Effects of Storage Temperature and Time on Stability of Serum Tacrolimus and Cyclosporine A Levels in Whole Blood by LC-MS/MS.

Tacrolimus and cyclosporine A are immunosuppressant drugs with narrow therapeutic windows. The aim of this study was to investigate the stability of tacrolimus and cyclosporin A levels in whole blood samples under different storage conditions. Whole blood samples were obtained from 15 patients receiving tacrolimus and 15 patients receiving cyclosporine A. Samples were immediately analyzed and then stored at different conditions (room temperature (24°C-26°C) for 24 hours, +4°C for 24 and 48 hours, and -20°C for one month) and then analyzed again. For tacrolimus, there was a significant difference between samples analyzed immediately and those kept 24 hours at room temperature (P = 0.005) (percent change 32.89%). However, there were no significant differences between the other groups. For cyclosporine A, there was a significant difference between samples analyzed immediately and those kept 24 hours (P = 0.003) (percent change 19.47%) and 48 hours (P = 0.002) (percent change 15.38%) at +4°C and those kept 24 hours at room temperature (P = 0.011) (percent change 9.71%). Samples of tacrolimus should be analyzed immediately or stored at either +4°C or -20°C, while samples of cyclosporine A should be analyzed immediately or stored at -20°C.

Serum levels of carcinoembryonic antigen (CEA) in patients with bipolar disorder.

OBJECTIVE: Carcinoembryonic antigen (CEA) is an oncofetal glycoprotein that is widely used as a tumour marker in adenocarcinomas. However, several non-neoplastic conditions, including acute and chronic inflammation and other inflammation-related conditions, are characterised by increased CEA concentrations. Bipolar disorder (BD) ranks seventh among the worldwide burden of non-fatal diseases. Inflammatory biomarkers have been considered as one of the main key pillars of a multifactorial approach for prediction of BD in an at-risk population. BP is accompanied by activation of inflammatory, cell-mediated and negative immunoregulatory cytokines. METHODS: We measured the levels of CEA in serum samples from 44 individuals with euthymic BP out-patients and 45 healthy controls. Patients were diagnosed according to the DSM-IV criteria. CEA was measured by an electrochemiluminescence immunoassay. RESULTS: The mean serum CEA concentration was 2.36±1.52 and 1.77±0.98 µg/l in patients and controls, respectively. CEA levels were significantly increased in euthymic BP patients when compared with controls (p=0.031). CONCLUSIONS: This study suggests that CEA is increased in BD and supports a role for immune activation in the core pathological mechanisms of BP. CEA levels may be a secondary marker for diagnosing BP.

Effect of pneumatic tube delivery system rate and distance on hemolysis of blood specimens.

BACKGROUND: We evaluated the effects of pneumatic tube system (PTS) transport rates and distances on routine hematology and coagulation analysis. PTS effects on centrifuged blood samples were also examined. METHOD: The study was completed at Dicle University Hospital, which has the longest pneumatic tube system in Turkey. Blood samples were collected at three different locations within the hospital and an emergency department, and delivered to the central laboratory by the PTS or a human carrier. Samples were transported at different rates and over varying distances. Each specimen's potassium (K) and lactic dehydrogenase (LDH) levels, in both the serum and plasma, were tracked to monitor hemolysis. Measurements of LDH and K were obtained using heparin or citrate. RESULT: A positive correlation was observed between distance and hemolysis in serum samples transported at 4.2 m/sec, and at 3.1 m/sec for more than 2200 m (r = 0.774 and r = 0.766, respectively). Distance and hemolysis were also correlated in non-centrifuged samples (r = 0.871). The alterations in plasma LDH and K levels at different rates and PTS lengths were not statistically significant. CONCLUSION: The rate of hemolysis in PTS transported samples, dependent on PTS length and rate, may seriously affect routine tests of non-centrifuged samples.

Routine enzymes in the monitoring of type 2 diabetes mellitus.

We examined the relationships of glucose and HbA1c levels with the routinely screened serum enzyme activities in type 2 diabetes mellitus, and we designed an in vitro study to evaluate the direct effect of glucose levels on enzyme activities. The study was performed on a consecutive series of outpatients with type 2 diabetes who were followed up at Dicle University Medical Faculty Hospital from May 2009 to May 2010 for the first time. Effects of aspartate transaminase, aminotransferase, gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) activities, glucose and HbA1c levels and in vitro glucose (492, 287, 184, 131, 82 mg dl⁻¹, respectively) on enzymes were determined. The patients were categorized on the basis of glucose and HbA1c levels and grouped according to a range of values. In patients with high HbA1c levels (>10.1%), ALP, GGT activities and creatine kinase (CK)-MB/CK (p = 0.008, 0.026, 0.014) ratio were increased significantly when compared with those in the control group. In patients with high glucose levels (> 200 mg dl⁻¹), ALP, GGT activities and CK-MB/CK ratio (p = 0.003, 0.001, 0.001) were increased significantly when compared with those in the control group. Glucose, which was added to serum in different concentrations in vitro, did not directly affect enzyme activities such as ALP, GGT and CK. We concluded that increased glucose levels could damage the liver and the heart muscle cells. Monitoring of blood glucose levels is a more valuable parameter than monitoring HbA1c in the momentary evaluation of diabetes.

The effect of calcitonin on biochemical markers and zinc excretion in postmenopausal osteoporosis.

OBJECTIVES: A double-blind, randomized, placebo-controlled trial was conducted in women with postmenopausal osteoporosis to evaluate effects on biochemical markers and urinary excretion of zinc (Zn) of calcitonin therapy. METHODS: Patients were required to have a bone mineral density (BMD) of 2.5 S.D. or more below the young adult mean either at the postero-anterior lumbar spine or at the femoral neck. Subjects were eligible for our study if they were 50 years or older, with at least 5 years of menopause, and in good general health as determined by medical history and a routine clinical blood analysis. The patients were randomly assigned to receive intranasal salmon calcitonin (200 IU/day; 50 patients) or placebo (50 patients). All patients received supplemental calcium (1000 mg/day). Additionally, 40 age-matched, demographically similar, healthy postmenopausal women were also selected as controls. Measurements of cross-linked N-telopeptides of type I collagen (uNTx), osteocalcin (sOC), and urinary zinc concentration were done. All parameters were measured before therapy and again after 1, 3 and 6 months. RESULTS: After 3 and 6 months of treatment, a higher decrease of most indices was observed in the calcitonin group. A statistically significant decrease occurred in the levels of sOC, uNTx and uZn after 3 and 6 months in patients receiving calcitonin therapy (P<0.05). Levels of sOC and uNTx in calcitonin group were significantly different after 3 and 6 months from both placebo and baseline values of calcitonin group (P<0.05). Levels of sOC, uNTx and uZn decreased about 40, 46 and 37%, respectively, in calcitonin group at 6 months after the start of treatment. CONCLUSIONS: Our study suggests that values of uNTx, uZn and sOC were significantly lower in the patient group than those in control group and in postmenopausal women with osteopororsis, calcitonin reduces the concentrations of uNTx, uZn and sOC.

Comparison of zinc excretion and biochemical markers of bone remodelling in the assessment of the effects of alendronate and calcitonin on bone in postmenopausal osteoporosis.

OBJECTIVES: The aim of this study was to determinate the clinical usefulness of urinary bone resorption markers, urinary zinc excretion, and other biochemical markers in postmenopausal women with osteoporosis. We also evaluated the effects of alendronate and calcitonin therapies on biochemical markers of bone remodeling and urinary zinc excretion over a 6-month period. SUBJECTS AND METHODS: The study design was a randomized placebo controlled study. The patients were randomly assigned to receive alendronate (10 mg/day; 45 patients) or calcitonin (200 IU/day; 45 patients) or placebo (45 patients) for 6 months. All patients received supplemental calcium (1000 mg/day). To assess bone resorption, we measured excretion of cross-linked N-telopeptides of Type I collagen (uNTx); urinary zinc concentrations and standard chemistry determinations were also measured; and osteocalcin (sOC) was measured as a marker of bone formation. All parameters were measured before therapy and again after 1, 3, and 6 months in all groups. RESULTS: A statistically significant decrease occurred in the levels of sOC, uZn, and uNTx after 3 and 6 months in patients receiving calcitonin therapy (P < 0.05). sOC, uZn, and uNTx levels in the calcitonin group were significantly lower after three and 6 months from both placebo and baseline values of calcitonin group. In the alendronate group, a statistically significant decrease was observed in the levels of uNTx and uZn after 1 month and in the sOC, uZn, and uNTx after 3 and 6 months from both placebo and baseline values of alendronate group (P < 0.05). uNTx, sOC, and uZn decreased about 44%, 36%, and 33%, respectively, in the calcitonin group and about 53%, 51%, and 38%, respectively, in the alendronate group below baseline values 6 months after initiating treatment. In the placebo group, there was no significant decrease in sOC, uZn, and uNTx during the course of the study. CONCLUSION: Our study suggests that in postmenopausal women with osteoporosis, both alendronate and calcitonin reduce the concentrations of uNTx, uZn, and sOC, the effect of the alendronate dose administered being significantly earlier and more pronounced. Measurement of uNTx, uZn, and sOC might be a useful biochemical method of observing the positive clinical effect following alendronate or calcitonin therapy in postmenopausal women.

Cortisol and hypothalamic-pituitary-gonadal axis hormones in follicular-phase women with fibromyalgia and chronic fatigue syndrome and effect of depressive symptoms on these hormones.

We investigated abnormalities of the hypothalamic-pituitary-gonadal axis and cortisol concentrations in women with fibromyalgia and chronic fatigue syndrome (CFS) who were in the follicular phase of their menstrual cycle, and whether their scores for depressive symptoms were related to levels of these hormones. A total of 176 subjects participated - 46 healthy volunteers, 68 patients with fibromyalgia, and 62 patients with CFS. We examined concentrations of follicle-stimulating hormone, luteinizing hormone (LH), estradiol, progesterone, prolactin, and cortisol. Depressive symptoms were assessed using the Beck Depression Inventory (BDI). Cortisol levels were significantly lower in patients with fibromyalgia or CFS than in healthy controls (P < 0.05); there were no significant differences in other hormone levels between the three groups. Fibromyalgia patients with high BDI scores had significantly lower cortisol levels than controls (P < 0.05), and so did CFS patients, regardless of their BDI scores (P < 0.05). Among patients without depressive symptoms, cortisol levels were lower in CFS than in fibromyalgia (P < 0.05). Our study suggests that in spite of low morning cortisol concentrations, the only abnormalities in hypothalamic-pituitary-gonadal axis hormones among follicular-phase women with fibromyalgia or CFS are those of LH levels in fibromyalgia patients with a low BDI score. Depression may lower cortisol and LH levels, or, alternatively, low morning cortisol may be a biological factor that contributes to depressive symptoms in fibromyalgia. These parameters therefore must be taken into account in future investigations.

The role of trace minerals in the pathogenesis of postmenopausal osteoporosis and a new effect of calcitonin.

The physiologic role of calcitonin in mineral and bone homeostasis is not very well understood. Very few longitudinal studies have reported the effects of calcitonin therapy on trace minerals in postmenopausal osteoporosis despite the documented involvement of trace minerals in normal skeletal metabolism. Several trace minerals, particularly magnesium (Mg) and zinc (Zn), essential for organic bone matrix synthesis have been known for at least three decades. The present study was designed to determine whether the mineral profile was different between 70 osteoporotic and 30 nonosteoporotic postmenopausal women and to evaluate the efficacy of calcitonin therapy for 6 months on these trace minerals in postmenopausal osteoporotic women. In our study, the serum values of Mg, copper (Cu), and Zn (P < 0.05) were significantly lower in the patient group than those in the control group. After 3 months of treatment, serum Cu, Zn, and Mg levels did not differ between the patients and controls, and this situation has continued after the end of 6 months of therapy. Serum Cu, Zn, and Mg levels increased consistently during the 6-month treatment period. The higher levels of serum Mg in the 3rd and 6th months of therapy were found to be statistically significant compared to those before treatment (P < 0.05). Serum Cu and Zn levels were found to be significantly higher at all measurements during the treatment period as well as at the end of therapy (P < 0.05). These results suggest that (1) calcitonin therapy regulates Mg, Cu, and Zn levels in postmenopausal osteoporosis; (2) when serum calcium and phosphorus were normal in postmenopausal osteoporosis, serum Mg, Cu, and Zn were more useful for evaluation; and (3) further studies are essential to evaluate the role of dietary composition on the manifestations of osteoporosis.