Lack of biologically active estrogens in commercial cow milk.

Recently, many studies have investigated potential estrogenic compounds in the human diet. Several of these investigations have studied cow milk, a mainstay of the diets of both young and old. In vitro studies have determined that estrogens can be found in milk, and that the concentration of estrogen may be correlated to the fat content in the milk. Regardless, the majority of these studies have concluded that the levels of estrogens in milk are too low to have a physiological effect. However, a recent study found that commercial 1% cow milk was uterotrophic in rats, suggesting that it contained biologically significant levels of estrogen. Using the rat model, we tested milk samples from commercial sources and with varying fat content for estrogenic activity. Ovariectomized female rats were given milk ad libitum for a period of 2 wk. After 12 d of treatment, rats were tested sequentially in an open field and an elevated plus maze to determine any effect of milk on anxiety levels. Upon completion of the behavior testing, uterine weights were examined. Regardless of milk type, no difference was observed in daily volume of milk consumed. Contrary to previous publications, no differences existed in either the behavior or the uterine weights between animals that consumed any milk type and the negative controls. These results demonstrated that none of the commercial milk types that we tested contained biologically significant estrogenic activity.

Increased behavioral neurosteroid sensitivity in a rat line selectively bred for high alcohol sensitivity.

Acute administration of a neurosteroid 5beta-pregnan-3alpha-ol-20-one induced a greater impairment in motor performance of the selectively bred alcohol-sensitive (ANT) than alcohol-insensitive (AT) rats. This difference was not associated with the sensitivity of gamma-aminobutyrate type A (GABA(A)) receptors, as 5alpha-pregnan-3alpha-ol-20-one (allopregnanolone) decreased the autoradiographic signals of t-butylbicyclophosphoro[35S]thionate binding to GABA(A) receptor-associated ionophores more in the brain sections of AT than ANT rats. Nor was the difference associated with baseline levels of neuroactive progesterone metabolites, as 5alpha-pregnan-3,20-dione (5alpha-DHP) and 5alpha-pregnan-3alpha-ol-20-one were lower in the ANT rats. After ethanol (2 g/kg, i.p.) administration and the subsequent motor performance test, the increased brain concentrations of these metabolites were still lower in the ANT than AT rats, although especially in the cerebellum the relative increases were greater in the ANT than AT rats. The present data suggest that the mechanisms mediating neurosteroid-induced motor impairment are susceptible to genetic variation in rat lines selected for differences in ethanol intoxication.

[Determination of 4-bromo-2,5-dimethoxyamphetamine (DOB) found in illicit tablets seized in Italy]. / Identificazione della 4-bromo-2,5-dimetossiamfetamina (DOB) in compresse clandestine sequestrate in Italia.

Some of the molecules belonging to the amphetamines group (4-bromo-2,5-dimethoxyamphetamine, DOB) or to the phenethylamines (4-bromo-2,5-dimethoxy-phenethylamine, 2C-B or Nexus) have closely related structures that make their identification quite difficult. The unambiguous identification is crucial in forensic responses. This paper describes the analytical approach used to achieve the identification of the main ingredient contained in tablets seized in the illicit market of Rome (Italy) and submitted to our laboratory by the Court of Law of Rome. The procedure entails the basic extraction of the main ingredient from the tablets with tert-butyl methyl ether followed by qualitative gas chromatographic mass-spectrometric (GC-MS) analysis using both electron impact detection (EI) and chemical ionization (CI). The examination of the mass spectra obtained from the native molecule and from its pentafluoropropionyl-derivative allows the structural identification of the side chain and the substitutions on the aromatic ring. This analytical approach can thus be useful to distinguish between amphetamine-like and phenetylamine-like compounds using instruments and techniques commonly available in the forensic toxicology laboratories.

Identification of 3,4-methylenedioxyamphetamine analogs encountered in clandestine tablets.

Recently, 3,4-methylenedioxyamphetamine derivatives have been encountered in the Italian illicit market, mainly in form of tablets. Among this class of substances small modifications of the molecule may result in a wide range of derivatives and analogs some of which are not yet listed as controlled substances in the Italian schedules. Due to the structural similarity some of these molecules have a gas chromatographic behavior and mass spectra that only slightly differ. In the present work, an analytical strategy is proposed to achieve the identification of analogs within this class of molecules. In seized material sent by the Court of Law of Rome to our laboratories a number of tablets engraved with different symbols (e.g., 'Dollar', 'Fido Dido' and 'Bomb') were submitted to analysis in order to establish whether they contained drugs of abuse. The analytical techniques employed for this purpose were UV spectrophotometry and thin-layer chromatography which provided information suggesting that the tablets contained a methylenedioxyamphetamine. Gas chromatography with flame ionization detection indicated that the main ingredient differed from the molecules of the same class already known. Finally, capillary gas chromatographic-mass spectrometric analysis of the native molecules and their pentafluoropropionic acid derivatives, performed with both, electron impact and chemical ionization, allowed the identification, in each tablet, of three molecules: the N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MDP-2-MB, MBDB), the 1-(3,4-methylenedioxyphenyl)-2-butanamine (MDP-2-B) and the N,N-dimethyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MDP-2-MMB).

Marked decrease of plasma neuroactive steroids during alcohol withdrawal.

The neuroactive steroids allopregnanolone (ALLO) and allotet-rahydrodeoxycorticosterone (THDOC) are the most potent endogens positive modulators of gamma-aminobutyric acid (GABA) on GABAA receptors, a receptor system presumably responsible for some behavioral responses to alcohol withdrawal. In a group of nine alcoholic subjects, the levels of plasma ALLO and THDOC were markedly lower than those of control subjects during the early withdrawal phase (day 4 and 5), when anxiety and depression scores were higher. In contrast ALLO and THDOC plasma levels did not differ from those of control subjects during the late withdrawal phase when anxiety and depression scores were low. These results suggest that the decrease of neuroactive steroid biosynthesis may contribute to the withdrawal symptoms.

Comparative analysis of illicit heroin samples.

The comparative analysis of street heroin samples is still an object of scientific discussion. A combination of a wide range of analytical techniques is necessary to obtain a valid amount of information about the sample composition, and it is impossible to define an unique analytical approach. We suggest a complete analytical sequence based on analysis of volatile compounds, opiates, diluents, adulterants and metals, by head space gas chromatography (HS/GC), gas chromatography mass spectrometry (GC/MS), thin layer chromatography (TLC), high pressure liquid chromatography (HPLC) and atomic absorption (AA) using a sample amount as low as 50-100 mg. The outlined procedure can be successfully applied to routine work, thus obtaining suitable information about a sample's chemical composition. This helps to attribute or exclude common sources of separate specimens. Results obtained on 33 street heroin samples confiscated in the metropolitan area of Rome are listed.