Formulation and characterization of zein/chitosan complex particles stabilized Pickering emulsion with the encapsulation and delivery of vitamin D3.

BACKGROUND: Pickering emulsions (PEs) which are stabilized by solid particles instead of surfactants have recently attracted tremendous attentions due to their non-toxic and long-term stable nature. In the current study, we fabricated and characterized zein (ZN)/chitosan (CS) complex particles (ZNCSPs) stabilized PE for the encapsulation and delivery of vitamin D3 . RESULTS: The ZNCSPs were synthesized with different ratios, i.e. 1:1, 1:1.5 and 1:2 to investigate the optimum ratio. Transmission electron microscopy observations showed the spherical nature with smooth surface of the obtained particles in the case of ZNCS ratio 1:1.5 and 1:2. Furthermore, ζ-potential values for the these particles were 32.53 ± 1.3 and 52.86 ± 0.68 mV respectively, indicating particles with (1:2) being more stable than 1:1.5. Thereafter, using these particles, the PEs were successfully formulated with different oil (medium chain triglyceride) fractions (330, 500 and 660 g kg-1 ). The emulsions were evaluated for stability during storage and against different environmental factors including pH, temperature and ionic strength on the creaming indices (CIs) of these emulsions. The results demonstrated that the PEs with oil fractions 330 and 500 g kg-1 exhibited significant stability during storage, particularly the ones with 500 g kg-1 oil fractions which were stable against all the tested parameters. Finally, the prepared PEs were evaluated as efficient delivery system by encapsulating and delivering vitamin D3 . In vitro drug release profile confirmed sustained and controlled release of the encapsulated vitamin D3 . CONCLUSION: Overall, our findings suggest that ZNCSPs can be promising stabilizers for stable PEs that can be used as potential delivery systems in food, cosmetic and pharmaceutical industries. © 2021 Society of Chemical Industry.

Does dihydromyricetin impact on alcohol metabolism.

Dihydromyricetin (DHM) is a natural flavonoid showing several health promoting effects such as protective activity during severe alcohol intoxication. The mechanism underlying the effects of DHM on alcohol metabolism is virtually unknown. The present paper is focused on clarifying the role of DHM in the liver alcohol elimination at its molecular level. First, impact of DHM on alcohol dehydrogenase (ADH) activity in vitro and the enzyme induction in vivo was examined. Neither the ADH activity nor the enzyme expression were influenced by DHM. Next, the effect of DHM during alcohol intoxication were studied on primary hepatocytes isolated from EtOH-premedicated and untreated rats. The viability of cells exposed to alcohol, estimated based on the released enzymes, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), was slightly affected by DHM. Although the expected hepatoprotective effect of DHM was not fully achieved, DHM (in a concentration manner) proved to reduce the level of ROS/RNS in hepatocytes. However, no change in the rate of alcohol metabolism in vivo was found when rats were administered with a single or repeated dose of ethanol supplemented with DHM. In conclusion, the proposed positive effect of DHM during alcohol intoxication has not been proven. Moreover, there is no effect of DHM on the alcohol metabolism. The "hoped-for" DHM hepatoprotective activity can be attributed to the reduction of ROS/RNS levels in cells.

Differences between lipid content and composition of different parts of fillets from crossbred farmed carp (Cyprinus carpio).

Common carp (Cyprinus carpio) is, globally, one of the most important farmed fish species. We have analyzed carp from a natural pond-production system in South Bohemia. Ongrowing fish had access to an abundant plankton population which was supplemented with cereals. Fatty acid composition (FA) was investigated in three parts of fillets from four crosses of carp. The FA composition of the leanest part, the dorsal white muscle was similar to that of marine-farmed species; it contained a large proportion of n-3 highly unsaturated FA, the n-3/n-6 ratio was 1.1, and the proportion of phospholipids was high. The abdominal wall is rich in monounsaturated FA, the n-3/n-6 ratio is lower, 0.5, and it is more affected by the cereal feed. We concluded that the lipid composition of all these carp tissues can be improved in terms of healthy FA profile.

Intake of carp meat from two aquaculture production systems aimed at secondary prevention of ischemic heart disease - a follow-up study.

Our previous study showed that a diet enriched with 400 g of carp per week improved plasma lipids in subjects after aortocoronary bypass (CABG). The aim of the present study is to determine whether the different carp farming systems have an impact on the effects of carp meat in secondary cardiovascular prevention. We examined 3 groups of patients after CABG over a 4-week period of spa treatment (108 persons, 73 males, 35 females, age over 60 years). We found no differences in baseline values of blood pressure or plasma lipids. The patients were given a standard spa diet (controls; N=36) or a diet enriched of 400 g of carp meat per week, enriched omega 3 (N=37) or cereal carp (N=35). Plasma lipid parameters were examined at start and after 4 weeks in a routine laboratory setting. Group consuming omega-3 carp showed the largest decline in total cholesterol, LDL cholesterol, triglycerides and an increase in HDL cholesterol (all p<0.01). We found that carp meat from the two production systems showed significantly different effects on plasma lipids. Further trials should be performed to clarify the exact causes of the differences.

N-Methylcarbamoyl-lysine adduct in globin: a new metabolic product and potential biomarker of N, N-dimethylformamide in humans.

Metabolism of the solvents N,N-dimethylformamide (DMF) and N-methylformamide (MF) results in the formation of N-methylcarbamoyl adducts at the N-terminal valine and lysine in blood protein globin, of which the lysine adduct has so far only been reported in rats given high doses of both solvents [Mráz, J., Simek, P., Chvalová, D., Nohová, H., Smigolová, P., 2004. Studies on the methyl isocyanate adducts in globin. Chem. Biol. Interact. 148, 1-10]. Here we examined whether the lysine adduct is produced, and accessible to analysis, in humans occupationally or experimentally exposed to DMF. Globin from exposed subjects (n=35) and unexposed controls (n=5) was analyzed by two methods. Edman degradation was used as a sensitive reference method to measure the valine adduct by converting it to 3-methyl-5-isopropylhydantoin (MVH). The MVH levels in globin of the exposed subjects were in the range of 1-441 nmol/g, in controls <1 nmol/g. The principal method of globin analysis consisted of enzymatic hydrolysis with pronase to release free N(epsilon)-(N-methylcarbamoyl)lysine (MLU) and N-methylcarbamoylvaline (MVU), which were determined by HPLC/MS/MS, with no clean-up or preconcentration steps needed. For MLU, the parent and product ions were m/z 204-->173, and the limit of detection was approximately 5 nmol/g globin. MLU was found in most globins from the exposed subjects but not in the controls. A close correlation between the MLU and MVH levels (nmol/g) was observed: MLU=7+0.48 MVH (R(2)=0.84, n=32). In conclusion, MLU can be easily measured in globin of workers exposed to DMF. The findings also indicate a long-term persistence of MLU in the human body, and consequently, its potential as a biomarker of chronic exposure to DMF.

Bronchial arteries in experimental pulmonary infarction: angiographic and morphometric study.

OBJECTIVES: The aim was to investigate (1) whether collateral bronchopulmonary circulation developing due to chronic pulmonary embolism could prevent the evolution of pulmonary infarction after induction of pulmonary venous outflow impairment; and (2) how collateral bronchopulmonary circulation developed after acute embolisation of the lung with impaired pulmonary venous outflow. METHODS: Fifty two mongrel dogs were studied. Thirty six dogs were experimental animals and 16 were in a control group. Unilateral impairment of pulmonary venous outflow was induced by constriction of the left pulmonary veins in two groups of experimental dogs: (1) three months after and (2) one hour before bilateral embolisation of the pulmonary artery. All animals were killed 12 days after constriction. The size of the bronchial arteries was evaluated from angiograms. The diameter and the wall thickness of the arteries were measured during histology. RESULTS: In all experimental dogs, haemorrhagic infarctions developed distally to emboli in the left lung regardless of whether the bronchial arteries were dilated before induction of pulmonary venous constriction or whether collateral circulation started to develop after pulmonary venous constriction. Constriction of the pulmonary veins was an essential factor for pulmonary infarction to develop as no infarction developed in the embolised regions of the right lungs with intact pulmonary venous outflow. Pulmonary venous constriction alone did not cause dilatation or hypertrophy of the bronchial arteries. After pulmonary artery embolisation, the same enlargement and hypertrophy of the bronchial arteries occurred both in the left lung with previously impaired venous outflow and in the right lung with intact pulmonary veins. CONCLUSIONS: Expanded bronchopulmonary circulation did not prevent the development of infarction in the embolised region of the lung with impaired pulmonary venous outflow. Development of collateral bronchopulmonary circulation was not influenced by previously impaired pulmonary venous outflow.

2,4-toluenediisocyanate and hexamethylene-diisocyanate adducts with blood proteins: assessment of reactivity of amino acid residues in vitro.

Diisocyanates, reactive compounds used in plastics industry and potent occupational allergens, readily bind to proteins both in vitro and in vivo, however, the pattern of adducts with individual amino acids has not been investigated systematically. In this study, potential of the proteinogenic amino acid residues for carbamoylation with 2,4-toluenediisocyanate (2,4-TDI) and hexamethylenediisocyanate (HDI) was evaluated. The diisocyanates were incubated in an in vitro system (buffer pH 7.4/dioxane 50:50) with: (a) a series of Nalpha-benzyloxycarbonyl amino acids (Z-amino acids) and N-acetylcysteine (Ac-Cys), model compounds for non-N-terminal amino acids of the protein chain; (b) dipeptides Val-Phe and Asp-Phe, model compounds for N-termini of globin and albumin, respectively. Reactivity of the compounds tested, evaluated from their depletion during incubation with the diisocyanates (measured by HPLC), was in the order: Ac-Cys = Asp-Phe > Val-Phe = Nalpha-Z-Lys >> Nalpha-Z-His for 2,4-TDI, and Ac-Cys > Asp-Phe > Val-Phe = Nalpha-Z-Lys > Nalpha-Z-His > N-Z-Tyr for HDI, however, the adducts with Ac-Cys were unstable. Reactions of other amino acid residues (e.g. Ser, Thr, Met, Trp, Arg, Asn, Gln) with 2,4-TDI and HDI were not observed. Thus, N-terminal amino acids and Lys residues are likely to produce most abundant adducts with diisocyanates in proteins. Further, three amino compounds with increasing pKa values (Val-Phe, Val and Nalpha-Z-Lys) were incubated with 2,4-TDI and N-acetyl-S-[4-(2-amino)tolylcarbamoyl]cysteine, a 2,4-TDI-derived thiocarbamate with carbamoylating activity, in media with 10% and no dioxane, respectively. Here, reactivity of the amino compounds was decreasing in the order: Val-Phe > Val > Nalpha-Z-Lys, which reflects the mechanism of the amine-isocyanate reaction. The experiments also demonstrate the effect of a solvent (organic phase content) on the yield of the carbamoylation reactions.

Studies on the methyl isocyanate adducts with globin.

Isocyanates such as methylisocyanate (MIC), an intermediate in the synthesis of carbamate pesticides, or diisocyanates, used in the production of plastics, are highly reactive toxic compounds that spontaneously bind to biological macromolecules. In vivo formation of stable adducts with blood protein globin offers possibilities for biomonitoring of internal exposure to various reactive species. Thus, biomonitoring of the isocyanates through determination of their specific adducts with globin is a challenge. In this study, we characterized the adducts formed in human globin upon treatment with 100-fold molar excess of MIC. The globin was subject to enzymatic hydrolysis with pronase, and the hydrolysate was analysed by high performance liquid chromatography with positive atmospheric pressure chemical ionization mass spectrometric detection (HPLC/APCI-MS). The two major MIC adducts were those with N-terminal Val and side-chain of Lys, as confirmed by comparison with the synthetic standards. About 20 other adducts were observed, and several of them were tentatively identified using their MS and MS/MS spectra. Whereas detection of the adducts with Tyr and His was expected, the adducts with Trp and Phe, and a Lys adduct containing two MIC moieties, were probably analytical artifacts resulting from the transcarbamoylation during globin hydrolysis rather than products of direct carbamoylation. The other detected products were MIC-Val-His, derived from the N-terminal dipeptide of globin beta-chain, and dipeptides consisting of MIC-Lys attached to Gly, Val, Leu, Thr, and Glu. Failure to detect the corresponding non-modified dipeptides suggests that the pronase action may be hampered by the amino acid modification. MIC is known as a metabolic intermediate of the industrial solvents N,N-dimethylformamide (DMF) and N-methylformamide (MF) in humans and rats. The HPLC/APCI-MS analysis of globin from rats injected with DMF or MF, 1000 mg/kg, revealed the presence of the MIC adducts with both Val and Lys. The level of the Val adduct in globin from the DMF-dosed rats, determined using Edman degradation and GC/MS, was ca. 40 nmol/g, which is a level common in workers occupationally exposed to DMF. This suggests that also the Lys adduct in such human globin samples can be feasible to analysis and is therefore considered for further studies as a potential biomarker of exposure to DMF.

The evidence for conjugated mandelic and phenylglyoxylic acids in the urine of rats dosed with styrene.

Male Wistar rats were dosed intraperitoneally with styrene (400 mg/kg). Urine samples were collected over phosphate buffer, pH 6.5 for 24 h. Excretion of mandelic (MA) and phenylglyoxylic acid (PGA) amounted to 1.66 +/- 0.62 and 5.21 +/- 2.44% of dose, respectively, as determined by ion-pair HPLC. After acidic hydrolysis, the amount of MA and PGA found in urine increased to 2.10 +/- 0.84 and 6.81 +/- 3.20% (mean +/- S.D.; n = 7), respectively. A similar increase was observed after alkaline hydrolysis of urine samples. Differences between hydrolysed and non-hydrolysed samples were significant in the paired t-test (P < 0.05). Further, urine samples were fractionated by HPLC. Fractions were subjected to acidic hydrolysis and analysed by HPLC and GC/MS. Both MA and PGA were detected in the fraction which did not contain any of these metabolites before hydrolytic treatment. Thus, MA and PGA, which are used as biomarkers of exposure to styrene, form hydrolysable conjugates in the rat. At least a minor part of the total urinary MA and PGA is bound in these conjugates.

Oxidative phenotype polymorphism (P450 2D6) and metabolism of toluene.

Oxidative phenotype P-450 2D6 was examined using sparteine test in 3 groups of persons to determine if there is a coincidence in the defect of the oxidative biotransformation of sparteine and impaired oxidation of toluene, which could explain interindividual differences in the amounts of hippuric acid in the urine in exposed persons. The following groups of persons were examined: 30 rotogravure printers exposed to toluene vapors at concentrations of 8-307 ppm; 20 workers, 2 months after the cessation of the long-term exposure to toluene at concentrations of 104-1,170 ppm; 48 healthy volunteers with no exposure to toluene. Among the 98 persons 5 poor metabolizers (PMs) of sparteine were found, none in the group of printers exposed to toluene. In the experimental exposure chamber 5 PMs and 6 extensive metabolizers (EMs) were exposed to toluene concentration of 245 ppm for 5 hours. Hippuric acid and o-cresol in the urine, and toluene both in blood and in alveolar air were measured. However, no significant differences were found in either of these parameters between the PM and EM groups. Thus, the sparteine test does not appear to be applicable in the identification of persons with higher risk arising from toluene exposure.

Effect of ethanol on the urinary excretion of cyclohexanol and cyclohexanediols, biomarkers of the exposure to cyclohexanone, cyclohexane and cyclohexanol in humans.

OBJECTIVES: This study explored the acute effect of ethanol (EtOH) on the urinary excretion of cyclohexanol (CH-ol), 1,2- and 1,4-cyclohexanediol (CH-diol), biomarkers of exposure to important solvents, and chemical intermediates cyclohexanone (CH-one), cyclohexane (CH) and cyclohexanol. METHODS: Volunteers (5-8 in each group) were exposed for 8 hours either to CH-one, CH or CH-ol vapor at concentrations of about 200, 1000, and 200 mg/m3, respectively, with concomitant ingestion of EtOH (4 14-g doses taken during the exposure). Urine was collected for 72 hours and analyzed for CH-ol and CH-diols using a procedure involving acidic hydrolysis and gas chromatographic determination. RESULTS: The metabolic yields of CH-ol, 1,2-, and 1,4-CH-diol, respectively, in the exposures with EtOH were as follows: 11.3%, 36%, 23% after the exposure to CH-one, 3.1%, 15%, 8% after the exposure to CH, and 6.6%, 24%, 18% after the exposure to CH-ol. [The corresponding values obtained previously in matching experiments without EtOH were as follows: 1.0%, 39%, 18% (CH-one); 0.5%, 23%, 11% (CH); and 1.1%, 19%, 8% (CH-ol).] The excretion curves of the metabolites in the exposures with EtOH were not delayed when compared with the corresponding curves of a comparison group. CONCLUSIONS: The urinary excretion of CH-diols is much less sensitive to EtOH than that of CH-ol. It is recommended to employ CH-diols as useful and more reliable biomarkers of exposure to CH-one, CH and CH-ol in field examinations.

New findings in the metabolism of N,N-dimethylformamide--consequences for evaluation of occupational risk.

Using a novel gas chromatographic method, specific mercapturic acids produced in the biotransformation of several formamide analogues have been quantified. Thus, N-acetyl-S-(N-methylcarbamoyl)cysteine, derived from an important industrial solvent N,N-dimethylformamide, was found to be a minor metabolite in rodents but an important one in humans. Because manifestations of hepatotoxicity of formamide analogues were always linked with the production of mercapturic acids, the risk from exposure to DMF in humans appears to be higher than that estimated from toxicological experiments on laboratory animals.

The effect of thymic humoral factor upon regeneration of lymphatic and haemopoietic tissues of irradiated mice.

The study investigates the effect of humoral factor isolated from calf-thymus (thymosin) upon regeneration of lymphatic and haemopoietic tissues of female (CBA times C57BL/10ScSn)F1 mice irradiated with a whole-body exposure of 500R or 600R of gamma-rays. Thymosin was applied subcutaneously in amounts of 5 mg. per mouse per day. Two schemes of application were used: 1. The first dose given immediately after irradiation, followed by one injection daily for a total of either 3 or 9 days. 2. The first dose given 3 days prior to radiation exposure, followed by one injection daily for a total of 7 days. The number of endogenous spleen colonies, 59Fe splenic uptake, and dry spleen weight, all estimated on day 9 postirradiation, were significantly higher in both experimental groups treated with thymosin, when tested against untreated controls. If the start of the treatment preceded the irradiation, the ESC number increased as much as five times beyond the level found in the irradiated untreated group and the 59Fe uptake even exceeded the values of iron incorporation in unirradiated mice. Stimulation of lymphopoiesis induced by thymosin was reflected in an increase in 2-(14)C-thymidine incorporation into DNA of the spleen, thymus and lymph nodes, measured 72 hours after irradiation.

Carcinoma of the prostate. II. Serum activity of acid phosphatase, prostatic acid phosphatase, LDH and its isoenzymes.

In 25 patients with carcinoma of the prostate (CaP) T3 and in a comparative group of 18 patients with BPH the serum enzymes of AP, tartrate labile AP, LDH, and iso-LDH were investigated simultaneously in basal conditions and after standardized transrectal prostatic biopsy. AP, PAP as well as LDH were shown to be of small diagnostic aid. The reaction of serum enzyme levels following the standardized prostatic biopsy was the same in both CaP and BPH patients. In studying LDH-isoenzymes, we found that the third fraction was elevated in almost all patients. This change is apparently not of prostatic origin, and we could not attribute it to the concomitant diseases found in some patients.

Carcinoma of the prostate. I. Histochemical examination as an aid in evaluating prostate carcinoma.

In 26 patients with carcinoma of the prostate (CaP) the histochemical and histological characteristics of the tumour were compared with the clinical course in both untreated patients and in those receiving hormonal therapy. In the control group 16 patients with BPH were examined in the same way. It was found that histological types of CaP cannot be identified by grading. Higher frequency of very strong LDH activity in CaP was the only distinction against BPH.

[Fatal traffic accidents in the South Moravian Region during 1964-1974 and their social consequences]. / Smrtelné dopravní úrazy v Jihomoravském kraji v letech 1964 - 1974 a jejich sociální dsledky.

A statistical evaluation of 2,327 persons killed during or as a result of road accidents and post-mortem examined in the years 1964-1974 brought to light the fact that the actual number of those deceased was considerably higher than that quoted by official statistics. In this particular group there were 81 cases of people whose lives might have been saved provided they had been given immediate medical and technical aid. An examination of the social consequences of the above cases made it clear that approximately every 100 cases of adults killed in their productive age (16-60 years) were associated with 77 unilaterally orphaned children, or else that for every 100 persons killed that particular period of life there were about 5 completely orphaned children .--Any effort at eliminating the all too ofter unnecessary deaths of persons involved in road accidents then makes it absolutely imperative to establish an appropriate system enabling toe recruitment of immediate medical and technical aid.

[Effect of Stopangin mouth spray on blood alcohol levels measured by the Alcotest 7410 analyser made by Drager]. / Ovlivnení hodnoty hladiny alkoholu merené analyzátorem Alcotest 7410 fy Dräger sprayem Stopangin.

During a check-up of a driver by a police patrol in the Czech Republic by means of a breath analyzer of Dräger Co. an alcohol blood level of 0.50 g/kg was assessed. Later the driver reported that before the test he used at 30- and 5-minute intervals the drug Stopangin spray. In the Institute of Forensic Medicine the authors made an experiment which revealed that 22 minutes after administration of the drug the apparatus gave a negative result. Agreement with these conclusions was expressed also by a representative of Dräger Co. and the results were published as an expert opinion with a recommended procedure for the police of the Czech Republic in the South Moravian region.

[Overview of the changes in alcohol content of beers produced in the Czech Republic]. / Prehled aktuálních zmen v obsahu alkoholu u piv vyrábených v Ceské republice.

Paper concerns the alcohol contents of beer now produced. The previous and contemporary ways of beer signing as well as a complete list of beer sorts produced in the Czech Republic is presented.

[Use of heterotopic substitutes in the urinary bladder in children and adults]. / Uzití heterotopních náhrad mocového mechýre u detí a dospívajících.

Intestinal replacements of the urinary bladder are a solution of severe congenital damage of the lower urinary pathways also in young patients. The children tolerate the operation very well. Already at the age of four years they are able to cope with the catheterization of the stoma of the urinary reservoir. The continence of the lower urinary pathways contributes also considerably to improvement of the mental development of the child.

[Uretero-enteral anastomosis of dilated ureters in ileocecal replacement of the urinary bladder]. / Ureteroenterální anastomóza dilatovaných ureteru u ileocékální náhrady mocového mechýre.

In very dilated efferent urinary pathways with an impaired peristaltic activity of the ureter the authors selected for ureteroenteral anastomosis in ileocoecal replacement of the urinary bladder anastomosis with the ileum, leading into the ileocoecal reservoir via the invaginated trans-Bauhin's ileal valve. This valve was ensured against devagination by a transparietal suture after longitudinal intraluminal discision of the outer leaf the invaginate. By adjusting the discision of the outer leaf of the invaginate the authors obtained the selected pressure resistance of the valve which was desirable from the clinical aspect. This technique was described in two patients who were followed up for 6 and months and in a patient with convercion of Bricker's conduit into an ileocoecal prosthesis.