Two measures of systemic inflammation are positively associated with haemoglobin levels in adolescent girls living in rural India: a cross-sectional study.

OBJECTIVE: This study tested the hypothesis that systemic inflammation is inversely associated with haemoglobin levels in adolescent girls in India. METHODS: The study population consisted of adolescent girls aged between 10 and 19 years living in a remote rural region in Maharashtra State, India. Data were collected on anthropometric measures, and a venous blood sample was taken and tested for complete blood count and C-reactive protein (CRP). RESULTS: Of 679 individuals who were invited to the research site to participate, data were available from 401 participants giving a response rate of 59%. Median blood CRP was 1.26 mg/l (Range 0.00 to 26.33), and 167 (41.6%) participants had CRP level < 1.0 mg/l. The mean haemoglobin was 12.24 g/dl (standard deviation [SD] 1.51), and the mean total white blood cells (WBC) count was 9.02 × 103 /µl (SD 2.00). With each g/dl increase in blood haemoglobin, the risk of having an elevated CRP of ≥ 1 mg/l increased with an odds ratio of 1.16 (95% CI 1.01 to 1.33, P = 0.03). Total WBC count was also positively associated with blood haemoglobin, increasing by 0.24 × 103 /µl (95% CI 0.11 to 0.37, P < 0.001) per g/dl increase in haemoglobin. Both analyses were adjusted for age. CONCLUSIONS: In this population, blood haemoglobin levels were positively associated with two measures of systemic inflammation, contrary to the primary hypothesis being tested. Other unmeasured environmental exposures may modify haemoglobin levels in this population. Understanding this observation may help design better public health interventions to improve the well-being of adolescent girls in India.

OBJECTIF: Cette étude a testé l'hypothèse selon laquelle l'inflammation systémique est inversement associée aux taux d'hémoglobine chez les adolescentes en Inde. MÉTHODES: La population étudiée était composée d'adolescentes âgées de 10 à 19 ans vivant dans une région rurale éloignée de l'Etat du Maharashtra, en Inde. Les données ont été collectées sur des mesures anthropométriques et un échantillon de sang veineux a été prélevé et testé pour la formule globulaire complète et la protéine C-réactive (CRP). RÉSULTATS: Sur 679 personnes qui ont été invitées au site de recherche à participer, des données étaient disponibles pour 401 participantes, soit un taux de réponse de 59%. La CRP sanguine médiane était de 1,26 mg/L (intervalle de 0,00 à 26,33) et 167 participantes (41,6%) avaient un taux de CRP <1,0 mg/L. L'hémoglobine moyenne était de 12,24 g/dL (écart-type [ET] 1,51), et le nombre moyen total de globules blancs (GB) était de 9,02 x103 /µL (ET 2,00). Avec chaque augmentation par g/dL de l'hémoglobine sanguine, le risque d'avoir une CRP élevée ≥1 mg/L augmentait avec un rapport de cotes de 1,16 (IC95%: 1,01 à 1,33, p = 0,03). La numération totale des GB était également positivement associée à l'hémoglobine sanguine, augmentant de 0,24 x103 /µL (IC95%: 0,11 à 0,37, p <0,001) par g/dL d'augmentation de l'hémoglobine. Les deux analyses ont été ajustées en fonction de l'âge. CONCLUSIONS: Dans cette population, les taux d'hémoglobine sanguine étaient positivement associés à deux mesures de l'inflammation systémique, contrairement à l'hypothèse principale testée. D'autres expositions environnementales non mesurées peuvent modifier les taux d'hémoglobine dans cette population. Comprendre cette observation peut aider à concevoir de meilleures interventions de santé publique pour améliorer le bien-être des adolescentes en Inde.

Cosegregation of the renin allele of the spontaneously hypertensive rat with an increase in blood pressure.

The spontaneously hypertensive rat (SHR) exhibits alterations in the renin-angiotensin-aldosterone system which are similar to those that characterize patients with "nonmodulating" hypertension, a common and highly heritable form of essential hypertension. Accordingly, we determined whether the inheritance of a DNA restriction fragment length polymorphism (RFLP) marking the renin gene of the SHR was associated with greater blood pressure than inheritance of a RFLP marking the renin gene of a normotensive control rat. In an F2 population derived from inbred SHR and inbred normotensive Lewis rats, we found the blood pressure in rats that inherited a single SHR renin allele to be significantly greater than that in rats that inherited only the Lewis renin allele. To the extent that the SHR provides a suitable model of "nonmodulating" hypertension, these findings raise the possibility that a structural alteration in the renin gene, or a closely linked gene, may be a pathogenetic determinant of increased blood pressure in one of the most common forms of essential hypertension in humans.

Factors influencing the survival of rat brain tumor cells after in vitro treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea.

The shape of dose-response curves obtained for asynchronous, exponentially growing 9L rat brain tumor cells treated in vitro with 1,3-bis(2-chloroethyl)-1-nitrosourea changed as a function of the drug exposure time. For short treatment times (less than 1 hr), the dose-response curves had shoulders, indicating that the cells may accumulate sublethal damage; however, after longer treatments (greater than 1 hr), little if any shoulder was apparent. The slope of the exponential portion of the dose-response curve increased progressively with treatment periods from 15 min to 2 hr. Longer exposure times (up to 24 hr) produced no further changes in the cell-kill kinetics. Cell survival was directly related to the BCNU exposure dose [o integral t Co(t)dt] and to the amount of bound BCNU per cell. Extrapolation of the curves for these two variables indicated that some BCNU damage accumulates before death occurs. The amount of serum and cell products available in the medium to bind BCNU affected the level of survival; however, there was no evidence that extracellular spontaneous breakdown products or chemical transformation products were involved in the cell-killing mechanism.

Pharmacokinetics of topically applied sparfloxacin in rabbits.

PURPOSE: Fluoroquinolones are antimicrobial agents that have a broad spectrum of activity and are widely used against many of the ocular pathogens, responsible for conjunctivitis, blepharitis, corneal ulcers etc. The aim of our study was to evaluate the ocular pharmacokinetics of sparfloxacin (0.3% w/v) in the aqueous humour of rabbits. MATERIALS AND METHODS: Pharmacokinetics of topically administered sparfloxacin were determined after a single application of 50 microl topically. The aqueous humour samples were collected at 0, 0.25, 0.5, 1, 2, 3, 4, 5 or 6 hours after instillation. High Performance Thin Layer Chromatographic method was used to analyse the drug concentration in the aqueous humour samples. RESULTS: Fifteen minutes after the instillation of 50 microl of sparfloxacin 0.3% solution, the mean concentration in aqueous humour was found to be 1.4 microg/ml, which reaches the peak level of 3.7 microg/ml after 1.3 hours. At 6 hours, the sparfloxacin aqueous levels were 0.562 microg/ml. The clinical efficacy was predicted based on the Maximum Concentration (Cmax): Minimum Inhibitory Concentration (MIC) and Area Under the Concentration-time curve (AUC):MIC ratios. CONCLUSION: The sparfloxacin levels in aqueous humour of rabbits are sufficiently high up to the 6 hours after instillation in the conjunctival sac to provide bactericidal effect against most of the ocular pathogens. Both Cmax:MIC and AUC:MIC ratios are high enough to provide bactericidal effect against most of the ocular pathogens. Sparfloxacin (0.3%) ophthalmic preparation has excellent penetration through cornea.

Molecular evidence of genetic heterogeneity in Wistar-Kyoto rats: implications for research with the spontaneously hypertensive rat.

Spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto ("WKY") rats are frequently employed in experimental studies of hypertension. Although both SHR and WKY rat strains have been presumed to be fully inbred, recent studies have revealed important biologic variability in WKY rats from different commercial sources. Genealogic evidence suggests that, in the United States, breeding stocks of WKY rats may have been distributed to major commercial suppliers as early as the F10 generation. To test the hypothesis that commercially available WKY rats are genetically heterogeneous, we performed deoxyribonucleic acid (DNA) "fingerprint" analysis on genomic DNA of WKY rats from two of the largest vendors in the United States, Taconic Farms and Charles River Laboratories. We found molecular evidence of genetic variability not only among WKY rats from two different breeding facilities, but also among WKY rats within a single breeding facility (Taconic Farms). Although some studies have suggested the possibility of biologic variability in SHR from different sources, preliminary studies have not revealed molecular evidence of genetic heterogeneity in SHR from these vendors. In demonstrating genetic variability in WKY rats from different sources, the current study provides compelling evidence that rats designated WKY do not constitute an inbred strain. Accordingly, the results of studies in which SHR and WKY rats are compared might vary because of genetic heterogeneity in "the WKY rat control strain."

Urinary catecholamines in iron-deficient rats at rest and following surgical stress.

The purpose of this study was to determine catecholamine concentrations both at rest and in response to a surgical stress in iron-deficient and control rats. Twenty-one-day-old rats were randomized to one of two groups which received a diet containing either 6 or 50 mg iron/kg. Three to five days later, when anemia was first detectable, urinary norepinephrine (NE) concentrations were already significantly elevated in the iron-deficient compared to control rats. In contrast, urinary dopamine (DA) became depressed after 10 days of the iron-deficient regimen. At 38 days of age, both groups were subjected to a surgical stress. NE and DA became elevated over baseline values in both diet groups during the 24-h period following surgery; NE remained significantly higher and DA significantly lower in the iron-deficient than in the control group. We conclude that changes in urine catecholamine concentration occur early in the development of iron deficiency and that they are characteristic of both baseline and stress conditions.

Relationship of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) pharmacokinetics of uptake, distribution, and tissue/plasma partitioning in rat organs and intracerebral tumors.

To obtain a clearer definition of the relationship between the structure of BCNU and CCNU and their antitumor activity, we determined the uptake, distribution, and tissue/plasma partition ratios of both compounds in normal organs and intracerebral (ic) 9L tumors in rats. Greater uptake, distribution, and tissue/plasma partition ratios were obtained for parent CCNU in fat, liver, and brain, and for parent BCNU in kidney. CCNU distributes more rapidly and extensively than BCNU only in fatty tissues. BCNU distributed more extensively in kidney and liver. Rats received bolus IV injections of 14C-labeled BCNU or CCNU in increasing doses; measurements taken 30 min after injection showed that three- to fourfold more BCNU than CCNU was bound to nucleic acids in brain and ic 9L tumor tissue. Because the chloroethyl group is the alkylating moiety for both drugs, these findings implied that BCNU biotransformed to its reactive intermediate more rapidly than did CCNU. These observations, together with previous findings, indicate that one reason for the greater effectiveness of BCNU than CCNU against ic 9L tumors is its superior ability to form an intermediate that can bind to ic 9L tumor cell nucleic acids.

Liquid chromatography with fluorometric detection of unconjugated estriol in serum of pregnant women.

We describe a fluorometric liquid chromatographic assay for the measurement of unconjugated estriol in the serum of pregnant women. Estriol is extracted into methylene chloride/propanol-2 from serum by use of a Clin-Elut extraction column, the extract evaporated, and the residue redissolved in mobile phase. An aliquot is injected onto the liquid chromatograph and the estriol is separated on a reversed-phase octyl column with a mobile phase consisting of acetonitrile/phosphate buffer (23:77, v/v). The effluent is monitored by fluorescence detection. The proposed method offers good reproducibility (CV less than 7%), sensitivity (less than 0.5 micrograms/l), and accuracy. Of many drugs and steroids tested, only 16,17-epiestriol interferes with the estriol analysis.

Simultaneous liquid chromatographic analysis of amitriptyline, nortriptyline, imipramine, desipramine, doxepin, and nordoxepin.

A simultaneous method for the therapeutic monitoring of amitriptyline, doxepin, imipramine, and their active demethylated metabolites nortriptyline, nordoxepin, and desipramine, respectively, in plasma or serum by reversed-phase liquid chromatography (RPLC) is presented. The drugs and the internal standard (loxapine) are first extracted from 2 ml of serum into butylchloride at pH 14, and then back extracted into 200 microliter of 0.025 mol/l hydrochloric acid. An aliquot of the aqueous acid phase is injected into the chromatograph and eluted with acetonitrile-phosphate buffer (21: 79, by vol.) containing 0.6 nl of n-nonylamine per liter of phosphate buffer. The drugs are eluted in a total chromatographic time of approximately 13 min at ambient temperature and detected at 200 nm. A sensitivity of 5 microgram/l of serum for each drug is obtained. Recoveries for these drugs ranged from 77% to 103%; and the coefficient of variation (day-to-day) ranged from 4.2 to 7.8. Of 35 basic or neutral drugs tested for possible interference, only propoxyphene interferes with the analysis of nortriptyline.

Pharmacokinetics of intracarotid artery 14C-BCNU in the squirrel monkey.

A comparison of intravenous to intracarotid artery (ICA) administration of 14C-BCNU (1,3-bis(2-chloroethyl)-1-nitrosourea) was made in squirrel monkeys. Radioactivity was measured as soluble drug products and as RNA-, DNA-, and protein-bound radioactivity. The ICA administration of BCNU achieved 190% to 280% higher brain nucleic acid-bound drug levels than use of the intravenous route in the infused hemisphere and 130% to 280% higher levels than in the noninfused hemisphere. In addition, some brain regions directly subserved by the middle cerebral artery had bound drug levels four- to fivefold greater than those found in regions of noninfused brain. The data suggest that a need for BCNU dose reduction due to myelotoxicity may be an indication for ICA therapy in selected brain-tumour cases.

Simple analogs of the toxin callicarpone.

Callicarpone, a component 10 times as toxic to fish as rotenone, has been isolated from the leaves of Callicarpa candicans. It is reasonable to assume that callicarpone will act as an insecticidal agent as does rotenone. Therefore, the structure-activity relationship of callicarpone was examined by synthesizing a series of compounds having certain of its structural features. Those compounds were tested for insecticidal and antimicrobial activities. A study of synthetic analogs elucidated the functional group chemistry of callicarpone so that a synthesis might be undertaken. Piperitone oxide showed approximately 1/100th the activity of rotenone against Daphnia magna. 1-(alpha-Hydroxyisopropyl)-3-oxocyclohexene oxide showed activity against myobacterium while 2,3,4,6,7,8-hexahydronaphthalene-1,4-dione showed inhibitory activity against the mycobacterium and two yeasts.

Liquid chromatographic determination of gentamicin in serum with spectrophotometric detection.

A simple, accurate, and specific liquid chromatographic procedure is described for the determination of gentamicin in 50 muL of serum. Gentamicin and sisomicin (IS) were converted into their trinitrophenyl derivatives by reaction with a water soluble derivatizing agent at 70 degrees C for 30 minutes. The derivatives were extracted from the crude mixture with chloroform, which was separated, evaporated, and the residue reconstituted in acetonitrile. Gentamicin was separated into three major isomers (C1, C2, and C1a) on a reversed-phase octyl column by eluting with a mobile phase. The eluted compounds were detected at 340 nm and quantified from their peak heights or peak areas. Chromatography was complete in less than 11.0 minutes. The lower limit of detection for gentamicin was less than 0.5 mg/L, analytical recoveries varied from 96.6% to 99.3%, linearity extended to 15.0 mg/L, and day-to-day precision was between 2.2% to 2.9%.

Rapid method for screening toxic drugs in serum with liquid chromatography.

We present a method for the simultaneous analysis of a variety of commonly abused drugs (acetaminophen, theophylline, salicylate, primidone, methyprylon, phenobarbital, butabarbital, ethchlorvynol, butalbital, chlordiazepoxide, pentobarbital, amobarbital, phenytoin, glutethimide, secobarbital, flurazepam, nitrazepam, methaqualone, N-desmethyldiazepam, and diazepam) in serum or plasma. Serum proteins are precipitated with an acetonitrile solution containing hexobarbital, the internal standard. The drugs are eluted from a reversed-phase column with a mobile phase consisting of acetonitrile/phosphate buffer (pH 3.2), using a two-step linear gradient, at a flow rate of 3.0 mL/min. The eluted drugs are detected by their absorption at 210 nm; their quantities are estimated from their peak heights. A complete analysis requires no longer than 45 minutes at the optimum column temperature of 50 degree C. A sensitivity of 2 mg/L of serum is attained routinely for most of the hypnotic and analgesic drugs; while methaqualone, chlordiazepoxide, diazepam, and N-desmethyldiazepam can be detected at a concentration of 0.2 mg/L. Analytical recoveries for the twenty drugs varied from 93-112%, with good reproducibility. Of more than forty drugs tested for possible interference, desmethyldoxepin, procainamide, phenylpropanolamine, mesantoin, and phenacetin interfere with the analysis of flurazepam, acetaminophen, ethchlorvynol, and phenobarbital, respectively.

Extraintestinal manifestations of idiopathic ulcerative colitis in northwestern India.

OBJECTIVE: To determine the frequency of extraintestinal manifestations in patients with idiopathic ulcerative colitis. METHODS: 46 patients underwent detailed clinical, biochemical and radiological evaluation. RESULTS: One patient (2%) had peripheral arthritis and two patients (4%) had ocular involvement in the form of anterior uveitis. No patient had mucocutaneous, vascular, or hepatobiliary manifestations, or sacroiliitis. CONCLUSIONS: The frequency of extraintestinal manifestations in patients with IUC in northwestern India is low.

Clinical analysis of individual steroids by column liquid chromatography.

At present, there are various LC methods available for the determination of estriol, aldosterone, and cortisol in urine, and for cortisol, cortisone, 11-deoxycortisol, corticosterone, 17-hydroxyprogesterone, estriol, vitamin D isomers, and various exogenous glucocorticoids in serum. The LC methods are more specific than the currently available RIAs or homogenous immunoassays. However, whether the data obtained by more specific LC methods are better clinically than the commonly used immunoassays for these steroids must still be proven. In this review, I have critically evaluated various LC methods currently available for the routine determination of clinically important steroids in the clinical laboratories. A complete evaluation and advantages and disadvantages of alternative techniques are beyond the scope of this review. However, the readers are urged to refer to the review articles and chapters listed in the reference section.

Liquid chromatographic determination of cyclosporine in whole blood with the advanced automated sample processing unit.

We describe a rapid, precise, cost-effective, and accurate isocratic liquid chromatographic (LC) procedure for determining cyclosporine in whole blood. The cyclosporine is extracted from 0.5 ml of whole blood together with 200 micrograms of cyclosporin D, added per liter as internal standard, by using an Advanced Automated Sample Processing (AASP) unit. The on-line solid-phase extraction is performed on an octasilane sorbent cartridge which is interfaced with a Perkin-Elmer 83 X 4.6 mm I.D. cartridge column, packed with 3-micron octadecyl packing. The column is eluted with a mobile phase containing acetonitrile-water (13:7) at a flow-rate of 1.0 ml/min at a column temperature of 70 degrees C. The column effluent is monitored at 210 nm. The absolute recovery of cyclosporine exceeded 87% and the linearity extended up to 2000 micrograms/l. Within-run and day-to-day coefficients of variation were less than 8%. The correlation between AASP-LC and manual Bond-Elut extraction-LC method was excellent (r = 0.97).