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Calcimimetics activate the calcium-sensing receptor (CAR) and reduce parathyroid hormone (PTH) by increasing the sensitivity of the parathyroid CaR to ambient calcium. The calcimimetic, cinacalcet, is effective in treating secondary hyperparathyroidism in dialysis patients [chronic kidney disease (CKD 5)], but little is known about its effects on stage 3-4 CKD patients. We compared cinacalcet and paricalcitol in uremic rats with creatinine clearances 'equivalent' to patients with CKD 3-4. Uremia was induced in anesthetized rats using the 5/6th nephrectomy model. Groups were 1) uremic control, 2) uremic + cinacalcet (U+Cin; 15 mg x [kg.sup.-1] x [day.sup.-1] po for 6 wk), 3) uremic + paricalcitol (U+Par; 0.16 [micro]g/kg, 3 x wk, ip for 6 wk), and 4) normal. Unlike U+Par animals, cinacalcet promoted hypocalcemia and marked hyperphosphatemia. The Ca x P in U+Cin rats was twice that of U+Par rats. Both compounds suppressed PTH. Serum 1,25-[(OH).sub.2][D.sub.3] was decreased in both U+Par and U+Cin rats. Serum FGF-23 was increased in U+Par but not in U+Cin, where it tended to decrease. Analysis of tibiae showed that U+Cin, but not U+Par, rats had reduced bone volume. U+Cin rats had similar bone formation and reduced osteoid surface, but higher bone resorption. Hypocalcemia, hyperphosphatemia, low 1,25-[(OH).sub.2][D.sub.3], and cinacalcet itself may play a role in the detrimental effects on bone seen in U+Cin rats. This requires further investigation. In conclusion, due to its effects on bone and to the hypocalcemia and severe hyperphosphatemia it induces, we believe that cinacalcet should not be used in patients with CKD without further detailed studies. calcimimetics; vitamin D; hyperphosphatemia; uremia; bone formation and bone resorption doi: 10.1152/ajprenal.00552.2009.

Calorimetric and spectroscopic techniques are used to analyze the Ti(IV) interactions with transferrin and the model N,N'-di(o-hyroxybenzyl)ethylenediamine-N,N'-diacetic acid (HBED). Results reveal that Ti(IV)-bound transferrin nominally enhances the thermal stability.

Apolipoprotein L-I (apoL-I) is a human high-density lipoprotein (HDL) component able to kill Trypanosorna brucei brucei by forming anion-selective pores in the lysosomal membrane of the parasite. Another HDL component, haptoglobin-related protein (Hpr), has been suggested as an additional toxin required for full trypanolytic activity of normal human serum. We recently reported the case of a human lacking apoL-I ([apoL-I.sup.-/-]HS) as the result of frameshift mutations in both apoL-I alleles. Here, we show that this serum, devoid of any trypanolytic activity, exhibits normal concentrations of HDL-bound Hpr. Conversely, the serum of individuals with normal HDL-bound apoL-I but who lack Hpr and haptoglobin [[Hp(r).sup.-/-]HS] as the result of gene deletion (anhaptoglobinemia) exhibited phenotypically normal but delayed trypanolytic activity. The trypanolytic properties of [Hp(r).sup.-/-]HS were mimicked by free recombinant apoL-I, whereas recombinant Hpr did not affect trypanosomes. The lysis delay observed with either [Hp(r).sup.-/-]HS or recombinant apoL-I could entirely be attributed to a defect in the uptake of the lytic components. Thus, apoL-I is responsible for the trypanolytic activity of normal human serum, whereas Hpr allows fast uptake of the carrier HDL particles, presumably through their binding to an Hp/Hpr surface receptor of the parasite. innate immunity | sleeping sickness | rypanolytic factor | haptoglobin receptor | Trypanosoma brucei

Capture and detection of immunoglobulin E (IgE) in simple solution and in human serum using an aptamer-modified probe surface for affinity matrix-assisted laser desorption/ionization mass spectroscopy detection is reported. Detectable signals were obtained for 1 amol of IgE applied either in a single, 1-[micro]L application of 1 pM IgE or after 10 successive, 1-/[micro]L applications of 100 fM IgE. In both cases, the surface was rinsed after each application of IgE to remove sample concomitants including salts and free or nonspecifically associated proteins. Detection of native IgE, which is the least abundant of the serum immunoglobulins and occurs at subnanomolar levels, in human serum was demonstrated and interference from the high-abundance immunoglobulins and albumin was investigated. The aptamer-modified surface showed high selectivity toward immunoglobulins in serum, with no significant interference from serum albumin. Addition of IgE to the serum suppressed the signals from the other immunoglobulins, confirming the expected selectivity of the aptamer surface toward IgE. Dilution of the serum increased the selectivity toward IgE; the protein was detected without interference in a 10 000-fold dilution of the serum, which is consistent with detection of IgE at amol (pM) levels in standard solutions.

We report an electrochemical, aptamer-based (E-AB) sensor for the detection of platelet-derived growth factor (PDGF) directly in blood serum. The E-AB approach employs alternating current voltammetry to monitor target-induced folding in a methylene blue-modified, PDGF-binding aptamer. The sensor is sensitive, highly selective, and essentially reagentless: we readily detect the BB variant of PDGF at 1 nM directly in undiluted, unmodified blood serum and at 50 pM (1.25 ng/mL) in serum-diluted 2-fold with aqueous buffer. The sensitivity and selectivity achieved by this sensor match or significantly exceed those of the best analogous optical approaches. For example, the detection limit attained in 50% serum is achieved against a > 25 million-fold excess of contaminating blood proteins and represents a 4 order of magnitude improvement over the most sensitive optical PDGF aptasensor reported to date. Moreover, the E-AB sensor combines these promising attributes in a platform that is reusable, label-free, and electronic. Given these advantages, E-AB sensors appear well suited for implementation in portable microdevices directed at the direct detection of proteins and small molecules in complex, largely unprocessed clinical samples.

This narrative review examines serum biomarkers for the diagnosis and monitoring of delirium. Serum biomarkers for delirium fall into three major groups: 1) those that are present or elevated prior to disease onset--risk markers, 2) those that rise with onset and fall with recovery--disease markers, and 3) those that rise in proportion to the consequences of disease--end products. As risk markers, we examine serum chemistries and genetic risk markers. As disease markers, we examine serum anticholinergic activity, amino acids, melatonin, cytokines, cortisol, and gene expression. As end products of delirium, we examine markers of neuronal injury. Finally, we discuss methodological and biostatistical considerations for future biomarker studies. Identifying accurate biomarkers for delirium may shed further light into its pathophysiology and on the interrelationship between delirium and dementia.

Progesterone is a steroid hormone that is involved in regulating female reproductive processes. Its concentration in blood is measured to determine ovarian function. A candidate reference measurement procedure for progesterone in human serum involving isotope dilution coupled with liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been developed and critically evaluated. The progesterone along with its internal standard (progesterone-[sup.13][C.sub.2]) was extracted from the serum matrix using liquid--liquid extraction prior to reversed-phase LC/MS/ MS. The accuracy of the measurement was evaluated by a comparison of results of this method on a lyophilized human serum reference material for progesterone [Certified Reference Material (CRM) 347] with the certified values determined by gas chromatography/mass spectrometry (GC/MS) reference methods and by a recovery study for the added progesterone. The results of this method for progesterone agreed with the certified value within the uncertainty of the measurements for the CRM 347. The recovery of the added progesterone ranged from 100.1 to 100.9%. This method was applied to the determination of progesterone in frozen serum samples from three individual female donors with the progesterone concentrations ranging from 0.151 to 24.42 ng/g. Excellent reproducibility was obtained with within-set coefficients of variation (CVs) ranging from 0.1 to 1.4%, and between-set CVs ranging from 0.3 to 0.5%. Excellent linearity was also obtained with correlation coefficients of all linear regression lines (measured intensity ratios vs mass ratios) ranging from 0.9998 to 1.0000. The detection limit at a signal-to-noise ratio of ~3 was 1.8 pg of progesterone. This well-characterized LC/MS/MS method for serum progesterone, which demonstrates good accuracy and precision, low susceptibility to interferences, and comparability with GC/MS reference methods, qualifies as a reference measurement procedure that can be used to provide an accuracy base to which routine methods for progesterone can be compared and that will serve as a standard of higher order for measurement traceability.

Protein glycosylation has been implicated in key biological processes including immunological recognition, cellular adhesion, protein folding, and signaling as well as disease progression. Although several methods are available to assess glycosylation of protein structures, none of them is able to screen complex biological samples at a global as well as an individual scale. A novel strategy presented here uses an all-liquid phase enrichment and prefractionation methodology coupled to glycoprotein microarray technology using a multiple lectin-based, biotin--streptavidin detection scheme. Selective detection of glycan structures was made possible by employing multiple lectins to screen glycoprotein standards as well as serum samples from normal subjects or patients with chronic pancreatitis or pancreatic cancer. Interestingly, in some instances, a greater degree of glycosylation was seen in proteins that were underexpressed based on the reversed-phase chromatogram alone. Studies with standard proteins established the limits of detection to be in the 2.5-5-fmol range. Studies on serum samples showed differences in glycosylation patterns, particularly with respect to sialylation, mannosylation, and fucosylation, in normal, pancreatitis, and cancer sera. By coupling glycoprotein enrichment and fractionation with a microarray platform, we have shown that naturally occurring glycoproteins from human serum can be screened and characterized for different glycan structures, thereby allowing one to do comparative studies that monitor individual glycosylation changes within a glycoproteome representing different biological states. This approach may be useful to identify potential biomarkers in cancer.

Although sample stacking has enjoyed some degree of success in electrophoretic separation techniques, there is still a major problem with complex matrix sample as it suffers tremendously from sample matrix effects. A novel method that combines two concentration techniques, centrifuge microextraction (CME) and on-line back-extraction field-amplified sample injection (OLBE-FASI), is used to determine trace ephedrine derivatives in urine and serum by capillary zone electrophoresis. The CME, integrating the sample cleanup and preconcentration into a single step, is a promising sample preparation method for biological samples. The CME technique provided 9-14-fold enrichment within 10 min. The OLBE-FASI eliminated the need to perform solvent exchange and provided a further concentration of the analytes. Using CME coupled with OLBE-FASI, over a 3800-fold increase in sensitivity could be obtained as compared with the normal hydrodynamic injection without sample stacking. For a 1-mL urine sample, the linear range was 5/10-200 ng/mL with the square of the correlation coefficients ([r.sup.2]) ranging from 0.9988 to 0.9994. Detection limits were from 0.15 to 0.25 ng/mL using a photodiode array UV detection at wavelength 192 nm. The possibility of this method to determine ephedrine derivatives in 20-/[micro]L serum samples was also demonstrated.

Concurrent with the explosion in the number of publications reporting biomarker discovery by profiling technologies, such as proteomics and pattern recognition, has been the increase in evidence highlighting the susceptibility of these approaches to analytical and experimental bias. The work presented here addresses these timely issues by delivering a detailed characterization of the effect of common sources of bias in clinical studies on serum and plasma profiles generated by a key technology in metabonomics, NMR spectroscopy. Specifically, differences in composition when blood samples were collected onto and in the absence of ice, over a series of serum--clot contact times, the stability of NMR-prepared samples over time and the effect on the metabolic profile of freeze--thawing were examined. While differences between individuals were far greater than variation from any other experimental factor, each of the conditions examined did cause slight alterations to the NMR profile that could produce a systematic bias. Variation due to clotting time caused changes in energy metabolites, which were delayed by ice with no other spectral effects. Room-temperature stability and hence NMR spectral repeatability were high (< 1% intrasample variation). Higher molecular weight species such as lipoproteins were more susceptible to the variations present in the examined factors. These observations have implications for profiling study design, and hence, our results form a new and valuable resource for those attempting clinical metabolic profiling, for regulatory agencies involved in the licensing of clinical tests and in the generation of international reporting standards for metabonomics.

A nonlinear alignment strategy was examined for the quantitative analysis of serum metabolites. Two small-molecule mixtures with a difference in relative concentration of 20-100% for 10 of the compounds were added to human serum. The metabolomics protocol using UPLC and XCMS for LC-MS data alignment could readily identify 8 of 10 spiked differences among more than 2700 features detected. Normalization of data against a single factor obtained through averaging the XCMS integrated response areas of spiked standards increased the number of identified differences. The original data structure was well preserved using XCMS, but reintegration of identified differences in the original data reduced the number of false positives. Using UPLC for separation resulted in 20% more detected components compared to HPLC. The length of the chromatographic separation also proved to be a crucial parameter for a number of detected features. Moreover, UPLC displayed better retention time reproducibility and signal-to-noise ratios for spiked compounds over HPLC, making this technology more suitable for nontargeted metabolomics applications.

Benzene is an important industrial chemical and environmental contaminant that causes leukemia. To obtain mechanistic insight into benzene's mechanism of action, we examined the impact of benzene on the human serum proteome in a study of exposed healthy shoe-factory workers and unexposed controls. Two sequential studies were performed, each using sera from 10 workers exposed to benzene (overall mean benzene air level >30 ppm) and 10 controls. Serum samples were subjected to anion-exchange fractionation and bound to three types of ProteinChip arrays (Ciphergen Biosystems, Fremont, CA) [hydrophobic (H50), metal affinity (IMAC3-Cu), and cation exchange (WCX2)]. Protein-expression patterns were detected by surface-enhanced laser desorption/ ionization (SELDI)-TOF MS. Three proteins (4.1, 7.7, and 9.3 kDa) were consistently down-regulated in exposed compared with control subjects in both studies. All proteins were highly inversely correlated with individual estimates of benzene exposure (r > 0.75). The 7.7- and 9.3-kDa proteins were subsequently identified as platelet factor (PF)4 and connective tissue activating peptide (CTAP)-III. Initial proteomic results for PF4 and CTAP-III were subsequently confirmed in a single experiment using a Protein-Chip-array-based immunoassay(Ciphergen Biosystems). The altered expression of the platelet-derived CXC-chemokines (40% and 63% for PF4 and CTAP-III, respectively) could not be explained by changes in absolute platelet counts. Thus, SELDI-TOF analysis of a limited number of exposed and unexposed subjects revealed that lowered expression of PF4 and CTAP-III proteins is a potential biomarker of benzene's early biologic effects and may play a role in the immunosuppressive effects of benzene. biomarker | leukemia | mass spectrometry | platelet | molecular epidemiology

A novel surface-enhanced Raman scattering (SERS) based approach for the quantitative determination of creatinine in human serum is described. Using isotopically labeled (2-[sup.13]C, 2,3-[sup.15][N.sub.2]) creatinine as internal standard, SERS acquires the character of a ratio method that works similar to the well-established isotope dilution techniques. In conjunction with multivariate data analysis, the method was successfully applied for quantifying creatinine at clinically relevant levels and below. A partial least-squares regression model was generated from a set of 87 calibration spectra covering the full range of mole fractions of neat creatinine. The prediction performance of the model was thereafter validated with independent reference samples giving a standard deviation of less than 2%. Finally, a conditioning procedure to prepare real serum samples for SERS-based creatinine analysis was worked out and validated. Measured serum creatinine concentrations are within 3% of the values obtained from gas chromatography/isotope dilution mass spectrometry on the same serum starting material.

Background. Although a host of factors are known to influence 25-hydroxyvitamin D [25(OH)D] serum levels, few studies addressed the distinctive sex-specific influence of aging, and the age-specific relationship of parathyroid hormone (PTH) with 25(OH)D. The aims of this research were to evaluate changes of 25(OH)D and PTH levels with age in a large population-based sample of men and women and to test the hypothesis that 25(OH)D serum concentrations needed to offset age-associated hyperparathyroidism are significantly higher in older than in younger persons. Methods. In 1107 participants of the InCHIANTI (Invecchiare in Chianti, i.e., Aging in the Chianti area) study, we collected information on dietary intake, daylight exposure, and disability, and measured renal function and serum 25(OH)D and PTH. Results. In women, the age-related decline of 25(OH)D was already evident shortly after age 50, whereas in men it started only after age 70 and was substantially less steep. Age, daylight exposure, winter season, and disability were independent predictors of low 25(OH)D levels. For any given level of 25(OH)D, PTH levels were progressively and consistently higher in older than in younger participants. Conclusions. These findings suggest that the age-associated fall of serum 25(OH)D starts earlier in women than in men and that higher levels of 25(OH)D are required in older compared to younger persons to avoid the age-associated compensatory hyperparathyroidism.

A novel N-linked oligosaccharide (N-glycan) with '[beta]1-4 bisecting branch (galactose [beta]1-4 bisecting N-acetylglucosamine)' was found in human serum IgG. Its structure was efficiently analyzed by using [beta]-galactosidase digestion, a [MS.sup.n] spectral library database, and negative-ion [MS.sup.2] spectral matching. For confirmation, the novel N-glycan was synthesized by using an expected standard N-glycan (acceptor), UDP-galactose (donor), and [beta]1-4 galactosyl-transferase. This work also demonstrates that the [MS.sup.n] spectral library database, in particular, negative-ion [MS.sup.2] spectral matching, can efficiently reduce the number of specific, sequential exoglycosidase digestions required and is useful for rapid structural analysis of unknown glycans not in the database.

Kinins are vasoactive peptides released from precursors called kininogens, and serum levels of both T- and K-kinmogens increase dramatically as rats age. Kinin release is tightly regulated, and here we show that serum kinin levels also increase with age, from 63 [+ or -] 16 nmol/L in young Fisher 344 rats to 398 [+ or -] 102 nmol/L in old animals. Both K- and T-kininogens contribute sequentially to this increase, with the increase in middle-aged animals being driven primarily by K-kininogen, whereas the further augmentation in older rats occurs by increasing T-kininogen. By measuring ERK activation, we show that aorta endothelial cells from old animals are hyporesponsive to exogenous bradykinin. However, if serum kinin levels are experimentally decreased by lipopolysaccharide treatment, then the endothelial response to bradykinin is reestablished. These results indicate that serum levels of kinins increase with age, whereas the responsiveness of target cells to kinins is reduced in these same animals.

The objective of this study was to analyze the validity of the velocity corresponding to the onset of blood lactate accumulation (OBLA) and critical velocity (CV) to determine the maximal lactate steady state (MLSS) in soccer players. Twelve male soccer players (21.5 [+ or -] 1.0 years) performed an incremental treadmill test for the determination of OBLA. The velocity corresponding to OBLA (3.5 mM of blood lactate) was determined through linear interpolation. The subjects returned to the laboratory on 7 occasions for the determination of MLSS and CV. The MLSS was determined from 5 treadmill runs of up to 30-minute duration and defined as the highest velocity at which blood lactate did not increase by more than 1 mM between minutes 10 and 30 of the constant velocity runs. The CV was determined by 2 maximal running efforts of 1,500 and 3,000 m performed on a 400-m running track. The CV was calculated as the slope of the linear regression of distance run versus time. Analysis of variance revealed no significant differences between OBLA (13.6 [+ or -] 1.4 km x [h.sup.-1]) and MLSS (13.1 [+ or] 1.2 km x [h.sup.-1]) and between OBLA and CV (14.4 [+ or -] 1.1 km x [h.sup.-1]). The CV was significantly higher than the MLSS. There was a significant correlation between MLSS and OBLA (r = 0.80), MLSS and CV (r = 0.90), and OBLA and CV (r = 0.80). We can conclude that the OBLA can be utilized in soccer players to estimate the MLSS. In this group of athletes, however, CV does not represent a sustainable steady-state exercise intensity. KEY WORDS. aerobic capacity, steady state, athletes, exercise intensity

UDP-glucuronosyltransferase (UGT) 1A1 is a conjugating biotransformation enzyme that plays a role in maintaining levels of endogenous compounds (e.g., bilirubin) and handling exogenous compounds, including carcinogens. The UGT1A1*28 polymorphism results in decreased UGT1A1 promoter activity due to 7 thymine-adenine (TA) repeats instead of the commonly found 6 repeats. Studies indicate that foods from the botanical families Cruciferae (e.g., broccoli), Rutaceae (citrus), Liliaceae (e.g., onions), and Leguminosae (legumes) may increase UGT activity. We investigated, in an observational study, whether foods from these botanical groups were associated with increased UGT1A1 activity as indicated by serum bilirubin concentrations and whether the effect varied by UGT1A1*28 genotype, comparing those homozygous for the [[TA].sub.7]-repeat allele (7/7) to homozygous wild-types (6/6) and heterozygotes (6/7) combined. Healthy volunteers completed 3-d food records. Blood samples were drawn for genomic DNA collection and bilirubin measures. For total, direct, and indirect bilirubin measures, there was no significant association with any botanical group independently. There was a significant inverse association between all 3 bilirubin measures and interaction of UGT1A1*28 genotype with Cruciferae intake (P < 0.02 for each measure); individuals with the 7/7 genotype had reduced bilirubin concentrations with increased intake of cruciferous vegetables, whereas individuals with the 6/6 or 6/7 genotype did not. With regard to UGT1A1-conjugated carcinogens (e.g., heterocyclic amines, polycyclic aromatic hydrocarbons), individuals with decreased UGT1A1 activity due to the 7/7 genotype may be at greater risk for carcinogenesis, but our results imply that they also may have greater opportunity to decrease that risk through dietary intervention. KEY WORDS: * UGT1A1 * polymorphism * chemoprevention * Cruciferae * bilirubin

A study is conducted to determine the serum and gingival crevicular fluid (GCF) concentrations of systemic ciprofloxacin in patients with periodontitis. Results reveal that ciprofloxacin is found in GCF and its concentration in GCF is significantly higher than that in serum.

A new method using comprehensive two-dimensional gas chromatography and isotope dilution time-of-flight mass spectrometry (GC x GC-IDTOFMS) for the simultaneous measurement of selected polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), and brominated flame retardants is presented. In contrast to the reference methods based on classical GC/MS, a single injection of the extract containing all compounds of interest results in accurate identification and quantification. Using GC x GC ensures the chromatographic separation of most compounds, and TOFMS allows mass spectral deconvolution of coeluting compounds as well as the use of [sup.13]C-labeled internal standards for quantification. Isotope ratio measurements of the most intense ions for both native and labels ensure the required specificity. The use of this new method with an automated sample preparation procedure developed at the Centers for Disease Control and Prevention (CDC) for the analysis of human serum and milk compared favorably to conventional isotope-dilution one-dimensional gas chromatography-high-resolution mass spectrometry (GC-IDHRMS) for the different human serum and milk pools tested. The instrumental detection limits ranged between 0.5 pg/[micro]L and 10 pg/[micro]L and the method detection limits ranged between 1 and 15 pg/[micro]L (N = 59 analytes). The reproducibility of the method was almost as good as with GC-IDHRMS, the relative standard deviations ranging between 1 and 11% for OCPs measured in human serum. OCP, PBDE, and PCB levels measured using the two methods were highly correlated, and the deviations between the two methods were below 20% for most analytes with concentrations above 1 ng/g milk lipids.

Byline: P. P. Golikov (1), N. Yu. Nikolaeva (1), B. V. Davydov (1), E. V. Klychnikova (1), A. N. Pogodina (1), E. B. Nikolaeva (1), M. M. Abakumov (1) Abstract: The correlations between nitric oxide products (NO.sub.x) and the angiotensin-converting enzyme (ACE) activity and malondialdehyde (MDA) were studied in the blood serum of patients with thoracic wounds (the test group) 1, 3, 7, and 14 days after the injury. The severity of a patient's state scored from 8 to 15 on the APACHE II scale. The control group comprised 20 blood donors. One day after wounding, NO.sub.x, MDA, and ACE activity significantly increased. Subsequently, NO.sub.x and ACE activity decreased, whereas MDA increased. A significant negative correlation was found between NO.sub.x and MDA, and a significant positive correlation, between NO.sub.x and ACE activity. The correlations observed in the control group were undetectable in the patients one day after wounding but were restored on the third day. Thus, thoracic injury was accompanied by an increase in NO.sub.x and MDA, which determine the severity of oxidative stress. The positive correlation between NO.sub.x and ACE activity in the control and test groups reflects their concerted action in regulating the physiological functions and metabolism. Author Affiliation: (1) Sklifosovskii Institute of Emergency Medical Care, Moscow, Russia Article History: Registration Date: 28/12/2004

The determinants of blood levels of carotenoids were previously investigated in small or select samples. The relations of serum lutein + zeaxanthin to possible diet, lifestyle, and physiological determinants in 7059 participants of the Third National Health and Nutrition Examination Survey (1988-1994), [greater than or equal to] 40 y old, were examined. In a fully adjusted, multiple linear regression model, lower serum lutein + zeaxanthin was significantly associated with smoking, heavy drinking, being white, female, or not being physically active, having lower dietary lutein + zeaxanthin, higher fat-free mass, a higher percentage of fat mass, a higher waist-hip ratio, lower serum cholesterol, a higher white blood cell count, and high levels of C-reactive protein (P < 0.05). The model explained 24% of the variation present in serum lutein + zeaxanthin for the current sample. The correlation between dietary and serum lutein + zeaxanthin was 0.17 and increased to 0.18 after adjusting for the effects of given covariates. Each 10% increase in dietary lutein + zeaxanthin was associated with a 1% increase in serum conditional on other terms in the model. Many factors that influence the level of serum lutein + zeaxanthin remain unknown. KEY WORDS: * body mass * carotenoids * diet lutein * smoking * zeaxanthin

This study quantifies the experimental uncertainty for LC retention time, mass measurement precision, and ion abundance obtained from replicate nLC-dual ESI-FF-ICR analyses of the low molecular weight fraction of serum. We used ultrafiltration to enrich the < 10-kDa fraction of components from the high-abundance proteins in a pooled serum sample derived from ovarian cancer patients. The THRASH algorithm for isotope cluster detection was applied to five replicate nLC-dual ESI-FT-ICR chromatograms. A simple two-level grouping algorithm was applied to the more than 7000 isotope clusters found in each replicate and identified 497 molecular species that appeared in at least four of the replicates. In addition, a representative set of 231 isotope clusters, corresponding to 188 unique molecular species, were manually interpreted to verify the automated algorithm and to set its tolerances. For nLC retention time reproducibility, 95% of the 497 species had a 95% confidence interval of the mean of [+ or -] 0.9 min or less without the use of chromatographic alignment procedures. Furthermore, 95% of the 497 species had a mass measurement precision of [less than or equal to] 3.2 and [less than or equal to] 6.3 ppm for internally and externally calibrated spectra, respectively. Moreover, 95% of replicate ion abundance measurements, covering an ion abundance range of ~3 orders of magnitude, had a coefficient of variation of less than 62% without using any normalization functions. The variability of ion abundance was independent of LC retention time, mass, and ion abundance quartile. These measures of analytical reproducibility establish a statistical rationale for differentiating healthy and disease patient populations for the elucidation of biomarkers in the low molecular fraction of serum.

Byline: R. S. Minvaleev (1), A. D. Nozdrachev (1), V. V. Kir'yanova (2), A. I. Ivanov (1) Abstract: Cortisol, testosterone, dehydroepiandrosterone (DHEA), and aldosterone were measured in the blood serum of healthy subjects who adopted the cobra posture (bhujangasana), a key posture of hatha yoga. The subjects were trained in yoga blood samples were taken before and after the exercise at an interval of no more than 5 min. As revealed with a new confidence interval--evaluating method developed by A.I. Ivanov, cortisol decreased by 11% with a reliability of 0.99 in all subjects, while testosterone increased by 16% with a reliability of 0.95. Changes in DHEA and aldosterone varied. Proceeding from ideas about motor--visceral interactions, the authors hypothesize that, when the subjects adopted the cobra posture, the production of steroid hormones was controlled by neural mechanisms. Author Affiliation: (1) St. Petersburg State University, St. Petersburg, 199164, Russia (2) St. Petersburg Medical Academy of Postgraduate Studies, St. Petersburg, Russia Article History: Registration Date: 09/10/2004

Byline: Rade B. Vukmir (1) Keywords: serum prolactin; seizure; emergency Abstract: Abstract. Study objective: We sought to evaluate whether there was a correlation between elevated serum prolactin in patients presenting with the question of seizure. Methods: A Convenience sample of 200 consecutive patients were chosen who had a serum prolactin measurement in the setting of seizure activity. Results: The prolactin level was within a range of 3.90--294.00 mg/dl with an upper limit of normal being 29.9 mg/dl. Patients were ultimately diagnosed with seizure in 54.5% (109 of 200) with an abnormal prolactine in 31.0% (62 of 200). The sensitivity of this serum prolactin was 42%, the specificity was 82%, positive predictive value (PPV) of 74%, and negative predictive value (NPV) of 54%. There was an overall accuracy of 60% in the diagnosis of seizure, with a likelihood ratio of 2.4 (95% Confidence Interval [CI], 1.5--3.9). Conclusion: The measurement of serum prolactin is helpful as a confirmatory test, but not as screening test in the emergency department setting. Author Affiliation: (1) University of Pittsburgh, Medical Center UPMC, Northwest One Spruce Street, Franklin, PA, 16323, USA Article History: Registration Date: 01/01/2004 Received Date: 14/08/2003 Accepted Date: 30/01/2004

Byline: A. A. Kuznetsova (1), Yu. V. Natochin (2) Abstract: The blood serum osmolality and ion concentrations were studied in 144 children (healthy or having some diseases), and no deviations from the universally accepted normal values were found. Although the mean values were in the normal range, the diseases were associated with an increased variation of serum osmolality and ion concentrations, indicating an imbalance of the regulatory system. Normally, human urine is hyperosmolal and the urination depends on the excretion of osmotically active substances. This determines a physiological paradox: an increase in diuresis is accompanied by a concurrent increase in reabsorption of osmotically free water. Author Affiliation: (1) St. Petersburg State Pediatric Medical Academy, St. Petersburg, 194000, Russia (2) Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersburg, 194223, Russia Article History: Registration Date: 06/10/2004

Byline: Rana Karabudak (1), AslA+- Kurne (1), Dicle Guc (2), Meltem Sengelen (3), Hande Canpinar (2), Emin Kansu (2) Keywords: multiple sclerosis; IFN [beta]1a; matrix metalloproteinases; tissue inhibitors of MMPs; MMP-9; TIMP-1 Abstract: Abstract. There is emerging evidence that matrix metalloproteinases (MMPs) might be involved in blood brain barrier (BBB) breakdown in multiple sclerosis. A group of natural tissue inhibitors of metalloproteinases (TIMPS) regulates proteolytic activity to prevent tissue damage. TIMP-1 and MMP-9 are known to be secreted as heterodimers and TIMP-1 preferentially functions to inhibit MMP-9 activity. In this present study, the effects of IFN[beta]-1a on serum MMP-9 and TIMP-1 were evaluated longitudinally during a one-year period. The MMP-9 levels showed no significant changes while TIMP-1 levels gradually and significantly increased during 3.sup.rd and 6.sup.th months of therapy compared with pretreatment levels. Author Affiliation: (1) Hacettepe University, Faculty of Medicine, Department of Neurology Neuroimmunology Unit, 06100, Ankara, Turkey (2) Hacettepe University, Oncology Institute, Department of Basic Oncology, Ankara, Turkey (3) Hacettepe University, Oncology Institute, Department of Preventive Oncology, Ankara, Turkey Article History: Registration Date: 01/01/2004 Received Date: 05/11/2002 Accepted Date: 24/09/2003

Byline: S. Minisola (1), S. Dionisi (1), M. T. Pacitti (1), F. Paglia (1), V. Carnevale (2), A. Scillitani (3), S. Mazzaferro (1), S. De Geronimo (1), J. Pepe (1), E. D'Erasmo (1), E. Romagnoli (4) Keywords: Key words:Aging -- Bone disease -- Bone resorption -- Gender differences -- Serum markers Abstract: To assess how two different serum markers of bone resorption may reflect changes in bone turnover, we compared age- and sex-related changes in serum C-terminal telopeptide of type I collagen ([beta]CTx) and tartrate-resistant acid phosphatase activity (TRAP) in 136 healthy men and 184 normal women. Serum levels of the two markers were also assessed in several groups of patients of both sexes presenting with the most common metabolic and endocrine bone diseases: established osteoporosis (n= 77), primary hyperparathyroidism (n= 44), glucocorticoid excess (n= 17), chronic renal failure (n= 39), active Paget's disease of bone (n= 5), humoral hypercalcemia of malignancy (na=a3), osteomalacia (n= 3), hyperthyroidism (n= 10), post-surgical hypoparathyroidism (n= 10), acromegaly (active disease, n= 8) and Cushing's syndrome (n= 10). In men the regression of [beta]CTx with age showed an initial decrease in bone resorption followed by an increase thereafter, starting from the sixth decade of life. No age-related change in serum TRAP activity was observed. In women, by contrast, a slight but significant linear correlation of both serum [beta]CTx and TRAP with age (r= 0.223, p&lt 0.003 and r= 0.333, p&lt 0.0001, respectively) was found, the two markers being positively correlated (r= 0.238, p&lt 0.002). In each class of patients the mean Z-scores of [beta]CTx were significantly higher than those of TRAP activity. Moreover, compared with normal subjects, serum [beta]CTx seems to be characterized by a superior sensitivity relative to TRAP measurement, at least in the disorders studied. Author Affiliation: (1) Department of Clinical Sciences, University of Rome 'La Sapienza', IT (2) Department of Internal Medicine, IT (3) Department of Endocrinology, 'Casa Sollievo della Sofferenza' Hospital, IRCCS, San Giovanni Rotondo , IT (4) San Giovanni Battista Hospital, Rome, Italy, IT Article note: Received: 14 February 2001 / Accepted: 3 September 2001

Byline: U. Lange (1), O. Jung (2), J. Teichmann (3), G. Neeck (1) Keywords: Key words:Ankylosing spondylitis -- Disease activity -- Osteopenia -- Osteoporosis -- Vitamin D metabolism Abstract: Vertebral fractures due to osteoporosis are a common but frequently unrecognized complication of ankylosing spondylitis (AS) and various factors may contribute to the development of osteoporosis in AS. It is known that inflammatory activity in rheumatic disease (i.e., proinflammatory cytokines) itself plays a possible role in the pathophysiology of bone loss. 1,25-Dihydroxyvitamin D.sub.3 (1,25(O[H).sub.2]D.sub.3) seems to be another possible candidate for mediatory function in regulating both the inflammatory process and bone turnover. The aim of this study was to evaluate the relation between disease activity, bone turnover and calciotropic hormones. In 70 patients with established AS and an age- and sex-matched control group, the relation between disease activity (erythrocyte sedimentation rate, C-reactive protein, Bath Ankylosing Spondylitis Disease Activity Index), and serum levels of vitamin D metabolites, parathyroid hormone (PTH), bone alkaline phosphatase (bAP) and urinary pyridinium crosslinks were determined. Serum levels of 1,25(O[H).sub.2]D.sub.3 (p&lt 0.01) and PTH (p&lt 0.01) were negatively correlated with disease activity, the excretion of urinary pyridinium crosslinks showed a positive correlation with disease activity (p&lt 0.01), and 1,25(O[H).sub.2]D.sub.3 and PTH were positively correlated with bAP (p&lt 0.01). These results indicate that high disease activity in AS is associated with an alteration in vitamin D metabolism and increased bone resorption. Furthermore, the decreased levels of 1,25(O[H).sub.2]D.sub.3 may contribute to a negative calcium balance and inhibition of bone formation. Our results suggest further research is necessary to determine whether low levels of 1,25(O[H).sub.2]D.sub.3 as an endogenous immune modulator suppressing activated T cells and cell proliferation may accelerate the inflammation process in AS. Author Affiliation: (1) Kerckhoff Clinic and Foundation, Department of Rheumatology, University of Giessen, Bad Nauheim, DE (2) IV. Medical Clinic -- Nephrology, University of Frankfurt , DE (3) III. Medical Clinic, University of Giessen, Germany, DE Article note: Received: 29 January 2001 / Accepted: 3 August 2001

Apratoxin A is considered as a potent novel cytotoxin from the marine cyanobacterium Lyngbya majuscula and its structure is discussed.

Research has been conducted on the fortified sera. The comparison of these sera to the fetal bovine serum has been performed in investigating their ability to increase kidney cell sensitivity to viral infections.

Research has been conducted on the dopamine analogue 6-hydroxydopamine (6-OHDA). The interaction of 6-OHDA with serum ferric transferrin has been examined for toxicity.

Thirty-six primiparous heifers were used to determine the effect of dietary energy on postpartum interval, milk production, serum concentrations of insulin, insulin-like growth factor I (IGF-I), growth hormone, and cerebrospinal fluid (CSF) concentrations of neuropeptide-Y (NPY). Low-quality hay was fed during the last trimester of pregnancy to achieve suboptimal calving weight (370 +/- 5 kg) and condition score (4.0 +/- .1). After calving, cows were allotted to one of four dietary treatments that differed in metabolizable energy. Experimental diets were fed at 2.5% of shrunk body weight and formulated to provide 1.8 (low), 2.1 (maintenance), 2.4 (maintenance high), or 2.7 (high) Mcal of ME/kg DM. Daily milk production was estimated at approximately 30, 60, and 90 d postpartum. Condition score change and weight change were defined as change from calving to d 90 postpartum. As energy intake increased, condition score change (P < .001), IGFI (P < .001) and insulin (P < .01) increased and postpartum interval decreased (P = .04). No relationship existed between postpartum interval and CSF concentration of NPY (P > .1). Condition score change was positively associated with IGF-I and insulin (r = .71, P < .001; r = .38, P = .02, respectively) and negatively associated with GH (r = -.67, P < .001). Weight change and serum concentrations of GH did not differ (P > .10) among treatments. Increasing dietary energy intake was associated with a curvilinear increase in milk yield (P = .04) and percentage milk fat (P = .03) and a linear increase (P = .04) in energy available for milk production. Greater milk yield at d 30 was associated with a longer postpartum interval (r = .34, P < .05). In conclusion, a greater proportion of net energy is partitioned to maternal tissue realimentation when cows receive high compared to low-, maintenance-, or moderate-energy diets. Key Words: Beef Cows, Energy Balance, Milk Production, Postpartum Interval

Research has been conducted on two exchanger isoforms Na (super)+ /H (super)+, NHE1 and NHE3. The identification of the domains that are involved in the NHE3 targeting has been investigated.

Research has been conducted on the iron-binding properties of the N-lobe following ligand mutations. The changes resulting from mutating His 249 are discussed.

The recombinant serum amyloid A protein (SAA) effects on high-density lipoprotein (HDL) have been investigated to determine whether pre beta 1-HDL is generated by SAA from alpha-migrating HDL. Results indicate the possibility that SAA-mediated HDL remodeling takes place in inflammation.

A study concerning nitrating intermediates that act as physiological catalysts for the lipid peroxidation initiation and oxidized lipids and sterols formation is presented. Results indicate that phagocytes can employ myeloperoxidase-generated reactive nitrogen intermediates.

Research has been conducted on the alpha particle emitters. The design of ionophores derived from calix[4]arene to have selectivity for Ra (super)2+ over the lighter alkaline earth metal ions is described.

Research has been conducted on the improvement in the selective targeting of compound gemcitabine and other cytotoxic agents to solid tumors. Results demonstrate that these improvements can be enhanced by conjugation to ligands targeting peripheral benzodiazepine receptors.

Research was conducted to examine the importance to human butyrylcholinesterase (BChE) tetramerization of seven selected conserved C-terminal aromatic residues, namely, Trp 543, Phe 547, Trp 550, Tyr 553, Trp 557, Phe 5661 and Tyr 564. Wild-type BChE C-terminal peptides interacted with one another independently from the remainder of the molecule in the yeast two-hybrid system. Results indicate that the conserved aromatic residues of the C-terminus are responsible for the stabilization of BChE tetramers and mediate the interactions to increase tetramers of BChE in the presence of poly-L-proline and the proline-rich attachment domain.

Research has been conducted on Candida albicans induced by serum. Results indicate that serum albumin does not affect C.albicans morphological change from the yeast cell form to hyphae.

Tetracycline has been determined in human serum samples by a combination of: (1) sychronous fluorescence spectra of whole sera treated with Mg(super 2+), and (2) the multivariate calibration methods of partial least-squares (PLS-1) and a variant of the recently introduced hybrid linear analysis (HLA), which does not require the knowledge of pure-component spectra. The calibration set was designed with 50 sera spiked with Tetracycline has been determined in human serum samples by a combination of: (1) concentrations of tetracycline in the range 0.0-4.0 mu g mL(super 1). Studies concerning validation, precision, accuracy and figures of merit (selectivity, sensitivity and limit of determination) were also carried out. A novel wavelength-selection procedure was applied to minimize the effect of nonmodeled interferents present in serum samples containing bilirubin, triglycerides, and salicylate. Overall, the performance of the newly developed HLA approach seems to be better than that of PLS-1.

The technique of automated in-tube solid phase microextraction (SPME) coupled with liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) was evaluated for the determination of beta-blockers in urine and serum samples. In-tube SPME is an extraction technique for organic compounds in aqueous samples, in which analytes are extracted from the sample directly into an open tubular capillary by repeated draw/eject cycles of sample solution. LC/MS analyses of beta-blockers were initially performed by liquid injection onto a LC column. Nine beta-blockers tested in this study gave very simple ESI mass spectra, and strong signals corresponding to [M + H](super +) were observed for all beta-blockers. The beta-blockers were separated with a Hypersil BDS C(sub 18) column using acetonitrile/methanol/water/acetic acid (15:15:70:1) as a mobile phase. To optimize the extraction of beta-blockers, several in-tube SPME parameters were examined. The optimum extraction conditions were 15 draw/eject cycles of 30 mu L of sample in 100 mM Tris-MC1 (pH 8.5) at a flow rate of 100 mu L/min using an Omegawax 250 capillary (Supelco, Bellefonte, PA). The beta-blockers extracted by the capillary were easily desorbed by mobile-phase flow, and carryover of beta-blockers was not observed. Using in-tube SPME/LC/ESI-MS with selected ion monitoring, the calibration curves of beta-blockers were linear in the range from 2 to 100 ng/mL with correlation coefficients above 0.9982 (n = 18) and detection limits (S/N = 3) of 0.1-1.2 ng/mL. This method was successfully applied to the analysis of biological samples without interference peaks. The recoveries of beta-blockers spiked into human urine and serum samples were above 84 and 71%, respectively. A serum sample from a patient administrated propranolol was analyzed using this method and both propranolol and its metabolites were detected.

Methyl 5-amino-1-(chloromethyl)-3-((5,6,7-trimethoxyindol-2-yl)carbonyl)-1,3-dihydr o-3H-pyrrolo(3,2-e)indole-7-carboxylate, an amino analogue of the anticancer antibiotic and potent DNA minor groove alkylating agent seco-duocarmycin SA, was synthesized. Important points in the synthesis are the sequential radical cyclization and Hemetsberger reaction steps to construct the indoline and indole rings of the target compound from a 1,2,3-trisubstituted benzene precursor.

Research was conducted to examine the pathogenicity of Helicobacter pylori with focus on the influence of VacA. A decrease in trans-epithelial resistance across the cell monolayer resulted from the adherence of H. pylori to Madin-Darby canine kidney cells. Results indicate the possibility that increasing the paracellular permeability of the gastric epithelium may be the specific role of VacA in ulcerogenesis.

The objectives of this study were to examine differences in scrotal circumference, sperm motility, and percentage of normal sperm cells between two lines of Angus beef cattle divergently selected for blood serum IGF-I concentration. Data were obtained from an ongoing experiment involving 100 spring-calving (50 high and 50 low line) and 100 fall-calving (50 high and 50 low line) purebred Angus cows. Scrotal circumference, percentage of motile sperm cells, and percentage of normal sperm cells did not differ between high and low IGF-I line yearling bulls (P = .79, .50, and .56, respectively). The IGF-I concentrations measured at d 28, 42, and 56 of the postweaning test are abbreviated as IGF28, IGF42, and IGF56, respectively. Coefficients for the quadratic regression of scrotal circumference on IGF28 and IGF42 tended to be negative (P = .07 and .08, respectively), as did the coefficient for the quadratic regression of the percentage of motile sperm cells on IGF42 (P = .08). The coefficient for the linear regression of percentage of normal sperm cells on IGF28 was positive (P = .02). The coefficient for the quadratic regression of percentage of normal sperm cells on IGF56 was negative (P = .04). Coefficients for the quadratic regression of scrotal circumference and percentage of normal sperm cells on mean IGF-I concentrations were negative and important (P = .04 and .08, respectively). Thus, scrotal circumference and percentage of normal sperm cells are related to blood serum IGF-I concentration in yearling Angus bulls. Key Words: Beef Cattle, Insulin-Like Growth Factor, Selection, Scrotum, Spermatozoa

Research was conducted to examine the synthesis of unit A of the cryptophycins from (s)-trans-3-penten-2-ol and from (s)-trans-4-hexen-3-ol using a (2,3)-Wittig rearrangement of a propargyl ether as the key stereodetermining step. The investigation involved the elaboration of the rearrangement product, selective ozonolytic cleavage and Wittig olefination. Results indicate that the introduction of the phenyl group at the end of the sequence provides a convenient access to cryptophycin analogues.