Usefulness of Belimumab in Adult Patients With Systemic Lupus Erythematosus Evaluated Using Single Indexes: A Meta-Analysis and Systematic Review.

Background: Belimumab is the first antibody drug approved for systemic lupus erythematosus (SLE), and is a fully human monoclonal antibody that inhibits soluble B lymphocyte stimulator protein. In clinical trials, a composite index was used to assess efficacy of belimumab. However, clinical guidelines on SLE treatment currently use single efficacy indexes. Objective: The main objective of this study was to perform a meta-analysis to evaluate the efficacy of belimumab utilizing single indexes used in routine clinical practice, rather than the composite efficacy index used in clinical trials during the development phase. As a secondary endpoint, safety was also evaluated. Methods: Several databases were searched to identify reports published up to December 1, 2021 on randomized controlled trials examining the efficacy of belimumab in adult patients with SLE. From the clinical trial data, efficacy was evaluated using single indexes including the SLE Disease Activity Index (SLEDAI), British Isles Lupus Assessment Group Index, and Physician Global Assessment. Safety was also assessed. Data were synthesized and analyzed using Review Manager 5.4. This study protocol was registered in the UMIN Clinical Trials Registry (Registration number: UMIN000052846). Results: The search identified 12 reports that met the inclusion criteria. Five reports were included in efficacy evaluation and 9 in safety evaluation. The primary endpoint was SLEDAI. Significantly more belimumab-treated patients achieved a ≥4-point reduction in SLEDAI (relative risk 1.28; 95% confidence interval, 1.16-1.40; P < 0.00001) compared with placebo. Other efficacy endpoints were also improved significantly in the belimumab group. No difference in safety was found between belimumab and placebo. Conclusions: The present meta-analysis evaluating clinical trial data using various single indexes recommended by clinical guidelines for SLE verifies that addition of belimumab to standard of care is efficacious for moderate-to-severe SLE.

Positive correlation between organic anion transporter 1B function indicated by plasma concentration of coproporphyrin-I and blood concentration of cyclosporin A in real-world patients.

AIMS: Cyclosporin A (CyA) has potent inhibitory activity on organic anion transporting polypeptide 1B (OATP1B), causing drug-drug interactions with its substrate drugs. 3-carboxy-4-methyl-5-propyl-2-furanpropionate (CMPF), a uraemic toxin, has also been suggested to inhibit OATP1B activity. Recent study has identified coproporphyrin-I (CP-I) as a specific endogenous substrate for OATP1B, which is useful to indicate OATP1B activity. We investigated the relationship of CP-I with CyA and CMPF concentrations in patients taking CyA. METHODS: In total, 121 blood samples from 74 patients who took CyA and underwent routine therapeutic drug monitoring were divided into trough and peak samples. RESULTS: CyA and CP-I concentrations were significantly higher in peak samples than in trough samples. A positive correlation between CP-I and CyA concentrations was found in all samples and in trough and peak samples, while no correlation was observed between CP-I and CMPF concentrations. Multiple regression analysis identified CyA and C-reactive protein concentrations as independent factors affecting CP-I concentration, with blood CyA concentration having markedly greater contribution to plasma CP-I concentration. CONCLUSION: The present study suggests that CyA inhibits OATP1B activity in a concentration-dependent manner in clinical setting, and that dose adjustment of OATP1B substrate drugs coadministered with CyA according to plasma CMPF concentration may not be necessary.

Evaluation of effects of indoxyl sulfate and parathyroid hormone on CYP3A activity considering the influence of CYP3A5 gene polymorphisms.

AIMS: Indoxyl sulfate and parathyroid hormone (PTH), which accumulate in chronic kidney disease (CKD), have been reported to reduce cytochrome P450(CYP)3A activity. Homozygotes of the CYP3A5*3 allele have reduced CYP3A5 activity compared to carriers of at least one CYP3A5*1 allele. 4ß-Hydroxycholesterol (4ß-OHC) has been established as an endogenous substrate reflecting CYP3A activity. 4ß-OHC is produced through hydroxylation by CYP3A4 and CYP3A5 and by autoxidation of cholesterol, whereas 4α-hydroxycholesterol (4α-OHC) is produced solely by autoxidation of cholesterol. This study focused on CKD patients and evaluated the effects of plasma indoxyl sulfate and intact-PTH concentrations on plasma 4ß-OHC concentration, 4ß-OHC/total cholesterol ratio and 4ß-OHC-4α-OHC, with consideration of the influence of CYP3A5 polymorphism. METHODS: Sixty-three CKD patients were analysed and divided into CYP3A5 carrier group (n = 26) and non-carrier group (n = 37). RESULTS: Plasma indoxyl sulfate significantly correlated inversely with 4ß-OHC concentration and with 4ß-OHC-4α-OHC in both the CYP3A5*1 carrier group (r = -0.42, P = .034; r = -0.39, P = .050, respectively) and the non-carrier group (r = -0.45, P = .0054; r = -0.39, P = .019, respectively). However, multiple regression analysis did not identify plasma indoxyl sulfate concentration as a significant independent factor associated with any of the CYP3A activity indices. There was no significant correlation between plasma intact-PTH concentration and any of the CYP3A activity indices. CONCLUSIONS: The present results suggest that plasma indoxyl sulfate and intact-PTH concentrations do not have clinically significant effects on CYP3A activity in patients with CKD.

Sensitive UHPLC-MS/MS quantification method for 4ß- and 4α-hydroxycholesterol in plasma for accurate CYP3A phenotyping.

4ß-Hydroxycholesterol (4ß-OHC) is formed by Cytochrome P450 (CYP)3A and has drawn attention as an endogenous phenotyping probe for CYP3A activity. However, 4ß-OHC is also increased by cholesterol autooxidation occurring in vitro due to dysregulated storage and in vivo by oxidative stress or inflammation, independent of CYP3A activity. 4α-hydroxycholesterol (4α-OHC), a stereoisomer of 4ß-OHC, is also formed via autooxidation of cholesterol, not by CYP3A, and thus may have clinical potential in reflecting the state of cholesterol autooxidation. In this study, we establish a sensitive method for simultaneous quantification of 4ß-OHC and 4α-OHC in human plasma using ultra-high performance liquid chromatography coupled to tandem mass spectrometry. Plasma samples were prepared by saponification, two-step liquid-liquid extraction, and derivatization using picolinic acid. Intense [M+H]+ signals for 4ß-OHC and 4α-OHC di-picolinyl esters were monitored using electrospray ionization. The assay fulfilled the requirements of the US Food and Drug Administration guidance for bioanalytical method validation, with a lower limit of quantification of 0.5 ng/ml for both 4ß-OHC and 4α-OHC. Apparent recovery rates from human plasma ranged from 88.2% to 101.5% for 4ß-OHC, and 91.8% to 114.9% for 4α-OHC. Additionally, matrix effects varied between 86.2% and 117.6% for 4ß-OHC and between 89.5% and 116.9% for 4α-OHC. Plasma 4ß-OHC and 4α-OHC concentrations in healthy volunteers, stage 3-5 chronic kidney disease (CKD) patients, and stage 5D CKD patients as measured by the validated assay were within the calibration ranges in all samples. We propose this novel quantification method may contribute to accurate evaluation of in vivo CYP3A activity.

Perianesthetic mortality in English Bulldogs: a retrospective analysis in 2010 - 2017.

BACKGROUND: Many veterinarians consider English Bulldogs to have a greater perianesthetic mortality risk. The aims of this study were to 1) determine total and anesthesia-related, perianesthetic mortality (PAM) rates in English Bulldogs (EB), 2) identify potential risk factors associated with mortality in EB, and 3) determine the difference in the perianesthetic mortality rates between EB, other-brachycephalic breeds (OB), and non-brachycephalic breeds (NB). Records from EB that were anesthetized between 2010 and 2017, were investigated. OB and NB were enrolled to match with each EB based on a procedure and age from the study period. Data collected in EB included: age, ASA status, weight, procedure types, anesthetic and analgesic management, anesthetic duration, anesthetic recovery location, and cause of death. Age and cause of death were determined from OB and NB. Fisher's exact test was used to compare PAM rate and age in EB, OB, and NB. Mann-Whitney U test was used to compare EB survivor and EB non-survivor. Logistic regression models were used to identify factors and odds ratio (OR) associated with PAM in EB. RESULT: Two hundred twenty nine EB, 218 OB, and 229 NB were identified. The total and anesthesia-related PAM rates in EB were 6.6 and 3.9%, respectively. EB had a greater total PAM rate compared with OB (p = 0.007). ASA status was different between survivors and non-survivors in EB (p < 0.01). Risk factors identified regardless of the cause of death were premedication with full µ opioids (OR = 0.333, p = 0.114), continuous infusion of ketamine post-operatively (OR = 13.775, p = 0.013), and acepromazine administration post-operatively (OR = 7.274, p = 0.004). The most common cause of death in EB was postoperative respiratory dysfunction (87.5%). CONCLUSION: Total and anesthesia-related mortality in EB is considerable. Most deaths in EB occurred during the postoperative period secondary to respiratory complications.

Highly sensitive simultaneous quantification of indoxyl sulfate and 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid in human plasma using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry.

Indoxyl sulfate and 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid are uremic toxins that accumulate in renal failure and have been reported to decrease the activities of the drug-metabolizing enzyme cytochrome P450 3A and the drug transporter organic anion transporting polypeptides 1B, respectively. In this study, we established and validated an assay for simultaneous quantification of indoxyl sulfate and 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid in human plasma. The samples were pretreated by solid-phase extraction, and measured by ultra-high-performance liquid chromatography-tandem mass spectrometry. The validation results for this assay were within the acceptable limits recommended by the US Food and Drug Administration, with a lower limit of quantitation of 0.05 µg/mL for both indoxyl sulfate and 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid. Recovery rates of indoxyl sulfate and 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid corrected by internal standard were 100.7-101.9 and 100.2-101.3%, respectively. Matrix effects of indoxyl sulfate and 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid corrected by internal standard were 101.1-105.5 and 97.0-103.8%, respectively. The validated assay was used to analyze indoxyl sulfate and 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid concentrations in the plasma samples of healthy volunteers and patients with chronic kidney disease. All the measured plasma indoxyl sulfate and 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid concentrations were within the calibration ranges. This novel method may contribute to predicting the activities of drug-metabolizing enzymes and drug transporters in individual patients.

Pharmacokinetics and pharmacodynamic effects in koi carp (Cyprinus carpio) following immersion in propofol.

OBJECTIVE: To test the hypothesis that plasma propofol concentration (PPC) is associated with anesthetic effect in koi carp administered propofol by immersion. STUDY DESIGN: Prospective study. ANIMALS: Twenty mature koi carp (mean ± standard deviation, 409.4 ± 83.7 g). METHODS: Fish were immersed in propofol (5 mg L-1). Physiological variables and induction and recovery times were recorded. In phase I, blood was sampled for PPC immediately following induction and at recovery. In phase II, following induction, fish were maintained with propofol (4 mg L-1) via a recirculating system for 20 minutes. Following established induction, blood was sampled at 1, 10 and 20 minutes. In phase III (n = 19), fish were anesthetized as in phase II with blood sampled nine times in a sparse sampling strategy. Simultaneously, a pharmacodynamics rubric was used to evaluate anesthetic depth. PPC was determined using high performance liquid chromatography with fluorescence detection. Following evaluation of normality, data were analyzed using paired t test or Spearman correlation test (significance was set at p < 0.05). RESULTS: In phase I, mean PPCs at induction (20.12 µg mL-1) and recovery (11.62 µg mL-1) were different (p < 0.001). In phase II, only mean PPCs at induction (17.92 µg mL-1) and 10 minutes (21.50 µg mL-1) were different (p = 0.013). In phase III, a correlation between PPCs and the pharmacodynamic rubric scores was found (p < 0.001, r = -0.93). There was no correlation between PPCs and recovery time (p = 0.057, r = 0.433). A two-compartment open model was chosen for the pharmacokinetic model. Absorption rate constant, elimination rate constant and intercompartmental rate constant were 0.48, 0.006 and 0.02 minute-1, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Measurable PPCs were achieved in koi carp anesthetized with propofol by immersion. Anesthetic depth of fish was negatively correlated with PPCs, but recovery time was not.

Electrical resistivity tomography combined with seismic data estimates heterogeneous distribution of near-seafloor concentrated gas hydrates within gas chimneys.

Near-seafloor concentrated gas hydrates (GHs) containing large amounts of methane have been identified at various gas chimney sites. Although understanding the spatial distribution of GHs is fundamental for assessing their dissociation impact on aggravating global warming and resource potential, the spatial distribution of GHs within gas chimneys remains unclear. Here, we estimate the subseafloor distribution of GHs at a gas chimney site in the Japan Sea using marine electrical resistivity tomography data. The resulting two-dimensional subseafloor resistivity structure shows high anomalies (10-100 Ωm) within seismically inferred gas chimneys. As the resistivity anomalies are aligned with high amplitude seismic reflections and core positions recovering GHs, we interpret the resistivity anomalies are near-seafloor concentrated GH deposits. We also detect various distribution patterns of the high resistivity anomalies including 100-m wide and 40-m thick anomaly near the seafloor and 500-m wide anomaly buried 50 m below the seafloor, suggesting that GHs are heterogeneously distributed. Therefore, considering such heterogeneous GH distribution within gas chimneys is critical for in-depth assessments of GH environmental impacts and energy resources.

Evaluation of intra- and inter-individual variations in plasma belimumab concentrations in adult patients with systemic lupus erythematosus.

In this study, plasma belimumab concentrations were measured over the course of treatment in systemic lupus erythematosus (SLE) patients on belimumab therapy, and intra- and interindividual variations in plasma belimumab concentration were evaluated. A single-center prospective study was conducted at Oita University Hospital to evaluate trough plasma concentrations over the course of treatment in 13 SLE patients treated with intravenous belimumab. Plasma belimumab concentrations were measured by a validated ultra-high performance liquid chromatography with tandem mass spectrometry method. The median age of the patients was 40 (interquartile range: 35-51) years and the median weight was 51.8 (47.0-58.1) kg. A mean of 9.4 (range: 1-13) blood samples was collected per patient at routine visits. The mean (± SD) plasma belimumab concentration was 33.4 ± 11.9 µg/mL in the patient with the lowest concentration and 170.0 ± 16.6 µg/mL in the patient with the highest concentration, indicating a 5-fold difference between patients. On the other hand, the within-patient coefficient of variation ranged from 7.1% to 35.7%, showing no large variations. No significant correlation was observed between plasma belimumab concentration and belimumab dose (mg/kg) (Spearman's rank correlation coefficient = 0.22, p = .54). Examinations of trough plasma belimumab concentrations over the course of treatment in patients with SLE showed small intraindividual variation but large interindividual variation. Plasma belimumab trough concentration varied widely among patients administered the approved dose.

Relationship of plasma 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid concentration with OATP1B activity in patients with chronic kidney disease.

Organic anion-transporting polypeptides (OATP)1B are drug transporters mainly expressed in the sinusoidal membrane. Many studies have suggested that OATP1B activity is affected by genetic factor, the uremic toxin 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF), and inflammatory cytokines, such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Coproporphyrin-I (CP-I) is spotlighted as a highly accurate endogenous substrate of OATP1B. We previously reported a positive correlation between plasma CMPF and CP-I concentrations in patients with chronic kidney disease (CKD). The present study evaluated the impact of genetic polymorphisms, CMPF, IL-6, TNF-α, and estimated glomerular filtration rate (eGFR) on individual differences in OATP1B activity in patients with CKD. Seventy-three patients with CKD who received kidney transplant at least 3 months earlier were analyzed. Plasma CP-I concentration was higher in OATP1B1*15 carriers than in non-carriers. In all patients, CP-I did not correlate significantly with CMPF, IL-6, TNF-α, or eGFR. However, when the dataset was cut off at CMPF concentration of 8 and 7 µg/mL, 4 µg/mL, 3 µg/mL or 2 µg/mL, CMPF correlated positively with CP-I, and correlation coefficient tended to be higher as plasma CMPF concentration was lower. In conclusion, OATP1B1*15 impacted OATP1B activity in patients with CKD, but IL-6 and TNF-α did not. However, the impact of CMPF on OATP1B activity was limited to low CMPF concentrations, and the effect could be saturated at high concentrations. When prescribing an OATP1B substrate drug for patients with CKD, the OATP1B1*15 carrier status and plasma CMPF concentration may need to be considered to decide the dose regimen.

Rapid and simple quantification of belimumab in human plasma using ultra-high performance liquid chromatography with tandem mass spectrometry.

OBJECTIVE: Belimumab is a monoclonal antibody against the B-lymphocyte stimulating factor and is approved for the treatment of patients with systemic lupus erythematosus (SLE) not responding adequately to existing therapies. In this study, we established and validated an assay for quantifying belimumab in human plasma. METHODS: From the peptides generated by trypsin digestion of belimumab, in silico analysis was used to search for unique peptides to determine the surrogate peptides. Samples were trypsin digested, pretreated with solid phase extraction, and analyzed by ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) to quantify the surrogate peptide in the samples. The assay was validated according to the Food and Drug Administration (FDA) bioanalytical method validation guidance. We used the established assay to quantify plasma belimumab concentrations in two SLE patients treated with belimumab. RESULTS: Among the unique peptides identified by the in silico analysis, the peptide with the best peak shape when measured by UHPLC-MS/MS was selected as the surrogate peptide. The validation results of this assay met the acceptable criteria recommended by the FDA guidance. The lower limit of quantification (LLOQ) for belimumab was 2 µg/mL. Recovery rates and matrix effects when corrected for internal standards were 91.5-114.3 % and 96.9-108.4 %, respectively. Plasma concentrations of belimumab were measured in 12 samples from two belimumab-treated SLE patients. All concentrations were within the calibration range. CONCLUSIONS: We have established and validated a method for measuring plasma belimumab concentrations using UHPLC/MS-MS. By measuring plasma belimumab concentrations in more patients, this method is expected to contribute to appropriate use of belimumab.

Evaluation of the usefulness of plasma 4ß-hydroxycholesterol concentration normalized by 4α-hydroxycholesterol for accurate CYP3A phenotyping.

Plasma 4ß-hydroxycholesterol (OHC) has drawn attention as an endogenous substrate indicating CYP3A activity. Plasma 4ß-OHC is produced by hydroxylation by CYP3A4 and CYP3A5 and by cholesterol autoxidation. Plasma 4α-OHC is produced by cholesterol autoxidation and not affected by CYP3A activity. This study aimed to evaluate the usefulness of plasma 4ß-OHC concentration minus plasma 4α-OHC concentration (4ß-OHC-4α-OHC) compared with plasma 4ß-OHC concentration and 4ß-OHC/total cholesterol (TC) ratio in cross-sectional evaluation of CYP3A activity. Four hundred sixteen general adults were divided into 191 CYP3A5*1 carriers and 225 non-carriers. Twenty-six patients with chronic kidney disease (CKD) with CYP3A5*1 allele were divided into 14 with CKD stage 3 and 12 with stage 4-5D. Area under the receiver operating characteristic curve (AUC) for the three indices were evaluated for predicting presence or absence of CYP3A5*1 allele in general adults, and for predicting CKD stage 3 or stage 4-5D in patients with CKD. There was no significant difference between AUC of 4ß-OHC-4α-OHC and AUC of plasma 4ß-OHC concentration in general adults and in patients with CKD. AUC of 4ß-OHC-4α-OHC was significantly smaller than that of 4ß-OHC/TC ratio in general adults (p = 0.025), but the two indices did not differ in patients with CKD. In conclusion, in the present cross-sectional evaluation of CYP3A activity in general adults and in patients with CKD with CYP3A5*1 allele, the usefulness of 4ß-OHC-4α-OHC was not different from plasma 4ß-OHC concentration or 4ß-OHC/TC ratio. However, because of the limitations in study design and subject selection of this research, these findings require verification in further studies.

Soymorphin-5, a soy-derived µ-opioid peptide, decreases glucose and triglyceride levels through activating adiponectin and PPARα systems in diabetic KKAy mice.

Soymorphin-5 (YPFVV) derived from soybean ß-conglycinin ß-subunit is a µ-opioid agonist peptide having anxiolytic-like activity. Here, we show that soymorphin-5 improves glucose and lipid metabolism after long-term oral administration to KKAy mice, a type 2 diabetes model animal. Soymorphin-5 inhibited hyperglycemia without an increase in plasma insulin levels in KKAy mice. Soymorphin-5 also decreased plasma and liver triglyceride (TG) levels and liver weight, suggesting that soymorphin-5 improved lipid metabolism. Soymorphin-5 increased plasma adiponectin concentration and liver mRNA expression of AdipoR2, a subtype of adiponectin receptor that is involved in stimulating the peroxisome proliferator-activated receptor (PPAR)α pathway and fatty acid ß-oxidation. The expressions of the mRNA of PPARα and its target genes acyl-CoA oxidase, carnitine palmitoyltransferase 1 A, and uncoupling protein-2, in the liver were also increased after oral administration of soymorphin-5. Furthermore, des-Tyr-soymorphin-5 (PFVV) without µ-opioid and anxiolytic-like activities did not decrease blood glucose levels in KKAy mice. These results suggest that µ-opioid peptide soymorphin-5 improves glucose and lipid metabolism via activation of the adiponectin and PPARα system and subsequent increases of ß-oxidation and energy expenditure in KKAy mice.

Characterization of ovolin, an orally active tryptic peptide released from ovalbumin with anxiolytic-like activity.

We found that tryptic digest of ovalbumin after oral (p.o.) and intraperitoneal (i.p.) administration exhibited anxiolytic-like activity in mice, and then searched for orally active low-molecular-weight peptides with anxiolytic-like activity in the tryptic digest. Val-Tyr-Leu-Pro-Arg, named ovolin, corresponding to ovalbumin (280-284), mimicked the anxiolytic-like activity after p.o. and i.p. administration. The anxiolytic-like activity of ovolin was inhibited by indomethacin, a cyclooxygenase (COX) inhibitor, or BWA868C, an antagonist of the DP1 receptor for prostaglandin (PG) D2 . Ovolin-induced anxiolytic-like activity was also blocked by SCH58261 or bicuculline, antagonists of the adenosine A2A and GABAA receptors, respectively. Ovolin has no affinity for the DP1 , A2A and GABAA receptors. Taken together, ovolin may exhibit anxiolytic-like activity in a manner dependent on the PGD2 -DP1 system coupled to the A2A and GABAA receptors.

ß-Lactotensin derived from bovine ß-lactoglobulin exhibits anxiolytic-like activity as an agonist for neurotensin NTS(2) receptor via activation of dopamine D(1) receptor in mice.

ß-Lactotensin (His-Ile-Arg-Leu) is a bioactive peptide derived from bovine milk ß-lactoglobulin, acting as a natural agonist for neurotensin receptors. We found that ß-lactotensin exhibited anxiolytic-like activity in an elevated plus-maze test after its intraperitoneal (i.p.) administration in mice. ß-Lactotensin was also orally active. The anxiolytic-like activity of ß-lactotensin after i.p. administration was blocked by levocabastine, an antagonist for the neurotensin NTS(2) receptor. ß-Lactotensin had anxiolytic-like activity in wild-type but not Ntsr2-knockout mice. ß-Lactotensin increased intracellular Ca(2+) flux in glial cells derived from wild-type mice but not Ntsr2 knockout mice. These results suggest that ß-lactotensin acts as an NTS(2) receptor agonist having anxiolytic-like activity. The anxiolytic-like activity of ß-lactotensin was also blocked by SCH23390 and SKF83566, antagonists for dopamine D(1) receptor, but not by raclopride, an antagonist for D(2) receptor. Taken together, ß-lactotensin may exhibit anxiolytic-like activity via NTS(2) receptor followed by D(1) receptor.

Factors Influencing Plasma Coproporphyrin-I Concentration as Biomarker of OATP1B Activity in Patients With Rheumatoid Arthritis.

Organic anion transporting polypeptides (OATPs) 1B are drug transporters mainly expressed in the sinusoidal membrane. In previous reports, genetic factor, 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF), which is one of the uremic toxins, inflammatory cytokines such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) decreased OATP1B1 activity in vitro, but in vivo effects of these factors have not been elucidated. Plasma coproporphyrin-I (CP-I) is spotlighted as a highly accurate endogenous substrate of OATP1B. This study focused on patients with rheumatoid arthritis (RA) and evaluated the influence of several factors comprising gene polymorphisms, uremic toxins, and inflammatory cytokines on OATP1B activity using plasma CP-I concentration. Thirty-seven outpatients with RA who satisfied the selection criteria were analyzed at the time of recruitment (baseline) and at the next visit. OATP1B1*15 carriers tended to have higher CP-I concentration compared with noncarriers. Plasma CP-I correlated positively with CMPF concentration, but did not correlate with IL-6 or TNF-α concentration. Multiple logistic regression analysis by stepwise selection identified plasma CMPF concentration and OATP1B1*15 allele as significant factors independently affecting plasma CP-I concentration at baseline and at the next visit, respectively. In conclusion, the present results suggest that inflammatory cytokines do not have clinically significant effects on OATP1B activity, whereas the effects of genetic polymorphisms and uremic toxins should be considered.

Bilateral cataract surgery in a Japanese macaque (Macaca fuscata): A case report.

A 7.5-year-old intact male Japanese macaque was presented for evaluation of vision loss. After a complete ophthalmic examination, the patient was diagnosed with hypermature cataract in both eyes. After the cataract surgery, it was able to locate food and walk in a straight line.

Relationship of hemoglobin level and plasma coproporphyrin-I concentrations as an endogenous probe for phenotyping OATP1B.

Plasma coproporphyrin-I (CP-I) concentration is used as a sensitive and selective endogenous probe for phenotyping organic anion transporting polypeptides 1B (OATP1B) activity in many studies. CP-I is produced in the process of heme synthesis, but the relationship between plasma CP-I concentrations and heme synthesis activity is unknown. In this study, we evaluated the relationship between plasma CP-I concentration and hemoglobin level as a biomarker of heme synthesis activity. The data of 391 subjects selected from the Japanese general population were analyzed. One hundred twenty-six participants had OATP1B1*15 allele, 11 of whom were homozygous (OATP1B1*15/*15). Multiple regression analysis identified hemoglobin level as an independent variable associated with plasma CP-I concentration (p < 0.0001). A significant positive correlation was observed between hemoglobin level and plasma CP-I concentration in participants without OATP1B1*15 allele (n = 265; rs  = 0.35, p < 0.0001) and with OATP1B1*15 allele (n = 126; rs  =0.27, p = 0.0022). However, Kruskal-Wallis test showed no large difference in Kruskal-Wallis statistics between the distribution of plasma CP-I concentrations and that of ratio of plasma CP-I to hemoglobin among six OATP1B1 polymorphism groups. These findings suggest that the hemoglobin level seems to reflect biosynthesis of CP-I. However, correction by hemoglobin level is not required when using basal plasma CP-I concentration for phenotyping OATP1B activity.

Use of tricaine methanesulfonate or propofol for immersion euthanasia of goldfish (Carassius auratus).

OBJECTIVE To substantiate current AVMA guidelines for immersion euthanasia of goldfish (Carassius auratus) with tricaine methanesulfonate (TMS), determine whether immersion in propofol at 5 times its immersion anesthesia concentration for 30 minutes is sufficient for euthanasia of goldfish, and quantify the duration of myocardial contraction following immersion of goldfish in TMS and decapitation. DESIGN Prospective clinical trial. ANIMALS 36 healthy, adult goldfish. PROCEDURES Goldfish were randomly assigned to be immersed in 1 of 6 test solution treatments (n = 6/treatment): TMS (500 mg/L) for 15 minutes followed by placement in anesthetic agent-free water (T15W), placement out of water (T15A), or decapitation (T15D); TMS (1,000 mg/L) for 15 minutes followed by placement in anesthetic agent-free water (T15XW); TMS (500 mg/L) for 30 minutes followed by placement in anesthetic agent-free water (T30W); or propofol (25 mg/L) for 30 minutes followed by placement in anesthetic agent-free water (P30W). Any fish that resumed operculation in group T15A was returned to anesthetic agent-free water. Times from onset of immersion to induction of anesthesia, cessation and resumption of operculation, and recovery (T15W, T15A, T15XW, T30W, P30W) or cessation of Doppler ultrasounds (T15D) were recorded. RESULTS Overall, 5 of 6, 6 of 6, 6 of 6, 6 of 6, and 5 of 6 fish survived in the T15W, T15A, T15XW, T30W, and P30W groups, respectively. Median time to cessation of Doppler ultrasounds in group T15D was 77.5 minutes (range, 30 to 240 minutes). CONCLUSIONS AND CLINICAL RELEVANCE Timed immersion in test solutions (TMS at 500 mg/L or 1,000 mg/L or propofol at 25 mg/L) resulted in death in only 7% (2/30) of immersed goldfish. Myocardial contractions continued for up to 4 hours in decapitated goldfish.

Physiologic and biochemical assessments of koi (Cyprinus carpio) following immersion in propofol.

OBJECTIVE: To determine efficacy of propofol as an immersion agent to induce general anesthesia in koi (Cyprinus carpio). DESIGN: Prospective, crossover study. ANIMALS: 10 adult koi (mean ± SD weight, 325 ± 81 g). PROCEDURES: Koi were exposed to each of 4 concentrations of propofol (1, 2.5, 5, and 10 mg/L) with a 1-week washout period between trials. In a subsequent trial, koi were anesthetized with propofol (5 mg/L) and anesthesia was maintained with propofol (3 mg/L) for 20 minutes. Response to a noxious stimulus was assessed by means of needle insertion into an epaxial muscle. RESULTS: At a propofol concentration of 1 mg/L, koi were sedated but never anesthetized. At propofol concentrations of 2.5, 5, and 10 mg/L, mean ± SD anesthetic induction times were 13.4 ± 3.3, 3.8 ± 1.1, and 2.3 ± 0.9 minutes, respectively; mean recovery times were 12.9 ± 8.3, 11.0 ± 6.3, and 18.1 ± 13.0 minutes; mean heart rates were 57 ± 25, 30 ± 14, and 22 ± 14 beats/min; mean opercular rates were 58 ± 18, 68 ± 15, and 48 ± 22 beats/min; and 1 of 10, 2 of 10, and 0 of 10 fish responded to needle insertion. All fish recovered satisfactorily. Following 20 minutes of anesthesia, 2 fish had recovery times > 4 hours and 1 fish died. CONCLUSIONS AND CLINICAL RELEVANCE: Immersion in propofol at concentrations ≥ 2.5 mg/L induced general anesthesia in koi. Maintenance of anesthesia with propofol for 20 minutes was associated with prolonged recovery times in 2 of 9 and death in 1 of 9 koi.