Vertebral Sites Research Articles

Ten percent of conserved miRNA-binding sites in vertebrates are misaligned

MiRNAs are small endogenous noncoding RNAs that inhibit protein expression at the translation level via complementary binding to a specific site in the mRNA. Most of the functional binding sites (miRNA nucleotides 2–7) located within the 3' untranslated region are conserved. These sites are predicted using the methods of comparative genomics, primarily, multiple sequence alignment. However, multiple sequence alignments are prone to accumulating errors due to strong divergence of species. Moreover, in the course of evolution, binding sites can migrate along the sequence. The aim of this work was to estimate the portion of conserved miRNA-binding sites that cannot be predicted using the existing tools because of these phenomena. The concept of L-conserved sites is introduced: a site is termed L-conserved if it is present in each sequence in the alignment within a frame of length L. We observed a significant increase in the number of additionally detected conserved sites without loss of sensitivity. The effect of species divergence on this increase was also evaluated.

Research of the effect of bone mineral density and fracture site of the vertebrae on low back pain in elderly patients with osteoporotic vertebral compression fractures

To investigate the effect of bone mineral density(BMD) and fracture site of the vertebrae on low back pain in elderly patients with osteoporotic vertebral compression fractures. From August 2011 to August 2013, a total of 107 senile patients with osteoporotic vertebral compression fractures underwent percutaneous vertebroplasty were followed up for more than 2 ( average 2.5) years in Department of orthopedics, Xuanwu Hospital, Capital Medical University. The incidence of low back pain after vertebroplasty were analyzed on visual analog scale (VAS), and the relationship between BMD, vertebral fracture site were investigated. A total of 18 cases(16.8%)after vertebroplasty have significant low back pain. Along with BMD decreased [(-2.90±0.91) vs (-4.87±0.52)], the VAS of low back pain increasing, which showed statistical significance difference[(-3.12±0.91) vs (4.03±1.08), P<0.05]. The site of vertebral fracture was lower, the VAS was higher. About 16.8% patients show obvious low back pain after vertebroplasty. BMD and Vertebral fracture site were important factors of low back pain in elderly patients with osteoporotic vertebral compression fractures after vertebroplasty.

THE ANATOMY OF PEDIATRIC LAPAROSCOPIC AND THORACOSCOPIC PROCEDURES ‐ NEWER CONCEPTS AND PROCEDURES

Laparoscopic procedures in pediatric surgery can be be divided grossly into abdominal and thoracic procedures. The basic procedures in laparoscopy require an understanding of body wall surface and muscle anatomy. Laparoscopic pediatric procedures have revolutionized the treatment of tracheo‐oesophageal fistulas with oesophageal atresia, duodenal atresia, anorectal malformations, appendicitis, hirschsrpung disease and inguinal hernias.Abdominal laparoscopy requires gas insufflation, the use of surface ports in various sites and and a keen understanding of upper and lower abdominal anatomy. Iatrogenic abdominal injuries include vascular trauma, retroperitoneal heamorrhage, bladder injuries and stomach injuries especially when it is overinflated. Most procedures use a camera port with 2 or more ports for instrumentation. Lower abdominal laparoscopies have a risk of hernias, damage to the inferior epigastric blood vessels and bowel damage. A detailed knowledge of the abdominal body wall musculature and underlying vascular structures is required. The anatomy reinforcing single port laparoscopy procedures is presented in this work too. Transuterine and cul‐de‐sac approaches are revisited and explained.Thoracospic pediatric surgical procedures have a risk of injury to the lung, pleura and thoracic wall vasculature. Newer anatomical approaches through the posterior thoracic wall ports are also proposed and explained.To correct spinal and vertebral defects newer sites of laparoscopic port entry are proposed with an eye to access and maximum utilization of small working space in such procedures.Support or Funding InformationFunding received from the Ministry of Health, Saudi Arabia and the Indian Institute of Morphology, Kolkata.

Expanding the diversity of DNA base modifications with N⁶-methyldeoxyadenosine.

Vertebrate DNA is subjected to epigenetic base modifications that have been thought to be limited to methylated and other modified forms of cytidine. A recent study shows that methylation of adenine to form N6-methyladenine is a rare but readily detectable modification that can be mapped to distinct genomic sites in vertebrates.

Syringe-pressurised bone wax to stop bleeding from vertebral body pin sites during anterior cervical discectomy and fusion (ACDF).

Anterior cervical discectomy and fusion (ACDF) is a common spinal procedure, with over 500,000 cases performed between 1990 and 1999 in the USA alone. ACDF is well-established for the management of patients with radiculopathy and/or myelopathy secondary to degenerative disc disease and/or spondylosis. In 2007, Fountas et al reported a postoperative haematoma rate of 5.6% (n=57) in a series of 1015 ACDFs, with emergent surgical evacuation of a haematoma required in 2.4% (n=24) of cases. No patients who developed haematomas had a bleeding diathesis and, at the time of evacuation, no active bleeding points were seen. Many surgeons place distraction pins in the vertebrae above and below the disc to facilitate the procedure (Figure 1). In our experience, removal of these pins towards the end of the procedure can lead to bleeding from the resultant holes that can be hard to control (Figure 2). We present a technique to stop bleeding from the vertebral body pin sites via the use of syringe-pressurised bone wax.

Underdiagnosis of osteoporotic vertebral fractures in patients with fragility fractures: retrospective analysis of over 300 patients.

Osteoporosis (OP) is a silent disease unless a fracture occurs; it is a major health problem, mainly due to fragility fractures, that occur at vertebral and peripheral sites. Vertebral fractures (VF) are probably the most common fragility fractures, but they go often unrecognized. The main clinical symptoms of VF are acute and chronic back pain, spinal deformity, reduced mobility and impaired quality of life. They are frequently associated with other fragility fractures. We examined 478 patients at our outpatient clinic, who were referred for fragility fracture occurrence. The most common fragility fractures was hip fractures. However, after execution of spine X-rays in patients who had sustained hip fracture, we found that a large proportion of them had VF, which had not been reported in their medical history.

Identification and Expansion of Adipose Stem Cells with Enhanced Bone Regeneration Properties

Introduction: Adipose tissue represents an abundant source of mesenchymal stem-like cells. Adipose-derived stem cell progeny have been investigated and used in regenerative medicine for decades. In the last few years, they have been used to “enrich” lipoaspirates in fat grafting techniques, in an attempt to boost the regenerative potential of adipose tissue when used as autologous “filler”. Materials and Methods: 50 adipose tissue samples from lipoaspirates and subcutaneous breast tissue biopsies were used to generate adipose floating spheroid cell (ASphC) lines. ASphC were characterized for the expression of putative mesenchymal stem cell markers and used in vitro to test their multilineage potential. Furthermore, ASphCs were seeded on dermal regeneration template (Integra®) and implanted into the T8 vertebral laminectomy site of immunocompromised mice. Results: Here, we show that the majority of ASphCs are in a quiescent state and express the putative surface stem cell marker CD271. Unlike CD271- cells, CD271+ ASphCs grew indefinitely in vitro as undifferentiated spheres in serum-free medium, maintaining their multilineage differentiation potential ability. Importantly, p107, a functional adipose stem cell marker, is strictly expressed in ASphCs and barely present in their differentiated mesenchymal lineages. These sphere cells display an enhanced ability in vitro to differentiate into distinctive end-stage cell types, such as osteoblasts, chondrocytes and adipocytes. Gene expression profiling analysis indicated that ASphCs are endowed with stem cell potential that is gradually lost during specific differentiation. Finally, ASphCs facilitate bone inter body repair and regeneration after laminectomy. Conclusions: We conclude that ASphCs possess a pronounced in vivo activity to regenerate the bone injury. Overall, ASphCs represent a heterogeneous population of stem-like cells harbouring multilineage differential potential and representing a prospective promising tool in cell therapy and tissue engineering.

A review of the upper Campanian vertebrate site of Armuña (Segovia Province, Spain)

The Upper Cretaceous outcrops of Armuña (Segovia Province, Spain) yielded relatively abundant material of vertebrates during prospection and excavation in the second half of the 1980s. However, little has been published on these remains. A new analysis of the specimens from this upper Campanian site reveals the presence of some clades in the site for the first time (e.g., Dortokidae, Anguimorpha, Mosasauroidea). Furthermore, the material of the clades previously recognized there has been reviewed and described in more detail, with some previous systematic attributions confirmed and others refuted. Consequently, a relatively high local diversity has been identified. New taxa (i.e., a member of Anguimorpha and a eusuchian crocodyliform) are identified in Armuña, coexisting with other taxa previously described in other sites from the Iberoarmorican Realm. The vertebrates from Armuña confirm that the fauna from the Upper Cretaceous of the Iberian Peninsula is composed of a mixture of European endemic clades and lineages shared with other continents such as North America (e.g., anguimorphs) and Africa (e.g., bothremydids).

Magnetostratigraphy and lithostratigraphy of the Laño vertebrate-site: Implications in the uppermost Cretaceous chronostratigraphy of the Basque-Cantabrian Region

Here we present the magnetostratigraphic dating of the Laño locality (Condado de Treviño, northern Iberian Peninsula), one of the most noteworthy Campanian-Maastrichtian vertebrate sites of Europe. A composite section of 75 m thickness (Laño quarry) constructed from multiple, overlapping profiles and a continuous one (Faido) have been sampled for magnetostratigraphy. Thermal demagnetization techniques were systematically applied to 161 standard specimens and allowed characterizing the characteristic remanent magnetism, mostly carried out by magnetite. The palaeomagnetic signal is slightly scattered due to variety of lithologies, but the primary character can be guaranteed, since the normal and reverse directions are pseudo antiparallel; 346, 28 (α95: 11.9°, k: 5.3) and 175, −35 (α95: 16.4°, k: 4.6). Reliable samples allowed us to build the local polarity sequence made of eight magnetozones that has been used to correlate to the Global Polarity Time Scale. The age of the lower part of the Laño-village succession is basal late Campanian (Hoplitoplacenticeras marroti ammonite zone) and fits with the long reversed zone that must correlate to Chron C33r. The pattern of magnetozones allows tracking the section up to C30r at the upper part of the profile. In this correlation, the Laño vertebrate site is regarded as latest Campanian in age as it falls within the C32n (≈72–73.5 Ma). The combined lithostratigraphic and magnetostratigraphic analyses have yielded additional conclusions regarding the vertebrate assemblages that are representative of the Late Campanian of the Iberian Peninsula, in addition to highlight an older occurrence in Europe of some vertebrate groups such as salamandrid lissamphians and anguid lizards (or amphisbaenians).

Elevated incidence of fractures in women with invasive breast cancer.

This study evaluates the incidence of bone fractures in women with BC.We found that women with invasive breast cancer are at an increased risk for bone fractures, with fractures most commonly occurring at lower extremity and vertebral sites. The risk is further increased in women undergoing cancer therapy. Bone loss and fractures in breast cancer have generally been attributed to aromatase inhibitor use. This study assessed the incidence of fractures after invasive breast cancer diagnosis and evaluated bone density and FRAX risk calculation at time of fracture occurrence. Retrospective cohort study of women with invasive breast cancer [June 2003-December 2011] who participated in an academic hospital based genetic biobank. Demographic and clinical characteristics were abstracted from the electronic medical record (EMR). A total of 422 women with invasive breast cancer were assessed; 79 (28%) sustained fractures during the observation period; fractures occurred at multiple skeletal sites in 27 cases (116 fractures). The incidence of fractures was 40 per 1000 person-years. Women who sustained fractures were mostly white and had a family history of osteoporosis (36.9%, p = 0.03) or history of a prior fracture (6/79, p = 0.004). Fractures occurred 4.0years (range 0-12years) after cancer diagnosis. Fracture cases had femoral neck bone mineral density (BMD) of 0.72 + 0.12g/cm(2), T-score of -1.2, that is, within the low bone mass range. Fractures most commonly occurred in lower extremities, vertebral, and wrist sites. Hip fractures accounted for 11% of fractures, occurring at a median age of 61years. Fractures occur shortly after commencing cancer therapy. Rapid bone loss associated with cancer therapy may precipitate fractures. Fractures occur at relatively higher BMD in BC. Occurrence of fractures in invasive breast cancer raises the possibility of cancer-induced impairment in bone quality.

Mutations in Nature Conferred a High Affinity Phosphatidylinositol 4,5-Bisphosphate-binding Site in Vertebrate Inwardly Rectifying Potassium Channels

All vertebrate inwardly rectifying potassium (Kir) channels are activated by phosphatidylinositol 4,5-bisphosphate (PIP2) (Logothetis, D. E., Petrou, V. I., Zhang, M., Mahajan, R., Meng, X. Y., Adney, S. K., Cui, M., and Baki, L. (2015) Annu. Rev. Physiol. 77, 81-104; Fürst, O., Mondou, B., and D'Avanzo, N. (2014) Front. Physiol. 4, 404-404). Structural components of a PIP2-binding site are conserved in vertebrate Kir channels but not in distantly related animals such as sponges and sea anemones. To expand our understanding of the structure-function relationships of PIP2 regulation of Kir channels, we studied AqKir, which was cloned from the marine sponge Amphimedon queenslandica, an animal that represents the phylogenetically oldest metazoans. A requirement for PIP2 in the maintenance of AqKir activity was examined in intact oocytes by activation of a co-expressed voltage-sensing phosphatase, application of wortmannin (at micromolar concentrations), and activation of a co-expressed muscarinic acetylcholine receptor. All three mechanisms to reduce the availability of PIP2 resulted in inhibition of AqKir current. However, time-dependent rundown of AqKir currents in inside-out patches could not be re-activated by direct application to the inside membrane surface of water-soluble dioctanoyl PIP2, and the current was incompletely re-activated by the more hydrophobic arachidonyl stearyl PIP2. When we introduced mutations to AqKir to restore two positive charges within the vertebrate PIP2-binding site, both forms of PIP2 strongly re-activated the mutant sponge channels in inside-out patches. Molecular dynamics simulations validate the additional hydrogen bonding potential of the sponge channel mutants. Thus, nature's mutations conferred a high affinity activation of vertebrate Kir channels by PIP2, and this is a more recent evolutionary development than the structures that explain ion channel selectivity and inward rectification.

Draper/CED-1 Mediates an Ancient Damage Response to Control Inflammatory Blood Cell Migration In Vivo

SummaryTissue damage leads to a robust and rapid inflammatory response whereby leukocytes are actively drawn toward the wound. Hydrogen peroxide (H2O2) has been shown to be an immediate damage signal essential for the recruitment of these inflammatory blood cells to wound sites in both Drosophila and vertebrates [1, 2]. Recent studies in zebrafish have shown that wound-induced H2O2 is detected by the redox-sensitive Src family kinase (SFK) Lyn within the responding blood cells [3]. Here, we show the same signaling occurs in Drosophila inflammatory cells in response to wound-induced H2O2 with mutants for the Lyn homolog Src42A displaying impaired inflammatory migration to wounds. We go on to show that activation of Src42A is necessary to trigger a signaling cascade within the inflammatory cells involving the ITAM domain-containing protein Draper-I (a member of the CED-1 family of apoptotic cell clearance receptors) and a downstream kinase, Shark, that is required for migration to wounds. The Src42A-Draper-Shark-mediated signaling axis is homologous to the well-established SFK-ITAM-Syk-signaling pathway used in vertebrate adaptive immune responses. Consequently, our results suggest that adaptive immunoreceptor-signaling pathways important in distinguishing self from non-self appear to have evolved from a more-ancient damage response. Furthermore, this changes the role of H2O2 from an inflammatory chemoattractant to an activator signal that primes immune cells to respond to damage cues via the activation of damage receptors such as Draper.

Teleost soluble CSF-1R modulates cytokine profiles at an inflammatory site, and inhibits neutrophil chemotaxis, phagocytosis, and bacterial killing

Soluble colony stimulating factor-1 receptor (sCSF-1R) is a novel bony fish protein that contributes to the regulation of macrophage proliferation. We recently showed that this soluble receptor is highly upregulated by teleost macrophages in the presence of apoptotic cells. Further, recombinant sCSF-1R inhibited leukocyte infiltration into a challenge site in vivo. Herein, we characterized the mechanisms underlying these changes as a platform to better understand the evolutionary origins of the CSF-1 immune-regulatory axis and inflammation control in teleosts. Using an in vivo model of self-resolving peritonitis, we show that sCSF-1R downregulates chemokine expression and inhibits neutrophil chemotaxis. Soluble CSF-1R also inhibited gene expression of several pro-inflammatory cytokines and promoted the expression of an anti-inflammatory mediator, IL-10. Finally, the phenotype of infiltrating neutrophils changed significantly in the presence of sCSF-1R. Both a reduced capacity for phagocytosis and pathogen killing were observed. Overall, our results implicate sCSF-1R as an important regulator of neutrophil responses in teleosts. It remains unclear whether this represents an inflammation regulatory factor that is unique to this animal group or one that may be evolutionarily conserved and continues to contribute to the regulation of antimicrobial processes at inflammatory sites in higher vertebrates.

Rivers as repositories for fossil vertebrates: a case study from the Upper Cretaceous of southern France

An accumulation of vertebrate remains in Late Cretaceous continental deposits at Cruzy (Hérault, south-western France) is studied, using sedimentological (clay mineralogy, granulometry, cathodoluminescence) and taphonomic approaches, in order to reconstruct the depositional environment under which it was formed. The fossil-bearing conglomerates, sands and clays were apparently deposited during brief flood episodes affecting a braided river system, under a “tropical” type of climate with alternating dry and wet seasons. This type of depositional environment shows close similarities with those reconstructed for Late Cretaceous vertebrate sites in Romania.

Fossil assemblages and palaeoenvironments in the Cenomanian vertebrate site of Nazaré (West Central Portugal)

Fossil assemblages and palaeoenvironments in the Cenomanian vertebrate site of Nazaré (West Central Portugal)

Bone Accumulation by Leopards in the Late Pleistocene in the Moncayo Massif (Zaragoza, NE Spain)

Eating habits of Panthera pardus are well known. When there are caves in its territory, prey accumulates inside them. This helps to prevent its kill from being stolen by other predators like hyenas. Although the leopard is an accumulator of bones in caves, few studies have been conducted on existing lairs. There are, however, examples of fossil vertebrate sites whose main collecting agent is the leopard. During the Late Pleistocene, the leopard was a common carnivore in European faunal associations. Here we present a new locality of Quaternary mammals with a scarce human presence, the cave of Los Rincones (province of Zaragoza, Spain); we show the leopard to be the main accumulator of the bones in the cave, while there are no interactions between humans and leopards. For this purpose, a taphonomic analysis is performed on different bone-layers of the cave.

COMPARISON BETWEEN COMPUTED TOMOGRAPHY, FLUOROSCOPY, AND ULTRASONOGRAPHY FOR GUIDING PERCUTANEOUS INJECTION OF THE CANINE INTERVERTEBRAL DISC

A minimally invasive method for delivering injectable therapeutic agents would be desirable for the treatment of intervertebral disc disease in dogs. The purpose of this study was to compare computed tomography (CT), ultrasonography (US), and fluoroscopy modalities for guiding percutaneous injection into canine intervertebral discs. Intervertebral discs of 14 dog cadavers were injected with a gelified ethanol therapeutic agent. Successful injectate placement and injectate leakage were determined based on necropsy inspection of discs. Injection into the nucleus pulposus was successful in 55 of 78 (71%) of all injected discs. Injections guided using CT and fluoroscopy were significantly more successful than US-guided injections. Odds of successful injection without leakage were greater for CT vs. US (P = 0.0026) but there was no significant difference between CT and fluoroscopy (P = 0.0620). Injection success rates did not differ among vertebral sites or dog cadavers of varying weights. Forty-nine (63%) of injection sites had injectate leakage outside the disc and 10 of these involved structures within the vertebral canal. The highest rate of injection success with the least amount of leakage was achieved with CT guidance. Findings indicated that CT, fluoroscopy, and US are feasible modalities for guiding percutaneous injection of a gelified ethanol therapeutic agent into the canine intervertebral disc, with moderate to high success rates for different regions of the spine. However, a moderately high rate of injectate leakage occurred outside of the disc and this should be taken into consideration for future safety and efficacy studies.

Petreşti-Arini – An important but ephemeral Upper Cretaceous continental vertebrate site in the southwestern Transylvanian Basin, Romania

The Transylvanian region of Romania preserves some of the most unusual and iconic dinosaurs in the global fossil record, including dwarfed herbivores and aberrant carnivores that lived during the very latest Cretaceous (Maastrichtian) in an ancient island ecosystem (the Haţeg Island). A series of artificial outcrops recently exposed during a hydroelectric project, the Petreşti-Arini section near Sebeş in the Transylvanian Basin, records a 400+ meter sequence documenting the transition from fully marine to terrestrial environments during the Campanian–Maastrichtian. Calcareous nannofossil biostratigraphy indicates that the lower marine beds in this section, part of the uppermost Bozeş Formation, can be assigned to the CC22 biozone, corresponding to the lower–mid upper Campanian. These beds smoothly transition, via a brackish-water unit, into the fully continental Maastrichtian Sebeş Formation. Dinosaur and pterosaur fossils from the uppermost Bozeş Formation can be assigned a late Campanian age making them the oldest well-dated terrestrial fossils from the Haţeg Island, and indicating that the classic Haţeg dinosaur fauna was becoming established by this time, coincident with the first emergence of widespread land areas. Vertebrate fossils occur throughout the overlying Sebeş Formation at the site and are dominated by the small-bodied herbivorous dinosaur Zalmoxes. The dominance of Zalmoxes, and the absence of many taxa commonly seen elsewhere in Maastrichtian sites in Romania, suggests the possibility that either the Petreşti-Arini section preserves a somewhat unusual near-shore environment, or the earliest Haţeg Island dinosaur communities were structured differently from the more diverse communities later in the Maastrichtian. Alternatively, due to the limited sample size available from the studied succession, it is also conceivable that sampling biases give an incomplete portrayal of the Petreşti-Arini local fauna. Support for any one of these alternative hypotheses requires further data from Petreşti-Arini as well as from the larger Transylvania area.

The lateral mesodermal divide: an epigenetic model of the origin of paired fins

By examining development at the level of tissues and processes, rather than focusing on gene expression, we have formulated a general hypothesis to explain the dorso-ventral and anterior-posterior placement of paired appendage initiation sites in vertebrates. According to our model, the number and position of paired appendages are due to a commonality of embryonic tissue environments determined by the global interactions involving the two separated layers (somatic and visceral) of lateral plate mesoderm along the dorso-ventral and anterior-posterior axes of the embryo. We identify this distribution of developmental conditions, as modulated by the separation/contact of the two LPM layers and their interactions with somitic mesoderm, ectoderm, and endoderm as a dynamic developmental entity which we have termed the lateral mesodermal divide (LMD). Where the divide results in a certain tissue environment, fin bud initiation can occur. According to our hypothesis, the influence of the developing gut suppresses limb initiation along the midgut region and the ventral body wall owing to an "endodermal predominance." From an evolutionary perspective, the lack of gut regionalization in agnathans reflects the ancestral absence of these conditions, and the elaboration of the gut together with the concomitant changes to the LMD in the gnathostomes could have led to the origin of paired fins.

Bone quality and bone strength: benefits of the bone-forming approach

The ability of bone to resist fracture depends on the intrinsic properties of the materials that comprise the bone matrix mineralization, the amount of bone (i.e. mass), and the spatial distribution of the bone mass (i.e. microarchitecture). Antiresorptive agents may prevent the decay of cancellous bone and cortical thinning, with no improvement of bone microstructure, leading to a partial correction of the principal bone quality defect in osteoporosis, the disruption of trabecular microarchitecture. Anabolic agents promote bone formation at both trabecular and endocortical surfaces, resulting in an increase of cancellous bone volume and cortical thickness. The improvement of cortical bone strength may be limited by an increase in cortical porosity. strontium ranelate improves trabecular network and cortical thickness that will contribute to anti-fracture efficacy at both vertebral and non-vertebral sites. The results of clinical and experimental studies are consistent with the mode of action of strontium involving dissociation between bone formation and resorption leading to a stimulation both trabecular and cortical bone formation without increasing cortical porosity.