Content of the anti-DNA antibody in the serum of BXSB mouse and suppressive effect of BXSB mouse serum on lymphocyte proliferation

The content of the anti-DNA antibody in BXSB mouse serum and the effect of the sere from BXSB mice on lymphocyte proliferation of normal mouse were observed.Enzyme-linked immunosorbent assay (ELISA) and ~3H-thymidine incorporation were used.The content of anti-DNA antibody in male BXSB mouse serum increased from the age of 3 month and reached to the     

The relationship of serum homocysteine with collagen Ⅳ and serum cystatin C in patient with diabetic nephropathy

目的 探讨糖尿病肾病(DN)患者血清同型半胱氨酸(Hcy)水平与Ⅳ型胶原(ⅣC)、胱抑素C(CysC)的相关性.方法 150例2型糖尿病患者按照尿白蛋白水平分为正常蛋白尿(A)组、微量白蛋白尿(B)组及临床蛋白尿(C)组,每组50例;另选50例健康人作对照(D组).用ELISA法检测血清Hcy,放射免疫法检测血清ⅣC,颗粒增强免疫比浊法检测血清CysC的含量,分析血清Hcy水平与ⅣC、CysC水平的相关性.结果 2型糖尿病患者血清Hcy、ⅣC、CysC水平均高于D组(P＜0.01),B、C组高于A组(P＜0.01);血清Hcy水平与ⅣC和CysC水平呈正相关(n=0.58和r2 =0.68,P＜0.05).结论 血清Hcy导致ⅣC、CysC水平升高,参与DN的发生发展过程.     

Modulation of serum concentrations and hepatic metabolism of 17β-estradiol and testosterone by amitraz in rats

The present study has investigated the ability of amitraz, a widely used formamidine pesticide, to modulate serum concentrations and liver microsomal metabolism of 17β-estradiol (E2) and testosterone in rats. Amitraz was administered intraperitoneally to male rats for 4 days and to intact female rats or ovariectomized (OVX) and 0.5 mg/kg E2-supplemented female rats for 7 days. E2 and metabolites were analyzed by gas chromatography-electron capture detection and testosterone and metabolites were analyzed by high-pressure liquid chromatography. In OVX and E2-supplemented females, 50 mg/kg amitraz caused an 85% decrease of serum E2 concentration and a marked increase of 2-OH-E2 concentration. Amitraz at 25 and 50 mg/kg produced 9.0-fold or greater increases of serum testosterone and 2β-OH-testosterone levels in males. Amitraz at 25 mg/kg resulted in no or minimal increases of liver microsomal formation of E2 or testosterone metabolites. Amitraz at 50 mg/kg produced 1.4- to 3.6-fold increases of 2-OH-E2; estrone; 2β-, 6β-, and 16α-OH-testosterone; and androstenedione formation in males and intact females. Amitraz at 50 mg/kg preferentially increased intact female 16β-OH-testosterone production by 8.6-fold. In OVX females, E2 supplement alone or cotreatment with E2 and 50 mg/kg amitraz produced 1.3- to several-fold increases of 2- and 4-OH-E2 formation and 2β- and 16α-OH-testosterone production. The cotreatment increased 6β- and 16β-OH-testosterone formation by 1.8- and 1.6-fold, respectively. The present findings show that amitraz induces hepatic E2 and testosterone metabolism in male and female rats, decreases serum E2 concentration in OVX and E2-supplemented females, but increases serum testosterone in males.     

Absence of a relationship between sympathetic neuronal activity and turnover of serum dopamine-β-hydroxylase

Summary The effects of pharmacological alteration of adrenergic transmission on the rate of entrance of dopamine--hydroxylase (DBH) into the circulation were assessed in rats by an immunological method in which the kinetics of recovery of serum DBH activity were measured after depletion of the enzyme by treatment with anti-rat DBH antiserum. Neither -receptor blockade with phenoxybenzamine nor ganglionic blockade with clorisondamine altered the rate by which DBH enters the bloodstream although both treatments markedly altered serum catecholamine levels. Prolonged treatment of newborn rats with guanethidine produced a severe peripheral sympathectomy but only a moderate decrease (30%) in serum DBH levels. In the sympathectomized rats, the rate of entrance of DBH into the circulation was significantly reduced whereas the half-life and rate of degradation of the enzyme was unchanged. These results indicate that the major portion of serum DBH does not enter the circulation by means of exocytotic release of the soluble enzyme.     

Binding ofβ-adrenoceptor blocking drugs to human serum albumin,to α1-acid glycoprotein and to human serum

Summary The binding of 8 -adrenergic blocking drugs to human serum albumin, to ₁-acid glycoprotein and to serum from normal volunteers and from patients with rheumatoid arthritis was studied. Protein binding was determined in vitro using equilibrium dialysis of labelled drug at 25° C. Oxprenolol and propranolol were highly bound to serum, alprenolol, pindolol and timolol to a lesser degree, and atenolol, metoprolol and sotalol were negligibly bound. For the five compounds which were appreciably bound, the mean binding was significantly higher in serum from patients with rheumatoid arthritis than in serum from normal volunteers. For those drugs, binding to ₁-acid glycoprotein was higher than to human serum albumin, and binding to a mixture of both proteins approached that to serum from healthy volunteers. For each of these drugs there was a strong correlation between the serum ₁-glycoprotein concentration and the percentage binding.     

Binding of 16α-gitoxin and its 16-acetate to human serum albumin

Summary The binding of semi-synthetic (16-gitoxin and its 16-acetate) and naturally occurring cardioactive glycosides (digitoxin, ouabain) to human serum albumin was studied using equilibrium dialysis, ultracentrifugation and gel filtration. 16-gitoxin was 75% bound and its 16-acetate 95%. The percentage binding of digitoxin and ouabain was in good agreement with values reported in literature.     

Effects of OATP -C gene polymorphisms and low-dose rifampicin on serum bilirubin level

AIM OATP-C is a liver-specific organic anion uptake transporter and shows high affinity for bilirubin uptaking. Rifampicin has been identified as a potent inhibitor of OATP-C both in vitro and in vivo. This study was set to determine the allele frequencies of OATP-C*1a‘, OATP-C*1b, and OATP-C* 15 in Chinese population, and secondly, to quantitate the contribution of the OATP-C gene polymorphisms and low-dose rifampicin adminstration to the serum bilirubin level in vivo. METHODS Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and a novel tetreprimers method were used to identify OATP-C*1a, OATP-C*1b, and OATP-C*15 genotypes.     

Comparative examination of adsorption of serum proteins on HSA- and PLGA-based nanoparticles using SDS–PAGE and LC–MS

The behavior of nanosized drug carrier systems under cell culture conditions and therefore also the destiny in the body are highly influenced by the protein corona, which is formed upon entering a biological environment. Some of the adsorbed proteins, named opsonins, lead to a shortened plasma circulation half-life of the nanoparticles. Others are attributed to promote the transport of nanoparticles into other compartments of the body, just to mention two examples. Hence, detailed knowledge concerning the composition of the protein corona is of great importance. The aim of this work was to investigate the influence of the nanoparticle starting material and the surface modification on the composition of the adsorbed serum proteins in a cell culture environment. Therefore, positively charged nanoparticles based on the biodegradable polymer poly(dl-lactide-co-glycolide) (PLGA) stabilized with didodecyldimethylammonium bromide (DMAB) and negatively charged nanoparticles based on human serum albumin (HSA) were prepared and modified with hydrophilic polymers. By incubating the nanoparticles with fetal bovine serum (FBS) the adsorption of serum proteins on the colloidal system was investigated. Using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE) a semi-quantitative analysis of the protein corona was performed and after enzymatic in-solution-digestion the adsorbed proteins were identified using high resolution LC–MS. Our study accentuates the influence of the core material, surface charge, and surface modification on the amount and nature of the adsorbed proteins. The combination of SDS–PAGE and LC–MS turns out to be a simple and reliable method to investigate the protein corona of nanoparticles.     

High doses of bifendate elevate serum and hepatic triglyceride levels in rabbits and mice： animal models of acute hypertriglyceridemia

AIM: To investigate the effects of bifendate on serum and hepatic lipids level in rabbits and mice. METHODS: Animals were administered bifendate [powdered pill suspended in 0.5% sodium carboxymethylcellulose (CMC)] at increasing doses (0.25-1 g/kg, ig). Blood lipid and apolipoprotein levels were measured using commercially available assay kits. RESULTS: The treatment of rabbits with a single dose of bifendate (0.3 g/kg) caused a time-dependent and biphasic change in serum triglyceride (TG) levels, with the value reaching a maximum (3-fold increase compared to the baseline value) between 24 and 36 h post-dosing. When mice were orally treated with bifendate (0.25-1 g/kg), serum TG levels increased by 39%-76% and 14%-39% at 24 and 48 h post-dosing, respectively. When given at daily doses of 0.25 and 1 g/kg for 4 d, bifendate increased serum TG levels (56%-79%), with concomitant elevations in apolipoprotein A-I and apolipoprotein B levels at 24 h after the last dosing. TG levels were also increased (11%-43%) in liver samples of mice receiving single or multiple doses of bifendate. However, bifendate treatment caused slight reductions in serum and hepatic total cholesterol levels (9%-13%). The hypertriglyceridemia induced by bifendate was ameliorated by fenofibrate but not inositol nicotinate treatment in mice. CONCLUSION: The findings suggest that bifendate treatment at high oral doses can cause an acute elevation in serum and hepatic TG levels.     

Effective time of immune suppressive protein of stress in serum

Immune suppressive protein of stress was generated in spleen and lymph node and released into serum in mice and rots under stress,which had strong suppressive effects on T- and B-lymphocyte proliferation.The present study showed that the serum from stressed mice presented a significant decrease in     

Kinetic profile of N,N-dimethyltryptamine and β-carbolines in saliva and serum after oral administration of ayahuasca in a religious context

Ayahuasca is a beverage obtained from Banisteriopsis caapi plus Psychotria viridis. B. caapi contains the β-carbolines harmine, harmaline, and tetrahydroharmine that are monoamine oxidase inhibitors and P. viridis contains N,N-dimethyltryptamine (DMT) that is responsible for the visionary effects of the beverage. Ayahuasca use is becoming a global phenomenon, and the recreational use of DMT and similar alkaloids has also increased in recent years; such uncontrolled use can lead to severe intoxications. In this investigation, liquid chromatography–tandem mass spectrometry (LC–MS/MS) was used to study the kinetics of alkaloids over a 24 h period in saliva and serum of 14 volunteers who consumed ayahuasca twice a month in a religious context. We compared the area under the curve (AUC), maximum concentration (C_max), time to reach C_max (T_max), mean residence time (MRT), and half-life (t_1/2), as well as the serum/saliva ratios of these parameters. DMT and β-carboline concentrations (C_max) and AUC were higher in saliva than in serum and the MRT was 1.5–3.0 times higher in serum. A generalized estimation equations (GEEs) model suggested that serum concentrations could be predicted by saliva concentrations, despite large individual variability in the saliva and serum alkaloid concentrations. The possibility of using saliva as a biological matrix to detect DMT, β-carbolines, and their derivatives is very interesting because it allows fast noninvasive sample collection and could be useful for detecting similar alkaloids used recreationally that have considerable potential for intoxication.     

The influence of glutathione on the photoreaction of 3,4′,5-tribromosalicyl-anilide alone and in the presence of serum albumin

Glutathione is known to play a prominent detoxifying role in the organism. In this study attention has been paid to the possible occurrence of a detoxifying action of glutathione in the skin. For this reason we studied the influence of glutathione on the photoreaction of 3,4′,5-tribromosalicylanilide (Tbsa), which is notorious for causing photoallergy. It was found thatTbsa forms stable photoconjugates on irradiation with glutathione: 3-glutathyl-4′,5-dibromosalicylanilide and 5-glutathyl-4′-bromosalicylanilide. The rate of the photoreaction ofTbsa in the presence of glutathione, resulting in photostable products, is increased. Furthermore, covalent binding with serum albumin appeared to be decreased in the presence of glutathione.     

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).     

Characterization of the mangiferin–human serum albumin complex by spectroscopic and molecular modeling approaches

The interactions between mangiferin and human serum albumin (HSA) were investigated by spectroscopy and molecular modeling. The results proved the formation of complex between mangiferin and HSA. Hydrophobic interaction dominated in the association reaction. Mangiferin statically quenched the fluorescence of HSA in a concentration dependent manner positively deviating from the linear Scatchard equation. The binding of mangiferin to HSA lead to changes in the conformation of HSA according to synchronous fluorescence spectra, FT-IR, UV–vis and CD data. The presence of amino acids and metal ion affected the binding constant of mangiferin–HSA complex. Computational mapping of the possible binding sites of mangiferin revealed the molecule to be bound in the large hydrophobic cavity of subdomain IIA.     

Identification and characterisation of adducts between serum albumin and 4,4’-methylenediphenyl diisocyanate (MDI) in human plasma

Diisocyanates are potent inducers of airways disease. Methylenediphenyl diisocyanate (MDI) is a widely used diisocyanate in the chemical industry. The aim of this study was to identify major and also immunologically relevant protein conjugates of MDI in plasma. Plasma was obtained from an MDI-exposed worker. The plasma was dialysed and then fractionated using ion exchange chromatography (IEC) and gel filtration. These fractions and also aliquots of unfractioned plasma were hydrolysed, derivatised and analysed for isocyanate adduct content using gas chromatography–mass spectrometry. In addition, immunologically relevant proteins were identified through specific IgG immunoblotting using pooled sera from two exposed workers. It was shown by dialysis that 96% of the hydrolysed MDI derivatives were protein bound and that 95% of the MDI adducts co-eluted with serum albumin in plasma using IEC. All MDI–protein adducts co-eluted with serum albumin using gel filtration. IgG immunoblotting showed a major 66 kDa protein and also some intermolecular reactions in serum albumin. This study shows serum albumin to be the major protein in plasma that forms adducts in vivo with MDI. Thus, a quick and simple quantitative method for biological monitoring may be developed for MDI exposure. The results also showed that MDI-specific IgG antibodies preferentially bind to the serum albumin in in-vitro-synthesised MDI–plasma protein conjugates.     

Influence of continuous ambulatory peritoneal dialysis on serumα 1-acid glycoprotein concentration and drug binding

Summary The influence of continuous ambulatory peritoneal dialysis (CAPD) on the concentrations of ₁-acid glycoprotein in serum and dialysate and on the serum binding of oxprenolol, propranolol and phenytoin has been studied.Before starting CAPD treatment, the serum binding of oxprenolol and propranolol was higher and that of phenytoin lower than in healthy volunteers, and the serum ₁-AGP concentration was higher. During the first days to weeks after starting CAPD, the serum ₁-AGP concentration rose with a concomitant increase in the binding of oxprenolol and propranolol. Subsequently, the ₁-AGP level and the binding of oxprenolol and propranolol decreased to the values found before starting CAPD. The binding of phenytoin showed little change. The concentration of ₁-AGP in dialysate was 2 to 5% of that in serum.     

Pharmacokinetic properties of γ-hydroxybutyrate (GHB) in whole blood, serum, and urine

Over the last 10-15 years, γ-hydroxybutyrate (GHB) and γ-butyrolactone have become increasingly popular "club drugs", but they have also gained attention as potential agents of drug-facilitated sexual assault (DFSA). Several studies have attempted to characterize GHB's pharmacokinetic properties in humans, and the aim of this paper is to build on this research with an emphasis on DFSA cases. A 25 mg/kg dose of GHB was given to 12 GHB-na?ve volunteers (6 men and 6 women). Urine and blood samples (serum and whole blood) were collected and analyzed by gas chromatography-mass spectrometry following liquid-liquid extraction. The urinary T(max) was 1 h in 11 volunteers with a mean C(max) of 67.6 mg/L (32.6-161.3 mg/L). Urinary concentrations rapidly decreased to < 10 mg/L (interpretive limit) for 11 volunteers after just 4 h. Data derived from whole blood (mean C(max) = 48.0 mg/L, T(max) = 24.6 min) closely matched that from serum (mean C(max) = 59.4 mg/L, T(max) = 23.3 min), suggesting GHB is distributed into erythrocytes. All 12 volunteers had GHB concentrations of less than 5 mg/L in both whole blood and serum after 3 h. Results verify the rapid elimination of GHB and the limited retrospective power of a concentration-based approach to prove GHB administration in blood and urine and confirm that, in DFSA cases, samples should be collected as soon as possible.     

Protective effect of vitamin E against carbendazim-induced testicular toxicity–histopathological evidences and reduced residue levels in testis and serum

The fungicide Carbendazim Methyl-2-benzimidazole carbamate (MBC) is known to produce male reproductive toxicity. The present study has been undertaken to investigate the impact of vitamin E, an antioxidant against the testicular toxicity induced by MBC. HPLC analysis showed that the amount of MBC in testis and serum was 57.40 ± 3.38 nmol/g and 14.10 ± 0.84 nmol/ml, respectively, in rats treated with carbendazim + vitamin-E, which were significantly lower than that of rats treated with carbendazim alone (240 ± 15.60 nmol/g and 318.70 ± 22.52 nmol/ml, respectively). MBC treatment significantly decreased the testicular weight while co-administration of vitamin-E registered normal testicular weight. Histomorphometric analysis revealed a significant decrease (P < 0.05) in the diameter of the seminiferous tubules and lumen in MBC-treated rats compared to control whereas they remained normal in vitamin E + MBC-treated rats. Leydig cells appeared dispersed and hypertrophic after MBC treatment. Various histopathological changes were observed in testis of rats treated with MBC whereas these changes were absent in vitamin-E + MBC-treated rat testis. In conclusion protection against MBC-induced toxicity was observed with co-administration of vitamin E with MBC.     

Influence of polymorphisms and TNF and IL1β serum concentration on the infliximab response in Crohn’s disease and ulcerative colitis

Aim

Inflammatory bowel diseases (IBD), such as Crohn’s disease (CD) and ulcerative colitis (UC), are partially attributable to an increased secretion of proinflamatory cytokines, such as tumour necrosis factor (TNF) and interleukin-1β (IL1β), which play essential roles in the disease pathogenesis and are target molecules for specific therapy. Given the inter-individual variability in the response to the anti-TNF monoclonal antibody infliximab, the aim of our study was to explore the predictive value of TNF and/or IL1β as surrogate markers of infliximab response.

Methods

Serial serum concentrations of TNF and IL1β and TNF promoter region and IL1B polymorphisms were determined in 47 patients (29 CD and 18 UC) receiving infliximab and correlated with treatment response.

Results

Baseline serum concentrations of TNF and IL1β were higher in UC patients than in CD patients (p?=?0.0097 and?0.0024, respectively). CD patients showing <0.64 pg/ml IL1β at baseline were more frequently responders than non-responders (p?=?0.036), and the C allele of the IL1B polymorphism was associated with higher IL1β serum concentrations (p?=?0.026) and with poorer clinical remission after 14 weeks of infliximab treatment. No significant association was found between serum TNF concentration or TNF polymorphism and patient response to infliximab.

Conclusion

This is the first study evaluating the pharmacogenetic role of the rs1143634 polymorphism of IL1B and TNF polymorphisms in infliximab-treated IBD patients. We found an association between the rs1143634 C allele and higher serum IL1β concentrations and a lower response to infliximab treatment in CD patients that warrants the interest of future studies in larger and independent series.     

Validation of a liquid chromatography–tandem mass spectrometric assay for the quantitative determination of hydrastine and berberine in human serum

A high throughput liquid chromatography/tandem mass spectrometry (LC–MS/MS) method for the simultaneous determination of berberine and hydrastine in human serum, after oral administration of goldenseal (Hydrastis canadensis L.), was developed using simple acetonitrile treatment of serum samples. Noscapine served as the internal standard. Lower limit of quantification for both analytes was 0.1 ng mL⁻¹ using positive ion electrospray tandem mass spectrometry (MS/MS). The intra-day (n = 5) accuracy and precision of the method for hydrastine was 82 ± 8.8%, 97.9 ± 2.4% and 96.2 ± 3.3%, respectively. The inter-day (n = 4) accuracy and precision for hydrastine was 90.0 ± 15.17%, 99.9 ± 7.1% and 98 ± 6.54%, respectively. For berberine quantitation intra-day accuracy and precision was 96.0 ± 8.4%, 92.5 ± 4.7% and 94.4 ± 3.7%, respectively. The respective values for inter-day quantitation were 91.0 ± 8.4%, 94.3 ± 4.7% and 94.4 ± 3.7%. The analytical recovery for hydrastine was 82.4–96.2% and for berberine it was 94.4–96.0%. The analytes and noscapine were stable for 24 h at room temperature (CV 5–10%). Matrix ion effects were studied by post-column infusion of hydrastine and berberine, calculation of calibration curve slope precision was obtained using serum from five different subjects, and by comparison of the response of methanol standards and extracted serum samples. The method was further validated by determination of serum pharmacokinetics of hydrastine and berberine after administration of a single oral dose of goldenseal extract containing 77 mg of hydrastine and 132 mg of berberine.