Reduced glucose tolerance and insulin resistance induced by steroid treatment, relative physical inactivity, and high-calorie diet impairs the incretin effect in healthy subjects.

AIMS/HYPOTHESIS: The loss of incretin effect in patients with type 2 diabetes mellitus may be secondary to impaired glucose homeostasis. We investigated whether reduced glucose tolerance and insulin resistance induced by steroid treatment, relative physical inactivity, and high-calorie diet in healthy young males would impair the incretin effect. METHODS: The incretin effect was measured using 75 g oral glucose tolerance test (OGTT) and isoglycemic iv glucose infusion (IIGI) in 10 healthy Caucasian normal glucose-tolerant male subjects without any family history of diabetes [age 24 + or - 3 yr (mean + or - sd); body mass index 23 + or - 2 kg/m(2); glycosylated hemoglobin 5.4 + or - 0.1%] before and at the end of a 12-d period with oral administration of prednisolone (37.5 mg once daily), high-calorie diet, and relative physical inactivity. RESULTS: The 12-d intervention period resulted in significant increases in body weight [79 + or - 5 vs. 80 + or - 6 kg (mean + or - sd), P = 0.03] and fasting plasma glucose (5.1 + or - 0.1 vs. 5.6 + or - 0.2 mm, P = 0.016), whereas insulin sensitivity (Matsuda index 17.6 + or - 1.7 vs. 9.2 + or - 1.0, P = 0.0001) decreased. Glucose tolerance [as assessed by the 120-min plasma glucose value after OGTT (4.9 + or - 1.1 vs. 7.8 + or - 2.5 mm, P < 0.0001) and area under curve (AUC) (152 + or - 45 vs. 384 + or - 53 mm.4 h, P = 0.002)] during the OGTT deteriorated. Also, the incretin effect [incretin effect (percent) = 100% x (AUC(insulin,OGTT) - AUC(insulin,IIGI))/AUC(insulin,OGTT))] deteriorated (72 + or - 5 vs. 43 + or - 7%, P = 0.002). An increase in glucose-dependent insulinotropic polypeptide response during OGTT, but no significant changes in glucagon-like peptide-1 or glucagon responses, was observed after glucose homeostatic dysregulation. CONCLUSIONS/INTERPRETATION: Impairment of the incretin effect can be elicited by a short period of reduced glucose tolerance and insulin resistance in healthy male subjects not disposed for type 2 diabetes.

Treatment of type 2 diabetes with glucagon-like peptide-1 receptor agonists.

The incretin system is an area of great interest for the development of new therapies for the management of type 2 diabetes. Existing antidiabetic drugs are often insufficient at getting patients to glycaemic goals. Furthermore, current treatment modalities are not able to prevent the continued ongoing decline in pancreatic beta-cell function and, lastly, they have a number of side effects including hypoglycaemia and weight gain. Glucagon-like peptide-1 (GLP-1) receptor agonists are a new class of pharmacological agents, which improve glucose homeostasis in a multifaceted way. Their effects include potentiation of glucose-stimulated insulin secretion, glucose-dependent inhibition of glucagon secretion and reduction in gastric emptying, appetite, food intake and body weight. Additionally, preclinical data suggest that they may preserve beta-cell mass and function. The incidence of hypoglycaemia with GLP-1 receptor agonists is low, the compounds have clinically relevant effects on body weight, and data are suggesting beneficial effects on cardiovascular risk factors. Exenatide was released in 2005 for the treatment of type 2 diabetes and liraglutide is expected to be approved by the Food and Drug Administration in US and the European Medical Agency in Europe for use in 2009. In this review, the available data on the two drugs are presented and discussed.

Pharmacological characterization of mouse GPRC6A, an L-alpha-amino-acid receptor modulated by divalent cations.

BACKGROUND AND PURPOSE: GPRC6A is a novel member of family C of G protein-coupled receptors with so far unknown function. We have recently described both human and mouse GPRC6A as receptors for L-alpha-amino acids. To date, functional characterization of wild-type GPRC6A has been impaired by the lack of activity in quantitative functional assays. The aim of this study was thus to develop such an assay and extend the pharmacological characterization of GPRC6A. EXPERIMENTAL APPROACH: We have engineered a novel cell-based inositol phosphate turnover assay for wild-type mouse GPRC6A based on transient co-expression with the promiscuous Galpha(qG66D) protein, known to increase receptor signalling sensitivity. This assay allowed for measurements of L-alpha-amino acid potencies. Furthermore, in combination with an assay measuring inward currents at Ca(2+)-activated chloride channels in Xenopus oocytes, the divalent cation-sensing ability of the receptor was examined. KEY RESULTS: Using our novel assay, we demonstrate that the basic L-alpha-amino acids ornithine, lysine, and arginine are the most potent agonists at wild-type mouse GPRC6A. Using two different assay systems, we show that divalent cations do not activate the G(q) signalling pathway of mouse GPRC6A per se but positively modulate the amino-acid response. CONCLUSIONS AND IMPLICATIONS: This is the first reported assay for a wild-type GPRC6A successfully applied for quantitative pharmacological characterization of amino acid and divalent cation responses at mouse GPRC6A. The assay enables further search for GPRC6A ligands such as allosteric modulators, which may provide essential information about the physiological function of GPRC6A.

Absence of high-affinity calreticulin autoantibodies in patients with systemic rheumatic diseases and coeliac disease.

Calreticulin has been reported to be an autoantigen in various autoimmune connective tissue diseases and in coeliac disease. Previous studies have used incubation buffers with low salt and low detergent concentrations (low stringency conditions) with serum albumin or other proteins as a blocking agent. Using these conditions we found a relatively high level of non-specific binding in many sera. Antibodies to proteins that are used as blocking reagents in ELISA (bovine serum albumin (BSA), ovalbumin, skimmed milk powder) are frequently present in sera, and these may cause false-positive results. Moreover, the low isoelectric point of calreticulin and its chaperone properties may give rise to false-positive results under low stringency conditions. We report that the use of a simple buffer without protein (50 mM Tris, pH 7.5, 1% Tween 20, 0.3 M NaCl) removes most of the problems with unwanted binding (high stringency conditions). Using the high stringency conditions, we screened sera from 107 patients with systemic lupus erythematosus, sera from patients with other systemic autoimmune diseases and from children with coeliac disease for the presence of high-affinity calreticulin autoantibodies by immunoblotting and ELISA. None of the sera contained high-affinity calreticulin antibodies. It is concluded that calreticulin is not a common autoantigen in patients with autoimmune connective tissue diseases or coeliac disease.

Energy intake and milk production in mink (Mustela vison)--effect of litter size.

Energy intake and milk production were measured in 12 mink dams raising litters of 3, 6 and 9 kits one to four weeks post partum by means of balance experiments and measurements of milk intake of the kits by the water isotope dilution technique. The dams were fed ad libitum on a conventional wet mink diet (DM: 323 g/kg; CP: 173 g/kg; ME: 4.4 MJ/kg). Milk samples collected from dams with corresponding litter sizes and lactation weeks, and body composition of kits nursed by these dams, were analysed for content of DM, ash, N and fat. The ME and drinking water consumption were higher in dams nursing 9 kits than in dams nursing 3 kits. The N and water balances as well as the live weight of dams were not affected by litter size. Daily milk production was higher in dams nursing 9 kits than in dams nursing 3 kits. The DM, N and fat content of the milk increased during lactation, but were not affected by litter size. Individual kit live weight was higher in litters of 3 than in litters of 6 and 9 kits four weeks post partum. The DM and fat content of the kits were lowest in kits from litters of 9 kits, whereas these kits had the highest protein content. Daily ME for maintenance of kits and the efficiency of utilisation of ME in milk for body gain were estimated to 356 kJ/kg0.75, kp approximately 0.53 and kf approximately 0.71, respectively. In conclusion, daily milk production increased with increasing litter size, but not in proportion to the number of kits, indicating that milk production limits the growth rate of the young. In the fourth week of lactation, milk production was not different between dams nursing 6 or 9 kits, indicating a maximum capacity.

Effect of supplementation with the antioxidant astaxanthin on reproduction, pre-weaning growth performance of kits and daily milk intake in mink.

The study comprised two parts. Firstly, the effects of dietary supplementation with an algal meal (Novasta) with a high astaxanthin content on ovulation rate (number of corpora lutea, implantation rate, number, mass and length of fetuses) of breeding female mink were evaluated. Secondly, reproductive outcome (number of live and stillborn kits), kit growth rate and milk intake were studied. Both studies were performed on standard brown female mink (n = 20; control (n = 10) and experimental (n = 10)) housed under conventional farm conditions. Experimental animals were supplied with 5.35 mg astaxanthin per day (0.25 g algal meal (Novasta)). The numbers of corpora lutea, implantation sites and fetuses appeared to be higher in the group that was given astaxanthin but the effect was not significant. The differences between treated and control mink were 1.4 (corpora lutea), 0.9 (implantation sites) and 1.2 (litter size). The percentage of stillborn kits was reduced by 6.3 (P < 0.005). The milk intake as measured by use of the isotopic water dilution technique was not affected by treatment group. Milk intake increased from about 19 g in week 1 of lactation to about 30 g per kit per day in week 4 of lactation. Kit weight gain was not affected by the experimental treatment.

[Acute myeloid leukemia. Therapeutic results 1985-1994]. / Akut myeloid leukaemi. Behandlingsresultater 1985-1994.

In a retrospective study we evaluate the treatment and outcome of 421 adults admitted to our department with acute myeloid leukemia (AML) during the 10 year period from 1985-1994. Younger patients (< or = 55 years) had a significantly better prognosis than elderly patients (> 55 years), partly because more younger patients had remission-induction therapy (81% versus 39%) and their complete remission (CR) rate was higher (69% versus 41%). In patients achieving CR the long-term survival (five years) was 40% for younger and 26% for elderly patients. Nineteen patients received autologous bone marrow transplantation (BMT) and eight an allogeneic BMT in first CR. Five patients treated with allogeneic BMT are still relapse-free, whereas autologous BMT, in comparison to conventional chemotherapy alone, did not improve the five-year relapse-free survival (29 versus 27%). Our data illustrate that in a non-selected material of AML patients the long-term survival (five years) has only slightly improved. The effect of autologous BMT cannot be evaluated in this retrospective setting.

Anticoagulant properties of rabbit lungs in tissue thromboplastin-induced intravascular coagulation.

This study was conducted in order to examine possible anticoagulant properties of the lungs during tissue thromboplastin-induced intravascular coagulation. Rabbit brain tissue thromboplastin (n = 17) or saline (n = 6 + 3) was infused above the right atrium (n = 11 + 3) of the heart or in the arcus aorta (n = 6) for a period of 120 min in non-pregnant New Zealand rabbits. Rabbits infused with tissue thromboplastin responded with significantly (p < 0.05) more excessive changes in a number of haemodynamic variables (heart rate, PaO2,PaCO2, blood pH etc.) compared with rabbits infused with saline. Similarly, the prothrombin time (p < 0.05) and the activated partial thromboplastin time (p < 0.05) were significantly more prolonged in rabbits receiving tissue thromboplastin compared with control animals. Also the concentration of blood platelets (p < 0.05), plasma fibrinogen (p < 0.05), antithrombin (p < 0.05), and protein C (p < 0.05) decreased significantly in thromboplastin-treated animals compared with control animals. In all these haemostatic variables there was a common trend that animals infused with tissue thromboplastin in the arcus aorta responded more excessively than animals infused in the right atrium of the heart, and these deviations were statistically significant for fibrinogen (p < 0.05) and prothrombin time (p < 0.05). Similarly, animals infused with tissue thromboplastin in the arcus aorta had an increased number of microthrombi in the lungs and kidneys compared with animals receiving tissue thromboplastin above the right atrium. As the lungs are the first pass organ when you infuse above the right atrium the results from this study suggest that the lungs play a key role in protecting the organism against excessive tissue thromboplastin-induced activation of coagulation.

Risk factors for central nervous system involvement in non-Hodgkins-lymphoma--a multivariate analysis.

A consecutive cohort of patients with NHL was examined to identify the factors predictive of CNS-involvement with Cox's proportional hazards model in a multivariate analysis. Twenty-seven cases of CNS-involvement were found among 498 patients with NHL. Only 3 of 96 patients with low-grade lymphomas had CNS involvement, all occurring after transformation into high-grade lymphoma. In univariate analysis of 402 patients with intermediate or high-grade lymphoma, lymphoblastic histology (including Burkitt's lymphoma), age <35 years, B-symptoms, stage IV disease, testis involvement and bone marrow involvement were found to be statistically significant risk factors. Lymphoblastic histology was found to be strongly correlated to age younger than 35 years. In the multivariate analysis only lymphoblastic histology, stage IV disease and B-symptoms were found to be significantly associated with CNS involvement. It is concluded that CNS prophylaxis should be considered in all patients with lymphoblastic histology and in patients with stage IV B lymphomas other than those of low-grade types.

Translocation (1;16) identified by chromosome painting, and PRimed IN Situ-labeling (PRINS). Report of two cases and review of the cytogenetic literature.

We used the molecular cytogenetic in situ techniques chromosome painting and PRimed IN Situ labeling (PRINS) to elaborate the cytogenetic observations in two cases of the rare aberration der(16)t(1;16), which occurs in a wide variation of hematologic and nonhematologic malignancies [1-3]. Review of the literature showed that, in contrast to the chromosome 1 breakpoint, the breakpoint on chromosome 16 is associated with diagnosis as well as patient age.

[The relation of birth weight and gestational age in a group of infants in Jutland with special reference to diagnosing infants with intrauterine growth retardation]. / Sammenhaengen mellem fødselsvaegt og gestationsalder blandt en gruppe jyske børn med henblik på at diagnosticere intrauterint vaekstretarderede børn.

The relationship between birthweight (BW) and gestational age (GA) in 14,276 Danish children with GA of 35 to 42 completed weeks was illustrated employing 10%, 50% and 90% percentile curves. The curves were sex-specific. The number of light for dates (LFD) children defined by means of the 10% percentile (BW below 10th percentile) was 1,351 compared to 848 children when using the curve currently employed. The latter curve was based upon foreign children and moreover was not sex-specific. It is concluded that sex-specific and updated curves representing the population studied are mandatory in the diagnosis of LFD-children.

[Demonstration of urogenital Chlamydia infection by means of the Chlamydiazyme test]. / Påvisning af urogenital Chlamydia-infektion ved hjaelp af Chlamydiazyme-testen.

Abbott's recently introduced Chlamydiazyme test which is based on the Elisa principal and cell culture technique was employed with the object of demonstrating urogenital infection with Chlamydia trachomatis in 150 persons. Neither of the methods were optimal. The Chlamydiazyme test had a lower sensitivity and specificity than the cell culture method.

[Breech presentation deliveries]. / Fødsel med foster i underkropspraesentation.

A total of 5,519 consecutive single deliveries at term were analysed for maternal and foetal factors associated with breech presentation (UK). The protocol in this department for breech deliveries which permits vaginal delivery of foetuses estimated to be between 2,400 and 3,800 g in cases where the pelvis was considered to be clinically normal, was also evaluated. Breech presentation was found in 173 cases. In 102 of these circumstances were present which permitted trial of vaginal delivery. Seventy-seven were delivered vaginally. An increased frequency of low Apgar scores (less than 8) after one minute was demonstrated among infants delivered vaginally in the breech presentation on comparison with infants delivered by Caesarean section, whereas low scoring after five minutes occurred with the same frequency. One infant died during delivery. Follow-up of infants delivered in breech presentation (mean period of observation two years) showed developmental disturbances in three of the vaginally delivered infants and in five of those delivered abdominally. Two of the infants delivered abdominally had severe cerebral paresis and psychomotor retardation without evidence of intrauterine or severe neonatal asphyxia. On comparison with the population delivered in cephalic presentation (HST), significantly increased frequencies of primiparity and light-for-dates infants were found in the breech presentations. Low Apgar score after one minute was significantly more frequent in breech presentations while low Apgar scoring after five minutes occurred with the same frequency. The perinatal mortality rates in breech and cephalic presentations were 17.3 and 4.7 per 1,000 respectively. Following correction for lethal malformations, the rates were 5.8 and 3.8 per 1,000, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Danazol in the treatment of ureteral endometriosis.

We report 2 cases of ureteral endometriosis in which initial hormonal management, including danazol, failed to reverse or to stop the progressing ureteral stenosis. After reviewing the various methods of treatment presented in the literature we conclude that solid documentation of the effect of danazol against ureteral endometriosis is lacking. To preserve kidney function we recommend resection of the affected portion of the ureter.

First-line diagnosis based on immunological phenotyping in suspected acute leukemia: a prospective study.

In a prospective analysis of the diagnostic value of immunophenotyping in acute leukemias (ALs), all patients admitted to a pediatric and a haematological department suspected of AL were examined consecutively with a selected panel of monoclonal antibodies (Mabs) against leucocyte differentiation antigens during an 8-month period. A total of 189 samples obtained from blood, bone marrow, spinal fluid and lymph nodes in 120 cases were all analysed blindly. The results were correlated with a routine morphological/cytochemical evaluation. Differing results were obtained in seven out of 38 cases in which the immunologically defined diagnosis was acute myeloid leukemia (AML), and in one out of 21 cases with the primary diagnosis acute lymphoid leukemia (ALL). Immunological phenotyping disclosed two cases of hybrid leukemia, one case of biphenotypic and one case of bilineal leukemia. No evidence of malignancy was found in 36 cases, 30 cases of blood and bone marrow and six cases of spinal fluids, in every case in accordance with the pathological examination. These results demonstrate that a first-line immunological evaluation of bone marrow, blood and spinal fluid from patients suspected of AL is highly capable of discriminating between different malignant and nonmalignant haematological diseases and also between various types of leukemias. The immunological methods do, however, require a sufficient amount of material which was a limiting factor in 14 out of 120 examinations, mainly from patients treated with several cycles of cytostatics. It is concluded that immunophenotyping can be used as a first-line diagnostic tool in malignant haematological diseases.

Antibiotic elimination of group-B streptococci in urine in prevention of preterm labour.

The presence of group-B streptococci in the urine of pregnant women seems to be associated with preterm labour. Urine samples from 4122 women at 27-31 weeks' gestation were examined for bacteria. Group-B streptococci were found in the urine of 69 women. In a double-blind, controlled study these patients were given either penicillin (10(6) IU three times daily for 6 days; 37 patients) or placebo (32 patients). The rates of primary rupture of the membranes (11% v 53%; p less than 0.001) and preterm labour (5.4% v 38%; p less than 0.002) were significantly lower in the penicillin group than in the placebo group. These results suggest that treatment and follow-up to prevent recolonisation in pregnant women with group-B streptococci in the urine may reduce the frequency of preterm labour in these patients.

Short stature in Scandinavian women. An obstetrical risk factor.

We have carried out a case-controlled study on relations between short stature (i.e. less than 156 cm tall) and problems with childbirth in Danish women. Data obtained from 182 pregnant, short women (short mothers) were compared with those obtained from a control group of 2116 pregnant women who were between 166 and 175 cm tall (control mothers). The prevalence rate for acute cesarean section was three-fold greater in short mothers than in controls, and the prevalence rate for elective cesarean section was twice as high in short mothers as in controls. Moreover, the prevalence rates of intra-uterine asphyxia, intra-uterine growth retardation and low Apgar scores were higher in babies of short mothers than in those of control mothers, despite the increased level of obstetric intervention in the former group. Since the findings show that short stature in pregnant women is an obstetrical risk factor, we recommend that it should be given attention in order to detect early signs of intra-uterine asphyxia and to apply the best form of active management of labor if necessary.