Effects of activin A and its downstream ERK1/2 in oxygen and glucose deprivation after isoflurane-induced postconditioning

BackgroundIsoflurane postconditioning (ISPOC) plays a neuroprotection role in the brain. Previous studies confirmed that isoflurane postconditioning can provide better protection than preconditioning in acute hypoxic–ischemic brain damage, such as acute craniocerebral trauma and ischemic stroke. Numerous studies have reported that activin A can protect rat’s brain from cell injury. However, whether activin A and its downstream ERK1/2 were involved in isoflurane postconditioning-induced neuroprotection is unknown.MethodsA total of 80 healthy Sprague–Dawley rats weighing 50–70 g were randomly divided into 10 groups of 8: normal control, oxygen and glucose deprivation (OGD), 1.5% ISPOC, 3.0% ISPOC, 4.5% ISPOC, blocker of activin A (SB431542), blocker of ERK1/2 (U0126), 3.0% ISPOC + SB431542, 3.0% ISPOC + U0126, and vehicle (dimethyl sulfoxide(DMSO)) group. Blockers (SB431542 and U0126) were used in each concentration of isoflurane before OGD. Hematoxylin–eosin staining, 2,3,5-triphenyl tetrazolium chloride staining, and propidium iodide (PI) staining were conducted to assess the reliability in the brain slices. Immunofluorescence, Western blot, and quantitative real-time PCR(Q-PCR) were performed to validate the protein expression levels of activin A, Smad2/3, P-Smad2/3, ERK1/2, and phosphorylation ERK1/2 (P-ERK1/2).ResultsThe number of damaged neurons and mean fluorescence intensity(MFI) of PI staining increased, but formazan generation, expression levels of activin A and P-ERK1/2 protein, and mRNA synthesis level of activin A decreased in the OGD group compared with the normal control group (p < 0.05). The number of damaged neurons and MFI of PI staining decreased, but formazan production, expression levels of activin A, P-Smad2/3, and P-ERK1/2, and mRNA synthesis level of activin A increased significantly in the 1.5% ISPOC and 3.0% ISPOC groups (p < 0.05) compared with the OGD group. The result in the 4.5% ISPOC group, was completely opposite to the 1.5% ISPOC and 3.0% ISPOC groups. The number of damage neuron and MFI of PI staining increased, but formazan production, expression levels of activin A, P-Smad2/3, and P-ERK1/2, and mRNA synthesis level of activin A decreased in the 4.5% ISPOC group. However, the expression levels of activin A, P-Smad2/3, and P-ERK1/2, and mRNA synthesis level of activin A in the 4.5% ISPOC group were higher than the OGD group (p < 0.05). The other results were compared between the SB431542 group/the U0126 group and 3.0% ISPOC group. The MFI of PI staining increased, but the expression levels of activin A, P-Smad2/3, and P-ERK1/2 decreased (p < 0.05). The expression level of ERK1/2 protein in all groups exhibited no change (p > 0.05).ConclusionResults of this study showed that 3.0% concentration of isoflurane postconditioning provided better neuroprotection than 1.5% and 4.5% concentrations of isoflurane. Activin A/Smad 2/3 and activin A/ERK1/2 signaling pathway may be involved in ISPOC-induced neuroprotection.     

Alterations of Na+/K+-ATPase,cholinergic and antioxidant enzymes activity by protocatechuic acid in cadmium-induced neurotoxicity and oxidative stress in Wistar rats

BackgroundThis study assessed the possible protective mechanisms of protocatechuic acid (PCA) against cadmium (Cd)-induced oxidative stress and neurotoxicity in rats.MethodsMale wistar strain rats weighing between 150–160 g were purchased and acclimatized for two weeks. The rats were divided into seven groups of seven each; NC group received normal saline, CAD group received 6 mg/kg of Cd-solution, CAD + PSG group received Cd-solution and prostigmine (5 mg/kg), CAD + PCA-10 and CAD + PCA-20 groups received Cd-solution and PCA (10 mg/kg and 20 mg/kg) respectively, PCA-10 and PCA-20 groups received 10 mg/kg and 20 mg/kg PCA each. Animals were administered normal saline, Cd and PCA daily by oral gavage for 21 days. After which the animals were sacrificed, the brain excised, homogenized and centrifuged. The activities of enzymes (Na⁺/K⁺-ATPase, cholinesterases, catalase, glutathione peroxidase, superoxide dismutase) and levels of oxidative stress markers (lipid peroxidation and reduced glutathione) linked to neurodegeneration were subsequently assessed.ResultsSignificant (p < 0.05) alterations in the enzyme activities and levels of oxidative stress markers were observed in CAD group when compared to the NC group. However, the activities of the enzymes were reversed in CAD + PSG and CAD + PCA groups.ConclusionsPCA may protect against cadmium-induced neurotoxicity by altering the activities of Na⁺/K⁺-ATPase, acetylcholinesterase, butyrylcholinesterase and endogenous antioxidant enzymes.     

Early administration of xenon or isoflurane may not improve functional outcome and cerebral alterations in a porcine model of cardiac arrest

BackgroundXenon (Xe) is neuroprotective when given 1 h after cardiopulmonary resuscitation (CPR). Here, we investigated if an earlier administration of Xe or isoflurane (Iso) would also reduce neurological dysfunction.Methods10 min after CPR from 8 min of cardiac arrest 21 pigs were randomized to three groups (n = 7/group) and then ventilated for 1 h with gas mixtures as follows: (1) control: 30% O₂ + 70% N₂; (2) Iso: 30% O₂ + 69% N₂ + 1% Iso; (3) Xe: 30% O₂ + 70% Xe. Physiological variables were obtained before cardiac arrest and 10, 60 and 240 min post-CPR including cardiac output (CO) and mean arterial pressure (MAP). Four days after CPR we assessed functional performance using an established neurocognitive test and overall neurological status using a neurologic deficit score (NDS). On day 5, brains of the re-anaesthetized pigs were harvested for neurohistopathological analyses.ResultsPrior to CPR there were no differences in hemodynamics and neurological status between groups. CO and MAP were significantly reduced after starting Iso administration. Both variables were also significantly lower in comparison to Xe and control animals. Control animals presented severe neurological dysfunction as measured by the NDS and the neurocognitive tests. Although Xe and Iso animals showed slightly better functional outcome this trend was not significant. Histopathological evaluation revealed ischaemic damage of neurons predominantly in the CA1 sector of the hippocampus with no differences between groups.ConclusionsIn this study early administration of Xe and Iso did not significantly reduce neurological dysfunction and histopathological alterations induced by cardiac arrest and CPR.     

Physical activity and inactivity among different body composition phenotypes in individuals with moderate to very severe chronic obstructive pulmonary disease

BackgroundThe phenotype profiling of individuals with chronic obstructive pulmonary disease (COPD) according to impairments in body composition and level of physical activity in daily life (PADL) needs to be determined.ObjectiveTo verify if individuals with COPD classified as physically active/inactive present different characteristics within different body composition phenotypes.MethodsIndividuals with COPD were cross-sectionally stratified into four groups according to fat-free and fat mass indexes: Normal Body Composition (NBC), Obese (Ob), Sarcopenic (Sarc), and Sarcopenic/Obese (Sarc/Ob). Additionally, individuals had their PADL level objectively assessed through activity monitoring during two weekdays for at least 10 h/day, and then were classified as physically active (Act) or inactive (Inact) according to international recommendations. Lung function (spirometry), exercise capacity (6-minute walking test [6MWT]) and peripheral muscle strength (1-repetition maximum [1RM]) were also assessed.Results176 individuals with COPD (mean ± standard deviation age: 67 ± 8 years, body mass index 26 ± 6 kg/m², FEV1 47 ± 16%predicted) were classified as: NBC + Act (17%), NBC + Inact (22%), Ob + Act (6%), Ob + Inact (10%), Sarc + Act (12%), Sarc + Inact (9%), Sarc/Ob + Act (8%) and Sarc/Ob + Inact (16%). The Sarc/Ob + Inact group presented lower 6MWT and 1RM for knee extension compared to NBC + Act, NBC + Inact, and Ob + Act groups (p < 0.05). The Sarc/Ob + Inact group also presented lower FEV1% predicted, 1RM for elbow flexion and elbow extension compared to the NBC + Act and NBC + Inact groups and lower 1RM for elbow extension compared to Ob + Inact group (p < 0.05).ConclusionThe combination of sarcopenia, obesity, and physical inactivity was shown to be detrimental in individuals with COPD. Therefore, this profile is a main therapeutic target for improving PADL level and/or body composition.     

Transcranial direct current stimulation as an adjunct to cognitive training for older adults with mild cognitive impairment: A randomized controlled trial

BackgroundCognitive training (CT) for individuals with mild cognitive impairment (MCI) may not be optimal for enhancing cognitive functioning. Coupling CT with transcranial direct current stimulation (tDCS) may maximize the strength of transmission across synaptic circuits in pathways that are stimulated by CT. The synergistic effects arising from this combination could be superior to those with administration of CT alone.ObjectivesTo investigate whether the receiving tDCS combined with CT is superior to CT alone on domain-specific and task-specific cognitive outcomes in older adults with MCI.MethodsThis double-blind, sham-controlled randomized trial included 67 older adults with MCI assigned to 3 groups: 1) tDCS combined with CT (tDCS + CT), 2) sham tDCS combined with CT (sham tDCS + CT) and 3) CT alone. Nine sessions of computerized CT were administered to the 3 groups for 3 weeks. In addition, tDCS and sham tDCS was delivered to the left dorsolateral prefrontal cortex to the tDCS + CT and sham tDCS + CT groups, respectively, simultaneously with CT. Standardized cognitive assessments were performed at baseline, post-intervention, and at 6-week follow-up. Participants’ performance in the CT tasks was rated every session.ResultsThe 3 groups showed improvements in global cognition and everyday memory (P < 0.017) after the intervention and at follow-up, with larger effect sizes in the tDCS + CT than other groups (d > 0.94) but with no significant differences between groups. Regarding CT outcomes, the groups showed significant differences in favour of the tDCS + CT group in decreasing the completion and reaction times of working memory and attention activities (P < 0.017).ConclusionstDCS combined with CT was not superior to sham tDCS with CT and CT alone in its effects on domain-specific cognitive outcomes, but it did provide comparatively larger effect sizes and improve the processing speed of task-specific outcomes.ClinicalTrials.govNCT03441152.     

Bidis — hand-rolled,Indian cigarettes: Induced biochemical changes in plasma and red cell membranes of human male volunteers

ObjectiveTo investigate the effect of bidi smoking on erythrocyte antioxidant status, membrane fluidity.Design and methodsThirty experimental and control subjects (mean age 35 ± 5) were selected for the study. Experimental subjects smoke 22 ± 4 bidis per day for 8–10 years.ResultsIncrease in plasma total cholesterol, LDL-cholesterol, triglycerides, lipid peroxidation, protein carbonyls with a decrease in HDL-cholesterol, thiol groups as well as increased erythrocyte catalase (CAT), superoxide dismutase (SOD), decreased glutathione peroxidase (GPx) activity and reduced glutathione (GSH) content was observed in bidi smokers. Increase in the erythrocyte membrane lipid peroxidation, cholesterol phospholipids (C/P) ratio as well as decrease in protein and Na⁺/K⁺-ATPase activity was observed. Increase in nitrite/nitrate (NOx) levels of plasma, red cell lysate was positively correlated with C/P ratio (r = 0.614) and Na⁺/K⁺-ATPase (r = 0.435) in bidi smokers.ConclusionsBidi smoke alters antioxidant status, red cell membrane fluidity and increases atherogenicity.     

A novel and automated assay for thiol/disulphide homeostasis

ObjectivesTo develop a novel and automated assay determining plasma thiol/disulphide homeostasis, which consists of thiol–disulphide exchanges.Design and methodsNative thiol and total thiol concentrations were synchronously measured as a paired test. In the first vessel, the amount of native thiol groups was measured by a modified Ellman reagent. At the parallel run, first, dynamic disulphide bonds were reduced to free thiol groups by NaBH₄. The unused reductant remnants were completely removed by formaldehyde. Thus, the total thiol amount could be accurately measured. Mercaptoethanol solutions were used as calibrators. The half value of the difference between total thiol and native thiol amounts gave the disulphide bond amount.ResultsNo separation step for the assay was needed. All processes were performed using an automated analyser within about 10 min. Plasma disulphide levels were 17.29 ± 5.32 μmol/L, native thiol levels were 397 ± 62 μmol/L and disulphide/native thiol per cent ratios were 4.32 ± 1.49 in healthy subjects. Plasma disulphide levels were higher in patients with degenerative diseases and lower in patients with proliferative diseases.ConclusionAn easy, inexpensive, practical, fully automated and also optionally manual spectrophotometric assay can be used to determine plasma dynamic thiol/disulphide homeostasis.     

Cerebral energy failure following experimental cardiac arrest: Hypothermia treatment reduces secondary lactate/pyruvate-ratio increase

AimThis was an experimental study performed to investigate cerebral metabolism during hypothermia treatment and rewarming after resuscitation from cardiac arrest (CA).Materials and methodsSixteen pigs underwent CA followed by cardiopulmonary resuscitation (CPR). After randomisation into one hypothermic (n = 8) and one normothermic group (n = 8) the animals received infusion of 4 or 38 °C saline, respectively. Following restoration of spontaneous circulation (ROSC) both groups were observed for 360 min. The hypothermic group was cooled for 180 min and then rewarmed. Temperature was not modulated in the normothermic group.Cerebral microdialysis was conducted and lactate/pyruvate (L/P)-ratio and glutamate were analysed. Intracranial pressure probe was inserted. Oxygen saturation in venous jugular bulb blood (SjO₂) was analysed.ResultsAll animals initially had increased L/P-ratio (>30). A total of nine animals developed secondary increase. In the hypothermic group this was observed in 2/7 animals and in the normothermic group in 7/8 (p = 0.04). Glutamate increased initially in all animals with secondary increases in two animals in each group. No differences in L/P-ratio or glutamate were detected during the rewarming phase compared to the hypothermic phase. The hypothermic group had higher SjO₂ (p = 0.04). In both groups intracranial pressure increased after ROSC.ConclusionAfter resuscitation from CA there was a risk of cerebral secondary energy failure (reflected as an increased L/P-ratio) but hypothermia treatment seemed to counteract this effect. Cerebral oxygen extraction, measured by SjO_2, was increased in the hypothermic group probably due to reduced metabolism. Rewarming did not reveal any obvious harmful events.     

Ghrelin protects against palmitic acid or lipopolysaccharide-induced hepatocyte apoptosis through inhibition of MAPKs/iNOS and restoration of Akt/eNOS pathways

BacgroundGhrelin has been shown to exert various biological functions. However, the effect and mechanism of ghrelin on PA- or LPS-induced liver injury remains unknown.MethodsNormal human hepatocyte lines (LO2 and 7701) were pretreated with ghrelin (10⁻⁸ M) for 30 min before stimulation with lipopolysaccharide (LPS) or palmitic acid (PA). The proliferation and apoptosis of cells were detected with CCK8, Hoechst staining and flow cytometric analysis. Levels of NO of cell supernatants were examined by enzyme-linked immunosorbent assay (ELISA). The protein levels and mRNA of target genes of endothelial NOS (eNOS) and inducible NOS (iNOS) were measured by western blotting, immunofluorescence and quantitative real-time polymerase chain reaction (qRT-PCR). The expression of Bax, Bcl2, caspase 3, p-Akt, p-P38 and p-JNK were detected by western blotting.ResultsResults of CCK8, Hoechst staining and flow cytometric analysis showed that ghrelin-pretreatment attenuated LPS- or PA- induced cellular proliferation inhibition and apoptosis induction. ELISA results revealed that ghrelin pretreatment reduced levels of NO of cell supernatants (P < 0.05). Results of western blotting and immunofluorescence showed that protein levels of iNOS in ghrelin- pretreated group were significantly reduced compared with LPS- or PA- treated group, while protein levels of eNOS were restored by ghrelin pretreatment. Results of qRT-PCR showed that mRNA levels of Bax, iNOS were reduced by ghrelin pretreatment, while levels of mRNA of Bcl2 and eNOS were increased (P < 0.05). The protein levels of pAkt were significantly increased by ghrelin pretreatment, while the protein levels of p-JNK, p-P38 and caspase 3 were reduced. The restoration of eNOS could be reversed by an Akt inhibitor.ConclusionsGhrelin pretreatment attenuated LPS- or PA-induced hepatocyte apoptosis, which may least partly via inhibition of mitogen-activated protein kinases (MAPKs)/iNOS and restoration of Akt/eNOS pathways.     

FK228 augmented temozolomide sensitivity in human glioma cells by blocking PI3K/AKT/mTOR signal pathways

Temozolomide is a novel cytotoxic agent currently used as first-line chemotherapy for glioblastoma multiforme (GBM). Romidepsin (FK228), a histone deacetylase inhibitor, is a promising new class of antineoplastic agent with the capacity to induce growth arrest and/or apoptosis of cancer cells. However, combination of the two drugs in glioma remains largely unknown. In the present study, we evaluated the combinatory effects of FK228 with TMZ in glioma, and its molecular mechanisms responsible for these effects. Glioma cell lines were treated with TMZ, FK228 or the combination of drugs. The resistance effect including cytotoxicity and apoptosis was determined in glioma cells, respectively. We further evaluated the effects of FK228 in the PI3K/Akt-signaling pathway in vitro. Mice engrafted with 5 × 10⁶ LN382 cells were treated with TMZ, FK228 or the combination of two drugs, and tumor weights and volumes were measured, respectively. FK228 enhanced the cytotoxic effects of TMZ in glioma cells compared to vehicle-treated controls or each drug alone. The combination of FK228 and TMZ-induced apoptosis was demonstrated by increased expression of cleaved-Caspase 3, Bax, cleaved-PARP, and decreased Bcl-2 expression. Furthermore, the expression of key components of the PI3K/Akt-signaling pathway showed that combination of FK228 and TMZ block PI3K/Akt pathways in vitro. This block effect was also confirmed in vivo in mice models. Mice treated with both FK228 and TMZ drugs showed significantly reduced tumor weights and volumes, compared to each drug alone. Our results suggested that FK228 augmented temozolomide sensitivity in human glioma cells partially by blocking PI3K/AKT/mTOR signal pathways. It thus may provide a promising target for improving the therapeutic outcome of TMZ-resistant gliomas, although further studies will be needed.     

A 4-arm randomized controlled pilot trial of innovative solutions for jugular central venous access device securement in 221 cardiac surgical patients

PurposeTo improve jugular central venous access device (CVAD) securement, prevent CVAD failure (composite: dislodgement, occlusion, breakage, local or bloodstream infection), and assess subsequent trial feasibility.Materials and MethodsStudy design was a 4-arm, parallel, randomized, controlled, nonblinded, pilot trial. Patients received CVAD securement with (i) suture + bordered polyurethane (suture + BPU; control), (ii) suture + absorbent dressing (suture + AD), (iii) sutureless securement device + simple polyurethane (SSD + SPU), or (iv) tissue adhesive + simple polyurethane (TA + SPU). Midtrial, due to safety, the TA + SPU intervention was replaced with a suture + TA + SPU group.ResultsA total of 221 patients were randomized with 2 postrandomization exclusions. Central venous access device failure was as follows: suture + BPU controls, 2 (4%) of 55 (0.52/1000 hours); suture + AD, 1 (2%) of 56 (0.26/1000 hours, P = .560); SSD + SPU, 4 (7%) of 55 (1.04/1000 hours, P = .417); TA + SPU, 4 (17%) of 23 (2.53/1000 hours, P = .049); and suture + TA + SPU, 0 (0%) of 30 (P = .263; intention-to-treat, log-rank tests). Central venous access device failure was predicted (P < .05) by baseline poor/fair skin integrity (hazard ratio, 9.8; 95% confidence interval, 1.2-79.9) or impaired mental state at CVAD removal (hazard ratio, 14.2; 95% confidence interval, 3.0-68.4).ConclusionsJugular CVAD securement is challenging in postcardiac surgical patients who are coagulopathic and mobilized early. TA + SPU was ineffective for CVAD securement and is not recommended. Suture + TA + SPU appeared promising, with zero CVAD failure observed. Future trials should resolve uncertainty about the comparative effect of suture + TA + SPU, suture + AD, and SSD + SPU vs suture + BPU.     

Effect of self-tailored high-intensity interval training versus moderate-intensity continuous exercise on cardiorespiratory fitness after myocardial infarction: A randomised controlled trial

BackgroundWhether high-intensity interval training (HIIT) is more efficient than moderate-intensity continuous exercise (MICE) to increase cardiorespiratory fitness in patients with acute coronary syndrome at moderate-to-high cardiovascular risk is controversial. The best approach to guide training intensity remains to be determined.ObjectiveWe aimed to assess intensities achieved with self-tailored HIIT and MICE according to perceived exertion and to compare the effect on cardiorespiratory fitness in patients early after ST-elevation myocardial infarction (STEMI).MethodsWe included 69 males starting cardiac rehabilitation within 4 weeks after STEMI. After a 3-week run-in phase with MICE, 35 patients were randomised to 9 weeks of HIIT (2 × HIIT and 1 × MICE per week) and 34 patients to MICE (3 × MICE). Training workload for MICE was initially set at the patients’ first ventilatory threshold (VT). HIIT consisted of 4 × 4-min intervals with a workload above the second VT in high intervals. Training intensity was adjusted weekly to maintain the perceived exertion (Borg score 13–14 for MICE, ≥ 15 for HIIT). Session duration was 38 min in both groups. Peak oxygen consumption (VO₂) was measured by cardiopulmonary exercise testing pre- and post-intervention.ResultsBoth groups improved peak VO₂ (ml/kg/min) (HIIT +1.9, P < 0.001; MICE +3.2, P < 0.001, Cohen's d ?0.4), but changes in VO₂ were not significantly different between groups (P = 0.104). Exercise regimes did not differ between groups in terms of energy expenditure or training time, but perceived exertion was higher with HIIT.ConclusionsSelf-tailored HIIT was feasible in patients early after STEMI. It was more strenuous but not superior nor more time-efficient than MICE in improving peak VO₂.The trial was registered at ClinicalTrial.gov (NCT02627586).     

The efficacy and safety of antithrombin and recombinant human thrombomodulin combination therapy in patients with severe sepsis and disseminated intravascular coagulation

PurposeRecombinant human thrombomodulin (rhTM) is often used concomitantly with antithrombin (AT) to treat disseminated intravascular coagulation (DIC). This observational study aimed to investigate the efficacy and safety of AT + rhTM combination therapy.Materials and methodsOne hundred twenty-nine patients with severe sepsis and DIC participated in this study. Of these, 78 patients were treated with AT + rhTM (AT + rhTM group) and 51 patients were treated with AT alone (AT group). We compared coagulation and inflammation markers, Sequential Organ Failure Assessment score, and DIC score at day 0 (baseline) and day 7 between the 2 groups. Bleeding events and 28-day mortality were also compared.ResultsPlatelet counts and D-dimer levels at day 7 significantly improved in the AT + rhTM group compared with the AT group, and 28-day mortality was significantly lower in the AT + rhTM group than in the AT group (AT + rhTM: 15.4% vs AT: 29.4%). During the study period, the incidence of bleeding complications was similar in both groups (AT + rhTM: 6.4% vs AT: 7.8%).ConclusionsCompared with AT monotherapy, combination therapy with AT and rhTM may be more effective in improving platelet counts and D-dimer levels, as well as reducing mortality, in patients with severe sepsis-associated DIC.     

The effect of aloe vera on ischemia—Reperfusion injury of sciatic nerve in rats

PurposeAloe vera is compound which has strong antioxidant and anti-inflammatory effects. We investigated the neuroprotective role of aloe vera treatment in rats with experimental sciatic nerve ischemia/reperfusion injury.MethodsTwenty-eight male Wistar Albino rats were divided equally into 4 groups. Groups; Control group (no surgical procedure or medication), sciatic nerve ischemia/reperfusion group, sciatic nerve ischemia/reperfusion + aloe vera group and sciatic nerve ischemia/reperfusion + methylprednisolone group. Ischemia was performed by clamping the infrarenal abdominal aorta. 24 hours after ischemia, all animals were sacrificed. Sciatic nerve tissues were also examined histopathologically and biochemically.ResultsIschemic fiber degeneration significantly decreased in the pre-treated with aloe vera and treated with methylprednisolone groups, especially in the pre-treated with aloe vera group, compared to the sciatic nerve ischemia/reperfusion group (p < 0.05). A significant decrease in MDA, an increase in NRF1 level and SOD activity were observed in the groups which obtained from the AV and MP groups when compared to the sciatic nerve ischemia/reperfusion group. When all results were analysed it was seen that the aloe vera group was not statistically different compared to the MP group (p > 0.05).ConclusionsAloe vera is effective neuroprotective against sciatic nerve ischemia/reperfusion injury via antioxidant and anti-inflammatory properties. Also aloe vera was found to be as effective as MP.     

Lactobacillus rhamnosus induced epithelial cell apoptosis,ameliorates inflammation and prevents colon cancer development in an animal model

Background/AimProbiotics have been suggested as prophylactic measure in colon carcinogenesis. This study aimed at determining the potential prophylactic activity of Lactobacillus rhamnosus GG CGMCC 1.2134 (LGG) strain on colorectal carcinogenesis via measuring its effect on Nuclear factor kappa B (NFκB) inflammatory pathway and apoptosis.Materials and methods64 Sprague Dawley rats were grouped into four as follows; Group 1 (Healthy control), Group 2 (LGG), Group 3 (cancer control Dimethyl hydrazine (DMH)) and Group 4 (LGG + DMH). LGG was administered orally to LGG and LGG + DMH groups. Colon carcinogenesis was chemically induced in LGG + DMH and DMH groups by weekly injection of 40 mg/kg DMH. Animals were sacrificed after 25 weeks of experiment and tumor characteristics assessed. The change in expression of NFκB-p65, COX-2, TNFα, Bcl-2, Bax, iNOS, VEGFα, β-catenin, Casp3 and p53 were evaluated by western blotting and qRT-PCR.ResultsLGG treatment significantly reduced tumor incidence, multiplicity and volume in LGG + DMH treatment group compared to DMH cancer control group. Also, LGG treatment reduced the expression of β-catenin and the inflammatory proteins NFκB-p65, COX-2 and TNFα; the anti-apoptotic protein Bcl-2, but increased the expression of the pro-apoptotic proteins Bax, casp3 and p53 compared with DMH group.ConclusionLGG have a potential protection effect against colon carcinogenesis; inducing apoptosis and ameliorating inflammation, and may hold a promise as bio-therapeutic dietary agent.     

Ketamine delays mortality in an experimental model of hemorrhagic shock and subsequent sepsis

BackgroundIn previous studies ketamine was reported to improve survival and decrease serum interleukin-6 (IL-6) concentration after sepsis alone and after burn injury followed by sepsis. The aim of this study was to determine whether ketamine alters survival and/or IL-6 after hemorrhagic shock alone or hemorrhagic shock followed by sepsis.Materials and methodsRats were subjected to hemorrhagic shock with or without subsequent Gram-negative bacterial sepsis and were either treated with ketamine 5 mg/kg or were not treated. Blood was sampled for IL-6 determination prior to hemorrhage, at the completion of resuscitation, and at 6 and 30 h later. Mortality was recorded for 7 days following hemorrhage or hemorrhage + sepsis.ResultsAfter hemorrhage + sepsis the time to median mortality was significantly later in the ketamine-treated group (36 h) than in the control group (12 h). At 12 h the survival rate of the ketamine-treated group (100%) was significantly higher than in the control group (55%). There were no significant differences between groups with respect to IL-6 or 7-day survival after either hemorrhage + sepsis or hemorrhage alone.ConclusionKetamine improved 12 h survival and delayed mortality after hemorrhage + sepsis without significantly altering IL-6, and did not alter survival or IL-6 after hemorrhage alone.     

Cerebral effects of three resuscitation protocols in uncontrolled haemorrhagic shock: A randomised controlled experimental study

BackgroundTo compare haemodynamic and cerebral variables during aggressive fluid resuscitation vs. administration of a hypertonic starch solution (HS) combined with either noradrenaline [norepinephrine] or arginine vasopressin in an animal model of uncontrolled haemorrhagic shock.MethodsAfter Animal Investigational Committee approval, 24 anaesthetised pigs underwent a liver trauma. At haemodynamic decompensation, animals were randomly assigned to receive fluid resuscitation (6% HES 130/0.4, 20 mL/kg, and Ringer, 40 mL/kg; FR group, n = 8), or noradrenaline (bolus 20 μg/kg, continuously 1 μg/kg/min) combined with HS (7.2% NaCl/6% HES 200/0.5; 4 mL/kg) (n = 8; NA/HS group), or vasopressin (bolus 0.2 U/kg, continuously 0.04 U/kg/min) combined with HS (4 mL/kg) (n = 8; AVP/HS group), respectively. Thirty minutes after drug administration, bleeding was controlled manually.ResultsMean arterial blood pressure (MAP), cerebral perfusion pressure (CPP), and brain tissue oxygen pressure (P_btO₂) decreased significantly with haemorrhage in all groups (p < 0.05). AVP/HS resulted in a faster and higher increase of MAP and CPP compared to both NA/HS and FR (p < 0.001 vs. FR; p < 0.01 vs. NA/HS). Compared to FR, P_btO₂ increased faster with AVP/HS and NA/HS (p < 0.05) after therapy, and ICP was lower at the end of the study period (p < 0.05). All animals (8/8) of the AVP/HS group survived, compared to 4/8 and 4/8 in the NA/HS and FR group, respectively (p = 0.07).ConclusionsFollowing uncontrolled haemorrhagic shock in this animal model, combination of HS with arginine vasopressin increased CPP and cerebral oxygenation faster than aggressive fluid resuscitation, without re-increasing ICP.     

Analytical measurement of serum 25-OH-vitamin D3, 25-OH-vitamin D2 and their C3-epimers by LC–MS/MS in infant and pediatric specimens

ObjectivesTo develop a simple and sensitive LC–MS/MS procedure for quantification of serum 25-OH-vitamin D₃ (25-OH-D₃), 25-OH-vitamin D₂ (25-OH-D₂), and their C3-epimers.MethodsSerum 25-OH-vitamin D metabolites were extracted with MTBE and quantified by LC–MS/MS. Commercially available calibrators and QC materials were employed. The ion-transition 401.2 → 365.2 was monitored for 25-OH-D₃ and C3-epi-25-OH-D₃, 407.2 → 371.3 for d₆-25-OH-D₃, 413.2 → 331.2 for 25-OH-D₂ and C3-epi-25-OH-D₂ and 419.2 → 337.1 for, d₆-25-OH-D₂. As a proof-of-principle, 25-OH-D₃ and C3-epi-25-OH-D₃ were quantified in 200 pediatric subjects (0–20 years of age). Cholecalciferol supplements were examined as a potential source of C3-epimer.ResultsThe assay provided an LLOQ of ≤ 2.8 nmol/L for all 25-OH-D metabolites, with a linear response up to 400 nmol/L. The CV was < 10% for 25-OH-D_2/3 and < 15% for C3-epi-25-OH-D₃. C3-epi-25-OH-D₃ was quantified in all subjects, with higher concentrations observed in infants ≤ 1 year of age (11.44 nmol/L vs. 4.4 nmol/L; p < 0.001). Within the first year of life, 25-OH-D₃ concentrations increased linearly, while C3-epi-25-OH-D₃ concentrations remained constant. At 12 months of age, C3-epi-25-OH-D₃ concentration dropped by almost 50% (11.4 nmol/L in infants ≤ 1 year of age vs. 5.4 nmol/L in infants 1–2 years of age; p < 0.001). Liquid vitamin D₃ supplements did not contain appreciable amounts of C3-epi-D₃.ConclusionsThe proposed LC–MS/MS procedure is suitable for quantifying 25-OH-D₃ metabolites. Although the C3-epimer is present in all pediatric subjects, it is significantly elevated in individuals ≤ 1 year of age and drops at 12 months of age. Oral vitamin D supplements are unlikely to be a significant source of C3-vitamin D epimer.     

Dysregulation of TH type cytokines in the patients of Parthenium induced contact dermatitis

BackgroundParthenium contact dermatitis is a major health problem caused by a cosmopolitan weed Parthenium hysterophorus. It is a T cell-mediated immune injury and disease manifests as itchy erythematous papules, papulovesicular and plaque lesions on exposed areas of the body. We studied the involvement of T_H1/T_H2/T_H17/Treg type responses by assaying various cytokines in Parthenium dermatitis.MethodsThe study includes 50 patients of Parthenium dermatitis confirmed by patch testing and 50 healthy subjects. The serum levels of T_H1, T_H2, T_H17 and Treg cytokines were estimated by high sensitivity sandwich ELISA and were compared statistically between groups using ANOVA.ResultsThe mean concentration of T_H1 cytokines (p < 0.001) and IL-17 (p < 0.001) were increased significantly as compared to controls. In contrast, decrease in levels of IL-10 (p < 0.002) and TGF-β (p < 0.001) were significant and levels of IL-4 (p < 0.262) were insignificant whereas no alterations in the total IgE concentrations (p < 0.976) was observed.ConclusionThe induction of T_H1 and T_H17 cytokines reinforce the need of detailed analysis of immune dysregulation in Parthenium dermatitis and might add some insight in the pathogenesis, diagnosis and current treatment modalities of this disease.