The effect of crude extract from Radix Dipsaci on bone in mice.

Radix Dipsaci is used in China for the treatment of bone fractures and other bone diseases. The aim of the study was to assess the systemic effect of Radix Dipsaci consumption on bone histomorphology. Twenty eight-week-old male BALB/c mice were divided into two groups. In the control group, ten mice were daily fed with distilled water. In the Radix Dipsaci group, ten mice were daily fed with distilled water mixed with crude Radix Dipsaci extract. The mice were kept for 5 weeks and were then killed. Using microcomputed tomography, 20 microtomographic slices with an increment of 0.25 mm were acquired to cover the proximal end of the left tibia of each mouse. Quantitative morphometry of the bone structure was performed. Histological sections of the region of micro-tomographic slices were prepared. The results showed that the consumption of Radix Dipsaci extract caused a 4.50% increase in bone volume/tissue volume ratio. The bone trabeculae increased by 11.82% in number so that the bone density was increased. To conclude, Radix Dipsaci extract taken orally increased bone density and altered bone histomorphology.

Factors regulating condylar cartilage growth under repeated load application.

Mechanical loading can influence the biological behavior of the bone-associated cells leading to adaptive changes in skeletal mass and architecture. SOX9 and PTHrP genes are known to regulate chondrocyte differentiation and delay maturation, ultimately control the endochondral bone formation. To investigate the effects of repeated mechanical loading on bone, 280 Sprague-Dawley rats were used in this experiment. The animals were randomly allocated into experimental and control groups. Repeated mechanical loading was applied through a bite-jumping device in the experimental group. The experimental animals were sacrificed on 10 different time points together with the matched control. Total RNA was extracted from the mandibular condylar cartilage for PTHrP and SOX9 genes quantification using real-time RTPCR. Results showed that PTHrP expression was increased and reached a peak level on the seventh day after mechanical loading was given. Repeated mechanical loading triggered a significant increase of PTHrP expression leading to another peak increment. The expression of SOX9 was highly correlated with the PTHrP expression, and its pattern of expression was similar to that of PTHrP after repeated mechanical loading. In conclusions, repeated mechanical loading on the condyle triggers the expression of PTHrP and SOX9, which in turn promotes condylar cartilage growth.

Association between insulin resistance and GAD65-autoantibody levels--a pilot study in an adult non-diabetic population.

The aim of this pilot study was to investigate any association between insulin resistance (IR) and serum levels of autoantibodies against the glutamic acid decarboxylase (GAD65Ab) among adult non-diabetic subjects. Based on calculations of IR using the IR homeostasis model in a Swedish adult non-diabetic population (n = 756) participating in the WHO MONICA-study, an insulin sensitive group (n = 54, M/F:27/27) and an insulin resistant group (n = 46 M/F:24/22) were identified. Serum from the subjects were analysed for the presence of GAD65Ab. There was no significant difference in GAD65Ab levels between the groups. However, there was a correlation between IR and serum GAD65Ab within the insulin sensitive group (Spearman rho 0.4, p < 0.01). Our observation could indicate that IR could serve as an initiator or a progression factor in the autoimmune process in subjects predisposed to autoimmunity. This finding will be further investigated in a larger study including subjects with a continuum of IR.

High GAD65 autoantibody levels in nondiabetic adults are associated with HLA but not with CTLA-4 or INS VNTR.

OBJECTIVES: To explore the relationship between genetic background and antibody levels in a nondiabetic population. We evaluated if high levels of autoantibodies against the 65 kDa isoform of glutamic acid decarboxylase (GAD65Ab), were associated with high-risk genes, i.e. HLA, CTLA-4 and INS VNTR genes. DESIGN AND SUBJECTS: Seventy-five (M/F 39/36) subjects exceeding the 95th percentile of GAD65 autoantibody index and 75 age and sex matched subjects below the 95th percentile, randomly selected amongst participants in the Västerbotten Intervention Programme. METHODS: The GAD65 Ab were measured in a radioligand-binding assay. HLA class II typing was performed by an oligoblot hybridization method. CTLA-4 repeat length was analysed and divided into short forms and long forms. Class I and class III alleles of INS VNTR were detected. Differences in distribution were tested by Pearson chi-square with Yates correction. Odds ratios (OR) were used to compare groups calculated with Cochran's and Mantel-Haenszel statistics. RESULTS: The DQB1*0201-DQA1*0501-DRB1*03 haplotype was increased in subjects with high GAD65Ab levels (P = 0.04). This increase seemed to be explained by a difference in haplotype frequencies amongst men (P = 0.01). Calculating OR showed a significant association between the DQB1*0201-DQA1*0501-DRB1*03 haplotype and elevated levels of GAD65Ab in all subjects (OR 2.2, 95% CI 1.02-4.9) as well as in men (OR 4.6, 95% CI 1.3-15.9). There was no association between high levels of GAD65Ab and either INS VNTR or CTLA-4 polymorphisms. CONCLUSION: Our study suggests that adult males with the DQB1*0201-DQA1*0501-DRB1*03 haplotype tend to develop high GAD65Ab titres. As none of these subjects have developed diabetes these data suggest that HLA may be important in GAD65Ab formation but that additional factors are required for the progression to overt type 1 diabetes.

Streptozotocin induced diabetes in minipig: a case report of a possible model for type 1 diabetes?

Studies on the pathogenic process in type 1 diabetes are often performed in animal models. Low-dose administration of streptozotocin has been used to induce diabetes with pathological alterations similar to human type 1 diabetes in the animals. Rodent models are frequently used but there is a need of developing new models including larger animals. In this study we wanted to investigate to what extent a minipig was sensitive to low-dose streptozotocin for induction of diabetes with features of human Type I diabetes. A female Göttingen minipig received two low-doses (40 mg/kg) of streptozotocin with an 11-day interval. Serum was analysed for the presence of the enzyme glutamic acid decarboxylase, isoform 65, (GAD65) and autoantibodies against glutamic acid decarboxylase, isoform 65 (GAD65A), isoform 67 (GAD67A), insulinoma antigen 2 (IA-2) and insulin (IAA). Pancreas tissue was fixated in formaldehyde and was sent for pathoanatomical examination. The minipig became hyperglycaemic after the second injection of streptozotocin. The pathoanatomical examination showed atrophy of the beta-cell population, depletion of insulin with preserved glucagon content. There was no sign of insulitis. Both GAD65 and GAD65A were detected while GAD67A and IAA were absent. It is concluded that chronic diabetes developed after low-dose streptozotocin injection in a female minipig with the characteristics of the end stage of type 1 diabetes. This pilot study suggests that minipigs show promise as a model to induce diabetes by injections of low-dose streptozotocin.

Bone induction in clinical orthodontics: a review.

The major limitations of autogenous grafting are inadequate supply and surgical morbidity, including donor site pain, paresthesia, and infection. Graft resorption can also pose a severe problem. Bone induction is therefore needed to assist in fracture healing and to fill osseous defects. This article reviewed the current development of bone induction in relation to clinical orthodontics.

Osteogenesis in the glenoid fossa in response to mandibular advancement.

The purpose of this study was to identify the temporal sequence of cellular changes in the glenoid fossa and to quantify the amount of bone formation in response to mandibular advancement. One hundred 35-day-old female Sprague-Dawley rats were randomly divided into 5 experimental groups (15 rats each) and 5 control groups (5 rats each). In the experimental groups, functional appliances were used to create continuous forward mandibular advancement. The rats were killed after 3, 7, 14, 21, and 30 days. Sections were cut through the glenoid fossa in the parasagittal plane and stained with periodic acid and Schiff's reagent for evaluation of bone formation and with hematoxylin and eosin for observation of cellular response. The results showed that, in the control rats, bone formation was initially higher in the posterior and middle regions than in the anterior region then decreased over time in all regions. In the experimental group, bone formation significantly increased from day 7 to day 30 compared with control rats. Day 21 marked the highest levels of bone formation in the middle (+184%) and posterior regions (+300%). Mandibular protrusion resulted in the osteoprogenitor cells being oriented in the direction of the pull of the posterior fibers of the disc and also resulted in a considerable increase in bone formation in the glenoid fossa.

Prediction of diabetes with body mass index, oral glucose tolerance test and islet cell autoantibodies in a regional population.

OBJECTIVE: Our aim was to test the hypothesis that a combination of markers for Type 1 diabetes (glutamate decarboxylase and IA-2 autoantibodies) and for Type 2 diabetes [oral glucose tolerance test (OGTT) and body mass index (BMI)], would predict clinical diabetes in a regional population. DESIGN: A population-based follow-up cohort study. SETTING: Participants visited the primary health care centre in Lycksele, Sweden in 1988-92. PARTICIPANTS: A cohort of 2278 subjects (M/F 1149/1129) who were studied at follow-up in 1998. At base line there were 2314 subjects (M/F 1167/1147) who participated in the Västerbotten Intervention Program on their birthday when turning either 30, 40, 50 or 60 years of age. Main outcome measurements. A clinically diagnosed diabetes at follow-up when the medical records were reviewed for diagnosis of diabetes. At base line, the participants were subjected to a standard OGTT and their BMI determined along with the autoantibodies. RESULTS: At follow-up, 42/2278 (1.8%, 95% CI 1.2-2.3) (M/F 23/19) had developed diabetes: 41 subjects were clinically classified with Type 2 and one with Type 1 diabetes. There was no significant relation between autoantibody levels at base line and diabetes at follow-up. Stepwise multiple logistic regression showed that the odds ratio for developing diabetes was 10.8 (95% CI 6.3-18.9) in subjects in the fourth quartile of BMI (BMI > 27) compared with 7.8 (95% CI 4.8-12.6) in the fourth quartile of 2-h plasma glucose (>7.5 mmol L(-1)) and 7.2 (95% CI 4.8-11.4) in the fourth quartile of the fasting plasma glucose (>5.6 mmol L(-1)). CONCLUSION: Islet cell autoantibodies did not predict diabetes at follow-up. BMI measured at base line was as effective as 2-h plasma glucose and fasting plasma glucose to predict diabetes in this adult population.

Growth hormone replacement therapy improves body composition and increases bone metabolism in elderly patients with pituitary disease.

Although a specific GH deficiency (GHD) syndrome in the adult and the response to GH replacement therapy are well recognized, there are few data available on the effect of GH replacement therapy in elderly GH-deficient patients. We studied the effect of GH therapy on body composition and bone mineral density measured by dual energy x-ray absorptiometry, markers for bone metabolism, insulin-like growth factors (IGFs), and IGF-binding proteins (IGFBPs) in 31 patients (6 women and 25 men; aged 60-79 yr; mean, 68 yr) with multiple pituitary hormone deficiencies. The GH response to arginine or insulin was below 3 microg/L (9 mU/L) in all subjects. They were randomized to GH (Humatrope, Eli Lilly & Co.) or placebo for 6 months, followed by 12 months of open treatment. The dose was 0.05 IU/kg x week for 1 month, and after that it was 0.1 IU/kg x week divided into daily sc injections (0.75-1.25 IU/day). There were no changes in any of the measured variables during placebo treatment. GH treatment normalized serum IGF-I in a majority of the patients and increased IGFBP-3 and -5 as well as IGFBP-4 and IGF-II to values within normal range. Lean body mass was increased, and the increase at 6 and 12 months correlated with the increase in IGF-I (r = 0.46; P = 0.010 and r = 0.54, respectively; P = 0.003). GH treatment caused a modest, but highly significant, reduction of total body fat. Mean bone mineral density was not different from that in healthy subjects of the same age and did not change during the observation period. Markers for bone formation (bone-specific alkaline phosphatase activity, osteocalcin, and procollagen I carboxyl-terminal peptide in serum) increased within the normal range, and levels were sustained throughout the study. The bone resorption marker (pyridinoline in urine) was significantly elevated for 12 months. Side-effects were mild, mostly attributed to fluid retention. In two patients with normal glucose tolerance at the start of the study, pathological glucose tolerance occurred in one patient and was impaired in one. In conclusion, elderly patients with GHD respond to replacement therapy in a similar manner as younger subjects, with an improvement in body composition and an increase in markers for bone metabolism. Side-effects are few, and elderly GHD patients can be offered treatment. As long-term risks are unknown, GH doses should be titrated to keep IGF-I within the age-related physiological range.

Magnesium therapy in type 1 diabetes. A double blind study concerning the effects on kidney function and serum lipid levels.

To test the hypothesis that magnesium depletion might be of importance for the development of vascular complications in diabetes mellitus we performed a randomized, double-blind, placebo-controlled study during 12 months with 20-30 mmol/day of oral magnesium hydroxide in 28 type 1 diabetic patients. Urinary albumin excretion, Cr-EDTA-clearance and certain blood cardiovascular risk factors were measured. At the end of the study there were no significant differences of these parameters between the two groups, except that serum triglyceride values increased in three magnesium treated patients who either showed an increase in blood glycosylated hemoglobin values or body weight during the study.

Glutamate decarboxylase (GAD65) and tyrosine phosphatase-like protein (IA-2) autoantibodies index in a regional population is related to glucose intolerance and body mass index.

AIMS/HYPOTHESIS: Our aims were to investigate the concentrations and prevalence of autoantibodies against the Mr 65.000 isoform of glutamate decarboxylase (GAD65) and the tyrosine phosphatase-like protein (IA-2) in adults and to test the hypothesis that GAD65 and IA-2 autoantibodies in a regional population are related to abnormal oral glucose tolerance. METHODS: We analysed serum from 2157 Swedish subjects aged either 30, 40, 50 or 60 years old who, in 1988-1992, participated in the Västerbotten County Health Project and were subjected to the World Health Organisation (WHO) standard oral glucose tolerance test at entry into the study. RESULTS: We found 23 of 2157 (1.1%) and 17 of 2152 (0.8%) subjects exceeded the 99th centile of GAD65 autoantibody index and IA-2 autoantibody index, respectively. In 18 subjects with diabetic oral glucose tolerance test, GAD65 autoantibody concentrations were higher than in those with normal oral glucose tolerance test (p = 0.02). Subjects with IGT (n = 185) [corrected] and diabetes (n = 18), i.e. abnormal OGTT (n = 203) [corrected], had higher GAD65Ab [corrected] index compared with those with normal OGTT (p = 0.026) [corrected]. A stepwise multiple logistic regression test showed that the odds ratios for subjects in the highest BMI group to exceed the 95th or 99th GAD65 autoantibody centile were 3.6 (CI 1.4-8.9) and 17.6 (CI 2.6-121.6), respectively. CONCLUSION/INTERPRETATION: GAD65 and IA-2 autoantibodies, are associated with impaired or diabetic glucose tolerance in an adult regional population. This observation together with the association between GAD65 autoantibody concentrations and body mass index indicate a possible relation between islet autoimmunity and beta-cell function abnormalities with obesity and insulin resistance.

Aberrant V(H) gene utilization in patients with established insulin dependent diabetes mellitus.

We have compared the B-lymphocyte repertoire in seven IDDM patients with 12 healthy controls by examining the variable heavy (V(H)) gene expression. The V(H) gene representation in the pool of pokeweed mitogen (PWM) stimulated, immunocompetent B cells and in the pool of naturally activated plasma cells (actual repertoire) was analysed by RNA-RNA in situ hybridization. Differences between IDDM patients and normal controls in the relative expression of several V(H) gene families were observed. In IDDM patients, the V(H)3 was significantly underrepresented in the PWM stimulated repertoire. In the actual B cell repertoire the V(H)5 clones were underrepresented among diabetic patients. Moreover, the altered distribution of V(H) gene usage between the PWM stimulated repertoire and the actual repertoire observed in normal controls was found to be less pronounced in the IDDM patients. This observation suggests a defect in the V-gene directed cellular selection occurring between resting, immunocompetent B cells and naturally activated plasma cells. The possible implication of the observed aberrations in the B cell selection process for the pathogenesis of autoimmunity is discussed.

A low dose ACTH test to assess the function of the hypothalamic-pituitary-adrenal axis.

OBJECTIVE: The insulin tolerance test (ITT) has long been used to assess the hypothalamic-pituitary-adrenal axis, but may be hazardous. The standard synthetic ACTH (Synacthen) test has been advocated as a substitute but is sometimes insensitive. In this study the ITT has been compared to a low dose ACTH stimulation test (1 microg) and the standard ACTH stimulation test (250 microg). SUBJECTS: Twenty-seven subjects were studied, 24 with verified or suspected hypothalamic-pituitary disorders and three on long-term glucocorticoid therapy. DESIGN: Insulin tolerance, low dose ACTH and standard ACTH tests were performed in all patients. The ITT was performed less than 48 hours after the ACTH tests. Synacthen was administered as an intravenous bolus. MEASUREMENTS: Serum cortisol values were determined by radioimmunoassay. The peak cortisol value during ITT was compared to the cortisol levels during the ACTH tests. RESULTS: There was a highly significant correlation between peak cortisol values during ITT and cortisol levels after 20-60 minutes in the low dose ACTH test (r(s) = 0.91-0.93; P < 0.0001) and after 30 and 60 minutes in the standard ACTH test (r(s) = 0.85 and 0.89 respectively; P < 0.0001). Four patients showed discrepancies between the three tests. CONCLUSIONS: The 1-microg ACTH test follows the ITT more closely and may be more sensitive than the standard ACTH test in detecting more subtle insufficiency of the hypothalamic-pituitary-adrenal axis. The standard ACTH test and the insulin tolerance test may thus be replaced by the 1-microg ACTH test in screening for secondary cortisol insufficiency. We recommend that serum cortisol is measured before and 30 and 40 minutes after the ACTH injection.

Quality of life in adults with growth hormone (GH) deficiency: response to treatment with recombinant human GH in a placebo-controlled 21-month trial.

We examined the effect of GH supplementation on the psychological capacity and sense of well-being in 36 patients with adult-onset GH deficiency (GHD). Recombinant human GH was given in a 21-month cross-over, double blind trial, and quality of life was assessed by using three self-rating questionnaires: the Hopkins Symptom Check List (HSCL), the Nottingham Health Profile (NHP), and the Psychological General Well-Being index. In addition, at the final examination the spouses completed a short questionnaire concerning their partner. Before treatment, the patients had lowered quality of life as determined by the HSCL and NHP inventories, and a correlation between the duration of GHD and the reported symptoms was observed. Upon treatment, the HSCL score was lower (better) after placebo administration (mean +/- SD, 84 +/- 21.3) than at baseline (89 +/- 18.9; P = NS) and fell to 80.2 +/- 18.5 (P < 0.001) when active drug was given. The subscales regarding anxiety, fearfulness, and cognition were the most sensitive. It was apparent that the effect determined after GH therapy in part was due to a placebo effect. With NHP, the dimensions of energy and emotions responded most to treatment. Further, the spouses observed their partners to be improved in several aspects of mood and behavior (P < 0.05 to P < 0.0001) when active drug was given. The data thus demonstrate that GH, which is known to have multiple somatic effects, produces an improvement in the quality of life of adults with GHD.

Soft tissue changes following maxillary osteotomies in cleft lip and palate and non-cleft patients.

A retrospective study of 25 patients with unilateral cleft lip and palate and 25 patients with hypoplastic maxillae without a cleft was carried out to evaluate the effects of maxillary osteotomies at the Le Fort I level on the lip and nose profile. The pre-surgical cephalometric tracing was superimposed twice on the post-surgical cephalogram, on the cranial structures and the anterior maxillary structures, for landmark movement measurement. Results showed statistically significant correlations between soft and hard tissue movement in the cleft group. On average, the ratios of horizontal nasal tip, nasal base and lip movement to underlying hard tissue movement were approximately one fourth, one half and two thirds respectively. The ratio of vertical lip to incisor movement was about one half. The correlations were less significant in the non-cleft group, only the upper lip movement showed statistically significant correlation with hard tissue movement, with a ratio of one half horizontally and one third vertically. It was concluded that in maxillary osteotomy, the cleft group showed a higher soft tissue to hard tissue movement ratio. The correlation between soft and hard tissue movements were more statistically significant in the cleft group than in the non-cleft group. However, though statistically significant, the level of correlation was not strong on an individual basis except in the horizontal lip response of the cleft group. Individual variation was wide and clinical judgement needs to be considered accordingly.

Prediction of caries in 1 1/2-year-old children.

The aim was to test the predictive ability of defined levels of dietary and oral hygiene habits and also mutans streptococci and lactobacilli in saliva, separately and in combinations, in 1 1/2-year-old children. The population consisted of 181 children, who were investigated with respect to the above factors together with general health, use of fluoride in tablets and/or toothpaste, gingival condition and caries prevalence. Initially, 99% of the subjects were caries-free as against 72% at the age of 3 years, with a mean of 0.8 ds. Using data for sensitivity and specificity the predictive values for positive (PV+) and negative (PV-) tests were calculated for different levels and combinations of variables. A dividing line between high and low caries risk that combined high sensitivity and high specificity could not be established for any variable or combination of variables. High sensitivity was noted with diet as the predictor, high specificity with oral hygiene or occurrence of mutans streptococci. Lactobacilli were excluded as they were found in only 6 children. A two-step computation with mutans streptococci as second predictor improved the ability to single out children with caries at the age of 3 years. Subgrouping and analyses of the material as regards use of fluoride did not influence the results. To sum up, prediction at the age of 1 1/2 years, in a population with a low caries prevalence, was not successful with the variables used in this study.

The prognostic value of admission blood pressure in patients with acute stroke.

BACKGROUND AND PURPOSE: Patients with acute stroke are often found to have high blood pressures at hospital admission. Previous studies have shown variable results regarding the prognostic value of high blood pressure in acute stroke. The aim of this study was to investigate the prognostic value of admission blood pressure in a population-based sample of patients with acute stroke. METHODS: Eighty-five patients with intracerebral hemorrhage and 831 with ischemic disease were included in the study. The relations between admission blood pressure and 30-day mortality were studied by logistic regression analyses. RESULTS: High blood pressure in patients with impaired consciousness on hospital admission was significantly related to 30-day mortality in patients with intracerebral hemorrhage (P = .037) and in patients with ischemic disease (P < .0001). In patients without impaired consciousness, high blood pressure at time of admission was not related to increased mortality at 30 days. CONCLUSIONS: High admission blood pressure in alert stroke patients was not related to increased mortality. Stroke patients with impaired consciousness showed higher mortality rates with increasing blood pressure. However, this does not provide a basis for recommending antihypertensive therapy for such patients.