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Four soybean meals (SBM) were prepared in a commercial solvent-extraction plant to give a much wider range in heat treatment than is usually found among commercially available SBM. The meals were designated in ascending order of heat treatment as Under, Normal, Over and Rumen Escape. The Normal meal was processed using standard operating conditions. The Under meal received less heat treatment by reducing the steam pressure and retention time in the desolventizer-toaster. Over and Rumen Escape meals received further heat treatment in an additional four-compartment toaster. The Over meal received less heat treatment than the Rumen Escape meal by reducing steam pressure and retention time in both toasters. Crude protein content was similar for the four meals, but lysine tended to decrease with increasing heat treatment. In general, urease activity, trypsin inhibitor, protein dispersibility index and nitrogen solubility index decreased with increasing heat treatment. The +a Hunterlab color values increased as heat treatment increased. Apparent ileal digestibility of N and amino acids were similar for all meals (P greater than .05); however lysine digestibility for the Rumen Escape meal was 3.3 percentage units lower than the average of the lesser-heated meals. Energy digestibilities and nitrogen balance data were also similar (P greater than .05) for the four meals, but the apparent biological value of the Rumen Escape meal was 4.5 percentage units lower than the average of the other meals. There were no differences in nutritional value among the Under, Normal and Over meals, which represent the range in heat treatment usually found among SBM. The Rumen Escape meal, which received more severe heat treatment, tended to have lower nutritional value than the lesser-heated meals.

Using brine shrimp lethality for activity-directed fractionation, goniothalamicin (I), a new tetrahydroxy-mono-tetrahydrofuran fatty acid gamma-lactone (acetogenin), has been isolated from ethanolic extracts of the stem bark of Goniothalamus giganteus Hook. f., Thomas (Annonaceae). This novel compound was found to be cytotoxic and insecticidal and inhibited the formation of crown gall tumors on potato discs. Annonacin (II), the only other reported mono-tetrahydrofuran acetogenin, was also isolated; the previously reported 9ASK (astrocytoma reversal) activity of II was confirmed, and II is now also found to be weakly active against 3PS murine leukemia.

A chemically defined medium (CC-494) was used to study the nutritional requirements of three spiroplasmas representing three distinct serogroups: flower spiroplasmas [Spiroplasma floricola and FS (SR-3)] and honeybee spiroplasma [HBS (AS-576)]. Glucose, fructose, and mannose were utilized by all three spiroplasmas. In addition, the honeybee spiroplasma could ferment trehalose, FS (SR-3) could ferment sucrose, and S. floricola could ferment trehalose, sucrose, and raffinose. The three spiroplasmas varied greatly in their requirements of amino acids for growth. S. floricola was the only strain that utilized arginine. HBS (AS-576) required at least one purine and one pyrimidine base (either free base or ribonucleoside) for growth, while both flower spiroplasmas grew with only one base in the medium. Oleic acid, cholesterol, and bovine serum albumin were essential to all three spiroplasmas. Palmitic acid, which was nonessential, promoted growth significantly.

The biosynthesis of ansatrienin (mycotrienin) has been studied in radioactive and stable isotope feeding experiments with Streptomyces collinus Tü 1892. The m-C7N unit of the ansa ring is efficiently and specifically derived from 3-amino-5-hydroxybenzoic acid; shikimic acid is not incorporated into this part of the molecule but does label the cyclohexanecarboyxlic acid moiety, providing all seven of its carbon atoms. Incorporation of methionine confirms origin of the methoxy group by transmethylation. The D-alanine moiety is derived directly from D-alanine rather than L-alanine. The terminal steps in the conversion of shikimic acid into cyclohexanecarboyxlic acid seem to be sequential reduction of 2,5-dihydrobenzoic acid and cyclohexene-1-carboxylic acid as evidenced by feeding experiments and the detection of a new ansatrienin containing a 1-cyclohexene instead of the cyclohexane moiety.

From two types of class V act mutants of Streptomyces coelicolor two monomeric precursors of actinorhodin have been isolated and their structures determined. One is the known antibiotic kalafungin and the other a new compound. Their relationship to actinorhodin biosynthesis is discussed.

A reverse-phase high-performance liquid chromatographic method has been developed to determine the sites of reaction and the product distribution of modified salmon sperm DNA. The DNA was reacted with methyl methanesulfonate in neutral solution, and then degraded into deoxyribonucleosides by snake venom phosphodiesterase and alkaline phosphatase. Four products were identified and quantitated: 7-methyldeoxyguanosine (37.1%), 7-methylguanine (7.3%), 3-methyldeoxycytidine (28.8%), and 1-methyldeoxyadenosine (26.8%). This method provides a rapid procedure for analysis of chemically or biochemically modified nucleic acids.

Pulmonary artery sling is an uncommon congenital vascular anomaly, which may cause significant respiratory morbidity resulting from an external tracheal compression during early infancy. This condition should be considered on finding a mass interposed between the trachea and the esophagus showing an anterior indentation on the esophagograms. The diagnosis is usually made by invasive methods such as bronchography, bronchoscopy, cardiac catheterization, and angiography. Magnetic resonance imaging (MRI) has now become an excellent method for diagnosing cardiovascular anomalies, but the use of MRI as a means of diagnosing pulmonary artery sling is rare. In 1988, Malmgren et al, first reported the use of MRI in three cases. Their findings suggested that two overlaping sets of axial sections might be necessary for depicting the anomaly because different parts of the left pulmonary artery are imaged in different slices. In this paper, we report a diagnostic scan image which utilizes a single axial scan and reveals the pulmonary artery sling arising aberrantly from the right pulmonary artery. We also found (a) when the pulmonary artery sling interposed between the esophagus and trachea; the trachea become stenotic below the upper one-third in LAO equivalent sagittal section; (b) the right pulmonary artery compresses the proximal part of right pulmonary bronchus, and the right upper lobe bronchus arises from the trachea independently in the coronal section. Thus, we conclude that MRI, if available, should be the first choice modality for further examination when a pulmonary artery sling is suggested by conventional radiography.

Quantitation of 7-methyldeoxyguanosine (m7dG) produced in the in vitro methyl methanesulfonate (MeMS) action on calf-thymus DNA is achieved by enzymatic degradation, liquid chromatographic separation and chemical ionization mass spectrometry. The total degree of methylation, measured by uptake of [14C]MeMS was 0.35%. Mass spectral analysis shows that m7dG constitutes 84% of the total methylated product. It is also shown that tandem mass spectrometry allows detection of m7dG, as the protonated base, down to 1 pmol level, suggesting that MS/MS analysis can be the method of choice in quantitation of the adducts of in vivo DNA modifications.

Piceatannol (3,4,3',5'-tetrahydroxy-trans-stilbene) (NSC 365798) has recently been isolated and was subsequently synthesized for NCI tumor panel testing as a new antileukemic natural product from the seeds of Euphorbia lagascae. During the synthesis, a bioactive reaction mixture of several partially O-methylated piceatannol analogues was obtained. This mixture has now been maximized and subjected to bioactivity-directed fractionation, using brine shrimp lethality, to yield 3,3',5'-tri-O-methylpiceatannol (NSC 381281); this new compound has improved stability and better bioactivity in several systems than piceatannol itself. To confirm the structure, 5 was synthesized from vanillin. In addition, the isovanillin analogue, 4,3',5'-tri-O-methylpiceatannol (NSC 381864), another new compound, was synthesized and found to be bioactive.

Glycolytic intermediates replaced glucose as carbohydrate sources in a chemically defined medium CC-494N, whereas keto acids of citric acid cycle intermediates and glyoxylate were supplemented with 10 mM glucose. Glucose-6-phosphate, fructose-1,6-bisphosphate, 3-phosphoglyceraldehyde, and pyruvate supported limited growth for Spiroplasma floricola (23-6), whereas only pyruvate supported SR 3 spiroplasma and S. melliferum (AS 576). alpha-Ketoglutarate at 5 mM or greater, hematin, 2,4-dinitrophenol, sodium fluoride, and arsenite were dramatically inhibitory, whereas acetic anhydride, tricarballylate, acetylene dicarboxylate, and sodium cyanide were mildly inhibitory at 10 mM or lower. Malonate at 50 mM was not inhibitory.

The 360 MHz 1H NMR spectra of the boron-containing macrolide antibiotic Aplasmomycin and of desboroaplasmomycin were analyzed to extract most of the parameters revealing the conformations of these compounds in CDCl3 solution. It was found than the conformation of aplasmomycin in CDCl3 solution is identical to that in the solid state and that removal of the boron atom from aplasmomycin results only in a slight conformational change of the molecule in CDCl3. All resonances observed in the 13C NMR spectrum of the antibiotic have been assigned on the basis of chemical shift theory, multiplicity analyses, single frequency proton decoupling experiments, comparison with several derivatives and model compounds, specific deuteration experiments, and analysis of one-bond carbon-carbon couplings of pairs of carbon atoms.

cDNAs corresponding to two types of mouse alpha 1-acid glycoproteins (AGPs), Agp-1 and Agp-2, were cloned and sequenced. Two alleles of Agp-1 were also identified from inbred mice by restriction site polymorphisms. One allele, Agp-1A, is associated with AKR/J, SJL/J, and SWR/J mice, while the other, Agp-1B, is found in BALB/c, DBA/2, C57BL/6, B10, and A/J mice. cDNAs of both Agp-1A and Agp-1B were cloned from BALB/c and (B10 x AKR/J)F1 mice. A single nucleotide substitution in the coding sequence of Agp-1 is responsible for the observed difference in electrophoretic mobilities of Agp-1A and Agp-1B. Several additional nucleotide variations in the 3' untranslated region were also found. In contrast to these two alleles, substantial nucleotide difference exist between Agp-1 and Agp-2. Agp-1 and Agp-2 contain five and six potential N-glycosylation sites, respectively.

Two new aristolactams, aristolactam-FI [1] and -FII [2], were isolated from active extracts of Pararistolochia flos-avis. Their structures were elucidated on the basis of nmr, ms, uv, and ir spectral data. Aristolactam-I [3] and -AII [4] were also isolated from this plant. Aristolactam-AII showed cytotoxicity against PS and KB cells in culture.

An investigation of Witheringia coccoloboides (Dammer) A. T. Hunziker [Capsicum fuscoviolaceum (Cufodontis) Mort. And Standl.] roots has resulted in the isolation of two cytotoxic compounds: physalin B (1) and a novel physalin, which was characterized by spectral analysis as 25,26-epidihydrophysalin C (2). Both compounds have demonstrated cytotoxic activity in 9KB and 9PS tumor cells (in vitro). Physalin B (1) has demonstrated moderate activity against the 3PS mouse leukemia (in vivo).

The risk and potential risk factors of pericardiocentesis were assessed by a review of a series of 52 pericardiocenteses comprising all those performed in the cardiac catheterization laboratory of one institution from 1971 to 1978. On the basis of the operative results, the patients were separated into two groups for comparison; Group I comprised all patients with a successful uncomplicated (35) pericardiocentesis and Group II all those with a nonproductive (16), nontherapeutic (1) or complicated (8) pericardiocentesis. Complications consisted of one death, one cardiac arrest, one aspiration of a subdiaphragmatic abscess and five ventricular punctures without adverse sequelae. Among the patients who had a nonproductive pericardiocentesis, the condition of 11 had probably been misdiagnosed but at least 4 had a false negative pericardiocentesis. Comparison of the two groups showed no significant difference in the incidence of cardiac tamponade or in the clinical presentation based on historical, physical, electrocardiographic, roentgenographic or echocardiographic findings of pericardial disease. Pericardiocentesis was usually successful when performed for suspected malignant pericardial effusion but often unsuccessful when performed for suspected hemopericardium. Anatomically, all patients in Group II had either minimal or loculated posterior pericardial effusion. It is concluded that pericardiocentesis can be performed at a low risk that can be further minimized by consideration of the disease process and the anatomic location of the pericardial fluid.

A highly sensitive (subpicomole level), structure-specific method of analysis has been developed for characterizing and quantitating the dinucleotide thymidylyl(3'----5')thymidine methyl phosphotriester by desorption chemical ionization tandem mass spectrometry. The methodology can be applied to related compounds such as the parent dinucleotide and employs either positive or negative ionization mass spectra or daughter spectra. The procedure allows specification of the site of methyl attachment.

A chemically defined medium, CC-494, supports the cultivation in vitro of several spiroplasmas belonging to three distinct serogroups. Medium CC-494 supports the growth of flower spiroplasmas FS 23-6 and FS SR-3 and of honey bee spiroplasma HBS AS-576. The maximal populations of the two flower spiroplasmas and honey bee spiroplasma are comparable to those cultured in the undefined medium C-3G. The growth rate for all three spiroplasmas is slower in the defined medium.

The antibiotic ketomycin is formed from shikimic acid via chorismic acid and prephenic acid. Phenylalanine and 2',5'-dihydrophenylalanine are not intermediates in the biosynthesis. Degradation of ketomycin derived from [1,6-14C]shikimic acid showed that prephenic acid is converted into ketomycin with stereospecific discrimination between the two enantiotopic edges of the ring, the pro-S-R edge giving rise to the C-2', C-3' side of the cyclohexene ring of ketomycin.

An NADP-linked, alcohol dehydrogenase from Entamoeba histolytica was purified to apparent homogeneity by Blue Sepharose affinity chromatography. Molecular weights of 130,000 and 30,000 were estimated by gel filtration and by sodium dodecyl sulfate polyacrylamide gel electrophoresis, respectively, suggesting that the enzyme is a tetramer. The enzyme exhibited more than 20-fold selectivity for NADP(H) over NAD(H). Although the purified enzyme acts on both primary and secondary alcohols, higher activity was found with secondary alcohols.

Screening of crude extracts of the bark of Annona bullata showed cytotoxic and pesticidal activities. By monitoring with brine-shrimp lethality, two novel, extremely potent acetogenins, bullatacin [1] and bullatacinone [2], were isolated. Spectral and chemical methods identified bullatacin as a diastereomer of asimicin. Bullatacinone represents bullatacin with the lactone cleaved and reformed at the 4-OH. Compounds 1 and 2 show selective cytotoxicities in human tumor cell lines, and certain susceptible cells give ED50 values as low as 10(-12)-10(-15) micrograms/ml. Bullatacin was pesticidal at concentrations as low as 1 ppm, but bullatacinone lacked pesticidal activities. The known compounds liriodenine and (-)-kaur-16-en-19-oic acid were also isolated and were lethal to brine shrimp but were not significantly cytotoxic.

Feeding experiments with 13C-labeled precursors followed by 13C NMR analysis of the antibiotic and its aglycon have established a polyketide mode of biosynthesis for chlorothricin, a metabolite of Streptomyces antibioticus. The acyl moiety is a substituted 6-methylsalicylic acid derived from four acetate units and a methyl group of methionine. The aglycon is comprised of ten acetate and two propionate units, leaving three carbon atoms, C-22, -23, and -24, unaccounted for. The two 2-deoxy-D-rhamnose moieties are derived from glucose with retention of the hydrogens at C-1, C-2, and C-6 and loss of H-3 and H-5. The hydrogen at C-4 of glucose is transferred intramolecularly to C-6 of the hexose, replacing the hydroxyl group at C-6 in an inversion mode, a result which implicates the thymidine 5'-diphosphate-glucose oxidoreductase reaction in this transformation.