Alpha 1-acid Glycoprotein Concentration Research Articles

Urinary monocyte chemoattractant protein 1 and alpha 1 acid glycoprotein as biomarkers of renal disease activity in juvenile-onset systemic lupus erythematosus

A higher proportion of patients with juvenile-onset systemic lupus erythematosus (JSLE) will have renal involvement compared with adult-onset disease, some progressing to renal failure in adulthood. Histological examination is the gold standard for diagnosing lupus nephritis (LN), but its invasive nature limits routine use. Using cross-sectional cohort analysis, we aimed to determine whether urinary concentrations of monocyte chemoattractant protein-1 (MCP1), alpha-1-acid glycoprotein (AGP) and interferon-inducible protein 10 (IP10) are biomarkers of active LN. Sixty JSLE patients recruited to the UK JSLE Cohort Study were categorized according to the British Isles Lupus Assessment Group (BILAG) activity index. Patients with active renal JSLE (n = 8; renal BILAG score A, B) had significantly higher urinary MCP1 concentrations than patients with inactive renal disease (n = 52; renal BILAG score C, D, E; 582 pg/mg creatinine [Cr], 207 pg/mg Cr; p = 0.018) or healthy controls (n = 23; 117 pg/mg Cr; p = 0.005). Urinary AGP concentration was significantly elevated in patients with active renal disease compared with inactive renal disease (1517 ng/mg Cr, 485 ng/mg Cr; p = 0.027) or healthy controls (313 ng/mg Cr; p = 0.013). Urinary IP10 concentration was not significantly different between groups, but did strongly correlate with uMCP and uAGP levels (rho = 0.38, p = 0.009; rho = 0.33, p = 0.021). Urinary MCP1 and AGP are biomarkers of LN, providing insight into its pathophysiology. Longitudinal studies are warranted.

Effect of altered AGP plasma binding on heart rate changes by S(−)‐propranolol in rats using mechanism‐based estimations of in vivo receptor affinity (KB,vivo)

In contrast to the impact of plasma protein binding on pharmacokinetics, no quantitative in vivo information is available on its impact on pharmacodynamics. The pharmacokinetic–pharmacodynamic relationship of the model drug S(−)‐propranolol was evaluated using mechanism‐based estimations of in vivo receptor affinity (KB,vivo), under conditions of altered plasma protein binding resulting from different levels of alpha‐1‐acid glycoprotein (AGP). Male Wistar Kyoto rats with isoprenaline‐induced tachycardia received an intravenous infusion of S(−)‐propranolol, on postsurgery day 2 (n = 7) and day 7 (n = 8) with elevated and normal plasma protein binding, respectively. Serial blood samples were taken in parallel to heart rate measurements. AGP concentrations at 2 and 7 days postsurgery were 708 ± 274 and 176 ± 111 µg/mL (mean ± SE), respectively. Using nonlinear mixed effects modeling, AGP concentration was a covariate for intercompartmental clearance for the third compartment of the pharmacokinetic model of S(−)‐propranolol. Individual values of AGP concentrations ranged between 110 and 1150 µg/mL, and were associated with KB,vivo values of S(−)‐propranolol from 7.0 to 30 nM. Using the KB,vivo for S(−)‐propranolol with correction for average values for normal and elevated plasma protein binding, nearly identical values were found. This confirms, strictly quantitative, earlier indications that plasma protein binding restricts the pharmacodynamics of S(−)‐propranolol. © 2009 Wiley‐Liss, Inc. and the American Pharmacists Association J Pharm Sci 99: 2511–2520, 2010

Serum α‐1‐acid glycoprotein concentration in clinically healthy puppies and adult dogs and in dogs with various diseases

alpha-1-acid glycoprotein (AGP) is an acute-phase protein and a serum marker of inflammation and neoplasia in humans. AGP concentrations in diseased dogs and the potential effects of age, breed, and sex have not been elucidated. The purpose of this study was to examine differences in AGP concentration based on age, sex, and breed in a large population of clinically healthy dogs and to compare AGP concentrations in dogs with various diseases. Serum was obtained from clinically healthy puppies (n=74) and adults (n=172) of both sexes, and included mongrels (n=205) and Beagles (n=41). Serum also was obtained from 192 dogs with various diseases, including 8 with pyometra that were sampled before, and 1, 2, 3, and 10 days after surgery. AGP concentration was measured by single radial immunodiffusion. Statistical comparisons were made among age, sex, breed, and disease groups. Serum AGP in healthy adult mongrels was 364+/-106 mg/L (reference interval, 152-576 mg/L). AGP was lowest in newborns (n=11, 122+/-54 mg/L) and gradually increased to adult levels by 3 months of age. Median AGP concentration was highest in dogs with parvovirus (n=17, 2100 mg/L), distemper (n=7, 1250 mg/L), and pyometra (n=18, 2480 mg/L) and was also significantly higher in dogs with acute filariasis, renal failure, urolithiasis, pancreatitis, hepatitis, trauma, hyperadrenocorticism, and immune-mediated hemolytic anemia. Dogs with acute filariasis and acute hepatopathy had significantly higher AGP concentrations than dogs with chronic filariasis and chronic hepatopathy. Serum AGP concentration decreased gradually following surgery for pyometra but remained increased after 10 days (896+/-175 mg/L). Because of significantly lower AGP in puppies, the age of dogs should be considered when using AGP as a marker of disease. Serum AGP may be a useful marker of inflammatory disease in dogs and may help differentiate acute and chronic stages of disease.

Lopinavir protein binding in HIV‐1‐infected pregnant women

Pregnancy may alter protein binding (PB) of highly bound protease inhibitors due to changes in plasma concentrations of albumin and alpha-1 acid glycoprotein (AAG). Small changes in PB can greatly impact the fraction of drug unbound (FU) exerting pharmacological effect. We report lopinavir (LPV) PB during third trimester (antepartum, AP) compared to > or =1.7 weeks postpartum (PP) to determine if FU changes compensate for reduced total concentrations reported previously. P1026s enrolled women receiving LPV/ritonavir, soft gel capsules 400/100 mg or 533/133 mg twice daily. LPV FU, albumin and AAG were determined AP and PP. AP/PP samples were available from 29/25 women respectively with all but one woman receiving the same dose AP/PP. LPV FU was increased 18% AP vs. PP (mean 0.96+/-0.16% AP vs. 0.82+/-0.21% PP, P=0.001). Mean protein concentrations were reduced AP (AAG=477 mg/L; albumin=3.28 mg/dL) vs. PP (AAG=1007 mg/L; albumin=3.85 mg/dL) (P<0.0001 for each comparison). AAG concentration correlated with LPV binding. Total LPV concentration did not correlate with LPV FU AP or PP. However, higher LPV concentration PP was associated with reduced PB and higher FU after adjustment for AAG. LPV FU was higher and AAG lower AP vs. PP. The 18% increase in LPV FU AP is smaller than the reduction in total LPV concentration reported previously and is not of sufficient magnitude to eliminate the need for an increased dose during pregnancy.

Serum protein biomarkers screening in HCC patients with liver cirrhosis by ICAT-LC-MS/MS

The prognosis of HCC remains poor mainly because of the lack of diagnosis biomarkers especially in patients with cirrhosis background. To identify serum biomarkers for HCC, we use cleavable stable isotope labeling (cICAT) combined with LC-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) to compare the serum proteome between liver cirrhosis and HCC patients. Sera from nine liver cirrhosis patients and nine HCC patients were selected for screening study. Then the alpha-1-acid glycoprotein (AGP) was chosen for validation by western blot. AGP was measured in the separate validation study including 52 HCC patients and 40 liver cirrhosis patients by rate nephelometry. Its value for HCC diagnosis from liver cirrhosis was also appreciated through receiver operating curve (ROC). We quantified and identified 31 different proteins which include AGP, complement C4, haptoglobin, alpha-1-antitrypsin precursor, alpha-2-macroglobulin precursor, prothrombin precursor and ubiquitin carboxyl-terminal hydrolase 44 (USP44), etc. The serum concentration of AGP in HCC patients (n = 52) was significantly higher than in liver cirrhosis (n = 40) (P < 0.005) in validation study. AGP was useful for discrimination of the HCC cases from LC patients when the AFP levels were below 500 ng/ml. The area under curve (AUC) of the AGP and the combination of AGP and AFP were 0.834 (P < 0.0005, 95% CI: 0.734-0.934) and 0.880 (P < 0.0005, 95% CI, 0.789-0.970) separately, which are higher than AFP alone (0.538, P = 0.604, 95% CI: 0.386-0.689). cICAT combined with LC-MS/MS-based serum proteome analysis can be useful in the screening of serum biomarkers for HCC. Alpha-1-acid glycoprotein combining AFP could aid the diagnosis of HCC.

Relationship between serum acute-phase proteins and high disease activity in patients with rheumatoid arthritis

The aim of this study was to assess the relationship between serum acute-phase proteins and high disease activity evaluated by activity score (DAS28) in patients with rheumatoid arthritis. Studies were carried out on 27 females with RA and 32 control women. Acute-phase proteins were divided into 4 fractions as follows: alpha1-globulins represented by alpha1-acid glycoprotein (AGP) and alpha1-antitrypsin (AAT); alpha2-globulins - haptoglobin (Hp); beta-globulins - complement C3 (C3) and total transferrin (Tf); gamma-globulins - C reactive protein (CRP), rheumatoid factor (RF) and immunoglobulin G (IgG), and determined by immunoturbidimetric methods. The serum levels of acute-phase proteins changed in RA patients. On account of the alterations of concentration, acute-phase proteins are placed in the downgrade scale as follows: CRP, Hp, AGP, C3, AAT and Tf. None of the acute-phase proteins correlated with the RF and the majority of them were closely related to ESR. Almost all of the acute-phase proteins (without C3) were closely related to RA activity (based on DAS28) and their places in the downgrade scale were as follows: CRP, Tf, AGP, Hp and AAT. The degree of disability evaluated by Health Assessment Questionnaire has affected on the concentrations of AGP, Tf and CRP. Serum AGP, AAT and RF levels significantly correlated with the patient's age. No correlations were observed between IgG, TP levels, and clinical data. Among the entire panel, the CRP and AGP appeared to be the most useful biochemical markers for evaluation of the disease activity of patients with RA.

Effect of a high saturated fatty acids load on serum concentrations of C-reactive protein, alpha1-antitrypsin, fibrinogen and alpha1-acid glycoprotein in obese women

Obesity is associated with increased inflammation. Creactive protein (CRP) and inflammation-sensitive plasma protein (ISPs) are inflammatory markers. Proinflammatory process may be influenced by high saturated fatty acid intake. The aim of the present study was to evaluate the role of saturated fatty acids load on postprandial circulating levels of PCR and ISPs (alpha1-antitrypsin, alpha1-acid glucoprotein, and fibrinogen) in obese women. A total of 15 obese women (age = 31,7 +/- 4,5 years, BMI = 37,9 +/- 7,3 kg/m(2)) and 15 lean controls women (age = 30,6 +/- 4,6 years, BMI = 20,6 +/- 2,6 kg/m(2)) were recruited for this study. After and overnight fast subjects ate the fat load consisted of 75 g of fat (100% saturated fatty acid, 0% cholesterol), 5 g of carbohydrates, and 6 g of protein per m2 body surface area. Postprandial serum levels of CRP, alpha1-antitrypsin, alpha1-acid glucoprotein, and fibrinogen were measured. Anthropometry and blood biochemical parameters were measured in both groups. The obese women had fasting serum PCR levels higher (p = 0,013) and fibrinogen (p = 0,04) than those of control women. Serum CRP and fibrinogen levels was positively related to body mass index (BMI) in obese group. There weren't differences in fasting serum alpha1- antitrypsin levels (p = 0,40), and alpha1-acid glucoprotein (p = 0,28) levels in obese group in comparison to lean control group. Serum CRP, alpha1-antitrypsin, alpha1-acid glucoprotein, and fibrinogen did not change postprandially (p = > 0,05 difference to fasting levels). A high-saturated fatty acids load is not associated with serum CRP, alpha1-antitrypsin, alpha1-acid glucoprotein, and fibrinogen levels increase. Serum alpha1-antitripsin and alpha1-acid glucoprotein levels are not increased in obese women. Serum PCR and fibrinogen levels are increased in obese women, and are positively related to BMI.

Traits associated with innate and adaptive immunity in pigs: heritability and associations with performance under different health status conditions

There is a need for genetic markers or biomarkers that can predict resistance towards a wide range of infectious diseases, especially within a health environment typical of commercial farms. Such markers also need to be heritable under these conditions and ideally correlate with commercial performance traits. In this study, we estimated the heritabilities of a wide range of immune traits, as potential biomarkers, and measured their relationship with performance within both specific pathogen-free (SPF) and non-SPF environments. Immune traits were measured in 674 SPF pigs and 606 non-SPF pigs, which were subsets of the populations for which we had performance measurements (average daily gain), viz. 1549 SPF pigs and 1093 non-SPF pigs. Immune traits measured included total and differential white blood cell counts, peripheral blood mononuclear leucocyte (PBML) subsets (CD4+ cells, total CD8α+ cells, classical CD8αβ+ cells, CD11R1+ cells (CD8α+ and CD8α-), B cells, monocytes and CD16+ cells) and acute phase proteins (alpha-1 acid glycoprotein (AGP), haptoglobin, C-reactive protein (CRP) and transthyretin). Nearly all traits tested were heritable regardless of health status, although the heritability estimate for average daily gain was lower under non-SPF conditions. There were also negative genetic correlations between performance and the following immune traits: CD11R1+ cells, monocytes and the acute phase protein AGP. The strength of the association between performance and AGP was not affected by health status. However, negative genetic correlations were only apparent between performance and monocytes under SPF conditions and between performance and CD11R1+ cells under non-SPF conditions. Although we cannot infer causality in these relationships, these results suggest a role for using some immune traits, particularly CD11R1+ cells or AGP concentrations, as predictors of pig performance under the lower health status conditions associated with commercial farms.

Results From a Randomized Trial of Salvage Chemotherapy Followed by Lestaurtinib for FLT3 Mutant AML Patients in First Relapse.

Abstract 788FLT3 mutations are common in AML and are associated with poor prognosis. Lestaurtinib is a multi-targeted kinase inhibitor with potent activity against FLT3. In an attempt to definitively establish whether or not there is clinical benefit for FLT3 inhibition in the relapse setting, we conducted a randomized trial of chemotherapy followed by lestaurtinib for patients with FLT3 mutant AML in first relapse. Patients were randomized to receive either chemotherapy alone, consisting of mitoxantrone, etoposide, and cytarabine (MEC) or high-dose cytarabine (HiDAc) depending on the duration of CR1) or chemotherapy followed by 80 mg lestaurtinib orally BID. Patients on the chemotherapy only arm were eligible for crossover if they were refractory to therapy. Patients on the lestaurtinib arm received the drug for up to 112 days on study (they could continue to receive the drug through a separate extension protocol). The primary objective was complete remission (CR/CRp). Secondary objectives included overall survival (OS), safety, and tolerability. From January 2004 through December 2008, 224 patients at 52 different centers were enrolled and randomized, with 220 patients actually receiving therapy. Blood samples from patients on the lestaurtinib arm were obtained at baseline, on Day 15, and on Day 42 for pK studies and for determination of in vivo FLT3 inhibition and measurement of plasma FLT3 ligand (FL) and alpha-1 acid glycoprotein (AGP) levels. The median age for all patients enrolled was 55 years (53.5 in the control arm, 58.5 in the lestaurtinib arm), with 88% having FLT3/ITD mutations, 8% having D835 mutations, and 4% having both. The duration of CR1 was less than 6 months in 47% of patients in each arm. On the lestaurtinib arm, 107 patients received at least 1 week of lestaurtinib (mean 60.1 days, range 8-418), with 17 of these patients receiving drug on the extension protocol. On the chemotherapy only arm, 7 patients crossed over to receive lestaurtinib, and 31 patients went on to receive lestaurtinib on the extension protocol after outcome evaluation. Lestaurtinib was generally well tolerated following chemotherapy, with 104 Grade 3/4 adverse events in the lestaurtinib arm versus 99 in the control arm. There were 61 serious adverse events in the lestaurtinib arm versus 49 in the control arm. Early death (by Day 42) occurred in 22 lestaurtinib patients (2 progressive leukemia, 14 infectious, 6 organ failure) and16 control patients (6 progressive leukemia, 5 infectious, 5 organ failure). By intention to treat analysis, there were 29 CR/CRp in the lestaurtinib arm and 23 in the control arm (26% versus 21%; p = 0.35), with no difference in OS (4.73 months versus 4.57 months) between the two arms. Blood samples from 79 of the 107 lestaurtinib-treated patients obtained on Day 15 were analyzed for FLT3 inhibitory activity (plasma inhibitory activity [PIA] assay), as well as FLT3 ligand (FL), lestaurtinib, and AGP concentrations. Mean lestaurtinib plasma concentration was 12 uM at Day 15 and fell to 7.5 uM on Day 42. Mean lestaurtinib concentration at Day 15 was 13.3 uM in patients achieving CR/CRp and 11.5uM in non-responders. A target of >85% inhibition of FLT3 maintained at trough (pre-dose) was defined from previous studies. Of the 79 patients tested, 46 (58%) achieved this degree of FLT3 inhibition on Day 15. FL concentrations rose from baseline (15.6 pg/mL) to Day 15 (1148 pg/mL), and AGP concentrations rose by an average of 52% over the same period, both of which may have reduced the degree of FLT3 inhibition. Of the 46 patients with target FLT3 inhibition on Day 15, 18 (39%) achieved CR/CRp, while only 3 (9%) of the 32 patients with below target FLT3 inhibition achieved CR/CRp. Thus, in patients with FLT3 mutant AML at first relapse, pharmacokinetic factors and possible physiologic factors (FL and AGP) limit lestaurtinib's ability to effectively inhibit FLT3. FLT3 inhibition by lestaurtinib, when achieved, correlates with better CR rates, but in this trial that benefit was negated by a poor CR rate in those patients for whom the drug did not reach the target level of FLT3 inhibition. Overall, lestaurtinib treatment following re-induction chemotherapy failed to increase response rates or prolong survival of patients with FLT3 mutant AML in first relapse. Disclosures:Levis:Cephalon: Clinical Advisory Board member. Ravandi:Cephalon: Honoraria, Member, clinical advisory board, Research Funding. Erba:Cephalon: Research Funding, Speakers Bureau. Baccarani:Novartis Pharma, Bristol Myers Squibb, Merck Sharp & Dome, Pfizer: Consultancy, Speakers Bureau. Stone:Cephalon: ad hoc consultancy; Novartis: Research Funding, ad hoc consultancy. Advani:Cephalon: Research Funding. Douer:Cephalon: Honoraria, Research Funding, Speakers Bureau. Litzow:Cephalon: Research Funding. Tremmel:Cephalon: Employment, Equity Ownership. Bensen-Kennedy:cephalon: Employment. Smith:Cephalon: Research Funding.

Serum α1-Acid Glycoprotein Level Associate with Imatinib Trough Plasma Concentration and Clinical Outcome in CML Patients Treated with Imatinib: Blood Level Testing of Imatinib in Nagoya (BINGO) Study.

Abstract 4266 IntroductionImatinib, which is the most successful molecularly targeting drug, is the first-line agent for the treatment of chronic myeloid leukemia patients (CML) in chronic phase (CP). However, some patients show poor response against imatinib therapy, resulting in the progression to blastic phase. To date, several reports showed that imatinib trough plasma concentration (Cmin) was associated with the clinical response in CML patients, while it remains unclear whether Cmin reflects the actual inhibitory effect on the BCR-ABL kinase activity. In human plasma, imatinib binds to several plasma proteins, such as albumin (ALB) and alpha1-acid glycoprotein (AGP), and the protein-binding-imatinib lacks the kinase inhibitory activity. Especially, AGP is a major imatinib-binding protein, and the high plasma AGP concentration reportedly reduces the clinical efficacy of imatinib. These results indicate that the free-imatinib concentration is a more reliable surrogate marker for predicting the clinical efficacy than the total imatinib concentration. However, since routine measurements of the free-imatinib concentration is difficult, the inhibitory activity of the patient's plasma (PIA) against the BCR-ABL kinase is proposed to be a direct marker reflecting the actual inhibitory effects of the free-imatinib. In this study, we analyzed the association between Cmin of total imatinib and the clinical response in CML-CP patients in considering the AGP concentration. Furthermore, we evaluate the usefulness of PIA for predicting the clinical response in the CML patients treated with imatinib. MethodsWe measured Cmin of the total imatinib and AGP and ALB concentrations at the different two points in CML-CP patients who were treated with imatinib alone. The PIA was quantified by determining the de-phosphorylation levels of BCR-ABL and STAT5 in BCR-ABL-expressing Ba/F3 cells after the treatment with patients' plasma. We evaluated the association of these parameters with clinical response. Informed consent was obtained from all patients to use their samples for this study. This study was approved by the ethical committee of Nagoya university school of medicine. ResultsThe study population included 65 CML-CP patients. The mean Cmin of imatinib was 1070 ± 362 ng/ml. Clinical response was evaluated by the achievement of the complete cytogenetic response (CCR) and major molecular response (MMR) at 12 months after the start of imatinib therapy. The CCR and MMR were achieved in 63 (96.9%) and 40 (61.5%) patients, respectively. The mean Cmin of the patients who achieved MMR was significantly higher than that of those who did not (1167 ± 386 ng/ml vs. 913 ± 257ng/ml, P=0.012). The differences of Cmin between the two different points (median interval 28 ± 20 days) in the same patient were not so little (median 123 ± 198 ng/ml, range 12 to 923 ng/ml), but were not associated with clinical outcomes. The PIA was statistically correlated with the Cmin (P=0.018). Although serum AGP level was within normal range (72.8 ± 13.9 mg/dl, range 44 to 106 mg/dl) in most patients, it was notable that the serum AGP level correlated with Cmin of imatinib and clinical outcomes. The Cmin of patients with high AGP level was significantly higher than that of low AGP level (1237 ± 379ng/ml vs. 907 ± 260ng/ml, P=0.0003) and the MMR rate was significantly higher in patients with high AGP level (78%) than in those with low AGP level (45%, P=0.007). ConclusionsWe demonstrate that the Cmin of imatinib predicts clinical response and reflects the actual kinase inhibitory effects in CML patients treated with imatinib. However, the Cmin is not necessarily stable during the treatment in a part of patients, suggesting that it is recommended to examine Cmin at more than two different points for evaluating its suitability. The serum AGP and ALB levels did not affect the relationship between Cmin and PIA. However, since the serum AGP level was correlated with Cmin and clinical response, the AGP level might influence the kinase inhibitory activity in the patients whose Cmin of imatinib were low. Disclosures:Kiyoi:Novartis Pharma : Research Funding; Kyowa Hakko Kirin Co., Ltd. : Consultancy. Naoe:Kyowa Hakko Kirin Co., Ltd. : Research Funding; Chugai Pharmaceutical Co.,Ltd.: Research Funding; Wyeth K.K.: Research Funding.

Enhanced expression of α1-acid glycoprotein and fucosylation in hepatitis B patients provides an insight into pathogenesis

Altered glycosylation and concentration of alpha1-acid glycoprotein has been known to be related to the pathogenesis of the hepatic diseases. The present study investigated enhanced fucosylation of AGP in the sera of chronic hepatitis B (HBV-CH) and hepatitis B cirrhosis (HBV-LC) patients by high performance anion exchange chromatography and by ELISA using fucose binding Aleuria aurantia lectin. The concentration of AGP determined by ELISA using monoclonal anti-human AGP (mAb-AGP) showed high level of AGP in HBV-CH and HBV-LC patients. This was further judged by association constant (K (A)) measured by surface plasmon resonance analysis. There was no apparent linkage variation of sialic acid among different patient groups when tested with two sialic acid binding lectins viz., Maackia amurensis agglutinin (MAA, NeuAc alpha2-3-) and Sambucus nigra agglutinin (SNA, NeuAc alpha2-6-) respectively. There was no change of oligosaccharide branching in HBV-CH in comparison to controls whereas a slight change was observed in HBV-LC using ConA. The above results suggest that the changes in concentration of AGP and fucosylation have a prognostic value of hepatitis diseases and it could be possible to use AGP as diagnostic marker besides clinical examination and routine laboratory investigation.

Serum Acute Phase Protein Concentrations in Dogs with Autoimmune Hemolytic Anemia

Serial monitoring of acute phase protein (APP) concentrations in canine autoimmune hemolytic anemia (AIHA) has not been reported. Acute canine AIHA is accompanied by an acute phase response (APR) characterized by increased C-reactive protein (CRP) and alpha1-acid glycoprotein (AAG) concentrations and decreased albumin concentrations. Twenty-seven dogs with AIHA and 11 control dogs. Prospective, cohort study. CRP, AAG, and albumin concentrations, white blood cell (WBC) count, and packed cell volume (PCV) were determined at admission (day 1), every 48 hours until death or discharge, and on days 30, 90, 180, and 365. Compared with controls, CRP and AAG concentrations were increased and albumin concentration was decreased in dogs with AIHA (days 1-7; P < .002) and normalized with disease stabilization (days 9-365; P > .05). APP concentrations on day 1 were not predictive of survival, duration of hospitalization, or number of blood transfusions (P= .153-.940). PCV correlated with APP concentrations over time (CRP r=-.600, AAG r=-.665, albumin r= .533; P < .0001) as did WBC count (CRP r= .253, AAG r= .486, albumin r=-.246; P < .006). Day 1 CRP concentration was lower for dogs that received corticosteroids before referral (115.3 microg/mL) compared with dogs that did not (191.2 microg/mL; P= .02). An APR occurs in canine AIHA. Initial APP concentrations are not predictive of acute survival, correlate with hematologic markers of remission, and normalize rapidly with disease stabilization.

α1-acid glycoprotein and α1-antitrypsin as early markers of treatment response in patients receiving the intensive phase of tuberculosis therapy

The identification of early markers that predict the response to anti-tuberculosis treatment would facilitate evaluation of new drugs and improve patient management. This study aimed to determine whether selected acute phase proteins and micronutrients measured at the time of diagnosis and during the first weeks of treatment could predict treatment responses during the 2-month standard intensive phase of therapy. For this purpose, alpha1-antitrypsin, alpha1-acid glycoprotein, alpha2-macroglobulin, C-reactive protein, C3, C4, zinc, copper and selenium concentrations were measured in Brazilian patients with smear-positive tuberculosis at the time of diagnosis and 1, 3, 5 and 8 weeks after initiation of therapy. Patients were classified into fast (n=29), intermediate (n=18) and slow responders (n=10) if they were smear-negative at 3, 5 or 8 weeks of treatment. alpha1-acid glycoprotein on enrolment and 1 week of treatment, alpha1-antitrypsin at week 1 and C-reactive protein and C3 after 3 weeks of therapy were higher in slow responders than in fast responders. alpha1-antitrypsin and alpha1-acid glycoprotein may be helpful in predicting treatment response at the time of initiation of therapy, and could be used as early markers to identify patients with an increased likelihood of treatment failure.

Host and Microbiological Factors Related to Dental Caries Development

Studies on dental caries suggest that in severe cases it may induce a systemic immune response. This occurs particularly when caries progresses into pulpal inflammation and results in abscess or fistula formation (AFF). We hypothesized that severe dental caries will affect the general health of children. The acute phase proteins alpha-1-acid glycoprotein (AGP), C-reactive protein (CRP) and the cytokine neopterin were chosen as parameters to monitor general health. Also, a polymorphism in the bacterial ligand CD14 (-260) was studied to investigate the relationship between genotype sensitivity for bacterial infections and AFF. In Suriname, children aged 6 years were recruited and enrolled into a dental care scheme, randomly assigned to 4 groups with different treatment strategies and monitored longitudinally. 348 children were included in the present study. Blood and saliva samples were taken at baseline and 1 year, and concentrations of serum AGP, CRP, neopterin, salivary Streptococcus mutans and CD14-260 C>T polymorphism were determined. There was no significant association between different treatment strategies and the serum parameters. Binary logistic regression analyses revealed a significant association between AFF as the outcome variable and the CD14 genotype and the concentrations of CRP and of neopterin as factors (p < 0.05). A significant negative association was found between the CD14-260 TT and AFF (p = 0.035, OR = 3.3) for the whole population. For children who had 4 or more carious lesions at baseline, the significance increased (p = 0.005, OR = 4.8), suggesting that the CD14-260 TT genotype was protective for AFF as a consequence of dental caries.

Isolation, characterization and quantitation of canine alpha-1-acid glycoprotein

The present study was designed to investigate the physicochemical properties of canine alpha-1-acid glycoprotein (AGP), and to establish a method for measuring serum AGP in canine. Fifty-three normal beagles were used in the study. AGP was purified from normal canine sera by successive ammonium sulfate precipitation, ion-exchange chromatography and gel filtration. The serum AGP concentration was measured by single radial immunodiffusion (SRID). Canine AGP had a molecular weight of 42,000 +/- 2,000 Da and contained 40.9% carbohydrate. Gel isoelectric focusing revealed microheterogeneity with 6-7 bands in a pI range (isoelectric points) of 3.2-3.8. AGP migrated to the alpha1-globulin region on immunoelectrophoresis. The serum AGP level of 35 normal beagles was 283.4 +/- 113.3 microg/ml. Canine AGP was isolated, and its physicochemical properties were clarified. SRID may be a useful method for quantification of serum AGP in canine.

Phase I and Pharmacokinetic Study of Imatinib Mesylate in Patients With Advanced Malignancies and Varying Degrees of Renal Dysfunction: A Study by the National Cancer Institute Organ Dysfunction Working Group

This study was undertaken to determine the safety, dose-limiting toxicities (DLT), maximum-tolerated dose (MTD), and pharmacokinetics of imatinib in cancer patients with renal impairment and to develop dosing guidelines for imatinib in such patients. Sixty adult patients with advanced solid tumors and varying renal function (normal, creatinine clearance [CrCL] >or= 60 mL/min; mild dysfunction, CrCL 40 to 59 mL/min; moderate dysfunction, CrCL 20 to 39 mL/min; and severe dysfunction, CrCL < 20 mL/min) received daily imatinib doses of 100 to 800 mg. Treatment cycles were 28 days long. The MTD was not reached for any group. DLTs occurred in two mild group patients (600 and 800 mg) and two moderate group patients (200 and 600 mg). Serious adverse events (SAEs) were more common in the renal dysfunction groups than in the normal group (P = .0096). There was no correlation between dose and SAEs in any group. No responses were observed. Several patients had prolonged stable disease. Imatinib exposure, expressed as dose-normalized imatinib area under the curve, was significantly greater in the mild and moderate groups than in the normal group. There was a positive correlation between serum alpha-1 acid glycoprotein (AGP) concentration and plasma imatinib, and an inverse correlation between plasma AGP concentration and imatinib clearance. Urinary excretion accounted for 3% to 5% of the daily imatinib dose. Daily imatinib doses up to 800 or 600 mg were well tolerated by patients with mild and moderate renal dysfunction, respectively, despite their having increased imatinib exposure.

Verification of Measurement of the Feline Serum Amyloid A (SAA) Concentration by Human SAA Turbidimetric Immunoassay and Its Clinical Application

Serum amyloid A (SAA) is one of the major acute phase proteins in cats that has potential to be used as an inflammatory marker. A previous study showed that the human SAA turbidimetric immunoassay (hSAA-TIA) could be used to measure the SAA concentration in cats. The objectives of the present study were to assess use of hSAA-TIA for determining the feline SAA concentration and to evaluate its clinical application. Recombinant feline SAA protein (rfSAA) was expressed in Escherichia coli and purified for SDS-PAGE and immunoblot analysis with anti-human SAA antibodies. The concentration of rfSAA was determined by ELISA and hSAA-TIA. Plasma SAA concentrations were measured in healthy and diseased cats by hSAA-TIA. The time-courses changes in the SAA and alpha1-acid glycoprotein (AGP) concentrations in the cats after ovariohysterectomy were investigated. In SDS-PAGE, rfSAA was detected as a clear band that reacted with anti-human SAA antibodies. There was significant correlation between the SAA concentration measured by ELISA and hSAA-TIA. The SAA concentration of the diseased cats (n=263) was significantly increased (P<0.01; 0.0-88.9 mg/l, mean: 7.52 mg/l) compared with that in the healthy cats (n=26; 0.0-0.9 mg/l, mean: 0.14 mg/l). No correlation was observed between SAA and WBC in the diseased cats. The SAA concentration changed more rapidly and remarkably than the AGP concentration after ovariohysterectomy. The present study revealed that hSAA-TIA is useful for determination of the feline SAA concentration. Measurement of the SAA concentration, in addition to the WBC count, would be clinically valuable as a routine test to detect inflammation.

Simultaneous Population Pharmacokinetic Model for Lopinavir and Ritonavir in HIV-Infected Adults

Lopinavir is a protease inhibitor indicated for the treatment of HIV infection. It is coformulated with low doses of ritonavir in order to enhance its pharmacokinetic profile. After oral administration, plasma concentrations of lopinavir can vary widely between different HIV-infected patients. To develop and validate a population pharmacokinetic model for lopinavir and ritonavir administered simultaneously in a population of HIV-infected adults. The model sought was to incorporate patient characteristics influencing variability in the drug concentration and the interaction between the two compounds. HIV-infected adults on stable therapy with oral lopinavir/ritonavir in routine clinical practice for at least 4 weeks were included. A concentration-time profile was obtained for each patient, and blood samples were collected immediately before and 1, 2, 4, 6, 8, 10 and 12 hours after a morning lopinavir/ritonavir dose. Lopinavir and ritonavir concentrations in plasma were determined by high-performance liquid chromatography. First, a population pharmacokinetic model was developed for lopinavir and for ritonavir separately. The pharmacokinetic parameters, interindividual variability and residual error were estimated, and the influence of different patient characteristics on the pharmacokinetics of lopinavir and ritonavir was explored. Then, a simultaneous model estimating the pharmacokinetics of both drugs together and incorporating the influence of ritonavir exposure on oral clearance (CL/F) of lopinavir was developed. Population analysis was performed using nonlinear mixed-effects modelling (NONMEM version V software). The bias and precision of the final model were assessed through Monte Carlo simulations and data-splitting techniques. A total of 53 and 25 Caucasian patients were included in two datasets for model building and model validation, respectively. Lopinavir and ritonavir pharmacokinetics were described by one-compartment models with first-order absorption and elimination. The presence of advanced liver fibrosis decreased CL/F of ritonavir by nearly half. The volume of distribution after oral administration (Vd/F) and CL/F of lopinavir were reduced as alpha1-acid glycoprotein (AAG) concentrations increased. CL/F of lopinavir was inhibited by ritonavir concentrations following a maximum-effect model (maximum inhibition [Imax] = 1, concentration producing 50% of the I(max) [IC50] = 0.36 mg/L). The final model appropriately predicted plasma concentrations in the model-validation dataset with no systematic bias and adequate precision. A population model to simultaneously describe the pharmacokinetics of lopinavir and ritonavir was developed and validated in HIV-infected patients. Bayesian estimates of the individual parameters of ritonavir and lopinavir could be useful to predict lopinavir exposure based on the presence of advanced liver fibrosis and the AAG concentration in an individual manner, with the aim of maximizing the chances of treatment success.

Pharmacokinetics of R- and S-Carvedilol in Routinely Treated Japanese Patients with Heart Failure

The purpose of this study was to evaluate the pharmacokinetics of R- and S-carvedilol in routinely treated Japanese patients with heart failure. We measured peak and trough blood concentrations at steady state following repeated oral administration to 24 patients. The blood concentration of S-carvedilol with potent beta-blocking activity was lower than that of R-carvedilol. The mean oral clearance (CL/F) of R- and S-carvedilol was not altered by CYP2D6*10, UGT2B7*3, and the etiology of heart failure. In addition, the CL/F values of enantiomers were not correlated with age, creatinine clearance, and plasma concentrations of alpha1-acid glycoprotein and brain natriuretic peptide. On the other hand, the mean CL/F values of R- and S-carvedilol in patients with heart failure were 0.89 and 1.52 l/h/kg, respectively, considerably lower than those estimated previously in healthy subjects. These results suggested that the pharmacokinetics of R- and S-carvedilol was altered significantly by heart failure.

Relationship between serum sialic acid and sialylated glycoproteins in alcoholics

Total sialic acid (TSA) has been suggested as a marker for chronic alcohol abuse. It seems that the elevation of TSA during excessive alcohol consumption reflects the changes in sialylated glycoproteins in the sera. On the other hand, chronic ethanol consumption increases the desialylation rate of many serum glycoproteins. The aim of this study was to evaluate the relationship between the total and free form of sialic acid levels (FSA), and the concentration of sialylated glycoproteins in alcoholics. We determined the serum concentration of many glycoproteins (alpha1-antitrypsin, alpha1-acid glycoprotein, haptoglobin, ceruloplasmin, transferrin, complement C3 protein, fibrinogen and immunoglobulin G) in a sample of 100 alcoholics and 30 healthy controls. Total sialic acid was determined by the enzymatic method and its free form by using a modification of the thiobarbituric acid method. Among alcoholics, we found increased concentrations of alpha1-antitrypsin and alpha1-acid glycoprotein but decreased levels of transferrin. The concentrations of TSA and FSA were significantly higher in alcoholics than in healthy controls. The tested glycoproteins, except for transferrin and immunoglobulin G, positively correlated with TSA and FSA. The correlations with TSA were higher than that with FSA. Chronic alcohol abuse alters the concentrations of some sialylated glycoproteins in the sera. The alpha1-antitrypsin, alpha1-acid glycoprotein, and transferrin are the only affected glycoproteins. The serum level of total and free form of sialic acid in the sera of alcoholics depends on the concentration of the most sialylated glycoproteins.