Innervation of human omental arteries and veins and vasomotor response to noradrenaline, neuropeptide Y, substance P and vasoactive intestinal peptide

Human omental arteries and veins are supplied with nerve fibers containing noradrenaline (NA) and neuropeptide Y (NPY); these two agents probably co-exist in perivascular sympathetic nerve fibers. Substance P (SP)- or vasoactive intestinal peptide (VIP)-containing fibers could not be detected. In studies on isolated omental vessels NA produced constriction. The results of blockade experiments suggest that human omental arteries are equipped predominantly with alpha 1-adrenoceptors and omental veins with a mixture of alpha 1- and alpha 2-adrenoceptors. NPY at a concentration of 10(-7) M or higher had a weak contractile effect on veins and virtually no effect on arteries. NPY at a concentration of 3 X 10(-8) M shifted the NA concentration response curve to the left in arteries (pD2 = 5.8 for NA versus 6.6. for NA in the presence of NPY; P less than 0.001) but not in veins. Both SP and VIP relaxed arteries precontracted with NA or prostaglandin F2 alpha (PGF2 alpha). The potency of SP as a relaxant agent was similar in arteries and veins; the effect of VIP was elicited at lower concentrations in veins than in arteries.     

Development of the orangutan permanent dentition: Assessing patterns and variation in tooth development

This study examines dental formation and alveolar emergence in a large cross-sectional sample composed primarily of wild-reared orangutans (N = 89) in order to provide information on the development of the permanent dentition in this hominoid and to address questions of variation in individual tooth formation, between teeth and between individuals. All specimens have been radiographed in lateral aspect and stages of crown and root formation recorded for all teeth. The ranges of crown and root formation of I¹₁, C¹₁, P⁴₄, and M³₃ have been calculated relative to the stage of M₁¹ development within a specific tooth quadrant. Then, for each specimen, BMDP scatterplot and nonparametric statistics have been used to graph changes in stages of these teeth relative to M¹₁ stages and to examine relationships between pairs of upper and lower dental counterparts and between teeth of each jaw. Results indicate 1) high correlations between upper and lower tooth pairs and between many of the permanent teeth within individuals, 2) a relatively large range of variability in individual tooth development (multistage ranges relative to M¹₁), 3) greater variation in root development at emergence than earlier reports, and 4) evidence of variability within the sequence emergence pattern of the orangutan. © 1996 Wiley-Liss, Inc.     

Occurrence of Chaperonin 60 and Chaperonin 10 in primary and secondary bacterial symbionts of aphids: Implications for the evolution of an endosymbiotic system in aphids

Summary All aphids harbor symbiotrophic prokaryotes (primary symbionts) in a specialized-abdominal cell, the bacteriocyte. Chaperonin 60 (Cpn60, symbionin) and chaperonin 10 (Cpn10), which are high and low molecular weight heatshock proteins, were sought in tissues of more than 60 aphid species. The endosymbionts were compared immunologically and histologically. It was demonstrated that (1) there are two types of aphids in terms of the endosymbiotic system: some with only primary symbionts and others with, in addition, secondary symbionts; (2) the primary symbionts of various aphids are quite similar in morphology whereas the secondary symbionts vary; and (3) irrespective of the aphid species, Cpn60 is abundant in both the primary and secondary symbionts, while Cpn10 is abundant in the secondary symbionts but present in small amounts in the primary ones. Based on these results, we suggest that the primary symbionts have been derived from a prokaryote that was acquired by the common ancestor of aphids whereas the secondary symbionts have been acquired by various aphids independently after divergence of the aphid species. In addition, we point out the possibility that the prokaryotes under intracellular conditions have been subject to some common evolutionary pressures, and as a result, have come to resemble cell organelles.     

Comparison of the distribution patterns of BK polyomavirus lineages among China, Korea and Japan: Implications for human migrations in northeast Asia

BKV is widespread among humans, infecting children asymptomatically and then persisting in renal tissue. Based on the serological or phylogenetic method, BKV isolates worldwide are classified into four subtypes (I–IV), with subtypes I and IV further divided into several genetically-distinct subgroups. Since, similarly to JCV, a close relationship exists between BKV lineages and human populations, BKV should be useful as a marker to trace human migrations. To elucidate ancient human migrations in northeast Asia, urine samples were collected from immunocompetent elderly patients in Shanghai, China; Anyang, South Korea; and various locations in Japan. Partial and complete BKV genomes from these samples were amplified and sequenced using PCR, and the determined sequences were classified into subtypes and subgroups by phylogenetic and SNP analyses. In addition, based on an SNP analysis, the major subtype I subgroup (I/c) was classified into two subdivisions, I/c/Ch and I/c/KJ. The distribution patterns of BKV subgroups and subdivisions among the three regions were compared. Some aspects of the subgroup and subdivision distribution were more similar between Korea and Japan, but others were more similar between China and Korea or between China and Japan. Based on these findings, we inferred various northeast Asian migrations. Most of the JCV-based inferences of northeastern Asian migrations were consistent with those based on BKV, but the previously suggested migration route from the Asian continent to the Japanese archipelago seemed to need revision.     

Anomalous branching patterns in some fossilFilicales: Implications in the evolution of the megaphyll and the lateral branch,habit and growth pattern

From middle Carboniferous coal balls leaves and stems of structurally preserved Filicalean ferns (Psalixochlaenaceae, Botryopteridaceae) are described that deviate from the normal pattern of branching. From this, interpretations are made as to the origin and relationship of axillary branching and dichotomy in this line of ferns. Their megaphylls apparently have orginated from a three-dimensional dichotomous branching system. This is inferred from the widespread occurrence of short, leafy epiphyllous buds on their fronds and is related to their habit where the leaf often plays the dominant role and effects vegetative propagation.     

Tarsal and tarsometatarsal coalitions from Mound C (Ocmulgee Macon Plateau site,Georgia): Implications for understanding the patterns,origins, and antiquity of pedal coalitions in Native American populations

Tarsal and tarsometatarsal coalitions are malsegmentation errors that result in incomplete division between two or more normally separate bones of the foot. Coalitions may be osseous, characterized by bony union, or non-osseous, in which the affected elements are united by fibrous tissue, cartilage, or some combination of both. Evidence indicates that non-osseous coalitions are frequently overlooked or misinterpreted in skeletal samples. The purpose of this study is to (1) report two non-osseous coalition cases (naviculocuneiform I, CF3-MT3) from the Ocmulgee Mound Site in Georgia, and (2) examine the occurrence of coalitions throughout the foot in Native American samples relative to other major populations. Evidence suggests that Native Americans exhibit a pattern of coalitions in the foot that differs from that recently documented for European and African samples. Native Americans display a relatively high rate of midfoot and forefoot coalitions, and known cases are both geographically and temporally diverse. This distribution, along with evidence of similar patterns in East Asian samples, suggests that the pattern of coalition seen in Native Americans has origins in Asiatic parent populations during the late Pleistocene. Individuals migrating to the New World with proximal midfoot coalitions are likely to have endured biomechanical stress during prolonged physical activity and walking, as frequently seen in modern clinical cases.     

Manipulation of environmental variables and the effect on the growth of Haliclona sp.: Implications for open-water aquaculture

We examined how the physical environment influences the growth and survival of an undescribed Haliclona species. To determine the influence that water movement, light and sediment had on the sponge, explants of Haliclona sp. (approximately 8 cm³ in size) were transplanted into manipulated microenvironments at Hamelin Bay on the west coast of Western Australia near Perth. The sponge is typically found under limestone ledges and appears to have distinct limits on the microenvironment in which it is found. A three-factor orthogonal design was used to manipulate levels of light, water flow and sedimentation. Each factor had two experimental levels, creating environments with high and low water movement, high and low light, and upward and downward orientations to control sediment levels. The survival of explants was high (100%). However, all explants showed a regression in weight. Explants transplanted on to the underside of horizontal surfaces (downward orientations) demonstrated significantly less weight loss (P = 0.023), which was attributed to lower sediment exposure. Light and water movement did not significantly influence the sponge's growth.     

Heartbeat control in the medicinal leech: A model system for understanding the origin,coordination, and modulation of rhythmic motor patterns

We have analyzed in detail the neuronal network that generates heartbeat in the leech. Reciprocally inhibitory pairs of heart interneurons form oscillators that pace the heartbeat rhythm. Other heart interneurons coordinate these oscillators. These coordinating interneurons, along with the oscillator interneurons, form an eight-cell timing oscillator network for heartbeat. Still other interneurons, along with the oscillator interneurons, inhibit heart motor neurons, sculpting their activity into rhythmic bursts. Critical switch interneurons interface between the oscillator interneurons and the other premotor interneurons to produce two alternating coordination states of the motor neurons. The periods of the oscillator interneurons are modulated by endogenous RFamide neuropeptides. We have explored the ionic currents and graded and spike-mediated synaptic transmission that promote oscillation in the oscillator interneurons and have incorporated these data into a conductance-based computer model. This model has been of considerable predictive value and has led to new insights into how reciprocally inhibitory neurons produce oscillation. We are now in a strong position to expand this model upward, to encompass the entire heartbeat network, horizontally, to elucidate the mechanisms of FMRFamide modulation, and downward, to incorporate cellular morphology. By studying the mechanisms of motor pattern formation in the leech, using modeling studies in conjunction with parallel physiological experiments, we can contribute to a deeper understanding of how rhythmic motor acts are generated, coordinated, modulated, and reconfigured at the level of networks, cells, ionic currents, and synapses. © 1995 John Wiley & Sons, Inc.     

Limb‐bone histology of temnospondyls: implications for understanding the diversification of palaeoecologies and patterns of locomotion of Permo‐Triassic tetrapods

The locomotion of early tetrapods has long been a subject of great interest in the evolutionary history of vertebrates. However, we still do not have a precise understanding of the evolutionary radiation of their locomotory strategies. We present here the first palaeohistological study based on theoretical biomechanical considerations among a highly diversified group of early tetrapods, the temnospondyls. Based on the quantification of microanatomical and histological parameters in the humerus and femur of nine genera, this multivariate analysis provides new insights concerning the adaptations of temnospondyls to their palaeoenvironments during the Early Permian, and clearly after the Permo‐Triassic crisis. This study therefore presents a methodology that, if based on a bigger sample, could contribute towards a characterization of the behaviour of species during great evolutionary events.     

The effect of inoculum size on the growth of cell and root cultures ofHyoscyamus muticus: Implications for reactor inoculation

Cell suspensions inoculated at low cell concentrations displayed a typical growth reduction, whereas root cultures displayed an improvement in growth. Specific growth rate ofHyoscyamus muticus cell suspensions decreased from 0.25 to 0.12 d⁻¹ as inoculum concentration was reduced from 4.0 to 0.02 g fresh weight per liter. In contrast, roots show an increase in growth rate from 0.24 to 0.43 d⁻¹. These contrasting growth patterns can be explained as the result of: a) the high specific surface area of cells as compared to roots and, b) the differentiated structure of roots. The dispersed nature of cell suspensions makes them more prone to leakage of key growth factors/cellular contents to medium. The results of this work indicate that cell cultures require substantially higher inoculum concentrations. In contrast, roots can be inoculated at very low concentrations. These facts imply that whereas seed vessels must be employed by cell suspensions, their use for root cultures is a compromise between an easier handling of an entwined root mass and the reduction of the contamination risk of large medium volumes.     

Effects of defined medium,fetal bovine serum,and human serum on growth and chemosensitivities of human breast cancer cells in primary culture: Inference for in vitro assays

Summary We compared the effects of defined medium, fetal bovine serum (FBS) and human serum (HuS) on the growth and responses to chemotherapeutic agents of human breast cancer cells in primary culture. Normal and tumor tissues were dissociated to small aggregates and single cells and seeded onto collagen-gel-coated wells in defined medium or medium supplemented with 5% FBS or 5% HuS. In all cases examined, defined medium and medium containing HuS were superior to medium containing FBS in supporting growth of both normal and tumor cell cultures. However, cultures in defined medium showed an initial cell loss. Cells from the same tumor cultured in different media varied in their responses to chemotherapeutic agents. In light of these results, medium supplemented with HuS, which promoted attachment of these cells in culture and stimulated their growth, should be the most appropriate nutrient environment for determining the effects of therapeutic agents on cells as it most closely resembles the in vivo situation. Because there were also variations in growth rates and chemosensitivities of tumor cells cultured in different human serum samples, we suggest that optimal conditions in which to culture these cells include the serum of the patient whose tumor is removed. This serum may provide host factors that influence cell growth and interact with exogenous factors. This work was supported by a grant from the National Cancer Institute of Canada and funds contributed by Mr. B. T. Wharton in memory of his wife, Nadia. J. T. Emerman is a research scholar of the National Cancer Institute of Canada.     

Morphological characterization and molecular phylogeny of Portunoidea Rafinesque, 1815 (Crustacea Brachyura): Implications for understanding evolution of swimming capacity and revision of the family-level classification

Cheliped construction, in particular the teeth pattern on chelae fingers is considered as most important character suit (along with burrowing/swimming apparatus) for the diagnosis of Portunoidea. Heterochelic and heterodontic chelipeds with the molariform tooth in the larger chela and multi-lobed serial teeth are presumably ancestral and most common pattern for the group. New material (mostly species of Thalamitinae Paulson, 1875, Lupocyclus Adamd and White, 1848 and Portunus Weber, 1795 sensu lato) have been combined with the existing sequences from the GenBank to produce molecular phylogenetic reconstructions based on the histone H3 gene fragment and a multi-gene tree (for smaller set of species) based on partial sequences of H3, D1 region of 28S gene and mitochondrial COI gene. These reconstructions have not provided necessary support to the monophyly of Portunoidea sensu lato but indicated the presence of several monophyletic lineages, i.e. Portunidae sensu stricto, Polybiidae + Thiidae + Carcinidae + Pirimelidae, Benthochascon + Geryonidae (to lesser extent), and Ovalipes. Monophyly of the Portunidae sensu stricto is supported by both the H3 and multigene trees and morphological evidence. Swimming capacity probably evolves as a result of parallel evolution in at least three different lineages of portunoids. A new version of the family level classification of Portunoidea and a key to their families are provided with the following taxa: Geryonidae (Geryoninae + Benthochasconinae subfam. nov.), Ovalipidae fam. nov., Brusiniidae Štev?i?, 1991, Thiidae, Pirimelidae, Carcinidae McLeay, 1838 (Carcininae + Portumninae Ortmann, 1893), Polybiidae Ortmann, 1893, and Portunidae Rafinesque, 1815 sensu stricto. The most radical change in the systematics of Portunidae sensu stricto is the final recognition of the polyphyly of Portunus sensu lato and the need for revalidization and re-diagnozing of several taxa that were synonymized by Stephenson and Campbell (1959) and Stephenson (1972) under Portunus. While some subfamilies of the Portunidae (Podophthalminae Dana, 1851, Thalamitinae, and Lupocyclinae Alcock, 1895) are well supported by molecular phylogenies and the presence of morphological synapomorphies, the others need re-assessment.     

Transforming growth factor-β in the early mouse embryo: Implications for the regulation of muscle formation and implantation

In a search for functions of transforming growth factor-β during early embryonic development we used two different experimental approaches. In the first we made use of embryonic stem (ES) cells. ES cells in culture differentiate to derivatives of all three germ layers and mimic some aspects of organogenesis when grown as aggregates in suspension to form embryoid bodies. Differentiation procedes further when the embryold bodies attach to suitable substrates. Muscle and neuronal cells are among the most readily identified cell types then formed. We examined the effect of all-trans retinoic acid (RA) and members of the transforming growth factor-β family(TGF-βl, TGF-β2) under these conditions in an assay where single aggregates formed in hanging microdrops in medium supplemented with serum depleted of lipophilic substances which would include retinoids. Endoderm-like cells formed under all conditions tested. RA at concentrations of 10⁸ M and 10⁷ M induced the formation of neurons but in the absence of RA or at concentrations up to 10?⁹ M, neurons were not observed. Instead, beating muscle formed in about one-third of the plated aggregates; this was greatly reduced when RA concentrations increased above 10?⁹ M. Immunofluorescent staining for muscle specific myosin showed that two muscle cell types could be distinguished: elongated, non-contractile myoblasts and mononucleate flat cells. The mononucleate flat cells appeared to correspond with rhythmically contracting muscle. The number of non-contractile myoblasts increased 3-fold over controls in the presence of 10?⁹ M RA. TGF-βs increased the number of contractile and non-contractile muscle cells by a factor 3 to 7 over controls, depending on the TGF-β isoform added and the muscle cell type formed. TGF-β2 also invariably increased the rate at which contracting muscle cells were first observed in replated aggregates. The stimulatory effect of TGF-βs on the formation of mononucleate flat cells was completely abrogated by RA at 10?⁹ M while the number of myoblasts under similar conditions was unchanged. These data suggest that a complex interplay between retinoids and TGF-β isoforms may be involved in regulation of differentiation in early myogenesis. In the second approach, neutralizing polyclonal rabbit antibodies specific for TGF-β2 were injected into the cavity of mouse blastocysts 3.5 days post coitum (pc). After 1 day in culture, embryos were transferred to pseudopregnant females. The number of decidua, embryos and resorptions were counted at day 8.5–9.5 pc. Control antibody injected embryos implanted with high efficiency (87%) compared with anti-TGF-β2 injected embryos which implanted with an efficiency of only 43%. If empty decidua (resorptions) were included, the overall recovery was 71% and 32% for control and experimental embryos, respectively. Embryos that were recovered showed no overt macroscopic abnormalities. These results together impiy functions for TGF-βs in implantation as well as in later development of the embryo. © 1993Wiley-Liss, Inc.     

Human blood-derived fibrin releasates: Composition and use for the culture of cell lines and human primary cells

We have evaluated the capacity of two human blood fractions to substitute for FBS as growth medium supplement for human and animal cell cultures. Non-anticoagulated blood from volunteer donors (N = 13) was centrifuged to isolate a supernatant serum (SS) and a platelet-rich fibrin (PRF) clot which was squeezed to extract the releasate (PRFR). Both materials were characterized for the content in PDGF-AB, TGF-β1, VEGF, bFGF, EGF, IGF, total protein, albumin, IgG, IgM IgA, fibrinogen, cholesterol, triglycerides, various chemistry analytes and hemoglobin. Cell growth promoting activity of pooled SS and PRFR at 1, 5, and 10% in growth medium was evaluated over 7 days using human (HEK293, MG-63) and animal (SIRC, 3T3) cell lines and two human primary cultures (gingival fibroblasts and periodontal ligaments). Viable cell count was compared to that in cultures in FBS free-medium and 10% FBS supplement. SS and PRFR at 1-10% stimulated cell growth significantly more than FBS-free medium and in a way similar to 10% FBS in all cultures apart from 3T3. These two human blood-derived fibrin releasates are equally efficient to substitute for FBS as supplement for cell cultures and could be useful for specialized applications in regenerative medicine, dentistry and oral implantology, or cell therapy.     

Morphological and biochemical characterization of mineralizing primary cultures of avian growth plate chondrocytes: Evidence for cellular processing of Ca2+ and Pi prior to matrix mineralization

Advances in the culture of mineralizing growth plate chondrocytes provided an opportunity to study endochondral calcification under controlled conditions. Here we report that these cultures synthesize large amounts of proteins characteristically associated with mineralization: type II and X collagens, sulfated proteoglycans, alkaline phosphatase, and the bone-related proteins, osteonectin and osteopontin. Certain chondrocytes appeared to accumulate large amounts of Ca²⁺ and Pi during the mineralization process: laser confocal imaging revealed high levels of intracellular Ca²⁺ in their periphery and X-ray microanalytical mapping revealed the presence of many Ca²⁺- and Pi-rich cell surface structures ranging from filamentous processes 0.14 ± 0.02 μm by 0.5–2.0 μm, to spherical globules 0.70 ± 0.27 μm in diameter. Removal of organic matter with alkaline sodium hypochlorite revealed numerous deposits of globular (0.77 ± 0.19 μm) mineral (calcospherites) in the lacunae around these cells. The size and spatial distribution of these mineral deposits closely corresponded to the Ca²⁺-rich cell surface blebs. The globular mineral progressively transformed into clusters of crystallites. Taken with earlier studies, these findings indicate that cellular uptake of Ca²⁺ and Pi leads to formation of complexes of amorphous calcium phosphate, membrane lipids, and proteins that are released as cell surface blebs analogous to matrix vesicles. These structures initiate development of crystalline mineral. Thus, the current findings support the concept that the peripheral intracellular accumulation of Ca²⁺ and Pi is directly involved in endochondral calcification.