Human metabolism and urinary excretion kinetics of di-n-butyl adipate (DnBA) after oral and dermal administration in three volunteers.

Di-n-butyl adipate (DnBA) is used as a plasticizer and in various consumer products (e.g. personal care products) replacing, in part, the endocrine disruptor di-n-butyl phthalate (DnBP). We provide quantitative in vivo data on human DnBA metabolism and excretion after oral dose (105-185 µg/kg bw) and dermal application to three volunteers each as a tool for exposure and risk assessment. Complete and consecutive urine samples were collected for two (oral) and four days (dermal), respectively, and analyzed for the metabolites mono-n-butyl adipate (MnBA), 3- and tentative 4-hydroxy-mono-n-butyl adipate (3OH-MnBA, 4OH-MnBA), and 3-carboxy-mono-n-propyl adipate (3cx-MnPrA), as well as the hydrolysis product adipic acid (AA) using stable isotope dilution quantification. Metabolites were excreted within 24 h after oral dose with one or two concentration maxima at 0.8-3.0 h (n = 3) and 4.8-6.3 h (n = 2). AA was the major but unspecific metabolite with urinary excretion fractions (FUEs) of 14-26 %. Mean FUEs (range) of 3cx-MnPrA, MnBA, 3OH-MnBA, and tentative 4OH-MnBA were low, but consistent between volunteers (0.47 % (0.35-0.63 %), 0.079 % (0.065-0.091 %), 0.012 % (0.006-0.016 %), and 0.005 % (0.002-0.009 %), respectively). MnBA and 3OH-MnBA seem to be suitable, specific exposure biomarkers for DnBA, whereas 3cx-MnPrA and 4OH-MnBA seem to originate also from other, unknown sources not related to DnBA. Compared to the oral study, metabolite excretion in the dermal study was delayed and MnBA excretion was somewhat higher compared to the oxidized metabolites. Based on urinary concentrations and the above excretion fractions, calculated uptakes in the dermal study did not exceed the adipate ester ADI of 5 mg/(kg bw*day).

Metabolism and urinary excretion kinetics of di(2-ethylhexyl) adipate (DEHA) in four human volunteers after a single oral dose.

Di(2-ethylhexyl) adipate (DEHA) is used as a substitute for the reprotoxic phthalate plasticizer di(2-ethylhexyl) phthalate (DEHP). This study reports the first quantitative data on human in vivo DEHA metabolism and urinary metabolite excretion with the aim of providing tools for DEHA exposure and risk assessments. After DEHA was administered to four healthy volunteers (107-164 µg/kg body weight (bw)), urine samples were continuously and completely collected for 48 h and analyzed for the specific oxidized monoester metabolites mono-2-ethyl-5-hydroxyhexyl adipate (5OH-MEHA), mono-2-ethyl-5-oxohexyl adipate (5oxo-MEHA), and mono-5-carboxy-2-ethylpentyl adipate (5cx-MEPA), as well as for the non-specific hydrolysis product adipic acid (AA) using stable isotope dilution analysis. AA was confirmed as a major (urinary excretion fraction (FUE): 10-40%), yet non-specific DEHA metabolite. 5cx-MEPA was the major specific DEHA metabolite with an FUE of 0.20% (range: 0.17-0.24%). FUEs for 5OH-MEHA and 5oxo-MEHA were 0.07% (0.03-0.10%) and 0.05% (0.01-0.06%), respectively. The three specific metabolites were excreted with two concentration maxima (tmax1 = 1.5-2.3 h, tmax2 = 3.8-6.4 h). Elimination half-lives (t1/2, calculated after the second tmax) for 5cx-MEPA were calculated between 2.1-3.8 h. The majority (98-100%) of metabolites was excreted within 24 h. The FUE of 5cx-MEPA was applied to demonstrate its applicability for calculating daily intakes based on urinary metabolite levels from three pilot populations. Daily intakes were generally far below the tolerable daily intake (TDI) for DEHA (300 µg/kg bw/day). The highest daily intake (114 µg/kg bw/day) was calculated in individuals after consuming food that had been wrapped in DEHA containing cling film.

Systemic toxicity of di (2-ethylhexyl) adipate (DEHA) in rats following 28-day intravenous exposure.

Di (2-ethylhexyl) adipate (DEHA) is a potential plasticizer alternative for di-2-ethylhexyl phthalate (DEHP). Toxicity of DEHA has been studied mostly via oral exposure but not assessed after repeated intravenous exposure. The present study shows the toxicity effects after intravenous administration for 28 consecutive days and the reversibility of the effects following a 14-day recovery period. The study was conducted under GLP conditions. Four groups of rats (15/sex/group) each received either vehicle or DEHA in vehicle (100, 200, or 450 mg/kg/day). Criteria for evaluation included clinical observations, body weight, food consumption, clinical pathology (hematology, serum chemistry, coagulation, urinalyses), gross (necropsy) evaluation, organ weight and histopathological evaluation. There were no DEHA-related changes in all the endpoints evaluated at 100 or 200 mg/kg/day. There were no test article-related changes in clinical pathology or gross necropsy observation at 450 mg/kg/day. At the high-dose, DEHA-related findings included clinical observations, decreased body weight gain and food consumption, increased liver weight in females associated with minimal hepatocellular hypertrophy, and decreased thymus weight in males and females without histopathology findings. All these findings were completely reversible within a 14-day recovery period. Therefore, the 200 mg/kg/day dose is considered to be the No-Observed-Effect Level (NOEL).

Heparin appended ADH-anionic polysaccharide nanoparticles for site-specific delivery of usnic acid.

The intention of present research work is to formulate usnic acid (UA) loaded heparin modified gellan gum (HAG) nanoparticles (NPs). HAG copolymer based conjugation was synthesized and characterized by 1H NMR and FT-IR spectroscopy. Plain and UA loaded HAG NPs were prepared via nanoprecipitation technique. NPs were typified and further characterized for particle size, polydispersity index, entrapment efficiency, zeta potential, atomic force microscopy, differential scanning calorimetry, X-ray diffraction analysis, and in-vitro release. In-vitro tube formation assay, tumorsphere assay, autophagy assay, DNA cleavage assay, internalization by confocal and FACS based internalization analysis, caspase assay and cell cycle assay were performed for biological activity. Obtained experimental results explored that HAG NPs displayed a sustained release of UA (95.67% in 48 h) compared to gellan gum NPs (96.12% in 8 h). In cytotoxicity studies, UA loaded HAG NPs exhibited an enormous cytotoxic potential against A549 cancer cells. In the in vivo bio-distribution study, using albino rat model the free UA concentration was found 7.09 ±â€¯0.9%, 2.7 ±â€¯1.5%, 7.5 ±â€¯2.1, 9.2 ±â€¯2%, and 6.25 ±â€¯1.3% post two hours of intravenous administration, however, in the case of UA loaded HAG NPs the obtained level was 4.1 ±â€¯1.10, 7.7 ±â€¯1.30%, 2.21 ±â€¯0.29%, 1.85 ±â€¯0.25%, 2.2 ±â€¯0.78%, 2.9 ±â€¯1.21% respectively, in heart, lung, liver, spleen, intestine and kidney. The overall anticancer study and result of internalization deciphered the higher anticancer potential of UA loaded HAG NPs.

[Evaluation of clinical efficacy of serotonin adipate in treatment and prevention of enteral insufficiency syndrome at generalized peritonitis]. / Otsenka klinicheskoi effektivnosti preparata serotonina adipinat v lechenii i profilaktike sindroma enteral'noi nedostatochnosti pri rasprostranennom peritonite.

AIM: To substantiate pathogenetic expediency and to evaluate the clinical efficacy of the drug use serotonin adipate (dinaton) in a complex correction enteral insufficiency syndrome (EIN) in patients with generalized peritonitis (GP). MATERIAL AND METHODS: The comparative analysis of results of treatment of 182 patients with GP, which in principle approach to EIN correction in the postoperative period were divided into two groups. Group I consisted of 92 patients who received standard intensive therapy using conventional methods of stimulation of intestinal peristalsis. Group II consisted of 90 patients on a background of standard treatment was carried further pharmacological stimulation of intestinal motility drug serotonin adipate (dinaton). The research program included an assessment of clinical parameters intestinal motility recovery, evaluation of the severity of the patients on a scale of APACHE II, determining the blood levels of serotonin and the level of the main biomarkers of systemic inflammatory response (SIR), the study of blood flow in the vessels of splanchnic bed, the measurement of intra-abdominal pressure with the calculation of intraperitoneal perfusion pressure. RESULTS: It is found that the development and progression of abdominal inflammation is accompanied by a sharp decrease in blood serotonin level is in phase III GP decreases 4.7 times compared to the control value. It is shown that using of serotonin adipate (dinaton) in treatment of patients with GP promotes early recovery of intestinal motility and the resolution of EIN, the elimination of intra-abdominal hypertension and disorders of splanchnic blood flow, as well as the rapid regression of the manifestations of the SIR and endotoxemia. Postoperative mortality in group I patients was 28.3% in group II - 20.0%. CONCLUSION: The inclusion of serotonin adipate (dinaton) in the complex corrective therapy standard in the postoperative period in GP patients is pathogenetically justified, as it promotes early restoration of motor activity of the gastrointestinal tract, the elimination of intestinal paresis and resolution of EIN, which leads to an improvement of results of surgical treatment of this patients.

[EFFICIENCY OF SEROTONIN ADIPINATE IN INTESTINAL DYSFUNCTION IN PATIENTS AFTER COLORECTAL OPERATIONS].

We performed a retrospective analysis of case histories of acute colonic obstruction due to colon cancer A total of 291 patients were divided on two groups: 1--a control group (patients presenting risk of developing intestinal dysfunction with 'basic' therapy, n = 123); 2--the comparison group (n = 57) represented patients who were taken to optimize the post-operative period with the inclusion in the scheme of the basic treatment of serotonin adipinate. The use of serotonin adipinatein treatment of intestinal dysfunction allows fully restore bowel motility to 3rd day.

Phthalate and di-(2-ethylhexyl) adipate (DEHA) intake by German infants based on the results of a duplicate diet study and biomonitoring data (INES 2).

Phthalates as well as di-(2-ethylhexyl) adipate (DEHA) are used as plasticizers in diverse applications and are of toxicological concern. The study was conducted with a study population of 25 German subjects aged between 15 and 21 months. Overall, 16 phthalates and DEHA were measured by gas chromatography-mass spectrometry in a total of 171 duplicate diet samples collected over 7 consecutive days, and 20 phthalate metabolites were analyzed in the urine samples collected over 7 consecutive days using a liquid chromatography-tandem mass spectrometry method. The median "high" daily dietary intake based on 95th percentiles was 4.66 µg/kg b.w. for di-2-ethylhexyl phthalate (DEHP), 1.03 µg/kg b.w. for di-isobutyl phthalate (DiBP), and 0.70 µg/kg b.w. for di-n-butyl phthalate (DnBP), and 1.0 µg/kg b.w. for DEHA. The "high" daily total intake from biomonitoring data was 4.9 µg/kg b.w. for DEHP, 2.2 µg/kg b.w. for DnBP, 3.9 µg/kg b.w. for DiBP, and 2.6 µg/kg b.w. for di-isononyl phthalate. The comparison of the two intake estimates indicates that the dominant intake source of DEHP was food ingestion, whereas other sources considerably contributed to the total intake of other phthalates. Using our "high" intake scenario, none of the analyzed phthalates reached the recommended tolerable daily intake levels.

An injectable oxidated hyaluronic acid/adipic acid dihydrazide hydrogel as a vitreous substitute.

Vitrectomy is a common procedure for treating ocular-related diseases. The surgery involves removing the vitreous humor from the center of the eye, and vitreous substitutes are needed to replace the vitreous humor after vitrectomy. In the present study, we developed a colorless, transparent and injectable hydrogel with appropriate refractive index as a vitreous substitute. The hydrogel is formed by oxidated hyaluronic acid (oxi-HA) cross-linked with adipic acid dihydrazide (ADH). Hyaluronic acid (HA) was oxidized by sodium periodate to create aldehyde functional groups, which could be cross-linked by ADH. The refractive index of this hydrogel ranged between 1.3420 and 1.3442, which is quite similar to human vitreous humor (1.3345). The degradation tests demonstrated that the hydrogel could maintain the gel matrix over 35 days, depending on the ADH concentration. In addition, the cytotoxicity was evaluated on retina pigmented epithelium (RPE) cells cultivated following the ISO standard (tests for in vitro cytotoxicity), and the hydrogel was found to be non-toxic. In a preliminary animal study, the oxi-HA/ADH hydrogel was injected into the vitreous cavity of rabbit eyes. The evaluations of slit-lamp observation, intraocular pressure, cornea thickness and histological examination showed no significant abnormal biological reactions for 3 weeks. This study suggests that the injectable oxi-HA/ADH hydrogel should be a potential vitreous substitute.

Collaborative work on evaluation of ovarian toxicity. 9) Effects of 2- or 4-week repeated dose studies and fertility study of di(2-ethylhexyl)adipate (DEHA) in female rats.

The present study was designed to confirm whether or not the ovarian toxicity of di(2-ethylhexyl)adipate (DEHA), which is known to have effects on female fertility, could be evaluated by the new method of histopathological examination of the ovaries in repeated dose toxicity. DEHA was orally administered to Crl:CD(SD) female rats at the doses of 0, 200, 1,000 and 2,000 mg/kg for 2 or 4 weeks in repeated dose toxicity study and for 2 weeks before mating, throughout mating and until Gestation Days 7 in female fertility. In the repeated dose toxicity studies, increase in atresia of large follicle, decrease in currently formed corpus luteum and follicular cyst were observed in the 1,000 mg/kg and above groups, suggesting that DEHA disturbed ovulation and large follicle growth. In the fertility study, a significant increase in mean estrus cycle length and post-implantation loss rate were observed in the 1,000 mg/kg and above groups, and a significant decrease in implantation rate and number of live embryos and a significant increase in pre-implantation loss rate were observed in the 2,000 mg/kg group. The histopathological changes of ovary observed in the repeated dose toxicity studies were correlated with the result that DEHA affected the estrus cycle in the female fertility study. In conclusion, a 2-week administration period is sufficient for detection of the ovarian toxicities following treatment with DEHA by new histopathological examination of the ovaries.

[Bone targeting of antitumor conjugate phenamine acid aryolysine-hexanedioic acid bridge grafting-bisphosphonates].

OBJECTIVE: To develop a new bone targeting antitumor therapy system which uses diphosphonate and bone-seeking nuclide to enhance the coordinated effects of chemotherapy and radiotherapy on bones, and to validate the targeting of the new therapy system in vitro and in vivo. METHODS: Phenamine acid caryolysine was connected to bisphosphonates, and then combined with radioactive nuclide 153Sm to establish a new bone targeting chemotherapeutic and radioactive drug for bone cancers. The targeting of this new therapy system was validated by hydroxyapatite crystal absorbing test and body distribution in vivo methods. RESULTS: The optical spectrum of the phenamine acid caryolysine-bisphosphonates conjugate detected by the nuclear magnetic resonance was consistent with the standard structure of synthesized drugs. The conjugate had good absorbability to hydroxyapatite crystals. The body distribution of the conjugate showed higher radiocounting in bones than in other tissues. CONCLUSION: The conjugate has a structure that is consistent with the active compound targeting bone tumors.

Quantitative acylcarnitine profiling in peripheral blood mononuclear cells using in vitro loading with palmitic and 2-oxoadipic acids: biochemical confirmation of fatty acid oxidation and organic acid disorders.

Organic acid (OAD) and fatty acid oxidation disorders (FAOD) are inborn errors of metabolism often presenting with life-threatening metabolic decompensation followed by (irreversible) organ failure, and even death during catabolic state. Most of these diseases are considered as treatable, and metabolic decompensations can be avoided by early diagnosis and start of therapy. Confirmation of suspected diagnosis currently relies on enzymatic and mutation analyses and in vitro loading of palmitic acid in human skin fibroblast cultures. Furthermore, in some cases potentially life-threatening in vivo loading or fasting tests are still performed. In this study, we established a standardized in vitro loading test in peripheral blood mononuclear cells (PBMC) that allows reliable biochemical confirmation of a suspected diagnosis within 1 week. Patients with confirmed diagnosis of short-, medium-, very-long-chain, and long-chain 3-hydroxyacyl-CoA dehydrogenase deficiencies, methylmalonic, propionic, isovaleric acidurias, and glutaric aciduria type I were included in the study. PBMC, isolated from heparinized venous blood samples of these individuals were incubated for 5 days with palmitic acid or 2-oxoadipic acid (glutaric aciduria type I), respectively, and quantitative acylcarnitine profiling was subsequently performed in supernatants using electrospray ionization tandem mass spectrometry. All patients were clearly identified, including those with mild biochemical phenotypes who, in particular, are at risk to be missed under balanced metabolic conditions. In glutaric aciduria type I, the same results were also obtained using lymphoblasts. In conclusion, our assay allows biochemical confirmation of a number of FAOD and OAD and could easily be implemented into the confirmatory diagnostic work-up.

[Effect of serotonin-adipinate on the ultrastructure and synthetic activity of endotheliocytes of microhemocirculatory bed of human brain (autopsy material, early autopsies)]. / Vozdeistvie serotonina adipinata na ul'trastrukturu u sinteticheskuiu aktivnost' éndoteleotsitov mikrogemotsirkuliarnogo rusla golovnogo mozga cheloveka.

Electron-microscopic and radioautographic investigation showed the ability of brain capillary endotheliocytes for retaining functional activity in total ishemia. Administration of serotonin adipinate in an early postoperative period supports intracellular processes and plays a protective role.

[The treatment of critical lower limb ischemia using serotonin]. / Lechenie kriticheskoi ishemii nizhnikh konechnostei s ispol'zovaniem serotonina.

This paper describes the first experience with conservative treatment of 30 men suffering from critical lower limb ischemia using serotonin as an agent which may return endogenous vasomotility to normal and eliminate tissue hypoxia. Good and satisfactory results were obtained in 76% of patients (43 and 33% respectively). Deterioration followed by amputation was marked in 7% of patients. The removal or considerable abatement of rest pains was recorded in 86% (53 and 33% respectively) of cases. Amputation was performed in 10% of patients. It is recommended that serotonin adipinate should be included into multiraodality therapy of atherosclerotic critical lower limb ischemia. The use of the treatment method suggested will allow to improve the treatment results, to reduce the number and level of amputations, and to minimize the percentage of disability.

Role of serotoninergic structures in coordination of electric activity in the gastroduodenal complex.

Myoelectric activity in various portions of the stomach and duodenum in normal and after vagotomy and intravenous injection of serotonin adipinate was studied in acute experiments on cats. Vagotomy disturbed coordination of myoelectric activity in the stomach and duodenum. Intravenous injection of serotonin adipinate restored coordination and increased myoelectric activity in the stomach, particularly, in the pylorus. Under these conditions the pacemaker of myoelectric activity in the gastric corpus and autonomic regulation of the duodenum were preserved. Intravenous injection of serotonin adipinate most significantly changed myoelectric activity in the pylorus. Myoelectric activity in the cardia increased after vagotomy against the background of serotonin adipinate. Our findings suggest that serotoninergic structures maintain functional heterogeneity of digestive organs and coordinate their interrelationships.

Dietary adipic acid reduces ammonia emission from swine excreta.

Adipic acid is only partially catabolized when it is fed to animals, and a portion of it is excreted in urine. The excreted portion may lower urinary pH and, as a result, ammonia emission. The present study tested this hypothesis. In Exp. 1, nursery pigs (n = 14) were fed (for a period of 7 d) either a standard nursery diet or the same diet supplemented with 1% adipic acid to assess effects on urinary pH (collected on d 5 or 6) and in vitro ammonia emission from the collected urine samples that were mixed with control feces. In Exp. 2, grower pigs housed 10 each in one of two chambers were fed a control diet or the same diet supplemented with 1% adipic acid. Ventilated air was quantified and analyzed for ammonia using Fourier transform infrared spectroscopy to determine the effects of feeding 1% adipic acid on ammonia emission. The results from Exp. 1 showed that adipic acid strongly reduced urinary pH (from 7.7 to 5.5, P < 0.05). In vitro ammonia emission from these urine samples was significantly reduced at all the time points evaluated (1, 3, 18, and 46 h with reductions of 94, 93, 70, and 39%, respectively, P < 0.05). Experiment 2 showed that adipic acid supplementation reduced ammonia emission by 25% (P < 0.05), which corresponded to the predicted reduction in ammonia emission based on the reduction in manure pH observed. In conclusion, feeding adipic acid lowers urinary pH and reduces ammonia emission. The reduction in ammonia emission, though, does not correspond to the reduction in urinary pH but corresponds to the reduction in fecal pH as a result of mixing the urine and feces, in which feces act as a strong buffer.

Isolated adipic aciduria.

Adipic acid can appear, in combination with other dicarboxylic acids, in the urine of patients in a number of underlying metabolic diseases. A child with seizures and mental retardation of unknown etiology who was found to have elevated isolated adipic aciduria on investigation for metabolic diseases is reported. A dietary artifact was suspected, and the adipic aciduria resolved after the child was kept on a specific restricted diet for 3 days. This is the third report of isolated adipic aciduria secondary to food. Findings confirm the previous reports of dietary origin of isolated adipic aciduria and should alert clinicians to such artifact before committing patients to unnecessary treatments.

[Use of serotonin for treatment of patients with diabetic angiopathies]. / Primenenie serotonina dlia lecheniia bol'nykh s diabeticheskimi angiopatiiami.

In 26 patients (female 11, males--15) with diabetic angiopathy (necrosis of the foot or toes) local measurement of pO2 by transcutaneous mode was carried out before and during introduction of serotonine adipinate. Simultaneously with evaluation of pO2 in 14 patients electromyography (EMG) was made. Serotonine adipinate was introduced intravenously with a speed of 5.0-10.0 mg per hour. In this group mean age of the patients made up 57.4 years, mean values of initial pO2 25.6 mm Hg, and mean indices of pO2 at the moment of serotonine treatment 38.5 mm Hg. Average increase of pO2 during serotonine infusion was registered as 50.4% in comparison with mean initial data. In all 14 patients, who underwent simultaneous measurement of pO2 and registration of EMG, reinforcement of electrical activity of smooth muscles (SM) of various intensity and increase in tissue pO2 were detected. Improvement of microcirculation (increase of transcutaneous pO2, reinforcement of electrical activity of SM as a result of serotonine introduction) evidences that functional and morphological changes of serotonine receptors of SM are involead in genesis of diabetic angiopathy. The data obtained showed that in diabetes mellitus, besides insulin deficiency, insufficiency of such an endogenous factor as serotonine duclops. Thus, serotonin should be used for prophylaxis and treatment of diabetic angiopathies.

[Use of serotonin adipinate in combined therapy of hemodynamic disorders in acute poisoning by psychopharmacological preparations]. / Primenenie serotonina adipinata v kompleksnoi terapii narushenii gemodinamiki pri ostrykh otravleniiakh psikhofarmakologicheskimi preparatami.

Serotonin adipinate in a dose of 40-60 mg was injected by drip infusion to 30 patients with acute poisoning by psychotropic drugs for correcting circulatory disorders. The drug corrected hemodynamic disorders in the lesser and greater circulation by increasing the stroke volume and normalizing total peripheral vascular resistance and central blood volume.

Developmental toxicity of dibasic esters by inhalation in the rat.

Dibasic esters (DBE) are a mixture of 3 carboxylic acid esters which are used in the paint and coatings industry. In this study, groups of pregnant Crl:CD BR rats were exposed to either 0.16, 0.4, or 1.0 mg DBE/L by inhalation for 6 hr/day from Days 7 through 16 of gestation (day in which copulation plug was detected was designated Day 1G). A control group of chambered pregnant rats was exposed simultaneously to air only. All female rats were euthanized on Day 21G and the fetuses were examined. A suppression of both food consumption and the rate of body weight gain was seen in the 0.4 and 1.0 mg/L groups during the first 6 exposure days. Staining on the fur and perineal area was seen in rats exposed to 1.0 mg/L and liver weight decreases, although not statistically significant, occurred in the 2 high exposure groups. None of the reproductive parameters were altered in any of the groups and no fetal effects were detected. DBE is not a developmental toxin in the rat following inhalation exposures as high as 1.0 mg DBE/L during the period of organogenesis.