Role of annexin 1 gene expression in mouse craniofacial bone development

BACKGROUND: Annexin 1 is a 37-kDa protein that has complex intra- and extracellular effects. To discover whether the absence of this protein alters bone development, we monitored this event in the annexin-A1 null mice in comparison with littermate wild-type controls. METHODS: Radiographic and densitometry methods were used for the assessment of bone in annexin-A1 null mice at a gross level. We used whole-skeleton staining, histological analysis, and Western blotting techniques to monitor changes at the tissue and cellular levels. RESULTS: There were no gross differences in the appendicular skeleton between the genotypes, but an anomalous development of the skull was observed in the annexin-A1 null mice. This was characterized in the newborn annexin-A1 null animals by a delayed intramembranous ossification of the skull, incomplete fusion of the interfrontal suture and palatine bone, and the presence of an abnormal suture structure. The annexin-A1 gene was shown to be active in osteocytes during this phase and COX-2 was abundantly expressed in cartilage and bone taken from annexin-A1 null mice. CONCLUSIONS: Expression of the annexin-A1 gene is important for the normal development of the skull in mice, possibly through the regulation of osteoblast differentiation and a secondary effect on the expression of components of the cPLA2-COX-2 system. © 2007 Wiley-Liss, Inc.

Premature suture closure and ectopic cranial bone in mice expressing Msx2 transgenes in the developing skull.

The coordinate growth of the brain and skull is achieved through a series of interactions between the developing brain, the growing bones of the skull, and the fibrous joints, or sutures, that unite the bones. These interactions couple the expansion of the brain to the growth of the bony plates at the sutures. Craniosynostosis, the premature fusion of the bones of the skull, is a common birth defect (1 in 3000 live births) that disrupts coordinate growth and often results in profoundly abnormal skull shape. Individuals affected with Boston-type craniosynostosis, an autosomal dominant disorder, bear a mutated copy of MSX2, a homeobox gene thought to function in tissue interactions. Here we show that expression of the mouse counterpart of this mutant gene in the developing skulls of transgenic mice causes craniosynostosis and ectopic cranial bone. These mice provide a transgenic model of craniosynostosis as well as a point of entry into the molecular mechanisms that coordinate the growth of the brain and skull.

Outer skull landmark-based coordinates for measurement of cerebral blood flow and intracranial pressure in rabbits

Despite the increased use of intracranial neuromonitoring during experimental subarachnoid hemorrhage (SAH), coordinates for probe placement in rabbits are lacking. This study evaluates the safety and reliability of using outer skull landmarks to identify locations for placement of cerebral blood flow (CBF) and intraparenchymal intracranial pressure (ICP) probes. Experimental SAH was performed in 17 rabbits using an extracranial-intracranial shunt model. ICP probes were placed in the frontal lobe and compared to measurements recorded from the olfactory bulb. CBF probes were placed in various locations in the frontal cortex anterior to the coronary suture. Insertion depth, relation to the ventricular system, and ideal placement location were determined by post-mortem examination. ICP recordings at the time of SAH from the frontal lobe did not differ significantly from those obtained from the right olfactory bulb. Ideal coordinates for intraparenchymal CBF probes in the left and right frontal lobe were found to be located 4.6±0.9 and 4.5±1.2 anterior to the bregma, 4.7±0.7mm and 4.7±0.5mm parasagittal, and at depths of 4±0.5mm and 3.9±0.5mm, respectively. The results demonstrate that the presented coordinates based on skull landmarks allow reliable placement of intraparenchymal ICP and CBF probes in rabbit brains without the use of a stereotactic frame.

A Wormian Bone, Mimicking an Entry Gunshot Wound of the Skull, in an Anthropological Specimen

Wormian bones (WB) are irregular small cranial ossicles found along suture lines and fontanels. In Brazil, gunshot wounds to the skull are quite common in young individuals. Nevertheless, as far as we know, this is the first report of a WB giving an erroneous aspect of gunshot entrance due to its displacement position. The present manuscript describes the case of a Brazilian young man who died due to ballistic trauma, where a gaping bony defect on the right side of the skull was thought to be the exit wound of an injury related to the destruction found on the left side, highly suggestive of firearm injury. Thus, this case study has brought to light similarities between a traumatic lesion and an orifice of a WB, with emphasis on differential diagnosis during routine anthropological examinations.

Glycosaminoglycan and growth factor mediated murine calvarial cell proliferation

Understanding the complex mechanisms underlying bone remodeling is crucial to the development of novel therapeutics. Glycosaminoglycans (GAGs) localised to the extracellular matrix (ECM) of bone are thought to play a key role in mediating aspects of bone development. The influence of isolated GAGs was studied by utilising in vitro murine calvarial monolayer and organ culture model systems. Addition of GAG preparations extracted from the cell surface of human osteoblasts at high concentrations (5 microg/ml) resulted in decreased proliferation of cells and decreased suture width and number of bone lining cells in calvarial sections. When we investigated potential interactions between the growth factors fibroblast growth factor-2 (FGF2), bone morphogenic protein-2 (BMP2) and transforming growth factor-beta1 (TGFbeta1) and the isolated cell surface GAGs, differences between the two model systems emerged. The cell culture system demonstrated a potentiating role for the isolated GAGs in the inhibition of FGF2 and TGFbeta1 actions. In contrast, the organ culture system demonstrated an enhanced stimulation of TFGbeta1 effects. These results emphasise the role of the ECM in mediating the interactions between GAGs and growth factors during bone development and suggest the GAG preparations contain potent inhibitory or stimulatory components able to mediate growth factor activity.

Galah's Skull

Poem

Retrogressed eclogite (20 kbar, 1020 °C) from the Neoproterozoic Palghat–Cauvery suture zone, southern India

The Palghat–Cauvery suture zone in southern India separates Archaean crustal blocks to the north and the Proterozoic Madurai block to the south. Here we present the first detailed study of a partially retrogressed eclogite (from within the Sittampundi anorthositic complex in the suture zone) that occurs as a 20-cm wide layer in a garnet gabbro layer in anorthosite. The eclogite largely consists of an assemblage of coexisting porphyroblasts of almandine–pyrope garnet and augitic clinopyroxene. However, a few garnets contain inclusions of omphacite. Rims and symplectites composed of Na–Ca amphibole and plagioclase form a retrograde assemblage. Petrographic analysis and calculated phase equilibria indicate that garnet–omphacite–rutile–melt was the peak metamorphic assemblage and that it formed at ca. 20 kbar and above 1000 °C. The eclogite was exhumed on a very tight hairpin-type, anticlockwise P–T path, which we relate to subduction and exhumation in the Palghat–Cauvery suture zone. The REE composition of the minerals suggests a basaltic oceanic crustal protolith metamorphosed in a subduction regime. Geological–structural relations combined with geophysical data from the Palghat–Cauvery suture zone suggest that the eclogite facies metamorphism was related to formation of the suture zone. Closure of the Mozambique Ocean led to development of the suture zone and to its western extension in the Betsimisaraka suture of Madagascar.

An evaluation of genetic analyses, skull morphology and visual appearance for assessing dingo purity: implications for dingo conservation

The introgression of domestic dog genes into dingo populations threatens the genetic integrity of 'pure' dingoes. However, dingo conservation efforts are hampered by difficulties in distinguishing between dingoes and hybrids in the field. This study evaluates consistency in the status of hybridisation (i.e. dingo, hybrid or dog) assigned by genetic analyses, skull morphology and visual assessments. Of the 56 south-east Queensland animals sampled, 39 (69.6%) were assigned the same status by all three methods, 10 (17.9%) by genetic and skull methods, four (7.1%) by genetic and visual methods; and two (3.6%) by skull and visual methods. Pair-wise comparisons identified a significant relationship between genetic and skull methods, but not between either of these and visual methods. Results from surveying 13 experienced wild dog managers showed that hybrids were more easily identified by visual characters than were dingoes. A more reliable visual assessment can be developed through determining the relationship between (1) genetics and phenotype by sampling wild dog populations and (2) the expression of visual characteristics from different proportions and breeds of domestic dog genes by breeding trials. Culling obvious hybrids based on visual characteristics, such as sable and patchy coat colours, should slow the process of hybridisation.

K-Ar ages of biotite and muscovite from Pangong metamorphic complex, Shyok suture zone, India: Implications for the youngest post-collision metamorphic event in Ladakh Himalaya

The garnet-kyanite-staurolite and garnet-biotite-staurolite gneisses were collected from a locality within Lukung area that belongs to the Pangong metamorphic complex in Shyok valley, Ladakh Himalaya. The kyanite-free samples have garnet and staurolite in equilibrium, where garnets show euhedral texture and have flat compositional profile. On the other hand, the kyanite-bearing sample shows equilibrium assemblage of garnet-kyanite-staurolite along with muscovite and biotite. In this case, garnet has an inclusion rich core with a distinct grain boundary, which was later overgrown by inclusion free euhedral garnet. Garnet cores are rich in Mn and Ca, while the rims are poor in Mn and rich in Fe and Mg, suggesting two distinct generations of growth. However, the compositional profiles and textural signature of garnets suggests the same stage of P -T evolution for the formation of the inclusion free euhedral garnets in the kyanite-free gneisses and the inclusion free euhedral garnet rims in the kyanite-bearing gneiss. Muscovites from the four samples have consistent K-Ar ages, suggesting the cooling age (∼ 10 Ma) of the gneisses. These ages make a constraint on the timing of the youngest post-collision metamorphic event that may be closely related to an activation of the Karakoram fault in Pangong metamorphic complex.

Development of a new flexor tendon repair technique performed with bioabsorbable poly-L/D-lactide (PLDLA) suture

Sormen koukistajajännevamman korjauksen jälkeisen aktiivisen mobilisaation on todettu johtavan parempaan toiminnalliseen lopputulokseen kuin nykyisin yleisesti käytetyn dynaamisen mobilisaation. Aktiivisen mobilisaation ongelma on jännekorjauksen pettämisriskin lisääntyminen nykyisten ommeltekniikoiden riittämättömän vahvuuden vuoksi. Jännekorjauksen lujuutta on parannettu kehittämällä monisäieommeltekniikoita, joissa jänteeseen tehdään useita rinnakkaisia ydinompeleita. Niiden kliinistä käyttöä rajoittaa kuitenkin monimutkainen ja aikaa vievä tekninen suoritus. Käden koukistajajännekorjauksessa käytetään yleisesti sulamattomia ommelmateriaaleja. Nykyiset käytössä olevat biohajoavat langat heikkenevät liian nopeasti jänteen paranemiseen nähden. Biohajoavan laktidistereokopolymeeri (PLDLA) 96/4 – langan vetolujuuden puoliintumisajan sekä kudosominaisuuksien on aiemmin todettu soveltuvan koukistajajännekorjaukseen. Tutkimuksen tavoitteena oli kehittää välittömän aktiivisen mobilisaation kestävä ja toteutukseltaan yksinkertainen käden koukistajajännekorjausmenetelmä biohajoavaa PLDLA 96/4 –materiaalia käyttäen. Tutkimuksessa analysoitiin viiden eri yleisesti käytetyn koukistajajänneompeleen biomekaanisia ominaisuuksia staattisessa vetolujuustestauksessa ydinompeleen rakenteellisten ominaisuuksien – 1) säikeiden (lankojen) lukumäärän, 2) langan paksuuden ja 3) ompeleen konfiguraation – vaikutuksen selvittämiseksi jännekorjauksen pettämiseen ja vahvuuteen. Jännekorjausten näkyvän avautumisen todettiin alkavan perifeerisen ompeleen pettäessä voima-venymäkäyrän myötöpisteessä. Ydinompeleen lankojen lukumäärän lisääminen paransi ompeleen pitokykyä jänteessä ja suurensi korjauksen myötövoimaa. Sen sijaan paksumman (vahvemman) langan käyttäminen tai ompeleen konfiguraatio eivät vaikuttaneet myötövoimaan. Tulosten perusteella tutkittiin mahdollisuuksia lisätä ompeleen pitokykyä jänteestä yksinkertaisella monisäieompeleella, jossa ydinommel tehtiin kolmen säikeen polyesterilangalla tai nauhamaisen rakenteen omaavalla kolmen säikeen polyesterilangalla. Nauhamainen rakenne lisäsi merkitsevästi ompeleen pitokykyä jänteessä parantaen myötövoimaa sekä maksimivoimaa. Korjauksen vahvuus ylitti aktiivisen mobilisaation jännekorjaukseen kohdistaman kuormitustason. PLDLA 96/4 –langan soveltuvuutta koukistajajännekorjaukseen selvitettiin tutkimalla langan biomekaanisia ominaisuuksia ja solmujen pito-ominaisuuksia staattisessa vetolujuustestauksessa verrattuna yleisimmin jännekorjauksessa käytettävään punottuun polyesterilankaan (Ticron®). PLDLA –langan todettiin soveltuvan hyvin koukistajajännekorjaukseen, sillä se on polyesterilankaa venymättömämpi ja solmujen pitävyys on parempi. Viimeisessä vaiheessa tutkittiin PLDLA 96/4 –langasta valmistetulla kolmisäikeisellä, nauhamaisella jännekorjausvälineellä tehdyn jännekorjauksen kestävyyttä staattisessa vetolujuustestauksessa sekä syklisessä kuormituksessa, joka simuloi staattista testausta paremmin mobilisaation toistuvaa kuormitusta. PLDLA-korjauksen vahvuus ylitti sekä staattisessa että syklisessä kuormituksessa aktiivisen mobilisaation edellyttämän vahvuuden. Nauhamaista litteää ommelmateriaalia ei aiemmin ole tutkittu tai käytetty käden koukistajajännekorjauksessa. Tässä tutkimuksessa ommelmateriaalin nauhamainen rakenne paransi merkitsevästi jännekorjauksen vahvuutta, minkä arvioidaan johtuvan lisääntyneestä kontaktipinnasta jänteen ja ommelmateriaalin välillä estäen ompeleen läpileikkautumista jänteessä. Tutkimuksessa biohajoavasta PLDLA –materiaalista valmistetulla rakenteeltaan nauhamaisella kolmisäikeisellä langalla tehdyn jännekorjauksen vahvuus saavutti aktiivisen mobilisaation edellyttämän tason. Lisäksi uusi menetelmä on helppokäyttöinen ja sillä vältetään perinteisten monisäieompeleiden tekniseen suoritukseen liittyvät ongelmat.

A Rodinian suture in western India: New insights on India-Madagascar correlations

We report detailed evidence for a new paleo-suture zone (the Kumta suture) on the western margin of southern India. The c. 15-km-wide, westward dipping suture zone contains garnet-biotite, fuchsite-haematite, chlorite-quartz, quartz-phengite schists, biotite augen gneiss, marble and amphibolite. The isochemical phase diagram estimations and the high-Si phengite composition of quartz-phengite schist suggest a near-peak condition of c. 18 kbar at c. 550 degrees C, followed by near-isothermal decompression. The detrital SHRIMP U-Pb zircon ages from quartz-phengite schist give four age populations ranging from 3280 to 2993 Ma. Phengite from quartz-phengite schist and biotite from garnet-biotite schist have K-Ar metamorphic ages of ca. 1326 and ca. 1385 Ma respectively. Electron microprobe-CHIME ages of in situ zircons in quartz-phengite schist (ca. 3750 Ma and ca. 1697 Ma) are consistent with the above results. The Bondla ultramafic-gabbro complex in the west of the Kumta suture compositionally represents an arc with K-Ar biotite ages from gabbro in the range 1644-1536 Ma. On the eastern side of the suture are weakly deformed and unmetamorphosed shallow westward-dipping sedimentary rocks of the Sirsi shelf, which has the following upward stratigraphy: pebbly quartzite/sandstone, turbidite, magnetite iron formation, and limestone; farther east the lower lying quartzite has an unconformable contact with ca. 2571 Ma quartzo-feldspathic gneisses of the Dharwar block with a ca. 1733 Ma biotite cooling age. To the west of the suture is a c. 60-km-wide Karwar block mainly consisting of tonalite-trondhjemite-granodiorite (TTG) and amphibolite. The TTGs have U-Pb zircon magmatic ages of ca. 3200 Ma with a rare inherited core age of ca. 3601 Ma. The K-Ar biotite cooling age from the TTGs (1746 Ma and 1796 Ma) and amphibolite (ca. 1697 Ma) represents late-stage uplift. Integration of geological, structural and geochronological data from western India and eastern Madagascar suggest diachronous ocean closure during the amalgamation of Rodinia; in the north at around ca. 1380 Ma, and a progression toward the south until ca. 750 Ma. Satellite imagery based regional structural lineaments suggests that the Betsimisaraka suture continues into western India as the Kumta suture and possibly farther south toward a suture in the Coorg area, representing in total a c. 1000 km long Rodinian suture. (C) 2013 Elsevier B.V. All rights reserved.