[Bouveret's syndrome. Two new cases (author's transl)]. Pyloro-duodenal obstruction by a gall-stone which has passed through a bilio-digestive fistula (Bouveret's syndrome) is rare. The authors report two cases. The clinical picture is that of pyloric obstruction. The diagnosis, suspected on barium meal, is today confirmed by fibre endoscopy. The treatment is fairly simple: cholecystectomy, removal of the gall stone, closure of the fistula; the prognosis depends on the general context.


[Lumbar hernia (author's transl)].

[Lumbar hernia (author's transl)]. A case of lumbar hernia is reported here. The likelihood of the traumatic etiology of this case and the technics of parietal reparation are discussed. The risk of strangulation justify the preventive treatment.


[Histological study of livers conserved for long periods (author's transl)]. 33 pig livers were preserved using two comparative methods: extra-corporeal perfusion and immersion in hypothermia. The duration of conservation varied between 2 and 24 hours. A macroscopic and microscopic study carried out in all cases permitted us to define four cytological stages for the lesions, completed by an ultrastructural study showing the cyto-pathological course of the hepatocytes in two distinct cell series. The interest of this study is to better appreciate the degree of aggression of the lobule in relation to the metabolic gradient and that of the hepatic parenchyma. Analysis of the results is in favour of conservation of the liver by immersion.


[Reaction of some suture materials on the process of intestinal healing. Experimental study (author's transl)]. The authors studied the influence of some suture materials (normal and chromic catgut, silk, polygalactic acid and polypropylene) on the process of healing of the small intestine in the rabbit. From the macroscopic, microscopic and mechanical examinations, it results that polygalactic acid is the material which comes closest to the ideal material. There follow, in that order: silk, polypropylene, chromic catgut and normal catgut.


Capillary column gas chromatographic profile analysis of volatile compounds in sera of normal and virus-infected patients. Using a transevaporator sampling technique, the volatile profiles from 70 microliter of serum were obtained by capillary-column gas chromatography. The complex chromatograms were interpreted by a combination of manual and computer techniques and a two-peak ratio method devised for the classification of normal and virus-infected sera. Using the K-nearest neighbor approach 85.7% of the unknown samples were classified correctly. Some preliminary results indicate the possible use of the method for the assessment of virus susceptibility.


Urinary phenylethylamine excretion: gas chromatographic assay with electron-capture detection of the pentafluorobenzoyl derivative. Phenylethylamine was extracted into n-hexane from alkaline urine saturated with sodium chloride, and back-extracted into dilute acid. The extract was freeze-dried and the residue converted to a pentafluorobenzoyl derivative for analysis by gas chromatography on a column of OV-225 with electron-capture detection. Quantification was achieved by adding an internal standard of tolylethylamine to each sample prior to extraction. Output values in normal subjects and in some patients with phenylketonuria and hyperphenylalaninaemia were in agreement with those in some other recent reports.


Determination of plasma and urinary cortisol by high-performance liquid chromatography using fluorescence derivatization with dansyl hydrazine. A method is described for the determination of cortisol in human plasma and urine by high-performance liquid chromatography using fluorophotometric detection. After extraction with methylene chloride, cortisol is labelled with dansyl hydrazine, and then separated by high-performance chromatography. The eluate is monitored by a fluorophotometer at 350 nm (excitation) and 505 nm (emission). The optimum conditions for the determination, such as HCl and dansyl hydrazine concentrations, reaction time and reaction temperature, and for the eluent of high-performance liquid chromatography, are discussed. Linearity of the fluorescence intensity (peak height) with the amount of cortisol was obtained between 0.5 and 60 ng. The recoveries for 50 and 100 ng of added cortisol were 98.7 and 95.4% for plasma, and 96.4 and 90.6% for urine, respectively. Comparison with a radioimmunoassay gave a correlation coefficient of 0.978. The proposed method is suitable for the routine analysis of cortisol in plasma and urine.


High-performance liquid chromatographic determination of free and total polyamines in human serum as fluorescamine derivatives. A highly sensitive and simple fluorimetric method for the determination of free and total polyamines, spermidine, spermine, putrescine and cadaverine, in human serum by high-performance liquid chromatography is described. The polyamines, obtained after clean-up of deproteinized serum by Cellex P column chromatography, are converted to their fluorescamine derivatives in the presence of nickel ion which inhibits the reaction of interfering amines with fluorescamine, and the derivatives are separated simultaneously by reversed-phase chromatography (LiChrosorb RP-18) with a linear gradient elution. The lower limits of detection are 10 and 15 pmole for spermine and the others in 0.5 ml of serum, respectively.


Measurement of mephenytoin (3-methyl-5-ethyl-5-phenylhydantoin) and its demethylated metabolite by selective ion monitoring. Mephenytoin (3-methyl-5-ethyl-5-phenylhydantoin) and its demethylated metabolite Nirvanol (5-ethyl-5-phenylhydantoin) were measured by a selective ion monitoring technique. This method was used in the analysis of both compounds in plasma from a patient receiving 50 mg and 400 mg of mephenytoin in single oral doses. Both compounds were extracted from plasma and ethylated prior to analysis by electron-impact mass spectrometry. Alkylation, using ethyl iodide in 2-butanone, occurred in the N-1 and N-3 positions of the hydantoin ring when concentrated KOH was added to the reaction mixture. The lower limits of quantitation for mephenytoin and Nirvanol were 10 ng/ml and 50 ng/ml, respectively, and were found to be reproducible within 8% upon repeated quantification.


Isolation by high-performance liquid chromatography and quantitation by radioimmunoassay of therapeutic concentrations of digoxin and metabolites. A method combining high-performance liquid chromatography for separation of digoxin from three of its metabolites with subsequent quantitation of each compound by radioimmunoassay is described. The metabolites are shown to interfere with the radioimmunoassay procedure thus providing the need for separation prior to assay.


Determination of plasma concentrations of dapsone, monoacetyl dapsone and pyrimethamine in human subjects dosed with maloprim. A high-performance liquid chromatographic method was developed to enable dapsone, monoacetyl dapsone and pyrimethamine to be measured simultaneously in plasma samples from volunteers in England and Malaysia who had been dosed with Maloprim. Mean half-lives of 25 and 80 h were calculated for dapsone and pyrimethamine, respectively, but there was wide individual variation. All subjects were found to be classifiable as "slow acetylators".


Simultaneous determination of the anticonvulsants, cinromide (3-bromo-n-ethylcinnamamide), 3-bromocinnamamide, and carbamazepine in plasma by high-performance liquid chromatography. A high-performance liquid chromatographic method is described for monitoring plasma concentrations of cinromide (3-bromo-N-ethylcinnamamide) and its de-ethylated metabolite. Carbamazepine levels can be easily measured by the same technique. The N-isopropyl analogue of cinromide is used as internal standard, and all compounds are easily separated on a reversed-phase column operated at 55 degrees with a small-diameter pre-column maintained at the same temperature. The extraction is rapid and generally applicable to plasma and urine samples that are to be analyzed by reversed-phase chromatography. Short- and long-term reproducibility studies show less than 4% relative standard deviation for replicate determinations for all drugs. Limits of quantitation are 10-20 ng/ml with an internal standard concentration of 3 micrograms/ml. Another metabolite of cinromide, 3-bromocinnamic acid, which may have some anticonvulsant effect, can be analyzed simultaneously by buffering the mobile phase and adding an ion-pairing reagent.


Quantitative thin-layer chromatographic measurement of n-trifluoroacetyladriamycin-14-valerate (AD 32) and trifluoroacetyladriamycin (AD 41) in blood and tissues. A thin-layer chromatographic method has been developed for the detection and measurement of N-trifluoroacetyladriamycin-14-valerate (AD 32) and its major metabolite trifluoroacetyladriamycin (AD 41). The procedure gives satisfactory linearity over a large range of concentrations. The coefficient of variability is about 10% over the entire range of usable concentrations, giving good reproducibility; sensitivity is 25 ng for both AD 32 and AD 41. Analysis is specific for AD 32 and AD 41 since adriamycin or more polar metabolites can be differentiated. Recovery is high (85-90%) and the method is simple and economical to use. Pharmacokinetics of AD 32 and AD 41 are reported in blood and some tissues of mice bearing Lewis Lung carcinoma.


Determination of sulfinpyrazone and two of its metabolites in human plasma and urine by gas chromatography and selective detection. A selective and sensitive gas chromatographic method for simultaneous determination of sulfinpyrazone and two of its metabolites (the para-hydroxylated metabolite and the sulfone metabolite) in biological fluids using alkali flame ionization detection (AFID), electron capture detection (ECD) and mass fragmentographic detection is described. The compounds are extracted from the samples, methylated and separated on 2% OV-17 or 3% OV-225 columns. Phenylbutazone is used as internal standard. Standard curves are linear. The coefficient of variation at 10 microgram/ml of sulfinpyrazone in plasma was shown to be 1.8% (AFID), and the detection limits were 0.1 microgram/ml (AFID) and 10 ng/ml (ECD). Mass spectra of the methylated compounds are shown and serum concentration curves after oral administration of 100 mg sulfinpyrazone to two persons are determined together with the excreted amounts of drug and metabolites.


Simultaneous determination of griseofulvin and 6-desmethylgriseofulvin in plasma by electron-capture gas chromatography. Two methods have been developed for the simultaneous determination of griseofulvin and its major metabolite 6-desmethylgriseofulvin in plasma using electron-capture gas chromatography. The first method was based on the quantitative reversion of the 6-desmethyl metabolite to griseofulvin by diazomethane. Plasma extract was chromatographed both before and after treatment with diazomethane, the former being the measure of griseofulvin and the latter representing the sum of the two compounds. In the second method, plasma extract was treated with diazobutane and griseofulvin and the butylated metabolite were separated by gas chromatography. The sensitivity for griseofulvin was 20 ng/ml by both methods and that for the metabolite was 20 ng/ml and 40 ng/ml by the first and the second method, respectively. The concentrations of the metabolite as well as griseofulvin were determined in dog and human plasma after oral administration of griseofulvin.


Profiling of uremic serum by high-resolution gas chromatography-electron-impact, chemical ionization mass spectrometry. A fast and reliable procedure for gas chromatographic profiling of components in ultrafiltrated uremic serum has been developed, using glass capillary columns. Sample pretreatment consists of ultrafiltration, evaporation and silylation. Some twenty components are identified by electron-impact and chemical ionization mass spectrometry. A comparison is made between profiles of sera from a series of uremic patients, before and after hemodialysis, and from non-uremic sera. Significant differences are found between these profiles. A "dialysis ratio" is introduced as a parameter for the removal of retained components by hemodialysis treatment.


Comparison of different high-performance liquid chromatographic systems for the purification of adrenal and gonadal steroids prior to immunoassay. The high-performance liquid chromatography of nineteen hormonal steroids with special respect to its suitability for routine purification of these steroids from crude, organic extracts of biological fluids prior to final quantitation by immunoassay has been studied. In all systems the gradient elution technique was applied. Separation of steroids has been investigated using different stationary phases chemically coated with non-polar, hydroxyl, NO2 and CN groups. Reproducibility of retention times was studied on a stationary phase coated with hydroxyl groups (DIOL column) using different organic eluents. Coefficients of variation range from 0.76 to 8.16%. Reproducibility was shown to be unequivocally better in the gradient part than in the isocratic part of the chromatographic run. In contrast to the other steroids, 18-hydroxylated steroids were more or less unstable in certain systems studied. As to resolution and reproducibility, the DIOL column run with an n-hexane-dioxane gradient has been shown to be superior to the other systems studied.


Determination of cortisol in human plasma by reversed-phase high-performance liquid chromatography. A method is described for the measurement of cortisol in human plasma using 45% aqueous methanol eluent on a 120 mm x 4.5 mm I.D. Hypersil octadecylsilane column with UV detection at 239 nm after a simple dichloromethane extraction and evaporation with a prednisone internal standard. The sample preparation time and chromatography time are each about 15 min and linear correlations have been obtained with plasma samples assayed by the Mattingly fluorimetric technique and a commercial-kit competitive protein binding method. Concentration down to 30 nmol/l may be measured and the method can be used when fluorimetry is invalidated by interference, particularly from spironolactone.


Determination of 5-dimethylaminonaphthalene-1-sulfonyl derivatives of urinary polyamines by ion-pair high-performance liquid chromatography. A sensitive and specific method for the determination of diamines and polyamines by ion-pair high-performance liquid chromatography is described. The 5-dimethylaminonaphthalene-1-sulfonyl derivatives of putrescine, 1,6-diaminohexane, spermidine and spermine are separated on a muBondapak C18 reversed-phase column with 1-heptanesulfonic acid and acetonitrile as the mobile phase. All compounds are eluted within 30 min using a programmed solvent gradient system. The method has a lower detection limit of 1 pmole on column. Because of the simplicity of the method, its application provides a better means for closely monitoring patients undergoing treatment for various types of genito-urinary neoplastic diseases.


Determination of dopa, dopamine, dopac, epinephrine, norepinephrine, alpha-monofluoromethyldopa and alpha-difluoromethyldopa in various tissues of mice and rats using reversed-phase ion-pair liquid chromatography with electrochemical detection. A method for the determination of catecholic amino acids and amines by reversed-phase ion-pair high-performance liquid chromatography with electrochemical detection has been developed. B using octanesulfonic acid for ion pairing and by optimising ionic strength, pH and methanol concentration of the mobile phase, separation was achieved of 3,4-dihydroxyphenylalanine (DOPA), 3,4-dihydroxyphenylacetic acid (DOPAC), norepinephrine (NE), epinephrine (EPI), and dopamine (DA). Alpha-Difluoromethyldopa (DFMD) and alpha-monofluoromethyldopa (MFMD), two potent enzyme-activated irreversible inhibitors of aromatic amino acid decarboxylase were also separated from the natural catechols. Concentrations of catechols and inhibitors were measured in brains, hearts and kidneys of mice treated with small repeated doses of MFMD. The method has also been applied to the determination of catechols in other organs such as prostates and seminal vesicles of rats and in smaller tissues like mesenteric arteries. A semi-automated procedure making use of an automatic sample processor and a digital integrator permitted the analysis of as many as sixty samples per day.


Quantitative analysis of volatile halothane metabolites in biological tissues by gas chromatography. A simple and sensitive gas chromatographic method for the determination of 2-chloro-1, 1-difluoroethylene (CDE) and 2-chloro-1,1,1-trifluoroethane (CTE), two highly volatile metabolites of halothane, in blood, liver and isolated hepatic microsomes is described. The entire head-space in equilibrium with a known volume or weight of the sample is injected into the gas chromatograph equipped with a flame ionization detector. Quantification is accomplished with standards prepared by fortifying blank samples with known concentrations of CDE and CTE which are treated under the same conditions as the samples. Detection limits for CDE and CTE were 2 pmole/ml in blood and 10 pmole/g in liver and the mean relative standard deviations are no greater than +/- 6% except for CTE in hepatic microsomes (+/- 9%). A preliminary study of blood CDE and CTE levels in humans anesthetized with halothane is reported.


Determination of trimethoprim in biological fluids by high-performance liquid chromatography. A rapid, sensitive, and specific high-performance liquid chromatographic assay was developed for the determination of trimethoprim in blood, plasma, and urine using normalphase (adsorption) chromatography on a microparticulate silica column and UV monitoring at 280 nm. Trimethoprim is selectively extracted from the biological sample matrix at alkaline pH with chloroform, providng nearly quantitative extraction (greater than 95%) and a sensitivity limit of 0.01 to 0.02 microgram/ml blood or plasma, without interference from sulfonamides.


Simple method for the determination of choline and acetylcholine by prolysis gas chromatography. An improved purification procedure is described for the simultaneous assay of endogenous choline and acetylcholine by pyrolysis gas chromatography, particularly for providing a simple and effective method for propionylation of choline in the presence of acetylcholine. The reaction was carried out in acetonitrile solution prepared by dissolving the evaporated residue of the supernatant of brain homogenate. Thus samples for propionylation were prepared without the use of ion-exchange chromatography.


High-performance liquid chromatographic determination of hydrocortisone and methylprednisolone and their hemisuccinate esters in human serum. A high-performance liquid chromatographic method is described for the simultaneous determination of methylprednisolone (MP) and methylprednisolone hemisuccinate (MPHS), or hydrocortisone (HC) and hydrocortisone hemisuccinate (HCHS) in human serum. Reversed-phase liquid chromatography was performed on a microparticulate C18 column (Spherisorb, 5 micron) using a mobile phase of 2% glacial acetic acid, 30--35% acetonitrile, 70--65% water with ultraviolet detection (254 nm). The method uses 17 alpha-hydroxyprogesterone as the internal standard for the determination of methylprednisolone and its hemisuccinate ester, or 11-deoxy-17-hydroxycorticosterone as the internal standard for the determination of hydrocortisone and its hemisuccinate ester. The sensitivity is 0.03 microgram/ml for HC, 0.07 microgram/ml for MP, 0.04 microgram/ml for MPHS, and 0.10 microgram/ml for HCHS, with a detection limit of 0.02 microgram/ml for all four steroids. Calibration curves are linear up to 3 micrograms/ml for MP or MPHS (as equivalent MP) and up to 4 micrograms/ml for HC and 7 micrograms/ml (as equivalent HC) for HCHS. The pooled relative standard deviation for replicate for each steroid is less than 7%. Plasma concentration--time curves are reported for MP and MPHS or HC and HCHS of two human subjects following intramuscular administration of 125 mg of methylprednisolone sodium succinate for injection, U.S.P., or 250 mg of hydrocortisone sodium succinate for injection, U.S.P.


[Identification of 4-O-methyldopamine in rat tissues by reverded-phase liquid chromatography (author's transl)]. 4-O-Methyldopamine was identified and assayed in tissues from L-dopa treated rats by reversed-phase high-performance liquid chromatography. The initial steps in the separation of catecholamines were performed by alumine, a weak cation-exchange resin, and thin-layer chromatographic techniques. After L-[3 H] dopa administration, the radiochromatogram was superimposed on the fluorochromatogram obtained with authentic marker 4-O-methyldopamine. This metabolite was detected in kidney but not in brain. The 4-O-methyldopamine:3-O-methyldopamine ratio was 0.032 in kidney. The influence of various treatments on this ratio was investigated. A 160% increase was found after L-dopa administration. This effect was potentiated by nialamide pretreatment (550% increase).


Determination of the naturally occurring monoacetyl derivatives of di- and polyamines. A method is described for the determination of pmol quantities of monoacetylputrescine, N1-acetylspermidine, N8-acetylspermidine and related compounds. The method is based on the derivation of these compounds with 5-dimethylaminonaphthalene-1-sulphonylchloride, followed by thin-layer chromatographic separation. Cleanup steps allow the application of the method to urine analyses. From the repeated determination of acetylated polyamines in the urine of healthy individuals it can be concluded that these conjugates are the major excretory form of di- and polyamines. The cleanup steps used in this procedure and the method described for the stabilization of 5-dimethylaminonaphthalene-1-sulphonyl derivatives on thin-layer plates are advantageous also for the analyses of total polyamines in urine hydrolysates, and in related applications of the dansylation method.


Simultaneous gas chromatographic determination of lorcainide hydrochloride and three of its principal metabolites in biological samples. A method is described for the determination of the antiarrhythmic drug lorcainide hydrochloride and its three main metabolites in plasma, urine, faeces and tissues from man and animals. The procedure involves the extract of the parent drug, its metabolites and the internal standard from the biological materials at different alkaline pH values, back-extraction into sulphuric acid and re-extraction into the organic phase (heptane--isoamyl alcohol). After silylation of the different phenolic and the N-dealkylated metabolites, analyses were carried out by automated gas--liquid chromatography with electron-capture detection. The method has a sensitivity limit of 5 ng for lorcainide, and 10--20 ng for the various metabolites, per millilitre of plasma. The method was applied to urine, faeces, plasma and tissue samples from man and animals. It was also suitable for automatic sample analysis.


Dynamic cation-exchange systems for the separation of drugs derived from butyrophenone and diphenylpiperidine by high-performance liquid chromatography and applied in the determination of halopemide in plasma. Dynamic (solvent generated) cation-exchange systems for the separation of drugs and main metabolites derived from butyrophenone and diphenylpiperidine (haloperidol, pimozide, halopemide) were investigated. The effect of organic modifier, detergent, counter-ion concentration and of the pH on the retention has been determined. The results show that variation of these parameters permits adjustment of the retention of these drugs over a wide range. The dynamic cation-exchange system developed was applied to the determination of halopemide and its main metabolite in plasma. The precision and detection limit of the method and the extraction efficiency were established. The time course of halopemide and plasma levels of patients chronically receiving halopemide are reported.


Quantitative analysis of sulpiride in body fluids by high-performance liquid chromatography with fluorescence detection. A high-performance liquid chromatographic method for the analysis of sulpiride, N-ethyl-2-(2-methoxy-5-sulphonamido-benzamido-methyl)-pyrrolidine, in body fluids is described. A structurally related compound, N-ethyl-2-(2,4-dimethoxy-benzamido-methyl)-pyrrolidine, was used as internal standard. A fluorescence detector with excitation maximum at 299 nm and emission maximum at 342 nm was used for the quantitation. The detection limit was about 10 ng/ml in serum and cerebrospinal fluid and about 200 ng/ml in urine. The experimental error was 5--10% in the concentration range 25--100 ng/ml. Some preliminary data from a pharmacokinetic study in healthy volunteers are presented. The half-life for sulpiride in serum was about 8 h. Sulpiride was also measured in cerebrospinal fluid from five drug-treated psychotic patients.


[High-pressure liquid chromatographic determination of benfurodil hemisuccinate in blood and urine (author's transl)]. A sensitive and reliable method for quantitative determination of benfurodil hemisuccinate and benfurodil in plasma by high-pressure liquid chromatography on a Zorbax SIL column with a mean particle size of 7 micrometer and UV detection at 254 nm is described. Benfurodil hemisuccinate is stable in plasma but not in aqueous solutions. This is explained by its great fixation to plasma proteins which has been shown by equilibrium dialysis.


Detection of endogenous salsolinol in neonatal rat tissue by a radioenzymatic--thin-layer chromatographic assay. A sensitive radioenzymaticc--thin-layer chromatographic assay for the quantitative analysis of the tetrahydroisoquinoline alkaloid, salsolinol, in plasma and neonatal rat tissue is described. The assay involves the enzymatic O-methylation of salsolinol by catechol-O-methyltransferase in presence of [3H]S-adenosylmethionine, and subsequent separation by thin-layer chromatography of the resultant [3H]O-methyl-salsolinol from the O-methylated derivatives of dopamine, epinephrine and norepinephrine. The method allows the detection of as little as 100 pg salsolinol per g tissue, and the accurate quantitation of as little as 100 pg/ml plasma and 500 pg/g tissue. This assay permitted the detection of trace amounts of endogenous salsolinol in neonatal rat tissue (less than 500 pg/g tissue).


New procedure for isolation of amino acids based on selective hydrolysis of trimethylsilyl derivatives. A rapid procedure for the isolation of amino acids from physiological fluids by class separation suitable for gas chromatographic and gas chromatographic-mass spectrometric analysis is described. A physiological fluid such as plasma is adjusted to pH 2 and extracted with diethyl ether to remove organic acids and neutrals. After precipitation of proteins with trichloroacetic acid, the aqueous plasma is dried and derivatized by trimethylsilylation. Organic compounds like sugars and amino acids are rendered soluble in petroleum ether leaving inorganic salts when the soluble layer is transferred. Separation of sugars from amino acids is achieved by taking advantage of the different rates of aqueous hydrolysis of the trimethylsilyl (TMS) derivatives. Mixing the petroleum ether extract with a small volume of water results in two phases. The petroleum ether layer contains TMS-Sugar constituents of plasma and the aqueous layer contains free amino acids and amines. This procedure was used to isolate L-dopa, 3-O-methyldopa and tyrosine from human plasma in a quantitation assay using 18O-labelled amino acids and gas chromatography-mass spectrometry.


Determination of erythrocyte amino acids by gas chromatography. Erythrocyte amino acid levels were determined, by gas chromatography, in a group of 34 normal human adults. No significant sex or age correlations were noted. A method for the quantitative gas chromatographic analysis of free amino acids in erythrocytes is described. Following hemolysis and deproteinization the amino acids were isolated on a cation-exchange resin. Glutathione was removed from the amino acid mixture by adsorption on an anion-exchange resin. Following conversion to their N-acetyl-n-propyl esters, 19 amino acids were separated and quantitated by gas chromatography on a single column in 18 min. Typical reproducibility data indicate that a coefficient of variation of 2-5% is attainable.


Determination of tryptophan and several of its metabolites in physiological samples by reversed-phase liquid chromatography with electrochemical detection. A new method for the concurrent assay of three tryptophan metabolites at the picomole level is described. The method has been developed for blood, urine, cerebrospinal fluid, and tissue samples such as whole brain, brain parts, and endocrine glands. Tryptophan itself, serotonin, and 5-hydroxyindoleacetic acid are isolated initially on extraction columns, eluted with a suitable solvent, and injected onto a liquid chromatograph with an amperometric detector. This general approach may be applicable to a variety of other tryptophan metabolites and should be useful in both research and clinical investigations.


Determination of therapeutic and toxic concentrations of doxepin and loxapine using gas-liquid chromatography with a nitrogen-sensitive detector, and gas chromatography-mass spectrometry of loxapine. A gas-liquid chromatographic procedure is presented for the determination of therapeutic and toxic serum levels of doxepin and loxapine, using a nitrogen-phosphorus-sensitive detector. Amitriptyline is used as the internal standard. The method is accurate, sensitive and specific with no derivatization required prior to analysis. An advantage of the procedure is the small serum sample size needed for analysis and the selectivity and sensitivity of the detector, with the limit of detection being 3 and 2 microgram/l for doxepin and loxapine, respectively. Nine cases of doxenin and loxapine misuse are presented. Serum doxepin concentrations ranged from 113 to 439 microgram/l, with a loxapine concentration of 192 microgram/l observed in one patient. The presence of the tricyclics was identified and confirmed by gas chromatography-mass spectrometry and the mass spectrum of loxapine is reported.


Measurement of adriamycin (doxorubicin) and its metabolites in human plasma using reversed-phase high-performance liquid chromatography and fluorescence detection. We describe a method for measuring adriamycin and its major metabolite, adriamycinol, in plasma, using reversed-phase high-performance liquid chromatography and fluorescence detection. The lower limit of detection is approximately 1 ng/ml for both compounds; within-day coefficients of variation are 3.6% and 4.4% for adriamycin and adriamycinol, respectively. A slight modification of this procedure also allows measurement of aglycone metabolites.


Reversed-phase liquid chromatographic determination of plasma levels of adriamycin and adriamycinol. A method is given for the determination of adriamycin and its main metabolite, adriamycinol in plasma from cancer patients after administration of adriamycin as the free drug or as a complex with DNA. Adriamycin and adriamycinol are extracted in a column from 1 ml of plasma (pH 8.6) using a mixture of chloroform--1-heptanol (8:2). After re-extraction into phosphate buffer pH 2.2, the separation is performed as reversed-phase liquid chromatography on a LiChrosorb RP-2 (5 micron) column with a mobile phase of acetonitrile-water, acidified with phosphoric acid. The precision by quantitation with photometric detection was better than 5% within the range 50-300 ng/ml. Plasma levels of adriamycin and adriamycinol in a cancer patient are presented in this paper.


Spectrofluorimetric determination of dipyridamole in serum--a comparison of two methods. Two spectrofluorimetric methods for the determination of dipyridamole in plasma are described. The thin-layer chromatographic-fluoridensitometric method utilizes 1 ml of plasma which is extracted at pH 10 with diethyl ether-dichloromethane (80:20). The organic phase is evaporated to dryness, reconstituted in 250 microliter dichloromethane and 5 microliter are spotted on a silica gel 60 plate. The plate is developed in ethyl acetate-methanol-ammonia (85:10:5), dried, dipped in a paraffin wax solution, dried, and scanned using 380 nm as excitation wavelength, a 430 nm cut-off filter, and collecting all emitted light on the photomultiplier. Quantitation was done by the external standard method, peak heights being measured and a calibration graph constructed. For the spectrofluorimetric method 1 ml of plasma is extracted at pH 10 with 8 ml of hexane-isoamyl alcohol (95:5) and the organic phase used directly for the measurement of the fluorescence intensity (excitation 405 nm, emission 495 nm). Quantitation was done by measuring the fluorescence of standards that were treated as above and constructing a calibration graph of concentration versus fluorescence intensity. Concentrations of unknowns were found by interpolation from this graph. The two methods were found to exhibit good correlation but the spectrofluorimetric method proved to be more amenable to the analysis of a large number of samples.


Dual-detector-post-column reactor system for the detection of isoenzymes separated by high-performance liquid chromatography. I. Description and theory. We describe a dual-detector-post-column chromatographic reaction detector system that corrects for substances present in biological samples that interfere with the measurement of isoenzymes separated on a chromatographic column. The response observed at the detector in front of the reaction coil is mathematically dispersed, time transformed and subtracted from the detector behind the coil to produce a blank corrected chromatogram. The same computer program calculates peak areas and other chromatographic parameters such as height equivalent to a theoretical plate and retention time. In addition, we have evaluated the dispersion effects caused by various changes in our experimental system.


Dual-detector-post-column reactor system for the detection of isoenzymes separated by high-performance liquid chromatography. II. Evaluation and application to lactate dehydrogenase isoenzymes. We describe the separation of lactate dehydrogenase isoenzymes by high-performance liquid chromatography-anion-exchange columns and their quantitation by a computer-controlled, dual-detector post-column reaction system. The recoveries from the separation column were ca. 90%. The dynamic range of the system was linear over about three orders of magnitude from 3 to 1500 U/l. The coefficient of variation for isoenzyme peak areas was ca. 2%. The method is compared to the classical electrophoresis measurement and shows increased speed, resolution, precision and accuracy.


Determination of bromhexine as its trifluoroacetyl derivative by gas-liquid chromatography with electron-capture detection. A specific gas-liquid chromatographic method with electron-capture detection is described for the determination of bromhexine at the nanogram level. Structurally analogous internal standards were synthesized and their suitability was investigated, based on their chromatographic properties on different stationary liquid phases. The electron-capture activity of the compounds is increased by trifluoroacetylation. Reaction conditions for this derivatization were studied. Calibration graphs in the range 0-57 ng (in a total of 100 microliters of reaction mixture) showed good linearity.


Screening methods for the detection of thirteen common mycotoxins. A study of screening methods for thirteen mycotoxins showed that they can be separated as neutral and acidic metabolites. RF values were determined in several solvent systems. The reactions of the mycotoxins with well known spray reagents were investigated, and their detection limits were established. A general procedure for the extraction of mycotoxins from contaminated samples is described.


Separation of tryptic peptides of normal and abnormal alpha, beta, gamma, and delta hemoglobin chains by high-performance liquid chromatography. High-performance liquid chromatography (HPLC) was used to separate tryptic peptides of the normal alpha, beta, gamma, and delta chains of human hemoglobins A, F, and A2 and of the abnormal chains of 25 hemoglobin variants. In addition, the separation of chymotryptic peptides of the oxidized core of the normal alpha chain by HPLC was evaluated. HPLC has several advantages over conventional methods used for the separation of proteolytic fragments of hemoglobin chains. The method is fast, and reproducible, and requires only small quantities of material. Several peptides are eluted as single zones, thus eliminating the need of rechromatography for further purification. Characteristic changes in the elution pattern of the peptides often indicate specific modifications.


High-performance liquid chromatographic determination of the nitroimidazole azanidazole in human plasma and urine. Azanidazole can be measured in plasma and urine by reversed-phase high-performance liquid chromatography employing UV detection. Peak mean plasma concentrations of azanidazole, of 267 ng/ml, occurred at 1.5 h after single oral doses to human subjects, and declined with a half-life of 0.8 h. Less than 0.5% of the dose was excreted in the urine as unchanged drug. Metabolites of azanidazole were also detected by the procedures used.


Improved thin-layer chromatographic method for sugar separations. A method is described for the utilization of pre-coated, non-impregnated silical gel thin layers in one- and two-dimensional separations of carbodhyrates and related compounds. Boric and phenylboronic acids were added to the organic elution systems in different concentrations and their interactions with the sugar molecules during the chromatographic process were studied. A comparison was made between solvent systems containing boric or phenylboronic acid and systems devoid of both acids as eluents. With boric acid-containing solvents the migration of some sugars was considerably inhibited, whereas phenylboronic acid produced an increase in the RF values of certain sugars. The combination of these two types of solvent in two dimensional development resulted in the clear separation of a group of mono- and disaccharides of bioclinical interest.


Equilibrium saturation chromatographic method for studying the binding of ligands to human serum albumin by high-performance liquid chromatography. Influence of fatty acids and sodium dodecyl sulphate on warfarin-human serum albumin binding. A size exclusion chromatographic method for studying ligand-macromolecule binding parameters is described. This equilibrium saturation method allows the determination of the concentrations of constituents in equilibrium and is specially useful for characterizing ligand--protein binding under conditions that can be compared with physiological conditions. The method has been used for measuring warfarin--human serum albumin (HSA) binding and for studying the influence of free fatty acids (FFA) and sodium dodecyl sulphate on warfarin--HSA binding. Some comparisons with the Hummel and Dreyer method are given. The influence of the FFA is strongly dependent on their chain length, with an inversion of the effect for a 10-carbon chain.


Methylthiohydantoin amino acids: chromatographic separation and comparison to phenylthiohydantoin amino acids. Most phenylthiohydantoin (PTH) amino acids and most methylthiohydantoin (MTH) amino acids may be separated from one another by thin-layer chromatography (TLC) using the same sequential development technique with the same two solvents. Similarly, a single solvent system may be used in high-performance liquid chromatography (HPLC) to separate most PTH-amino acids and most MTH-amino acids. When both TLC and HPLC separations are performed on a sample, all MTH-and PTH-amino acids can be uniquely identified. Since many solid-phase protein sequencing techniques generate both MTH-and PTH-amino acids, these analytical systems simplify identification of the amino acid derivatives. Although the chromatographic properties of MTH-and PTH-amino acids are similar, they are not identical (contrary to a previous report).


Simple method for the simultaneous determination of noradrenaline, dopamine and serotonin by stepwise elution from a short column of weak cation-exchange resin. A simple method for the simultaneous determination of noradrenaline, dopamine and serotonin using a short column of Amberlite CG-50 is described. Noradrenaline and dopamine were eluted from the column with phosphate buffers containing 1.5 and 4.0% boric acid, respectively, and then serotonin was eluted with 1.0 N HCl. Catecholamines were determined by a modification of the ethylenediamine condensation method using potassium ferricyanide as oxidant and isobutanol for extraction of the fluorophores. Serotonin was measured by the acidic o-phthalaldehyde method. The method was applied to the simultaneous determination of noradrenaline, dopamine and serotonin in discrete regions of rat brain.


Gas chromatographic-mass spectrometric analysis of formaldehyde in ambient air using a sampling tube. A gas chromatographic-mass spectrometric technique for the analysis of trace concentrations of formaldehyde in air is described. Molecular Sieve 13X was found to be an excellent adsorbent. The collected samples were thermally desorbed onto the analytical column (Porapak T) for separation, and quantified by mass fragmentography (m/e 29 and 30). Advantages of the technique include ppb sensitivity, selectivity and quantitative recovery. Experimental results are given for air samples in a rural area.


Methylation analysis in glycoprotein chemistry. General procedure for quantification of the products of solvolysis of permethylated glycopeptides and glycoproteins. A technique for the gas chromatographic analysis of the products of solvolysis of permethylated glycopeptides and glycoproteins has been developed. It involves methanolysis of a permethylated compound, quantitative transformation of methyl ethers of methyl glycosides into the corresponding O-trimethylsilyl(TMS)-O-methylalditols [2-deoxy-2-(N-methyl)acetamido-O-TMS-O-methylalditols in the case of hexosamine derivatives] and gas chromatographic quantification using a single column packed with 0.4% OV-225 on surface-modified Chromosorb.


Season and treatment effects on serum prolactin and milk yield during induced lactation. Nineteen nonpregnant, nonlactating dairy cows were allotted to three treatments to induce lactation during winter, 1976, or spring, 1977. All groups received 17 beta-estradiol (.1 mg/kg) days 1 to 7. Groups 2 and 3 also received progesterone (.25 mg/kg) days 1 to 7. Groups 1 and 2 were given reserpine (5 mg intramuscular) on days 8, 10, 12, and 14. Group 3 received reserpine (5 mg intramuscular) on days 2, 5, 8, 11, and 14. Blood samples were collected for prolactin analysis just prior to and 3 h after reserpine injection. Mean daily temperatures were 11.9 C for spring group and -6.5 C for winter group. Comparisons of spring with winter for basal prolactin concentrations, reserpine-stimulated prolactin concentrations, and 100-day milk yields were 44 with 10 ng/ml, 482 with 199 ng/ml, and 1991 with 862 kg. Differences in prolactin concentrations and milk yields among hormone and reserpine treatments could not be detected, but cows on treatment 3 in the spring gave the largest yield of milk. Prolactin concentrations were correlated with milk yields among cows and among cows within seasons. Seasonal differences demonstrate the critical role of prolactin in the treatment to induce lactation.


Effects of low chloride intake on performance, clinical characteristics, and chloride, sodium, potassium, and nitrogen metabolism in dairy calves. Young male Holstein calves were fed either a control (.5% chloride) or a low-chloride (.038% chloride) practical diet for 7 wk. Both groups received low-chloride (.00038% chloride) well water. Feeding the low-chloride diet did not produce definite clinical symptoms of chloride deficiency. Neither body weight gains, feed intake, feed digestibility, nor body retention of chloride, sodium, potassium, or nitrogen were effected adversely. Although the chloride intake of the low-chloride calves was only one-sixteenth that of controls, body chloride retention was similar for the two groups. The similar retention of body chloride was due to effective homeostatic mechanisms in which urinary chloride excretion was reduced by 95% in the low-chloride calves. Low-chloride calves consumed more water and excreted more urine than control calves. Although the exact minimum chloride requirement for growth in calves was not established, .038% chloride was adequate for normal growth for the 7 wk.


Colostral immunoglobulin transfer in calves. III. Amount of absorption. Amount of colostrum fed and age at first feeding are the two major factors in determining maximum immunoglobulin concentration in serum for each immunoglobulin class in postcolostral calves. Both factors show linear response: increasing age, decreasing concentration; increasing amount fed up to 2 liters, increasing concentration. The two factors interact in a curvilinear response. Calves with initial feeding at progressively older ages need less colostrum to reach maximum absorption. Maximum absorption is represented by a progressively lower immunoglobulin concentration in serum with increasing age. Response surface contours for the maximum immunoglobulins IgG, IgM, and IgA, based on experimental data, have been included. Neither body weights of the experimental calves nor pooled colostral concentrations of immunoglobulin from pooled colostrums fed to the calves influenced maximum concentrations of immunoglobulin in serum.


Health care requirements of dairy cattle. I. Response to milk yield selection. To measure direct response to single trait selection for milk yield and correlated response in health problems, two homologous base populations were formed by pairing 66 Holstein females by sire. Base populations and descendants were managed identically except for selection by milk yield. One base and lineage were mated with sires with highest predicted difference milk (selection group); other base and lineage were mated with average sires in 1964 (control group). Milk yield, supplemental labor, and veterinary and semen expense were recorded specific to each cow. Over 9 yr, 130 selection and 163 control cows were observed. Selection cows yielded more milk but with increase in labor and expense for health care. Estimates of labor for the selection group were greater for mammary, locomotion, and digestion categories and for expense in mammary, respiration, first insemination, and later insemination categories. Labor and expense for reproduction did not differ for genetic groups. Differences between groups in annual estimated labor and expense totaled $27.00. Extra income over feed cost more than compensated for greater health care and semen cost; however, the magnitude of the latter indicates a need for them to be considered when economically evaluating breeding programs where major emphasis is on milk yield.


Three times a day milking during the first half of lactation. Three milking frequency treatments were compared: twice daily milking; thrice daily milking until milk dropped below 24 kg; thrice daily milking until milk dropped below 31 kg. Three time milking was at least 45 days but no more than 150 days. Cows (12 to 14 per group) were managed alike except for milking frequency. In early lactation, increased milking had little effect. With time the superiority in yields increased such that cows on three times for 150 days were outproducing two time cows by 20%. Cumulative milk yields were greater for the thrice groups than for the twice group by 5% at 56 days, 11 and 8% at 154 days, 11 and 9% at 182 days, and 10% at 280 days. Fat percentage, adjusted for previous lactation fat percentage, averaged .2 to .3% lower for the cows milked three times until 24 kig but only .1% lower for cows milked thrice until 31 kg. Increased yield was primarily from prolonged peak yield and less subsequent decline. Switching from three to two milkings decreased yield 6 to 8% in the 1st wk. However, three time milking had a positive carryover, apparently due to higher starting yield at the point at which they were switched.


[Fossae, recesses and culs-de-sac in the pelvic peritoneum of women (author's transl)]. The pelvic peritoneum in women is peculiarly irregular because of the numerous folds, fossae, recesses and culs-de-sac that exist in it. Blood and purulent serous fluids occur in the abdomino-pelvic cavity and stagnate in these sites. Since they are particularly to be found around the adnexae they can give rise to secondary infection occurring as a result of primary adnexal infection. Furthermore, when serous fluid is regurgitated into the tubo-ovarian hollow through the abdominal os of the tube and into the pertoneum around the ovary, adhesions and endometrial deposits occurring in this area can be explained. Each one of these structures should be systematically explored, particularly when hystero-salpingographies and laparoscopies are being carried out. Furthermore, they should be cleaned out very meticulously after all conservative surgery to prevent the formation of adhesions.


[Maternal mortality in the gynaecological and obstetrical clinic of the University Hospital in Rennes. A retrospective study over 10 years, from 1969 to 1979 (author's transl)]. Maternal mortality in the C.H.U. of Rennes from the 1st January 1969 to the 1st January 1979 consisted of 8 deaths in 24.911 deliveries and 1.381 Caesarean sections, which gives a mortality of 32 per 100.000 deliveries. There was 1 death due to hyperemesis gravidarum, 1 due to toxaemia of pregnancy, 3 due to haemorrhage, 1 due to puerperal sepsis, 1 due to cerebral haemorrhage and lastly 1 due to cancer of the pancreas. 2 women died during Caesarean section, which works out at 1 death per 690 Caesareans. Of these 8 deaths, 4 were possibly avoidable, and 3 of their children survived. In spite of continual progress since the end of the second world war, maternal mortality should decrease still further and tend down to the figures for Scandinavia or England (10 deaths for 100.000 deliveries).


[Arterio-venous fistulae of interest in the uterine vessels (author's transl)]. The authors report an interesting case of arterio-venous fistula in the uterine vessels in a young two-para of 23 years of age. A pulsatile and throbbing mass was found in the right parametrium at her post-natal visit after the second delivery, although no abnormal sign had been noted during the pregnancy. The diagnosis was confirmed by arteriography. Two surgical procedures have not been able to cure this patient entirely of these multiple fistulae which were congenital in origin. The production of an arterial embolus finally got over the difficulty of removing the residual troubles at the same time allowing the patient to keep her uterus. Following this case, the authors discuss the aetiology of acquired or congenital fistulae and the means by which this rare vascular pathological condition can be diagnosed and treated in the pelvis.


[Vulvar granuloma due to Enterobius vermicularis (author's transl)]. The authors report a case of casual discovery of a enterobiasic vulvar granuloma arising on the left labia. This woman, living in Lille (France), 70 years had been treated by surgery and roentgentherapy for a well differentiated squamous carcinoma of the vulva. They consider the question of the genital localisation of Enterobius vermicularis in women and discuss the pathogenic hypothesis.


[A computerised study of 124 cases of varicocoele which were operated on for diminished fertility. General features (author's transl)]. A computerised study has been carried out on the general features of a clinical, biological and histological nature occurring in 124 varicocoele cases that were operated. These features were furthermore studied in correlation with the degree of varicocoele in existence and whether characteristics were found in isolation or in association with one another. The authors conclude that there were significant correlations between the degree of varicocoele present and the hormonal and histological findings. They advance a theory about the transport of testosterone in the spermatic cord which can explain the low level of testosterone found in the blood in these varicocoeles.


[Neonatal screening for congenital hypothyroidism by measuring TSH in dried blood samples. Two years experience in the method (author's transl)]. Systematic screening for congenital hypothyroidism was started in Lyon in September 1976. This screening was coupled with PKU screening, using the same dried blood samples in filter paper obtained on the 5 th day of life. TSH levels were determined by radioimmunoassay adapted for dried blood samples (Kit Abbott). In 24 months, 56 176 samples were analyzed. The critical level calling for control was successively raised from 20 to 30 now 40 muUl/ml of serum. A high level of TSH was found in 307 children (0.55 p. 100). Pathological deliveries were found in most of these infants (neonatal injury, cesarean section, forceps or ocytocic perfusion, neonatal icterus) and a second or a third measurement showed normal TSH level. Congenital hypothyroidism, was detected in 18 infants: 12 ectopic gland, 5 athyreosis and 1 dyshormonogenesis. Treatment was begun at a mean age of 38 days (29 to 50 days). Despite a short follow-up the psychomotor development of the infants seems to be normal in all cases but one (one athyreosis with a neonatal injury and a malformative syndrome).


[Clindamycin levels in the cervical mucus after intramuscular administration of the drug (author's transl)]. Female genital tract is a major area of anaerobic infections. Clindamycin has been used as a highly active antibiotic against both gram positive and negative anaerobic pathogens including bacteroides fragilis. Concentration of clindamycin in cervical mucus has been tested in 30 women after administration of one single dose (600 mg) of the antibiotic intramuscularly. Antibiotic levels were tested by agar diffusion method employing a strain of alpha-hemolytic streptococcus. Levels of clindamycin in cervical mucus were found to be sufficient to inhibit "in vitro" a significant proportion of anaerobic bacteria present in cervical mucus.


[Severe epistaxis during pregnancy. A case history (author's transl)]. We report on a case of severe epistaxis in pregnancy with maternal collapse and fetal death. A few similar cases have been published in the literature. These epistaxis happen towards the end of the pregnancy and occur with hyperaemia of the nasal mucosa, which is very probably hormone dependent. Delivery or death of the fetus brings about immediate cessation in the bleeding. The mother's life and that of the fetus can be jeopardised because of the severity of the bleeding or its recurrence, which makes it a very severe condition. When local treatment is useless in such a case two radical treatment have to be considered: the one is surgical, and is ligature of the sphenopalatine artery, and the other is obstetrical, and is termination of the pregnancy (which brings to an end the aetiological factor).


A comparison of the characteristics of two strains of Schistosoma intercalatum Fisher, 1934 in mice. Schistosoma intercalatum from Cameroon and Zaire each have their specific snail intermediate host. Hybridization experiments between S. intercalatum from Cameroon and Zaire have resulted in viable miracidia for only two generations. If the two strains are of the same species, cross breeding should be possible for several generations. This paper presents a comparison of the characteristics of the two strains of S. intercalatum in experimental infections in mice. Worm return, male-female worm ratio, growth rate, number of testes in male adult worms are compared. The quantitative distribution of eggs in the tissues of the mice is recorded. A comparison of the size and shape of the parasite eggs is described as are the changes in the weight of the liver and spleen of the infected mice. It is considered that the differences observed between the two strains of S. intercalatum do not necessitate a division of the two strains into two separate species.


[Prostaglandin F-2-alpha gel for induction of midtrimester abortion (author's transl)]. Prostaglandin F2-alpha was administered extra-amniotic as a gel through the cervix uteri to 100 patients in the 2nd trimester of pregnancy: 82 for termination of pregnancy, 17 for intrauterine death, and 1 for a hydatiform mole. The success rate was 96 p. 100. Mean duration of induction was 17 hrs 06 min., cervical priming lowered this duration to 11 hrs 40 min. (p less than 0.01). Extradural analgesia provided satisfactory pain relief and allowed a dilatation and curettage to be performed without recoure to a general anaesthetic. The technique described showed a lower incidence of side-effects than other modes of administration of PG F2-alpha, and proved to be effective, reliable, and acceptable to patients.


Studies of the relationships between Schistosoma and their intermediate hosts. II. The genus Bulinus and Schistosoma haematobium from Sudan, Zaire and Zambia. The relationship between Schistosoma haematobium from Sudan, Zaire and Zambia and various species and strains of Bulinus, has been investigated. The main emphasis was placed on evaluating the total cercarial production per 100 exposed snails as an index of the compatibility between snail and schistosome. The observations confirmed the results of previous compatibility studies which showed that there were two groups of S. haematobium, truncatus-borne from Sudan and Zaire, and africanus-borne, from Zambia. They also revealed that this division was not absolute, as some overlapping occurred. However, if the cercarial production was taken as an indication of compatibility, instead of the infection rate, there was a more definite division between the two groups. The observation that there is a truncatus-borne strain of S. haematobium present in Zaire extends the area where there is a mixture of truncatus- and africanus-borne strains farther southwards.


Vitamin absorption in the in vivo intestine of normal and infected (Hymenolepis diminuta: Cestoda) rats. Uptake and serosal transfer of the vitamins thiamine, riboflavin and folic acid have been studied in vivo in normal and parasitized rats infected with Hymenolepis diminuta (Cestoda). Regional differences in intestinal uptake of all three vitamins in both uninfected and parasitized animals were not satistically significant. In the parasitized intestine mucosal uptake and serosal transfer of thiamine were significantly inhibited, with increased mucosal accumulation of the vitamin as luminal thiamine concentration increased. Apparent increased riboflavin mucosal uptake in parasitized animals, was not matched by the reduced serosal transfer, suggesting adsorption of the vitamin in the unstirred aqueous layers. Mucosal uptake of folic acid increased in the parasitized gut; serosal transfer and mucosal accumulation were not affected. These results, indicating vitamin malabsorption associated with infection by H. diminuta, are consistent with the parasite inhibiting mucosal passive transport mechanisms. This conclusion is supported by the changes in net water fluxes associated with vitamin uptake in the parasitized intestine.


The serodiagnosis of human hydatid disease: 1. The routine use of latex-agglutination and complement-fixation in diagnosis. The combined use of complement fixation (CF) and latex agglutination (LA) tests is reported on sera from 6328 patients with suspected hydatid disease; 191 were confirmed positive at operation ('known positives'). Results by LA are related to CF titres. Both tests were negative in 90% of specimens. Nine patients were subsequently found infected of whom 3 became positive in tests after operation. Of sera positive in both tests, 75% were from 'known positives'. The remainder were almost certainly from infected patients. Half the patients whose sera were LA positive/CF less than or equal to 1/4 were follow-up 'known positives' in whom CF titres had waned; 2 were early infections. Only 3% of the cases with an LA negative/CF titre of greater than or equal to 1/16 were 'known positives' and 6% where the CF titre was 1/8. The remaining CF results in the group were false positives and accounted for 1.2% of all sera tested. Findings show that a CF titre greater than or equal to 1/8 with positive LA indicates past or present infection; a negative CF test with positive LA usually indicates past infection; rarely, infection is present when a CF titre is greater than or equal to 1/8 and LA is negative. A rising CF titre and positive LA indicates current infection; reliable prognosis following treatment is given by CF.


Studies of the relationship between Schistosoma and their intermediate hosts. III. The genus Biomphalaria and Schistosoma mansoni from Egypt, Kenya, Sudan, Uganda, West Indies (St. Lucia) and Zaire (two different strains: Katanga and Kinshasa). The compatibility between strains of Schistosoma mansoni from Egypt, Kenya, Sudan, Uganda, the West Indies, and Zaire (two strains which came from Katanga and from Kinshasa), and various species and strains of Biomphalaria, i.e. Biomphalaria pfeifferi, B. alexandrina, B. glabrata and B. camerunensis was investigated. Data as mortality, rate of infection of the surviving snails, duration of infection, cercarial production per day per positive snail, etc., were observed. The main emphasis was placed on determining the total cercarial production per 100 exposed snails for each snail population. It was possible to infect all the tested populations of B pfeifferi with the various strains of S. mansoni, but the observation as e.g. TCP/100 exposed snails varied greatly according to the population of snail and the strain of S. mansoni. The results for the remaining species of Biomphalaria varied greatly, depending on the combination, e.g. B. alexandrina was only susceptible to the local S. mansoni from Egypt. The highest TCP/100 exposed snails was more than 1 million for the strains of S. mansoni from Egypt, Kenya and the West Indies in B. alexandrina, B. pfeifferi and B. glabrata, respectively. The next group, with a TCP/100 exposed snails on 7--800 000 consists of S. mansoni from Sudan, Uganda and Zaire (Katanga) all in B. pfeifferi. The last tested strain of S. mansoni, Zaire (Kinshasa) yielded a cercarial production on 500 000 per 100 exposed snails in B. pfeifferi and B. camerunensis. The shortest prepatent period, 19 days, was observed for S. mansoni from Kinshasa, Zaire, in B. camerunensis, and the longest prepatent period, 25 days, was found for strains from Egypt and from the West Indies in B. alexandrina and B. glabrata, respectively. In general, a very long duration of infection, lasting up to 200 days, was observed.


Studies of the relationships between Schistosoma and their intermediate hosts. IV. The genus Bulinus and Schistosoma bovis from Morocco. Schistosoma bovis from Morocco was used in infection experiments with several populations of Bulinus truncatus. The snails from Libya, Malawi, Morocco and Senegal were very compatible with the schistosome since the infection rates were approximately 90%, the mortality was low, and a very high production of cercariae, approximately 1.2 million per 100 exposed snails, was observed. Only a very few B. truncatus (2n = 36) from Rhodesia became infected. B. permembranaceus and B. forskalii were refractory.


Species of Cloacina Linstow, 1898 (Nematoda: Strongyloidea) from the black-tailed wallaby, Wallabia bicolor (Desmarest, 1804) from eastern Australia. Species of Cloacina Linstow, 1898 (Nematoda: Strongyloidea) are reported and/or described from the stomach of the macropodid marsupial Wallabia bicolor (Desmarest, 1804) collected in eastern Australia. Nematodes were examined from 26 hosts collected in 3 states and the number of species of Cloacina in any one host was 2--7. The following species were encountered: C. wallabiae Johnston and Mawson, 1939, C. gallardi Johnston and Mawson, 1940, C. edwardsi Mawson, 1972 and C. cornuta (Davey and Wood, 1938) the last named constituting a new host record. The following new species are described: C. mawsonae sp. nov., C. papillata sp. nov., C. annulata sp. nov., C. castor sp. nov. and C. pollux sp. nov.


Reinfection with influenza A (H3N2) virus in young children and their families. The frequency and consequences of reinfection with influenza A virus were studied by longitudinal observation of families for a three-year period in which two epidemics of influenza A (H3N2) occurred. Seven children followed from birth were reinfected 10-25 months after their first infection. Two children were reinfected by the same H3N2 virus while the others were reinfected with a closely related variant. At least five of these reinfections were accompanied by respiratory illness. Reinfection illness was similar to that accompanying primary infection. For children in the second and third year of life during the 1978 epidemic, the rate of infection was the same for those who had been previously infected (seven of 12) as for those who had not been previously infected (22 of 40). Reinfection was detected in 26% of older siblings and 6% of parents. The occurrence of reinfection may have important implications for elucidation of the protective immune response and for development of prophylaxis for influenzal infections.


Secretory immunity in influenza.

Secretory immunity in influenza. The dynamics of secretory antibody formation, the duration of secretory antibody preservation, and changes in the concentration of secretory antibodies to antigens other than influenza virus were studied in 64 patients with influenza A, 105 patients with influenza B, and 23 persons who had had influenza A. Severe forms of influenza A were accompanied by antibody accumulation in sera and nasal secretions; in some cases of mild forms of this infection, this process was limited by the humoral immunity system. In the first days of severe forms, transudation of antibodies from sera to nasal secretions was noted. Secretory antibodies to influenza A virus were preserved at titers of greater than or equal to 1:4 for four to eight months in persons with mild forms of the disease and for more than eight months in those with severe influenza A complicated with pneumonia. Decreases in the titer of antibodies to agents other than influenza A virus, including influenza B virus, respiratory syncytial virus, adenovirus, and staphylococcus toxin, were demonstrated in association with rises in titers of antibody to influenza A virus. Among patients with influenza B, who were infected with a new influenza virus variant, the formation of circulating antibodies was more intensely stimulated than was the formation of secretory antibodies. No correlation between the level of IgA and the antibody titer in nasal secretions was found.


Activity of human blood leukocytes against Toxoplasma gondii. Previous reports on the ability of Toxoplasma gondii to multiply within human mononuclear phagocytes have supported the concept that the phagocytes may protect Toxoplasma from antibody and act as a transport mechanism for dissemination of the organism throughout the body. However, after phagocytosis of viable Toxoplasma, greater than 80% of freshly isolated human peripheral blood monocytes and 50% of polymorphonuclear leukocytes rapidly destroyed these intracellular organisms. Virtually no replication of Toxoplasma occurred within these cells, as determined by microscopic examination and by uptake of [3H]uracil. These results suggested that circulating phagocytes from humans restrict rather than facilitate the initial dissemination of Toxoplasma.


Infection of untreated primary human amnion monolayers with Chlamydia trachomatis. For development of a laboratory model for chlamydial infection of the human genital tract, untreated primary human epithelial cell monolayers, prepared from each of four placentas, were infected with a stock strain of Chlamydia trachomatis isolated from the nasopharynx of an infant with pneumonia. Infectivity titrations were performed with use of standard procedures of chlamydial inoculation, and inclusions grown in amnion cell monolayers and in McCoy cells were measured versus infectious units inoculated. By linear regression analysis, untreated human amnion cells (y = 0.948x + 4.63, r = 0.9255) were as susceptible as McCoy cells treated with diethylaminoethyl dextran and 5-iodo-2-deoxyuridine (y = 1.104x + 5.35, r = 0.9804). Human amnion may prove a valuable model for study of human chlamydial infection and may have important implications for investigation of clinical disease.


Hepatitis B virus infections among Danish surgeons. Acute viral hepatitis B is considered to be an occupational risk to health-care workers worldwide. At the 1977 annual meeting of the Danish Society of Surgeons, 224 participants were examined for the presence of hepatitis B surface antigen (HGsAg) and antibody to HBsAg (anti-HBs) with use of a radioimmunoassay. None of the surgeons was HBsAg-positive, but 23% had anti-HBs. The prevalence of anti-HBs was five times higher among these surgeons than that in an age-matched control population. Hepatitis acquired during occupation as a surgeon was predominantly of type B, in contrast to hepatitis acquired before entering the surgical profession. Danish surgeons must be regarded as a group at high risk of hepatitis B infection.


Single-dose metronidazole and tinidazole as therapy for giardiasis: success rates, side effects, and drug absorption and elimination. Eighty-five symptomatic patients with parasitologically confirmed, recently acquired giardiasis were treated in a comparative trial of 2.4 g of metronidazole either once or on two successive days or 2.0 g of tinidazole once. The follow-up period was eight weeks; the parasitological follow-up consisted of 871 stool and 30 duodenal specimens. Reinfections were unlikely. The rates of success were: metronidazole, single dose, 13 of 26; metronidazole, two doses, 24 of 31; and tinidazole, single dose, 26 of 28. Clinical and parasitological effects were parallel in nearly all cases. Tinidazole was more effective, produced fewer side effects, and was recommendable as the drug of choice in single-dose therapy. Pharmacokinetic explanations for therapeutic failure was sought with use of a bioassay of drug concentrations in serum. The outcome of therapy was not related to serum levels at 1hr or 24 hr, or to the rate of elimination. The mean serum half-lives of active metronidazole and tinidazole were 9.5 and 13.0 hr, respectively.


A model for population regulation with density- and frequency-dependent selection. The life-cycle of a species with separate generations is divided into a 'reproduction phase' and a 'growing-up phase'. In the reproduction phase we assume random mating and selection due to genotype differences in fecundity of the parents and viability of the offspring. During the growing-up phase we assume a (deterministic) death process in continuous time with death rates for the genotypes which increase linearly with the genotype population sizes. In the absence of genotype differences the model gives logistic population regulation. With genotype differences the model generalizes the usual separate generations selection patterns. In addition to these we exhibit cases with three polymorphic equilibria or with a stable cycle.


Delays in physiological systems.

Delays in physiological systems. In comparison to most physical or chemical systems, biological systems are of extreme complexity. In addition the time needed for transport or processing of chemical components or signals may be of considerable length. Thus temporal delays have to be incorporated into models leading to differential-difference and functional differential equations rather than ordinary differential equations. A number of examples, on different levels of biological organization, demonstrate that delays can have an influence on the qualitative behavior of biological systems: The existence or non-existence of instabilities and periodic or even chaotic oscillations can entirely depend on the presence of absence of delays with appropriate duration.


Deterministic single-locus density-dependent selection. Density-regulated selection is considered for a single, multiallele gene locus and separated generations. Characteristics resulting from the basic assumption that the average population fitness decreases with increasing density are derived. Under this assumption, it proves to be necessary to distinguish between regions of allelic frequencies which imply limited population growth, unlimited growth, or ultimate extinction when the population stays in the respective region. Particular attention is given to the investigation of the region of limited growth and the 'carrying capacity function' theta defined on it. Relationships between theta and the average fitness (adaptive surface) in the non-density dependent model are explained. Besides stability properties of equilibrium points, more general characteristics concerning the asymptotic behavior of population trajectories are treated. In this context, the problems of sudden loss of alleles and of population extinction as a result of large fluctuations in density are discussed.


Unlinked strands as a topological constraint on chromosomal DNA, plasmid integration, and DNA repair. It is proposed that circular chromosomal DNA must be constrained such that the two strands are topologically unlinked. This structure can be replicated without strand breakage (nicking) by locally acting enzymes. The recently discovered form V DNA satisfies this topological constraint. The structure is likely to consist of left-handed and right-handed segments of double helix. The constraint of zero linkage has to be preserved by DNA repair, plasmid insertion and by crossing over. The argument presented in this paper is a topological one, following W.F. Pohl, that linkage is a global property that cannot be measured by locally acting enzymes. In contrast to Pohl no argument in favor of a side-by-side structure is presented. A zero linkage constraint would be hereditary and compatible with a multitude of local structures.


Input-output relationship of the Leaky-integrator neuron model. This paper presents a method of calculating the spike sequence at the output of the Leaky-Integrator Neuron Model (LIM) in response to an arbitrary input stimulus. The calculations have revealed new properties of the initial transient behavior of the LIM, as well as new constraints upon necessary and sufficient conditions for the appearance of spikes with a fixed phase relation to a periodic input. It is also possible to infer what knowledge about the input stimulus can be obtained from a temporal sequence of spikes at the output of the LIM. In the Discussion, neuronal examples are considered which do not encode the transmitted information as a spike rate, but rather monitor the time of occurence of individual spikes by comparison with a reference signal. This relatively common case is not adequately treated by previous descriptions based on system theory; ways are suggested by which the formalism developed here can be used to describe completely, and understand more fully, the performance of such systems.


Influence of dietary protein restriction on ovulation, fertilization rates and pre-implantation embryonic development in mice. Eggs were recovered from 153 adult female mice fed either a normal (27% protein) or a protein-deficient (8% protein) diet from day 15 (Group I) or 30 (Group II) before mating until sacrifice. The females were mated with fertile males and sacrificed 48, 72, 96 or 120 hours after observation of vaginal plugs. The study was performed with females that ovulated spontaneously and with females induced to ovulate by injection of pregnant mare's serum gonadotrophin and human chorionic gonadotrophin. Without gonadotrophin treatment, significantly fewer eggs were recovered from malnourished than from normal females. However, after hormonal treatment, the number of eggs ovulated by malnourished and normal females increased similarly. About 50% of the eggs recovered from malnourished females in both groups I and II were cleaved whereas 90% of the eggs recovered from well-nourished females were cleaved. The development of the cleaved eggs from protein deficient females was delayed and asynchronous in comparison with that of eggs from normal females. Embryos recovered from malnourished females also were retarded in differentiation of morulae to blastocysts. It is concluded that a protein-restricted diet adversely affects the fertilization of eggs as well as their early development and differentiation.


Fertilization of hamster eggs in vitro at sperm:egg ratios close to unity. Hamster epididymal spermatozoa were washed and preincubated at extremely low sperm concentrations (100/ml or less) in a culture medium containing naturally-occurring sperm motility-stimulating substances. These substances were partially purified "sperm motility factor" (SMF) derived from hamster adrenal glands and catecholamines (epinephrine or isoproterenol). After preincubation for three hours, a small number (5 or less) of washed, cumulus-free hamster eggs was added to each sperm suspension. Many of these eggs were undergoing fertilization when examined two to three hours later. Fertilization was accomplished in vitro at sperm:egg ratios approaching 1:1, a situation comparable to that believed to exist in vivo. It appears that this demonstration will considerably enhance the potential of in vitro fertilization studies for providing useful information on mammalian gamete interactions.


Polyspermy-preventing mechanisms in mouse eggs fertilized in vitro. The time sequence of the mechanisms that prevent polyspermy in cumulus-free mouse eggs was determined by continuous micropscopic observations during in vitro fertilization. The zona reaction had begun, and the vitelline surface block to polyspermy was established, in less than one minute following sperm fusion with the vitellus. In less than five minutes, the zona reaction had advanced to the stage that no more spermatozoa could become firmly attached to the surface of the zona pellucida.


Failure of human spermatozoa to penetrate zona free mouse and rat ova in vitro. When incubated for 8 to 26 hours with zona-free mouse or rat ova, human spermatozoa failed to attach to or penetrate any of the ova. The ova were capable of being fertilized since both intra- and inter-species penetration of spermatozoa and formation of pronuclei occurred between rat and mouse gametes. When mouse spermatozoa were incubated for three to eight hours with rat ova, a high proportion of the ova were penetrated, formation of pronuclei occurred and in 9 out of 36 ova incubated for 40 hours after insemination, regular cleavage and formation of morphologically normal 2-cell embryos occurred. Human spermatozoa retained their morphological integrity and motility only when the culture medium contained purified bovine serum albumin (3 mg/ml) or human serum (5% v/v) and not when unpurified BSA from several different commercial sources was used as a protein source. In this latter medium, the ova of both rats and mice degenerated after 8-hour incubation in the presence of human spermatozoa but not when human spermatozoa were absent or in the presence of either rat or mouse spermatozoa. Electron microscopy indicated that the human spermatozoa incubated for eight hours in medium containing purified BSA had undergone an acrosome reaction. These spermatozoa also attached to and penetrated human oocytes which had been matured in vitro.


Beat frequency is bimodally distributed in spermatozoa from T/t12 mice. Flagellar beat frequencies of spermatozoa from mice of varying genotype were studied using highspeed cinemicrography. Beat frequency was variable but unimodal in two inbred lines, their F1, and an outbred line. In contrast, beat frequency in spermatozoa from T-complex, balanced lethal stocks (T/t6 and T/t12) tended not to vary between individual males of each genotype up to four hours after collection. The two-hour distribution of beat frequency for T/t12 was, moreover, bimodal, suggesting the possible existence of two subpopulations of spermatozoa.


Changes observed in bone during hibernation using Procion red dye as a matrical marker. Visual examination of the intensity of Procion dye in hamster femurs revealed that dye incorporation decreased during hibernation. Little or no dye was observed after hibernation lasting longer than 50 days. It seems that hibernation may be associated with increased bone resorption and a loss of bone matrix.


Paternal X chromosome expression in extraembryonic membranes of XO mice. The paternally derived allelic form of the X chromosome linked enzyme phosphoglycerate kinase (PGK-1) is expressed in both the fetus and extraembryonic membranes of 9.5 day post coitum XO (XpO) mouse conceptuses. Previous studies on trophectoderm and primitive endoderm derived extraembryonic membranes in XX conceptuses suggest expression of only the maternally derived X chromosome (Xm). Our results suggest that the observed lack of expression of Xp in the trophectoderm and primitive endoderm derivatives of XX embryos is not an intrinsic property of the Xp chromosome.


Effects of divalent cations on in vitro maturation of bovine oocytes. Lowering the external concentrations of both Mg+2 and Ca+2 caused failure of meiotic resumption in vitro of bovine, oocyte-cumulus complexes. Lowering of external Ca+2 levels singly had no effect on either meiotic resumption or completion of the first meiotic division. Lowering of external Mg+2 concentrations alone, although having no effect on meiotic resumption in vitro when Ca+2 was present, did interfere with the completion of the first meiotic division. The result was arrest of oocyte maturation between germinal vesicle breakdown and formation of the first metaphase plate.


Forensic pathology--the hidden specialty: A survey of forensic pathology training available to medical students and residents. It is no wonder that some people refer to forensic pathology as the "hidden specialty." This paper shows evidence that the only exposure given to most medical students is a brief, all-too-explicit demonstration ("horror show") of interesting cases that can only attract the bizarre of mind into the field. It is further pointed out that even residents in pathology receive scanty, if any, formal training in most medical school residency programs, in spite of the fact that two thirds of the medical schools responding have one or more forensic pathologists on their faculty. The obvious answer is a change in the curriculum if this increasingly important subspecialty is to be saved.


A comparative analysis of Cannabis material. Extracts of 100 plant-like or resinous materials were analyzed for CBD, CBC, delta 9-THC, and CBN by GC using two different column packings and by GC-MS. Our independent identification of these cannabinoids confirmed those of other forensic science analysts who used microscopic examination, the Duquenois-Levine color test, and TLC for their analyses of the same samples. The identifications of cannabinoids by forensic acience analysts using TLC were corroborated by GC-MS analysis of hexane extracts of appropriate chromatogram spots.


Incidence of cocaine metabolites in urine specimens from medical examiners' cases. Isobutane CIMS is useful for determining the molecular weight of morphine and its derivatives, as well as for identifying labile acyl substituents on morphine's O-6 position. Furthermore, this technique will provide information relating to the presence or absence of pi-bonding on the C-7 carbon. The spectra of morphine derivatives can be further simplified by employing ethylenediamine as a reagent gas. This approach proves useful for eliciting or confirming molecular weight information from the CI spectrum. In our laboratory extended use of ethylenediamine has been accomplished without any deleterious effect on the mass spectrometer's source or its vacuum system. The utility of isobutane and ethylenediamine CI rests with its ability to supply the analyst with structure elucidation data that may be used to complement more detailed information extractable from either EI or CE spectra. This aspect of mass spectrometry is especially useful when one is dealing with an unknown member of a particular class of organic compounds.


The detection of Y chromosomes in bloodstains--a reevaluation. A method has been described for detecting Y chromosomes in the leukocytes of human bloodstains prepared on a variety of substrates. The factors that influence the proportion of chromosomes exhibiting a Y spot (the Y cell index) in a bloodstain are considered, including the subjective nature of assessment of the Y chromosome fluorescence, the substrate, and the age of bloodstain. In contrast to previous workers no decay in Y cell index with the age of the stain was observed. The results of a blind trial involving stains derived from case work, where from other evidence there was no doubt as to the sex of the donor, are presented. Sixty-five percent of the male bloodstains were correctly identified and no females were wrongly reported as male.


Racial classifications based on blood group protein systems. Two methods employing frequencies of blood group phenotypes are assessed as methods of accurately classifying individuals into racial categories. The data used consist of eleven blood group protein systems from 5043 males distributed into four ethnic categories (white, black, Chicano/Amerindian, and Asian). Both methods work equally well and yield a rate of misclassification of about 22%. Also included is a discussion of two probabilistic calculations relevant to employing blood group protein systems in the context of an identification tool.


Absorption patterns for 33P phosphate into single human and animal hairs. Absorption of phosphate into single human and animal hair shafts has been studied by means of 32P radiotracer techniques. Nondestructive radioassay permitted repetitive experiments on the same hair specimen. Phosphate concentration patterns were shown to be complementary to those for zinc absorption and sometimes to exhibit prominent concentration peaks extending over regions of the shaft 10 to 20 mm in length. Appearance of such peaks was shown to be favored by pH decreasing below 7 but not by migration of phosphate along the hair shaft, which was unimportant.


Separation of standard opiates and their analysis in pharmaceutical and illicit preparations by paired-ion reverse-phase high-pressure liquid chromatography. The separation of standard opiates in a mixture and their analysis in clandestine and pharmaceutical preparations were accomplished by PIC on a micro-Bondapak C18 column. The identification of the opiates was based on two parameters: retention times and the ratios of absorbance peaks recorded at 254 and 280 nm. No prior clean-up procedure of samples was required for analysis by this method. Baseline separation of drug components in clandestine and in pharmaceutical preparations made this method suitable for their quantitation.


Acute epiglottitis (supraglottitis).

Acute epiglottitis (supraglottitis). During an autopsy the clinician, as well as the pathologist, should insist on a culture and adequate examination of the neck. Failure to examine the neck structures in this catastrophic illness may result in an incorrect determination of the cause of death [12,13]. Many autopsy surgeons assume pneumonia is present but later find only microscopic evidence of pulmonary edema. By then the decedent has been buried. Untreated fatalities are seldom reported [3,5,14]. Future reports should include statistics from the medical examiner's or coroner's office in their jurisdiction. This will promote knowledge of the true incidence of acute epiglottitis.


"Paradoxical undressing" in fatal hypothermia. The phenomenon called paradoxical undressing has been described from 33 cases of hypothermia collected from Swedish police reports. The cases were almost evenly distributed with regard to sex, age, and geographical distribution. The cases occurred more frequently in open land although cases from town areas were also found. Most incidents were recorded from November to February at low ambient temperatures, although cases were also reported at temperatures above 0 degree C. Arteriosclerosis and chronic alcoholism were important concomitant illnesses, the latter being frequent in middle-aged men. Epilepsy, diabetes, and pregnancy were present in single cases. Ethanol and other drugs were present in 67% of the males and in 78% of the females, ethanol predominating in men and various psychotropic agents in women. The mean blood ethanol concentration in males was 0.16% and in females, 0.18%. Most frequent findings at necropsy were purple spots or discoloration on the extremities, pulmonary edema, and gastric hemorrhages. It is concluded that paradoxical undressing might be explained by changes in peripheral vasoconstriction in the deeply hypothermic person. It represents the last effort of the victim and is followed almost immediately by unconsciousness and death.


The effect of time of death on extravascular tissue/blood secobarbital concentration ratios in the rat. Extravascular liver/blood and brain/blood ratios were found to be an average of 6% and 1% higher, respectively, in all experiments than total liver/blood and brain/blood ratios. This difference may be informative in establishing true tissue levels. There was a significant time effect (P less than 0.05) with the extravascular liver/blood ratios but not with the extravscular brain/blood ratios. Extravascular liver/blood ratios were slightly higher in phenobarbital-pretreated animals than in non-pretreated animals. Tissue secobarbital levels in pretreated and non-pretreated animals are not different at 1/4 or 1 h, even though pretreated animals received higher doses than non-pretreated animals. Tissue levels are significantly higher (P less than 0.01) in pretreated animals than in non-pretreated animals at 4 h. It is possible that, at this time period, the barbiturate-metabolizing enzymes have become saturated or exhausted.


Reidentification of a major impurity in illicit amphetamine. An impurity previously reported in illicit amphetamine has been found as a by-product om the synthesis of N-formylamphetamine, a reaction precursor to amphetamine by the Leukart synthesis. Its reidentification as di-(1-phenylisopropyl) formamide has been supported by combined spectroscopic analysis of isolated fractions.


Fatal retropleural hematoma from a ruptured abdominal aortic pseudo-aneurysm. This paper has reported a case of sudden unexpected death in a 19-year-old man, shot in the abdomen, who was surgically treated and discharged. He died of massive left retropleural hematoma from a spontaneous rupture of a traumatic pseudo-aneurysm of the abdominal aorta. Diagnosis of the retropleural hemorrhage and the aortic pseudo-aneurysm was possible, though not made antemortem, through chest X-rays and ultrasound studies of the abdomen correlated with the patient's symptoms. Early radiologic signs of the hematoma consisted of obliteration of the aortic silhouette on the anteroposterior view and the left primary sulcus on the lateral film by a convex expanding homogenous density whose medial border blended imperceptibly with the mediastinal shadow. On ultrasound study the pseudo-aneurysm appears as semilucent midline shadow anterior to the vertebral column and related to the aorta, giving the latter an appearance of increased diameter.


A simple method for preparing human sketetal material for forensic examination. A method using dilute sodium hypochlorite for the cleaning of human skeletal material from decomposed bodies is described. With this simple procedure, which requires minimal expenditure of time and money, bones may be cleaned in a manner that produces specimens satisfactory for forensic examination, documentation of injuries, and jury presentation.


Detection of drugs using XAD-2 resin. I:Choice of resin, chromatographic conditions, and recovery studies. Amberlite XAD-2, a nonionic polystyrene divinylbenzene resin, was first used for the analysis of drugs in urine and a number of reports have described the development at optimal conditions for extraction, including type of resin columns, pH conditions, and eluting solvents. XAD-4 and XAD-7 resins were compared to the similarly structured XAD-2 resin and no significant advantage over the XAD-2 resin for drug screening was observed. A quantity of 5 to 6 g of resin was found to have sufficient capacity for the extraction of 200 ml of pentobarbital solution (1 mg/100ml). A column flow rate of approximately 15 ml/min (gravitational flow) was sufficient for analysis and slower rates were not more efficient. A mixture of ethyl acetate and 1,2-dichloroethane (3:2) was found to give best overall recovery (66 to 94%) of drugs, the resulting extracts being reasonably free of interfering substances. A pH value of 8.5 is recommended as optimum for comprehensive analysis of acidic and basic drugs. Recovery studies were conducted on spiked samples to determine drug losses occuring during various steps in the XAD-2 extraction procedure for four acidic (amobarbital, secobarbital, pentobarbital, and phenobarbital) and four basic (morphine, codeine, meperidine, and methadone) drugs. A relatively small amount (0 to 5%) of the drugs was not adsorbed by the resin and amounts varying from 6 to 40% failed to be desorbed by the eluting solvent. Additional losses occurred during the removal and analysis of TLC spots. Recovery of drugs from aqueous solutions analyzed with the XAD-2 resin were compared to recoveries reported in the literature with other XAD-2 resin methods for the extraction of drugs from urine. Recovery of phenobarbital, morphine, and codeine improved by 4 to 23% while recoveries of amobarbital, pentobarbital, secobarbital, methadone, and meperidine were 4 to 28% less efficient when compared to literature data.


Detection of drugs using XAD-2 resin. II:Analysis of liver in medical examiner's cases. Liver tends to concentrate drugs in quantities generally higher than those found in blood or other body compartments. This fact as well as the general availability of liver in postmortem cases makes it an important specimen for comprehensive toxicologic investigation. A scheme for the analysis of liver for drugs with tissue hydrolysis, XAD-2 resin extraction, and TLC has been developed and the parameters affecting recovery have been studied. The hydrolysis of liver specimens at various pH conditions resulted in an improved recovery for morphine by using pH 2 (2N hydrochloric acid). Recoveries of barbiturates, codeine, and meperidine were essentially the same at pH 2 and pH 3. A considerable loss (22 to 55%) was observed for four drugs (pentobarbital, morphine, codeine, and meperidine) as a result of drug binding to the tissue pellets during the process of centrifuging the liver homogenates. This method is recommended as a comprehensive screening procedure for drugs in liver tissue. For quantitative purposes, however, it is necessary to determine a correction factor for all the losses occurring at the various steps of the procedure. This procedure compared favorably with other procedures for liver analysis reported in literature.


Detection of drugs using XAD-2 resin. III:A routine screening procedure for bile. The ability of bile to concentrate drugs and metabolites coupled with its general availability make it suitable for analysis and often the fluid of choice in postmorten cases requiring drug screening. Bile (5 to 10 ml) was diluted with water, sulfuric acid was added, and the mixture was autoclaved. The precipitated bile salts were easily removed by filtration and the filtrate (pH adjusted to 8.0 to 8.5) extracted with XAD-2 resin. Drugs were eluted with a mixture of ethyl acetate/1,2-dichloroethane and analyzed with thin-layer chromatography. Varying the dilution of bile improved the recovery of morphine, codeine, methadone, amobarbital, and phenobarbital. Excessive dilution, however, caused a washing phenomenon and reduced recovery of some drugs, as shown with morphine and codeine. The procedure described is useful for the rapid screening of bile specimens for drugs.


Sudden unexpected death from cardiac concussion (commotio cordis) with unusual legal complications. Sudden and unexpected death may result from cardiac concussion following blunt force trauma to the thorax. Undiagnosed pathologic disease must be carefully evaluated as a possible contributory element. Legal complications may arise from any autopsy. It is recommended that a photograph be taken upon completion of the autopsy. This photograph and adequate records can be used to refute any charges against the pathologist or assistants for the poor condition of a body after its release.


Sudden death and sepsis after splenectomy. This paper has reported two deaths occurring in young adults who had undergone splenectomy for trauma several years before developing pneumococcal sepsis. Tissues at autopsy demonstrated a disproportionate autolysis for the postmortem interval. One victim also showed diffuse serosal hemorrhages, presumably as a result of disseminated intravascular coagulation. Both showed evidence of residual splenic implants but such implants clearly did not provide protection against sepsis. The mechanisms whereby the spleen protects from sepsis appear to be that of a mechanical filter and an immunological organ producing antibody or antibody-like substance. How much splenic tissue and what relationships to the system circulation are necessary to provide protection remain undiscovered.