D-chiro-inositol metabolism in diabetes mellitus.

D-chiro-inositol is a rare inositol isomer present in inositol phosphoglycans which are proposed mediators of insulin action. To study D-chiro-inositol metabolism in diabetes mellitus, a sensitive and specific assay was developed using negative-ion chemical ionization gas chromatography/mass spectrometry. Median urinary D-chiro-inositol excretion, which was 2.1 mumol/day in nondiabetics, was substantially increased to 12 mumol/day in non-insulin-dependent diabetes (P < 0.0001) and to 74 mumol/day in insulin-dependent diabetes (P < 0.0001). Urinary D-chiro-inositol was strongly correlated with fasting plasma glucose (r = 0.568, P < 0.0001), glycated hemoglobin (r = 0.529, P < 0.0001), and urinary glucose (r = 0.368, P = 0.01). The renal clearance of D-chiro-inositol was selectively elevated in both non-insulin-dependent and insulin-dependent diabetes when compared with the clearances of L-chiro-inositol or myo-inositol and exceeded the glomerular filtration rate in 71% of the diabetics but in none of the nondiabetics. In poorly controlled diabetic patients insulin treatment reduced urinary D-chiro-inositol losses by 63% and increased plasma levels by 8.8-fold. The metabolism of D-chiro-inositol is abnormal in diabetes and appears to be influenced by short- and long-term metabolic control.     

The cytoskeleton in Chediak-Higashi syndrome fibroblasts

The Chediak-Higashi syndrome (CHS) trait is expressed in cultured human skin fibroblasts as an abnormal perinuclear concentration of moderately enlarged lysosomes. The cytoskeleton of CHS fibroblasts appears intact. Microtubules are normal in number and morphology, as assessed by colchicine binding studies, antitubulin immunofluorescence, and electron microscopy. Deformability by shear force is unaltered and microfilaments are abundant. However, CHS lysosomes appear to interact abnormally with the cytoskeleton, since the perinculear aggregation partially disperses after depolymerization of cell microtubules with colchicine. These results suggest that CHS is associated with a defect of either the lysosomal membrane itself or of lysosomal membrane- microtubule interaction.     

Somatic complications and contraceptive techniques following legal abortion

The purpose of this study was to establish the frequency of complications following legal abortion at our hospital, and also to examine the use of contraceptives among these patients. All women who requested abortion in the first or second trimester up to week 15 during an 8-month period (n = 1000) were included in the study. The patients were characterized with regard to age, parity, previous abortion, contraceptive technique before and after the abortion, presence of endocervical/urethral Chlamydia trachomatis and frequency and type of somatic complications following the abortion. The overall frequency of complications (infection, bleeding, recurettage) was 5.4%, and 2.7% were readmitted to the hospital. No certain relation between complication rate and gestational length, age or parity was seen, possibly related to selective prostaglandin pretreatment of the cervix and doxycycline treatment of patients with positive cultures for Chlamydia trachomatis. In contrast to previous Scandinavian studies, we found a smaller proportion of young patients, a low percentage of Chlamydial infections, and an unchanged high rate of failure to use contraceptives.     

The cyclin D1 high and cyclin E high subgroups of breast cancer: separate pathways in tumorogenesis based on pattern of genetic aberrations and inactivation of the pRb node

In an attempt to identify subtypes of breast cancer and pinpoint patterns of cell cycle regulatory defects associated with clinical behaviour, proliferation and other transformation associated events, a multitude of cell cycle regulatory proteins were analysed in a material of 113 primary breast cancers. Increased proliferation was observed in two different scenarios; (1) with high cyclin D1 and elevated retinoblastoma protein (pRb) phosphorylation, (cyclin D1(high) tumours) or (2) with high cyclin E protein but low cyclin D1 and lack of corresponding pRb phosphorylation (cyclin E(high) tumours) indicative of an interrupted pRb pathway. Characteristic for cyclin E(high) tumours were further defects in p53, p27 and bcl-2, while c-erbB2 overexpression and c-myc amplification was found in both cyclin D1(high) and E(high) tumours. Using transfected cell lines overexpressing cyclin E, cyclin E(high) and D1(high) tumours were mimicked and the cyclin D1(high) cell line normalized the cyclin E kinase activity by an induction and redirection of p21 and p27 to the cyclin E complex whereas cyclin E(high) cell lines obtained increased kinase activity without redirection of inhibitors. Based on differences in genetic aberrations as well as function of the pRb node we therefore propose a model in which cyclin D1(high) and cyclin E(high) tumours represent two alternative mechanisms to inactivate the pRb pathway and thereby achieve unrestrained growth in the tumorogenesis of breast cancer.     

Phytostanol tablets reduce human LDL-cholesterol

The feasibility of using solid dosage forms containing stanol lecithin to lower human LDL-cholesterol was investigated. The particle size distribution of a coarse aqueous dispersion of a stanol lecithin mixture was determined at various weight ratios of the components. At a stanol-to-lecithin weight ratio of 1.00-1.50, dispersions could be spray dried and the solid reconstituted with water to produce a particle size distribution that was similar to that of the aqueous dispersion from which it was derived. Two solid dosage forms containing this spray-dried stanol lecithin preparation had different disintegration times--tablets less than 10 min and capsules greater than 45 min. Each delivery system was then tested for LDL-cholesterol reduction activity in a placebo-controlled, double-blind clinical trial containing a total of 52 subjects. After a six-week treatment period, the group that received rapidly disintegrating stanol lecithin tablets (1.26 g stanols daily) experienced a decrease in both LDL-cholesterol and the ratio of LDL-cholesterol to HDL-cholesterol by 10.4% (P = 0.01) and 11.5% (P = 0.03), respectively, relative to placebo. On the other hand, with slowly disintegrating capsules (1.01 g daily) there was no statistically significant difference in any lipid parameter between the active group and placebo group. Taken together, these studies demonstrate that for maximum LDL-cholesterol reduction activity the stanol lecithin formulation must be delivered in a rapidly dispersible form to reach the site of cholesterol absorption.     

The role of the dorsomedial striatum in instrumental conditioning

Considerable evidence suggests that, in instrumental conditioning, rats learn the relationship between actions and their specific consequences or outcomes. The present study examined the role of the dorsomedial striatum (DMS) in this type of learning after excitotoxic lesions and reversible, muscimol-induced inactivation. In three experiments, rats were first trained to press two levers for distinct outcomes, and then tested after training using a variety of behavioural assays that have been established to detect action-outcome learning. In Experiment 1, pre-training lesions of the posterior DMS abolished the sensitivity of rats' instrumental performance to both outcome devaluation and contingency degradation when tested in extinction, whereas lesions of the anterior DMS had no effect. In Experiment 2, both pre-training and post-training lesions of the posterior DMS were equally effective in reducing the sensitivity of performance both to devaluation and degradation treatments. In Experiment 3, the infusion of muscimol into the posterior DMS selectively abolished sensitivity of performance to devaluation and contingency degradation without impairing the ability of rats to discriminate either the instrumental actions performed or the identity of the earned outcomes. Taken together, these results suggest that the posterior region of the DMS is a crucial neural substrate for the acquisition and expression of action-outcome associations in instrumental conditioning.     

Resistive training and chromium picolinate: effects on inositols and liver and kidney functions in older adults

This study assessed the effects of resistive training (RT) with or without chromium picolinate (Cr-pic) supplementation on the 24-h urinary excretions of myo-inositol, D-chiro-inositol, and pinitol, as well as clinical indices of kidney and liver functions. Thirty-two nondiabetic subjects, age 62 +/- 4 y, performed RT twice weekly for 12 wk and consumed either 924 ug Cr/d as Cr-pic (n = 17) or a placebo (n = 15). Whole-body strength increased in all subjects by 20 % and urinary chromium excretion increased 47-fold in the Cr-pic group. Urinary myo-inositol, D-chiro-inositol, and pinitol were not changed with RT or influenced by Cr-pic. Serum indices of kidney and liver functions were within clinically normal ranges at baseline and the end of the study. These results suggest that RT did not influence the urinary excretions of inositols. High dose Cr-pic did not influence the urinary excretion of inositols and the selected indices of kidney and liver functions in conjunction with RT.     

Decreased expression of thioredoxin and glutaredoxin in placentae from pregnancies with pre-eclampsia and intrauterine growth restriction

Pre-eclampsia is one of the major contributors to perinatal morbidity. This study was performed to test a hypothesis which suggests that pre-eclampsia is associated with inadequate control by the thioredoxin system and other related reducing systems. Placental tissue from normal pregnancies (NC), severe pre-eclampsia with fetuses small for gestational age (SPE), mild pre-eclampsia with fetuses small for gestational age (MPE) and pregnancies with small fetuses for gestational age without any sign of pre-eclampsia (IUGR) was collected immediately after delivery. The mRNA levels for thioredoxin and glutaredoxin were determined using a solution hybridization method and the distribution of the proteins in a normal placenta was analysed by immunohistochemistry. Results showed that the thioredoxin mRNA level in the SPE group was decreased to one third of the level in the NC group. Also the IUGR group showed a significant decrease. The glutaredoxin mRNA level in the SPE group was one half of that seen in the NC group. There was significant correlation between the mRNA levels for thioredoxin and glutaredoxin, both in the normal and growth restricted pregnancies. We conclude that the thioredoxin and glutaredoxin reducing systems are affected in placenta from pregnancies with pre-eclampsia and/or growth restriction of fetuses, and that the decrease correlates to the severity of the condition.     

Postoperative comfort in paediatric outpatient surgery.

Postoperative conditions in hospital and at home were evaluated in 200 paediatric daycase patients by using questionnaires and telephone interviews. Pain was assessed by behaviour observation or a faces rating scale depending on age. Anaesthetic methods, nausea/vomiting, analgesics and parents' aspects were also recorded. Seventy per cent of the patients received regional anaesthesia. Immediate postoperative analgesia was satisfactory in 75% of the children. When the effects of intraoperatively administered analgesics wore off at home almost half the children rated higher than mild pain. The increased degree of pain at home was especially pronounced after regional anaesthesia. The total incidence of nausea/vomiting was 28% and fentanyl caused nausea and vomiting in a significantly higher proportion of cases. The study points out that immediate postoperative comfort obtained by prophylactic analgesia needs to be followed by analgesics given on a continuous basis for the first days after surgery.     

Immunosorbent Chemistry: A Study of Agarose-Based Column Sorbents for the Removal of Low-Density Lipoprotein (LDL) from Blood

Anti-LDL antibody was covalently attached to agarose beads in order to prepare an immunosorbent for the removal of LDL from plasma or blood. Both the conditions of antibody coupling and the type of agarose matrix used were critical to the optimization of LDL binding capacity. Sorbents binding 6-8 mg lipoprotein cholesterol/ml column volume were obtained using cyanogen bromide or glutaraldehyde coupling procedures and crosslinked 2% agarose beads. The sorbent could be regenerated by washing with 1 M acetic acid, a reagent that was also an effective disinfectant. In vitro perfusions of whole blood over small columns of 212-300 mu beads showed excellent flow rates (2 ml/min/cm2 under 50-100 cm saline pressure); recovery of leukocytes and platelets exceeded 90%, and complement was not activated. Leakage of antibody and bead matrix was negligible. The antibody-agarose beads could not be sterilized by conventional techniques, but withstood treatment with 0.34% phosphoric acid in 80% ethanol at 37 degrees C, a novel method of chemical sterilization.