The Tubifex tubifex assay for the determination of acute toxicity

An express (3-minute) test for acute toxicity determination by using the oligochaete annelid, Tubifex tubifex, is described. The EC50(Tubifex tubifex) [EC50(Tt)] for movement inhibition was calculated by using a concentration-response dependence. The reproducibility of the test was checked over several years and by several workers. Its applicability is limited to compounds which are soluble in water. The calculated EC50(Tt) indices correlate with LC50 values determined by using the fish, Pimephales promelas (96-hour assay), and with ICG50 values determined by using the ciliate, Tetrahymena pyriformis (48-hour assay) with high statistical significance (r = 0.822, n = 35, and r = 0.927, n = 80, respectively). The correlation between the EC50(Tt) indices and rat oral LD50 values (48-hour assay) was r = 0.519 (n = 67). The correlation within organic compounds was closer (r = 0.635, n = 60) than with the heterogeneous series of chemicals. A similar trend was noticed for the correlation with mouse oral LD50 values (r = 0.479, n = 56) with the heterogeneous series of chemicals, as compared that with the series without inorganic salts (r = 0.605, n = 42), and similarly with mouse intraperitoneal LD50 values, where r = 0.543 (n = 50) with the heterogeneous series of chemicals and r = 0.893 (n = 33) with the series of organic chemicals.     

Ethnicity and weight status affect the accuracy of proxy indices of insulin sensitivity

This study tested the hypotheses that correlations between direct measures of insulin sensitivity and proxy indices of insulin sensitivity derived from fasting values, (i) would not be affected by ethnicity, and (ii) would be stronger in overweight vs. weight-reduced states. We further hypothesized that associations between proxy indices and fat distribution would be similar to those between directly measured insulin sensitivity and fat distribution. Testing was performed in weight-stable conditions in 59 African-American (AA) and 62 white-American (WA) overweight, premenopausal women before and after a weight loss intervention. Subjects were retested 1 year following weight loss. Proxy indices were correlated against the insulin sensitivity index S(I) determined via minimal modeling. Fat distribution was assessed using computed tomography. Correlations between Si and proxy indices were consistently stronger among overweight women (r = 0.44-0.52) vs. weight-reduced women (r = 0.18-0.32), and among AA (r = 0.49-0.56, baseline; 0.24-0.36, weight-reduced) vs. WA (r = 0.38-0.46, baseline; 0.19-0.31, weight-reduced). Among subjects who regained >3 kg after 1 year, correlations between S(I) and proxy indices were similar to those observed at baseline, whereas correlations were weak among women who maintained their reduced body weight. S(I) and all proxy indices were similarly correlated with intra-abdominal adipose tissue (IAAT) at baseline, but not after weight loss. In conclusion, correlations between S(I) and proxy indices were affected by both ethnicity and weight status. If proxy indices are used in multiethnic populations, or in populations including both lean and overweight/obese subjects, data should be interpreted with caution.     

Predictability of heterozygosity scores and polymorphism information content values for rat genetic markers

Construction of a genetic linkage map of the laboratory rat, Rattus norvegicus, establishes the rat as a genetic model. Allele sizes were reported for 432 simple sequence length polymorphisms (SSLPs) genotyped in 12 different substrains belonging to nine different inbred strains of rats. However, these nine strains represent only a fraction of the more than 140 inbred strains available. If allele sizes are not known, alternative indices of markers' polymorphism content can be used, such as heterozygosity (H) and polymorphism information content (PIC). Here, we have determined heterozygosity scores and PIC values for all markers of the rat genetic linkage map, and we evaluate the predictability of the heterozygosity and the PIC values. Correlation analysis between the nine inbred strains reported for the rat map and ten test strains yielded r=0.42 and r=0.44 for heterozygosity and PIC values, respectively. While the correlation of the indices between the two groups of animals is low, these indices do provide a means of predicting whether a genetic marker will be informative in strains where allele sizes are not known.     

Reliability and validity of a new repeated agility test as a measure of anaerobic and explosive power

The aim of this study was to evaluate the reliability and validity of a repeated modified agility test (RMAT) to assess anaerobic power and explosiveness. Twenty-seven subjects (age: 20.2 ± 0.9 years, body mass: 66.1 ± 6.0 kg, height: 176 ± 6 cm, and body fat: 11.4 ± 2.6%) participated in this study. After familiarization, subjects completed the RMAT consisting of 10 × 20-m maximal running performances (moving in forward, lateral, and backward) with ~25-second recovery between each run. Ten subjects performed the RMAT twice separated by at least 48 hours to evaluate relative and absolute reliability and usefulness of the test. The criterion validity of the RMAT was determined by examining the relationship between RMAT indices and the Wingate anaerobic test (WAT) performances and both vertical and horizontal jumps. Reliability of the total time (TT) and peak time (PT) of the RMAT was very good, with intraclass correlation coefficient > 0.90 and SEM < 5% and low bias. The usefulness of TT and PT of the RMAT was rated as "good" and "OK," respectively. The TT of the RMAT had significant correlations with the WAT (peak power: r = -0.44; mean power: r = -0.72), vertical jumps (squat jump: r = -0.50; countermovement jump: r = -0.61; drop jump (DJ): r = -0.55; DJ with dominant leg: r = -0.72; DJ with nondominant leg: r = -0.53) and 5 jump test (r = -0.56). These findings suggest that the RMAT is a reliable and valid test for assessing anaerobic power and explosiveness in multisprint sport athletes. Consequently, the RMAT is an easily applied, inexpensive field test and can provide coaches and strength and conditioning professionals with relevant information concerning the choice and the efficacy of training programs.     

Reaction of Cardiac Activity of Senior Pupils of Schools Providing Differentiated Education upon Examination Stress

Effects of examination stress on psychological characteristics and indices of the autonomic nervous system were studied in pupils of the eighth and tenth grades of high schools providing differential (test group, Neuron gymnasium) and general education (control group). General health state and levels of activity and mood (feeling, activity, mood; FAM) and neuroticism (according to Eysenck) were determined in both groups. Main hemodynamic indices (heart rate and blood pressure) were measured, and mathematical analysis of the heart rate by was performed according to Baevskii. Comparative analysis of psychophysiological tests and heart rate changes in the control and test groups points to a high tension in the cardiac rhythm regulation mechanisms and strong anxiety in schoolchildren in the differential education school. The character and depth of preexamination stress depended on the type of autonomic control of cardiac activity. Schoolchildren with sympathicotonic cardiac rhythm regulation showed the strongest emotional tension. Schoolchildren with the normotonic type of regulation were the most calm.     

Assessment of binding indices and physiological responsiveness of the 5-HT2 receptor on human platelets

This is the first report of parallel studies of binding indices and physiological responsiveness of the "Serotonin-two" (5-HT2) receptor on the human platelet membrane. Binding indices were measured by a microassay employing [125I]ILSD as radioligand and ketanserin to define specific binding. A single receptor population was found, characterized by a KD of 1.69 +/- 0.45 nM and Bmax of 14.5 +/- 6.0 pmol/g protein in healthy subjects. Functional responsiveness of the platelet 5-HT2 receptor complex was assessed by measurement of the extent to which serotonin (10uM) augmented platelet aggregation induced by threshold concentrations of adenosine diphosphate (ADP). A statistically significant positive correlation was found between the number of platelet 5-HT2 receptor sites (Bmax) and the magnitude of the serotonin-amplified aggregation response (r = .70, n = 38, p less than 0.001). Assessment of binding indices and physiological responsiveness of the platelet 5-HT2 receptor complex should facilitate study of age, hormonal, disease, and drug effects on 5-HT2 receptor function in human subjects.     

Mineralization and wear of mandibular first molars in red deer (Cervus elaphus) of known age

The degree of dental wear was measured in 41 mandibular first molars from red deer of known age (range 2–18 years). The calculated wear indices were positively correlated with age ( r = 0.780, P < 0.001). In the same sample, significant ( P < 0.001) differences in hardness (determined by microhardness testing) were observed between enamel and coronal dentine as well as between outer and inner enamel and dentine, respectively. In animals belonging to the same age group (three-year-olds, n = 11), a negative correlation ( r = -0.830, P = 0.0016) was found between wear index and maximum mean enamel hardness. In a sample of five specimens from this age group, a negative correlation ( r = -0.918, P = 0.028) between maximum calcium concentration of enamel (measured by electron microprobing) and wear index was also recorded. We concluded that the intensity of enamel mineralization had a decisive effect on the degree of dental wear. Measuring of enamel hardness and subsequent modification of wear scores or indices is therefore recommended for future studies on the relationship between age and the degree of dental attrition in deer.     

The correlation between the frequency of sister chromatid exchanges and the kinetics of human lymphocyte proliferation: the dependence on the sex of the donor

Sister chromatid exchanges (SCE) and average generation time (AGT) were studied in lymphocytes from 35 donors (23 females and 12 males). A higher SCE frequency was found in lymphocytes from females than from males. Smoking increased SCE frequency in lymphocytes of males, but not of females. No differences in AGTs between males and females were found. Partial correlation coefficients between SCE frequency, AGT values, donor's age and smoking were determined. A statistically significant correlation (r = 0.650, P less than 0.01) between SCE frequency and AGT was found in lymphocytes obtained from females. In lymphocytes from males statistically significant partial correlation coefficients were detected between SCE frequency and AGT (r = -0.696, P less than 0.05), SCE frequency and donor's age (r = 0.770, P less than 0.01), SCE frequency and smoking intensity (r = 0.697, P less than 0.01), AGT value and donor's age (r = 0.882, P less than 0.01), and AGT value and smoking (r = 0.634, P less than 0.05). Thus, considerable differences in number of indices between males and females exist. The present observations together with other studies (D'Souza et al., 1988) suggest that considerations for population monitoring using cytogenetic techniques (ICPEMC Publication No 14) may be supplemented with the recommendation to use (whenever it possible) only males as donors in population studies.     

Echo-Doppler ultrasonographic assessment of resistance and velocity of blood flow in the ductus venosus throughout gestation in fetal lambs

It has been postulated that during fetal hypoxia, the blood flow shunted through the ductus venosus increases and may account for upto 70% of the total umbilical flow. The objectives of the present study were to use ultrasonography to determine the velocities and waveform indices of blood flow in the ductus venosus in the fetal lamb. The ductus venosus of 15 lamb fetuses was evaluated weekly from Days 45 to 143 of gestation (Day 0=day of AI). The Doppler indices measured were: S/D, the systole/diastole ratio; RI, the resistance index; and PI, the pulsatility index. The velocity waveforms studied were: SV, the peak velocity during ventricular systole; D, the peak velocity during ventricular diastole; aV, the lowest forward velocity during atrial contraction; and TAMV, the time-averaged maximum velocity. Doppler indices from Days 52 to 143 were highly correlated: S/D versus PI (r=0.96, P<0.0001), and versus RI (r=0.93, P<0.0001); and PI versus RI (r=0.97, P<0.0001). Velocity indices were also positively correlated: velocity SV versus D (r=0.87, P<0.0001), versus aV (r=0.79, P<0.05), and versus TAMV (r=0.98, P<0.0001); D versus aV (r=0.88, P<0.05), and versus TAMV (r=0.87, P<0.05); and aV versus TAMV (r=0.82, P<0.05). Doppler indices were negatively correlated with SV (r=-0.22, P<0.03); D (r=-0.37, P<0.0001); TAMV (r=-0.32, P<0.05) and with aV (r=-0.67, P<0.05). The mean value of each Doppler index decreased 40% from Days 52 to 73 (e.g., PI from 0.82+/-0.08 to 0.51+/-0.10; P<0.05), with no significant changes thereafter. Mean (+/-S.E.M.) values of velocity indices SV, D, aV, and TAMV rose twofold from Days 60 to 115 of gestation (e.g., SV from 54.4+/-8.8cm/s to 104.9+/-19.7 and aV from 24.8+/-6.9 to 54.9+/-5.9; P<0.05). In conclusion, Doppler ultrasonography facilitated assessment of the blood flow pattern in the ductus venosus in lamb fetuses between Days 52 and 143.     

Study of insulin-like growth factor I in human obesity.

In obesity there is a decrease in basal and stimulated GH secretion. IGF-I, which has negative feedback effects on GH secretion, could be the initial mediator of such alterations. We studied IGF-I levels in obese subjects and their relationship to the obesity level and GH secretion. We determined plasma IGF-I, basal and stimulated GH in 30 normal and 30 obese women and related these variables to obesity indices (body mass index, BMI, and % overweight). Baseline plasma GH values were 1.2 +/- 0.3 and 2.3 +/- 0.6 micrograms/l in obese subjects and controls, respectively (NS). Mean peak GH secretion after stimuli were 11.2 +/- 1.4 and 34.4 +/- 5.6 micrograms/l in obese subjects and controls, respectively (p less than 0.001). Plasma IGF-I were 1.0 +/- 0.1 U/ml and 0.7 +/- 0.1 U/l in obese subjects and controls, respectively (NS). There was a significant negative correlation between plasma IGF-I and age (r = -0.55, p less than 0.001) and a significant negative correlation between mean peak GH secretion and weight (r = -0.60, p less than 0.001), BMI (r = -0.64, p less than 0.001) and percentage of ideal body weight (r = -0.67, p less than 0.001). We did not find any correlation between IGF-I and indices of overweight. These data suggest that the reduced GH secretion found in obesity is not related to a negative feedback inhibition by elevated levels of IGF-I and that adiposity is not associated with a decline in IGF-I levels. We confirm the existence of a negative correlation between GH secretion and obesity indices.     

Quantifying the improvement of surrogate indices of hepatic insulin resistance using complex measurement techniques

We evaluated the ability of simple and complex surrogate-indices to identify individuals from an overweight/obese cohort with hepatic insulin-resistance (HEP-IR). Five indices, one previously defined and four newly generated through step-wise linear regression, were created against a single-cohort sample of 77 extensively characterised participants with the metabolic syndrome (age 55.6 ± 1.0 years, BMI 31.5 ± 0.4 kg/m(2); 30 males). HEP-IR was defined by measuring endogenous-glucose-production (EGP) with [6-6(2)H(2)] glucose during fasting and euglycemic-hyperinsulinemic clamps and expressed as EGP*fasting plasma insulin. Complex measures were incorporated into the model, including various non-standard biomarkers and the measurement of body-fat distribution and liver-fat, to further improve the predictive capability of the index. Validation was performed against a data set of the same subjects after an isoenergetic dietary intervention (4 arms, diets varying in protein and fiber content versus control). All five indices produced comparable prediction of HEP-IR, explaining 39-56% of the variance, depending on regression variable combination. The validation of the regression equations showed little variation between the different proposed indices (r(2) = 27-32%) on a matched dataset. New complex indices encompassing advanced measurement techniques offered an improved correlation (r = 0.75, P<0.001). However, when validated against the alternative dataset all indices performed comparably with the standard homeostasis model assessment for insulin resistance (HOMA-IR) (r = 0.54, P<0.001). Thus, simple estimates of HEP-IR performed comparable to more complex indices and could be an efficient and cost effective approach in large epidemiological investigations.     

Limited predictive ability of surrogate indices of insulin sensitivity/resistance in Asian-Indian men

Insulin resistance is highly prevalent in Asian Indians and contributes to worldwide public health problems, including diabetes and related disorders. Surrogate measurements of insulin sensitivity/resistance are used frequently to study Asian Indians, but these are not formally validated in this population. In this study, we compared the ability of simple surrogate indices to accurately predict insulin sensitivity as determined by the reference glucose clamp method. In this cross-sectional study of Asian-Indian men (n = 70), we used a calibration model to assess the ability of simple surrogate indices for insulin sensitivity [quantitative insulin sensitivity check index (QUICKI), homeostasis model assessment (HOMA2-IR), fasting insulin-to-glucose ratio (FIGR), and fasting insulin (FI)] to predict an insulin sensitivity index derived from the reference glucose clamp method (SI(Clamp)). Predictive accuracy was assessed by both root mean squared error (RMSE) of prediction as well as leave-one-out cross-validation-type RMSE of prediction (CVPE). QUICKI, FIGR, and FI, but not HOMA2-IR, had modest linear correlations with SI(Clamp) (QUICKI: r = 0.36; FIGR: r = -0.36; FI: r = -0.27; P < 0.05). No significant differences were noted among CVPE or RMSE from any of the surrogate indices when compared with QUICKI. Surrogate measurements of insulin sensitivity/resistance such as QUICKI, FIGR, and FI are easily obtainable in large clinical studies, but these may only be useful as secondary outcome measurements in assessing insulin sensitivity/resistance in clinical studies of Asian Indians.     

Regulation of cell division in the subapical shoot meristem of dwarf watermelon seedlings by gibberellic acid and polyethylene glycol 4000

The stimulatory effects of gibberellic acid (GA₃) and the inhibitory effects of polyethylene glycol 4000 (PEG) on hypocotyl elongation and cell cycle kinetics in subapical pith cells of dwarf watermelon seedlings (Citrullus lanatus [Thunb.] Matsu and Nakai) were investigated. Mitotic indices (MI) were determined from direct counts of pith cells stained by a modified Feulgen technique. Labeling indices (LI) were determined from direct counts of labeled pith cells sampled 1.5 h after apical applications of³H-thymidine. Root application of 0.32 mM GA₃ at 96, 120, or 144 h after sowing resulted in significant increases in both mitotic and labeling indices within 4.5 to 7.5 h following treatment. A single mitotic peak at 13.5 h occurred in all three treatment periods. Labeling peaks were often less defined than mitotic peaks; however, a relatively high proportion of labeled nuclei were usually observed between 7.5 and 9 h after GA₃ treatment and at 16.5 h, the latter period coinciding with progression of cells into S phase from the peak period of mitosis. The results suggest that GA₃ increases the proportion of rapidly dividing cells in the subapical meristem by increasing the probability that slowly cycling or nonproliferative cells in both 2C and 4C DNA states will enter the proliferative pool. The addition of PEG (200 g/l, = 1.5 mPA) to the rooting medium of dwarf watermelon seedlings inhibited hypocotyl elongation and reduced both mitotic and labeling indices simultaneously within 4.5 h after treatment. Within 24–28 h after PEG treatment, mitotic and labeling indices approached 0. Seedlings transferred from PEG to either water or GA₃ exhibited rapid recovery of cell division and hypocotyl elongation. Mitotic and labeling indices increased within 4.5–7.5 h into the recovery period in either water or GA₃ and reached control values within 10.5 h. GA₃ hastened the recovery from PEG-induced stress. It is concluded that water stress imposed by PEG 4000 causes arrest of cell division in meristematic cells of watermelon seedlings in both G₁ and G₂ periods. PEG and GA treatments resulted in only a partial and transitory synchronization of the cell cycle.     

Elevated plasma proinsulin/insulin ratio is a marker of reduced insulin secretory capacity in healthy young men.

AIM: To examine whether reduced insulin secretory capacity or increased insulin secretory demand is associated with elevated ratio of plasma proinsulin to immunoreactive insulin (PI/IRI ratio) in non-diabetic subjects. SUBJECTS AND METHODS: We measured various indices of insulin secretory function and insulin sensitivity by frequently sampled intravenous glucose tolerance test (FSIGT) and hyerglycemic glucose clamp in 21 healthy young men. We then examined the relationships between these indices and PI, IRI, or PI/IRI ratio in the fasting state. RESULTS: Insulin sensitivity index (SI) measured by FSIGT correlated inversely with basal IRI (r=-0.53, P < 0.01) and PI levels (r=-0.57, P < 0.01), but there was no significant correlation between SI and PI/IRI ratio (r=0.26, NS). On the other hand, PI/IRI ratio correlated inversely with insulin secretory indices, such as acute insulin responses during FSIGT (r =-0.46, P < 0.01) and hyperglycemic glucose clamp (r=-0.54, P < 0.01) and submaximum insulin response during hyperglycemic glucose clamp (r=-0.59, P < 0.01). CONCLUSIONS: These results indicate that elevated PI/IRI ratio may serve as a marker of reduced insulin secretory function in non-diabetic subjects.     

Effects of temperature on the specific dynamic action of the southern catfish, Silurus meridionalis

Specific dynamic action (SDA), the energy expended on all physiological processes that is associated with meal digestion and assimilation, is strongly affected by temperature. We assessed the effects of temperature on the postprandial metabolic response and calculated SDA of the southern catfish, Silurus meridionalis. The fish was fed with experimental diets at a meal size of 4% body mass, and by using an 8-chamber, continuous-flow respirometer the oxygen consumption rate was determined at a 2 h interval until the postprandial oxygen consumption rate returning to the preprandial level, at four different temperatures. The energy expended on SDA (SDA(E)) were 2.71, 3.07, 3.16, and 3.62 kJ, the SDA(coefficients) (energy expended on SDA quantified as a percentage of the digestible energy content of the meal) were 7.70, 9.44, 10.36, and 11.12%, and the peak metabolic rates (R(peak)) of SDA were 3.48, 4.31, 5.96, and 7.30 mg O2 h(-1), at 17.5, 22.5, 27.5, and 32.5 degrees C respectively. The relationships between those parameters and temperature were: SDA(E)=1.74+0.0559T (n=26, r(2)=0.676), SDA(coefficient)=4.10+0.223T (n=26, r(2)=0.726), and R(peak)=-1.34+0.264T (n=26, r(2)=0.896). The SDA durations showed a slow-fast-slow tendency of decrease with increasing temperature, and were 88.00, 85.71, 67.71, and 66.50 h at 17.5, 22.5, 27.5 and 32.5 degrees C respectively. Two separate peaks appeared during the SDA response at 17.5 degrees C, and it might be due to a rapid startup of the mechanical process with a lag of the biochemical process, which suggested that the peaks of "mechanical component" and "biochemical component" of SDA might be separated when temperature was low enough.     

Effects of temperature on the specific dynamic action of the southern catfish, Silurus meridionalis.

Specific dynamic action (SDA), the energy expended on all physiological processes that is associated with meal digestion and assimilation, is strongly affected by temperature. We assessed the effects of temperature on the postprandial metabolic response and calculated SDA of the southern catfish, Silurus meridionalis. The fish was fed with experimental diets at a meal size of 4% body mass, and by using an 8-chamber, continuous-flow respirometer the oxygen consumption rate was determined at a 2 h interval until the postprandial oxygen consumption rate returning to the preprandial level, at four different temperatures. The energy expended on SDA (SDA(E)) were 2.71, 3.07, 3.16, and 3.62 kJ, the SDA(coefficients) (energy expended on SDA quantified as a percentage of the digestible energy content of the meal) were 7.70, 9.44, 10.36, and 11.12%, and the peak metabolic rates (R(peak)) of SDA were 3.48, 4.31, 5.96, and 7.30 mg O2 h(-1), at 17.5, 22.5, 27.5, and 32.5 degrees C respectively. The relationships between those parameters and temperature were: SDA(E)=1.74+0.0559T (n=26, r(2)=0.676), SDA(coefficient)=4.10+0.223T (n=26, r(2)=0.726), and R(peak)=-1.34+0.264T (n=26, r(2)=0.896). The SDA durations showed a slow-fast-slow tendency of decrease with increasing temperature, and were 88.00, 85.71, 67.71, and 66.50 h at 17.5, 22.5, 27.5 and 32.5 degrees C respectively. Two separate peaks appeared during the SDA response at 17.5 degrees C, and it might be due to a rapid startup of the mechanical process with a lag of the biochemical process, which suggested that the peaks of "mechanical component" and "biochemical component" of SDA might be separated when temperature was low enough.     

太白红杉群落物种多样性与环境因子的关系

定量研究了太白红杉群落物种多样性与环境因子的关系,结果表明:(1)海拔与物种丰富度和种间相遇机率呈极显著的负相关;与Shannon指数、Simpson指数和Pielou指数表现为"中间高度膨胀"的规律,即中等海拔高度上多样性高而低海拔和高海拔多样性较低。(2)岩石裸露度与物种丰富度呈显著的负相关。(3)土壤含水量与物种丰富度呈极显著的正相关;与种间相遇机率呈先降后升的趋势。(4)环境因子与物种多样性逐步回归结果是:海拔和岩石裸露度与物种丰富度关系密切,其回归方程为:S=90.62-0.02E-14.14B(r=0.769,P<0.05)。     

Improved indices of insulin resistance and insulin secretion for use in genetic and population studies of type 2 diabetes mellitus.

Homeostasis model assessment (HOMA) provides indices of insulin secretion (beta) and insulin resistance (R) derived from fasting plasma glucose (FPG) and fasting plasma insulin (FPI) levels. However, these indices could not account for a significant heritability of fasting plasma glucose (FPG) (h2 = 0.75, P<0.01) in a group of 214 female twins. This result is consistent with a misclassification between effects due to insulin secretion and resistance in the HOMA indices. We report here evidence of such misclassification in the HOMA indices and describe a minor modification to the model which corrects it. Direct measures of insulin resistance (euglycaemic clamp) and secretion (i.v. glucose bolus) were obtained in 43 non-diabetic subjects. Heritability was estimated by statistical modelling of genetic and environmental influences in data from 214 non-diabetic female subjects. Modified HOMA (HOMA') indices were obtained from beta' = (Ln(FPI) - c)/FPG and R' = (Ln(FPI) - c)*FPG where c is a constant derived from regression analysis of Ln(FPI) vs FPG. Indices from both models correlated with the direct measures similarly (r = 0.63 (R), 0.49 (R'), 0.45 (beta), 0.39 (beta'), all P< 0.01). Directly measured insulin resistance and secretion were not significantly correlated (r = 0.13, P = 0.21). However, unmodified HOMA-beta and R were strongly related (r = 0.78, P<0.0001 vs. 0.13) demonstrating substantial misclassification. The relationship between beta' and R' (r = 0.13) was not different from that between the two direct measures and significant heritability of beta' (h2 = 0.68, P<0.01) and R' (h2 = 0.59, P<0.05) was evident in the twin data. The proposed modification to HOMA significantly reduces misclassification and reveals separate components of insulin resistance and insulin secretion in the heritability of FPG.     

Serum Vaspin Levels Are Associated with the Development of Clinically Manifest Arthritis in Autoantibody-Positive Individuals

Objectives

We have previously shown that overweight may increase the risk of developing rheumatoid arthritis (RA) in autoantibody positive individuals. Adipose tissue could contribute to the development of RA by production of various bioactive peptides. Therefore, we examined levels of adipokines in serum and synovial tissue of subjects at risk of RA.

Methods

Fifty-one individuals positive for immunoglobulin M rheumatoid factor (IgM-RF) and/or anti-citrullinated protein antibodies (ACPA), without arthritis, were included in this prospective study. Levels of adiponectin, vaspin, resistin, leptin, chemerin and omentin were determined in baseline fasting serum samples (n = 27). Synovial tissue was obtained by arthroscopy at baseline and we examined the expression of adiponectin, resistin and visfatin by immunohistochemistry.

Results

The development of clinically manifest arthritis after follow-up was associated with baseline serum vaspin levels (HR1.5 (95% CI 1.1 to 2.2); p = 0.020), also after adjustment for overweight (HR1.7 (95% CI 1.1 to 2.5); p = 0.016). This association was not seen for other adipokines. Various serum adipokine levels correlated with BMI (adiponectin r = -0.538, leptin r = 0.664; chemerin r = 0.529) and systemic markers of inflammation such as CRP levels at baseline (adiponectin r = -0.449, omentin r = -0.557, leptin r = 0.635, chemerin r = 0.619, resistin r = 0.520) and ESR (leptin r = 0.512, chemerin r = 0.708), p-value<0.05. Synovial expression of adiponectin, resistin and visfatin was not associated with development of clinically manifest arthritis.

Conclusions

In this exploratory study, serum adipokines were associated with an increased inflammatory state in autoantibody-positive individuals at risk of developing RA. Furthermore, serum vaspin levels may assist in predicting the development of arthritis in these individuals.     

Thermodynamics of the Binding of Chymotrypsin with the Black-eyed Pea Trypsin and Chymotrypsin Inhibitor (BTCI)

The binding of -chymotrypsin to black-eyed pea trypsin/chymotrypsin inhibitor (BTCI) has been studied using the inhibitory activity against the enzyme and the formation of the complex enzyme/inhibitor followed by measurements of fluorescence polarization. Apparent equilibrium constants were estimated for several temperatures and the values obtained range from 0.32 × 10⁷ to 1.36 × 10⁷ M^–1. The following values were found from van't Hoff plots: H _vh ^° = 10.8 kcal mol^-1 (from inhibitory assays) and 11.1 kcal mol^–1 (from fluorescence polarization); S° = 67.9 and = 67.8 kcal K^–1 mol^–1, respectively. Calorimetric binding enthalpy was determined (corrected for the ionization heat of the buffer) and the resulting value was H _cal ^° = 4.9 kcal mol^-1. These results indicate that the binding of chymotrypsin to BTCI is an entropically driven process.