Height growth velocity, islet autoimmunity and type 1 diabetes development: the Diabetes Autoimmunity Study in the Young.

AIMS/HYPOTHESIS: Larger childhood body size and rapid growth have been associated with increased type 1 diabetes risk. We analysed height, weight, BMI and velocities of growth in height, weight and BMI, for association with development of islet autoimmunity (IA) and type 1 diabetes. METHODS: Since 1993, the Diabetes Autoimmunity Study in the Young (DAISY) has followed children at increased type 1 diabetes risk, based on HLA-DR, -DQ genotype or family history, for the development of IA and type 1 diabetes. IA was defined as the presence of autoantibodies to insulin, GAD or protein tyrosine phosphatase islet antigen 2 twice in succession, or autoantibody-positive on one visit and diabetic at the next consecutive visit within 1 year. Type 1 diabetes was diagnosed by a physician. Height and weight were collected starting at age 2 years. Of 1,714 DAISY children <11.5 years of age, 143 developed IA and 21 progressed to type 1 diabetes. We conducted Cox proportional hazards analysis to explore growth velocities and size measures for association with IA and type 1 diabetes development. RESULTS: Greater height growth velocity was associated with IA development (HR 1.63, 95% CI 1.31-2.05) and type 1 diabetes development (HR 3.34, 95% CI 1.73-6.42) for a 1 SD difference in velocity. CONCLUSIONS/INTERPRETATION: Our study suggests that greater height growth velocity may be involved in the progression from genetic susceptibility to autoimmunity and then to type 1 diabetes in pre-pubertal children.

Cerebellar volume in humans related to magnitude of classical conditioning.

Neural circuits in the cerebellum are essential for eyeblink classical conditioning, and hippocampal activation is also present during acquisition. Anatomical (volumetric) brain MRI, delay eyeblink conditioning and neuropsychological tests were administered to eight healthy older subjects. The correlation between cerebellar volume (corrected for total cerebral volume) and conditioned response percentage was 0.81 (p < 0.02), but neither hippocampal nor total cerebral volume correlated with conditioning or any neuropsychological test scores. There was no relationship between age and cerebellar volume, but the correlation between hippocampal volume and age was -0.80 (p < 0.02). These volumetric results add to the increasing evidence in humans demonstrating a relationship between the integrity of the cerebellum and eyeblink classical conditioning.

Anterior cruciate ligament injuries in female athletes: why are women more susceptible?

Anterior cruciate ligament (ACL) injuries occur most frequently in planting and cutting sports such as basketball, soccer, and volleyball. National Collegiate Athletic Association injury data show that female athletes injure the ACL more frequently than their male counterparts do. The greater incidence of ACL injuries in women probably stems from complex, interrelated factors, possibly including hamstring-quadriceps strength imbalances, joint laxity, and the use of ankle braces. Successful treatment often includes surgery.

Association of fragile X syndrome with delayed replication of the FMR1 gene.

The fragile X syndrome is commonly associated with mutant alleles of the FMR1 gene that are hypermethylated and have large expansions of CGG repeats. We present data here on the replication timing of FMR1 that confirm predictions of delayed replication of alleles from affected males. The normal FMR1 allele replicates late in S phase, while alleles from affected males replicate later, the major peak of replication occurring in the flow cytometry fraction usually referred to as G2/M. The delayed timing of replication is not the direct result of a single replication fork stalling at the expanded CGG repeat, because delayed replication was observed for regions on both sides of the repeat. The domain of altered replication timing includes sites at least 150 kb 5' and 34 kb 3' of the repeat, indicating that genes in addition to FMR1 may be affected.

Two progenitor cells for human oogonia inferred from pedigree data and the X-inactivation imprinting model of the fragile-X syndrome.

Laird has proposed that the human fragile-X syndrome is caused by abnormal chromosome imprinting. The analysis presented here supports and extends this proposal. Using published pedigrees that include DNA polymorphism (RFLP) data, we establish that the states of the fragile-X mutation termed "imprinted" and "nonimprinted" usually can be distinguished by the level of cytogenetic expression of the fragile-X chromosome. This information is then used to assess the state of the fragile-X allele in carrier progeny of individual women who inherited a nonimprinted fragile-X chromosome. From this assessment, an estimate is made of the frequency, in individual women, of primary oocytes with an imprinted fragile-X chromosome. The results of this analysis provide additional support for the specific model in which chromosome imprinting occurs in a female in, on average, half of her primary oocytes. This is the expected frequency if X-chromosome inactivation is the initial step in the imprinting of the mutant fragile-X allele. Moreover, this analysis suggests a biological explanation for peculiarities of fragile-X inheritance described by others as "clustering" and the "Sherman paradox." We interpret these peculiarities as consequences of a very small number of oogonial progenitor cells. Two progenitor cells for oogonia is the best integer estimate of the number of such cells at the time of the initial event that leads to chromosome imprinting.

The ultrastructure of upstream and downstream regions of an active Balbiani ring gene.

When active, the 37 kb Balbiani ring genes are known to form transcription loops with an almost fully extended chromatin axis. Here we examine the upstream and downstream regions of such transcription loops by electron microscopy. We demonstrate that a loop starts and ends in tightly packed chromatin; the two anchoring sites are clearly separated from each other in space. The upstream, nontranscribed region consists of a thin, extended, apparently flexible and nucleosome-free fiber corresponding to about 0.5 kb DNA. The downstream, nontranscribed region appears as a 200 nm long nucleofilament loosely coiled into a short, thick chromatin fiber and estimated to contain about 3 kb DNA.

Intercalary heterochromatin of Drosophila as a potential model for human fragile sites.

We summarize our proposal that "intercalary heterochromatin" of Drosophila is a useful model for human fragile sites. Comparison with Drosophila site 11A suggests that the normal allele of fragile site Xq27 is a meiotic pairing site.

Three euchromatic DNA sequences under-replicated in polytene chromosomes of Drosophila are localized in constrictions and ectopic fibers.

We examined three regions of under-represented euchromatic DNA sequences (histone, Ubx, and 11 A), for their possible correlation with euchromatic constrictions in polytene chromosomes of Drosophila melanogaster. Cloned sequences were hybridized to filters and to chromosomes prepared for light microscopy. Under-represented sequences hybridized to DNA within constrictions and in ectopic fibers. In contrast, adjacent sequences that were fully endoreplicated in the Ubx and 11 A regions in polytene cells hybridized to sites just adjacent to their respective constrictions. For one region (Ubx), sequences under-represented in salivary gland cells were fully endoreplicated in fat body cells. For this particular region, the morphology of the polytene chromosomes differs between these two cell types in that the specific constriction is absent at this region in fat body polytene chromosomes, thus strengthening the correlation between under-representation and chromosome constrictions. Although all three sequences are in regions that have been classified by others as "intercalary heterochromatin," we detect no common functional or sequence organizational feature for these examples of under-represented DNA. We suggest that the lower efficiencies of the replication origins, or special regions of termination at these sites, are the primary cause of the under-replication, and that this under-replication is sufficient to confer the properties of intercalary heterochromatin.

Visualization of the formation and transport of a specific hnRNP particle.

The growth and maturation of the transcription products on the Balbiani ring (BR) genes in Chironomus tentans has been characterized by electron microscopy. The BR transcript is packed into a series of well defined ribonucleoprotein structures of increasing complexity: a 10 nm fiber, a 19 nm fiber, a 26 nm fiber, and a 50 nm granule. The basic 10 nm element was revealed in Miller spreads. The in situ structure of the transcription products and RNA compaction estimates suggested that the 10 nm fiber is packed into the 19 nm fiber as a tight coil. The transition of the 19 nm fiber into the 26 nm fiber is accompanied by a major change of the basic 10 nm fold into a noncoiled structure. Finally, the 26 nm fiber makes a one and one-third left-handed turn forming the final product, the BR granule. Upon translocation through the nuclear pore the BR granule becomes rod-shaped, which most likely corresponds to a relaxation of the highest-order structure into a straight 26 nm fiber.

Visualization of active 75 S RNA genes in the Balbiani rings of Chironomus tentans.

The active 75 S RNA genes in the Balbiani rings of Chironomus tentans were investigated by two complementing electron microscopy procedures: spreading of isolated chromosomes according to Miller and serial sectioning of Balbiani rings. The Miller spreads show that the transcription products, the ribonucleoprotein (RNP) fibers, increase gradually in length along the 75 S RNA genes. The morphology of the active genes in situ suggests the following sequence of events: during transcription a 20 nm RNP fiber is formed which is oriented roughly perpendicular to the chromosomal axis. When an RNP fiber corresponding to one quarter of the gene has been generated, packaging of the fiber into a dense globular structure begins at the free end of the RNP fiber. The processes of transcription and packaging go on in parallel in such a way that the length of the 20 nm fiber is kept constant, while the globular part is steadily increasing in size. When transcription is completed, a globular product is released, 50 nm in diameter. A thin (5 nm) chromosomal axis as well as a low DNA compaction (3.6) imply that the DNA in the chromosome fiber of an active 75 S RNA gene is more extended than the DNA in a nucleofilament. On the basis of our electron microscopy data and other information available on the Balbiani ring genes, we discuss the activation of the 75 S RNA genes in terms of a two-step process.

Characterization of active transcription units in Balbiani rings of Chironomus tentans.

Specific active transcription units on chromosome IV in the salivary glands of Chironomus tentans have been visualized by the Miller spreading technique and in situ by conventional electron microscopy. These units are likely to be located in the two most conspicuous puffs on chromosome IV, Balbiani ring 1 (BR 1) and Balbiani ring 2 (BR 2). The transcription units in these Balbiani rings generate 75S RNA molecules constituting putative messenger RNA species for the predominant cellular product, the salivary polypeptides. Solitary active transcription units with a mean length of 7.7 micron were observed most frequently. The lateral ribonucleoprotein (RNP) fibers of each unit formed a single length gradient. The number of fibers per unit was 123 (+/- 24), or about 16 growing RNP fibers per micron of chromosome fiber. The considerable variation in the number of RNP fibers per unit suggests that transcription can be modulated at the level of the individual gene. The modulation is probably achieved via the initiation event and/or via an early pretermination step, but a change in the elongation rate could not be excluded. The number of polymerases starting to traverse the whole gene was estimated to be six per min and transcription unit, and the rate of RNA chain elongation was calculated to be 31 nucleotides per second at 18 degrees C. The properties of the chromosome fiber within the active 75S RNA units and also within their vicinity were studied in the Miller spreads. The inactive chromosome fiber exhibited a uniform beaded conformation, while the active fiber was sparsely and irregularly beaded. Furthermore, the chromosome fiber was more extended in the active 75S RNA unit than in inactive regions (DNA packing ratios of 1.6 and 1.9, respectively). By comparing the properties of the active 75S RNA gene with those of active genes in other systems, it was inferred that the loss of beads and the extension of the fiber in the active unit is probably directly related to the level of transcriptive activity. Finally, a smooth nonbeaded segment of 0.18 micron in length was found to precede the RNP fiber gradient. This segment may have a role in the process of transcriptional regulation. On the basis of comparison with the active transcription units in spread preparations. It was possible to identify active units in the Balbiani rings in sectioned material using conventional electron microscopy. In both BR 1 and BR 2 an active unit appeared as a loop, consisting of a fiber axis and having RNP granules attached to the loop axis by stalks. The growing RNP fibers therefore seem to be organized into granular structures during the transcription process, and the final products in BR 1 and BR2 are granules, 500 A in diameter, each containing a 75S RNA molecule.

The 75 S RNA transcription unit in Balbiani ring 2 and its relation to chromosome structure.

A defined transcription unit in the Balbiani ring 2 (BR 2) region of chromosome IV in the salivary glands of Chironomus tentans has been characterized on the basis of analysis of the corresponding primary transcript, 75S RNA, and its functional significance. The available information on the transcription unit and its relations to chromosome structure can be summarized in the following way: 1. The size of the 75S RNA transcription unit in BR 2 is on the order of 30 000 base pairs. 2. The unit is likely to contain a long coding segment (at least 6000 base pairs), probably corresponding to information for salivary polypeptides. 3. The sequences are distributed in more than one chromomere (probably in 3--5 chromomeres). Further studies are needed before it can be stated whether or not there is a simple one-to-one relation between chromomeres and transcription units in the BR 2 region.

The size of poly(A)-containing RNAs in Drosophila melanogaster embryos.

The size range of poly(A)-containing RNA from Drosophila melanogaster embryos has been estimated by hybridization with 3H-labeled poly(U) and subsequent fractionation on sucrose gradients. The median size of nuclear poly(A)-containing RNA is about 30 S (6000 nucleotides), and the median size of cytoplasmic poly(A)-containing RNA is about 17 S (1800 nucleotides). The relationship of these sizes to messenger RNA needed to code for protein and to the length of DNA contained in a chromomere is discussed.