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Terpenoids with a new skeleton and novel triterpenoids with anti-inflammatory effects from Garcinia subelliptica.

Three novel phloroglucinol derivatives, garcinielliptones F (1), H (3), and I (4), and two novel terpenoids, garcinielliptones G (2) and J (5), with a new skeleton have been isolated from the seeds of Garcinia subelliptica. Their structures, including relative configurations, were elucidated by spectroscopic methods and computer-generated molecular modeling. Compound 1 showed potent inhibitory effects on the release of beta-glucuronidase and lysozyme from rat neutrophils that had been stimulated with formyl-Met-Leu-Phe (fMLP)/cytochalasin B (CB). This effect was concentration-dependent with IC(50) values of 26.9+/-2.6 and 20.0+/-1.3 microM, respectively. Compound 1 also showed a potent concentration-dependent inhibitory effect on superoxide anion generation in rat neutrophils stimulated with fMLP/CB, with an IC(50) value of 17.0+/-0.9 microM. Compound 4 showed a potent inhibitory effect on NO production in culture media of N9 cells in response to lipopolysaccharide (LPS)/interferon-gamma (IFN-gamma) in a concentration-dependent manner with an IC(50) value of 7.4+/-0.2 microM.

Animals↗

Synthesis of new chiral bis(isoxazoline) ligands containing spiro[5.5]undecane skeleton.

New chiral bis(isoxazoline) ligands bearing a spiro[5.5]undecane skeleton were designed and synthesized in five steps from diethyl malonate (3). These ligands showed a coordinating ability to Cu(II) as chiral ligands. A complex of (+)-(M*,S*,R*)-[5.5]-SPRIX 2b and Cu(OTf)(2) catalyzed the conjugate addition of diethyl-zinc to 2-cyclohexenone (8) to give (S)-3-ethyl-cyclohexanone (9) in 93% yield with 54% ee.

Journal Article↗

Novel approach for the introduction of a C-1 oxygenated group on the decalin skeleton: first asymmetric total synthesis of 1S,6S-dihydroxy-eudesm-3-ene.

This paper describes a novel approach for the introduction of a C-1 hydroxyl on the decalin ring system, starting from (-)-carvone. Utilizing the substrate-controlled Mukaiyama aldol reaction and alkaline cyclization as key steps, the C-1 oxygenated decalin eudesmane skeleton 2' and its four isomers were synthesized efficiently. Furthermore, X-ray structural analysis showed that the claimed structure for the natural product is incorrect.

Biological Factors↗

The development and function of the skeleton and bone metastases.

Bone is a frequent site of metastases of the most common tumors, e.g., breast carcinoma and prostate carcinoma. The functions of the skeleton, calcium homeostasis and mechanical support, are carried out by the continuous destruction and rebuilding of small packets of this tissue called bone remodeling. Multinucleated, hemopoietically derived osteoclasts, which are related to macrophages, digest the bone, and mesenchymal-derived osteoblasts rebuild it. This process is kept in balance by finely regulated processes whereby osteoblast lineage cells respond to homeostatic signals and release factors that regulate osteoclast generation and activity. Cells that participate in inflammation and immunity also can stimulate osteoclast formation and lead to bone destruction. Tumor cells most likely subvert these physiologic processes to lodge in bone and cause metastases.

Bone Neoplasms↗

Expression patterns of Tgfbeta1-3 associate with myocardialisation of the outflow tract and the development of the epicardium and the fibrous heart skeleton.

Transforming growth factor-beta (Tgfbeta) is essential for normal embryogenesis. The cardiac phenotypes obtained after knockout of each of the three mammalian isoforms suggest different roles during morphogenesis. We studied cardiovascular expression of Tgfbeta1-3 in parallel tissue sections of normal mouse embryos from 9.5 to 15.5 days post coitum (dpc) by using radioactive in situ hybridisation. The Tgfbeta isoforms are differentially expressed in unique and in overlapping patterns during cardiovascular development. In the vessels, Tgfbeta1 is found in the intima, whereas Tgfbeta2 and -beta3 are mainly present in the media and adventitia of the great arteries. Tgfbeta1 is present in the endocardium at all stages examined. The Tgfbeta2 signal in the endocardium of the atrioventricular canal and outflow tract (9.5 dpc) shifts during epithelial-mesenchymal transformation toward the mesenchymal cushions (10.5-11.5 dpc) after which it exhibits a marked spatiotemporal expression pattern as the cushion differentiation progresses (11.5-15.5 dpc). The myocardium underlying the endocardial cushions and the atrial muscular septum are intensely positive for Tgfbeta2 at early stages (9.5-11.5 dpc) and expression decreases at 12.5 days. In contrast to earlier reports, we find marked overlap of Tgfbeta2 and -beta3 expression. Tgfbeta3 expression shows a characteristic distribution in the mesenchymal cushions, suggesting a role in cushion differentiation, possibly additional to Tgfbeta2. From 14.5 dpc onward, a strong Tgfbeta3 signal is found in the fibrous septum primum of the atrium and in the fibrous skeleton of the heart. Special attention was paid to the proepicardial organ and its derivatives. The proepicardial organ strongly expresses Tgfbeta2 as early as 9.5 days, and all isoforms are present in the epicardium from 12.5 dpc onward. The spatiotemporal cardiovascular expression of Tgfbeta1-3 supports both specific and complementary functions during cardiovascular development that might explain functional redundancy between the Tgfbeta-isoforms. The information provided favors novel roles of Tgfbeta1-3 in epicardial development, of Tgfbeta2 in myocardialisation, and of Tgfbeta3 in differentiation of the fibrous structures of the heart.

Actins↗

Kinetics of tamoxifen-regulated Cre activity in mice using a cartilage-specific CreER(T) to assay temporal activity windows along the proximodistal limb skeleton.

Cartilage differentiation occurs over a broad time range from early embryonic development, when the mesenchymal condensations that give rise to cartilage models for future bone first appear, and continuing through adult life, when there is ongoing maintenance of articular joint surfaces and re-activation of cartilage formation after fracture. The chondrogenic response also figures in the pathogenesis of degenerative and inflammatory joint diseases. We have generated a transgenic line expressing tamoxifen-dependent Cre recombinase that gives efficient recombination in the chondrogenic lineage, both during embryogenesis and postnatally, and provides a valuable tool for analysis of gene function selectively in chondrogenic cells using conditional genetic approaches. Because the cartilage model of the limb skeleton forms progressively in a proximodistal order during discrete, well-defined time periods, evaluation of the spatial extent of tamoxifen-induced recombination along the limb axis during these time windows has also enabled us to examine the pharmacokinetics of single-dose tamoxifen injections during pregnancy.

Animals↗

Mineral-bound noncollagenous proteins in archaeological human skeletons.

Archaeometric approaches to archaelogical human bone also include the extraction, identification and molecular analysis of surviving bone proteins. Due to its abundance as a matrix protein, most studies focus on collagen (e.g. radiocarbon dating). Also, a variety of serum proteins are detectable in excavated skeletons. Very limited knowledge still exists on mineral-bound noncollagenous bone proteins from ancient bones because, in the mature tissue, they occur in trace amounts only. Moreover, post-mortem decomposition is likely to change characteristic features of the molecules. Due to their suggested role as growth and developmental factors, identification and quantification of such proteins should be valuable for both physical anthropology and epidemiology. We present a valid method for the detection of small amounts of surviving mineral-bound noncollagenous proteins in excavated human bones up to 7500 years of age.

Archaeology↗

Effect of Bacillus Calmette-Guérin cell wall skeleton on the induction of the cytotoxic and suppressor T cells against syngeneic tumor in the mouse.

The effect of Bacillus Calmette-Guérin cell wall skeleton (BCG-CWS), which is one of the well-established immunopotentiators, on the induction of either cytotoxic or suppressor T cells against syngeneic tumor S1509a was investigated by an in vitro assay system. BCG-CWS showed an apparent augmenting activity in the induction of cytotoxic T cells against a syngeneic tumor when it was administered subcutaneously into mice with mitomycin C (MMC)-treated tumor cells in a form of oil-in-water emulsion. In addition, cytotoxic T cell activity was dominantly generated in regional lymph nodes. Furthermore, the immunized animals developed stronger resistance against the second inoculum of the same viable tumor cells. However, BCG-CWS had no effect on the induction of suppressor T cells in tumor-bearing host (TBH), and could not induce cytotoxic T cells against the homologous tumor in TBH in which suppressor T cells had been rapidly activated. The growth of tumor was neither suppressor nor enhanced, even if an oil-attached BCG-CWS was administered in TBH.

Animals↗

Regionalization of axial skeleton in the lungfish Neoceratodus forsteri (Dipnoi).

Differentiation of the axial skeleton into distinct regions, once thought to be characteristic of the Tetrapoda, also occurs in the actinopterygian Danio rerio. In these taxa, the boundary between the cervical-thoracic regions correlates with Hoxc6 expression and morphological features such as position of the pectoral fin and associated nerves, and the absence of ribs. In the lungfish Neoceratodus, a member of the extant sister taxon to the Tetrapoda, the first vertebral element to chondrify is situated well posterior to the skull, developing from somites 6 and 7 (6/7) and associated with an enlarged cranial rib and nerves innervating the pectoral fin. Two vertebral elements develop later and more anteriorly, associated with somites 4/5 and 5/6. These three elements become incorporated into the occipital region of the skull during Neoceratodus ontogeny, until the cranial rib itself articulates to the rear of the skull. These features of early development indicate a regionalization of the Neoceratodus vertebral column: the cranial rib marks the boundary between the cervical and thoracic regions, the two more anterior vertebrae lacking ribs represent the cervical region, while somites 1-4 (cranial half), lacking any vertebral development, represent the occipital region. However, the cervical region of the vertebral column is effectively lost during ontogeny of Neoceratodus. A recognizable cervical region in the tetrapod vertebral column, as in zebrafish, suggests that cervical vertebrae are not incorporated into the skull but maintained as distinct elements of the column, representing an important shift in relative developmental timing and the influence of heterochrony in this region during the fish-tetrapod transition.

Animals↗

Origination and innovation in the vertebrate limb skeleton: an epigenetic perspective.

The vertebrate limb has provided evolutionary and developmental biologists with grist for theory and experiment for at least a century. Its most salient features are its pattern of discrete skeletal elements, the general proximodistal increase in element number as development proceeds, and the individualization of size and shape of the elements in line with functional requirements. Despite increased knowledge of molecular changes during limb development, however, the mechanisms for origination and innovation of the vertebrate limb pattern are still uncertain. We suggest that the bauplan of the limb is based on an interplay of genetic and epigenetic processes; in particular, the self-organizing properties of precartilage mesenchymal tissue are proposed to provide the basis for its ability to generate regularly spaced nodules and rods of cartilage. We provide an experimentally based "core" set of cellular and molecular processes in limb mesenchyme that, under realistic conditions, exhibit the requisite self-organizing behavior for pattern origination. We describe simulations that show that under limb bud-like geometries the core mechanism gives rise to skeletons with authentic proximodistal spatiotemporal organization. Finally, we propose that evolution refines skeletal templates generated by this process by mobilizing accessory molecular and biomechanical regulatory processes to shape the developing limb and its individual elements. Morphological innovation may take place when such modulatory processes exceed a threshold defined by the dynamics of the skeletogenic system and elements are added or lost.

Animals↗

Postcranial pneumaticity: an evaluation of soft-tissue influences on the postcranial skeleton and the reconstruction of pulmonary anatomy in archosaurs.

Postcranial pneumaticity has been reported in numerous extinct sauropsid groups including pterosaurs, birds, saurischian dinosaurs, and, most recently, both crurotarsan and basal archosauriform taxa. By comparison with extant birds, pneumatic features in fossils have formed the basis for anatomical inferences concerning pulmonary structure and function, in addition to higher-level inferences related to growth, metabolic rate, and thermoregulation. In this study, gross dissection, vascular and pulmonary injection, and serial sectioning were employed to assess the manner in which different soft tissues impart their signature on the axial skeleton in a sample of birds, crocodylians, and lizards. Results from this study indicate that only cortical foramina or communicating fossae connected with large internal chambers are reliable and consistent indicators of pneumatic invasion of bone. As both vasculature and pneumatic diverticula may produce foramina of similar sizes and shapes, cortical features alone do not necessarily indicate pneumaticity. Noncommunicating (blind) vertebral fossae prove least useful, as these structures are associated with many different soft-tissue systems. This Pneumaticity Profile (PP) was used to evaluate the major clades of extinct archosauriform taxa with purported postcranial pneumaticity. Unambiguous indicators of pneumaticity are present only in certain ornithodiran archosaurs (e.g., sauropod and theropod dinosaurs, pterosaurs). In contrast, the basal archosauriform Erythrosuchus africanus and other nonornithodiran archosaurs (e.g., parasuchians) fail to satisfy morphological criteria of the PP, namely, that internal cavities are absent within bone, even though blind fossae and/or cortical foramina are present on vertebral neural arches. An examination of regional pneumaticity in extant avians reveals remarkably consistent patterns of diverticular invasion of bone, and thus provides increased resolution for inferring specific components of the pulmonary air sac system in their nonavian theropod ancestors. By comparison with well-preserved exemplars from within Neotheropoda (e.g., Abelisauridae, Allosauroidea), the following pattern emerges: pneumaticity of cervical vertebrae and ribs suggests pneumatization by lateral vertebral diverticula of a cervical air sac system, with sacral pneumaticity indicating the presence of caudally expanding air sacs and/or diverticula. The identification of postcranial pneumaticity in extinct taxa minimally forms the basis for inferring a heterogeneous pulmonary system with distinct exchange and nonexchange (i.e., air sacs) regions. Combined with inferences supporting a rigid, dorsally fixed lung, osteological indicators of cervical and abdominal air sacs highlight the fundamental layout of a flow-through pulmonary apparatus in nonavian theropods.

Abdomen↗

The fibrocartilage skeleton and related structures of the ventral pouch of balaenopterid whales.

The mandibular symphysis of rorqual whales, whales of the genera Megaptera and Balaenoptera, is characterized by a Y-shaped fibrocartilage structure that lies in the substance of the muscular ventral pouch of these animals. The stem of the structure joins with the symphysis and is usually indicated externally by an unfurrowed median strip of blubber that has been called the "cutwater" by earlier writers. The arms of the Y pass back and are superficially indicated in all rorqual whales as a ridge running parallel to the rami of the mandibles. This fibrocartilage skeleton of the pouch is most closely related to the mylohyoid muscle. The function of the fibrocartilage Y is probably linked with the jaw mechanics of these whales, but its precise function is otherwise not known.

Animals↗

Development of dentition and dermal skeleton in embryonic Scyliorhinus canicula.

Serial sections ranging from very young embryos to hatched juveniles and whole embryos of Scyliorhinus show that dentition and dermal skeleton belong to two independent secondary developmental fields that differ both developmentally and structurally. The development of the dentition starts very early, with a thickening of the ectoderm in the region of the mouth (stage 04), the invagination of the dental lamina (stage 18), and the formation of the germs of the first generation (stage 20). Tooth replacement movements start only near the end of embryogenesis (stage 35). Scale germs, on the other hand, first begin to form at stage 24. Scales erupt shortly before the animal hatches (stage 43). Only one scale generation is formed during embryogenesis. The forces which erupt the scales may come from fluid pressures in vacuoles of the fibrous layer of the dermis. Those which erupt the teeth probably also result from similar fluid pressures. The crown and upper part of the base of scales and teeth are formed by cells of the inner dental epithelium which are differentiated from the ectoderm. They are also formed by odontoblasts which are derived from the vascular layer of the dermis. However, the basal plates of scales and teeth containing the anchoring fibers are formed by osteoblasts, which are derived from the fibrous layer of the dermis.

Animals↗

Mass allometry of the appendicular skeleton in terrestrial mammals.

Most analyses on allometry of long bones in terrestrial mammals have focused on dimensional allometry, relating external bone measurements either to each other or to body mass. In this article, an analysis of long bone mass to body mass in 64 different species of mammals, spanning three orders of magnitude in body mass, is presented. As previously reported from analyses on total skeletal mass to body mass in terrestrial vertebrates, the masses of most appendicular bones scale with significant positive allometry. These include the pectoral and pelvic girdles, humerus, radius+ulna, and forelimb. Total hindlimb mass and the masses of individual hindlimb bones (femur, tibia, and metatarsus) scale isometrically. Metapodial mass correlates more poorly with body mass than the girdles or any of the long bones. Metapodial mass probably reflects locomotor behavior to a greater extent than do the long bones. Long bone mass in small mammals (<50 kg) scales with significantly greater positive allometry than bone mass in large (>50 kg) mammals, probably because of the proportionally shorter long bones of large mammals as a means of preserving resistance to bending forces at large body sizes. The positive allometric scaling of the skeleton in terrestrial animals has implications for the maximal size attainable, and it is possible that the largest sauropod dinosaurs approached this limit.

Animals↗

Primary tumors of the thoracic skeleton.

From 1975 to 1990, eighty-nine primary tumors of the thoracic skeleton; ribs, sternum, scapula, clavicle, and thoracic spine, were treated. Forty-four tumors (49%) were benign lesions. Forty-five tumors were malignant and were proportionately distributed amongst the five sites. The most common malignancies were Ewing's sarcomas, chondrosarcomas, plasmacytomas, osteogenic sarcomas, and lymphomas. All patients with Ewing's sarcomas were treated with combination chemotherapy, surgical resection, and radiation therapy for those with residual disease after surgery. Only one patient has died of disease. Patients with primary chondrosarcomas were treated by surgery alone and all are free of disease or have died without disease. Patients with solitary plasmacytomas or primary lymphomas of bone were treated with radiation therapy initially. Half the patients developed systemic disease. The patients with osteogenic sarcomas included several with radiation induced lesions and Paget's osteosarcoma and all but one died of disease.

Adult↗

Assessment of adult skeletons to detect prenatal exposure to 2,4,5-T or Trifluralin in mice.

A group of 88 spontaneously occurring variations of the skeleton were studies in adult CD-1 mice which had been exposed in utero to the herbicides 2,4,5-T or Trifluralin, administered to their dams by gavage, in an attempt to detect subtle biological effects of these compounds even in the absence of gross malformations. Sixty-seven characters showed variation in this population. Of these, 19 were significantly different in frequency from untreated controls in mice having receiving a teratogenic dose of 2,4,5-T, with an average difference of 23.7%. Seventeen of them were increased in frequency. By contrast, only three traits differed from untreated among mice receiving a no-effect dose of 2,4,5-T. Twelve traits differed significantly from untreated in the Trifluralin-treated group; the average difference was 18.8%, with 10 of the traits being increased in frequency. Two clusters of affected variants specific to the 2,4,5-T high dose group included frontal bone variants in the skull (presence of an interfrontal bone and fusion of the frontals) and variants in the cervical vertebrae (imperfect foramina in the first and second cervicals, dyssymphysis of the second cervical, and a shift of the arch foramen from the fourth to the fifth cervical). Two other effects peculiar to the 2,4,5-T-treated specimens were a loss of the prominent dorsal spine of the second thoracic vertebra and a reduction in the number of caudal vertebrae. The most obvious effects specific to the Trifluralin treatment were an increase in occurrence of 14 ribs, an undoubled foramen ovale, and the occurrence of accessory foramina in the cervical vertebrae. A striking increase in frequency of parted frontals was seen in both 2,4,5-T high dose and Trifluralin-treated groups. The skeletal variant assay system may be used as a postnatal screen for detecting prenatal exposure to potentially noxious substances.

2,4,5-Trichlorophenoxyacetic Acid↗

Distal phocomelia of the forearm in a thirteenth-century skeleton from Poland.

The skeleton of a 35- to 40-year-old male (A.D. 1250 to 1300) with distal phocomelia (intercalary transverse) of the right forearm and severe (90-100 degrees) right-sided scoliosis is examined. Congenital malformation of the the right forearm resulted in absence of the radius and ulna, and attachment of the upper arm and hand to the trunk. Lesser abnormalities consist of a-hypoplastic right scapula and misshapened ribs and sternum. This rare deformity reflects the variability and antiquity of congenital malformations of the upper limb.

Adult↗

Neural crest can form cartilages normally derived from mesoderm during development of the avian head skeleton.

The lateral wall of the avian braincase, which is indicative of the primitive amniote condition, is formed from mesoderm. In contrast, mammals have replaced this portion of their head skeleton with a nonhomologous bone of neural crest origin. Features that characterize the local developmental environment may have enabled a neural crest-derived skeletal element to be integrated into a mesodermal region of the braincase during the course of evolution. The lateral wall of the braincase lies along a boundary in the head that separates neural crest from mesoderm, and also, neural crest cells migrate through this region on their way to the first visceral arch. Differences in the availability of one skeletogenic population versus the other may determine the final composition of the lateral wall of the braincase. Using the quail-chick chimeric system, this investigation tests if populations of neural crest, when augmented and expanded within populations of mesoderm, will give rise to the lateral wall of the braincase. Results demonstrate that neural crest can produce cartilages that are morphologically indistinguishable from elements normally generated by mesoderm. These findings (1) indicate that neural crest can respond to the same cues that both promote skeletogenesis and enable proper patterning in mesoderm, (2) challenge hypotheses on the nature of the boundary between neural crest and mesoderm in the head, and (3) suggest that changes in the allocation of migrating cells could have enabled a neural crest-derived skeletal element to replace a mesodermal portion of the braincase during evolution.

Animals↗