bluuebunny/crossref_metadata_2025_split huggingface dataset:
An analysis of skeletal development in osteoblast-specific and chondrocyte-specific runt-related transcription factor-2 (Runx2) knockout mice
Authors: Takarada Takeshi , Hinoi Eiichi , Nakazato Ryota , Ochi Hiroki , Xu Cheng , Tsuchikane Azusa , Takeda Shu , Karsenty Gerard , Abe Takaya , Kiyonari Hiroshi , Yoneda Yukio 
Published: April 2013

ABSTRACT

Global gene deletion studies in mice and humans have established the pivotal role of runt related transcription factor-2 (Runx2) in both intramembranous and endochondral ossification processes during skeletogenesis. In this study, we for the first time generated mice carrying a conditional Runx2 allele with exon 4, which encodes the Runt domain, flanked by loxP sites. These mice were crossed with α1(I)-collagen-Cre or α1(II)-collagen-Cre transgenic mice to obtain osteoblast-specific or chondrocyte-specific Runx2 deficient mice, respectively. As seen in Runx2−/− mice, perinatal lethality was observed in α1(II)-Cre;Runx2flox/flox mice, but this was not the case in animals in which α1(I)-collagen-Cre was used to delete Runx2. When using double-staining with Alizarin red for mineralized matrix and Alcian blue for cartilaginous matrix, we observed previously that mineralization was totally absent at embryonic day 15.5 (E15.5) throughout the body in Runx2−/− mice, but was found in areas undergoing intramembranous ossification such as skull and clavicles in α1(II)-Cre;Runx2flox/flox mice. In newborn α1(II)-Cre;Runx2flox/flox mice, mineralization impairment was restricted to skeletal areas undergoing endochondral ossification including long bones and vertebrae. In contrast, no apparent skeletal abnormalities were seen in mutant embryo, newborn, and 3-week-old to 6-week old-mice in which Runx2 had been deleted with the α1(I)-collagen-Cre driver. These results suggest that Runx2 is absolutely required for endochondral ossification during embryonic and postnatal skeletogenesis, but that disrupting its expression in already committed osteoblasts as achieved here with the α1(I)-collagen-Cre driver does not affect overtly intramembranous and endochondral ossification. The Runx2 floxed allele established here is undoubtedly useful for investigating the role of Runx2 in particular cells. © 2013 American Society for Bone and Mineral Research.
An RNA-seq protocol to identify mRNA expression changes in mouse diaphyseal bone: Applications in mice with bone property alteringLrp5mutations
Authors: Ayturk Ugur M , Jacobsen Christina M , Christodoulou Danos C , Gorham Joshua , Seidman Jonathan G , Seidman Christine E , Robling Alexander G , Warman Matthew L 
Published: April 2013

ABSTRACT Loss-of-function and certain missense mutations in the Wnt coreceptor low-density lipoprotein receptor-related protein 5 (LRP5) significantly decrease or increase bone mass, respectively. These human skeletal phenotypes have been recapitulated in mice harboring Lrp5 knockout and knock-in mutations. We hypothesized that measuring mRNA expression in diaphyseal bone from mice with Lrp5 wild-type (Lrp5+/+), knockout (Lrp5–/–), and high bone mass (HBM)-causing (Lrp5p.A214V/+) knock-in alleles could identify genes and pathways that regulate or are regulated by LRP5 activity. We performed RNA-seq on pairs of tibial diaphyseal bones from four 16-week-old mice with each of the aforementioned genotypes. We then evaluated different methods for controlling for contaminating nonskeletal tissue (ie, blood, bone marrow, and skeletal muscle) in our data. These methods included predigestion of diaphyseal bone with collagenase and separate transcriptional profiling of blood, skeletal muscle, and bone marrow. We found that collagenase digestion reduced contamination, but also altered gene expression in the remaining cells. In contrast, in silico filtering of the diaphyseal bone RNA-seq data for highly expressed blood, skeletal muscle, and bone marrow transcripts significantly increased the correlation between RNA-seq data from an animal's right and left tibias and from animals with the same Lrp5 genotype. We conclude that reliable and reproducible RNA-seq data can be obtained from mouse diaphyseal bone and that lack of LRP5 has a more pronounced effect on gene expression than the HBM-causing LRP5 missense mutation. We identified 84 differentially expressed protein-coding transcripts between LRP5 “sufficient” (ie, Lrp5+/+ and Lrp5p.A214V/+) and “insufficient” (Lrp5–/–) diaphyseal bone, and far fewer differentially expressed genes between Lrp5p.A214V/+ and Lrp5+/+ diaphyseal bone. © 2013 American Society for Bone and Mineral Research.
Skeletal health in long-duration astronauts: Nature, assessment, and management recommendations from the NASA bone summit
Authors: Orwoll Eric S , Adler Robert A , Amin Shreyasee , Binkley Neil , Lewiecki E Michael , Petak Steven M , Shapses Sue A , Sinaki Mehrsheed , Watts Nelson B , Sibonga Jean D 
Published: April 2013

ABSTRACT

Concern about the risk of bone loss in astronauts as a result of prolonged exposure to microgravity prompted the National Aeronautics and Space Administration to convene a Bone Summit with a panel of experts at the Johnson Space Center to review the medical data and research evidence from astronauts who have had prolonged exposure to spaceflight. Data were reviewed from 35 astronauts who had served on spaceflight missions lasting between 120 and 180 days with attention focused on astronauts who (1) were repeat fliers on long-duration missions, (2) were users of an advanced resistive exercise device (ARED), (3) were scanned by quantitative computed tomography (QCT) at the hip, (4) had hip bone strength estimated by finite element modeling, or (5) had lost &gt;10% of areal bone mineral density (aBMD) at the hip or lumbar spine as measured by dual-energy X-ray absorptiometry (DXA). Because of the limitations of DXA in describing the effects of spaceflight on bone strength, the panel recommended that the U.S. space program use QCT and finite element modeling to further study the unique effects of spaceflight (and recovery) on bone health in order to better inform clinical decisions.
Electrochemical Behaviors of Glucose Oxidase Based on Biocatalytic Deposition of Gold Nanoparticles
Authors: He Yaping , Zheng Jianbin 
Published: April 2013

AbstractCoupling nanotechnology with biocatalysis, a highly sensitive glucose biosensor for the study of electrochemical behaviors of glucose oxidase (GOx) was proposed by using monkshoodvine root–bark like carbon (MLC) as the platform for the biocatalytic deposition of AuNPs. The biosensor showed a linear range from 0.25 to 130 μM with a detection limit of 0.1 μM (S/N = 3) towards glucose and sensitivity of 3010 μA/mM. K value was calculated to be 67.4 μM. Furthermore, the proposed AuNPs/GOx–MLC modified pyrolytic graphite electrode (AuNPs/GOx–MLC/PGE) achieved direct electron transfer of GOx. Γ* was calculated to be 2.79 × 10−11 mol/cm2 and ks was 1.79 s−1. It also showed a remarkable electrocatalysis towards glucose.
An Expedient Stereoselective Synthesis of Spirocyclopropyl Oxindoles from Indolin‐2‐One/N‐Protected Indolin‐2‐Ones and Bromonitroalkenes
Authors: Roy Suparna , Chen Kwunmin 
Published: April 2013

AbstractA direct and much simpler way was developed for the diastereoselective synthesis of spiro cyclopropan‐1,3′‐oxindoles from indolin‐2‐one/N‐protected indolin‐2‐ones and bromonitroalkene. The fused restrained cyclopropanes were obtained with high diastereomeric ratios (upto >99:1) and in reasonable to high isolated chemical yields (upto 94%).
Live cell fluorescence imaging for early expression and localization of RIB1 and RIB3 genes in Ashbya gossypii
Authors: Sengupta Sudeshna , Kaufmann Andreas , T. S. Chandra 
Published: April 2013

AbstractAshbya gossypii is a riboflavin overproducing filamentous fungus. RIB1 and RIB3 genes encode GTP‐cyclohydrolase II (GCH II) and DHBP synthase, respectively, the two rate limiting enzymes of the riboflavin biosynthetic pathway. The genes encoding yeast enhanced green fluorescent protein (yEGFP) and mCherry were fused with RIB1 and RIB3 genes, under their native promoters by PCR‐based gene tagging method for their early in vivo expression and cellular localization in A. gossypii. In the integrative transformants, the fusion proteins were expressed as cytoplasmic proteins from the germ bubble stage, in all the cells throughout the hypha. This was evident from the observation that mCherry fusion proteins were seen outside the vacuoles in the cytoplasm. The older matured cells of 14 h hyphae developed large vacuoles which showed green autofluorescence due to riboflavin. It is concluded that RIB1 and RIB3 genes are constitutively expressed in all the cells of this multicellular multinucleate fungus.
Identification of differentially expressed genes involved in laccase production in tropical white‐rot fungus Polyporus sp. PG15
Authors: Wang Jie , Zheng Xiaobing , Lin Shuoxin , Lin Junfang , Guo Liqiong , Chen Xiaoyang , Chen Qianting 
Published: April 2013

AbstractCarbon sources and copper ion are the main influencing factors on the production of fungal laccase. To investigate the regulation of carbon source and copper ion in laccase production on the molecular level in tropical white‐rot fungus PG15, a comparative analysis of gene expression patterns was performed by cDNA‐amplified fragment length polymorphism (AFLP) technique. Selective amplifications with 120 primer combinations allowed the identification of 92 differentially expressed transcript‐derived fragments (TDFs), ranging from 200 to 750 bp in size. The TDFs were from PG15 supplemented with different carbon sources and copper ion concentrations, majority of which downregulated laccase production. Twenty‐one fragments that matched the database were functionally annotated and analyzed according to the up‐ and downregulation patterns identified by cDNA‐AFLP. These fragments were probably involved in laccase production at the metabolism, signal transduction, transcription, or post‐translation levels. This study provides the first catalog of genes involved in laccase production, together with their putatively functional annotations. These data provide potential candidates for improving laccase production in fungi by marker‐assisted selection or genetic engineering.
PCR detection of ansA from marine bacteria and its sequence characteristics from Bacillus tequilensis NIOS4
Authors: Nayak Sagar , Porob Seema , Fernandes Areena , Meena Ram Murti , Ramaiah Nagappa 
Published: April 2013

AbstractAs many as 71 marine bacterial DNA extracts were PCR screened for L‐asparaginase (ansA), a key gene in anti‐cancer molecular‐searches. Over 62% (44) of them were positive for ansA gene. The positive cultures were from genera Bacillus and Staphylococcus. The ansA gene cloned from isolate NIOS4 belonging to recently described Bacillus tequilensis is 1099 bp in length with a 990 bp ORF coding for 329 amino acids. BLASTx analysis revealed this sequence to be 98% similar to earlier reported ansA sequence from B. subtilis (Accession no. NP390239.1). By comparing its deduced amino acid sequence with other bacterial asparaginase sequences six substitutions at positions 305(Thr), 313(Lys), 314(Leu), 315(Asp), 318(Arg), and 320(Gln) are observed. Key residues like Thr(12), Thr(85), Asp(86), Lys(156), and Phe(165) taking part in active‐site formation and imparting catalytic properties are conserved. The phylogenetic tree based of the ansA amino acid sequences revealed close relatedness of the NIOS4 ansA sequence with B. subtilis (Accession no. NP 390239.1). It's very close genetic resemblance to B. subtilis and conservation of certain key amino acid residues suggest it as a prospective candidate for evaluation and, production of L‐asparaginases.
In situ antioxidant and antimicrobial activities of naturally occurring caffeic acid, p‐coumaric acid and rutin, using food systems
Authors: Stojković Dejan , Petrović Jovana , Soković Marina , Glamočlija Jasmina , Kukić‐Marković Jelena , Petrović Silvana 
Published: April 2013

AbstractBackgroundThree pure compounds that naturally occur in plants were of particular interest to our study regarding the possibility of using them as food preservatives: p‐coumaric acid (found in peanuts, tomatoes, carrots, garlic, wine, vinegar, etc.), caffeic acid (found in argan oil, oats, wheat, rice and olive oil) and rutin (found in asparagus, citrus fruits, berries, apple, apricot, asparagus, beef and beer). In the following study we investigated in situ antioxidant and antimicrobial activities of three pure compounds, namely caffeic acid, p‐coumaric acid and rutin, naturally occurring in plants.ResultsTwo food systems were used in order to obtain information on how these compounds react in actual food systems rather than microbiological media. The results indicated good antioxidant activity in in situ food systems. For tested phenolic compounds it was further shown that they successively inhibited the development of the isolated food contaminant Staphylococcus aureus in chicken soup. Panelist found that organoleptic characteristics of chicken soup and pork meat improved after treatment with phenolics.ConclusionOur findings alone, along with the potential use of phenolic compounds that are widespread in nature, may imply their potential use as preservatives in the food industry. © 2013 Society of Chemical Industry
Structure analysis and antimutagenic activity of a novel salt‐soluble polysaccharide from Auricularia polytricha
Authors: Zhou Jie , Chen Yun , Xin Mengting , Luo Qinyao , Gu Jialu , Zhao Mengtian , Xu Xiaolan , Lu Xuanqi , Song Guanglei 
Published: April 2013

AbstractBackgroundAuricularia polytricha is known to be a highly nutritious foodstuff. We report here the purification, structure characterization and antimutagenic activity in vivo of a 0.9% NaCl solution‐soluble polysaccharide (SSP) from the mycelia of A. polytricha.ResultsAnalysis by high‐performance liquid chromatography coupled to a TSK‐G5000PWXL column and gel filtration chromatography on Sephacryl S‐400 HR indicated that SSP is homogeneous with an average molecular weight of about 9.30 × 105 Da. The structure of SSP was revealed by chemical methods, Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy. Results indicated that SSP is a glucan consisting of a1,3‐β‐glucan, 1,6‐α‐glucan, 1,4‐α‐glucan and 1,3‐α‐glucan backbone with a single 1,6‐α‐d‐glucopyranosyl side‐branching unit on every nine residues, on average, along the main chain. Atomic force microscopy indicates the presence of macromolecular species in morphology and shows a clear association of prolate particle. Meanwhile, SSP was found to significantly preventing micronuclei in polychromatic erythrocytes and reticulocytes of mice (P < 0.01).ConclusionThese results indicate that polysaccharide SSP from A. polytricha exhibits antimutagenic activity against the in vivo DNA‐damaging effect of the indirectly acting alkylating agent cyclophosphamide.© 2013 Society of Chemical Industry
Palliative surgery for head and neck cancer with extensive skin involvement
Authors: Jang David W. , Teng Marita S. , Ojo Bukola , Genden Eric M. 
Published: April 2013

Objectives/HypothesisTo evaluate the role of regional and free tissue transfer for the palliative management of head and neck cancer with extensive skin involvement.Study DesignCase Series.MethodsA retrospective review was performed of patients treated for head and neck cancer with involvement of the skin at the Mount Sinai Medical Center over a 5‐year period (2006–2010). Only patients with extensive skin involvement and unresectable tumors who underwent palliative resection and reconstruction were included in the review. Subjects were analyzed for age, gender, performance status, primary site, tumor histology, extent of invasion, type of reconstruction, hospital course, wound complications, adjuvant therapy, survival, and cause of death.ResultsTwenty‐five patients met the inclusion criteria for the review. Fourteen patients (56%) underwent regional flap reconstruction, and 11 patients (44%) underwent free flap reconstruction. The average length of stay was 7 days. Twenty‐four patients (96%) had a medically uncomplicated postoperative hospital course. Nineteen patients (76%) were treated with adjuvant palliative radiotherapy and/or chemotherapy. Long‐term follow‐up was achieved for 19 patients. The median follow up in this group was 9.5 months. Eleven of the 21 patients (52%) developed wound complications postoperatively. Eight of these were minor wound dehiscences, while three developed major wound complications. Four patients (16%) had distant metastasis at the time of surgery, and the median time to develop distant metastases after surgery was 6 months. Median survival time was 9.5 months. Twenty‐two patients (88%) were discharged in the care of their families with appropriate pain management and without the need for extensive wound care.ConclusionsFor unresectable tumors with extensive skin involvement, palliative resection and reconstruction is a reasonable treatment option. Although survival may not be affected, addressing the odor, bleeding, pain, and infection associated with skin involvement has the potential to improve a patient's quality of life.Level of Evidence4.
Ex Vivo Electromechanical Reshaping of Costal Cartilage in the New Zealand White Rabbit Model
Authors: Badran Karam , Manuel Cyrus , Waki Curtis , Protsenko Dmitry , Wong Brian J. F. 
Published: April 2013

Objectives/HypothesisDetermine the effective electromechanical reshaping (EMR) parameters for shape change and cell viability in the ex vivo rabbit costal cartilage model.Study DesignEx vivo animal study combined with computer modeling to guide electrode placement and polarity selection.MethodsRabbit costal cartilages were secured in a jig that approximated the shape of the rabbit auricle framework. Finite element modeling was used to select the initial electrode geometry, polarity, spacing, and estimate dosimetry parameters. Porcine costal cartilage was utilized to refine the selection of dosing parameters. Parametric analysis was performed to determine the effect of voltage and application time on tissue shape change. Next, rabbit rib cartilage was reshaped, varying voltage and application time to identify the lowest parameters to produce acceptable shape change mimicking native auricular cartilage. Acceptable qualitative shape change was determined on a five‐point Likert scale analyzed using one‐way general linear analysis of variance. Confocal microscopy with live/dead cell viability analysis determined the degree of injury and the distribution of live and dead cells.ResultsThe minimum acceptable deformation of rabbit costal cartilage was found at 4 V–3 minutes. Viability analysis of cartilage reshaped at 4 V–3 minutes demonstrates cell injury extending 2 mm away from each electrode with viable cells found between the electrodes.ConclusionsThe EMR parameters of 4 V–3 minutes demonstrates appropriate shape change producing grafts that resemble the native auricle and contains the viable cells adequate for clinical evaluation. The rabbit auricular reconstruction model using EMR is a feasible one.
The effect of steroids in combination With optic nerve decompression surgery in traumatic optic neuropathy
Authors: Ropposch Thorsten , Steger Bernhard , Meço Cem , Emesz Martin , Reitsamer Herbert , Rasp Gerd , Moser Gerhard 
Published: April 2013

Objectives/HypothesisTraumatic optic neuropathy (TON) is an important cause of severe visual impairment following blunt or penetrating head trauma. Treatment options include steroids, decompression, or both. Studies have failed to show a clear benefit for either steroids or surgery or a combined therapy. This study describes the visual outcome in surgically managed patients with or without steroid treatment.Study DesignRetrospective analysis.MethodsIn a retrospective chart review we included patients (n = 42) with TON who were managed with optic nerve decompression surgery with or without steroids. Comparison of initial and final visual acuity was the primary outcome parameter.ResultsIn 42 consecutive patients, steroids were used in 21 cases (50%). Ten patients received a high‐dose systemic regimen of more than 500 mg methylprednisolone initial dose, and 11 patients were treated with a lower dose. The overall visual improvement rate was 33%, no change was noted in 50%, and no outcome data were available in 17%. The visual improvement rate of patients treated with decompression surgery and steroids was lower (29%, 6 of 21) than those treated with surgery alone (53%, 8 of 15). Comparing both groups, additional steroids had no beneficial effect on the visual outcome (P = .97). There was no case with a deterioration of visual function due to therapy.ConclusionsIn patients with traumatic optic neuropathy who were treated with optic nerve decompression surgery, steroids had no beneficial effect on the visual outcome.Level of Evidence2c.
The effect of continuous positive airway pressure on middle ear pressure
Authors: Sivri Bekir , Sezen Ozan Seymen , Akbulut Sevtap , Coskuner Temel 
Published: April 2013

Objectives/HypothesisTo investigate the effect of continuous positive airway pressure (CPAP) on middle ear pressure in patients with obstructive sleep apnea syndrome (OSAS).Study DesignProspective clinical case‐control study.MethodsSeventy‐eight patients with moderate and severe OSAS (the mean apnea‐hypopnea indexes were 22.70 ± 3.59 and 48.59 ± 12.50, respectively) using CPAP (study group) and 60 patients with no sleep apnea syndrome (control group) were included in the study. Forty‐two of the patients were female and 96 were male. The mean age in the study group was 47.38 ± 6.23, whereas it was 46.10 ± 11.96 in the control group (P > .05). The middle ear pressure of all study subjects was evaluated with tympanometry at the beginning of the study and 6 months later. Tympanometric measurements of both groups were compared. Furthermore, the same data for the study group were also evaluated according to CPAP pressure levels.ResultsThe middle ear peak pressure values of the patients in the study group were significantly increased from −63.04 ± 55.82 daPa to −39.6 ± 27.72 daPa after 6 months (P < .01). The middle ear pressure in the control group was found to be −13.26 ± 22.60 daPa at the beginning of the study and −13.60 ± 38.82 daPa after 6 months (P > .05). The mean middle ear pressure level was significantly higher in patients using CPAP at 12 to 14 cm H2O pressure than in those using CPAP at 8 to 10 cm H2O pressure (P < .05).ConclusionsThere was a significant increase in the middle ear pressure of patients using CPAP regularly for 6 months. This increase was proportional to the pressure level of the CPAP device.Level of Evidence3b.