Corneal collagen fibril structure in three dimensions: Structural insights into fibril assembly, mechanical properties, and tissue organization

The ability of the cornea to transmit light while being mechanically resilient is directly attributable to the formation of an extracellular matrix containing orthogonal sheets of collagen fibrils. The detailed structure of the fibrils and how this structure underpins the mechanical properties and organization of the cornea is understood poorly. In this study, we used automated electron tomography to study the three-dimensional organization of molecules in corneal collagen fibrils. The reconstructions show that the collagen molecules in the 36-nm diameter collagen fibrils are organized into microfibrils (≈4-nm diameter) that are tilted by ≈15° to the fibril long axis in a right-handed helix. An unexpected finding was that the microfibrils exhibit a constant-tilt angle independent of radial position within the fibril. This feature suggests that microfibrils in concentric layers are not always parallel to each other and cannot retain the same neighbors between layers. Analysis of the lateral structure shows that the microfibrils exhibit regions of order and disorder within the 67-nm axial repeat of collagen fibrils. Furthermore, the microfibrils are ordered at three specific regions of the axial repeat of collagen fibrils that correspond to the N- and C-telopeptides and the d-band of the gap zone. The reconstructions also show macromolecules binding to the fibril surface at sites that correspond precisely to where the microfibrils are most orderly.

Spatial and Temporal Analyses of Expansion and Cell Cycle in Sunflower Leaves1 : A Common Pattern of Development for All Zones of a Leaf and Different Leaves of a Plant

We have investigated the spatial distributions of expansion and cell cycle in sunflower (Helianthus annuus L.) leaves located at two positions on the stem, from leaf initiation to the end of expansion. Relative expansion rate (RER) was analyzed by following the deformation of a grid drawn on the lamina; relative division rate (RDR) and flow-cytometry data were obtained in four zones perpendicular to the midrib. Calculations for determining in situ durations of the cell cycle and of S-G2-M in the epidermis are proposed. Area and cell number of a given leaf zone increased exponentially during the first two-thirds of the development duration. RER and RDR were constant and similar in all zones of a leaf and in all studied leaves during this period. Reduction in RER occurred afterward with a tip-to-base gradient and lagged behind that of RDR by 4 to 5 d in all zones. After a long period of constancy, cell-cycle duration increased rapidly and simultaneously within a leaf zone, with cells blocked in the G0-G1 phase of the cycle. Cells that began their cycle after the end of the period with exponential increase in cell number could not finish it, suggesting that they abruptly lost their competence to cross a critical step of the cycle. Differences in area and in cell number among zones of a leaf and among leaves of a plant essentially depended on the timing of two events, cessation of exponential expansion and of exponential division.

The physical and genomic organization of microsatellites in sugar beet.

Microsatellites, tandem arrays of short (2-5 bp) nucleotide motifs, are present in high numbers in most eukaryotic genomes. We have characterized the physical distribution of microsatellites on chromosomes of sugar beet (Beta vulgaris L.). Each microsatellite sequence shows a characteristic genomic distribution and motif-dependent dispersion, with site-specific amplification on one to seven pairs of centromeres or intercalary chromosomal regions and weaker, dispersed hybridization along chromosomes. Exclusion of some microsatellites from 18S-5.8S-25S rRNA gene sites, centromeres, and intercalary sites was observed. In-gel and in situ hybridization patterns are correlated, with highly repeated restriction fragments indicating major centromeric sites of microsatellite arrays. The results have implications for genome evolution and the suitability of particular microsatellite markers for genetic mapping and genome analysis.

Glutamate receptor blockade at cortical synapses disrupts development of thalamocortical and columnar organization in somatosensory cortex.

The segregation of thalamocortical inputs into eye-specific stripes in the developing cat or monkey visual cortex is prevented by manipulations that perturb or abolish neural activity in the visual pathway. Such findings show that proper development of the functional organization of visual cortex is dependent on normal patterns of neural activity. The generalisation of this conclusion to other sensory cortices has been questioned by findings that the segregation of thalamocortical afferents into a somatotopic barrel pattern in developing rodent primary somatosensory cortex (S1) is not prevented by activity blockade. We show that a temporary block of N-methyl-D-aspartate (NMDA) and non-NMDA glutamate receptors in rat S1 during the critical period for barrel development disrupts the topographic refinement of thalamocortical connectivity and columnar organization. These effects are evident well after the blockade is ineffective and thus may be permanent. Our findings show that neural activity and specifically the activation of postsynaptic cortical neurons has a prominent role in establishing the primary sensory map in S1, as well as the topographic organization of higher order synaptic connections.

Visualization of glucocorticoid receptor translocation and intranuclear organization in living cells with a green fluorescent protein chimera.

A highly fluorescent mutant form of the green fluorescent protein (GFP) has been fused to the rat glucocorticoid receptor (GR). When GFP-GR is expressed in living mouse cells, it is competent for normal transactivation of the GR-responsive mouse mammary tumor virus promoter. The unliganded GFP-GR resides in the cytoplasm and translocates to the nucleus in a hormone-dependent manner with ligand specificity similar to that of the native GR receptor. Due to the resistance of the mutant GFP to photobleaching, the translocation process can be studied by time-lapse video microscopy. Confocal laser scanning microscopy showed nuclear accumulation in a discrete series of foci, excluding nucleoli. Complete receptor translocation is induced with RU486 (a ligand with little agonist activity), although concentration into nuclear foci is not observed. This reproducible pattern of transactivation-competent GR reveals a previously undescribed intranuclear architecture of GR target sites.

The genomic and physical organization of Ty1-copia-like sequences as a component of large genomes in Pinus elliottii var. elliottii and other gymnosperms.

A DNA sequence, TPE1, representing the internal domain of a Ty1-copia retroelement, was isolated from genomic DNA of Pinus elliottii Engelm. var. elliottii (slash pine). Genomic Southern analysis showed that this sequence, carrying partial reverse transcriptase and integrase gene sequences, is highly amplified within the genome of slash pine and part of a dispersed element >4.8 kbp. Fluorescent in situ hybridization to metaphase chromosomes shows that the element is relatively uniformly dispersed over all 12 chromosome pairs and is highly abundant in the genome. It is largely excluded from centromeric regions and intercalary chromosomal sites representing the 18S-5.8S-25S rRNA genes. Southern hybridization with specific DNA probes for the reverse transcriptase gene shows that TPE1 represents a large subgroup of heterogeneous Ty1-copia retrotransposons in Pinus species. Because no TPE1 transcription could be detected, it is most likely an inactive element--at least in needle tissue. Further evidence for inactivity was found in recombinant reverse transcriptase and integrase sequences. The distribution of TPE1 within different gymnosperms that contain Ty1-copia group retrotransposons, as shown by a PCR assay, was investigated by Southern hybridization. The TPE1 family is highly amplified and conserved in all Pinus species analyzed, showing a similar genomic organization in the three- and five-needle pine species investigated. It is also present in spruce, bald cypress (swamp cypress), and in gingko but in fewer copies and a different genomic organization.

O nado de atletas de águas abertas: características do desempenho e da organização temporal das braçadas

A natação de águas abertas tem registrado aumento no número de competições e participantes em todo mundo. Acompanhando esta tendência têm sido desenvolvidos estudos para identificar as características físicas e as respostas fisiológicas dos atletas neste tipo de prova. Entretanto, são escassos estudos ao nível de análise comportamental, principalmente, em condições reais de distância e meio ambiente (mar). Foi objetivo deste estudo investigar as características de desempenho e da organização temporal das braçadas de nadadores de águas abertas. Mais especificamente, conhecer quais recursos os atletas de águas abertas lançam mão para atingir sua meta de vencer um percurso no mar no menor tempo possível. A amostra foi constituída por 23 atletas, com média de idade de 26,4(±3,2) anos. A tarefa foi nadar um trajeto de 1500 metros em forma de um circuito em mar aberto. Para a captação das variáveis relacionadas ao desempenho utilizou-se um GPS (Garmin modelo Fênix 3) e um cronômetro (FINIS modelo Accusplit Eagle AX602). O registro das imagens para captação dos dados relacionados à descrição da organização temporal das braçadas ocorreu em três pontos do trajeto: início (I) - 20 a 40 metros, meio (M) - 800 a 820 metros e final (F) - 1450 a 1470 metros. Foi utilizada uma filmadora (Nikon Coolpix S5300) afixada à embarcação. O software Kinovea 8.20 permitiu a análise quadro a quadro das braçadas. Foram consideradas variáveis dependentes relacionadas ao desempenho (tempo, velocidade e distância total percorrida, bem como, a frequência de braçadas em cada um dos três pontos do trajeto); aos aspectos variantes das braçadas (tempo total do ciclo, das braçadas, das fases aérea e aquática) e aos aspectos invariantes das braçadas (timing relativo das fases aérea e aquática e sua variabilidade). A análise de variância de medidas repetidas foi usada para comparar os três momentos da tarefa (I, M e F) para todas as variáveis, e a correlação de Pearson para analisar a magnitude das relações entre as variáveis de desempenho, enquanto o teste t de Student para medidas pareadas foi utilizado para comparar as possíveis diferenças entre os braços direito e esquerdo para cada um dos momentos e determinou-se como significância estatística α≤=0,05. Em relação ao desempenho, os resultados indicaram que os nadadores fizeram uso de frequência de braçada (Fb) diferente para os três momentos, sendo maior no I quando comparada ao M e F, e no M, menor que em F; estas mudanças foram acompanhadas por ajustes nos aspectos variantes como o tempo total do ciclo, das braçadas e das fases aérea e aquática. Ainda, nos três momentos os nadadores apresentaram simetria temporal entre as braçadas dos dois braços, apesar de as diferenças serem evidenciadas entre as fases das braçadas quando comparados os braços. Com relação aos aspectos invariantes detectou-se mudança do padrão de I para M e F da tarefa, sendo que em M e F os atletas utilizaram a mesma estrutura temporal. Quanto à variabilidade dos aspectos variantes e invariantes para as braçadas e as fases das braçadas, observou-se diminuição da magnitude ao longo da tarefa sendo que o braço esquerdo apresentou nos três momentos maior variabilidade que o direito. Assim, diante dos resultados, concluiu-se que os recursos utilizados por nadadores habilidosos para nadar em ambiente pouco estável, em condições reais de distância e meio ambiente (mar) compreendem a alteração do desempenho (Fb) associado a ajustes nos aspectos variantes, concomitantemente à alteração dos aspectos invariantes das braçadas, em função do momento da tarefa

The episcopal complex of Eio-El Tolmo de Minateda (Hellín, Albacete, Spain). Architecture and spatial organization. 7th to 8th centuries AD

The episcopal complex of Eio, located in El Tolmo de Minateda, was built between the end of the 6th century and the beginning of the 7th century, possibly as a political decision taken by the ecclesiastical authority in the capital of the Visigothic kingdom (Toletum). With the comprehensive study of the whole complex presented below (construction cycles, furniture, decoration and location of spaces), we can interpret the function of each space in the basilica and the domus episcopi, the liturgical and general movement routes, the existence of some hierarchical environments, and specify the chronological development of the buildings. After the Arab-Berber conquest of Hispania in the early 8th century, the whole complex will experience a series of transformations that will convert the religious and monumental public area into a private, residential and industrial Islamic quarter.

Taylor's sacred minstrel, or, American church music book : a new collection of psalm and hymn tunes adapted to the various metres now in use : together with anthems, sentences, chants and other pieces : for the use of choirs, congregations, singing schools, musical societies, social sacred music circles, and private devotion ... /

Mode of access: Internet.

Latin America timber trends and prospects [by] Economic Commission for Latin America [and] Food and Agriculture Organization of the United Nations.

"Errata": 1 leaf inserted.

The Army Service Corps of the British Army, and, the organization of the transport and transportation at the front in France : lectures delivered before the officers of the Quartermaster Corps and Quartermaster Reserve Corps at Washington, D.C., on May 2 and May 9, 1917 respectively /

Mode of access: Internet.

Nineteen sonatas for the piano. [music] : [English and Spanish] /

Mode of access: Internet.