Comparative evaluation of two dye probes in the rat everted gut sac model for unambiguous classification of P-gp substrate and inhibitor

IntroductionP-glycoprotein (P-gp) plays a crucial role in beta-amyloid efflux from the blood–brain barrier thus becoming a promising pharmacological target in the treatment of Alzheimer's disease (AD). The increase of P-glycoprotein expression and activity by a P-gp inducer could be an effective pharmacological strategy in slowing or halting the progression of AD. Commonly used in vitro methods to classify a P-gp interacting molecule as substrate, inhibitor, modulator or inducer are not always confirmed by in vivo experiments. Here we validate the new dye-probe beta-amyloid (1–40) HiLyte Fluor? TR-labeled (Ab-HiLyte) (Anaspec) P-gp mediated transport in the ex vivo rat everted gut sac assay by using MC18 or MC266, a fully characterized P-gp inhibitor and substrate, respectively, and compare it with the commonly used dye rhodamine.MethodsMale Wistar rats' everted intestines were divided into sacs, each sac was filled with 10 μM Ab-HiLyte with or without 50 μM of MC18 or MC266. Ab-HiLyte concentrations in mucosal fluid were measured spectrophotometrically at 594 nm at each appropriate time.ResultsThe Ab-HiLyte P-gp mediated efflux had a K = 1.00 × 10^? 2 min^? 1 and t_1/2 = 68.74 min, while in the presence of MC18, the Ab-HiLyte efflux turned out to be reduced by an order of magnitude (K = 1.65 × 10^? 3 min^? 1) and the half life is extremely increased (t_1/2 = 419 min). A P-gp substrate, like MC266, determines no change in the efflux of Ab: the kinetic constant and the half life turned out to be unmodified (K = 1.81 × 10^? 2 min^? 1 and t_1/2 = 38.28 min).DiscussionThe results demonstrate that the new dye probe, Ab-HiLyte, could be a probe of choice to unequivocally distinguish between a P-gp substrate and an inhibitor. This is particularly important as different groups obtain a controversial classification of the same compound.     

Gabapentin-induced changes of plasma cortisol level and immune status in hysterectomized women

AimWe have examined the effects of gabapentin (GBP) on stress-related changes of cortisol and catecholamines in patients who underwent hysterectomy because of uterine fibrinoids. Additionally, we have observed the effect of GBP on the immune status in the acute stress response to surgery.MethodsSixty patients scheduled for an abdominal hysterectomy were randomly assigned to the GBP administration 1 h before surgery (n = 30 pts), or to the placebo group (n = 30 pts). Blood samples were collected before and 24 h after the surgery. The intensity of pain was assessed by a visual analogue scale (VAS) every 8 h at rest. Immunomodulatory effects of GBP were determined by flow cytometry. We followed the total proportion of CD3⁺ lymphocytes, CD3⁺CD4⁺, CD3⁺CD8⁺, CD19⁺ B lymphocytes, CD16⁺CD56⁺CD3⁻NK cells and CD16⁺CD56⁺CD3⁺ NKT cells before and 24 h after hysterectomy. The plasma cortisol and catecholamines concentration was used to estimate the level of the stress response.ResultsVAS pain score at rest was significantly lower in the GBP group than in the placebo group (P = 0.003). Application of GBP significantly decreased the plasma cortisol level 24 h after the operation in comparison to the placebo group (P < 0,001). We found significant positive correlation between the VAS pain score and concentration of cortisol in all patients (P = 0.025). GBP reduced the concentration of catecholamines (p < 0.05). The proportion of CD3⁺ (P = 0.027) and CD3⁺CD4⁺cells (P = 0.006) was significantly lower in the GBP group 24 h after operation, while the contribution of CD19⁺ (P = 0.033) was significantly higher.ConclusionPreoperative administration of GBP reduced the pain scores at rest in patients at 0, 16 and 24 h after abdominal hysterectomy. Additionally, GBP reduced the stress response and changed immune parameters in the reaction to surgery.     

Monitorización de la vacomicina en administración intraperitoneal en pacientes sometidos a diálisis peritoneal continua ambulatoria

ObjectiveTo validate a pharmacokinetic model of the treatments with intraperitoneal vancomycin applied to patients on continuous ambulatory peritoneal dialysis with bacterial peritonitis.MethodsTo carry out a prospective study divided in 2 cohorts: the first one including ten patients of 56 ± 14 years and 65 ± 5 kg, and the second one with 10 patients (12 episodes of peritonitis) aged 52 ± 13 years and 64 ± 8 kg. The treatment consists of administering and retaining for 6 h in the peritoneal cavity a solution containing 2 g of vancomycin and 1 g of ceftazidime into 2 l of “dialysis solution”. After the antibiotic administration, blood samples were obtained at 4, 6, 8, 10, 24, 48 and 168 h in the first cohort and at 6 and 120 h (C^VAN₁₂₀) in the second. The pharmacokinetic model was developed from the parameters obtained from the first cohort and was validated by the second cohort, calculating the mean error (ME) and the mean squared prediction error (MSPE) of the C^VAN₁₂₀.ResultsVancomycin serum concentrations fell from 39.63 ± 7.62 mcg/ml at 4 h to 8.55 ± 2.87 mcg/ml at 168 h for the first cohort, and from 37.65 ± 6.84 mcg/ml at 6 h to 10.82 ± 2.66 mcg/ml at 120 h (C^VAN₁₂₀) for the second cohort. The pharmacokinetics parameters were: C1 = 0.006 1/h/kg and Vd: = 0.52 1/kg for the first cohort, and C1 = 0.006 1/h/kg and Vd: = 0.53 1/kg for the second. The predictive ME and MSPE of the C^VAN₁₂₀. were 0.59 mcg/ml ([EM*100/C^VAN₁₂₀ = 5.5%) and 10.38 mcg²/ml² ([MES*100/(C^VAN₁₂₀)²]) respectively.ConclusionThe presented model shows an adequate exactitude and precision for the monitoring of intraperitoneal vancomycin in patients submitted to continuous ambulatory peritoneal dialysis with peritonitis.     

A clinical comparison of the efficacy and safety of biosimilar G-CSF and originator G-CSF in haematopoietic stem cell mobilization

BackgroundRecombinant granulocyte colony-stimulating factor (G-CSF) is widely used to mobilize haematopoietic stem cells. We compared the efficacy and safety of a biosimilar G-CSF (Zarzio^®, Sandoz Biopharmaceuticals) with the originator G-CSF (Neupogen^®, Amgen) in patients with haematological malignancies.MethodsA total of 108 patients were included in this study, 59 of whom were female (49 male), with an overall median age of 51 years (range 19–69). Patients had multiple myeloma (n = 46), non-Hodgkin's lymphoma (n = 28), Hodgkin's lymphoma (n = 26), or other diagnosis (n = 8). After administration of mobilizing regimens (primarily high-dose etoposide, high-dose cyclophosphamide, intermediate-dose Ara-C or ESHAP), patients were randomized to a standard daily 10 μg/kg dose of biosimilar G-CSF (n = 54) or originator G-CSF (n = 54).ResultsMedian duration of G-CSF administration was 8 days with both biosimilar G-CSF (range 4–17) and originator G-CSF (range 4–14). Both groups had a median of one apheresis with a median time until first apheresis of 11 days. There were no statistically significant differences between groups in the mean ± SD number of mobilized CD34+ cells/μL in peripheral blood or the number of CD34+ cells/kg body weight. Five patients (9%) in the originator G-CSF group and six patients in the biosimilar G-CSF group (11%) did not mobilize sufficient CD34+ cells. The adverse event profile was similar between groups.ConclusionsA biosimilar G-CSF (Zarzio^®) demonstrated similar efficacy and safety as the reference originator G-CSF (Neupogen^®) in hematopoietic stem cell mobilization in patients with haematological malignancies.     

Determination of lamivudine and zidovudine permeability using a different ex vivo method in Franz cells

IntroductionThe major processes that control the absorption of orally administered drugs are dissolution and gastrointestinal permeation. These processes depend on two main properties: solubility and permeability. Based on these characteristics, the Biopharmaceutical Classification System (BCS) was proposed as a tool to assist in biowaiver and bioavailability prediction of drugs.MethodsThe purpose of the present study was to evaluate the permeability of lamivudine (3TC) and zidovudine (AZT) using a different ex vivo method in Franz cells. A segment of jejunum was inserted in a Franz cells apparatus, in order to assess drug permeability in the apical–basolateral (A–B) and basolateral–apical (B–A) directions. Each drug was added to the donor chamber, collected from the acceptor chamber and analyzed by HPLC. Fluorescein (FLU) and metoprolol (METO) were used as low and high permeability markers, respectively.ResultsThe apparent permeability (P_app) results for the A–B direction were: P_{app FLU A–B} = 0.54 × 10^? 4 cm·s^? 1, P_{app METO A–B} = 7.99 × 10^? 4 cm·s^? 1, P_{app 3TC A–B} = 4.58 × 10^? 4 cm·s^? 1 and P_{app AZT A–B} = 5.34 × 10^? 4 cm·s^? 1. For the B–A direction, the P_app results were: P_{app FLU B–A} = 0.56 × 10^? 4 cm·s^? 1, P_{app METO B–A} = 0.25 × 10^? 4 cm·s^? 1, P_{app 3TC B–A} = 0.24 × 10^? 4 cm·s^? 1 and P_{app AZT B–A} = 0.19 × 10^? 4 cm·s^? 1.DiscussionFor the A–B direction, the P_app results of fluorescein and metoprolol show low and high permeability, respectively, indicating that the membranes were appropriate for permeability studies. For the A–B direction, the P_app results of 3TC and AZT suggest that these antiretroviral drugs have permeability values close to metoprolol. Nevertheless, for the B–A direction the P_app results do not suggest efflux mechanism for any of the drugs. Thereby, the different ex vivo methods using Franz cells can be successfully applied in drug permeability studies, in particular for drug biopharmaceutical classification.     

Associations between PTSD and intimate partner and non-partner aggression among substance using veterans in specialty mental health

BackgroundRisk factors of violence perpetration in veterans include substance use and posttraumatic stress disorder (PTSD); however, it is unknown whether these factors are associated with greater risk for partner or non-partner violence. This study investigated the associations between probable PTSD, heavy drinking, marijuana use, cocaine use, and partner and non-partner violence perpetration.MethodsSelf-report questionnaires assessing past-year partner and non-partner aggression (CTS2) as well as past-month substance use (SAOM), probable PTSD (PCL-C), and probable depression (PHQ-9) were administered to 810 substance using veterans entering VA mental health treatment.ResultsIn bivariate analyses, probable PTSD in substance using veterans was associated with violence perpetration (partner physical, χ² = 11.46, p = 0.001, φ = 0.12; non-partner physical, χ² = 50.64, p < 0.001, φ = 0.25; partner injury, χ² = 6.41, p = 0.011, φ = 0.09; non-partner injury, χ² = 42.71, p < 0.001, φ = 0.23). In multiple logistic regression analyses that adjusted for sociodemographic characteristics, probable PTSD was independently associated with non-partner physical (odds ratio [OR], 2.82; 95% confidence interval [CI], 1.97–4.05) and injury aggression (OR, 3.96; CI, 2.56–6.13). Cocaine and heavy drinking were independently associated with non-partner physical and injury aggression and non-partner injury aggression respectively.ConclusionsThe results provide evidence that probable PTSD, heavy drinking, and cocaine use are associated with increased risk of non-partner violence perpetration in substance using veterans. These results underscore the importance of screening for PTSD symptoms and violence perpetration towards non-partners in substance using veterans presenting for treatment.     

Binding of new aminopropan-2-ol compounds to bovine serum albumin, α1 acid glycoprotein and rat serum using equilibrium dialysis and LC/MS/MS

BackgroundThe binding of three new aminopropan-2-ol compounds briefly called 2F109, ANBL and TWo8 with potential cardiovascular activity to bovine serum albumin (BSA), α₁-acid glycoprotein (AGP) and to rat serum was studied. The chemical structures of these compounds are related to carvedilol. They possess an antiarrhythmic and hypotensive activity, and β- and α-adrenolytic mechanism of action. All analogues are weak bases with pKa values 8.65,8.85 and 8.26 for 2F109, ANBL and TWo8, respectively, and they possess lipophilic character (log P > 1.9584).MethodsThe extent of protein binding was determined using equilibrium dialysis in the range 2.5 – 900 μM, and 2.5 – 300 μM for binding of investigated compounds to BSA and AGP, respectively, and the quantitative measurement was done by LC/ESI-MS/MS assay.ResultsThe studied compounds bound to a single class of binding sites on BSA which was characterized by low affinity (K_d for 2F109 = 8.49 × 10^–5 M, for ANBL = 1.92 × 10^–5 M, and for TWo8 = 1.71 × 10^–5 M) and low capacity(n = 0.53 for 2F109,0.132 for ANBL and 0.13 for TWo8). The binding of 2F109, ANBL and TWo8 to AGP revealed one class of binding sites, with moderate affinity (K_d for 2F109 = 4.67 × 10^–6 M, for ANBL = 3.48 × 10^–5 M, and for TWo8 = 1.13 × 10^–5 M) and higher capacity (n = 2.21 for 2F109, 2.76 for ANBL and 2.28 for TWo8).ConclusionThe obtained data indicate that 2F109, ANBL and TWo8 moderately bind to BSA (34.2 – 71.2%) with low capacity (K_a = 6.21 × 10³–7.61 × 10³ M^–1)and strongly bind to AGP(71.5–85.5%)with moderate affinity (K_a = 7.94 × 104⁴.73 ×10⁵ M^–1).     

Effects of alcohol on disinhibition towards alcohol-related cues

BackgroundWe investigated (1) the effects of acute alcohol on inhibition of alcohol-related versus neutral cues, (2) the effects of drinking status on inhibition of alcohol-related versus neutral cues, and (3) the similarity of any effects of alcohol or drinking status across two different cue types (lexical versus pictorial).MethodsParticipants received 0.0 g/kg, 0.4 g/kg or 0.6 g/kg of alcohol in a between-subjects design. Healthy, heavy and light social alcohol users (n = 96) completed both lexical and pictorial cue versions of an alcohol-shifting task. Participants were instructed to respond to target stimuli by pressing the spacebar, but to ignore distracter stimuli. Errors towards distracter stimuli were analysed using a series of mixed-model ANOVAs, with between-subjects factors of challenge and drinking status and within-subjects factors of distracter type (alcohol, neutral) and block (shift, non-shift).ResultsLexical commission error data indicated a main effect of distracter (F [1,90] = 43.25, p < 0.001, η² = 0.33), which was qualified by a marginal interaction with challenge condition (F [2,90] = 2.77, p = 0.068, η² = 0.06). Following an acute high dose of alcohol participants made more errors towards alcohol distracters. Pictorial commission error data indicated a significant main effect of distracter (F [1,90] = 67.40, p < 0.001, η² = 0.43), such that all participants made more errors towards neutral image distracters versus alcohol distracter images.ConclusionsOur results reveal acute alcohol's impairment of inhibitory control may be enhanced when a response towards alcohol-related lexical stimuli is required to be withheld.     

Case–control association analysis of polymorphisms in the delta-opioid receptor,OPRD1, with cocaine and opioid addicted populations

BackgroundAddiction susceptibility and treatment responsiveness are greatly influenced by genetic factors. Sequence variation in genes involved in the mechanisms of drug action have the potential to influence addiction risk and treatment outcome. The opioid receptor system is involved in mediating the rewarding effects of cocaine and opioids. The μ-opioid receptor (MOR) has traditionally been considered the primary target for opioid addiction. The MOR, however, interacts with and is regulated by many known MOR interacting proteins (MORIPs), including the δ-opioid receptor (DOR).MethodsThe present study evaluated the contribution of OPRD1, the gene encoding the DOR, to the risk of addiction to opioids and cocaine. The association of OPRD1 polymorphisms with both opioid addiction (OA) and cocaine addiction (CA) was analyzed in African American (OA n = 336, CA n = 503) and European American (OA n = 1007, CA n = 336) populations.ResultsThe primary finding of this study is an association of rs678849 with cocaine addiction in African Americans (allelic p = 0.0086). For replication purposes, this SNP was analyzed in a larger independent population of cocaine addicted African Americans and controls and the association was confirmed (allelic p = 4.53 × 10^?5; n = 993). By performing a meta-analysis on the expanded populations, the statistical evidence for an association was substantially increased (allelic p = 8.5 × 10^?7) (p-values non-FDR corrected).ConclusionThe present study suggests that polymorphisms in OPRD1 are relevant for cocaine addiction in the African American population and provides additional support for a broad role for OPRD1 variants in drug dependence.     

Shift from darbepoetin-α to continuous erythropoietin receptor activator decreases serum aluminium concentration in patients on hemodialysis

ObjectiveThe response of erythropoietic stimulating agents (ESA) in uremic patients may be associated with the changes of biochemical parameters, metal elements and inflammation status during the shift from one ESA to another.MethodWe compared changes in above mentioned factors after switching from darbepoetin-α (DPO) 20 μg weekly for 10 weeks to continuous erythropoietin receptor activator (CERA) 100 μg monthly for 10 weeks in uremic patients on hemodialysis. The haematocrit (Hct), metal elements and inflammation status are the primary outcome. Subjects included 54 patients without transfusion or bleeding or additional ESAs. Responders (IR, n = 36) were defined as patients with an increase in Hct after the swtich.ResultAlthough there was no significant difference in overall mean Hct after the switch (p = 0.135), there are significantly greater mean number of red blood cells (RBC) (p = 0.006), higher platelet numbers (p = 0.001), larger RBCs (p = 0.017) and higher creatinine (p = 0.04) and total cholesterol (T-CHOL) (p = 0.003) levels. Mean overall aluminium (Al) level decreased significantly (p = 0.001). C-reactive protein (CRP) also decreased (p = 0.016). The overall LDH increased (p = 0.049) and potassium decreased significantly (p = 0.036), which indicating active erythropoiesis. The calcium (Ca) level was significantly higher (p = 0.034) and phosphate was significantly lower (p = 0.028) after the shift. Although there was no significant increase in overall levels of parathyroid hormone (PTH) after the shift (p = 0.061), but the pre-shift and post-shift PTH level was significantly higher in IRs than in non-IRs (p = 0.003 and p = 0.027, respectively). IRs had a significantly lower initial T-CHOL (p = 0.03) and initial CRP (p = 0.012) than non-responders, which may be related to lower inflammation.ConclusionWe found the shift from DPO to CERA results in lower Al levels, a reduced inflammatory response, and an increase in RBC number and PTH level in uremic patients on hemodialysis.     

Symptoms of depression and PTSD are associated with elevated alcohol demand

BackgroundBehavioral economic demand curves measure individual differences in motivation for alcohol and have been associated with problematic patterns of alcohol use, but little is known about the variables that may contribute to elevated demand. Negative visceral states have been theorized to increase demand for alcohol and to contribute to excessive drinking patterns, but little empirical research has evaluated this possibility. The present study tested the hypothesis that symptoms of depression and PTSD would be uniquely associated with elevated alcohol demand even after taking into account differences in typical drinking levels.MethodAn Alcohol Purchase Task (APT) was used to generate a demand curve measure of alcohol reinforcement in a sample of 133 college students (50.4% male, 64.4% Caucasian, 29.5% African-American) who reported at least one heavy drinking episode (5/4 or more drinks in one occasion for a man/woman) in the past month. Participants also completed standard measures of alcohol consumption and symptoms of depression and PTSD.ResultsRegression analyses indicated that symptoms of depression were associated with higher demand intensity (alcohol consumption when price = 0; ΔR² = .05, p = .002) and lower elasticity (ΔR² = .04, p = .03), and that PTSD symptoms were associated with all five demand curve metrics (ΔR² = .04–.07, ps < .05).ConclusionsThese findings provide support for behavioral economic models of addiction that highlight the role of aversive visceral states in increasing the reward value of alcohol and provide an additional theoretical model to explain the association between negative affect and problematic drinking patterns.     

Effects of glyphosate exposure on sperm concentration in rodents: A systematic review and meta-analysis

BackgroundCorrelation between exposure to glyphosate and sperm concentrations is important in reproductive toxicity risk assessment for male reproductive functions. Many studies have focused on reproductive toxicity on glyphosate, however, results are still controversial. We conducted a systematic review of epidemiological studies on the association between glyphosate exposure and sperm concentrations of rodents. The aim of this study is to explore the potential adverse effects of glyphosate on reproductive function of male rodents.MethodsSystematic and comprehensive literature search was performed in MEDLINE, TOXLINE, Embase, WANFANG and CNKI databases with different combinations of glyphosate exposure and sperm concentration. 8 studies were eventually identified and random-effect model was conducted. Heterogeneity among study results was calculated via chi-square tests. Ten independent experimental datasets from these eight studies were acquired to synthesize the random-effect model.ResultsA decrease in sperm concentrations was found with mean difference of sperm concentrations(MDsperm) = −2.774 × 10⁶/sperm/g/testis(95%CI = −0.969 to −4.579) in random-effect model after glyphosate exposure. There was also a significant decrease after fitting the random-effect model: MDsperm = −1.632 × 10⁶/sperm/g/testis (95%CI = −0.662 to −2.601).ConclusionsThe results of meta-analysis support the hypothesis that glyphosate exposure decreased sperm concentration in rodents. Therefore, we conclude that glyphosate is toxic to male rodent’s reproductive system.     

In vivo and in vitro evaluation of the estrogenic properties of the 17β-(butylamino)-1,3,5(10)-estratrien-3-ol (buame) related to 17β-estradiol

BackgroundBuame [17β-(butylamino)-1,3,5(10)-estratrien-3-ol] possesses anticoagulant and antiplatelet activities that are potentially antithrombotic. Since its estrogenicity is unknown, it was evaluated by established methods.MethodsBuame (10, 100, 500, and 1,000 μg/kg), 17β-estradiol (E₂) (100 μg/kg), or propylene glycol (10 ml/kg) were subcutaneously (sc) administered for three days to immature Wistar female rats (21 days old). The relative uterotrophic effect to E₂ was 78 (E₂ = 100) with a relative uterotrophic potency of 1.48 (E₂ = 100). Adult ovariectomized Wistar rats received an sc injection at 8:00 h (reversed cycle) of: 7.5 μg of E₂ (≈ 30 μg/kg), buame (≈ 750, 1,500, 3,000 μg/kg), or corn oil (≈ 1.2 ml/kg). After 24 h, progesterone (4–5 mg/kg) was administered. Sexual receptivity was assessed 5 to 7 h later, and the lordosis quotient (LQ; number lordosis/number mounts × 100) was evaluated.ResultsBuame induced lordosis (LQmax 85 ± 9; ED50 952 ± 19 μg/kg) and E₂ LQmax 56 ± 8; ED50 10 ± 2 μg/kg; the relative LQpotency was 0.51 (E₂ = 100). Buame competed with [³H]E₂ for the estrogen receptor (Buame RBA = 0.15 and Ki = 5.9 × 10^?7 M; E₂ RBA = 100; Ki = 6.6 × 10^?9 M). Buame increased MCF-7 cells proliferation, from 10^?11 to 10^?9 M, its proliferative effect was 1.73–1.79 (E₂ = 3.0–3.9); its relative proliferative effect to E₂ was 33–40% (E₂ = 100%) and relative potency 10.4–10.7 (E₂ = 100). Tamoxifen and fulvestrant (ICI 182,780) inhibited buame's proliferation indicating mediation through estrogen receptors in this response.ConclusionBuame is therefore an estrogen partial agonist with a weak estrogenic activity.     

Characterization of fasted human gastric fluid for relevant rheological parameters and gastric lipase activities

PurposeTo characterize human gastric fluid with regard to rheological properties and gastric lipase activity. In addition, traditional physicochemical properties were determined.MethodsFasted HGA were collected from 19 healthy volunteers during a gastroscopic examination. Rheological characterization of the aspirates was conducted on a TA AR-G2 rheometer, using cone and plate geometry. Lipase activity was measured by continuous titration of released free fatty acid from tributyrate. Further, pH, osmolality, buffer capacity, and surface tension were measured and the total protein content and bile salt level were determined using assay kits.ResultsRheological examination of HGA showed non-Newtonian shear-thinning behavior with predominant elastic behavior in the linear range. The apparent viscosity was measured to be in the range of 1.7–9.3 mPa s at a shear rate of 50 s⁻¹. The FaSSGF and HCl pH 1.2 have no shear-thinning properties and showed lower viscosity (1.1 mPa s at 50 s⁻¹). The observed viscosity of the HGA will decrease the intrinsic dissolution rate of drugs. The activity of the gastric lipase was 7.4 ± 4.0 U/mL (N = 6, n = 3) and 99.0 ± 45.3 U/mL (N = 19, n = 3) at pH 2.8 and 5.4, respectively. pH, surface tension, buffer capacity, bile salt concentration, and osmolality were measured and compared with literature data.ConclusionThe rheological behavior and the mean apparent viscosity of HGA are significantly different from that of water and should therefore be considered important during development of gastric simulated media. Further, the activity of the HGL is active even under fasted gastric conditions and might contribute to the digestion and emulsification of lipid-based drug delivery systems in the entire gastrointestinal tract. HGL should therefore be considered in gastric evaluation of lipid-based drug delivery systems.     

Systematic investigation on the turning point of over-inflammation to immunosuppression in CLP mice model and their characteristics

Immunosuppression is involved in refractory innate and adaptive immune responses and is considered to be the predominant driving force for mortality in sepsis. The cecal ligation and puncture (CLP) model is regarded as a golden standard model for sepsis study, but the turning point of over-inflammation to immunosuppression was reported differently. Herein, systematic investigation on the turning point of over-inflammation to immunosuppression in CLP mice model was carried out. The results showed only the mortality of mice challenged with of Pseudomonas aeruginosa on Day 1 not other days after the surgery was higher than that of other mice with Sham surgery, suggesting Day 1 after the CLP surgery might be the turning point. There was very low mortality even without death in Sham mice but the mortality was 80% after mice were challenged with 2.5 × 10⁷, 5.0 × 10⁶ and 1.0 × 10⁶ CFU/10 g of Pseudomonas aeruginosa, further demonstrating Day 1 after the CLP surgery was the turning point. And, CLP mice presented low levels of pro-inflammatory and anti-inflammatory cytokines, and high bacterial loads on Day 1. Additionally, the amounts and proportion of blood cells and monocytes significantly changed, too. In conclusion, Day 1 after the CLP surgery was the turning point of over-inflammation to immunosuppression, and low levels of cytokines and high bacterial loads were the characteristics of this model on Day 1, which is significant for pharmacological investigation on sepsis.     

Influence of MRP1 G1666A and GSTP1 Ile105Val genetic variants on the urinary and blood arsenic levels of Turkish smelter workers

To understand the cellular mechanisms responsible for arsenic metabolism and transport pathways plays a fundamental role in order to prevent the arsenic-induced toxicity. The effect of MRP1 G1666A and GSTP1 Ile105Val polymorphisms on blood and urinary arsenic levels were determined in 95 Turkish smelter workers. Blood and urinary arsenic concentrations were measured by GF-AAS with Zeeman correction and gene polymorphisms were investigated by PCR-RFLP method. The mean blood and urinary arsenic levels were 21.60 ± 12.28 μg/L and 5.58 ± 4.37 μg/L, respectively. A significant association between MRP1 1666A allele and urinary arsenic levels was found (p = 0.001). GSTP1 Ile105Val polymorphism was detected not to be associated with either blood or urinary arsenic levels (p = 0.384, p = 0.440, respectively). Significant association was also detected between MRP1A^-/GSTP1Val⁻ genotypes and urinary arsenic levels (p = 0.001). This study suggested that MRP1 G1666A alone and, also, combined with GSTP1 Ile105Val were associated with inter-individual variations in urinary arsenic levels, but not with blood arsenic levels.     

Effect of the nutritional supplement ALAnerv® on the serum PON1 activity in post-acute stroke patients

BackgroundParaoxonase-1 (PON1) is one of the HDL-associated proteins which contributes to the antioxidant properties of these lipoproteins. The aim of this pilot study was to evaluate the effect of the nutritional supplement ALAnerv^® on serum PON1 activity in post-acute stroke patients undergoing rehabilitation.MethodsWe enrolled 28 post-acute stroke patients and randomly divided them into (–) ALAor (+) ALAstudy groups. All the patients underwent the same rehabilitation program and received comparable standard medications. Moreover, (+) ALA patients received ALAnerv^® for two weeks (2 pills/day). The serum PON1 activity was assessed on blood samples taken at the admission and at the discharge moments, respectively. We used paraoxon (paraoxonase activity, PONA), phenyl acetate (arylesterase activity, ARYLA) and dihydrocoumarin (lactonase activity, LACTA) as substrates, the latter activity being regarded as physiologically relevant. A control group of 14 apparently healthy subjects was also created.ResultsIn the (+) ALAgroup, LACTAsignificantly increased during the study period (17.6 ± 3.2 vs. 27.6 ± 3.5, p = 0.002). Moreover, the percentage of LACTAvariation between (–) ALAand (+) ALAgroups during the study was also statistically different (–11.7 ± 6.9% vs. +95.1 ± 29.7%, p < 0.0001).ConclusionsThese preliminary results suggest that ALAnerv^® could contribute to the improvement of the physiologically relevant LACTAof PON1 in post-acute stroke patients, enabling this enzyme to contribute to the redox correction. Also, this study raises the question about the effect of a longer treatment period over the other enzymatic activities of serum PON1.     

Anti-allergic activity of emodin on IgE-mediated activation in RBL-2H3 cells

BackgroundEmodin (1,3,8-trihydroxy-6-methylanthraquinone) is a Chinese herbal anthraquinone derivative from the rhizome of rhubarb (Rheum palmatum L.) that exhibits numerous biological activities, such as antitumor, antibacterial, antiinflammatory, and immunosuppressive. In the present studies, the anti-allergic activities of emodin were investigated to elucidate the underlying active mechanisms.MethodsThe inhibitory effects of emodin on the IgE-mediated allergic response in rat basophilic leukemia (RBL-2H3) cells were evaluated by measuring the release of granules and cytokines. The Ca²⁺ mobilization in RBL-2H3 cells loaded with the Ca²⁺-reactive fluorescent probe Fluo-4 AM was also measured by laser scanning confocal microscope.ResultsEmodin inhibited the release of β-hexosaminidase (β-HEX; IC₅₀ = 5.5 μM) and tumor necrosis factor (TNF)-α (IC₅₀ = 11.5 μM) from RBL-2H3 cells induced by 2,4-dinitrophenylated bovine serum albumin (DNP-BSA) and displayed stronger inhibition of β-HEX release than ketotifen fumarate salt (IC₅₀ = 63.8 μM). Emodin at a concentration of 12.5 μM also inhibited the DNP-BSA-induced influx of extracellular Ca²⁺ in RBL-2H3 cells.ConclusionsThese results suggested that emodin likely exhibits anti-allergic activities via increasing the stability of the cell membrane and inhibiting extracellular Ca²⁺ influx.     

The relationship of dysthymia,minor depression,and gender to changes in smoking for current and former smokers: Longitudinal evaluation in the U.S. population

BackgroundAlthough data clearly link major depression and smoking, little is known about the association between dysthymia and minor depression and smoking behavior. The current study examined changes in smoking over 3 years for current and former smokers with and without dysthymia and minor depression.MethodsParticipants who were current or former daily cigarette smokers at Wave 1 of the National Epidemiologic Survey on Alcohol and Related Conditions and completed the Wave 2 assessment were included in these analyses (n = 11,973; 46% female). Analyses examined the main and gender-specific effects of current dysthymia, lifetime dysthymia, and minor depression (a single diagnostic category that denoted current and/or lifetime prevalence) on continued smoking for Wave 1 current daily smokers and continued abstinence for Wave 1 former daily smokers.ResultsWave 1 current daily smokers with current dysthymia (OR = 2.13, 95% CI = 1.23, 3.70) or minor depression (OR = 1.53, 95% CI = 1.07, 2.18) were more likely than smokers without the respective diagnosis to report continued smoking at Wave 2. Wave 1 former daily smokers with current dysthymia (OR = 0.44, 95% CI = 0.20, 0.96) and lifetime dysthymia (OR = 0.37, 95% CI = 0.15, 0.91) were less likely than those without the diagnosis to remain abstinent from smoking at Wave 2. The gender-by-diagnosis interactions were not significant, suggesting that the impact of dysthymia and minor depression on smoking behavior is similar among men and women.ConclusionsCurrent dysthymia and minor depression are associated with a greater likelihood of continued smoking; current and lifetime dysthymia are associated with a decreased likelihood of continued smoking abstinence.     

Th17 response to Dermatophagoides pteronyssinus is related to late-phase airway and systemic inflammation in allergic asthma

BackgroundTh17 cells may play a role in the development of late-phase allergen-induced airway and systemic inflammation in allergic asthma, although the mechanisms involved remain to be elucidated.MethodsA total of 36 subjects were enrolled into the study: 15 allergic asthma patients with early asthmatic reaction (n = 7) or dual asthmatic reaction (n = 8) developed to inhaled D. pteronyssinus, 13 patients with allergic rhinitis, and 8 healthy subjects. Peripheral blood and induced sputum were collected 24 h before as well as 7 h and 24 h after a bronchial challenge with D. pteronyssinus. Th17 cells were analyzed by FACS; IL-17 levels were determined by ELISA.ResultsAt baseline, the percentage of peripheral blood Th17 cells and serum and sputum IL-17 levels were significantly higher in all groups of studied patients compared with those of healthy subjects. After the bronchial challenge, there was a significant increase in the percentage of peripheral blood Th17 cells and in serum and sputum IL-17 levels in rhinitis and asthma patients compared with their baseline values, particularly in allergic asthma patients with the dual asthmatic reaction. Positive correlations were found between the percentage of Th17 cells and IL-17 levels in serum (Rs = 0.649; P = 0.009) as well in sputum (Rs = 0.583; P = 0.022) in allergic asthma patients 24 h after the bronchial challenge.ConclusionsThe Th17 response is associated with the development of late-phase airway and systemic inflammation after the inhalation of D. pteronyssinus in patients with allergic asthma.