Search

Hyperspectral imaging (HSI) has become a crucial innovation in forensic science, particularly for analysing bodily fluids. This advanced technology captures both spectral and spatial data across a wide spectrum of wavelengths, offering comprehensive insights into the composition and distribution of bodily fluids found at crime scenes. In this review, we delve into the forensic applications of HSI, emphasizing its role in detecting, identifying, and distinguishing various bodily fluids such as blood, saliva, urine, vaginal fluid, semen, and menstrual blood. We examine the benefits of HSI compared to traditional methods, noting its non-destructive approach, high sensitivity, and capability to differentiate fluids even in complex mixtures. Additionally, we discuss recent advancements in HSI technology and their potential to enhance forensic investigations. This review highlights the importance of HSI as a valuable tool in forensic science, opening new pathways for improving the accuracy and efficiency of crime scene analyses. • Hyperspectral imaging enhances non-destructive detection of fluids at crime scenes. • Ability to precisely identify and differentiate complex fluid mixtures. • Various detection methods for blood, saliva, urine, vaginal fluid, & semen. • Compares hyperspectral imaging with traditional forensic methods. • Adaptability provides advantages over conventional techniques. [ABSTRACT FROM AUTHOR]

Vibrational spectroscopy offers a unique opportunity to investigate the composition of unknown substances on a molecular basis. The spectroscopy of molecular vibrations using mid-infrared or Raman techniques has been applied to samples of body fluids. This review presents some applications related to body fluids published in the period 2005–2015. [ABSTRACT FROM PUBLISHER]

Abstract: Trace elements play an important role in the human central nervous system. Significant variations of the concentration of trace elements in body fluids may occur in neurodegenerative diseases. In the present work an investigation of the elemental composition of the serum, and the cerebrospinal fluid in amyotrophic lateral sclerosis patients and a control group was performed. For the analysis of the body fluids Total reflection X-ray Fluorescence (TXRF) spectrometry was used. The samples were taken during routine diagnostic procedures. Na, Mg, Cl, K, Ca, Cu, Zn, and Br were determined in both fluids. In order to validate the results of analysis a serum standard reference material was measured. A t-test was applied to check if the mean concentrations of the elements are different for ALS and the control group. For the serum samples higher values for Br were found in the ALS group, for the cerebrospinal fluid lower values of Na, Mg and Zn as well as higher Ca values were found in the ALS group compared to the control group. [Copyright &y& Elsevier]

The coelomic fluid (CF) of Asterias forbesi was examined for the presence of complement-like activity. Cell-free CF contains hemolytic activity for unsensitized rabbit erythrocytes (E) but not unsensitized or antibody-coated sheep E. Target E sensitivity is not dependent upon phylogenetic proximity of erythrocyte species to Asterias. The activity is optimally assayed at 25 degrees C, has a sigmoidal dose-response curve, and is heat-sensitive (56 degrees C). The activity is divalent cation dependent, although Ca++ and Mg++ are not required. Nonlytic CF does not contain an inhibitor; rather, combining nonlytic and lytic CFs results in an increase in hemolysis. Activity can be consumed by treatment with zymosan, pronase, trypsin, and PMSF. Enzymatic desialidation converts sheep E into activators of the hemolytic pathway. No phospholipase activity is present in CF. Biological function is demonstrated by bactericidal activity against Vibrio tubiashii by lytic CF. These results suggest that a primitive complement system is operating in Asterias.

The inhibin content and aromatase inhibitor activity (AIA) of 72 follicular fluids (FF) obtained from 42 women undergoing in vitro fertilization (IVF) and embryo transfer (ET) were studied as a function of IVF ET outcome. Inhibin levels were determined by bioassay (BA) and RIA; AIA was measured by BA. The inhibin content of follicles characterized as immature by their estradiol (E2) levels and E2/progesterone (P) ratios was significantly lower (P less than 0.05) than that of mature follicles (i.e. leading to pregnancy). The mean AIA for mature follicles were significantly lower than AIA in groups where pregnancy was not obtained. AIA for follicles from which a pregnancy was obtained for each ET was also significantly lower than that in FF characterized as immature of hypermature. The highest E2/AIA and inhibin BA/AIA ratios were associated with the highest incidence of successful IVF ET outcome. No correlation was found between AIA and inhibin, on the one hand, and E2, delta 4-androstenedione, E2/P, and PRL, on the other. However, a positive correlation was found between inhibin (RIA and BA) and P, reflecting the production of inhibin by granulosa cells during luteinization. These studies allowed us to conclude that FF inhibin levels do not differ according to IVF ET outcome, but are an index of follicular maturation. AIA not only constitutes an index of follicular maturation and granulosa cell luteinization, but is of predictive value for IVF ET outcome as E2/AIA and inhibin BA/AIA ratios.

The HER2/neu proto-oncogene encodes a receptor that belong to the tyrosine-specific protein kinase family. Amplification of the HER2 gene in patients with breast and ovarian cancer has been shown to predict poorer survival rates. In order to understand the role of HER2 in malignant and normal cells, it is necessary to devise assays that can quantitate expression levels of the HER2 gene product (p185HER2) in production samples, biopsy specimens and biological fluids. We have developed a simple, quantitative ELISA that uses two monoclonal antibodies directed against the extracellular domain of the HER2 gene product, p185HER2 (HER2 ECD). The assay has a detection range of 0.25-120 ng/ml, is precise and sensitive. The ability of this assay to detect biologically active rHER2 ECD is demonstrated by its correlation to a growth inhibitory bioassay (r = 0.92). The sandwich ELISA can also accurately quantitate rHER2 ECD in mouse and monkey serum. This assay should be useful for quantitating low levels of circulating rHER2 ECD in animals in which rHER2 ECD is being used as antigen for immunotherapy and in patients which 'shed' receptor.

Concentrations of alpha 1 acid glycoprotein, albumin, transferrin, haptoglobin, immunoglobulins G, A, M and apolipoprotein B were measured in serum and suction blister fluid from a group of individuals presenting a biologically proven inflammatory syndrome, and from a control group. Protein values in suction blister fluid did not change from the 2nd to the 3rd h after the beginning of blister formation. The ratio of the concentration of proteins in blister fluid and serum did not differ significantly between the groups. However, a 25% decrease in blister fluid albumin and a 100% increase in blister fluid alpha 1 acid glycoprotein, recorded in the inflammatory group, were worth noting, since they possibly influence the tissular distribution of some protein-binding drugs. Finally, an inverse relationship was established between the blister fluid/serum concentration ratio and the respective molar mass of each protein.

The polar metabolites of polycyclic aromatic compounds (PACs) are of significant oncological interest. In contrast to parent PACs, isomer selective detection of polar PACs by laser excited Shpol'skii spectrometry (LESS) is severely limited by excessively broadened spectra and rapid photodegradation in n-alkane solvents. To minimize these limitations a 10-min derivatization procedure was developed to produce corresponding methoxy compounds, that exhibit the Shpol'skii effect. By use of the extraction and permethylation procedure, the selective detection of a mixture of hydroxy-benz[a]anthracene isomers spiked at the low picogram level into urine and blood matrices was achieved. A detection limit for 1-hydroxybenz[a]anthracene was 0.6 part per trillion, or 12 fg (0.05 fmol) on a 20-microL sample.

A colorimetric method for the screening of cyanide in biological specimens is presented. Cyanide in a 200-microL sample is diffused in a Conway 3 compartment dish into the center well where it catalyzes the reaction between p-nitrobenzaldehyde and o-dinitrobenzene. The resulting blue-violet color intensity is proportional to cyanide concentration. The reaction is specific for cyanide, and concentrations as low as 0.05 mg/L can readily be detected.

Positive-ion electron impact (PIEI), positive-ion chemical ionization (PICI) and negative-ion chemical ionization (NICI) mass spectra of 9 carbamate pesticides are presented. In the PIEI mode, the spectra showed small molecular peaks, intense or base peaks due to M - CH3NHCO + H and peaks at m/z 58 due to CH3NHCO. In the PICI mode, peaks due to M + H, M + C2H5, M - CH3NHCO + 2H, CH3NHCO(m/z 58) and M-28 appeared. The cations at m/z 58 found in both PIEI and PICI modes seem very useful for screening of a carbamate. In the NICI mode, the spectra showed peaks due to M - CH3NHCO and characteristic anions appearing at mass numbers higher than molecular ones, which were probably due to dimerization of [M - CH3NHCO]-followed by hydrogen attachment. Carbamates, which had been added to urine, plasma, whole blood, the liver, kidney and brain, could be rapidly isolated by use of Sep-Pak C18 cartridges with chloroform as an elution solvent. They could be detected by wide-bore capillary gas chromatography with a SPB-5 column, with satisfactory separation from impurities in their underivatized forms.

A sensitive enzyme-linked immunosorbent assay (ELISA) for detection of ABO blood substances in human body fluids was devised. The ELISA plates coated with purified human anti-A or anti-B serum effectively captured the blood substances, and these were then analysed by the combination of rabbit anti-A and goat anti-B. This capture ELISA could differentiate the type AB specimen from a mixture of the type A and the type B specimens, and the method was applied to rape cases to make the ABO typing of the criminal.

This prospective, randomized, single-blind study of 121 healthy children aged 2 to 12 yr investigated the effect of clear fluids on gastric contents. Gastric fluid volume and pH were measured immediately following the induction of general anaesthesia and were not significantly affected by the ingestion of unlimited clear fluids up to three hours preoperatively. After a prolonged fast (mean fast 14 hr), gastric fluid volume was 0.39 +/- 0.37 ml.kg-1 and gastric pH was 1.7 +/- 0.4; after unlimited clear fluids (203 +/- 109 ml) up to three hours before surgery gastric fluid volume was 0.34 +/- 0.28 ml.kg-1 and gastric pH was 1.8 +/- 0.7 (mean +/- SD). Gastric fluid volume (ml.kg-1) increased in both the control and study groups as age increased, P less than 0.005. It is concluded that drinking clear fluid up to three hours before scheduled surgery does not have a measurable effect on gastric volume and pH of healthy children of ages 2 to 12 yr.

Measurements of specific antibody titres in uterine fluid of mice immunized by different routes indicated that two immunizations in the pelvic presacral space using aluminium hydroxide as adjuvant was a simple and effective way to elicit a significant IgA and IgG response. Higher IgA and IgG titres were produced in uterine fluid by subcutaneous immunization with antigen in Freund's complete adjuvant followed by intravaginal boosting without adjuvant, but this immunization involved both a toxic adjuvant and repeated applications of large doses of antigen in the vagina. Intragastric immunization produced an IgA response in the uterus but no IgG. Local intravaginal priming and boosting with large doses of antigen without adjuvant produced an IgA response in uterine fluid, but was less effective for IgG and was inefficient in terms of time and the amount of antigen used. Hysterectomy reduced the concentration of specific IgA in vaginal fluid of immunized mice to no more than 5% of normal, indicating that most of the IgA in vaginal fluid originates in the uterus. In contrast, IgG titres were not significantly different in hysterectomized and intact mice. IgA titres in vaginal fluid were at least partly restored to normal levels in sham-hysterectomized mice.

Immune responses in the uterus are depressed when concentrations of progesterone in the blood are elevated. We tested whether this action of progesterone involves induction of secretion of immunosuppressive molecules into the uterine lumen. Uterine fluid from ovariectomized ewes treated with progesterone for 60 days inhibited [3H]thymidine (TdR) incorporation by phytohemagglutinin-stimulated lymphocytes more than uterine flushings from vehicle-treated ewes. Uterine fluid from progesterone-treated ewes also inhibited TdR uptake by other mitogen-stimulated lymphocytes and mixed lymphocyte cultures, but had no effect on the growth of three nonlymphocyte cell lines. A group of ewes immunized with ovalbumin mixed with uterine fluid from progesterone-treated ewes produced an antibody titer that was lower than the titer for ewes immunized with ovalbumin without uterine fluid. Analysis of the physical properties of the active molecules in uterine fluid from progesterone-treated ewes indicated that activity could be separated into two fractions: a large mol wt (Mr; greater than 5000) fraction that was sensitive to pronase and a low Mr (less than 1000) fraction that was more resistant to pronase. There was more inhibitory activity in the high mol wt fraction. Further analysis of this fraction indicated that most activity was of basic isoelectric point (pI greater than 8.2). The most active fraction of basic material eluted at the void volume of a Sepharose CL-6B column (Mr, 4 x 10(6)). In conclusion, progesterone caused accumulation of factors in the uterine lumen that inhibited lymphocyte function in vitro and antibody formation in vivo. These molecules may play an important role in regulating immune responses in the uterus during pregnancy.

Plasma D2O-accumulation profiles (qualitative indices of fluid-absorption rates) were determined in eight subjects after ingestion of 275 mL of five D2O-labeled beverages: a water placebo (W), 6% maltodextrin (6% M), and three solutions containing a 6%, 8%, and 10% glucose-fructose mix (6% GF, 8% GF, and 10% GF). Except for W all beverages contained 20 mmol sodium/L and 3 mmol potassium/L. No differences in plasma D2O accumulation were found. Plasma glucose increased at 20 and 30 min after ingestion of the carbohydrate drinks and returned to baseline (6% GF and 6% M) or below (8% GF and 10% GF) by 60 min. Insulin responded similarly and, except for a slightly lower value at 30 min for 6% GF, no differences were detected. It appears that fluids in drinks containing less than or equal to 8-10% carbohydrate (simple sugars or maltodextrins) are made available for dilution in body fluids at similar rates and should be similar in replenishing body fluids lost in sweat during exercise.

In order to investigate the role of renin angiotensin in the epididymis, angiotensin-converting enzyme (ACE) activity and angiotensin I (AI) and angiotensin II (AII) concentrations were measured in the male reproductive tract and blood serum of the rat. High ACE activity was detected in the rat epididymis, with a major part of the activity being associated with epididymal spermatozoa. When spermatozoa were prevented from entering the epididymis by efferent duct ligation, the ACE activity in the epididymis was greatly reduced. The epithelial cells lining the epididymal duct were also shown to possess ACE activity which was dependent upon circulating androgens. Treatment of male rats with captopril at a single oral dose (20 mg/kg) significantly inhibited the ACE activity in the blood serum but had no effect on the activity of the epididymal fluid. The intraluminal ACE was protected from the circulating captopril by the blood-epididymis barrier. Long-term treatment with captopril (20 mg/kg per day, 8 weeks), however, caused an increase in blood serum ACE activity but was without effect on intraluminal ACE. The fertility and fecundity of male rats after treatment were apparently normal. The concentrations of AI and AII were high in the epididymal plasma and epididymal cell when compared with the respective concentrations in blood serum. The intraluminal AII concentration found (13 nmol/l) was close to the threshold concentrations that stimulate anion (and fluid) secretion in cultured epididymal epithelium in vitro. The high intraluminal AII concentration could not have been derived from the testicular fluid or spermatozoa since the rete testis fluid and sperm contained little AII.(ABSTRACT TRUNCATED AT 250 WORDS)

Nuclear magnetic resonance spectroscopy is the epitome of the high-technology, expensive diagnostic method. Extrapolation from a limited number of patient examinations and from experiments in animal models predicts a bright future for the method. However, several barriers block widespread clinical application in the near future; technical difficulties still exist but they seem to be resolvable in due course. A more serious problem is the absence of an adequate database from which to interpret the vast array of information produced by nuclear magnetic resonance. The necessary understanding of the pathologic biochemistry of disease will be frustratingly slow to appear as will the routine clinical use of magnetic resonance spectroscopy. The critical need for improved diagnostic methods will stimulate experimentation to resolve these problems.

Specific and sensitive reverse-phase HPLC assays of buprenorphine and its metabolite in biological fluids were developed with sensitivities of 2-6 ng ml-1 using fluorimetric detection. Pharmacokinetics were monitored on acute bolus administration of buprenorphine in 6 dogs within the 0.7-2.6 mg kg-1 dose range. Toxicity was circumvented when terminal plasma concentrations were increased by infusing 3.7-4.8 mg kg-1 doses of buprenorphine over 3 h in six studies in 6 dogs. The terminal rate constants of the IV infusion studies from the triexponential fits of plasma concentration-time data averaged 41.6 +/- 7.5 h with an averaged total body clearance of 191 +/- 19 ml min-1. This terminal rate constant was in contrast to the less than 100 min half-life of the second exponential fitting of the less lipophilic morphine, naloxone, and naltrexone. The apparent volumes of distribution of buprenorphine, referenced to the total plasma concentration, were 33 +/- 61 (Vc, central compartment volume) and 663 +/- 891 (Vd, total body volume), indicative of a highly bound, sequestered or lipophilic drug. Unchanged buprenorphine was insignificantly renally (less than 0.2 per cent of the dose) and biliary (less than 0.6 per cent) excreted. The major route of buprenorphine disposition was by hepatic conjugation to glucuronide which was eliminated into the bile (about 92 per cent) with only small amounts appearing in urine (less than 1 per cent as metabolite). Minor metabolites excreted in the bile accounted for about 3 per cent of the administered dose. Direct IV administration of the metabolite, buprenorphine glucuronide, gave a terminal half-life of 6 h and more than 90 per cent of the systemically circulating metabolite was excreted in bile; only 10 per cent in urine. The oral bioavailability, estimated from the areas under the buprenorphine plasma concentration-time curve following IV and oral administration of buprenorphine in the dogs, was 3-6 per cent. There were no apparent correlations of the buprenorphine time course with cardiovascular parameters such as heart rate, ECG, and blood pressure. Miotic effect was significant. Respiratory depression was observed during the first 4 h after IV bolus injection, but not during the infusion studies.

The use of stable isotopically labeled steroids combined with gas chromatography/mass spectrometry (GC/MS) has found a broad application in pharmacologic studies. Initially, stable isotopically labeled steroids served as the ideal analytic internal standard for GC/MS analysis; however, their in vivo use has expanded and has proven to be a powerful pharmacokinetic tool. We have successfully used stable isotope methodology to study the pharmacokinetic/bioavailability of androgens. The primary advantage of the technique is that endogenous and exogenous steroids with the same basic structure can be differentiated by using stable isotopically labeled analogs. The method was used to examine the pharmacokinetics of testosterone and testosterone propionate, and to clarify the influence of endogenous testosterone. Another advantage of the isotope methods is that steroidal drugs can be administered concomitantly in two formulations (e.g., solution and solid dosage). A single set of blood samples serves to describe the time course of the formulations being compared. This stable isotope coadministration technique was used to estimate the relative bioavailability of 17 alpha-methyltestosterone.

Hypomagnesemia is a common clinical finding in hospitalized patients and can cause hypocalcemia, cardiac arrhythmias, muscular weakness, and hypokalemia. Hypomagnesemia usually implies cellular magnesium (Mg) depletion, but stress and some clinical conditions which raise serum catecholamine concentrations may lower serum Mg (sMg) concentrations. To help investigate the mechanism and degree of the effect of catecholamines on sMg concentration, we gave intravenous epinephrine (0.1 microgram/kg/min) to 12 normal volunteers for 2 hours. The sMg concentration fell from 1.86 +/- 0.04 mg/dl to 1.63 +/- 0.05 mg/dl (mean +/- SEM, p less than 0.01). Pre-infusion intracellular free Mg (Mg++) in red blood cells (RBC) as measured by nuclear magnetic resonance spectrophotometry (NMR) was 171 +/- 7.6 microM and did not differ significantly from post-infusion RBC Mg++, 186 +/- 12.6 microM. Total blood mononuclear cell Mg content and urine Mg excretion also did not change. These data suggest that epinephrine has a small but significant effect on the lowering of sMg concentrations. Endogenous catecholamine release during stress or acute illness may therefore contribute to the hypomagnesemia seen in acutely ill patients. Our data also suggest that hypomagnesemia seen under conditions of acute stress may not always imply depleted tissue Mg stores. As no absolute change in cellular Mg or in urinary Mg excretion was demonstrated, acute intracellular shifts of Mg into blood cells and/or urinary Mg losses may not account for the hypomagnesemia. The prevalence and clinical consequences of stress hypomagnesemia require further investigation.

A paired-ion high-performance liquid chromatographic method was developed to measure concentrations of 2',3'-dideoxyinosine (ddI) in human plasma, urine and cerebrospinal fluid. Samples were prepared using a solid-phase extraction technique which allows for a five-fold concentration of the drug. 2'-Deoxyguanosine was added as an internal standard prior to the extraction. Recoveries for 2'-deoxyguanosine and ddI were 80 +/- 15 and 85 +/- 10%, respectively. Extracted samples were then injected onto a C18 column and eluted isocratically with a mobile phase containing 0.1% of the ion-pairing reagent, heptafluorobutyric acid, and 5% acetonitrile. The retention time was 7.4 min for 2'-deoxyguanosine and 8.4 min for ddI. The lower limit of detection for ddI is 0.1 microM. Using this technique the acid lability of ddI was demonstrated and the plasma concentration versus time profile from a patient receiving the drug was examined.

A two-day technical workshop was convened November 10-11, 1986, to discuss analytical approaches for determining trace amounts of cotinine in human body fluids resulting from passive exposure to environmental tobacco smoke (ETS). The workshop, jointly sponsored by the U.S. Environmental Protection Agency and Centers for Disease Control, was attended by scientists with expertise in cotinine analytical methodology and/or conduct of human monitoring studies related to ETS. The workshop format included technical presentations, separate panel discussions on chromatography and immunoassay analytical approaches, and group discussions related to the quality assurance/quality control aspects of future monitoring programs. This report presents a consensus of opinion on general issues before the workshop panel participants and also a detailed comparison of several analytical approaches being used by the various represented laboratories. The salient features of the chromatography and immunoassay analytical methods are discussed separately.

There is a cytotoxic activity in blastocele fluid that kills embryonal carcinoma cells with trophectodermal potential but spares those with embryonic potential. This activity is present when programmed cell death occurs in the inner cell mass (ICM), and the ICM loses its trophectodermal potential. Because of the paucity of blastocele fluid, cystic embryoid bodies of embryonal carcinoma C44 were examined ultrastructurally and in tissue culture to determine if they corresponded to late blastocysts and if their fluid corresponded to blastocele fluid. No trophectoderm was demonstrated in the embryoid bodies, but embryonal carcinoma and endoderm were present, leading to the conclusion that the embryonal carcinoma corresponded to late ICM that had expressed endodermal potential. As a result the cyst fluid might have contained the toxic activity of blastocele fluid. The cyst fluid of C44 embryoid bodies did contain a soluble, low-molecular-weight, cytotoxic activity that preferentially killed embryonal carcinoma cells with trophectodermal potential while sparing those with embryonic potential. Enough of this fluid was available to determine the chemical nature of this toxic activity.

A common technique to sample airway mucosal 'surface' liquids is with absorbing discs of filter paper. The present study examined the efficacy of this technique by analysing tracheal liquids of control and capsaicin (0.1 nmol)-exposed guinea-pig airways. Mucosal fluids, obtained by topically applied discs or by a specific lavage procedure, and tracheal tissue were sampled. The animals had received FITC-dextran (MW 70 kDa) intravenously and this specific plasma tracer was analysed in the sampled material. Under control conditions significantly more FITC-dextran was found in the discs than in the tracheal lavage fluids (P less than 0.001) despite the fact that the lavaged mucosal surface was much larger than that covered by the discs. Capsaicin significantly increased the content of FITC-dextran in all fluids sampled as well as in the airway tissue. In all cases concentrations of FITC-dextran in the disc fluids did not differ much from that in the tissue samples. These data suggest that absorbing discs severely disturb the epithelial-barrier function and sample subepithelial fluid and solutes including macromolecules. As demonstrated in this study by the elevated content of a plasma tracer molecule an inflammatory process may, nevertheless, be traced in the mixture of surface and tissue fluids that is sampled by the discs.

Antibodies were produced in sheep against a new anti-breast cancer drug 4-hydroxyandrostenedione (4-OHA) using two hapten-ovalbumin conjugates. One of these conjugates (4-hydroxytestosterone-ovalbumin) produced an antiserum suitable for the development of a radioimmunoassay that would allow direct measurement of 4-OHA in plasma at concentrations down to 82 pmol/l, with adequate accuracy, precision and scope for further sensitivity. Although this assay would measure 4-hydroxytestosterone (4-OHT) in addition to 4-OHA, the present data suggest that the magnitude of any interference from endogenous steroids and those derived from 4-OHA could only be minimal. A comparison of solvent-extracted and unextracted samples showed that only unconjugated drug was analysed by this radioimmunoassay. A study of plasma protein binding of 4-OHA showed that at therapeutic concentrations, between 13.5 and 16.5% of plasma 4-OHA was not bound to proteins. This assay system could be a useful adjunct to the future development of 4-OHA as an anti-cancer drug.

In the present paper we analyze the importance of gastro-oesophageal reflux in 20 patients with Barrett's oesophagus and in 20 patients with esophagitis without Barrett's mucosa; ten of this last group had mild esophagitis and ten severe inflammatory changes. In all the cases the oesophageal pH was measured during 24 hours; the results showed that although the reflux was more important in the group of patients with Barrett's esophagus than in the whole group of patients with esophagitis without Barrett's esophagus, figures were similar in the group with severe oesophagitis and the group with Barrett's oesophagus. We conclude that the pathogenesis of Barrett's esophagus includes factors other than gastroesophageal reflux.

An ion chromatographic method was used to simultaneously determine nitrate and nitrite ions in biological samples. Ultrafiltration was used to produce a protein-free filtrate. Chloride interferences were eliminated by precipitation as the silver salt. Detection limits and average recoveries were 0.5 mg/L and 102% for nitrate and 0.2 mg/L and 78% for nitrite, respectively. Nitrate concentration was 2.1 +/- 1.8 mg/L and 4.9 +/- 0.8 mg/L in serum and ocular fluid of healthy cattle, respectively; nitrite was not detected. A severe case of nitrate poisoning in cattle was described and used to study the concentrations of nitrate and nitrite in samples obtained under natural conditions. Nitrate concentration of acutely poisoned cattle was 35% lower in ocular fluid at 158.1 +/- 51.4 mg/L, than in serum at 256.3 +/- 113.4 mg/L. Nitrite was not detected, because of the long processing time (greater than 3 hours) required for samples obtained in the field. A gradual decrease in ocular fluid nitrate of 29.4% at 24 hours, 25.9% at 36 hours, 51.6% at 48 hours, and 73.2% at 60 hours was observed; however, concentrations remained diagnostically significant (73.2 mg/L) 60 hours after death. Twenty-four hours after poisoning, the serum nitrate concentration of severely ill (52.7 +/- 51.9 mg/L) and moderately affected (12.4 +/- 5.7 mg/L) cattle that survived was indicative of the severity of clinical signs previously observed. Nitrate in serum and ocular fluid was stable in samples stored for 24 hours at 23 C, 1 week at 4 C, and 1 month at -20 C.

Previous in vivo studies have demonstrated that alpha, beta, and gamma ATP chemical shifts measured by 31P NMR spectroscopy can be used to determine intracellular magnesium in erythrocytes, but up to now such results have not been confirmed in striated muscle in vivo. We report beta ATP chemical-shift in vivo measurements revealing the depletion of free intracellular magnesium in striated muscle of rats fed a magnesium deficient diet.

The high-performance liquid chromatographic assay described permitted a simple, rapid, sensitive, selective and precise quantitative determination of eugenol in body fluids (serum, urine and bile) without derivatization. Amounts in the range 0.02-100 micrograms of eugenol per millilitre of body fluid were determined with intra-assay coefficients of variation below 4% (3.72-1.13%). The short analysis time for each sample and the selectivity even at low concentrations made this assay suitable for pharmacokinetic studies. Eugenol undergoes a pronounced first-pass effect; in serum, unconjugated eugenol was not detected after an oral dose of 150 mg. The kinetics of eugenol conjugates were measured. More than 80% of the dose was excreted within 6 h after oral administration.

A total of 390 body cavity fluids were analyzed by both cytopathologic examination and flow cytometric DNA analysis. The two methods gave compatible results in analyses of 304 fluids (78%). In 24 patients, cytopathologic studies found the specimens to contain malignant cells, but the DNA content was diploid. This illustrates an area where flow cytometric studies do not extend tumor detection. In 56 fluids from 48 patients, cytologic methods revealed no malignant cells but flow cytometry distinguished aneuploid cell populations; additional clinical information allowed the identification of malignant tumors in 24 (50%) of these patients. Because flow cytometry was able to detect aneuploidy in cases where conventional cytologic examination could not detect malignant cells, the number of patients with tumors detected was increased by 39% beyond those detected by cytologic methods alone in this series.

Bovine oviductal fluid (OF) was collected and analyzed throughout the estrous cycle, and the capacity of the protein and lipoprotein components to support cholesterol efflux from bovine sperm was evaluated. Blood was collected and assayed for progesterone (P4) to monitor the estrous cycle. Protein and lipoprotein separation was achieved by density gradient centrifugation. Two major bands were identified. The first (1.056 less than delta 20 less than 1.140 g/ml) corresponded to bovine and rabbit plasma high-density lipoprotein (HDL) based on distribution in the density gradient and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The second band (1.235 less than delta 20 less than 1.243 g/ml) consisted predominantly of oviductal fluid albumin (OFA). Oviductal fluid protein concentration increased as serum P4 decreased around the time of estrus. Mean OF protein concentration was 21.3 mg/ml when serum P4 was lower than 0.5 ng/ml and 6.9 mg/ml when serum P4 was greater than 0.5 ng/ml. An inverse log relationship was found between HDL protein concentration and serum P4. Unesterified cholesterol (UC), cholesteryl ester, and phospholipid (PL) content of HDL for HDL protein concentrations of 3-56.1 micrograms/ml were 1.35-46.2 micrograms/ml, 1.91-44.48 micrograms/ml, and 1.69-59.8 micrograms/ml, respectively. Phosphatidylcholine and -ethanolamine were the major PLs present in the HDL fraction and their molar ratio (4:1 mol/mol) was relatively constant through the estrous cycle. The OFA fraction of the same samples accounted for more than 90% of total protein and for most of the variation in OF protein. To determine the ability of OF components to serve as sperm cholesterol acceptors, OF samples were incubated 1:1 (v/v) with and without 4 X 10(8) bovine sperm in 1.0 ml of modified Tyrode's solution and OF for 2 hr at 39 degrees C. After incubation, HDL and OFA fractions were isolated and analyzed for changes in protein and lipid content. After OF, samples were incubated with sperm, an increase in UC was found in the HDL fractions. UC in HDL increased by 12.1 +/- 1.0 micrograms/ml (means +/- SE) when serum P4 was less than or equal to 0.5 ng/ml. For samples corresponding to higher serum P4, the increase in UC was 3.60 +/- 0.89 micrograms/ml. Values for UC in HDL were corrected for the contribution of UC from OFA of OF samples. Cholesterol efflux from sperm has been implicated in the process of sperm capacitation. These results indicate that HDL from OF is elevated during the follicular phase of the estrous cycle and can serve as an acceptor for bovine sperm cholesterol.

Closure of full-thickness open wounds in loose-skinned animals is accomplished by wound contraction. Fibroblast-populated collagen lattice (FPCL) contraction is an in vitro model for studying wound contraction. Fibroblasts suspended in a collagen matrix reorient the surrounding collagen fibers, resulting in a reduction in the size of the FPCL. The organization of collagen fibers by fibroblast-generated forces produces lattice contraction. An open wound in a rat begins to show contraction by 3 days, and its size will be reduced by 50% at 7 days. Fluid from 3- and 7-day-old rat wounds was examined for its ability to affect in vitro lattice contraction. Wound fluid was found to inhibit lattice contraction. The fractions which inhibit lattice contraction had molecular weights ranging between 10,000 and 20,000, as revealed by molecular sieve chromatography, and a high positive charge, as demonstrated by ion exchange chromatography. The factor(s) was only slightly affected by added indomethacin in the FPCL contraction model. This suggests a mechanism independent of the generation of prostaglandins. The factor(s) was tested in an ATP-induced model of fibroblast contraction where it was shown to be ineffective at altering cell contraction. The factor(s) did, however, prevent cell spreading and elongation on glass surfaces. Wound fluid has a factor(s) which hinders fibroblast spreading and elongation and which inhibits FPCL contraction.

Reversed-phase high-performance liquid chromatography (RP-HPLC) is a powerful method for assaying physiological amino acid concentrations in biological fluids. Four pre-column derivatization methods, with o-phthaldialdehyde (OPA), 9-fluorenylmethyl chloroformate (FMOC-Cl), phenyl isothiocyanate (PITC) and 1-dimethylaminonaphthalene-5-sulphonyl chloride (dansyl-Cl), were assessed with respect to their applicability in biological research. The methods permit the measurement of 21-26 major amino acids in 13-40 min. The superior sensitivity favours the use of OPA, FMOC-Cl and dansyl-Cl techniques. Because of instability of the OPA adducts, automated on-line derivatization is required when using this method in general practice. Application of the PITC method, although less sensitive, is useful in clinical chemistry, where sample availability is rarely a problem. Cystine determination is not feasible when using OPA or FMOC-Cl and with PITC the reproducibility and linearity are poor, whereas the dansyl-Cl method allows reliable quantitation. The four methods are currently used to perform ca. 8000 OPA and 5000-6000 FMOC-Cl, PITC and dansyl-Cl analyses of biological samples per year. The results obtained with the RP-HPLC methods compare favourably with those derived from conventional ion-exchange amino acid analyses. When the guard column is regularly changed after 120 analyses, the separation remains satisfactory for at least 700 OPA, 800 FMOC-Cl, 150 PITC and 500 dansyl-Cl analyses. Careful control of factors and limitations inherent in the various methodologies is a prerequesite for proper identification and appropriate quantitation.

Extensive subcutaneous and intermuscular calcium laden fluid collections (milk of calcium) were found in two young girls with severe dermatomyositis. Sonographic examination clearly showed the nature and extent of the collections. Knowledge of this new feature of dermatomyositis should help avoid confusing these fluid collections with soft tissue infection and deep abscess in these often steroid dependent children.

Placental protein 19 (PP19) is one of the new placental tissue proteins identified in extracts from human term placenta by Bohn and Winkler. We measured the PP19 concentration in body fluids and placental tissue by radioimmunoassay; the minimum detectable dose of standard was 1.5 ng/ml. Although ethylene diamine tetraacetic acid (EDTA-2K) inhibited the immunoreaction between PP19 (225/242) and anti-PP19 antibody (632 ZA), the PP19 concentration did not differ between serum and heparin and sodium citrate plasmas. The serum PP19 concentration was increased by hemolysis. In blood cell fractions separated by the Ficoll-Paque/Macrodex method, polymorphonuclear leukocyte fraction contained the highest PP19 concentration. The circulating serum PP19 concentration was 4.5 +/- 1.1 ng/ml (mean +/- standard deviation) in the proliferative phase (n = 8) and 5.1 +/- 1.6 ng/ml in the secretory phase (n = 7) for nonpregnant women, and 4.6 +/- 2.2 ng/ml from men (n = 12). Seminal plasma (n = 8) contained 212.2 +/- 99.7 ng/ml. The maternal serum PP19 concentration in 291 normal pregnancies increased from 6.2 ng/ml (median) at 6-7 weeks of gestation to 34.1 ng/ml at 38-39 weeks. The mean PP19 concentration was higher in amniotic fluid and retroplacental blood, but lower in umbilical cord blood than that in circulating maternal serum. In hydatidiform mole, vesicular fluid contained high PP19 concentration (1154.6 +/- 659.5 ng/ml), although these maternal serum concentration was not statistically higher than normal range. The chorionic villous trophoblast contained more PP19 than decidua, chorion, and amnion. These results suggest that PP19 has an extraplacental source, even though the chorionic villous trophoblast may be the main source throughout pregnancy.

Reactive pyrrolic metabolites formed when pyrrolizidine alkaloids are given to rats can alkylate soluble and tissue-bound thiol groups. Pyrrolic thioethers thus formed are relatively stable, and may persist in tissues for long periods. A simple procedure has been developed for recovering the nucleus of the pyrrolic metabolite from such S-binding, whether in solution or attached to solid tissues, in an easily identifiable form. The thioether bond was broken by silver nitrate and the pyrrolic moiety allowed to react with ethanol or methanol to form an ethoxy or methoxy derivative. The chemical basis of the procedure was established by model experiments on a preparative scale, but for small scale recovery from tissues, pyrrolic ethers were extracted and identified by TLC, HPLC, capillary GC and mass spectrometry. Because the pyrrolic derivatives thus formed were easily recognised and unrelated to any physiological compound, the recovery method described, especially when applied to blood samples, provided a way to monitor animals for previous exposure to toxic pyrrolizidine alkaloids.

The 300 MHz (7 T) water proton resonances of suspensions of red blood cells containing paramagnetic deoxyhemoglobin or methemoglobin can be resolved into two broad lines assignable to intra- and extracellular water which undergoes rapid T2 relaxation by diffusion in magnetic field gradients induced by the intracellular paramagnets. The width of the resolved lines allowed an estimate of the maximum contribution that diffusion makes to T2 relaxation at 7 T. The dependence of the diffusion contribution on the square of the strength of the static magnetic field suggest that diffusion makes a small contribution to water proton T2 relaxation at 1.5 T compared to 7 T, and a negligible one at 0.5 T in early and intermediate hematomas containing deoxyhemoglobin or methemoglobin in intact red blood cells. At the lower field strengths, water proton T2 relaxation is apparently dominated by the rapid chemical exchange (mean lifetime tau = 10 msec) between the intra- and extracellular environments.

We measured the pH of airway surface liquid (ASL) secreted by the ferret trachea in vitro by using a catheter-tipped pH electrode implanted in a collecting cannula close to the airway epithelium. Mucus secretion was promoted by methacholine (0.02 mmol/l) in the organ bath. The pH of the ASL was 6.85 +/- 0.03 (SE) compared with a bath value of 7.39 +/- 0.01, when the bath was bubbled with 5.65% CO2. Changing the bath CO2 from 0 to 20.93% CO2 altered the bath pH from 8.06 to 6.96, but the ASL pH only varied from 6.92 to 6.85. This homeostasis of ASL pH was not the result of the buffering powers of the ASL, because ex situ buffer curves for secreted ASL were similar to those for Krebs-Henseleit solution. Changing the luminal CO2 content by blowing gases through the trachea changed ASL pH by values similar to that ex situ. However, when external organ bath CO2 was changed, the luminal CO2 changes were proportionately far smaller. Measurement of rates of diffusion of CO2 across the tracheal wall indicated that this was not a limiting factor in the results. Similarly, measurement of metabolic rate CO2 production in the tracheal lumen indicated that this did not significantly affect the results. We conclude that the pH of ASL is significantly on the acid side of the pH or interstitial fluid and plasma and that it is maintained relatively constant despite large changes in external pH.

The time course of appearance of morphine and codeine in beard after single dose administration in two human subjects was monitored by radioimmunoassay and confirmed by gas chromatography/mass spectrometry. Both morphine and codeine appeared in beard approximately 7-8 days after drug administration at a time when drug levels in urine, plasma, and saliva were not detectable and drug-induced effects had disappeared. Drug levels in beard appeared to be dose-related suggesting that hair analysis can provide evidence of time and degree of drug exposure.

An ultramicrofluorometric technique was used to analyse the nutrient composition of mouse oviduct fluid. The concentrations of pyruvate, glucose and lactate in the vicinity of the cumulus mass were 0.37, 3.40 and 4.79 mM respectively. In the absence of cumulus cells, the concentration of pyruvate was significantly reduced, to 0.14 mM, while the concentration of glucose was significantly increased to 5.19 mM. Glutamine, which may help to overcome the '2-cell block' in mouse embryos in culture, was present at a concentration of 0.20 mM. A modified medium (MTF) in which the concentration of nutrients was similar to that in mouse oviduct fluid was prepared and its effects on embryo development and metabolism in vitro were compared with that of a conventional embryo culture medium (M16). The percentage of zygotes forming blastocysts in vitro by Day 5 was similar in both media (82% in M16, 79% in MTF). Rates of development, as assessed by cell number, were also comparable. However, the proportion of glucose consumed which was converted to lactate increased dramatically following culture; from 44% in fresh blastocysts, to 73% and 91% in blastocysts derived from 8-cell embryos cultured for 24 h in media MTF and M16 respectively.

"High-performance" liquid-chromatographic (HPLC) methods have been developed for identifying 1-substituted 2-alkyl-3-hydroxypyrid-4-one iron chelators in serum and urine. Ion pairing with heptane- or octanesulfonic acid in pH 2.0-2.2 phosphate buffer and reversed-phase chromatography were required to separate these compounds from endogenous compounds in both biological fluids. In both the 2-methyl and 2-ethyl series of 1-substituted compounds (H, methyl, ethyl, or propyl) the elution times increased in accordance with the n-octanol/water partition coefficients (propyl greater than ethyl greater than H greater than methyl). Urine samples were filtered (0.4 microns pore size) and injected either undiluted or after dilution with elution buffer. After the addition of internal standard, the plasma or serum samples were deproteinized by treatment with HCIO4, 0.5 mol/L, centrifuged, and the supernates were injected directly onto the HPLC. Using these procedures, we could identify 1,2-dimethyl-3-hydroxypyrid-4-one (L1) in the serum and urine of a thalassemic patient who had received a 3-g dose of the drug and in the urine of other patients who had received the same dose. One or more possible metabolites were also observed in the chromatograms of both urine and serum. The 24-h urinary output of L1 (0.22-2.37 g) and iron (10.6-71.5 mg) varied but there was no correlation between the two with respect to quantity or concentration. Instead, urinary iron output was higher in patients with a greater number of transfused units of erythrocytes. This is the first study in humans to show that L1 is absorbed from the gut, enters the circulation, and is excreted in the urine.

This study used CA125, carcinoembryonic antigen (CEA), and alpha-fetoprotein (AFP) to classify ovarian cysts by measuring the levels of the three antigens; this information was useful when fluid obtained through laparoscopic puncture of ovarian cysts was submitted for cytologic examination from patients for whom tissue was unavailable for classification. We studied 136 consecutive cyst fluids (108 benign, 28 malignant) and correlated the findings with the tissue diagnosis. All three antigens were very low (CEA, less than 0.5 ng/ml; CA125, 55-2,143 mu/ml; and AFP, less than 4.8 ng/ml) in follicular and lutein cysts. Markedly elevated CA125 (296-1,950,000 mu/ml) and low CEA (0.5-220 ng/ml) and AFP (less than 4.8 ng/ml) levels were seen in patients with serous neoplasms, both benign and malignant. Elevated CEA (greater than 600 ng/ml) and CA125 (56-65,330 mu/ml) levels were seen in primary mucinous cystadenoma and cystadenocarcinoma. Two patients with colonic carcinoma metastatic to the ovary had an elevated CEA (greater than 600 ng/ml) and a normal CA125. Only one patient, with a malignant teratoma, had an elevated AFP. The adjunctive use of CEA and CA125 is recommended for the classification of ovarian cysts.