remodelling, and inappropriate inflammation preventing maturation, or alternatively neointimal hyperplasia and vascular stenosis preventing long-term use. A comprehensive mechanistic understanding of these processes is still lacking, but recent studies highlight an essential role for inflammation from uraemia and the AVF itself. Inflammation affects each cell in the cascade of AVF failure, the endothelium, the infiltrating immune cells, and the vascular smooth muscle cells. This review examines the role of inflammation in each cell step by step and the influence on AVF failure. Inflammation resulting in AVF failure occurs initially via changes in endothelial cell activation, permeability, and vasoprotective chemokine secretion. Resultingly, immune cells can extravasate into the subendothelial space to release inflammatory cytokines and cause other deleterious changes to the microenvironment. Finally, all these changes modify vascular smooth muscle cell function, resulting in excessive and unchecked hyperplasia and proliferation, eventually leading to stenosis and the failure of the AVF. Finally, the emerging therapeutic options based off these findings are discussed, including mesenchymal stem cells, small-molecule inhibitors, and far-infrared therapies. Recent years have clearly demonstrated a vital role for inflammation in deciding the fate of the AVF, and future works must be centred on this to develop therapies for a hitherto unacceptably underserved patient population.

GMPR2-MDA5 interaction bridged the cell junction and anti-GCRV immune

As a consequence of increased vascular endothelial permeability and vascular injury caused by GCRV (grass carp reovirus) infection, grass carp (Ctenopharyngodon idellus) could suffer from hemorrhagic disease and even death. MDA5 is a key RNA sensor that stimulates type I interferon as well as regulates vascular permeability. The guano monophosphate ductase 2 (GMPR2) plays a key role in regulating the permeability of vascular endothelial cells. This study examined the interaction between grass carp MDA5 and GMPR2 and their antiviral effects. GMPR2 interacted directly with MDA5 and promoted its protein level, whilst MDA5 conversely inhibited the GMPR2 protein expression. The expression of GCRV VP7 protein was inhibited by the increased levels of MDA5 and GMPR2. In addition, the cell junction molecule Claudin7b which is closely related to the change in vascular permeability, was inhibited by both MDA5 and GMPR2. Finally, in GMPR2 knockout zebrafish, the mRNA level of MDA5 was significantly down-regulated, which validated the positive functional relationship of GMPR2 on MDA5. Consistent changes of the Claudin 7b and VP7 protein levels were observed in the GMPR2 and MDA5 overexpression groups. In summary, MDA5-GMPR2 interaction affecting GCRV replication might through cell junction molecule Claudin7b. The results from this study provided systematic support for revealing the genetic basis of cell permeability change as well as the hemorrhagic diseases in grass carp.

Cytoprotective 3K3A-activated protein C and plasma: A comparison of therapeutics for the endotheliopathy of trauma.

Both healthy plasma and cytoprotective aPC (3K3A-aPC) have been shown to mitigate the endotheliopathy of trauma (EoT), but optimal therapeutics remain unknown. Our aim was therefore to determine optimal therapies to mitigate EoT by investigating the effectiveness of 3K3A-aPC with and without plasma-based resuscitation strategies. Electric cell-substrate impedance sensing (ECIS) was used to measure real-time permeability changes in endothelial cells. Cells were treated with a 2 μg/mL solution of aPC 30 minutes prior to stimulation with plasma taken from severely injured trauma patients (ISS > 15 and BD < -6) (TP). Healthy plasma, or plasma frozen within 24 hours (FP24), was added concomitantly with TP. Cells treated with thrombin and untreated cells were included in this study as control groups. A dose-dependent difference was found between the 5% and 10% plasma-treated groups when HUVECs were simultaneously stimulated with TP (μd 7.346 95%CI 4.574 to 10.12). There was no difference when compared to TP alone in the 5% (μd 5.713 95%CI -1.751 to 13.18) or 10% group (μd -1.633 95%CI -9.097 to 5.832). When 3K3A-aPC was added to plasma and TP, the 5% group showed improvement in permeability compared to TP alone (μd 10.11 95%CI 2.642 to 17.57), but there was no difference in the 10% group (μd -1.394 95%CI -8.859 to 6.070). The combination of 3K3A-aPC, plasma, and TP at both the 5% plasma (μd -28.52 95%CI-34.72 to -22.32) and 10% plasma concentrations (μd -40.02 95%CI -46.22 to -33.82) had higher inter-cellular permeability than the 3K3A-aPC pre-incubation group. Our data shows that FP24, in a post-trauma environment, pre-treatment with 3K3A-aPC can potentially mitigate the EoT to a greater degree than FP24 with or without 3K3A-aPC. Although further exploration is needed, this represents a potentially ideal and perhaps superior therapeutic treatment for the dysregulated thromboinflammation of injured patients. Prognostic/Epidemiological, Therapeutic/Care Management, Level III.

MnO2-based capacitive system enhances ozone inactivation of bacteria by disrupting cell membrane

The application of ozone (O3) disinfection has been hindered by its low solubility in water and the formation of disinfection by-products (DBPs). In this study, capacitive disinfection is applied as a pre-treatment for O3 oxidation, in which manganese dioxide with a rambutan-like hollow spherical structure is used as the electrode to increase the charge density on the electrode surface. When a voltage is applied, the negative-charged microbes are attracted to the electrodes and killed by electrical interactions. The contact between microbes and capacitive electrodes leads to changes in cell permeability and burst of reactive oxygen species, thereby promoting the diffusion of O3 into the cells. After O3 penetrates the cell membrane, it can directly attack the cytoplasmic constituents, accelerating fatal and irreversible damage to pathogens. As a result, the performance of the capacitance-O3 process is proved better than the direct sum of the two individual process efficiencies. The design of capacitance-O3 system is beneficial to reduce the ozone dosage and DBPs with a broader inactivation spectrum, which is conducive to the application of ozone in primary water disinfection.

Induction of Programmed Cell Death in Acanthamoeba culbertsoni by the Repurposed Compound Nitroxoline.

Acanthamoeba is a ubiquitous genus of amoebae that can act as opportunistic parasites in both humans and animals, causing a variety of ocular, nervous and dermal pathologies. Despite advances in Acanthamoeba therapy, the management of patients with Acanthamoeba infections remains a challenge for health services. Therefore, there is a need to search for new active substances against Acanthamoebae. In the present study, we evaluated the amoebicidal activity of nitroxoline against the trophozoite and cyst stages of six different strains of Acanthamoeba. The strain A. griffini showed the lowest IC50 value in the trophozoite stage (0.69 ± 0.01 µM), while the strain A. castellanii L-10 showed the lowest IC50 value in the cyst stage (0.11 ± 0.03 µM). In addition, nitroxoline induced in treated trophozoites of A. culbertsoni features compatibles with apoptosis and autophagy pathways, including chromatin condensation, mitochondrial malfunction, oxidative stress, changes in cell permeability and the formation of autophagic vacuoles. Furthermore, proteomic analysis of the effect of nitroxoline on trophozoites revealed that this antibiotic induced the overexpression and the downregulation of proteins involved in the apoptotic process and in metabolic and biosynthesis pathways.

Intravenous Injection of Na Ions Aggravates Ang II-Induced Hypertension-Related Vascular Endothelial Injury by Increasing Transmembrane Osmotic Pressure.

Extracellular protein nanoparticles (PNs) and ions perform synergistical functions in the control of transmembrane osmotic pressure (OP) under isotonic conditions. Intravenous injection may disrupt the ion balance and alter PN levels in blood plasma, changing transmembrane OP and damaging vascular endothelial cells. Na ions were injected into AngII-induced HUVECs to simulate cell injury in vitro, and tail vein infusion of Na ions into hypertensive rats was performed to assess vascular damage. Optical measurements using an intermediate filament (IF) tension probe were conducted to detect indicators related to transmembrane OP. Immunofluorescence, Western blotting and small interfering RNA (siRNA) transfection were employed to investigate inflammasomes and the relationship between Abl2 and inflammation. Electrolyte injections with sodium ions (but not glucose and hydroxyethyl starch) induced the production of ASC and NLRP3 inflammasomes in Ang II-induced HUVECs; this in turn resulted in the disorder of calcium signals, and changes in transmembrane OP and cell permeability. Moreover, injection of Na ions into Ang II-induced HUVECs activated the mechanosensitive protein Abl2, involved in inflammation-induced transmembrane OP changes. A drug combination was identified that could induce OP recovery and block hyperpermeability induced by cytoplasmic inflammatory corpuscles in vivo and in vitro. Changes in extracellular PNs and ions following chemical stimuli (Ang II) participate in the regulation of transmembrane OP. Furthermore, injection of Na ions causes vascular endothelial injury in Ang II-induced cells in vitro and hypertension rats in vivo, suggesting it is not safe for hypertensive patients, and we propose a new drug combination as a solution.

Semaphorin 7a regulates inflammatory mediators and permeability in retinal endothelial cells

Research supports a key role for inflammation in damaging the retinal vasculature. Current work is designed to investigate regulation of key inflammatory pathways. In this study, we hypothesized that semaphorin 7a (Sema7a) was involved in the increased inflammatory mediators and permeability changes in retinal endothelial cells (REC) grown in high glucose. For these studies, we used diabetic mouse samples and REC to investigate our hypothesis. Primary retinal endothelial cells were grown in normal (5 mM) or high glucose (25 mM glucose) for measurements. In a subset of cells grown in high glucose, cells were transfected with Sema7a siRNA or scrambled siRNA. We measured levels of key inflammatory mediators and zonula occludens-1 (ZO-1) and occludin levels by Western blot. Data suggest that high glucose increased inflammatory mediators and reduced the tight junction proteins, which follows what is often observed in cells grown in high glucose. Sema7a siRNA significantly decreased inflammatory proteins and increased levels of ZO-1 and occludin. These data suggest that Sema7a mediates the actions of high glucose in REC. Use of Sema7a siRNA may offer a new avenue for treatment.

Realistic numerical simulations of diffusion tensor cardiovascular magnetic resonance: The effects of perfusion and membrane permeability.

To study the sensitivity of diffusion tensor cardiovascular magnetic resonance (DT-CMR) to microvascular perfusion and changes in cell permeability. Monte Carlo (MC) random walk simulations in the myocardium have been performed to simulate self-diffusion of water molecules in histology-based media with varying extracellular volume fraction (ECV) and permeable membranes. The effect of microvascular perfusion on simulations of the DT-CMR signal has been incorporated by adding the contribution of particles traveling through an anisotropic capillary network to the diffusion signal. The simulations have been performed considering three pulse sequences with clinical gradient strengths: monopolar stimulated echo acquisition mode (STEAM), monopolar pulsed-gradient spin echo (PGSE), and second-order motion-compensated spin echo (MCSE). Reducing ECV intensifies the diffusion restriction and incorporating membrane permeability reduces the anisotropy of the diffusion tensor. Widening the intercapillary velocity distribution results in increased measured diffusion along the cardiomyocytes long axis when the capillary networks are anisotropic. Perfusion amplifies the mean diffusivity for STEAM while the opposite is observed for short diffusion encoding time sequences (PGSE and MCSE). The effect of perfusion on the measured diffusion tensor is reduced using an increased reference b-value. Our results pave the way for characterization of the response of DT-CMR to microstructural changes underlying cardiac pathology and highlight the higher sensitivity of STEAM to permeability and microvascular circulation due to its longer diffusion encoding time.

Effect of Low Temperature on Photosynthetic Physiological Activity of Different Photoperiod Types of Strawberry Seedlings and Stress Diagnosis

During the early growth stage of plants, low temperatures can alter cell permeability, reduce photosynthetic capacity, and have adverse effects on crop growth, development, and yield. Different strawberry cultivars have varying cold tolerance. In this study, we investigated the changes in cell permeability and photosynthetic activity of short-day and long-day types of strawberry cultivars under varying degrees of low-temperature stress, and evaluated the extent of cellular damage using photosynthetic and chlorophyll fluorescence parameters. The experiment utilized short-day strawberry cultivars ‘Toyonoka’ and ‘Red Face’, and long-day strawberry cultivars ‘Selva’ and ‘Sweet Charlie’ seedlings. Low-temperature treatments were set at −20, −15, −10, −5, 0, 5, and 10 °C for 12 h. The research demonstrated that short-day strawberries had greater tolerance to low temperatures, and all four strawberry cultivars began to experience low-temperature stress when the temperature was below 5 °C. A temperature range of 0 to −10 °C played a crucial role in causing severe cold damage to the strawberries. The low-temperature stress levels were constructed based on electrolyte leakage, with photosynthetic physiological characteristics serving as references. The study proves that the photosynthetic and chlorophyll fluorescence parameters can serve as effective probes for diagnosing low-temperature stress in strawberry seedlings, and their combination provides higher accuracy in identifying stress levels than any single type of parameter.

Improved efficacy of eugenol and trans-anethole in combination with octenidine dihydrochloride against Candida albicans and Candida parapsilosis.

Candidiasis is a fungal infection caused by yeasts from the Ogenus Candida. Considering increasing antifungal resistance rates the activity was analyzed of natural compounds to eradicate Candida spp. The aim of the study was to check the antifungal activity of selected essential oil compounds (EOCs; thymol, menthol, eugenol [E], carvacrol, trans-anethole [TA]) alone, and in combination with octenidine dihydrochloride (OCT) against C.albicans and C.parapsilosis reference, and clinical strains. Investigated clinical isolates were obtained from skin wounds of patients treated for superficial wounds candidiasis. The following parameters were studied: antifungal susceptibility testing using the VITEK system, antifungal activity of EOCs alone and in combination with OCT using microdilution and checkerboard assays, antifungal efficacy of selected chemicals using time-kill curve assay, and changes in cell permeability in the presence of selected chemicals using crystal violet assay. Clinical isolates of C.albicans and C.parapsilosis were resistant to fluconazole and voriconazole. The highest inhibition activity against Candida isolates was observed for E.The OCT - TA and OCT - E combinations showed synergistic and additive activities against all strains, respectively. These combinations also appeared to affect the rate of yeast cell killing and increasing the permeability of Candida cells. The study indicates that E and TA potentially used in formulation with OCT might eradicate pathogenic yeasts; however, microbiological and clinical studies are still required.

Cytotoxicity of some preservatives for cultureChlorella VulgarisGKO strain

One of the ways to preserve the culture of microalgae for a long time is conservation. All modern methods of preservation and long-term storage of microorganism cultures are based on the transfer of cells to a state of anabiosis, in which metabolic processes are either completely or partially stopped. One of the mechanisms that are realized during the transition to the anabiotic state is a change in cell permeability. This can be achieved by modifying the culture medium. Thus, the search for preservatives that contribute to the long-term preservation of biomass is very relevant. The primary condition for the use of any preservative or a new component of the culture medium should be the absence of a toxic effect on the cell culture. Goal of the work: to determine the cytotoxic effect of some preservatives for the cell culture ofChlorella vulgarisGKO strain. The cytotoxicity of ascorbic acid, lactic acid, citric acid, acetic acid, sodium chloride and urotropin was analyzed for the cell culture ofChlorella vulgarisstrain GKO. The following parameters were determined: pH ofChlorella vulgarissuspension with preservative; Total number of cells, MM/ml; The ratio of dead cells to the total number of cells,%; Specific growth/death rate; The optical density of the suspension; The difference in the average optical density,%; Cell size (diameter), µm pH of suspension ofChlorella vulgariswith preservative; Total number of cells, MM/ml; The ratio of dead cells to the total number of cells,%; Specific growth/death rate; The optical density of the suspension; The difference in the average optical density,%; Cell size (diameter), µm pH of suspension ofChlorella vulgariswith preservative; Total number of cells, MM/ml; The ratio of dead cells to the total number of cells,%; Specific growth/death rate; The optical density of the suspension; The difference in the average optical density,%; Cell size (diameter), µm

Do mast cells contribute to the continued survival of vertebrates?

This study is an attempt to shed light on why the connective tissue mast cell (MC) is preserved in all species with a blood circulatory system, i.e., the vertebrates since >500 million years, which suggests that the MC performs as yet not understood indispensible life‐promoting actions. The literature survey focuses on data in published papers on MC functions in immunological and nonimmunological reactions, host protection, pregnancy, inflammation, and wound healing. All data are thus accessible to the reader. The MC is a secretory cell with a unique mediator profile. A distinctive role for MCs is defined not only by their extensive mediator composition but also by their prominent ability to affect the vasculature to expedite selective cell recruitment and permeability changes and to set the stage for an appropriate acquired response. MCs, harboring a wide range of surface membrane receptors, are activated by the major female sex hormones as well as by diverse potentially adverse stimuli. MC activation/degranulation creates a presumably unique triad tissue response in physiological and pathological situations alike: extracellular matrix degradation and tissue remodeling, de novo cell proliferation, and de novo angiogenesis. As shown in the literature, MC‐activation is crucial for successful female reproduction in the mouse, implying one of possibly several yet unidentified physiological roles of MCs. Moreover, the activated MC aids newborns to survive to reproductive age owing to its key beneficial actions in inflammation and wound healing. Thus, a not previously described life‐perpetuating loop spanning generations are apparently formed, which, hypothetically, could contribute to the continued survival of the vertebrates.

Determining Intestinal Permeability using Lucifer Yellow in an Apical-Out Enteroid Model.

Enteroids are an emerging research tool in the study of inflammatory bowel diseases such as necrotizing enterocolitis (NEC). They are traditionally grown in the basolateral-out (BO) conformation, where the apical surface of the epithelial cell faces the inner lumen. In this model, access to the luminal surface of enteroids for treatment and experimentation is challenging, which limits the ability to study host-pathogen interactions. To circumvent this, a neonatal apical-out (AO) model for necrotizing enterocolitis was created. Since intestinal epithelial cell permeability changes are pathognomonic for NEC, this protocol outlines using lucifer yellow (LY) as a marker of paracellular permeability. LY traverses the intestinal epithelial barrier via all three major paracellular pathways: pore, leak, and unrestricted. Using LY in an AO model allows for a broader study of permeability in NEC. Following IRB approval and parental consent, surgical samples of intestinal tissue were collected from human preterm neonates. Intestinal stem cells were harvested via crypt isolation and used to grow enteroids. Enteroids were grown to maturity and then transformed AO or left in BO conformation. These were either not treated (control) or were treated with lipopolysaccharide (LPS) and subjected to hypoxic conditions for the induction of in vitro NEC. LY was used to assess for permeability. Immunofluorescent staining of the apical protein zonula occludens-1 and basolateral protein β-catenin confirmed AO conformation. Both AO and BO enteroids treated with LPS and hypoxia demonstrated significantly increased paracellular permeability compared to controls. Both AO and BO enteroids showed increased uptake of LY into the lumen of the treated enteroids compared to controls. The utilization of LY in an AO enteroid model allows for the investigation of all three major pathways of paracellular permeability. It additionally allows for the investigation of host-pathogen interactions and how this may affect permeability compared to the BO enteroid model.

Effect analysis on pressure sensitivity performance of diesel particulate filter for heavy-duty truck diesel engine by the nonlinear soot regeneration combustion pressure model

In this paper, the nonlinear soot regeneration combustion pressure model (NSRCMP model) is established and employed for Diesel Particulate Filter (DPF) simulation study. The NSRCMP model is reliable and accurate under simulation and experimental conditions of the process of clean filter, soot loading and soot regeneration combustion. The DPF soot loading response time and pressure drop are investigated under the parameter changes of cell density, wall thickness and permeability. It is a new attempt to measure the DPF pressure sensitivity by the trend of different design parameters. Cell density and permeability affect the soot loading time dramatically and the maximum and minimum soot loading time ratio is 3. Cell density and permeability make a significant pressure drop change through the pressure sensitivity comparisons. The corresponding ratio of maximum pressure sensitivity parameter is 2.4 when the cell density is 100 cpsi. The comparison results of pressure sensitivity under different permeabilities show a fixed ratio of 1.4. The results show that cell density is the key parameter to affect the overall system pressure. The influence of wall thickness on pressure sensitivity is not sensitive to the change of configurations of DPFs.

MicroRNA-466 and microRNA-200 increase endothelial permeability in hyperglycemia by targeting Claudin-5

Endothelial cell (EC) permeability is essential to vascular homeostasis in diabetes. MicroRNAs are critical gene regulators whose roles in the EC permeability have yet to be characterized. This study aims to examine the change in cell permeability induced by miR-200 and miR-466 in ECs. Human aortic ECs and dermal microvascular ECs from healthy subjects and type 2 diabetic patients were used. Our in vitro experiments unveiled higher expressions of miR-200 family members and miR-466 in diabetic ECs and in healthy ECs when exposed to high glucose. Overexpression of both miR-200 and miR-466 significantly increased EC permeability through transcriptional suppression of Claudin-5, the cell tight junction protein, by directly binding to its 3′ untranslated region. In a mouse model of chronic hyperglycemia mimicking type 2 diabetes in humans (db/db mice), the delayed closure rate of a full-thickness excisional wound was partly rescued by topical application of the miR-200 inhibitor. The topical application of both miR-200 and miR-466 inhibitors exhibited improved efficacy in accelerating wound closure compared with the topical application of miR-200 inhibitor alone. Our study demonstrated the potentially effective approach of miR-200/miR-466 cocktail inhibition to restore vascular integrity and tissue repair in hyperglycemia.

Antibacterial activity and mechanism of essential oils in combination with medium-chain fatty acids against predominant bovine mastitis pathogens.

Bovine mastitis has become a significant economic importance for the dairy industry. Concerns regarding poor milk quality and emergence of bacterial resistance have necessitated to develop an alternative therapeutic approach to antibiotics for the treatment of mastitis. Saturated medium-chain fatty acids (MCFAs) and essential oils (EOs) are known natural antimicrobials, but their combined effect has not been investigated extensively. The objective of the present investigation was to examine the bactericidal effect of various combined treatments of eight EOs and three saturated MCFAs to inactivate predominant mastitis pathogens, including Staphylococcus aureus ATCC 29213; Escherichia coli ATCC 25922; Klebsiella pneumoniae ATCC 27736 and Streptococcus agalactiae ATCC 27956. The minimum inhibitory concentration (MIC) values confirmed that all the tested pathogens were variably susceptible to both EOs and saturated MCFAs. Among essential oils, carvacrol (CAR), trans-cinnamaldehyde (TC) and thymol (TM) showed the highest inhibitory activity at concentration 0·38-1·32 mg/mL. Carvacrol exhibited effective additive antibacterial activity in combined treatment with octanoic acid (OA) in terms of its fractional inhibitory index (0·63-0·88) and time-kill effect in reducing about 6 log CFU/mL bacterial cells in less than 5 min. The effort was also made to elucidate the mechanism of antibacterial action of CAR and OA against selected mastitis pathogens by observing changes in cell microstructure, permeability and integrity of cell membrane and their membrane potential. After adding CAR and OA at MIC level, there were obvious changes in cell morphology, leakage of small electrolytes and macromolecules at the initial few hours of treatment i.e. within 1-2 h were observed. Our results indicated that CAR and OA could be evaluated as alternatives or adjuncts to antibiotics as intramammary infusion or topical application to treat bovine mastitis, significantly improving the microbiological safety of milk.

Primary stress response and biochemical profile of Labeo rohita (Hamilton, 1822) experimentally parasitized with Argulus bengalensis (Ramakrishna, 1951).

Argulosis is a major problem that causes huge economic loss in aquaculture. In a microcosm, an infested condition was developed upon Labeo rohita with 100 ± 10 adult morphs of Argulus bengalensis per fish. Primary stress response and biochemical profiles of the host were evaluated to underscore the pathogenicity of the parasites. Significant alterations in biochemical parameters were monitored at four different post-infestation time points: days 1, 3, 6 and 9. The overall increasing trends of both plasma cortisol and plasma epinephrine indicate parasite-induced primary stress response among experimental fish. The study revealed a hyperglycaemic trend throughout the infestation period, which has been correlated with hypoxia-associated glycogenolysis. Decreasing level of plasma cholesterol has also been correlated with the development of anaemia and subsequent hypoxia among the infested fish. Plasma protein of the experimental fish initially increases as an outcome of the immediate innate immune response against Argulus infestation, whereas the decrease in plasma protein at the later period of infestation results from less-dietary protein intake due to loss of appetite, reduced digestibility and metabolic proteolysis. Plasma Na+ concentration showed an overall decreasing trend throughout the infestation period, which may be due to excess production of catecholamine under stress. Nonetheless, plasma K+ concentration showed an increasing trend up to day 6 of infestation, and thereafter the value declined to the control level. Plasma ionic imbalances reflect changes in cell permeability under tissue hypoxia and the wounds produced on the skin for intensive feeding activity of the parasites. Both serum glutamate oxaloacetate transaminase and serum glutamate pyruvate transaminase activities were significantly elevated throughout the infestation period, which indicates excess synthesis or release of those enzymes from the damaged cells of the tissues. Activities of some renal, hepatic and branchial antioxidant enzymes, viz., superoxide dismutase, catalase and glutathione-S-transferase, increased in the infested fish. The fact is explained as an effort of the fish for gaining adaptive adjustment to neutralize the oxidative stress generated under the parasitic stress. The overall experimental result points towards the generation of potential stress upon host fish by this branchiuran parasite. The biochemical alterations of the fish under argulosis are centred around the two stress-sensitive hormones, cortisol and epinephrine. The outcome of the study will be the important physiological determinants in adopting a suitable control measure as well as assessing the nutritional value of the fish under diseased condition.

SPAK Deficiency Attenuates Chemotherapy-Induced Intestinal Mucositis.

IntroductionSte20-related protein proline/alanine-rich kinase (SPAK) affects cell proliferation, differentiation, and transformation, and sodium and chloride transport in the gut. However, its role in gut injury pathogenesis is unclear.ObjectiveWe determined the role of SPAK in chemotherapy-induced intestinal mucositis using in vivo and in vitro models.MethodsUsing SPAK-knockout (KO) mice, we evaluated the severity of intestinal mucositis induced by 5-fluorouracil (5-FU) by assessing body weight loss, histological changes in the intestinal mucosa, length of villi in the small intestine, pro-inflammatory cytokine levels, proliferative indices, and apoptotic indices. We also evaluated changes in gut permeability and tight junction-associated protein expression. Changes in cell permeability, proliferation, and apoptosis were assessed in SPAK siRNA-transfected 5FU-treated IEC-6 cells.Results5-FU-treated SPAK-KO mice exhibited milder intestinal mucositis, reduced pro-inflammatory cytokine expression, increased villus length, good maintenance of proliferative indices of villus cells, decreased apoptotic index of enterocytes, reduced gut permeability, and restoration of tight junction protein expression (vs. 5-FU-treated wild-type mice). Under in vitro conditions, siRNA-mediated SPAK-knockdown in IEC-6 cells decreased cell permeability and maintained homeostasis following 5-FU treatment.ConclusionSPAK deficiency attenuated chemotherapy-induced intestinal mucositis by modulating gut permeability and tight junction-associated protein expression and maintaining gut homeostasis in murine small intestinal tissues following gut injury. The expression of SPAK may influence the pathogenesis of chemotherapy-induced intestinal mucositis.

Ionotropic Glutamate Receptors and Their Implications in Cancer and Cancer Therapeutics

Glutamine, an excitatory neurotransmitter, is necessary for physiological as well as pathological processes. Other than neuronal disorders and/or cancers, glutamate receptors have also been associated with an array of other malignancies. The metabotropic glutamate receptor (mGluR 1–8 [like Groups I, II, and III]) and ionotropic glutamate receptor (iGluR) have been targeted to treat cancers like carcinoma of the lung, breast, prostate, and oral cancer. iGluRs present on N-methyl-D-aspartate (NMDA) and non-NMDA receptors are multisubunit complexes. Since these subunits of NMDA receptors influence the mTOR signaling pathway significantly, their antagonists such as memantine, ifenprodil, or diclozipine are often used in cancer chemotherapy. Non-NMDA receptors such as α-amino 3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA) and kainate undergo glutamine to arginine site-specific RNA editing inflicting changes in cancer cell permeability. Thus, the employment of antagonists specific to these receptors would provide an effective anticancer therapeutic approach. Since AMPA receptors and kainate receptors have a crucial role in neural development and other cellular processes, their contribution in tumorigenesis has been mainly recognized in brain tumors although their role in further cancers cannot be ruled out. Delta or orphan receptors are primarily classified based on sequence homology. The effect and activity of antagonists for metabotropic and iGluRs have been pointed out due to their remedial contribution in various tumors. This review also highlights the relation of a range of subunits to cancer and anticancer agents as curatives for future applications and investigations.

Apoptosis-like cell death upon kinetoplastid induction by compounds isolated from the brown algae Dictyota spiralis

BackgroundThe in vitro activity of the brown seaweed Dictyota spiralis against both Leishmania amazonensis and Trypanosoma cruzi was evaluated in a previous study. Processing by bio-guided fractionation resulted in the isolation of three active compounds, classified as diterpenes. In the present study, we performed several assays to detect clinical features associated to cell death in L. amazonensis and T. cruzi with the aim to elucidate the mechanism of action of these compounds on parasitic cells.MethodsThe aims of the experiments were to detect and evaluate specific events involved in apoptosis-like cell death in the kinetoplastid, including DNA condensation, accumulation of reactive oxygen species and changes in ATP concentration, cell permeability and mitochondrial membrane potential, respectively, in treated cells.ResultsThe results demonstrated that the three isolated diterpenes could inhibit the tested parasites by inducing an apoptosis-like cell death.ConclusionsThese results encourage further investigation on the isolated compounds as potential drug candidates against both L. amazonensis and T. cruzi.Graphical