Application of carboxymethyl-beta-cyclodextrin as a chiral selector in capillary electrophoresis for enantiomer separation of selected neurotransmitters.

The aim of this work was to optimize conditions for capillary electrophoresis separation of different neurotransmitters (serotonin, phenylalanine, dopamine, adrenaline, ephedrine, propranolol and DOPA) in a single run, including separation of existing enantiomers. As chiral selectors added to the borate background, electrolyte unsubstituted alpha-, beta- and -gamma-cyclodextrins (CDs), methyl-, dimethyl-, and trimethyl-substituted beta-CDs, and hydroxypropyl-substituted alpha-, beta- and gamma-CDs were examined. Also carboxymethyl-beta-CD and succinyl-beta-CD were used for this purpose. In addition to the kind and concentration of chiral selector, some other experimental factors also have been optimized, such as concentration of borate buffer, content of methanol, pH of electrolyte, method of sample introduction into the capillary and washing procedure between consecutive runs. The best results were obtained using 20 mM carboxymethyl-beta-CD in borate buffer of pH 7.5 as running electrolyte and hydrostatic injection. The obtained sensitivity of response (peak height) varied from 0.4 for adrenalines to 2.3 mAU mM(-1) for propranolols. The concentration detection limits (S/N=3) were in the range from 0.04 mM for propranolols to 0.2 mM for adrenalines. The resolution obtained in optimized conditions in a single run was from 0.75 for adrenalins and 1.0 for propranolols up to 2.0 for ephedrines. The developed method was employed for determination of these analytes in brain tissue extracts.

[Microalbuminuria as a risk factor for diabetic osteopathy in patients with IDDM and renal sufficiency]. / Mikroalbuminuria jako czynnik ryzyka osteopatii cukrzycowej u pacjentów z IDDM w okresie wydolnosci nerek.

Disturbances in bone marrow vascularisation can be one of the causes of diabetic osteopathy. The aim of the study was to answers the question if microalbuminuria as a results of capillary injury can be a sign of bone mineralisation disorders in IDDM renal sufficient patients. We examined 60 IDDM patients (30 women without menstruation disturbances; 30 men; age 25-36 years old). All the observed subjects were divided into groups: I-30 normoalbuminuric patients (0-29 mg/24 h); II-30 microabuminuric patients (30-295 mg/24 h). Bone mineral density (BMD) of femoral neck, lumbar spine (L2-L4) and total body was measured by dual energy X-ray absorptiometry (DEXA, Lunar). The biochemical parameters of bone turnover were measured both in serum and urine as follows: osteocalcine, total hydroxyproline (HPR, HPR/Cr), total alkaline phosphatase (AP) with bone fraction, total calcium (Ca, Ca/Cr) and inorganic phosphor (P). Microalbuminuric patients presented more severe bone turnover disturbances, shown by differences in: BMD and Z-score for femoral neck (p < 0.05), serum HPR (p < 0.05), AP (p < 0.05), AP (p < 0.01) and its bone fraction (p < 0.05). We proved the presence of statistically significant correlation coefficients for albuminuria and some densytometric and biochemical bone parameeters. Our results suggest that microalbuminuria can indirectly indicate the dynamic of bone turnover derangement in IDDM course. They are present mostly in the femoral neck, which because of the vascularisation type is particularly susceptible to subalimentation in the diabetic microangiopathy course.

[Bone mineralization in insulin-dependent diabetes mellitus]. / Mineralizacja tkanki kostnej w cukrzycy insulinozaleznej.

One of insulin-dependent diabetes mellitus (IDDM) complications are bone mineral disorders called diabetic osteopathy. Because of many controversies of this subject we took a research of bone mineralisation in IDDM patients before 40 years old. The evaluation of BMD was performed with the use of X-ray dual energy absorptiometry--DEXA in the AP projection for lumbar part of vertebral column (BMD L2-L4), left femur neck (BMD-neck) and total skeleton (BMD-total). Bone tissue metabolism was evaluated with the aid of biochemical tests. The examined group consisted of 99 patients with IDDM (45 women and 54 men), without any other risk factors for changes in bone metabolism. The results were related to: sex, age in which IDDM was diagnosed, duration of IDDM, metabolic control of diabetes and to some IDDM complications. The control group consisted of 113 healthy subjects matched for age, sex, weight, height and calcium diet. We observed that BMD in IDDM patients before 40 years old was significantly lower than in healthy subject. One of some diabetic osteopathy pathomechanism seems to be an advantage of bone resorption over bone formation. Bone demineralisation which was observed to be most pronounced in femur neck, was not related to age, differed in accordance to sex and was higher of age when IDDM appeared was lower. BMD was also related to duration of IDDM and metabolic control of IDDM.

[Effect of low-dose recombinant humane erythropoietin therapy on the quality of life in patients with anemia in the course of end-stage renal failure treated with dialysis]. / Wplyw malych dawek ludzkiej rekombinowanej erytropoetyny na jakosc zycia chorych z niedokrwistoscia w przebiegu schylkowej niewydolnosci nerek leczonych dializami pozaustrojowymi.

The aim of the study was to assess the efficacy of low-dose subcutaneous recombinant human erythropoietin (rHuEpo) therapy in hemodialysis patients with particular emphasis on their quality of life. Twenty five anemic (Ht25%) patients (14 males and 11 females, age 39-13 years) with end-stage renal disease were given rHuEpo (initial dose: 52.5 +/- 2.5 IU/kg/week; maintenance dose: 67.0-10.5 IU/kg/week) once or twice weekly for 12 months. Quality of life, assessed by self-administered questionnaire (1-3 scale), was measured every month. Additionally, sexual functions (-1 up to 3 scale, basal level 0), including libido and sexual satisfaction, and serum sex hormones (testosterone, LH, FSH, prolactin) were evaluated every 6 months. During first 4 months of the therapy there was a significant increase of Ht (21.1 +/- 0.5% vs 28.5 +/- 0.6%; p < 0.0001), which was maintained for the whole study period. From the 3rd month in majority of patients a marked (p < 0.01) improvement in their physical fitness, mood and cold tolerance was noted. Despite a substantial increase in sexual satisfaction (p < 0.01) and libido (p < 0.001), no significant changes in serum sex hormones profile, except transient rise in serum prolactin level, were observed. It is concluded that low-dose rHuEpo therapy for the renal anemia of hemodialysis patients is associated with a sustained significant improvement in their quality of life and sexual functions, despite no significant changes in sex hormones serum levels.

[Combined therapy with calcitonin and high doses of active vitamin D3 metabolites in uremic hyperparathyroidism]. / Skojarzone leczenie kalcytonina i duzymi dawkami aktywnych metabolitów witaminy D3 we wtórnej do mocznicy nadczynnosci przytarczyc.

Active vitamin D3 pulse therapy effectively suppresses parathormone (PTH) synthesis in uremic hyperparathyroidism but high serum levels of calcitriol achieved can induce direct osteoclastic resorption and block bone formation. Therefore we found it interesting to examine whether an addition of the osteoclast inhibitor, calcitonin (CT), could reduce those unwanted effects. 75 hemodialysis patients with at least 5-fold 1-84 PTH serum level elevation were divided into 4 treatment groups: I (n = 19)-CT and 1 alpha-OH-D3; II (n = 20)-CT; III (n = 19)-1 alpha-OH-D3 (n = 10) or 1.25 (OH)2D3 (n = 9) alone; IV (n = 17)-none of these drugs. CT (200 IU) and 1 alpha-OH-D3/1.25(OH)2D3 (up to 5 micrograms) were given 3 times a week. Dialysate Ca was 1.40-1.45 (Group I, III) or 1.95-2.00 mmol/l (Group II, IV). Within 8 months serum 1-84 PTH fell by 75% (p < 0.001) in Group I and by 77% (p < 0.001) in Group II, serum Ca increased by 0.22 +/- 0.05 mmol/l in Group I (p < 0.005) and by 0.25 +/- 0.05 mmol/l in Group III (p < 0.005), alkaline phosphatase activity decreased by 35% in Group I (p < 0.01) and 31% in Group III (p < 0.005) whereas in Groups II and IV no significant changes were noted. In Group III no differences between patients taking 1 alpha-OH-D3 or 1.25 (OH)2D3 were observed. The significant reduction of serum hydroxyproline (37%; p < 0.001) was seen only in Group 1. The increase in bone mineral density (BMD) measured by dual-energy X-ray absorptiometry was greater in Group I than in Group III (p < 0.05). In Group II the effect was mostly insignificant, whereas in Group IV a substantial decrease (p < 0.001) in BMD was observed. These data suggest that combined therapy with CT and oral 1 alpha-OH-D3 pulses is more effective than pulses alone in inhibiting bone resorption and in increasing BMD in hemodialysis patients with uremic hyperparathyroid bone disease.

[Evaluation of treating anemia in patients undergoing chronic dialysis with small doses of human recombinant erythropoietin]. / Ocena leczenia niedokrwistosci u chorych przewlekle dializowanych malymi dawkami ludzkiej rekombinowanej erytropoetyny.

The aim of the study was to assess the efficacy of low dose subcutaneous (sc) recombinant human erythropoietin (rHuEpo) therapy in hemodialysis (hd) patients, with particular emphasis on their quality of life. 25 anemic (Ht < 25%) hd patients with end-stage renal disease were given small sc doses of rHuEpo once or twice weekly for 12 months. During first 4 months of the therapy there was a significant increase of Ht (21.1 +/- 0.5 vs 28.5 +/- 0.6%; p < 0.0001) and serum hb (6.68 +/- 0.12 vs 8.51 +/- 0.18 g/dl; p < 0.0001) at mean induction dose of 52.5 +/- 2.5 IU/kg/week and this was maintained with a mean dose 67.0 +/- 10.5 IU/kg/week. RHuEpo was effective in 24 patients; all of them required no blood transfusions after starting the therapy. In majority of patients an substantial (p < 0.01) improvements in exercise tolerance, well-being, cold tolerance, sexual satisfaction and libido were observed, although sexual hormones profile revealed no significant changes during the treatment. Within first 6 months of the study cardiac index decreased substantially (p < 0.01), mainly because of stroke volume reduction, but after one year this hemodynamic improvement was noted only in patients who maintained a stable blood pressure. Hypertension worsened in 31% of patients. Low-dose s.c. rHuEpo: (1) is effective and safe treatment for anemia in hds patients, sufficient to abolish blood transfusion requirements; (2) produces significant improvements in quality of life; and (3) allows for 50% costs reduction.