Combined segregation and linkage analysis of genetic hemochromatosis using affection status, serum iron, and HLA.

Characterizing the distribution of parameters of iron metabolism by hemochromatosis genotype remains an important goal vis-à-vis potential screening strategies to identify individuals at genetic risk, since a specific marker to detect the abnormal gene has not been identified as yet. In the present investigation, we analyze serum iron values in ascertained families using a method which incorporates both segregation of the clinical affection status and the HLA linkage information to identify the underlying genotypes. The analysis is performed using an extension of the model presented by Bonney et al., comprising regressive models for segregation analysis and the multipoint linkage strategy implemented in LINKAGE. The gene was found to be completely recessive with respect to both clinical manifestations and serum iron abnormalities, with significant differences in expression by sex. Clinical manifestations were present for all male homozygotes in this data set, suggesting that the recessive hemochromatosis genotype is fully penetrant at all ages in males. This was not the case for younger females. Significant genotype-specific age and sex effects were found for serum iron values. It is interesting that deletion of the HLA marker information did not affect our ability to resolve the genetic model when we analyzed a bivariate phenotype. This serves as a reminder that a search for relevant biological markers can be equally important in discerning the genetic etiology of a disease trait, as a search for linked genetic markers.     

Serum ferritin as a marker of affection for genetic hemochromatosis

A bivariate segregation analysis of genetic hemochromatosis with serum ferritin concentration was undertaken to examine the pleiotropic effect of the hemochromatosis locus on each of the two phenotypes, in an ascertained sample of families from Brittany, France. The gene was recessive with respect to both phenotypes, and the estimated gene frequency in the general population was 0.054. Although the ferritin concentration was corrected for the linear relationship with age among controls, there was a residual correlation with age among male family members, consistent with the progressive increase in body iron stores among hemochromatosis homozygotes. This genotype-specific relationship with age illustrates the importance of incorporating interaction effects into analytic models, and suggests that even as a better indicator of progress of disease, rather than liability to disease, serum ferritin concentration serves well to distinguish hemochromatosis homozygotes from alternate genotypes in a family study.     

Usefulness of erythrocyte ferritin analysis in hereditary hemochromatosis.

A study was carried out to determine the usefulness of erythrocyte ferritin analysis in identifying homozygotes and heterozygotes in families affected with hereditary hemochromatosis, an autosomal recessive disorder. To select the subjects the genotypes of 60 people from 26 affected families were determined by HLA-A and HLA-B haplotyping. In addition, data for 12 homozygotes for whom erythrocyte ferritin values were available from the literature were included. Likelihood analysis was used to evaluate the diagnostic value of erythrocyte ferritin analysis alone and in combination with serum ferritin testing. An erythrocyte ferritin value of 150 ag/cell or higher combined with a serum ferritin level above the 90th percentile indicated homozygosity, whereas a value of less than 150 ag/cell and a serum ferritin level at or below the 90th percentile indicated that homozygosity could be ruled out with a high degree of confidence. The probability of heterozygosity rose to 92% when the erythrocyte ferritin value was between 29 and 149 ag/cell and to 98% when this result was combined with a serum ferritin level at or below the 90th percentile. Erythrocyte ferritin analysis in combination with serum ferritin testing is useful for identifying homozygotes and a proportion of heterozygotes in families affected with hemochromatosis.     

Regulation of immunocompetence by different androgen metabolites in a blenny with alternative reproductive tactics

In Parablennius parvicornis, small reproductive males with relatively low expression of secondary sexual characters (M- morphotype) parasite on the parental investment of the larger nest-holder males which have fully developed secondary sexual characters (M+ morphotype). In comparison with M+ males, M- males have relatively low levels of androgens while having high blood cell percentages of lymphocytes and antigen responsiveness. Here we test the hypothesis that androgens are a causal factor for these differences in immunocompetence between morphotypes. After drawing an initial blood sample, males received a silastic implant containing either oil only (C), or oil with testosterone (T) or 11-ketotestosterone (KT). Males were re-caught 2 weeks later for drawing of the final blood sample. KT but not T induced the development of secondary sexual characters in M- males. M- males treated with KT showed lower swimming activity than the males treated with T or C implants, suggesting that KT also mediates behavioral changes in M- males. As expected, blood cell percentages of lymphocytes, but not of granulocytes, were higher in M- males than in M+ males. Overall, lymphocyte percentages increased in the C group which might have been a response to the surgery/treatment. In concordance with the hypothesis, lymphocyte percentages were suppressed in males treated with T in comparison with controls. However, no significant change was found in KT-treated males. This suggests that androgens modulate central, morphological and immunological traits by partly independent androgen mechanisms in P. parvicornis.     

Serum insulin-like growth factor I reference values for an automated chemiluminescence immunoassay system: results from a multicenter study

BACKGROUND: Analysis of insulin-like growth factor I in serum (S-IGF-I) is an integral component in the diagnosis of GH-related disorders and is going to be of interest in the diagnosis and follow-up of many disorders. The objective of the present study was to develop cross-sectional reference values for S-IGF-I measured by an automated chemiluminescence immunoassay (Nichols Advantage). METHODS: The study included samples from 3,961 healthy subjects (2,201 males, 1,760 females) aged 1 month to 88 years. Six laboratories were involved in this study and the samples were analyzed by one of seven automated immunoassay systems run in these laboratories. For data analysis, polynomial age and sex-specific models were fitted after transformation of S-IGF-I values. RESULTS: The results show the well-known age dependency of S-IGF-I levels. At ages <20, higher S-IGF-I levels were seen in girls with an estimated mean peak of 410 microg/l at age 14 and an estimated mean peak of 382 microg/l at age 16 in boys. Thereafter, a rapid decrease was seen to approximately 25 years of age, followed by a slow age-dependent decrease. In adulthood, S-IGF-I in males were slightly, but significantly higher than in females. It could be shown that the mean values of some reference sample subgroups differed significantly from the total mean. However, the multicenter approach used in this study reduces the impact of systematic population, sample handling and laboratory differences on the calculated reference mean. CONCLUSION: The present study establishes age- and sex-specific reference values for a fully automated immunoassay system based on a large population of healthy subjects. The established reference values may be used for this immunoassay system in different laboratories provided that the systematic difference between systems is low.     

A haplotype and linkage disequilibrium analysis of the hereditary hemochromatosis gene region

Hereditary hemochromatosis is a recessive disease of iron metabolism widely distributed among people of European descent. Most patients have inherited the causative mutation from a single ancestor. In the course of cloning the hemochromatosis gene, genotypes were generated for these samples at 43 microsatellite repeat markers that span the 6.5-Mb hemochromatosis gene region. The data used to reconstruct the ancestral haplotype across the hemochromatosis gene region are presented in this paper. Portions of the ancestral haplotype were present on 85% of patient chromosomes in this sample and ranged in size from approximately 500 kb to greater than 6.5 Mb. Only one marker, D6S2239, was identical by descent on all of the patient chromosomes containing the ancestral mutation. In contrast, only 3 of the 128 control chromosomes, or 2.3%, carried the ancestral mutation and the surrounding ancestral haplotype. To test new methods for gene finding using linkage disequilibrium we analyzed the genotypic data with a multilocus maximum likelihood method (DISMULT) and a single point method (DISLAMB), both written to analyze data generated from multi-allelic markers. The maximum value from DISLAMB analysis occurred at marker D6S2239, which is less than 20 kb from the hemochromatosis gene HFE, consistent with the haplotype analysis. The peak of the multi-point analysis was 700 kb from HFE, possibly due to the nonuniform recombination rates within this large region. The recombination rate appears to be lower than expected centromeric of the HFE gene. Received: 10 June 1997 / Accepted: 4 December 1997     

Macrophage iron, hepcidin, and atherosclerotic plaque stability

Hepcidin has emerged as the key hormone in the regulation of iron balance and recycling. Elevated levels increase iron in macrophages and inhibit gastrointestinal iron uptake. The physiology of hepcidin suggests an additional mechanism by which iron depletion could protect against atherosclerotic lesion progression. Without hepcidin, macrophages retain less iron. Very low hepcidin levels occur in iron deficiency anemia and also in homozygous hemochromatosis. There is defective retention of iron in macrophages in hemochromatosis and also evidently no increase in atherosclerosis in this disorder. In normal subjects with intact hepcidin responses, atherosclerotic plaque has been reported to have roughly an order of magnitude higher iron concentration than that in healthy arterial wall. Hepcidin may promote plaque destabilization by preventing iron mobilization from macrophages within atherosclerotic lesions; the absence of this mobilization may result in increased cellular iron loads, lipid peroxidation, and progression to foam cells. Marked downregulation of hepcidin (e.g., by induction of iron deficiency anemia) could accelerate iron loss from intralesional macrophages. It is proposed that the minimally proatherogenic level of hepcidin is near the low levels associated with iron deficiency anemia or homozygous hemochromatosis. Induced iron deficiency anemia intensely mobilizes macrophage iron throughout the body to support erythropoiesis. Macrophage iron in the interior of atherosclerotic plaques is not exempt from this process. Decreases in both intralesional iron and lesion size by systemic iron reduction have been shown in animal studies. It remains to be confirmed in humans that a period of systemic iron depletion can decrease lesion size and increase lesion stability as demonstrated in animal studies. The proposed effects of hepcidin and iron in plaque progression offer an explanation of the paradox of no increase in atherosclerosis in patients with hemochromatosis despite a key role of iron in atherogenesis in normal subjects.     

Study of somatotype in Italian children aged 6 to 10 years

The aim of the present research was to study the variations of somatotype, calculated by the Heath-Carter anthropometric technique, during growth in a sample of children (416 males and 402 females), aged 6 and 10 years, attending primary and secondary schools of L'Aquila and its province (Abruzzo, Italy). The sample was subdivided into “urban” and “non-urban” groups, on the basis of the residence of the children, to examine possible differences in growth related to the different environments. This study give an account of the somatotype components between urban and non urban childreen between the age 6 and 10 years. A tendency toward an increase of endomorphy (adipose component) with age was noticed in both sexes. In females, ectomorphy (component of physical linearity) tended to increase and mesomorphy (muscular-skeletal component) showed a slight decrease during growth, while males exhibited a discontinuous trend. The differences between urban and non-urban children were not significant, although generally higher values of endomorphy and mesomorphy were found in males and females of the urban sample. The differences between the sexes consisted of higher values of endomorphy and lower values of mesomorphy in females. Ectomorphy was similar in the two sexes.     

闽南人的体质特征

在福建漳州地区调查了322例男性(城市男性为149例,乡村汉族男性为173例)和357例女性(城市女性为164例,乡村女性为193例)成人的86项体质指标,计算了24项体质指数,统计了指数分型情况,与我国族群资料进行了比较,对闽南人体质特征进行了初步分析。结果 1)闽南人有皱褶率高,有蒙古褶率低,眼裂高度眼裂多为狭窄型,眼外角多高于眼内角,鼻根高度多为中等型,直鼻背,颧部突出度多为微弱型;鼻基部男性多呈水平,女性多呈上翘;鼻翼高度多为中等,鼻孔最大径多为斜位,鼻翼较宽;男性耳垂多为圆形,女性多为三角形;上唇皮肤部高度多为中等,红唇男性薄型率、中等型均较高,女性薄型率最高,发黑,肤黄、眼褐。2)总体说来,闽南人头面部测量指标值介于北亚、南亚类型族群之间。3)从指数均数来看,闽南人男女性均为圆头型、高头型、狭面型,狭鼻型。城市女性为狭头型,城市男性、乡村男性、乡村女性为中头型。闽南人头面部指数值更接近于南亚类型族群。4)从体部指标值和体部指数来看。闽南人更接近于北亚类型族群。闽南人体质介于南亚与北亚类型族群之间,男性相对接近于北亚类型族群,女性相对接近于南亚类型族群。     

Effect of pH on the stability of hemochromatosis factor E: a combined spectroscopic and molecular dynamics simulation-based study

Hereditary hemochromatosis is an iron overburden condition, which is mainly governed by hereditary hemochromatosis factor E (HFE), a member of major histocompatibility complex class I. To understand the effect of pH on the structure and stability of HFE, we have cloned, expressed, and purified the HFE in the bacterial system and performed circular dichroism, fluorescence, and absorbance measurements at a wide pH range (pH 3.0–11.0). We found that HFE remains stable in the pH range 7.5–11.0 and gets completely acid denatured at low pH values. In this work, we also analyzed the contribution of salt bridges to the stability of HFE. We further performed molecular dynamics simulations for 80 ns at different pH values. An excellent agreement was observed between results from biophysical and MD simulation studies. At lower pH, HFE undergoes denaturation and may be driven toward a degradation pathway, such as ubiquitination. Hence, HFE is not available to bind again with transferrin receptor1 to negatively regulate iron homeostasis. Further we postulated that, might be low pH of cancerous cells helps them to meet their high iron requirement.     

Consensus DNA site for the Escherichia coli catabolite gene activator protein (CAP): CAP exhibits a 450-fold higher affinity for the consensus DNA site than for the E. coli lac DNA site.

We have synthesized two 40 base pair DNA fragments; one fragment contains the consensus DNA site for CAP (fragment 'ICAP'); the other fragment contains the E. coli lac promoter DNA site for CAP (fragment 'LCAP'). We have investigated the binding of CAP to the two DNA fragments using the nitrocellulose filter binding assay. Under standard conditions [( NaCl] = 200 mM, pH = 7.3), CAP exhibits a 450-fold higher affinity for ICAP than for LCAP. The salt dependence of the binding equilibrium indicates that CAP makes eight ion pairs with ICAP, but only six ion pairs with LCAP. Approximately half of the difference in binding free energy for interaction of CAP with ICAP vs. LCAP is attributable to this difference in ion-pair formation. The pH dependence of the binding equilibrium indicates that the eight CAP-ICAP ion pairs and the six CAP-LCAP ion pairs do not involve His residues of CAP.     

Assessing mating performance of male Glossina pallidipes (Diptera: Glossinidae) using a walk-in field cage

To monitor the quality of male tsetse for use in the sterile insect technique (SIT), a field cage test was developed and evaluated. Mating competitiveness was tested with male Glossina pallidipes Austen that emerged from pupae stored for different periods at 15 degrees C. Control males emerged from pupae stored at 23-24 degrees C and emerged at 26.5 degrees C. Each sample of test males was divided into two groups with one group being irradiated at 120 Gy; the other group was not irradiated. More than 70% of the maximum possible number of mating pairs occurred in all tests. Males emerged from pupae kept at low temperature and then irradiated formed a greater proportion of mating pairs than the controls. Males emerged from pupae kept at 15 degrees C generally started mating more quickly than the standard colony males although there was no significant difference. Insemination rates were above 99%. Pooled data indicated that mean spermathecal values for females mated with irradiated males were significantly lower than for control males. The duration of copulation varied significantly between treatment groups and was significantly longer for irradiated male flies; there was no correlation between duration of copulation and mean spermathecal value.     

Biomechanical beam analysis of long bones from a late 18th century slave cemetery in Cape Town, South Africa

This study aims to quantify the physical demands of a sample of late 18th century skeletons from an unmarked burial site on Cobern Street, Cape Town, South Africa. Historical studies suggest that these individuals were either slaves or "free black" people of low socioeconomic standing. Cortical cross-sectional areas of paired humeri and tibiae from the Cobern Street collection (N = 29), a modern cadaver collection (N = 31), and a hunter-gatherer collection (N = 30) were compared by means of biomechanical beam analysis on computerized tomography scans. Results showed that the Cobern Street sample, both males and females, were closer to the modern group in total tibial cortical area and in the second moments and polar moments of cortical area, than to the hunter-gatherer group. It is assumed that these results can be explained by differences in lower limb activity. Tibial strength properties showed the hunter-gatherer peoples to be highly mobile and active walkers in comparison to the Cobern Street and modern samples. The males from the Cobern Street sample showed significantly higher values for humeral strength properties than either the hunter-gatherer or modern individuals, attesting to their status as manual laborers. The humeral cross-sectional strength properties for females were very similar between the Cobern Street and modern groups but again significantly different from the hunter-gatherer sample. The domestic chores performed by females of the recent cadaver sample may be very similar to those performed by the Cobern Street sample.     

Coenzyme Q10 and differences in coronary heart disease risk in Asian Indians and Chinese.

Indians or South Asians have been found to be particularly susceptible to coronary heart disease (CHD) in many countries. A novel risk factor for CHD may be coenzyme Q10 (CoQ10). In this study, plasma CoQ10 (including ubiquinol-10, CoQ10H2, and total CoQ10), various lipid parameters, and antioxidant levels were determined in a random sample of Indians and Chinese from the general population of Singapore. The reduced form of coenzyme Q10, CoQ10H2, and total Q10 concentrations in plasma were significantly lower in Indian males than Chinese males. Although no significant differences were found in plasma concentrations of total cholesterol, triglycerides, and low-density lipoprotein cholesterol (LDL) between the two ethnic groups, the ratios of ubiquinol and total CoQ10 to triglycerides, total cholesterol, and LDL were significantly lower in Indian males than Chinese males. There were no significant ethnic differences in other antioxidant levels, including trans-retinol, alpha-tocopherol, and ascorbic acid. The consistently lower values of coenzyme Q10, especially its reduced form, in Indian males may contribute to the higher susceptibility of this ethnic group to coronary heart disease.     

Quantitative finger dermatoglyphics in a Spanish population (Tierra de Campos)

This study deals with the finger ridge counts and the distribution of their frequencies drawn from a Spanish sample consisting of 833 school-children (417 males and 416 females), which has been collected from the geographical area of Tierra de Campos. Paired t-test and Student-t-test were used to explore bimanual and sexual differences, respectively. The results showed: a bimanual asymmetry marked by significantly higher right hand ridge count for thumb and index pairs in both males and females, ridge counts of males are always higher than the corresponding values of females; the differences being significant excepting for right and left index and for left ring-finger. The frequency distribution of TFRC was slightly, but significantly, different from normality only in males, as Kolmogorov test showed. A great homogeneity between values for TFRC of males and females from Tierra de Campos and those of the available Spanish and Portuguese populations has been found, the values being high not only in the variation ranges of the Spanish and Portuguese populations, but also in the ranges reported for other European populations.     

Endocrine patterns before and after examination stress in males and females

Plasma levels of cortisol, adrenocorticotropin (ACTH), and growth hormone (GH), were determined by radioimmunoassay in medical students just before (Exp.1) plasma values of cortisol were higher than their control values in both sexes, statistically significantly, however, only in males. The difference of cortisol behavior between the sexes was not significant. The males' ACTH values were lower than their control levels, but there was no such difference in the females. Plasma levels and differences of growth hormone varied greatly in both sexes. After the examination (Exp.2) plasma values of cortisol were unaffected compared with their control values in both sexes. The ACTH values were lower than the control values in both sexes, significantly though only in the males. The GH values were higher than control values in the males but not in the females. The results are in line with previous observations suggesting that psychological stress is associated with slightly different hormonal effects in males and females.     

Blood pressure in young adults as influenced by copper and zinc intake

The relationship of copper and zinc status with blood pressure in young adults was studied (N=59). Copper and zinc status was assessed through analysis of serum, urine, hair, and diet records. Males (N=27) had greater systolic and pulse pressures than females (N=32). Multiple regression analysis suggested that males and higher caloric and dietary copper intakes were the predominant factors associated with higher systolic and diastolic pressures, whereas higher dietary zinc intakes were associated with lower pressures. Higher urine zinc excretion rates and serum zinc concentrations were associated with higher diastolic pressures. The sex variable was the strongest factor affecting pulse pressures, with males having higher pulse pressures. Serum zinc concentrations had an inverse relationship to pulse pressures. The data suggest that the weight/height index and age had a low association with blood pressure in this normotensive sample.     

Hematologic Values of Free-ranging Cebus cay and Cebus nigritus in Southern Brazil

We evaluated blood samples obtained from 80 free-ranging healthy capuchins (Cebus cay and C. nigritus) to establish hematological reference values and to assess the influence of sex and age on them. We caught the monkeys in the Paraná River region of Southern Brazil via manual or automatic traps. We anesthetized them intramuscularly with 3.6 mg/kg tiletamine/zolazepam hydrochlorides. After physical examinations, we divided the sample according to sex and age: 26 females (13 adults and 13 juveniles) and 54 males (27 adults and 27 juveniles). We collected blood and determined hematological values via traditional published methods. We analyzed data via 2-way ANOVA to test the effect of sex, age, and interactions between the 2 factors. The packed-cell volume was higher in adult males and the numbers of white blood cells and lymphocytes were higher in juveniles. There is no other significant difference.     

Rearing management of rabbit males selected by high growth rate: the effect of diet and season on semen characteristics

A total of sixty-six young males were used to evaluate the effect of low (L), medium (M) and high (H) concentrations of dietary digestible energy received during the rearing seasons (autumn and spring) on the performance and main semen characteristics of males for artificial insemination selected by a high growth rate. Males reared during the spring season presented a significantly higher weight at weaning than those reared during the autumn season (P < 0.001), and these differences were maintained until the end of the trial. The requirements of the males were easily covered as a general rule. In the autumn group, the males were unable to intake the digestible protein recommended only during their 3rd month of life, especially with low concentrate diets (P < 0.05). H males showed higher semen concentration and production during the autumn season, while L males showed a higher semen concentration and production than M males during the spring season, the H group showed intermediate values (P < 0.001). Males reared during the spring season showed significantly higher values of sperm concentration (P < 0.01) and production (P < 0.01). H males presented a lower percentage of spermatozoa with cytoplasmic droplets than the L group (P < 0.05) and the lowest values for sperm abnormalities during the autumn season, while the L group presented higher values for percentage abnormalities, especially during the last month controlled (P < 0.05). As a general rule, the main motility parameters controlled were not affected by the rearing diet received nor the season. These results seem to indicate that the management of rabbit males during the growing and rearing periods seem to significantly affect their subsequent performance and semen production.     

Body shape in relation to socio-economic status in young adults from the basque Country

Body shape has a great variability determined, partly, by energy intake and physical activity, as well as by gender and age. The aim of this research was to analyse the relation between socio-economic status (SES) and body shape estimated through the somatotype, in a sample of university students. The sample included 316 males and 635 females aged 18-33. Somatotype was estimated by the Heath-Carter anthropometric technique. The information included data of SES. A MANOVA test was used to test differences among groups. The mean somatotypes were 4.0-4.4-2.3 in males and 5.4 -3.4-2.2 in females. Sexual dimorphism for the whole somatotype was found (p < 0.001). There is a greater stability of the body shape in females regarding SES and more variability in males. Males' somatotype was significantly related to SES, low socioeconomic levels were associated with high values of endomorphy and mesomorphy.