Comparison of electrochemiluminescence immunoassay and latex agglutination turbidimetric immunoassay for evaluation of everolimus blood concentrations in renal transplant patients.

WHAT IS KNOWN AND OBJECTIVE: For analysis of blood concentrations of everolimus, many hospital laboratories use either latex agglutination turbidimetric immunoassay (LTIA) or electrochemiluminescence immunoassay (ECLIA). However, no studies have compared both immunoassay methods under the same conditions. Accordingly, in this study, we compared everolimus blood concentrations obtained by LTIA and ECLIA in renal transplant patients. METHODS: Blood samples (n = 230) from 60 renal transplant patients (19 female and 41 male) were evaluated using both immunoassays. Subsequently, we switched the assay for detection of everolimus blood concentrations from LTIA to ECLIA as a clinical application. Three quality control (QC) samples for LTIA were analysed using ECLIA, and 3 QC samples for ECLIA were analysed using LTIA. RESULTS: The Deming regression of ECLIA versus LTIA generated the following parameters: slope, 1.0067 and intercept, 1.7489 ng/mL, in the analysis of 230 samples. Bland-Altman analysis showed an average positive bias of 1.73 ng/mL between ECLIA and LTIA. When the clinical apparatus was switched from LTIA to ECLIA, the average everolimus blood concentration assayed by LTIA before switching was 3.57 ng/mL, whereas that by ECLIA after switching in the same patients taking the same daily dose (mean: 1.43 mg/day) was 5.85 ng/mL. The QCs assayed using LTIA were lower by an average of 67.3% (range: 55.8%-79.5%) for ECLIA, and in the same 230 samples from patients, the everolimus blood concentrations assayed by LTIA were lower by an average of 67.4% (range: 37.1%-114.5%) of ECLIA. WHAT IS NEW AND CONCLUSION: Analysis of everolimus concentrations by immunoassays with high precision and accuracy is required to ensure long-term survival of transplant recipients. Although the concentrations of QCs and calibrators of everolimus in LTIA were previously corrected to 70% concentration because of cross-reactivity with everolimus metabolites, these adjustments may need to be reviewed.

Multiple inductive effects of carbamazepine on combined therapy with paliperidone and amlodipine.

WHAT IS KNOWN AND OBJECTIVE: Carbamazepine is a potent inducer of cytochrome P450 3A and P-glycoprotein. However, there are no reports of the effects of carbamazepine on more than one co-administered drug. CASE SUMMARY: A 53-year-old female patient with schizophrenia and hypertension was on paliperidone 12 mg/day and amlodipine 5 mg/day. When carbamazepine was added to this prescription, the plasma concentrations of both drugs decreased dramatically in a dose-dependent manner. Although the patient's psychotic symptoms did not change, as a result, her mean blood pressure increased to 160·1/103·6 mmHg from 138·4/91·4 mmHg at a carbamazepine dose of 600 mg/day. WHAT IS NEW AND CONCLUSION: Theses cases show the effect of carbamazepine induction on two drugs simultaneously. Care is required when carbamazepine is added to drug regimens including paliperidone or amlodipine alone or together.

Effects of multiple-dose rifampicin 450 mg on the pharmacokinetics of fexofenadine enantiomers in Japanese volunteers.

WHAT IS KNOWN AND OBJECTIVE: Rifampicin is a potent inducer of P-glycoprotein (P-gp) and inhibitor of organic anion-transporting polypeptides (OATPs), with fexofenadine acting as a substrate for both mechanisms. Simultaneous administration of single- or multiple-dose rifampicin 600 mg significantly increases the concentrations of fexofenadine enantiomers by inhibiting OATP transporters. However, the effects of rifampicin 450 mg are unknown. Here, we evaluated the effects of multiple doses of rifampicin 450 mg on the pharmacokinetics of fexofenadine enantiomers in healthy Japanese volunteers. METHODS: In this randomized, two-phase, double-blind crossover study, 10 healthy volunteers received rifampicin 450 mg/day or placebo for 7 days. On day 7, fexofenadine 60 mg was co-administered simultaneously. RESULTS AND DISCUSSION: Rifampicin significantly increased the mean area under the plasma concentration-time curve (AUC) of (R)- and (S)-fexofenadine (3.10-fold and 3.48-fold, respectively) and decreased the renal clearance of (R)- and (S)-fexofenadine (0.40-fold and 0.47-fold, respectively), causing marked differences in the mean amounts of these enantiomers excreted into the urine in the rifampicin phase (P < 0.001). These results indicated that multiple doses of rifampicin 450 mg may be sufficient to inhibit the renal influx transporter and OATP-mediated hepatic uptake of both enantiomers. Moreover, these effects may be greater than the P-gp-inductive effects of rifampicin. Therefore, the interactive mechanism of multidose rifampicin may occur through a combination of OATP and P-gp transporters, thereby altering the pharmacokinetics of fexofenadine enantiomers. WHAT IS NEW AND CONCLUSIONS: In this study of rifampicin 450 mg, the interactive magnitude of the mean AUC values of fexofenadine enantiomers was higher than that observed in the previous study of rifampicin 600 mg, and no dose-dependent inhibitory effects of rifampicin were observed. These effects may be clinically significant in patients receiving fexofenadine and rifampicin.

Cyclic compressive loading on 3D tissue of human synovial fibroblasts upregulates prostaglandin E2 via COX-2 production without IL-1ß and TNF-α.

OBJECTIVE: Excessive mechanical stress on synovial joints causes osteoarthritis (OA) and results in the production of prostaglandin E2 (PGE2), a key molecule in arthritis, by synovial fibroblasts. However, the relationship between arthritis-related molecules and mechanical stress is still unclear. The purpose of this study was to examine the synovial fibroblast response to cyclic mechanical stress using an in vitro osteoarthritis model. METHOD: Human synovial fibroblasts were cultured on collagen scaffolds to produce three-dimensional constructs. A cyclic compressive loading of 40 kPa at 0.5 Hz was applied to the constructs, with or without the administration of a cyclooxygenase-2 (COX-2) selective inhibitor or dexamethasone, and then the concentrations of PGE2, interleukin-1ß (IL-1ß), tumour necrosis factor-α (TNF-α), IL-6, IL-8 and COX-2 were measured. RESULTS: The concentrations of PGE2, IL-6 and IL-8 in the loaded samples were significantly higher than those of unloaded samples; however, the concentrations of IL-1ß and TNF-α were the same as the unloaded samples. After the administration of a COX-2 selective inhibitor, the increased concentration of PGE2 by cyclic compressive loading was impeded, but the concentrations of IL-6 and IL-8 remained high. With dexamethasone, upregulation of PGE2, IL-6 and IL-8 was suppressed. CONCLUSION: These results could be useful in revealing the molecular mechanism of mechanical stress in vivo for a better understanding of the pathology and therapy of OA. Cite this article: Bone Joint Res 2014;3:280-8.

Prolonged matrix metalloproteinase-3 high expression after cyclic compressive load on human synovial cells in three-dimensional cultured tissue.

Excessive mechanical stress is thought to be a factor in the development of joint disorders through the expression of matrix metalloproteinases (MMPs) and related cytokines. Although studies revealed that mechanical stress on the synovium induces MMP expression, it is still not known which MMPs prolonged high level expression. The authors focused on MMP-3, which is one of the major factors in joint disorders such as rheumatism and temporomandibular joint disorders. They examined mRNA and protein levels of MMP-3, other MMPs and related cytokines after loading stress. Human synovial cells were seeded onto a collagen scaffold and different magnitudes of cyclic compressive load were applied for 1h. Time-dependent mRNA and protein levels for catabolic genes were examined after loading. mRNA expressions of MMP-1, MMP-3, MMP-9, IL-6, IL-8 and IL-1ß increased after excessive compression. In particular, only mRNA of MMP-3 was up-regulated and maintained at a high level for 24h after excessive loading. The concentrations of MMP-3, IL-6 and IL-8 in culture media after loading increased with excessive compression. These results may account for the pathomechanism of MMP-3 induced by cyclic load on synovial cells in joint disorders.

Treatment effects of Fast ForWord demonstrated by magnetoencephalography (MEG) in a child with developmental dyslexia.

Treatment effects of Fast ForWord, hypothesized to ameliorate temporal processing deficits, were demonstrated by magnetoencephalography in a child with dyslexia using four paradigms: Word/Non-word Reading (NW), Grapheme-to-Phoneme Matching (GP), Verbal, and Spatial Working Memory (VWM, SWM). Shifts in brain activation from right inferior frontal and temporal to left frontal, bilateral supramarginal, and transverse temporal regions occurred during GP. During NW, shifts progressed from (1) right or bilateral anterior and superior to (2) left, inferior frontal, to (3) left, superior posterior temporoparietal, to (4) left, inferior, posterior temporooccipital regions. Reading and written language improvements were noted in passage comprehension and spelling.

Rate of eating and body weight in patients with type 2 diabetes or hyperlipidaemia.

This preliminary investigation, involving 422 patients, tested the hypothesis that rate of eating is associated with obesity in patients with type 2 diabetes or hyperlipidaemia at all ages. The patients' eating habits were determined using a questionnaire, and the patients were classified as quick, normal or slow eaters. The body mass indices of the three groups were compared. The body mass indices of the male patients who ate quickly (25.4 +/- 0.2 kg/m2) were significantly higher than those of the patients who ate at a normal rate (24.4 +/- 0.3 kg/m2) or slowly (24.1 +/- 0.5 kg/m2). No difference between body mass indices in the female groups was found. It was speculated that rate of eating affects body weight in male patients with type 2 diabetes or hyperlipidaemia.

[Two cases of liver scar that was examined by biliary scintigraphy].

36-year-old-female admitted because of jaudice and ascites. T-bil was 18.5 mg/dl and transaminase, ALP, LDH and gamma-GTP was elevated. Ultrasonography (US) showed that right lobe was atrophy and left lobe was swelling. Plain computed tomography (CT) showed right lobe was low density. Magnetic resonance (MR) finding was T1-weighted image of right lobe was low intensity and T2-weighted image was high intensity. Angiography showed right lobe was more stained than left lobe. Histologically, right lobe was massive necrosis. These findings suggested that right lobe was liver scar. Biliary imaging showed right lobe was delayed. A 23-year-old-female admitted because of fever and abdominal tumor. Transaminase was normal, only gamma-GTP was elevated. US, plain CT, enhanced CT, MR imaging finding was as same as that of the first case. Similarily, biliary scintigraphy showed right lobe was delayed. Causes of the two liver scars was not clear, whereas liver scar detected after delivery was rare case.

Desensitization for sulfasalazine-induced skin rash in a patient with ulcerative colitis.

A patient with ulcerative colitis developed a sulfasalazine-induced skin allergy manifested by a urticaria rash. The patient underwent drug desensitization. The first desensitization, done according to Holdsworth's protocol, resulted in eruption with itching at a dose of 800 mg. The second desensitization, with Das's protocol, failed to reintroduce the drug because of urticarial eruptions. The third challenge, with a more gradual increase in sulfasalazine dose than that used in Holdsworth's protocol, successfully desensitized the patient. The relationship between the drug and various adverse reactions is reviewed and the desensitization to sulfasalazine is discussed.

[The effects of lowering of blood pressure on pain sensitivity in spontaneously hypertensive rats].

It has been reported that elevation of blood pressure produces a reduction in pain sensitivity. This study was designed to clarify the correlation in spontaneously hypertensive rats (SHR) between fall in blood pressure and pain sensitivity. In seven-weeks-old male SHR, the angiotensin converting inhibitor delapril (10 mg/kg/day) or calcium antagonist nifedipine (3 mg/kg/day) was administered orally every day for 8 weeks. Systolic blood pressure (SBP) pretreatment was significantly higher in the SHR than in normotensive Wistar-Kyoto (WKY) rats and pain sensitivity measured with the hot plate method was significantly lower in the SHR than in the WKY rats. Administration of both drugs produced a significant suppression of elevation of SBP, and produced a significant elevation of pain sensitivity. Furthermore, at 8 weeks after drug administration, urinary norepinephrine (UNE) significantly decreased and plasma beta-endorphin (beta-end) significantly increased. A significant correlation was noted between pain sensitivity and SBP and also between pain sensitivity and UNE. Of these, pain sensitivity was the more closely correlated to degree of change in UNE than to degree of change in SBP. It appears that elevation of pain sensitivity is due to suppression of the sympathetic nervous system by antihypertensive drugs, but not to elevation of beta-end levels. These data suggest that a fall in blood pressure through administration of delapril or nifedipine reverses decrease in pain sensitivity in SHR and that decrease in sympathetic tone plays an important role in the restoration of levels of sensitivity to pain.

Primary ciliary dyskinesia with transverse colon carcinoma.

Primary ciliary dyskinesia, characterized by chronic airway infection, impaired fertility, and situs inversus in half the number of cases, was detected in a 58-year-old Japanese woman, along with carcinoma of the transverse colon. She was of complete situs inversus and subfertile and had chronic bronchitis and nasal polyposis. Ultrastructurally, the cilia of the nasal mucosa lacked dynein arms. This is the first documentation of this disease coupled with cancer.

Adrenal contribution to circulating estrogens in woman.

To study the contribution of adrenal glands to circulating estrogens in woman, the concentrations of estrone (E1), estradiol-17 beta (E2), and estriol (E3) in adrenal and peripheral venous blood were measured by radioimmunoassay and the grandular secretion of estrogens after ACTH stimulation was investigated by analyzing the adrenal vein levels of these steroids in patients with breast cancer who were undergoing a therapeutic adrenal operation. Furthermore, adrenal secretion rates of estrogens and cortisol were estimated. It was shown that there existed greater concentrations of estrogens in adrenal vein than in peripheral blood; about 3 times higher for E1 (p less than 0.001), and 2 times higher for E2 and E3 (p less than 0.05). Administration of ACTH caused a significant increase of E1 and E2 concentrations in adrenal venous blood to mean 150% of the basal levels that was comparable to the increase of cortisol. Apparent adrenal secretion rates of estrogens estimated under surgical situation were calculated to be 7.7 +/- 1.7 (M +/- SE) microgram/day for E1, 1.9 +/- 0.3 microgram/day for E2, and 0.3 +/- 0.2 microgram/day for E3, while the secretion rate of cortisol was 52.7 +/- 8.2 microgram/min. The present study demonstrates that the direct adrenal secretion of not only E1, but E2 and E3 contributes to the circulating estrogen levels, and it is suggested that the adrenal glands might be responsible for the relatively important source of estrogen production in the aged woman.