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PubMedDate not supplied

[Imbalance of plasma amino acids in patients with autism and subjects with attention deficit/hyperactivity disorder].

INTRODUCTION: Plasma and brain amino acids are influenced by dietary intake. Alterations of plasma amino acid concentrations have been reported in neuropsychiatric disorders. OBJECTIVE: To analyse the plasma amino acid values in subject diagnosed with autism, with attention deficit/hyperactivity disorder (ADHD), and healthy subjects as controls. PATIENTS AND METHODS: Forty subjects affected by autism, 11 with ADHD and 41 healthy subjects (age range 3 18 years old) were included in this study. Peripheral venous blood was obtained in fasting condition, collected in EDTA tubes and centrifuged. Plasma was de proteinised with sulfosalicylic acid. Amino acids were analysed by ion exchange liquid chromatography with an LKB amino acid analyser with sodium citrate elution system and ninhydrin reaction. Results were expressed as mmol/L. RESULTS AND CONCLUSIONS: In both disorders a diminution of phenylalanine and glutamine plasma concentrations was observed beside an increase of glycine. Lysine appeared increased only in autistic subjects. These alterations produce an imbalance with the rest of plasma amino acids competing at the brain blood barrier by the same transport system thus causing alterations in the metabolism and/or transport of amino acids to the brain, altering CNS functions. The phenylalanine decreasing, beside glycine increasing appear to support the hypothesis of a disorder in the inhibitory neurotransmission system, especially in ADHD. The diminution of phenylalanine and the increasing of lysine in autism are suggestive that these two amino acids are metabolically related.

Adolescent↗

PubMed1975

More sensitive automated detection of polyamines in physiological fluids and tissue extracts with omicron-phthalaldehyde.

Omicron-Phthalaldehyde reacts with primary amines to form an intensely blue-fluorescing product. Replacing ninhydrin with this compound in our automated high-pressure liquid-chromatographic technique for determination of polyamines, we achieved a 6- to 10-fold increase in sensitivity. Linearity and reproducibility are excellent, and the method is shown to be applicable to analysis of physiological fluids and of tissue extracts.

Aldehydes↗

PubMed2002

Extension of the color suite available for chemical enhancement of fingerprints in blood.

The use of ortho- and para-phenylenediamine (OPD & PPD respectively) for the enhancement of fingerprints in blood has been investigated. Optimal pH conditions and H2O2 concentrations have been determined using UV/Vis spectroscopy. Both OPD and PPD are effective and less hazardous alternatives to the presently used 3,3'-diaminobenzidine (DAB) for the development of blood fingerprints, especially on porous surfaces. The orange color of OPD and the purple color of PPD offer alternative colors to the brown color of DAB and the light green color of ABTS for standing out against particular backgrounds. Both OPD and PPD can be used after ninhydrin treatment, but the reverse is not the case.

3,3'-Diaminobenzidine↗

PubMed1979

[Notes on the transmitter histochemistry of the cortex cerebelli of rat].

The cortex cerebelli of the normal, adult rat was investigated fluorescence histochemically by means of the paraformaldehyde method according to FALCK and HILLARP (1962) for visualization of catecholamines and a modified ninhydrin-reaction for the demonstration of GABA (PFISTER and GORNE 1978), respectively. The Purkinje neurons were shown to display a considerable variability of the fluorescent GABA reaction product. Noradrenergic boutons were shown to be arranged mostly in a punctate manner around the somata of the Purkinje cells. The Purkinje cell pericarya would thus be under the inhibitory influence of cerebellar bascet neurons as well as of Locus coeruleus neurons.

Animals↗

PubMedDate not supplied

[Method of electrophoretic determination of S-methyl methionine in plants].

A method to determine S-methyl methionine (vitamin U) in plants has been devised. The method includes S-methyl methionine extraction extract purification on cationite KU-2 in the NH4+-form, S-methyl methionine eluation with 2n water solution of NH4OH from the cation exchange resin, eluate evaporation in vacuo, dissolution of the residue in a minimal amount of water acidified with hydrochloric acid, electrophoretic separation of S-methyl methionine from basic amino acids, ninhydrine development of the electrophoregram, methanol eluation of the spot corresponding to S-methyl methionine, and colorimetry of the solution at 506 mmu. Relative error is +/-2.5%.

Electrophoresis, Paper↗

PubMedDate not supplied

[Simultaneous estimation of catecholamines and serotonin after purification on ion exchange resin].

A general fluorimetric method was developed for determination of noradrenaline, adrenaline, dopamine and serotonin in a single sample. Purification of samples was carried out on columns of Dowex 50X4 and 10 in sodium form. As judget by an internal standard the yield of noradrenaline and adrenaline was about 90% and of dopamin -- about 75%. Serotonin was determined by means of a condensation reaction with ninhydrin. A more stable fluorescence of serotonin samples was provided by an increase in warming-up time up to 55 min. Treatment of control probes with iodine solution permitted to eliminate an unspecific fluoresence of the tissues studied. The yield of serotonin was about 75% as estimated by an internal standard.

Chromatography, Ion Exchange↗

PubMed2002

Interlaboratory variation of plasma total homocysteine measurements: results of three successive homocysteine proficiency testing surveys.

BACKGROUND: Numerous studies have demonstrated that increased plasma total homocysteine (tHcy), whether measured after fasting or after a methionine load, is associated with increased risk for cardiovascular and thromboembolic diseases. However, little information is available regarding interlaboratory variation of tHcy measurements, especially at the increased tHcy concentrations observed after loading. METHODS: We conducted three Homocysteine Proficiency Testing Surveys at 6-month intervals. Sets of five plasma pools with endogenous tHcy concentrations ranging from 5 to 48 micromol/L were sent to participants. We received 11, 23, and 17 responses in the first, second, and third surveys, respectively. The following methods were used by participating laboratories: fluorescence polarization immunoassay (FPIA); HPLC with fluorescent detection (HPLC-FD), further subdivided by type of reduction/derivatization; HPLC with electrochemical detection (HPLC-ED); amino acid analyzer with ninhydrin detection; and liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS). RESULTS: In surveys 1 and 2, no notable differences among the mean tHcy values obtained by the different methods performed were observed. In survey 3, tHcy values obtained by the FPIA method were significantly lower (P <0.05) at increased tHcy concentrations (34 micromol/L) compared with values obtained by HPLC-FD regardless of reduction/derivatization agents used. Our laboratory confirmed the observation that tHcy values obtained by the FPIA method differed from those obtained by HPLC-FD at increased tHcy concentrations by reanalyzing each pool 10 times by FPIA and HPLC-FD using tributylphosphine-ammonium 7-fluoro-2,1,3-benzoxadiazole-4-sulfonate (P <0.001 for tHcy >19 micro mol/L). The mean among-method variations in surveys 1, 2, and 3 were 19%, 12%, and 9.6%, respectively. When results of the three surveys were combined, the mean among-method variation on 170 samples was 13%. Within-method variation was lowest for the FPIA method (4.4%), and ranged from 11-20% for HPLC methods. CONCLUSIONS: Various degrees of imprecision and lack of correlation among tHcy methods indicate that there is a need to improve analytical precision, decrease analytical difference, and standardize tHcy measurements among laboratories.

Chromatography, High Pressure Liquid↗

PubMed1975

[Effect of the prolonged administration of chlortetracycline on the content of free amino acids and low-molecular peptides in the blood serum of rats].

Five individual ninhydrin positive fractions were found in the blood serum of rats in the control group with the help of high voltage electrophoresis in combination with preparative high voltage electrophoresis and paper chromatography. An analogous analysis of the blood serum of the test animals treated with 50 mg/kg of chlortetracycline for 12 days revealed 1 new fraction in the weak basic region. After the use of chlortetracycline for 18 days 2 new fractions in the strong acid region, 1 new fraction at the level of the electrophoretic mobility of asparaginic acid and 2 fractions in the region between neutral and basic amino acids were found. Analysis of the above fractions showed that the blood sera of the control and test rats contained in addition to the free amino acids peptides consisting of 3 to 9 amino acid residues. Some peptides isolated from the sera of the test animals contained amino acids, such as ornithine and citrulline which are not usually contained in proteins.

Administration, Oral↗

PubMed1975

A thin layer chromatographic method for the separation and identification of the tryptophan metabolites present in whole human urine.

This paper reports a new thin layer chromatographic system which is able to separate a total of thirtyeight (38) tryptophan metabolites from a synthetic mixture in a period of three hours. The system consists of thin layers of silica gel GF254 which are eluted in two dimensions, employing in the first dimension a mixture of propanone, 2-propanol, water, ammonia (0.88) (50 : 40 : 7 : 3 V/V) and in the second dimension a mixture of chloroform, glacial acetic acid, methanol and water (65 : 20 : 10 : 5 V/V). A method whereby the separated tryptophan metabolites may be unambiguously identified in situ following the thin layer chromatography is also presented and discussed. This method utilises the effect of ultraviolet light, at 254 and 350 nm, and the colour reactions between the metabolites and four reagents namely, a modified Ehrlich, o-phthaldehyde, Pauly and Ninhydrin-cadmium acetate. A summary of the unsuccessful attempts to quantitate the modified Ehrlich positive metabolites by in situ reflectance densitometry is also given. Finally, it is demonstrated that the new system may be used to advantage to separate and identify tryptophan metabolites in studies of whole human urine.

Child↗

PubMed1976

Studies on fluorescamine: Part I--Applications of fluorescamine in forensic toxicological analysis.

This paper describes some applications of the fluorescamine spot test to forensic toxicological analysis. The fluorescamine test only reacts with primary amines; thus, this test makes a clear-cut distinction between amphetamine and methamphetamine. Previous common spot tests used reacted the same with these two amines. Fluorescamine is 100 times more sensitive in detecting amphetamine extracted from urine on thin-layer chromatograms than ninhydrin. Thus, it is a more sensitive method of detecting amphetamine abuse in urinalysis screening programs.

Amphetamine↗

PubMed1999

[Purification and characterization of antifungal peptide LP-1].

An antifungal peptide LP-1 from Bacillus subtilis TG26 strain was purified by acid precipitation, acetone precipitation and Hi-pore reversed phase column chromatography. The molecular weight of LP-1 is 1057.3 D as determined by MALDI-TOF mass spectrometry, and its pI is 4.75 by PAG-IEF. It was also found to be thermostable. Its antifungal spectrum showed that LP-1 has strong inhibitory activity against many plant pathogenic fungi, such as Pythium aphanidermatum, Gibberella zeae, Alternaria longipe, Fusarium oxysporum f. lycopersici, etc.. The abnormal hyphal growth of Trichoderma viride caused by LP-1 such as swollen tips, twisted, short growth and cytoplasm condensation was also observed. Both ninhydrin reaction and peptide sequencing suggested that LP-1 is a cyclic peptide.

Anti-Bacterial Agents↗

PubMed2002

Pharmacological and haematological study of shol fish (Channa striatus) skin extract on experimental animal.

Snake head fish Channa striatus (locally called 'shol') skin extract (SFSE) was examined for certain pharmacological and haematological effects on experimental animals. LD50 of SFSE was found to be 6 mg/20gm (iv) in male albino mice. SFSE potentiated pentobarbitone induced sleeping time in male albino mice and produced hypothermia. Low dose of SFSE decreased respiratory rate in rat and guineapig and high dose produced apnoea leading to death. On isolated toad and guineapig heart, SFSE significantly decreased rate and amplitude of contraction leading to temporary blockade, which returned after repeated wash. On isolated nerve muscle preparations, SFSE produced irreversible blockade of twitch response. SFSE induced quick contraction on isolated guineapig ileum, which was antagonised by atropine and cyproheptadine. SFSE did not possess haemolytic and haemorrhagic activity but produced anaemia in male albino mice. A neurotoxic compound (fluoroscent and ninhydrin positive) was isolated from SFSE by thin layer chromatography. This compound (CS-NT) was lethal in male albino mice, produced death by apnoea in rat and produced irreversible blockade of isolated nerve-muscle preparation. This study confirms that the skin of Channa striatus possesses toxic, and lethal components, which needs further detailed study.

Anemia↗

PubMed2002

[Levels of amino group nitrogen and imidazole compounds in human oral fluid].

The procedures of definition amino nitrogen (sum amino acids) and imidazole compounds in human oral fluid are described. Amino nitrogen were determined by the ninhydrin method modified by the authors, imidazole compounds by the Pauli test. A total of over 20 patients with the relatively good dental status were surveyed. The level of amino nitrogen showed noticeable fluctuations (2.01 +/- 1.24 mmol per ml of oral fluid), whereas the content of imidazole derivatives was more stable (0.71 +/- 0.19 mmol/ml). Oral fluid samples displayed an in vitro high proteolytic activity, which explains contradictory results in the determination of the levels of amino nitrogen.

Amino Acids↗

PubMed1976

A screening method for protein characterization and differentiation.

A circular paper chromatographic method was developed for the separation of the amino acids in proteins into 7 subgroups. Butanol-acetic acid water (4+1+1) was used as the developing solvent. Eluted ninhydrin-stained aminograms gave rise to graphic profiles or numerical indexes based on absorbance percentages. The profiles can be used to compare protein-containing samples. Twenty different samples were studied through 190 comparisons of graphic profiles and coefficients of correlation, with only 4% misleading results. The method showed excellent reproducibility for the identification or differentiation of proteins and has the advantage of being performed with low-priced apparatus and reagents.

Amino Acids↗

PubMed2003

ESDA processing and latent fingerprint development: the humidity effect.

The influence of humidification in the ESDA process on subsequent development of fingerprints on paper items was studied. It was found that, while the DFO process is nearly insensitive to previous humidification, fingerprint development with ninhydrin or with indanedione can be significantly affected by previous humidification of the paper.

Journal Article↗

PubMed2003

The effects of latent print processing on questioned documents produced by office machine systems utilizing inkjet technology and toner.

Counterfeiting of currency and identity documents, death threats, illegitimate business transactions, and terrorist-related activities are some examples of the types of crimes that often involve documents produced from printers and copiers. Although standard protocol typically requires a questioned document (QD) examination prior to latent print (LP) processing, occasionally, items of evidence may be submitted for a QD examination following the application of a series chemicals utilized in the development of latent fingerprints. In such cases, the forensic examiner must take into account any previous treatments prior to initiating an examination on documents produced with a printer or copier. This study was devised to examine the effects of a latent print development technique [ninhydrin, physical developer, and a bleach enhancer] on the physical and chemical examination of documents produced from copiers and printers.

Journal Article↗

PubMed1975

[Histotopochemical investigations on elastic membranes of blood vessels with special regard to diabetes mellitus. I. Proteins (author's transl)].

Histochemical investigations on elastic membranes of vessels under normally and diabetic conditions have been accomplished. These studies were made on man (diabetic and non-diabetic subjects) and on rats with streptozotocin-diabetes. The results are comparable among one another. The amino acids histidine, tyrosine and tryptophan were not demonstrable. The detection of primary NH2-groups (ninhydrin-Schiff-method and o-diacetylbenzen-reaction) was positive however. The results of the reactions in healthy men and animals were more distinct than in diabetic human subjects and animals. In healthy children the intensities of the histochemically reactions were higher than in adults.

Adult↗

PubMed1992

[Photometric and chromatographic methods of measuring the concentration of acelysin in the air of a work area].

Two methods were applied to evaluation of acelysine level. Spectrophotometric one was based on reaction with p-nitrophenyldiasonium and evaluation of the product optic density at 510 nm. The second was chromatography in thin layer, where a mixture of n-butanol, formic acid and water was used as an eluent and 0.2% ninhydrin solution in butanol and 3 ml of ice acetic acid (60-70 degrees C) served as a developer. Both methods are supposed to evaluate acelysine level in the air of working area, when a half of MAC and admixtures are present.

Aerosols↗