Human centromere protein C (CENP-C) is a DNA-binding protein which possesses a novel DNA-binding motif.

Mammalian centromere proteins (CENPs) can be divided into those that translocate from centromere to midzone in the progress of mitosis, and those that remain at the centromere throughout the cell cycle. The latter including CENP-A, CENP-B, and CENP-C is the candidate for DNA-binding protein. CENP-B has been shown previously to possess the specific DNA-binding activity to 17-base pair sequences dispersed on human centromeric alphoid repeats. In this study, we examined DNA-binding property of CENP-C that is localized to inner kinetochore plate of the metaphase chromosome. We independently isolated a full-length cDNA encoding human CENP-C and expressed it as the polypeptide tagged with histidine oligomer in Escherichia coli. After affinity purification with Ni(2+)-chelated resin, DNA-binding activity of the recombinant CENP-C renatured on the membrane was demonstrated by using human genomic DNA and an alphoid subfamily in South-Western-type blotting analysis. By constructing a series of truncated products, the DNA-binding domain was located at an internal 101-amino-acid stretch with no apparent homology to any other DNA-binding proteins. This may suggest that CENP-C is directly involved in formation of kinetochore chromatin fibers.

Lack of correlation between different measurements of proprioception in the knee.

Current methods of measurement of proprioceptive function depend on the ability to detect passive movement (kinaesthesia) or the awareness of joint position (joint position sense, JPS). However, reports of proprioceptive function in healthy and pathological joints are quite variable, which may be due to the different methods used. We have compared the validity of several frequently used methods to quantify proprioception. Thirty healthy subjects aged between 24 and 72 years underwent five established tests of proprioception. Two tests were used for the measurement of kinaesthesia (KT1 and KT2). Three tests were used for the measurement of JPS, a passive reproduction test (JPS1), a relative reproduction test (JPS2) and a visual estimation test (JPS3). There was no correlation between the tests for kinaesthesia and JPS or between the different JPS tests. There was, however, a significant correlation between the tests for kinaesthesia (r = 0.86). We conclude therefore that a subject with a given result in one test will not automatically obtain a similar result in another test for proprioception. Since they describe different functional proprioceptive attributes, proprioceptive ability cannot be inferred from independent tests of either kinaesthesia or JPS.

Mapping of the human CENP-B gene to chromosome 20 and the CENP-C gene to chromosome 12 by a rapid cycle DNA amplification procedure.

By optimizing the primer-annealing temperature in a rapid air cycling procedure, two human DNA sequences encoding centromere proteins B and C (CENP-B and CENP-C) were specifically amplified without any detectable amplification of highly homologous rodent DNA sequences. Using a panel of rodent/human hybrid DNA, the genes for human CENP-B and CENP-C were conveniently mapped to chromosomes 20 and 12, respectively.

A cross-sectional study of the size and strength of the lower leg muscles during growth.

The influences of age and sex on the cross-sectional area (CSA) and isometric strength of the ankle dorsiflexors and plantarflexors (PF) were investigated in four age groups of 121 boys and 121 girls aged: 7-9, 10-12, 13-15, and 16-18 years. A single anatomical cross-section was determined at 30% of the distance from the articular cleft between the femur and tibiacondyles by using an ultrasonic apparatus. In both sexes, the increase in age was associated with significant increases in the CSA and strength (ST) of these opposing muscle groups. The sex differences became apparent in the 13-15 year group for CSA and in the 16-18 year group for ST but the differences reduced considerably when CSA and ST were expressed per unit of the second power of the lower leg length (CSA.LL-2) and the product of CSA and the lower leg length (ST.CSA-1.LL-1), respectively. However, CSA.LL-2 of both muscles had a tendency to be increased at and over the age of 10-12 years, and was the highest at 16-18 years, and ST.CSA-1.LL-1 of PF showed higher values in the older boys than in the younger. Thus, it appeared that, at least in the reciprocal muscle groups of the ankle joint, the sex differences in muscle CSA and ST during growth could be accounted for by differences in LL and muscle mass, respectively. However, other factors must also be involved to explain completely the age differences in these variables.

An antigenic determinant on human centromere protein B (CENP-B) available for production of human-specific anticentromere antibodies in mouse.

Centromere protein B (CENP-B) is one of the centromere DNA binding proteins constituting centromere heterochromatin throughout the cell cycles. Some components of mammalian centromeres including CENP-B are target antigens for autoimmune disease patients, often those with scleroderma. Recent isolations of CENP-B genes from human and mouse suggested that CENP-B was highly conserved among mammals. From the previous analysis of the reactivity of patient anticentromere sera, two autoepitopes have been located on the DNA binding domain at the amino-terminal region. The amino acid sequences for both the epitopes are perfectly conserved in the two species, human and mouse. In this study, to identify a human-specific antigenic determinant, the remaining two epitopes were further located in separate carboxyl-terminal regions of human CENP-B. Although the amino acid sequence of one epitope is identical to that of the corresponding region in mouse CENP-B, the other has a less homologous sequence. To confirm that the latter epitope was available for production of human-specific anticentromere antibodies, mice were immunized with the recombinant human CENP-B product. One serum that exclusively stained human centromere structure, but not that of other mammals, was identified in the immunofluorescence microscopic observation. The epitope analysis showed that the less conserved one was recognized by this serum. These results suggested that the corresponding region defines the antigenic determinants for the species specificity.

Changes in force-velocity and power output of upper and lower extremity musculature in young subjects following 20 days bed rest.

Ten healthy sedentary students were exposed to 20 days bed rest (BR) to study the effect of simulated weightlessness on force(F)-velocity(V) characteristics and power(P) output of upper and lower limb movements. F, V and P were measured using a special dynamometer applicable to single joint movements [elbow flexion(EF) and extension(EE), knee flexion(KF) and extension(KE), and hip flexion] or multi-joint movements (squatting). Physiological cross-sectional areas(PCSA) of KF and KE muscles were measured by a magnetic resonance imaging technique. After BR, attenuation in P were observed in lower limb movements (decreased by 19.8-43.6% for squatting, KF and KE), in upper limb movements on the other hand, the decreases in P were not significant (approximately -5% for EF and EE). Decrease in P in lower limb were more pronounced in heavier loaded conditions which were characterized by decreases in both F and V. For KF and KE, decreases in maximal static F (-18.9 to approximately -26.8%) were more pronounced than the decreases observed in the PCSA (approximately -7%), resulting in decreases in specific tension (-12.3 to approximately -22.1%). Neural excitation potentials to generate maximal muscle tension or P might also be influenced by weightlessness.

Relation between grip strength and radial bone mineral density in young athletes.

In this study, we evaluated the relationship between bone mineral density (BMD) and muscle strength in young athletes who had not yet experienced age-related bone loss. Radial BMD and grip strength were measured in 10 male college wrestlers, 16 female college basketball players, and 12 female college tennis players. Radial BMD was measured in the distal and middle radius by dual energy x-ray absorptiometry (DEXA). Isometric grip strength was assessed with a hand-held dynamometer. The dominant forearm was examined in the amateur wrestlers and basketball players for grip strength and BMD. Both forearms were examined in the tennis players. A significant positive correlation was found between radial BMD and grip strength in the dominant forearm, and between radial BMD and body weight. Moreover, to eliminate a possible effect of body weight on radial BMD, we compared radial BMD with grip strength in both the dominant and nondominant arm of 12 college tennis players. Grip strength in the dominant forearm was significantly greater than in the nondominant forearm. The midradial BMD of the dominant forearm was also significantly higher than in the nondominant forearm. Based on these findings, we conclude that grip strength is one of the determinant factors of radial BMD in the dominant forearm of young college athletes.