Isolation and Identification of the Constituents of Volatiles of Dry Flowers of Aglaia odorata Lour

The headspace volatiles of the dry flowers of Aglaia odorata were trapped by a modified circulating adsorption method and were analyzed by GC/MS techniques after desorption by solvent. The components were identified by re, ass spectral and temperature-programmed retention index data on two types of glass capillary columns (OV-101 and PEG-20 M). 54 compounds have been identified from the volatiles, including 21 sesquiterpenes, 8 aldehydes, 7 alkyl benzenes, 6 alkanes, 6 other oxygen-containing compounds and I nitrogen containing compound. 29 compounds have not been reported in the studies on the constituents of the essential oil or absolute oil of the flowers.     

The Chemical Components of the Absolute Oil from Jasminun sarnbac (L.) Aiton

This plant is widely cultivated in southeastern and southern China. The concrete and absolute got from their flower are used in perfumery industry. It is also used popularly as ornamental plant and sometimes as medicine. The main chemical components of the absolute have been determined by the methods of chromatography, GC-MS, IR and NMR. The quantitative determination of these components was carried out by GLC also. The result is as follows: 3-hexenyl acetate 1.09%, cis-3-hcxen-l-ol 1.40%, dipentene oxide 1.09%, linalool 14.33%, methyl benzoate 2.96%, benzyl acetate 11.04%, benzyl alcohol 10.12%, cadinene 4.67%, hexenylbenzoat 16.51%, methyl anthranilate 0.62%, octadecene 6.54%, methyl oleate 2.02%, methyl' octadeca-9-ynoate 0.62%. Three minor components have been determined also.     

Studies on the Chemical Constituents of the Hydrocarbon Fraction of the Essential Oils from Caryopteris tangutica and C. Glutinosa

The chemical constituents of the hydrocarbon fraction of essential oils of Caryopteris tangutica Maxim. and C. glutinosa Rehd were examined by means of GC-MS-COM and GC retention time data on a 45 m capillary column coated with OV- 101. 21 components have been saparated and identified, i.e. α-thujene, α-pinene, cam- phene, sabinene, β-pinene, β-myrcene, α-terpinene, p-cymene, limonene, β-ocimene-y, β- phellandrene, α-terpinolene, α-cubebene, α-copaene, α-cedrene, β-caryophyllene, γ-cadinene, 1-aromadendrene, α-humulene, β-bisabolene and δ-cadinene. The quantitative determination of these components was carried out by GLC also.     

A new alloantigenic system associated with the Mls locus in the mouse.

An antiserum prepared by injecting C3H/HeJ mice with CBA/J tissue has been shown to react with cell surface components that are not part of any previously described system of serologically detectable alloantigens. The antiserum, which is designated AST-101, acts selectively in cytotoxic tests carried out with lymphoid cells, killing B cells, but not T cells. Phagocytic cells found in peritoneal exudates are also killed by AST-101 and complement in vitro; the sensitivity of other cell types has not been determined. Strain distribution does not indicate any association of the AST-101 system with H-2, Ly, or Thy systems; genetic analysis reveals close linkage with the mouse minor MLC-stimulating (Mls) locus. Serologic analysis also points to a close association between antigens reactive with AST-101 and the products of the Mls genes.     

Eel, Anguilla anguilla (L.), muscle modifications induced by long-term exposure to 101 ATA hydrostatic pressure

Fibre area, water and protein concentrations have been measured in red and white muscle of eels exposed for 1 month to hydrostatic pressure [HP = 101 atmosphere absolute (ATA)]. Comparison of the results with those from control fishes (1 ATA) shows that there is no variation of the measured parameters in red muscle. However, white muscle exhibits a significant decrease ( P < 0.01) in protein concentration with a concomitant decrease ( P < 0.001) in fibre area. It is proposed that long term exposure to HP = 101 ATA induces metabolic and/or morphological modifications which are dependent on muscle type.     

Isolation and Identification of the Constituents of Volatiles of Flowers of Chloranthus spieatus (Tunb.) Makino

The headspace volatiles of the flowers of Chloranthus spicatus (Tunb.) Makino were trapped by a modified circulation adsorption method and were analyzed by GC/MS techniques after desorption by solvent. The componends were identified by mass spectral and temperature-programmed retention index data on dual glass capillary columns (OV-101 and PEG-20 M). 32 compounds have been identified from the volatiles, including 11 monoterpenes, 11 sesquiterpenes, and 7 oxygen-containing compounds.The major consitituents are cis-methyl jasmonatecis-β-ocimene, and β-pinene.     

Hydrodynamic studies on interactions between the components of the liver microsomal cytochrome P-450 system

To understand the different behaviour of cytochrome P-450 systems in kinetics as well as in the demethylase activity, sedimentation and molecular weight experiments have been carried out with the following results: 1) Sedimentation coefficients of solubilized P-450 and P-450 LM2 fractions amount to 24 +/- 4 [S] and 12.8 +/- 1.2 [S], respectively. Molecular weights were determined to be 1.0 +/- 0.2 . 10(6) and 3.0 +/- 0.5 . 10(5) Dalton. 2) Triton N-101 provokes splitting of the associated structure both of solubilized P-450 and P-450 LM2; this effect is reversible. 3) The dissociation depends not only on the absolute concentration of Triton but rather on the Triton P-450 ratio. The dissociation curves of solubilized P-450 and P-450 LM2 are similar in shape and in the Triton/P-450 ratio dependence. 4) In the presence of small concentrations of Triton a more complicated dissociation behaviour was observed with broad integral distribution of the sedimentation coefficients. 5) The ionic detergent cholate splits the associated structure of P-450 LM2 at considerably higher concentrations in comparison with Triton-N 101. 6) Addition of reductase causes a decrease of sedimentation coefficients and molecular weights of solubilized P-450. The same effect in P-450 LM2 could be observed only in the presence of phospholipids.     

Rapid determination of ethylene glycol at toxic levels in serum and urine

The rapid gas chromatographic detection and determination of ethylene glycol in biological fluids is described. Phenylboronic acid in acetone was used for the esterification of glycol. The phenylboronates of ethylene glycol and 1,2-propylene glycol are not separated on a packed column of medium polarity (OV-17), but they can be separated on a non-polar column (OV-101). In both instances 1,3-propylene glycol can be used as an internal standard. The method requires only 100 μl of serum or urine and is suitable for trace analysis in an emergency toxicological laboratory. The utility of the method is demonstrated on two cases of human intoxication with ethylene glycol.     

A micromethod for the analysis of free amino acids by gas chromatography and its application to biological systems.

A gas chromatographic procedure was developed for determination of minute amounts of free amino acids in natural waters and laboratory models simulating biological systems. Sample pretreatment included removal of interfering organic substances by chloroform extraction and isolation of amino acids by cation exchange. Amino acids were converted to their N-heptafluorobutyryl isobutyl ester derivatives in glass capillary tubes, permitting considerable concentration of the sample prior to gc injection. The derivatives of 19 amino acids were successfully separated on either a glass column packed with a mixture of OV-101 and OV-17 on Chromosorb W, a glass capillary column coated with OV-101, or a support-coated capillary column supported with SE-30. One to five nanograms of individual amino acids were detected using flame ionization detector. The detection limit was reduced more than 100-fold using the electron capture detector and more than 1000-fold by mass fragmentography. The procedure allowed determination of less than 1 ppb of individual amino acids in lake and river water samples and was used to estimate the exeretion of free amino acids from microbial populations.     

The Fragrance Components of the Jasrninnm sambac (L.) Alton collected by Simultaneous Steam Distillation and Solvent Extraction

The volatile fragrance components of Jasminum sambac (L.) Aiton flower were collected by a modified simultaneous steam distillation and solvent extraction apparatus (SDE). The obtained extract was concentrated to 100 μL by the gas entrainment method in which the extract was distilled under reduced pressure with the influence of a steam of nitrogen. The concentrate was analyzed by GC-MS with PEG-20 M capillary column, and the Kovats indexes were determined at the same time whereby the nalkanes were added into extract. 28 compounds were identified. The major components were: linalool, benzyl acetate, ciscaryophyllene, cis-3-benzyl benzoate, methyl anthranilate and indole. These results show the components of the extract have no qualitative difference from absolute oil. The advantages of the above procedure were discussed in detail.     

Expression of mdm-2 oncoprotein in the primary and metastatic sites of mammary tumor (GI-101) implanted athymic nude mice

The expression of mdm-2 oncoprotein (p90) was determined in a human breast tumor xenograft line (GI-101) that was derived from a 57 year old female cancer patient with recurrent, infiltrating ductal adenocarcinoma (Stage IIIa, T3N2MX). Immunoprecipitation coupled western blot analysis of the primary tumors that have been obtained from xenograft implanted athymic nude mice, using mdm-2 (Ab-1) mouse monoclonal antibody, primarily revealed high level expression of a 90 kD full length mdm-2 protein. In the GI-101 tumor the level of full length mdm-2 (p90) protein expression increased with the increase in the size of the tumor (100 to 2,000 mm(3)) and a maximum expression was detected in 2,000 mm(3) size tumors. In addition to the expression in the primary site, a significantly high level expression of mdm-2 protein (p90) was detected in the lung and liver tissues also, which are the known metastatic sites for GI-101 xenograft tumors. However, the level of mdm-2 protein expression was undetectable in the lung and liver tissues obtained from control mice. A cell line (GI-101A) derived from the GI-101 xenograft tumor also showed a high level expression of mdm-2 protein after several generations of cell passage. When the GI-101A cells were treated with DES (Diethylstilbestrol) the mdm-2 protein expression increased after 10 min treatment and reached a peak level at 40 min. Interestingly, DES (10 and 20 microM) treatment increased the total cell number also after 96 hr treatment compared to the non-treated cells. It appears that mdm-2 (p90) may have a significant role in supporting the tumor cell growth as well as the metastatic process of the GI-101A cells.     

The Main Chemical Components of the Essential oil from Rosa rugosa Thunb

The plant of Rosa rugosa Thunb. now is cultivated widely in northern China and other places is characterized as Rosa rugosa cv 'Plena' with violet-redflowers, The essential oil extracted by steam distillation, from its flower is a famousfragrance: The flower can be used as medicine and food. The chemical components ofthis essential oil have been determined by chromatography, IR, DC-MS and other methods. Thirty components have been identified, such as β-citronellol, geraniol, geraniolacetate, citronellyl acetate, methyl eugenol, linalool etc. Their contents in the oil havebeen determined also. The chemical components of absolute oil have been determined byGC-MS. All these results can be used to compare with that of Rosa damascena Millfrom Bulgarian 'Valley cf rose and other Rosa sp.     

Aspects of the reproductive biology of the bass, Dicentrarchus labrax L. II. Fecundity and pattern of oocyte development

The European bass, Dicentrarchus labrax L., in common with most temperate marine teleosts, exhibits a distinct seasonal reproductive cycle. Its reproductive strategy is characterized by a high absolute fecundity (2-25 × 10⁵), both absolute and relative fecundity increasing with size and age. The cyclic pattern of oocyte development has been determined by histology and from oocyte size frequency analysis. Results show that the bass exhibits group-synchronous oocyte development and is a 'fractional spawner', i.e. it spawns a number (3–4) of discrete clutches in quick succession, successive clutches containing fewer oocytes. All oocytes which are recruited into the secondary growth phase (> 110 μrn) are either spawned that season or become atretic, i.e. no secondary oocytes are held over for the subsequent reproductive season.     

Pentacyclic triterpenoids and sterols from seven species of mangrove

The isolation of pentacyclic triterpenoids from seven species of fresh mangrove leaves using a simple and rapid method is described. The leaves were homogenized using chloroform—methanol and the extract was diluted with water to precipitate out triterpenoids which were separated into neutral and acidic fractions. These were analysed by gas-liquid chromatography as acetyl and trimethylsilyl ether derivatives on a 3% OV-17 column. Sterols were isolated from the chloroform layer by preparative thin layer chromatography and were analysed by gas-liquid chromatography as their trimethylsilyl ether derivatives on a 3% OV-17 column. The triterpenoids found were α-amyrin, β-amyrin, lupeol, oleanolic acid and ursolic acid in most of the samples. Sterols found in all the samples were cholesterol, campesterol, stigmasterol, sitosterol and stigmast-7-en-3β-ol. Retention indices of the triterpenoids and sterols have been determined.     

Morphometric and stereological study of the seminal vesicle of the guinea pig.

The seminal vesicle of the guinea pig has been widely used as a model for the study of hormonal action on the male accessory sex organ, but there have been few attempts to quantify their cellular and tissue components. In the present study, the seminal vesicle of the guinea pig was described in the form of a morphometric model. Tissue samples were taken from the distal, middle and proximal regions of the gland and processed for light microscopy. Using a combination of a stereological point-counting technique and direct measurement, the relative volumes of different components (lumen, epithelium, lamina propria and fibromuscular layer) were determined. The relative numbers of the secretory cells and basal cells were also estimated. Following the estimation of the average size of the seminal vesicle, the relative volume of different components and the relative number of secretory cells were transformed into absolute data on a per average seminal vesicle basis. Similarly, the average sizes of the secretory cells and nuclei were also determined. The quantitative data generated from the present study will serve as a baseline for further studies of the seminal vesicle of the guinea pig. The techniques used in the present study are easy to apply, and data generated were objective and reproducible.     

Method for analysis of polybrominated biphenyls by gas chromatography mass spectrometry

Gas chromatography using a short packed column (45 cm, 0.2 cm i.d., 2% OV-101 on Gas-Chrom Q) with mass spectrometric detection in the selected ion monitoring mode has been found satisfactory for the analysis of lower as well as higher polybrominated biphenyls. Acceptable sensitivity (< 1 ng) may be achieved for this method by focusing selectively at either the low (m/z 20-600) or the high m/z 600-1000) range of the quadrupole filter (low range for mono- through hexabromobiphenyl, high range for hexa- through decabromobiphenyl). A tuning technique has been developed for low range and high range polybrominated biphenyls using the ion abundances of perfluorotributylamine as a standard. Standard ions for the quantitation of mono- through decabromo-biphenyls were selected and validated. The technique was applied to the analysis of a variety of environmental samples.     

Multiple protein binding sites within the ovalbumin gene 5''-flanking region: isolation and characterization of sequence-specific binding proteins.

To understand the molecular basis of the steroid hormone regulated expression of the ovalbumin gene, we sought to identify and isolate nuclear factors from chicken oviduct which interact specifically with the ovalbumin promoter. Using DNase I footprinting and mobility shift assays, we have defined at least four distinct protein binding sites, OV-150, OV-220, OV-250 and OV-330, in the promoter region between -100 to -400. Binding competition and protein fractionation studies revealed the existence of two distinct proteins, each recognizing two promoter sites: Both OV-330 and OV-250 are recognized by one protein factor which is distinct from the one binding to both OV-220 and OV-150. The location of the DNase I footprints coincides with those of in vivo chromatin hypersensitive sites. The OV-330 site is located in a sequence area required for the repression of the gene in the absence of hormone. The factor binding to OV-330 has been substantially purified and renaturation experiments indicate that the binding activity is associated with a polypeptide(s) of Mr 40K.     

Automatic collection of samples from suspension cell cultures for investigations of prolonged metabolic processes

A glc method has been described for the quantitative analysis of tobacco (and other plant tissue) for esculetin (as free esculetin and the glucosides esculin or cichoriin) which used tritium-foil electron-capture detection of as little as 15 ng esculetin. Polyphenols were directly extracted from plant material, using n-propanol, and the esculetin glucoside was hydrolyzed with n-propanol saturated with HCl gas to produce free esculetin. After silylation, the TMS-esculetin was separated on 10% OV-101, with caffeic acid n-butyl ester as the internal standard.     

Immunologic effects of AS101 in the treatment of cancer patients

AS101 [ammonium trichloro(O,O'-dioxyethylene)tellurate] is a new immunomodulator previously shown to stimulate the production of different cytokines in vitro and in vivo. We report here our results of a phase I clinical trial conducted on 47 cancer patients with advanced malignancies. AS101 was administered intravenously at escalating doses from 1 to 10 mg/m2, twice or thrice a week. The maximal tolerated dose has not yet been determined. However, significant immunologic responses were noted at dose levels of 1-3 mg/m2 administered three times a week. At these doses statistically significant rises in gamma-interferon, natural killer cell activity, tumor necrosis factor and interleukin-2 (IL-2) levels as well as the expression of IL-2 receptors were noted. In most of the immunologic parameters the maximal response was seen at 3 mg/m2. Throughout the study toxicity was minimal. In view of these results phase II studies are currently being initiated.     

Sensitive detection of 5-methylcytosine and quantitation of the 5-methylcytosine/cytosine ratio in DNA by gas chromatography--mass spectrometry using multiple specific ion monitoring.

A method combining gas chromatography and mass spectrometry (GC-MS) with multiple specific ion monitoring has been developed for the detection of 5-methylcytosine and the quantitation of the ratio of methyleytosine to cytosine in DNA. The trimethylsilyl derivatives of cytosine and 5-methylcytosine obtained from DNA hydrolysates are separated by isothermal elution on an OV-225 column and detected by specific ion monitoring in a DuPont 321 mass spectrometer. As little as 1.6 pmol of 5-methylcytosine in Φ_χ174 DNA can be detected, corresponding to a tenfold improvement in sensitivity over that obtained by conventional techniques. The ratio of 5-methylcytosine to cytosine of DNA from φ_χ174, calf thymus, salmon sperm, and several mouse tissues has also been determined. The results agree well with those obtained by other methods.