Age effects on intracortical microstructure and cortical thickness in adult saddle-back tamarins

Intracortical bone remodeling and cortical thickness data were collected from middiaphyseal thin sections of the humerus, ulna and tibia for 47 specimens ofSaguinus fuscicollis. Individuals were classified into age cohorts: Young Adult, Mid-adult I, Mid-adult II and Old Adult. Correlation analyses revealed significant relationships between age cohort and the number of osteons for the humerus and ulna, and between age cohort and non-Haversian canals for each of the three long bone elements. Significant relationships between age cohort and dimension of cortex size suggested an age-related pattern of cortical shifting in the ulna and older-age bone loss in the tibia.     

Colchicine inhibits plasmodium formation and disrupts pathways of sporopollenin secretion in the anther tapetum ofTradescantia virginiana L.

Summary During an earlier investigation, microtubules were observed at the periphery of invasion processes in the developing syncytial tapetum ofTradescantia virginiana L. They were also associated with membranous sacs that accumulate adjacent to tetrads, with putative fusion sites where the tapetal plasmodium is initiated, and, in postmeiotic stages, with the perispore membrane that encloses the developing spore cells. Colchicine was administered to developing flower buds to investigate the roles of these microtubules. The results indicate that microtubules neither initiate nor guide the tapetal invasion of the loculus. The treatments, however, resulted in absence of cell coat from invasion processes and prevention of cell fusion. They also inhibited polarized migration of membrane sacs and removed the associated microtubules. The development of an organized secretory apparatus at the perispore membrane was disrupted, with subsequent disordered deposition of sporopollenin in the extracellular spaces of the partially-fused plasmodium. The results suggest that microtubules participate in the formation and internal spatial organization of the tapetal plasmodium, and establishment of a secretory surface that normally produces sporopollenin at the tapetum-microspore interface.     

Phagocytosis of different matrix components by different cell types at bone-forming sites in cultured mouse calvariae

Summary Sites of bone formation on fragments of parietal bone of fetal-mice cultured for 10 days were examined by electron microscopy after addition of either ruthenium red or ferrocyanide to the postfixation fluid. Osteoclasts, osteoblast-like cells, and macrophages were the principal active cells at these formation sites. The mononuclear cells (osteoblast-like cells and macrophages) in the osteoid tissue showed evidence of having incorporated elements of calcified tissue. Osteoblast-like cells had phagocytized collagen fibrils and calcified bone matrix. This occurred more frequently in the calcifying area. Mononuclear macrophages showed not only phagocytosis and digestion of cellular debris and bone spicules in the osteoid, but also active incorporation of calcified bone matrix that had been detached from its surroundings by its pseudopod-like projections from long cytoplasmic processes. Collagen fibrils were seldom observed within the macrophages. These observations suggest that in our culture system osteoblast-like cells and macrophages at bone formation sites have a phagocytic role in bone remodeling.This study was supported in part by a grant from the Ministry of Education. Science and Culture of Japan (No. 59771321)     

海城小孤山的骨制品和装饰品

小孤山的骨制品包括鱼叉、标枪头和骨针,它们在工艺上与欧洲旧石器时代晚期、尤其是马格德林文化的制品十分相似。小孤山的骨针和山顶洞的骨针相似,但工艺水平稍高。小孤山的装饰品包括穿孔牙齿和蚌壳,它们与山顶洞的同类制品基本一致,但穿孔技术稍高。小孤山含上述制品的堆积物的时代为晚更新世,年代测定的初步结果为距今40,000—20,000年左右。     

Skeletal growth in a medieval population from Sudanese Nubia

Long bone growth is analyzed for 180 children from a Medieval population at Kulubnarti in Sudanese Nubia (550–1450 A.D.). A regional interpopulation comparison is made with growth data from Wadi Halfa in Lower Nubia, and an intrapopulation analysis is undertaken to assess diachronic changes in growth at Kulubnarti. Changes in growth patterns are interpreted in the context of mortality and morbidity profiles and relationships between the three variables are discussed. It is suggested that changes in the sociopolitical environment may have been responsible in part for altering levels of biological stress and impacting growth.     

L'os cellulaire d'un Poisson Téléostéen ≪Anguilla anguilla L.≫

Résumé Le squelette vertébral de l'Anguille est formé d'os cellulaire: lamellaire compact ou spongieux, suivant les différentes parties de la vertèbre. Nous avons pu y mettre en évidence, chez des animaux physiologiquement normaux, les trois catégories de cellules caractéristiques de l'os des vertébrés supérieurs: ostéoblastes, osteocytes, ostéoclastes. Elles ont les mêmes fonctions que chez ces derniers, les ostéoblastes procèdent à l'élaboration du tissu osseux, alors que les ostéoclastes le détruisent; à cette résorption ostéoclastique s'ajoute une lyse périostéocytaire ou résorption périlacunaire. Les osteocytes, dans ce tissu, nous paraissent être des éléments actifs; néanmoins, leur nombre (par unité de surface) est très inférieur à celui trouvé chez les Mammifères. Le remaniement osseux résultant du jeu de l'apposition et de la résorption est important et comparable à celui existant chez l'Homme.L'os de l'Anguille est, à de nombreux points de vue, très voisin de celui des Mammifères; les Poissons n'ont pas de parathyroïdes en tant que telles mais ils sont pourvus d'autres glandes vraisemblablement impliquées dans la régulation phosphocalcique: corps ultimobranchial et corpuscules de Stannius. Notre but sera donc d'essayer de déterminer comment est réglé le métabolisme de l'os cellulaire des Téléostéens.

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Histologic study on teleost cellular bone I.

Summary The vertebral skeleton of the eel consists of cellular bone, either lamellar or spongy, depending on different parts of the vertebra. In this osseous tissue we have found, in physiologically normal animals, three categories of caracteristic cells of the bone of higher vertebrates: osteoblasts, osteocytes, osteoclasts. They have the same functions as in higher vertebrates, osteoblasts form the bone while the osteoclasts which are to be found in Howship's lacunae destroy it. In addition, a perilacunar type of bone resorption or osteocytic osteolysis can be observed. In this bone, osteocytes seem to be active cells, but the concentration of osteocytes is decidedly lower than that to be found in Mammals.The osseous remodeling produced by apposition and resorption is of the same importance as in human bone. The bone of the eel, in many ways, closely resembles that of mammals. Teleost fish do not have parathyroid glands, but their phosphocalcic regulation seems to be facilitated by the action of two endocrine glands: Ultimobranchial body and the corpuscles of Stannius.The regulation of the cellular bone metabolism of Teleostean fishes is discussed.

Nous remercions Monsieur le Professeur Baud qui nous accueille dans son laboratoire à Genève et qui nous prodigue ses conseils.     

Single chloride channels in endosomal vesicle preparations from rat kidney cortex

Summary Endocytotic vesicles from rat kidney cortex, isolated by differential centrifugation and enriched on a Percoll gradient, contain both an electrogenic H⁺ translocation system and a conductive chloride pathway. Using the dehydration/rehydration method, we fused vesicles of enriched endosomal vesicle preparations and thereby made them accessible to the patch-clamp technique. In the fused vesicles, we observed Cl^– channels with a single-channel conductance of 73±2 pS in symmetrical 140mm KCl solution (n=25). The current-voltage relationship was linear in the range of –60 to +80 mV, but channel kinetic properties dependended on the clamp potential. At positive potentials, two sublevels of conductance were discernible and the mean open time of the channel was 10–15 msec. At negative voltages, only one substate could be resolved and the mean open time decreased to 2–6 msec. Clamp voltages more negative than –50 mV caused reversible channel inactivation. The channel was selective for anions over cations. Ion substitution experiments revealed an anion permeability sequence of Cl^–=Br^–=I^–>SO ₄ ^2– F^–. Gluconate, methanesulfonate and cyclamate were impermeable. The anion channel blockers 4,4-diisothiocyanatostilbene-2,2-disulfonic acid (DIDS, 1.0mm) and 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB, 0.1mm) totally inhibited channel activity. Comparisons with data obtained from radiolabeled Cl^–-flux measurements and studies on the H⁺ pump activity in endocytotic vesicle suspensions suggest that the channel described here is involved in maintenance of electroneutrality during ATP-driven H⁺ uptake into the endosomes.     

Fusion of liposomes and rat brain microsomes examined by two assays

Summary Liposomes are prepared from rat brain microsomal lipid and loaded with either Tb³⁺ or dipicolinic acid (DPA) to test fusion with the Tb-DPA assay. They are also loaded with octadecyl Rhodamine B chloride (R₁₈) to test fusion with the R₁₈ assay. The addition of either Ca²⁺ or Mg²⁺ to loaded liposomes develops fluorescence with both assays. The fluorescence elicited by Mg²⁺ is similar to that elicited by Ca²⁺ if assessed with R₁₈, but much higher if determined by Tb-DPA. The Ca²⁺-dependent fluorescence of the Tb-DPA complex is not suppressed by the addition of EDTA, and therefore it is internal to vesicles. The contrary is true for the Mg²⁺-dependent fluorescence. Rat brain microsomes can be disrupted by adding octylgucoside and reconstituted by removing it by dialysis. We use this procedure to load microsomes with DPA. This allows the use of the Tb-DPA assay for testing the fusion of rat brain microsomes. Reconstituted microsomes fuse with liposomes. This fusion has characteristics similar to those of liposome-liposome fusion. However, no microsome-microsome fusion could be detected with either method. The two methods give different results, owing to the chemical properties of the assays. Indeed Tb-DPA implies the retention of vesicle content, whereas this is not required by the R₁₈ assay.     

Fusion of cultured dog kidney (MDCK) cells: I. Technique,fate of plasma membranes and of cell nuclei

Summary The evaluation of the intracellular signal train and its regulatory function in controlling transepithelial transport with electrophysiological methods often requires intracellular measurements with microelectrodes. However, multiple impalements in epithelial cells are hampered by the small size of the cells. In an attempt to avoid these problems we fused cells of an established cell line, Madin Darby canine kidney cells, originally derived from dog kidney, to giant cells by applying a modified polyethylene glycol method. During trypsin-induced detachment from the ground of the petri dish, individual cells grown in a monolayer incorporate volume and mainly lose basolateral plasma membrane by extrusion. By isovolumetric cell-to-cell fusion, spherical giant cells are formed within 2 hr. During this process a major part of the individual cell plasma membranes is internalized. Over three weeks following cell plasma membrane fusion degradation of single cell nuclei and cell nuclear fusion occurs. We conclude that this experimental approach opens the possibility to investigate ion transport of epithelia in culture by somatic cell genetic techniques.     

Molecular evolution of enzyme structure: Construction of a hybrid hamster/Escherichia coli aspartate transcarbamoylase

Summary Aspartate transcarbamoylase (ATCase, EC 2.1.3.2) is the first unique enzyme common to de novo pyrimidine biosynthesis and is involved in a variety of structural patterns in different organisms. InEscherichia coli, ATCase is a functionally independent, oligomeric enzyme; in hamster, it is part of a trifunctional protein complex, designated CAD, that includes the preceding and subsequent enzymes of the biosynthetic pathway (carbamoyl phosphate synthetase and dihydroorotase). The complete complementary DNA (cDNA) nucleotide sequence of the ATCase-encoding portion of the hamster CAD gene is reported here. A comparison of the deduced amino acid sequences of the hamster andE. coli catalytic peptides revealed an overall 44% amino acid similarity, substantial conservation of predicted secondary structure, and complete conservation of all the amino acids implicated in the active site of theE. coli enzyme. These observations led to the construction of a functional hybrid ATCase formed by intragenic fusion based on the known tertiary structure of the bacterial enzyme. In this fusion, the amino terminal half (the “polar domain”) of the fusion protein was provided by a hamster ATCase cDNA subclone, and the carboxyl terminal portion (the “equatorial domain”) was derived from a clonedpyrBI operon ofE. coli K-12. The recombinant plasmid bearing the hybrid ATCase was shown to satisfy growth requirements of transformedE. coli pyrB ⁻ cells. The functionality of thisE. coli-hamster hybrid enzyme confirms conservation of essential structure-function relationships between evolutionarily distant and structurally divergent ATCases.