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Continuous delivery of neurotrophin 3 by gene therapy has a neuroprotective effect in experimental models of diabetic and acrylamide neuropathies.

Neurotrophic factors (NFs) are promising agents for the treatment of peripheral neuropathies such as diabetic neuropathy. However, the value of treatment with recombinant NF is limited by the short half-lives of these molecules, which reduces efficiency, and by their potential toxicity. We explored the use of intramuscular injection of a recombinant adenovirus encoding NT-3 (AdNT-3) to deliver sustained low doses of NT-3. We assessed its effect in two rat models: streptozotocin (STZ)-induced diabetes, a model of early diabetic neuropathy characterized by demyelination, and acrylamide experimental neuropathy, a model of diffuse axonal neuropathy which, like late-onset human diabetic neuropathy, results in a diffuse sensorimotor neuropathy with dysautonomy. Treatment of STZ-diabetic rats with AdNT-3 partially prevented the slowing of motor and sensory nerve conduction velocities (p < 0.01 and p < 0.0001, respectively). Treatment with AdNT-3 of acrylamide-intoxicated rats prevented the slowing of motor and nerve conduction velocities (p < 0.001 and p < 0.0001, respectively) and the decrease in amplitude of compound muscle potentials (p < 0.0001), an index of denervation. Acrylamide-intoxicated rats treated with NT-3 had higher than control levels of muscle choline acetyltransferase activity (p < 0.05), suggesting greater muscle innervation. In addition, treatment of acrylamide-intoxicated rats with AdNT-3 significantly improved behavioral test results. Treatment with AdNT-3 was well tolerated with minimal muscle inflammation and no detectable general side effects. Therefore, our results suggest that NT-3 delivery by adenovirus-based gene therapy is a promising strategy for the prevention of both early diabetic neuropathy and axonal neuropathies, especially late axonal diabetic neuropathy.

Acrylamides↗

The number and mean volume of neurons in the cerebral cortex of rats intoxicated with acrylamide.

Acrylamide was given in an accumulated dose of 500 mg/kg to rats by intraperitoneal injection in two different dosing schedules: 50 mg/kg twice a week for 5 weeks and 33.3 mg/kg twice a week for 7.5 weeks. The effect of acrylamide intoxication on the neurons in the cerebral cortex of the rat was studied using unbiased stereological methods. A reduction of brain weight of 8% was seen in both the intoxicated groups. The volume of neocortex was significantly decreased in the experimental groups, but the density of neurons was increased resulting in an unchanged total number of neurons. The mean volume of neurons in neocortex was significantly decreased in both acrylamide intoxicated groups. There was no difference between the two different intoxication schedules. The possibility that acrylamide causes neuronal death and the effect of eventual differential cellular sensitivity is discussed.

Acrylamides↗

The acute effects of acrylamide on astrocyte functions.

We assessed biochemical endpoints indicative of acute toxicity in neonatal rat primary astrocyte cultures exposed to acrylamide. Metallothionein (MT), glutamine synthetase (GS), glutamate/aspartate transporter (GLAST), and taurine transporter (tau-T) mRNA expression levels as well as cell volume were determined in astrocytes acutely treated with 0.1 and 1.0 mM acrylamide. Statistically significant changes in acrylamide treated astrocytes were noted for GS (0.1 mM) and GLAST (1.0 mM) mRNA expression levels. All other measurements were insignificant in comparison with controls, suggesting that astrocytic function is minimally compromised even at exceedingly high levels of acute acrylamide exposure.

Acrylamide↗

Thermosensitive nanocontainers prepared from poly(N-isopropylacrylamide-co-N-(hydroxylmethyl) acrylamide)-g-poly(lactide).

Thermally-responsive graft copolymer of poly(N-isopropylacrylamide-co-N-(hydroxylmethyl)acrylamide)-g-poly(lactide) was synthesized by ring-opening polymerization of D,L-lactide (LA). The polymerization was initiated by the hydroxy group of poly(N-isopropyl acrylamide-co-N-(hydroxylmethyl) acrylamide), using stannous octoate as catalyst. The resulting polymer was temperature-sensitive and the lower critical solution temperature (LCST) was affected by their composition. The chemical structure and physical properties of the grafted copolymers were investigated by various methods. Nanocontainers formed from the self-assembly of poly(N-isopropylacrylamide-co-N-(hydroxylmethyl) acrylamide)-g-poly(lactide) were characterized by transmission electron microscopy (TEM), and a spherical structure was observed. Dynamic light scattering (DLS) results indicate that the particle size increased with the increase of polylactide content in the copolymer. The properties of this polymer are interesting for both industrial application and fundamental research. In particular it will combine a spatial specificity in a passive manner and a temperature-responsive active targeting mechanism for drug delivery system.

Acrylamides↗

Development and single-laboratory validation of a reversed-phase liquid chromatography-electrospray-tandem mass spectrometry method for identification and determination of acrylamide in foods.

A rapid and accurate method using reversed-phase liquid chromatography-tandem mass spectrometry interfaced with electrospray was developed for determination of acrylamide in cooked food samples. A simplified sample treatment procedure using an extraction step with acidified water without cleanup was developed. A C18 column with an aqueous formic acid-methanol mixture as the mobile phase was used under isocratic conditions. The method was validated in-house for robustness, limits of detection (LOD) and quantitation (LOQ), linearity, recovery, and accuracy both on standard and baked-product and potato flour matrixes. Good results in the low ppb level were obtained for LOD (< 15 microg/kg) and LOQ (< 25 microg/kg) of acrylamide in samples. Excellent linearity (r2 = 0.999-1.000) was established over 2 orders of magnitude by performing statistical tests. The absence of both constant and proportional systematic errors demonstrated good method accuracy. Excellent results were obtained for intraday repeatability (RSD < 1.5%) and between-day precision (RSD < 5%). Extraction recoveries from food products were calculated in the 97 +/- 3-99 +/- 2% (n = 6) range with a labeled internal standard (13C3-acrylamide). The applicability of the method to determination of acrylamide in cooked food products was demonstrated.

Acrylamides↗

Quantitation of acrylamide in food products by liquid chromatography/mass spectrometry.

A simple and inexpensive liquid chromatography/mass spectrometry (LC/MS) method was developed for the quantitation of acrylamide in various food products. The method involved spiking the isotope-substituted internal standard (1-C13 acrylamide) onto 6.00 g of the food product, adding 40 mL distilled/deionized water, and heating at 65 degrees C for 30 min. Afterwards, 10 mL ethylene dichloride was added and the mixture was homogenized for 30 s and centrifuged at 2700 x g for 30 min, and then 8 g supernatant was extracted with 10, 5, and 5 mL portions of ethyl acetate. The extracts were combined, dried with sodium sulfate, and concentrated to 100-200 microL. Acrylamide was determined by analysis of the final extract on a single quadrupole, bench-top mass spectrometer with electrospray ionization, using a 2 mm id C18 column and monitoring m/z = 72 (acrylamide) and m/z = 73 (internal standard). For difficult food matrixes, such as coffee and cocoa, a solid-phase extraction cleanup step was incorporated to improve both chromatography and column lifetime. The method had a limit of quantitation of 10 ppb, and coefficients of determination (r2) for calibration curves were typically better than 0.998. Acceptable spike recovery results were achieved in 11 different food matrixes. Precision in potato chip analyses was 5-8% (relative standard deviation). This method provides an LC/MS alternative to the current LC/MS/MS methods and derivatization gas chromatography/mass spectrometry methods, and is applicable to difficult food products such as coffee, cocoa, and high-salt foods.

Acetates↗

[Toxicity study for acrylamide].

Acrylamide is one of the most important chemical materials, which has been widely used in industry. It is one kind of many contaminants in occupational environments. However, it has been reported that starchy foods cooked at high temperature contain a certain amount of acrylamide. The main acrylamide exposure route for non-professional population is intaking of high-temperature starchy foods. In this review, the studies for tumorogenesity, neurotoxicity, reproductive and developmental toxicity possibly caused by acrylamide were reported.

Acrylamide↗

Position sensitivity of de-efferented muscle spindles in experimental acrylamide neuropathy.

An experimental neuropathy was induced in cats by injections of acrylamide (7.5, 15 or 30 mg/kg/day) for 2 to 10 days. The responses of primary and secondary endings of soleus muscle spindles to stretch were evaluated and correlated with the appearance of ataxia and incoordinated motor movements. Animals that received 15 or 30 mg/kg/day became ataxic and demonstrated poor motor coordination on the 7th or 4th day, respectively. At these times, both primary and secondary endings of muscle spindles had elevated thresholds and diminished discharge frequencies. Continued acrylamide administration resulted in exacerbation of the clinical symptoms and further attenuation of spindle responses. The discontinuation of acrylamide was followed by slow recovery. Only those cats which received a total dose of 75 mg/kg or less remained asymptomatic and had normal spindle function. The coincidence of onset of motor coordination deficits and spindle dysfunction, coupled with a lack of demonstrable motor defect at the same time, suggests that the initial clinical features of acrylamide neuropathy may be partly the consequence of impaired spindle function.

Acrylamides↗

Effect of forced running on rat skeletal muscle with acrylamide neuropathy.

This study was performed to evaluate the effect of prolonged forced running on rat lower limb muscles with acrylamide neuropathy. Twenty-four 4-week-old rats were divided into three groups of eight rats. Acrylamide was given to two groups of sixteen rats to induce mild paralysis. Eight rats with acrylamide injections were forced to run 3,200 m/day on a treadmill for five weeks. Running activities slowed the rate of body weight gain and aggravated paralysis. Although the wet weight of tibialis anterior (TA) and extensor digitorum longus (EDL) muscles was reduced by running, that of soleus (SOL) muscles was unchanged. The ratio of their weight to their body weight (BW) remained constant regardless of exercise. Protein content (PC) of muscles was not altered by exercise, either. We postulated that exercise-induced worsening of paralysis in acrylamide neuropathy rats was not caused by muscle pathology. Deterioration of neuropathic condition due to exercise was suggested.

Acrylamides↗

[Interaction of methylmethacrylate and acrylamide with the microsomal oxidation system of the rat liver].

A complex, exhibiting the maximal absorption at 385-388 nm and minimal at 414-420 nm, was formed after interaction between cytochrome P-450 and methylmethacrylate and acrylamide at 5 mM concentration. Acrylamide at concentrations higher than 6 mM induced the formation of modified type 11 differential spectra with maxima at 418 nm and minima at 385 nm. Methylmethacrylate and acrylamide stimulated utilization of NADPH and inhibited hydroxylation of 3,4-benzpyrene. Methylmethacrylate inhibited also N-demethylation of aminopyrine. After treatment of rats with the monomers activities of cytochromes P-450, b5 and NADPH-cytochrome c reductase were unaltered. Methylmethacrylate and acrylamide appear to interact with the microsomal oxidative system.

Acrylamides↗

Polyneuropathies and CNS protein metabolism. III. Changes in protein synthesis rate induced by acrylamide intoxication.

A defect in neuronal protein metabolism has been proposed as one of the primary, molecular events underlying the development of polyneuropathies of the dying back type. Using acrylamide-intoxication as an experimental model to study these polyneuropathies, changes in leucine-incorporation into proteins disability. Proteinsynthesis rates were determined in vivo using flooding concentrations of [1-14C]valine as the precusor. Under conditions of acute and of chronic intoxication, a decrease in synthesis rate was measured preceding the loss of functional ability. Similar changes in protein synthesis rate were observed in peripheral tissues such as heart muscle and liver showing the general toxicity of acrylamide. Methylene bisacrylamide, that was used to discriminate between the neurotoxic action of acrylamide and its systemic effects, interfered with protein synthesis rates in a comparable way. No change in protein synthesis rate was observed under in vitro conditions suggesting that the interference of acrylamide with the synthetic machinery for protein synthesis in vivo is mediated by one or more as yet unknown indirect factors.

Acrylamides↗

Mesotheliomas of tunica vaginalis testis of Fischer 344 (F344) rats treated with acrylamide: a light and electron microscopy study.

The incidence of spontaneous mesotheliomas of the tunica vaginalis testis, documented to be 1.3% in Fischer 344 rats can be increased by long term exposure to acrylamide. The light microscopic and electron microscopic features of tumors developing spontaneously in control animals were compared with those that developed in acrylamide treated animals to determine whether the tumors in the experimental group have any distinctive characteristics. The experimental group comprised three subgroups, i.e. animals exposed to acrylamide in three doses 0.1, 0.5 or 2.0 mg/kg/day over a period of 104 weeks. A total of 35 tumors were examined--7 in control group, 9 in low-dose group, 8 in mid-dose group and 11 in the high-dose group. Most tumors measured less than 1 cm in diameter in aggregate and approximately one-third of all tumors were of microscopic size, less than 5 mm in diameter. All tumors were classified as epithelial mesotheliomas. Three distinct microscopic patterns were recognized (papillary, tubular or solid). All tumors had papillary features, and focally showed some tubular or solid components. No ultrastructural differences were found between tumors of the four groups of animals. It was concluded that the tumors that developed in the tunica vaginalis testis of acrylamide treated animals do not differ morphologically from those that developed spontaneously in control animals. Cellular uniformity, small size of lesions and the absence of perotoneal seeding and metastasis suggests that the mesotheliomas of tunica vaginalis testis of Fischer 344 rats in this study were benign and equivalent to human benign mesothelioma of tunica vaginalis testis or adenomatoid tumors.

Acrylamide↗

Electrophoresis of proteins and nucleic acids on acrylamide-agarose gels lacking covalent crosslinking.

The preparation of acrylamide-agarose gels lacking covalent crosslinking with methylenebisacrylamide is described. These hybrid gels melt at 85 degrees C and, consequently, allow quantitative analysis of tritium-labeled protein after electrophoresis. Recovery of tritium-labeled ribonucleic acids extracted from hybrid gels is 20 to 25% greater than from standard acrylamide-methylenebisacrylamide gels. Standard curves of electrophoretic mobilities as a function of molecular weights of dissociated proteins and ribonucleic acids are compared for acrylamide-agarose gels and acrylamide-methylenebisacrylamide gels.

Electrophoresis, Agar Gel↗

Polyacrylamide gel electrophoresis: reaction of acrylamide at alkaline pH with buffer components and proteins.

The chemical reaction of monomeric acrylamide with primary, secondary, and tertiary amines, used as buffer components in polyacrylamide gel electrophoresis systems, was investigated in the basic pH range. Adduct formation proceeded for several minutes up to weeks, depending on the reactivity of the amino groups. A pH shift in the reaction mixture due to an altered pK value of the reaction product was observed. However, a few primary amines (tris(hydroxymethyl)aminomethane, 2-amino-2-methyl-1,3-propanediol) and secondary amines 3-([2-hydroxy-1,1-bis(hydroxymethyl)ethyl]amino)-1-propanesulfonic acid, 3-(dimethyl(hydroxymethyl)methylamino)-2-hydroxypropanesulfonic acid) showed negligible shifts of pH. They are, therefore, useful as components in the polymerization mixture; whereas some tertiary amines showing complete pH stability as well (e.g., triethanolamine) are not suitable, as they acted as accelerators of gel polymerization. Acrylamide can also covalently bind to proteins by reacting with the epsilon-amino group of lysine residues, especially. Bovine serum albumin, having an acidic isoelectric point, and the basic protein cytochrome c were treated with different acrylamide concentrations at alkaline pH yielding modified protein molecules with altered electrophoretic mobilities in different polyacrylamide gel electrophoresis systems. This reaction gave rise to artifacts in alkaline polyacrylamide gels and isoelectric focusing systems when residual acrylamide monomers were still present in the gel matrix after the polymerization process ceased.

Amines↗

Hydrogen bonding and the conformations of poly(alkyl acrylamides).

The conformations of poly(alkyl acrylamide) oligomers in nonpolar solvents were studied using molecular dynamics techniques. Poly(methyl acrylamide) was found to collapse to a globule-like conformation at low temperatures; however, excluded volume effects inhibited the collapse of poly(octadecyl acrylamide). A high density of structured units, characterized by a trans-gauche-trans-trans-gauche-trans torsional sequence along the backbone, was noted in all simulations. Such units were found to create a particularly stable set of intramolecular hydrogen bonds. An oligomer constructed with these stable units was found to have significantly lower minimized energy than both the all-trans and the helical backbone conformations. The constructed conformation had lower Coulomb energy (more hydrogen bonds) than the all-trans conformation and lower dihedral energy (less backbone distortion) than the helical conformation. The propensity for poly(octadecyl acrylamide) to form hydrogen bonds introduced significant disorder into the orientation of the alkyl side chains. This disorder would inhibit crystallization and restrict the ability of such polymers to form epitaxial seeds for nucleating paraffin crystals.

Journal Article↗

Cadmium sulfide and cadmium selenide/cadmium sulfide nanoparticles stabilized in water with poly(cysteine acrylamide).

Cysteine acrylamide (N-acryloyl L-cysteine) stabilizes CdS nanoparticles as the particles form in aqueous dispersions. Cysteine acrylamide also exchanges for citrate on the surfaces of CdSe and core/shell CdSe/CdS nanoparticles to provide greater stability. Heating of the nanoparticle dispersions polymerizes the cysteine acrylamide on the surface to form a more efficient polydentate stabilizer. The polymer-coated nanoparticle dispersions are colloidally stable even after removal of low molecular weight solutes by dialysis. Emission quantum yields of the polymer-coated CdSe and CdSe/CdS samples were 0.9% and 2.6%, respectively, after aging of the samples in light. CdSe/CdS coated with poly(cysteine acrylamide) is colloidally stable for at least two years in the dark at 5 degrees C.

Acrylic Resins↗

Toxicity of polyacrylamide and acrylamide monomer.

The purpose of this paper is to present information gathered regarding, in general, the physical characteristics, and, in particular, the possible toxic nature of polyacrylamide A short discussion of the properties and toxicity of the acrylamide monomer is also included. The United States government, specifically the Food and Drug Administration and the Environmental Protection Agency, already regulates several uses of polyacrylamide; criteria and standards have been established based on numerous toxicological studies of both polyacrylamide and acrylamide. These studies are reviewed and summarized. The regulations generally restrict both the amount of residual acrylamide monomer in the polyacrylamide and the amount of polymer that may be used in the specified application. By imposing this type of restriction, a maximum limit on the amount of acrylamide in contact with food or drinking water can be indirectly achieved.

Acrylic Resins↗

Two-dimensional acrylamide gel electrophoresis of cancer-patient serum proteins.

Sera from normal volunteers, patients with a variety of non-neoplastic diseases and patients with malignant or benign tumors were examined by two-dimensional acrylamide gel electrophoresis. In this technique the serum is first separated in an acrylamide gel column followed by a second electrophoresis at right angles to the first separation in a continuous concave 2 to 30 percent gradient acrylamide gel slab. The stained two-dimensional gel slab appears as a "fingerprint" pattern or "map" of the separated serum proteins. Both qualitative and quantitative differences in the fingerprint patterns of cancer-patient sera were observed. The quantitative alterations did not appear specifically associated with malignant tumors. However, several qualitative differences were detected, of which some may represent markers of malignancy as they were not observed in the normal, abnormal and benign tumors control sera. At least one of the abnormal protein stained spots, a prealbumin, appears to be restricted and related in some way to cancer of the lymphoreticular system. These data, although limited, support earlier indications that two-dimensional acrylamide gel electrophoresis offers a new and powerful tool to the cancer scientist for the detection of alterations in cancer-patient sera. A discussion of its possible use to examine other biological fluid and tumor extracts from the cancer patient is presented.

Blood Protein Electrophoresis↗