Increases in serum concentration of human heart-type fatty acid-binding protein following elective coronary intervention

Background: Heart-type fatty acid-binding protein (H-FABP) is considered a marker of myocardial necrosis but whether or not it is modified by myocardial ischemia is not clear. We sought to investigate if H-FABP serum levels increase following non-urgent coronary angioplasty.

Methods: We studied 31 patients undergoing coronary angioplasty. Peripheral venous samples were drawn immediately before angioplasty, 1?h after the first balloon inflation and 24?h after the procedure and assayed for H-FABP.

Results: Serum levels of H-FABP increased significantly at 1?h vs baseline from 2554?±?1268 to 3322?±?245?pg?ml^?1 (p?=?0.024). However, no differences were observed between 1?h and 24?h after angioplasty (3268?±?1861 vs 3322?±?2459?pg ml^?1, p?=?0.87). Moreover, no significant difference was observed when we compared 24?h after angioplasty with the baseline (3268?±?1861vs 2554?±?1268?pg ml^?1, p?=?0.112).

Conclusions: We conclude that H-FABP significantly increases after elective coronary angioplasty at 1?h compared with baseline values; whether or not this has any prognostic significance for future events, as it occurs with troponins, needs to be studied further.     

Characteristics of a nickel–albumin binding assay for assessment of myocardial ischaemia

Background: The aim of this study was to describe a method to measure ischaemia-induced alterations of the binding capacity of serum albumin to exogenous nickel.

Methods: We measured the levels of cardiac troponin I (cTnI), serum albumin, ischaemia-modified albumin (IMA) measured by a cobalt–albumin binding assay (CABA), and a nickel–albumin binding assay (NABA) in the following groups: myocardial infarction (n?=?32) and non-ischaemic chest pain (n?=?64).

Results: IMA, cTnI and NABA levels were higher in the myocardial infarction group. NABA presented a higher ability to discriminate myocardial ischaemia than CABA.

Conclusions: Patients with myocardial infarction have reduced nickel binding to human serum albumin, and NABA may have an important role as an early marker of myocardial ischaemia.     

Activity and function in lung injuries due to sulphur mustard

Aim. Pulmonary complications are known to occur in over half the patients exposed to sulphur mustard. Many studies have focused on the clinical complications, often ignoring the pathogenesis of sulphur mustard. Also, the reasons for the variable severity of lung injuries caused by sulphur mustard are unclear. Hence, the current study was performed to evaluate the correlation between superoxide dismutase (SOD) and catalase (CAT) activity and pulmonary function in patients exposed to sulphur mustard. Methods. Our study was a comparative cross-sectional survey. Two hundred and fifty incident survivors were selected from the Sardasht population who were exposed to sulphur mustard in 1987. A control group from non-exposed civilians was also selected. We used a pulmonary function test, and SOD and CAT activity was measured in these groups. Results. The mean SOD activity in the healthy control group (70.5±10.8 U ml^?1) was higher than in the moderate-to-severe group (67.0±6.1 U ml^?1) (p <0.001, one-tail ANOVA, least significant difference (LSD) post hoc). The mean activity in the mild group (72.5±6.9 U ml^?1) was no higher than in the healthy control group (70.5±10.8 U ml^?1) (p=0.095 one-tail ANOVA, LSD post hoc). The mean CAT activity in the healthy control group (4.9±1.5 U ml^?1) was lower than in the moderate-to-severe group (8.0±1.8 U ml^?1) (p <0.001, one-tail ANOVA, LSD post hoc) and in the mild group (7.5±1.5 U ml^?1) (p=0.012 one-tail ANOVA, LSD post hoc). Conclusion. According to our findings, it is reasonable to hypothesize that re-establishment of the activation–inactivation or oxidant–antioxidant balance in favour of the activation and antioxidant balances would be useful as a therapeutic strategy to suppress pathological mechanisms underlying lung injuries.     

Analysis of myosmine,cotinine and nicotine in human toenail,plasma and saliva

Myosmine is a minor tobacco alkaloid with widespread occurrence in the human diet. Myosmine is genotoxic in human cells and is readily nitrosated and peroxidated yielding reactive intermediates with carcinogenic potential. For biomonitoring of short-term and long-term exposure, analytical methods were established for determination of myosmine together with nicotine and cotinine in plasma, saliva and toenail by gas chromatography–mass spectrometry (GC/MS). Validation of the method with samples of 14 smokers and 10 non-smokers showed smoking-dependent differences of myosmine in toenails (66?±?56 vs 21?±?15?ng?g^?1, p?<0.01) as well as saliva (2.54?±?2.68 vs 0.73?±?0.65?ng ml^?1, p <0.01). However, these differences were much smaller than those with nicotine (1971?±?818 vs 132?±?82?ng g^?1, p <0.0001) and cotinine (1237?±?818 vs <35?ng?g^?1) in toenail and those of cotinine (97.43?±?84.54 vs 1.85?±?4.50?ng ml^?1, p <0.0001) in saliva. These results were confirmed in plasma samples from 84 patients undergoing gastro-oesophageal endoscopy. Differences between 25 smokers and 59 non-smokers are again much lower for myosmine (0.30?±?0.35 vs 0.16?±?0.18?ng?ml^?1, p <0.05) than for cotinine (54.67?±?29.63 vs 0.61?±?1.82?ng ml^?1, p <0.0001). In conclusion, sources other than tobacco contribute considerably to the human body burden of myosmine.     

Soluble tumour necrosis factor-α receptor type 1 as a biomarker of response to phototherapy in patients with psoriasis

Abstract

The purpose of the study was to analyze the relationship between the serum concentration of soluble tumour necrosis factor-α type 1 (sTNF-R1), the severity of plaque-type psoriasis and therapeutic response. We compared sTNF-R1 in 25 patients treated with narrowband ultraviolet B (NB-UVB) radiation and 25 patients treated with systemic photochemotherapy (psoralen plus UVA – PUVA). The pretreatment Psoriasis Area and Severity Index (PASI) score and sTNF-R1 concentration were 16.32±5.26 and 1.99±0.40 ng ml^?1, respectively, in the group treated with NB-UVB, and 17.22±3.48 and 2.07±0.31 ng ml^?1, respectively, in the group treated with PUVA. The concentration of sTNF-R1 in healthy controls was 1.49±0.34 ng ml^?1 (p<0.05 compared with patients with psoriasis). The pretreatment PASI score correlated with sTNF-R1 in both treatment groups (r=0.46 and r=0.44, p<0.05). NB-UVB and PUVA gave similar therapeutic effects (the PASI score after 20 treatments was 4.42±1.67 in the NB-UVB-treated group and 5.55±2.10 in PUVA-treated patients); however, the sTNF-R1 concentration at the same time differed significantly: 1.52±0.37 ng ml^?1 and 1.98±0.39 ng ml^?1 (p<0.001), respectively. Moreover, the decline in sTNF-R1 in both treatment groups was significant only in patients in whom the duration of skin lesions was less than 3 months. The results suggest that the value of serum sTNF-R1 concentration as a marker of response to phototherapy may depend on duration of skin lesions and the treatment method.     

Biomarker-specific differences between transpulmonary and peripheral arterial–venous blood sampling in patients with pulmonary hypertension

Abstract

Purpose: Transpulmonary biomarkers may provide insight into pulmonary hypertension (PH) pathophysiology, but require cardiac catheterization. We investigated whether the peripheral arterial–venous ratio (PR) could substitute for the transpulmonary ratio (TPR).

Materials and methods: Blood from the pulmonary artery (PA), pulmonary arterial wedge (PAW), peripheral venous, and peripheral arterial positions was analysed for ET-1, NT-pro-BNP and cAMP levels in subjects with no PH (n?=?18) and PH due to left heart disease (PH-LHD), which included combined pre- and post-capillary PH (Cpc-PH; n?=?7) and isolated post-capillary PH (Ipc-PH; n?=?9). Bland–Altman comparisons were made between peripheral venous and PA samples and between peripheral arterial and PAW samples. TPR was defined as [PAW]/[PA].

Results: For ET-1, Bland–Altman analysis indicated negative bias (?24%) in peripheral arterial compared to PAW concentration and positive bias (23%) in peripheral venous compared to PA concentration. There was <10% absolute bias for NT-pro-BNP and cAMP. For ET-1, there was no difference in PR between Cpc-PH and Ipc-PH (0.87?±?0.4 vs. 0.94?±?0.6, p?=?0.8), whereas there was a difference in TPR (2.2?±?1.1 vs. 1.1?±?0.2, p?<?0.05).

Conclusions: In PH-LHD, peripheral samples may be inadequate surrogates for transpulmonary samples, particularly when measuring mediators with prominent pulmonary secretion or clearance, such as ET-1.     

Kocuria SM1 controls vibriosis in rainbow trout (Oncorhynchus mykiss,Walbaum)

Aims: To develop probiotics for the control of vibriosis caused by Vibrio anguillarum and Vibrio ordalii in finfish. Methods and Results: Kocuria SM1, isolated from the digestive tract of rainbow trout, was administered orally to rainbow trout (Oncorhynchus mykiss) for 2 weeks at a dose equivalent to c. 10⁸ cells per g of feed and then challenged intraperitoneally with V. anguillarum and V. ordalii. Use of SM1 led to a reduction in mortalities to 15–20% compared to 74–80% mortalities in the controls. SM1 stimulated both cellular and humoral immune responses in rainbow trout, by elevation of leucocytes (5·5 ± 0·8 × 10⁶ ml^?1 from 3·7 ± 0·8 × 10⁶ ml^?1), erythrocytes (1·2 ± 0·1 × 10⁸ ml^?1 from 0·8 ± 0·1 × 10⁸ ml^?1), protein (23 ± 4·4 mg ml^?1 from 16 ± 1·3 mg ml^?1), globulin (15·7 ± 0·2 mg ml^?1 from 9·9 ± 0·1 mg ml^?1) and albumin (7·3 ± 0·2 mg ml^?1 from 6·1 ± 0·1 mg ml^?1) levels, upregulation of respiratory burst (0·05 ± 0·01 from 0·02 ± 0·01), complement (56 ± 7·2 units ml^?1 from 40 ± 8·0 units ml^?1), lysozyme (920 ± 128·8 units ml^?1 from 760 ± 115·3 units ml^?1) and bacterial killing activities. Conclusions: Kocuria SM1 successfully controlled vibriosis in rainbow trout, and the mode of action reflected stimulation of the host innate immune system. Significance and Impact of the Study: Probiotics can contribute a significant role in fish disease control strategies, and their use may replace some of the inhibitory chemicals currently used in fish farms.     

Content of cysteine, reduced and oxidized glutathione in spermatozoa of representatives of Acipenseriformes (Acipenser baerii and A. ruthenus) as well as teleosts (Perca fluviatilis and Sander lucioperca)

Glutathione belongs to a vital intra‐ and extra‐cellular protective antioxidant and is found almost exclusively in its reduced form. The ratio between its reduced and oxidized within cells is often used as a marker of cellular toxicity. The objectives of the study were to (i) determine both the reduced (GSH) and oxidized glutathione (GSSG) and cysteine (Cys) in the sperm of the Acipenser baerii and Acipenser ruthenus, as well as in Perca fluviatilis and Sander lucioperca, and (ii) to demonstrate the differences in concentration levels between representatives of acipenseriform and teleost species. High performance liquid chromatography with electrochemical detection was employed. The average content of the thiols determined in the sperm samples were as follows: Acipenser baerii (cysteine 55 ± 8 μg ml^?1; GSH 126 ± 19 μg ml^?1; GSSG 49 ± 7 μg ml^?1), Acipenser ruthenus (cysteine 62 ± 9 μg ml^?1; GSH 768 ± 115 μg ml^?1; GSSG 180 ± 16 μg ml^?1), Sander lucioperca (cysteine 251 ± 38 μg ml^?1; GSH 185 ± 28 μg ml^?1; GSSG 93 ± 14 μg ml^?1), Perca fluviatilis (cysteine 281 ± 42 μg ml^?1; GSH 496 ± 74 μg ml^?1; GSSG 138 ± 21 μg ml^?1). Based on the results obtained it can be concluded that this method is sensitive and selective for the determination of these compounds in real samples. Results revealed differences in cysteine content between species of the two systematic categories but also showed that ratios between GSH and GSSG can vary between species while potentially predict oxidative stress in fish sperm.     

Ischemia-modified albumin as a marker of vascular dysfunction and subclinical atherosclerosis in β-thalassemia major

Background: Ischemia-modified albumin (IMA) is an altered type of serum albumin that forms under conditions of oxidative stress and an independent predictor of major adverse cardiovascular events.

Objectives: To measure the levels of IMA in 45 children and adolescents with β-thalassemia major (β-TM) compared with 30 healthy controls and assess its relation to lipid peroxidation, vascular complications and subclinical atherosclerosis.

Methods: β-TM patients without symptoms of heart disease were studied focusing on transfusion history, chelation therapy, serum ferritin, malondialdehyde (MDA) and IMA levels. Echocardiography was performed and carotid intima media thickness (CIMT) was assessed.

Results: IMA and MDA levels were significantly higher in β-TM patients compared with controls (p?Conclusion: Our results highlight the role of oxidative stress in the pathophysiology of vascular complications in thalassemia. IMA could be useful for screening of β-TM patients at risk of cardiopulmonary complications and atherosclerosis because its alteration occurs in early subclinical disease.     

Forskolin-loaded human serum albumin nanoparticles and its biological importance

Abstract

In this study, forskolin-loaded human serum albumin nanoparticles (FR-HSANPs) were successfully prepared by incorporation and affinity-binding methods. FR-HSANPs were characterized by transmission electron microscope that most of them are circular in shape and size is around 340?nm. The drug loading was more than 88% and further sustained release profiles were observed as it is 77.5% in 24?h time. Additionally, the cytotoxicity results with HepG2 cells indicated that FR-HSANPs showed significantly higher cytotoxicity and lower cell viability as compared to free forskolin (FR). Furthermore, to understand the binding mechanism of human serum albumin (HSA) with forskolin resulted from fluorescence quenching as a static mechanism and the binding constant is 6.26?±?0.1?×?10⁴ M^?1, indicating a strong binding affinity. Further, association and dissociation kinetics of forskolin–HSA was calculated from surface plasmon resonance spectroscopy and the binding constant found to be K_forskolin = 3.4?±?0.24?×?10⁴ M^?1 and also fast dissociation was observed. Further, we used circular dichroism and molecular dynamics simulations to elucidate the possible structural changes including local conformational changes and rigidity of the residues of both HSA and HSA–forskolin complexes.

Communicated by Ramaswamy H. Sarma     

Decreased paraoxonase 2 enzymatic activity in monocyte/macrophages cells. A comparative in vivo and in vitro study for diabetes

Aim: To investigate peripheral blood monocytes/macrophages (Mo/M?) paraoxonase 2 (PON2) in diabetes and the factors modulating its activity.

Methods: One hundred and eighteen patients with newly diagnosed uncomplicated type 2 diabetes mellitus were compared regarding clinical, biochemical and oxidative stress parameters with 80 healthy subjects. The capacity of the peripheral blood mononuclear cells (PBMNC) to release pro-oxidants and to neutralise them was determined by measuring the respiratory burst (RB) and the intracellular antioxidant enzyme PON2. In vitro experiments were conducted on a differentiated monocytes cell line (dU937) that was exposed to serum deprivation followed by addition of isolated lipoproteins (VLDL or LDL).

Results: Paraoxonase 2 activity in Mo/M? was significantly lower in type 2 diabetes patients (0.042?±?0.044 vs 0.165?±?0.133U lactonase activity/mg protein in controls, p?1c) and insulin resistance (HOMA-IR). In multivariate regression models, 15–34% of the PON2 variance was explained by diabetes. The in vitro addition of VLDL normalised the RB of serum deprived dU937 cells, S? (to 82?±?18% of the cells incubated with serum, S+) and PON2 activity (from 0.524?±?0.061 in S???to 0.298?±?0.048?U/mg protein). In contrast, when LDL was added, the RB remained lower (61?±?12% of S+, p?=?.03) and PON2 higher (0.580?±?0.030?U/mg protein, p?=?.003).

Conclusions: The decrease in monocyte/macrophage PON2 enzymatic activity observed in type 2 diabetes cannot be totally explained by abdominal obesity and insulin resistance. The underlying molecular mechanisms need to be identified.     

The clinical efficacy of angiotensin II type1 receptor blockers on inflammatory markers in patients with hypertension: a multicenter randomized-controlled trial; MUSCAT-3 study

Purpose: The purpose of present study was to evaluate the clinical efficacy of irbesartan on the anti-inflammatory and anti-oxidative stress effect in patients with hypertension compared to other ARBs. Further, we assessed the effect of the ARBs on kidney function and urinary albumin excretion.

Methods: Eighty-five outpatients with hypertension who took an ARB except irbesartan more than 3?months were assigned into two groups, one continued the same ARB and the other switched the ARB to irbesartan for 6?months.

Results: Although blood pressures were equally controlled (continue group: 148?±?2/79?±?2?mmHg to 131?±?2/74?±?2?mmHg; switch group: 152?±?2/81?±?2?mmHg to 132?±?2/74?±?2?mmHg; p?<?0.001 each), the inflammatory markers (hsCRP, PTX3, MCP-1) and oxidative stress marker (MDA-LDL) did not change after 6?months in both groups. Urinary albumin excretion was significantly reduced only in the switch group without renal function deterioration (switch group 292.4?±?857.9?mg/gCr to 250.6?±?906.5?mg/gCr, p?=?0.012).

Conclusion: These results provide knowledge of the characteristics of irbesartan, suggesting appropriate choice of ARBs in the treatment for hypertension should be considered.     

Pudendal and median nerve sensory perception threshold: A comparison between normative studies

Background: For the evaluation of sensory innervation, normative data are necessary as a comparison.

Aims: To compare our current perception thresholds (CPTs) with normative data from other research.

Methods: Healthy volunteers were assessed for 2000, 250, and 5?Hz CPTs of the median and pudendal nerve and data were compared with other studies.

Results: Normative data in the studied group n?=?41 (male: 21; female: 20) for the median nerve, 2?kHz, 250?Hz, and 5?Hz were respectively: 241.85?±?67.72 (140–444); 106.27?±?39.12 (45–229); 82.05?±?43.40 (13–271). Pudendal nerve CPTs 250?Hz were: 126.44?±?69.46 (6–333). For men 2?kHz: 349.95?±?125.76 (100–588); 5?Hz: 132.67?±?51.81 (59–249) and women 2?kHz: 226.20?±?119.65 (64–528); 5?Hz: 92.45?±?44.66 (35–215). For the median nerve no statistical differences for gender were shown. For the pudendal nerve, only 250?Hz showed no difference for gender (t-test: 0.516). Comparison of our data with CPTs of other normative data showed no agreement for the pudendal nerve. For the median nerve only 2?kHz showed agreement in three studies and for 5?Hz with one study.

Conclusion: Comparing normative data of multiple studies shows a variety of results and poor agreement. Therefore, referring to normative data of other studies should be handled with caution.     

Degradation and Detoxification of Nicosulfuron by a Pseudomonas Strain Isolated from a Contaminated Cornfield Soil

ABSTRACT

The dissipation and detoxification of nicosulfuron (NS) by Pseudomonas aeruginosa B9 isolated from a cornfield soil was investigated. The fastest decline of NS occurred at 40 µg ml^?1 in liquid media with 0.25% glucose plus 0.05% yeast extract (DT₅₀ = 4 days) with a notable pH reduction (pH ? 5). Bioassay tests showed considerable phytotoxicity of NS for Cress (Lepidium sativum L.) with 50% shoot growth inhibition (SGI) at 40 µg ml^?1. The dissipation of NS (40 µg ml^?1) by the B9 isolate reduced the SGI significantly (SGI: up to 45 ± 3%) compared to the non-inoculated media (SGI: up to 58 ± 4%). In soils with the B9 isolate, NS dissipation, especially at 0.3 µg g^?1, was faster with a more significant SGI reduction (k = 0.08 ± 0.00 day^?1; SGI = 2 ± 1%) compared to non-inoculated samples (k = 0.03 ± 0.00 day^?1; SGI = 8 ± 1%). NS initially inhibited soil respiration, microbial biomass carbon, and dehydrogenase activity. The effect was however transient, and these parameters recovered within 10 days, especially in the presence of the isolate. Overall, this study proves Pseudomonas aeruginosa B9 as a suitable candidate for bioremediation of NS in contaminated sites.     

Effects of salicylic acid on post-ischaemic ventricular function and purine efflux in isolated mouse hearts

Abstract

Acetyl salicylic acid (aspirin) is one of the most widely used drugs in the world. Various plasma concentrations of aspirin and its predominant metabolite, salicylic acid, are required for its antiarthritic (1.5–2.5 mM), anti-inflammatory (0.5–5.0 mM) or antiplatelet (0.18–0.36 mM) actions. A recent study demonstrated the inhibitory effects of both aspirin and salicylic acid on oxidative phosphorylation and ATP synthesis in isolated rat cardiac mitochondria in a dose-dependent manner (0–10 mM concentration range). In this context, the present study was conducted to determine the effects of salicylic acid on inosine efflux (a potential biomarker of acute cardiac ischaemia) as well as cardiac contractile function in the isolated mouse heart following 20 min of zero-flow global ischaemia. Inosine efflux was found at significantly higher concentrations in ischaemic hearts perfused with Krebs buffer fortified with 1.0 mM salicylic acid compared with those without salicylic acid (12575±3319 vs. 1437±348 ng ml^?1 min^?1, mean±SEM, n=6 per group, p<0.01). These results indicate that 1.0 mM salicylic acid potentiates 8.8-fold ATP nucleotide purine catabolism into its metabolites (e.g. inosine, hypoxanthine). Salicylic acid (0.1 or 1.0 mM) did not appreciably inhibit purine nucleoside phosphorylase (the enzyme converts inosine to hypoxanthine) suggesting the augmented inosine efflux was due to the salicylic acid effect on upstream elements of cellular respiration. Whereas post-ischaemic cardiac function was further depressed by 1.0 mM salicylic acid, perfusion with 0.1 mM salicylic acid led to a remarkable functional improvement despite moderately increased inosine efflux (2.7-fold). We conclude that inosine is a sensitive biomarker for detecting cardiac ischaemia and salicylic acid-induced effects on cellular respiration. However, the inosine efflux level appears to be a poor predictor of the individual post-ischaemic cardiac functional recovery in this ex vivo model.     

Renalase in children with chronic kidney disease

Abstract

Background: Renalase is kidney-derived molecule initially considered as catecholamine-inactivating enzyme. However, recent studies suggest that renalase exerts potent cardio- and nephroprotective actions, not related to its enzymatic activity.

Purpose: To assess renalase level in children with chronic kidney disease (CKD).

Material and methods: Serum renalase, BMI, arterial stiffness, peripheral and central blood pressure, intima-media thickness (IMT), medications, and biochemical parameters were analyzed in 38 children with CKD (12.23?±?4.19?years) (stage G2-5). Control group consisted of 38 healthy children.

Results: In the study group, GFR was 25.74?±?8.94?mL/min/1.73 m²; 6 children were dialyzed; 26 had arterial hypertension. Renalase level was higher in the study group compared to control group (p?<?0.001). In CKD children renalase correlated (p?<?0.05) with BMI Z-score (r?=?–0.36), alfacalcidol dose (r?=?0.41), GFR (r?=?–0.69), hemoglobin (r?=?–0.48), total cholesterol (r?=?0.35), LDL-cholesterol (r?=?0.36), triglycerides (r?=?0.52), phosphate (r?=?0.35), calcium-phosphorus product (r?=?0.35), parathormone (r?=?0.58), and pulse wave velocity Z-score (r?=?0.42). In multivariate analysis GFR (β?=?–0.63, p?<?0.001), triglycerides (β?=?0.59, p?=?0.002), and alfacalcidol dose (β?=?–0.49, p?=?0.010) were determinants of renalase.

Conclusions: In children with CKD there is a strong correlation between renalase level and CKD stage. Furthermore, in these patients renalase does not correlate with blood pressure but may be a marker of arterial stiffness.     

Liposomal hydrogel formulation for transdermal delivery of pirfenidone

Context: Pirfenidone (PFD) is an anti-fibrotic and anti-inflammatory agent indicated for the treatment of idiopathic pulmonary fibrosis (IPF). The current oral administration of PFD has several limitations including first pass metabolism and gastrointestinal irritation.

Objective: The aim of this study is to investigate the feasibility of transdermal delivery of PFD using liposomal carrier system.

Materials and methods: PFD-loaded liposomes were prepared using soy phosphatidylcholine (SPC) and sodium cholate (SC). Encapsulation efficiency (EE) of PFD in liposomes was optimized using different preparation techniques including thin film hydration (TFH) method, direct injection method (DIM) and drug encapsulation using freeze–thaw cycles. In vitro drug release study was performed using dialysis membrane method. The skin permeation studies were performed using excised porcine ear skin model in a Franz diffusion cell apparatus.

Results and discussion: The average particle size and zeta-potential of liposomes were 191?±?4.1?nm and ?40.4?±?4.5?mV, respectively. The liposomes prepared by TFH followed by 10 freeze–thaw cycles showed the greatest EE of 22.7?±?0.63%. The optimized liposome formulation was incorporated in hydroxypropyl methyl cellulose (HPMC) hydrogel containing different permeation enhancers including oleic acid (OA), isopropyl myristate (IPM) and propylene glycol (PG). PFD-loaded liposomes incorporated in hydrogel containing OA and IPM showed the greatest flux of 10.9?±?1.04?μg/cm²/h across skin, which was 5-fold greater compared with free PFD. The cumulative amount of PFD permeated was 344?±?28.8?μg/cm² with a lag time of 2.3?±?1.3?h.

Conclusion: The hydrogel formulation containing PFD-loaded liposomes can be developed as a potential transdermal delivery system.     

Preparation monoclonal β-type anti-idiotype antibody of zearalenone and development of green ELISA quantitative detecting technique

Abstract

Immunoassay has been widely used in the screening of mycotoxins, which may be hazardous to the operator or the environment. This study was to develop a green way to measure zearalenone (ZEN) with a monoclonal β-type anti-idiotype antibody (Ab2β) against ZEN in place of ZEN standard. Six monoclonal β-type anti-idiotype antibodies were prepared. The 50% inhibitory concentration (IC₅₀) value to ZEN of the six antibodies was between 34.45?±?1.12–182.12?±?15.40?nM. A green ELISA was then developed and validated. The quantitative conversion formula between ZEN and the monoclonal Ab2β against ZEN was y?=?0.092x^0.722, R² = 0.990. The working range was 2.63–100.64?ng ml^?1. The recovery rate in spiked feed samples was from 82.15% to 102.79%, and the within-assay and between-assay coefficient variation (CV) level were less than 10.00%. A good correlation was obtained by high-performance liquid chromatography method (HPLC) to validate the developed method.