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Pulmonary hypertension (PH) is a fatal disorder characterized by pulmonary vascular remodeling and obstruction. The phosphodiesterase 4 (PDE4) family hydrolyzes cyclic AMP (cAMP) and is comprised of four subtypes (PDE4A–D). Previous studies have shown the beneficial effects of pan-PDE4 inhibitors in rodent PH; however, this class of drugs is associated with side effects owing to the broad inhibition of all four PDE4 isozymes. Here, we demonstrate that PDE4B is the predominant PDE isozyme in lungs and that it was upregulated in rodent and human PH lung tissues. We also confirmed that PDE4B is mainly expressed in the lung endothelial cells (ECs). Evaluation of PH in Pde4b wild type and knockout mice confirmed that Pde4b is important for the vascular remodeling associated with PH. In vivo EC lineage tracing demonstrated that Pde4b induces PH development by driving endothelial-to-mesenchymal transition (EndMT), and mechanistic studies showed that Pde4b regulates EndMT by antagonizing the cAMP-dependent PKA–CREB–BMPRII axis. Finally, treating PH rats with a PDE4B-specific inhibitor validated that PDE4B inhibition has a significant pharmacological effect in the alleviation of PH. Collectively, our findings indicate a critical role for PDE4B in EndMT and PH, prompting further studies of PDE4B-specific inhibitors as a therapeutic strategy for PH. Pan-PDE4 inhibitors attenuate rat pulmonary hypertension (PH), while its applications show serious gastrointestinal side-effects. PDE4B-specific inhibitor may be an advisable therapeutic strategy that can alleviate PH by suppressing EndMT with less side-effects. [Display omitted] [ABSTRACT FROM AUTHOR]

Background: Kalkitoxin (KT) is an active lipopeptide isolated from the cyanobacterium Lyngbya majuscula found in the bed of the coral reef. Although KT suppresses cell division and inflammation, KT's mechanism of action in vascular smooth muscle cells (VSMCs) is unidentified. Therefore, our main aim was to investigate the impact of KT on vascular calcification for the treatment of cardiovascular disease. Objectives: Using diverse calcification media, we studied the effect of KT on VSMC calcification and the underlying mechanism of this effect. Methods: VSMC was isolated from the 6 weeks ICR mice. Then VSMCs were treated with different concentrations of KT to check the cell viability. Alizarin red and von Kossa staining were carried out to examine the calcium deposition on VSMC. Thoracic aorta of 6 weeks mice were taken and treated with different concentrations of KT, and H and E staining was performed. Real-time polymerase chain reaction and western blot were performed to examine KT's effect on VSMC mineralization. Calcium deposition on VSMC was examined with a calcium deposition quantification kit. Results: Calcium deposition, Alizarin red, and von Kossa staining revealed that KT reduced inorganic phosphate-induced calcification phenotypes. KT also reduced Ca++-induced calcification by inhibiting genes that regulate osteoblast differentiation, such as runt- related transcription factor 2 (RUNX-2), SMAD family member 4, osterix, collagen 1α, and osteopontin. Also, KT repressed Ca2+-induced bone morphogenetic protein 2, RUNX-2, collagen 1α, osteoprotegerin, and smooth muscle actin protein expression. Likewise, Alizarin red and von Kossa staining showed that KT markedly decreased the calcification of ex vivo ring formation in the mouse thoracic aorta. Conclusions: This experiment demonstrated that KT decreases vascular calcification and may be developed as a new therapeutic treatment for vascular calcification and arteriosclerosis. [ABSTRACT FROM AUTHOR]

Protein adsorption preferentially occurs and significantly affects the physicochemical reactions once the biodegradable magnesium alloys as bone replacements have been implanted. To date, interactions mechanisms between Mg implants and proteins remain unclear at a molecular level. Thereby, a combination of molecular dynamic (MD) simulations and experimental exploration is used to investigate the adsorption behavior and conformational change of bovine serum albumin (BSA), a representative protein of blood plasma, upon the surface of micro-arc oxidation (MAO) coated Mg alloy AZ31. The influences of absorbed proteins on the cytocompatibility of MAO coating are evaluated by virtue of cytotoxicity assay. Results indicate that the negatively charged O atoms (BSA) exhibit strong interaction with Mg2+ ions of Mg(OH) 2 , revealing that BSA molecules are ionically adsorbed on the AZ31 surface. Interestingly, MD simulation reveals that MAO coating demonstrates superior ability to capture BSA molecules during the process of adsorption owing to strong electric attraction between the negatively charged O atoms in BSA molecules with Mg atoms of MgO in MAO coating. Moreover, the α -helix part of absorbed BSA molecules on AZ31 substrate and MAO coating markedly decreases with an increase in β -sheet, β -turn and unordered contents, which is attributed to the reduction in the number of hydrogen bonds in BSA molecules. Furthermore, the adsorbed BSA molecules improve the cytocompatibility of MAO coating since the positively charged -NH 3 + group and β -sheet content of absorbed BSA molecules mediate the cell adhesion by interacting with the negatively charged cell membrane. [ABSTRACT FROM AUTHOR]

As one of the most common pathological changes in trauma and surgery practice, intestinal ischemia-reperfusion (I/R) injury is regarded as a major precipitating factor in the occurrence and development of fatal diseases. BRCA1--BRCA2-containing complex subunit 36 (BRCC36), a deubiquitinase, has been proved important in a variety of pathophysiological processes such as DNA repair, cell cycle regulation, tumorigenesis, and inflammatory response. However, the effect of BRCC36 on intestinal mucosal barrier injury after I/R has not been fully elucidated. Our research found that BRCC36 aggravated intestinal mucosal barrier injury caused by bone morphogenetic protein 2 after I/R by downregulating peroxisome proliferator-activated receptor-γ (PPAR γ) signaling. These results suggested that BRCC36/PPAR γ axis might serve as a potential therapeutic target for preventing intestinal mucosal barrier injury after I/R. [ABSTRACT FROM AUTHOR]

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Background: Alfalfa(Medicago sativa L.) is a perennial plant, which is high in nutritional value and resistant to environmental conditions, and it is one of most frequently preferred feed crop among the leguminous family. In this study, it was aimed to determine the genetic diversity of some alfalfa ecotypes and their varieties by DNA, protein, nucleus, and chromosome counts. The genetic distance between the populations of control (M. truncatula), five different cultivars (Alsancak, Bilensoy, Iside, Plato, Bilensoy82), and three different ecotypes (Erzurum, Muş, and Konya) was investigated by cytogenetic analysis, flow cytometry, simple sequence repeats (SSR), and SDS PAGE techniques. Results: Cytogenetic analysis of these tested plants has verified the existence of expected levels such as diploid, triploid, and tetraploid as well as aneuploid (2n = 4x = 30) plants. Flow cytometry analysis have displayed that all of tested plants were tetraploid, whereas cytological analysis had either diploid, triploid, or tetraploid. Genetic diversity dendrogram was created using Erzurum, Muş, Konya, Bilensoy82, Alsancak, and Plato varieties. The Iside and Bilensoy were found to be morphogenetic in relationship. Our control plant, M. truncatula, did not have a similarity relationship with other ecotypes and cultivars. The total numbers of protein bands differed among tested plants from 140 kDA to 25 kDa. Conclusions: This paper first reports on the genetic variation of Turkish alfalfa plants by using detailed analysis techniques. This work provides important findings for the classification, conservation, and innovation of alfalfa germplasm resources. [ABSTRACT FROM AUTHOR]

Amorphous Ca-phosphate (ACP) particles stabilized by inorganic polyphosphate (polyP) were prepared by co-precipitation of calcium and phosphate in the presence of polyP (15% [w/w]). These hybrid nanoparticles showed no signs of crystallinity according to X-ray diffraction analysis, in contrast to the particles obtained at a lower (5% [w/w]) polyP concentration or to hydroxyapatite. The ACP/15% polyP particles proved to be a suitable matrix for cell growth and attachment and showed pronounced osteoblastic and vasculogenic activity in vitro. They strongly stimulated mineralization of the human osteosarcoma cell line SaOS-2, as well as cell migration/microvascularization, as demonstrated in the scratch assay and the in vitro angiogenesis tube forming assay. The possible involvement of an ATP gradient, generated by polyP during tube formation of human umbilical vein endothelial cells, was confirmed by ATP-depletion experiments. In order to assess the morphogenetic activity of the hybrid particles in vivo , experiments in rabbits using the calvarial bone defect model were performed. The particles were encapsulated in poly(d , l -lactide- co -glycolide) microspheres. In contrast, to crystalline Ca-phosphate (containing only 5% [w/w] polyP) or to crystalline β-tricalcium phosphate, amorphous ACP/15% polyP particles caused pronounced osteoinductive activity already after a six-week healing period. The synthesis of new bone tissue was accompanied by an intense vascularization and an increased expression of mineralization/vascularization marker genes. The data show that amorphous polyP-stabilized ACP, which combines osteoinductive activity with the ability to act as a precursor of hydroxyapatite formation both in vitro and in vivo , is a promising material for bone regeneration. Image, graphical abstract [ABSTRACT FROM AUTHOR]

While human bone morphogenetic protein-2 (BMP-2) is a promising growth factor for bone regeneration, a major challenge in biomedical applications is finding an optimal carrier for its delivery at the site of injury. Because of their natural affinities for growth factors (including BMP-2) as well as their role in instructing cell function, cultured cell-derived extracellular matrices (ECM) are of special interest. We hereby hypothesized that a "bony matrix" containing mineralized, osteogenic ECM is a potential efficacious carrier of BMP-2 for promoting bone formation and, therefore, compared the efficacy of the decellularized ECM derived from osteogenic-differentiated human mesenchymal stem cells (hMSCs) to the one obtained from ECM from undifferentiated hMSCs. Our results provided evidence that both ECMs can bind BMP-2 and promote bone formation when implanted ectopically in mice. The osteoinductive potential of BMP-2, however, was greater when loaded within an osteogenic MSC-derived ECM; this outcome was correlated with higher sequestration capacity of BMP-2 over time in vivo. Interestingly, although the BMP-2 mainly bound onto the mineral crystals contained within the osteogenic MSC derived-ECM, these mineral components were not involved in the observed higher osteoinductivity, suggesting that the organic components were the critical components for the matrix efficacy as BMP-2 carrier. Image, graphical abstract [ABSTRACT FROM AUTHOR]

The paper presents issues regarding food safety and food contamination sources with polycyclic aromatic hydrocarbons. The main food processing methods that contribute to the contamination with polycyclic aromatic hydrocarbons are smoking, frying, baking, obtaining grills, using polymer packaging. Benzopyrene is the most carcinogenic polycyclic aromatic hydrocarbon, formed as a result of food preparation processes with starch, at temperatures of 370-650 degrees Celsius. Biological monitoring of PAH exposure is important due to the toxicity and reactivity of their metabolites (epoxides and dihydrodiols), which can react with cellular proteins and as a result, DNA is not synthesized and genetic mutations, malformations, tumors and cancer develop. [ABSTRACT FROM AUTHOR]

The modulation of cell behavior during culture is one of the most important aspects of bone tissue engineering because of the necessity for a complex mechanical and biochemical environment. This study aimed to improve the physicochemical properties of chitosan/multi-phase calcium phosphate (MCaP) scaffolds using an optimized mixture design experiment and evaluate the effect of biofunctionalization of the obtained scaffolds with the bone morphogenetic protein BMP-2 on stem cell behavior. The present study evaluated the compressive strength, elastic modulus, porosity, pore diameter, and degradation in simulated body fluids and integrated these responses using desirability. The properties of the scaffolds with the best desirability (18.4% of MCaP) were: compressive strength of 23 kPa, elastic modulus of 430 kPa, pore diameter of 163 μ m, porosity of 92%, and degradation of 20% after 21 days. Proliferation and differentiation experiments were conducted using dental pulp stem cells after grafting BMP-2 onto scaffolds via the carbodiimide route. These experiments showed that MCaP promoted cell proliferation and increased alkaline phosphatase activity, whereas BMP-2 enhanced cell differentiation. This study demonstrates that optimizing the composition of a mixture of chitosan and MCaP improves the physicochemical and biological properties of scaffolds, indicating that this solution is viable for application in bone tissue engineering. [Display omitted] [ABSTRACT FROM AUTHOR]

Induced pluripotent stem (iPS) cells could be induced into ameloblast-like cells by ameloblasts serum-free conditioned medium (ASF-CM), and bone morphogenetic proteins (BMPs) might be essential during the regulation of this process. The present study investigates the signal transduction that regulates the ameloblastic differentiation of iPS cells induced by ASF-CM. Mouse iPS cells were characterized and then cultured for 14 days in epithelial cell medium (control) or ASF-CM. Bone morphogenetic protein receptor II (BMPR-II) siRNA, inhibitor of Smad1/5 phosphorylation activated by activin receptor-like kinase (ALK) receptors, and inhibitors of mitogen-activated protein kinases (MAPKs) phosphorylation were used to treat the iPS cells in combination with ASF-CM. Real-time PCR, western blotting, and immunofluorescent staining were used to evaluate the expressions of ameloblast markers ameloblastin, enamelin, and cytokeratin-14. BMPR-II gene and protein levels increased markedly in ASF-CMtreated iPS cells compared with the controls, while the mRNA levels of Bmpr-Ia and Bmpr-Ib were similar between the ASF-CM and control groups. ASF-CM stimulation significantly increased the gene and protein expression of ameloblastin, enamelin and cytokeratin-14, and phosphorylated SMAD1/5, p38 MAPK, and ERK1/2 MAPK compared with the controls. Knockdown of BMPR-II and inhibition of Smad1/5 phosphorylation both could significantly reverse the increased expression of ameloblastin, enamelin, and cytokeratin-14 induced by ASF-CM, while neither inhibition of p38 nor ERK1/2 phosphorylation had significant reversing effects. We conclude that smad1/5 signaling transduction, activated by ALK receptors, regulates the ameloblastic differentiation of iPS cells induced by ameloblast-conditioned medium. [ABSTRACT FROM AUTHOR]

Supplemental Digital Content is Available in the Text. Purpose: To investigate clinical factors and cellular responses of in situ human alveolar bone–derived mesenchymal stromal cells involved in early periimplant marginal bone loss. Materials and Methods: Thirty-seven completely or partially edentulous patients were enrolled in this study. Periapical radiographs were taken at the time of implant surgery, at 3-month follow-up, and at 1-year follow-up. Univariate analysis and multiple logistic regression were performed to investigate the associations between marginal bone loss and study variables. The mRNA expression levels of 21 bone-remodeling– and tissue-healing–associated genes were analyzed by subgroup. Results: Thirty-one patients with 98 implants were followed. The incidence and mean amount of bone loss were higher for overdentures than for other prosthesis and higher for the maxilla than for the mandible. The bone loss group showed lower mRNA expression levels of runt-related transcription factor-2, bone morphogenetic protein-2, and peroxisome proliferator–activated receptor gamma-2 and higher receptor activator of NKκB ligand/osteoprotegerin (RANKL/OPG) ratio. Conclusion: Within the limitations of the study, certain genes involved in bone remodeling (runt-related transcription factor-2 [Runx-2], bone morphogenetic protein-2 [BMP-2], and peroxisome proliferator–activated receptor gamma-2 [PPARγ-2]) and RANKL/OPG are correlated with early periimplant bone loss, with the type of suprastructure and the involved jaw being significant clinical factors. [ABSTRACT FROM AUTHOR]

Mitogen-Activated Protein (MAP) Kinase pathway involves several oncogenic genes which can serve as potential targets for therapy. Therefore, aim of the present study is to analyze mutations in the MAP Kinase pathway in pulmonary adenocarcinoma (ADCA) of Indian patients along with clinico-pathologic correlation and determination of the survival status in patients receiving therapy. Blocks and slides of 125 pulmonary ADCA of last 5 years were retrieved. Histo-morphology and tumor content were determined. EGFR, KRAS, BRAF and MEK1 genes were analyzed using Sanger sequencing and Real-time polymerase chain reaction (PCR). Clinico-pathologic correlation and survival analysis were performed. Fifty-eight (46.4%) patients harbored genetic mutations of which 49 had single somatic mutations, 5 had multiple exonic and 4 showed coexisting EGFR and KRAS mutations. EGFR mutations were seen in 24.8%, KRAS in 19.2% and BRAF (non-V600E) in 2.4% cases. There was no difference in progression-free survival of wild- type/single mutations when compared with multiple/ coexisting mutations (P = 0.09). However, the P value may indicate borderline correlation. To conclude, EGFR and KRAS mutations may coexist in the same patient in lung ADCA. Multiple exonic mutations of KRAS gene formed substantial percentage of our cohort, requiring further exploration. Lung ADCA harbouring BRAF mutations are commonly non-V600E. Testing of all major genetic driver mutations of lung ADCA irrespective of histology and other demographic characteristics is necessary. [ABSTRACT FROM AUTHOR]

Barrier membranes are essential biomaterials for guided bone regeneration. Due to different origin and structure of barrier membranes, singular mechanical properties and clinical behaviors can be expected. It is important to understand the physic and chemical properties of barrier membranes to select the needed biomaterial for each clinical situation. To date, no study has evaluated and compared the physicochemical properties of various families of barrier membranes. The aim of this study is to evaluate the physicochemical properties of various barrier membranes. Fifteen membranes of different origin were tested in this study. Membranes were divided into biological or synthetic origin and grouped in natural allogenic collagen, natural xenogenic collagen, cross-linked collagen and synthetic membranes. Physicochemical properties were evaluated in terms of tension, stiffness, absorption ability, pH and wettability. For the tension tests, all membranes showed similar low tension and low stiffness, especially after a 4-min hydration, except for bone laminas that showed a greater stiffness particularly in a dry status. Regarding wettability and hydration of the barrier membranes, porcine origin membranes had greater hydration; wettability was also superior in porcine derived barrier membranes and showed a faster absorption of the drop on the rough surfaces. All membranes had a stable pH, having the synthetic membranes the most stable pH when compared to physiologic. The wide variety of barrier membranes opens a debate in which the practitioner should select the adequate barrier membrane for each clinical situation. Different materials show singular potentials depending on their tissue origin making them suitable for specific clinical indications. More studies regarding adsorption, integration and degradation of barrier membranes are needed to understand their behavior. Image 1 • There are a wide variety of barrier membranes with different origin and composition. • Barrier membranes differ on their physicochemical properties under different conditions. • Alloplastic barrier membranes shared similar physicochemical properties. • There are a wide variety of barrier membranes that can fulfill different clinical necessities. • No ideal barrier membrane exists in terms of physicochemical properties. [ABSTRACT FROM AUTHOR]

To observe the effect of high positive acceleration (+Gz) environment on dental implant osseointegration in a rabbit model and to investigate its mechanism. Forty-eight New Zealand white rabbits were randomly divided into 6 groups. The rabbit's mandibular incisors were extracted and 1 implant was placed in each socket immediately. After 1 week of rest, the rabbits were exposed to a high +Gz environment, 3 times a week. The rabbits were sacrificed at 3 weeks (2 weeks +Gz exposure), 5 weeks (4 weeks +Gz exposure), and 12 weeks (4 weeks +Gz exposure and 7 weeks normal environment) after surgery, respectively. Specimens were harvested for micro-CT scanning, histological analysis, and real-time polymerase chain reaction examination. Compared with those in the control group, the mRNA expression levels of bone morphogenetic protein-2 (BMP-2), osteopontin (OPN), and transforming growth factor-β1 (TGF-β1) were significantly lower (P < 0.05), while the mRNA expression level of receptor activator of nuclear factor k B ligand (RANKL) and the RANKL/osteoprotegerin (OPG) ratio were significantly higher (P < 0.05) at 3 weeks; values of bone volume fraction, trabecular number, bone-implant contact (BIC), and TGF-β1 and OPG mRNA expression levels were significantly lower (P < 0.05), and the value of trabecular separation, RANKL mRNA expression level and RANKL/OPG ratio were significantly higher (P < 0.05) at 5 weeks; and the value of BIC was still significantly lower (P < 0.05) at 12 weeks in the experimental group. Early exposure to the high +Gz environment after implant surgery might have an adverse effect on osseointegration, and its mechanism could be related to the inhibition of osteoblast activity and promotion of osteoclast activity. [ABSTRACT FROM AUTHOR]

Sclerostin is a protein which is involved in bone metabolism and probably also in vessel wall function. This prospective observational cohort study evaluated the prognostic significance of sclerostin in hemodialysis (HD) patients. In total, 106 HD patients and 25 healthy controls participated in the study. HD patients were prospectively followed up for five years. Sclerostin was measured in serum using standard ELISA kits by Biomedica. Sclerostin concentrations in serum were higher in HD patients compared to the controls (89.2±40.3 pmol/l vs. 32.8±13.0 pmol/l, p<0.001). Sclerostin levels were significant for cardiovascular mortality but not for overall mortality and mortality due to infection. A higher cardiovascular risk was connected to sclerostin concentrations above the median (>84 pmol/l), HR (95 % CI): 2.577 (1.0002-10.207), p=0.04. When sclerostin was evaluated together with residual diuresis in Kaplan-Meier analysis the worst prognosis due to cardiovascular events was observed in the group with high sclerostin and zero residual diuresis compared to all other patients (p=0.007). In summary, serum sclerostin levels in HD patients were increased when compared to healthy subjects. High sclerostin levels were demonstrated as a risk factor for cardiovascular mortality. Further studies are required to clarify the pathophysiological mechanisms of sclerostin action in patients with renal failure before therapeutic measures can be established. [ABSTRACT FROM AUTHOR]

Summary Background Rare genetic variants cause pulmonary arterial hypertension, but the contribution of common genetic variation to disease risk and natural history is poorly characterised. We tested for genome-wide association for pulmonary arterial hypertension in large international cohorts and assessed the contribution of associated regions to outcomes. Methods We did two separate genome-wide association studies (GWAS) and a meta-analysis of pulmonary arterial hypertension. These GWAS used data from four international case-control studies across 11 744 individuals with European ancestry (including 2085 patients). One GWAS used genotypes from 5895 whole-genome sequences and the other GWAS used genotyping array data from an additional 5849 individuals. Cross-validation of loci reaching genome-wide significance was sought by meta-analysis. Conditional analysis corrected for the most significant variants at each locus was used to resolve signals for multiple associations. We functionally annotated associated variants and tested associations with duration of survival. All-cause mortality was the primary endpoint in survival analyses. Findings A locus near SOX17 (rs10103692, odds ratio 1·80 [95% CI 1·55–2·08], p=5·13 × 10–15) and a second locus in HLA-DPA1 and HLA-DPB1 (collectively referred to as HLA-DPA1/DPB1 here; rs2856830, 1·56 [1·42–1·71], p=7·65 × 10–20) within the class II MHC region were associated with pulmonary arterial hypertension. The SOX17 locus had two independent signals associated with pulmonary arterial hypertension (rs13266183, 1·36 [1·25–1·48], p=1·69 × 10–12; and rs10103692). Functional and epigenomic data indicate that the risk variants near SOX17 alter gene regulation via an enhancer active in endothelial cells. Pulmonary arterial hypertension risk variants determined haplotype-specific enhancer activity, and CRISPR-mediated inhibition of the enhancer reduced SOX17 expression. The HLA-DPA1/DPB1 rs2856830 genotype was strongly associated with survival. Median survival from diagnosis in patients with pulmonary arterial hypertension with the C/C homozygous genotype was double (13·50 years [95% CI 12·07 to >13·50]) that of those with the T/T genotype (6·97 years [6·02–8·05]), despite similar baseline disease severity. Interpretation This is the first study to report that common genetic variation at loci in an enhancer near SOX17 and in HLA-DPA1/DPB1 is associated with pulmonary arterial hypertension. Impairment of SOX17 function might be more common in pulmonary arterial hypertension than suggested by rare mutations in SOX17. Further studies are needed to confirm the association between HLA typing or rs2856830 genotyping and survival, and to determine whether HLA typing or rs2856830 genotyping improves risk stratification in clinical practice or trials. Funding UK NIHR, BHF, UK MRC, Dinosaur Trust, NIH/NHLBI, ERS, EMBO, Wellcome Trust, EU, AHA, ACClinPharm, Netherlands CVRI, Dutch Heart Foundation, Dutch Federation of UMC, Netherlands OHRD and RNAS, German DFG, German BMBF, APH Paris, INSERM, Université Paris-Sud, and French ANR. [ABSTRACT FROM AUTHOR]

Abstract The swimming crab (Portunus trituberculatus , Portunus) can tolerate low salinity, but the mechanism of its varied salinity adaptation at the molecular level remains unclear. In this study, we prepared four mRNA and microRNA (miRNA) libraries using the gills collected from four salinity groups and performed RNA-sequencing (RNA-Seq) to identify the genes related to the low salinity. We set 25 ppt as the control group. A total of 659 genes were differentially expressed in at least one of the six comparison groups (25 ppt vs. 20 ppt, 25 ppt vs. 15 ppt, 25 ppt vs. 10 ppt, 20 ppt vs. 15 ppt, 20 ppt vs. 10 ppt and 15 ppt vs. 10 ppt). A total of 15 and 9 unigenes were downregulated and upregulated under low salinity compared with that in 25 ppt, respectively. Six genes, namely, aminopeptidase , centromere protein , cytochrome b5 reductase , bone morphogenetic protein , and two carbonic anhydrases , were selected for verification through quantitative real-time PCR. The results were consistent with the RNA-Seq results. Furthermore, 95 conserved miRNAs and 16 novel miRNAs were differentially expressed in at least one of the six comparison groups. Analysis of the miRNA–mRNA interaction showed that miR-2 and miR-317 regulated >50 mRNA targets. In addition, let-7c was downregulated in all groups under low salinity compared with that in the control group. This study helped elucidate the adaptation mechanism of the swimming crab in low-saline environments. Graphical abstract Unlabelled Image [ABSTRACT FROM AUTHOR]

Implant-related osteomyelitis harms patients' health and lives. Non-invasive treatments triggered by external physical stimuli have been brought into focus in antibiosis. Herein, a therapeutic strategy through coupling piezoelectricity and pyroelectricity of BaTiO 3 nanotubes (BNT) by synchronous treatments of ultrasound (US) and near-infrared light (NIR) is applied to kill bacteria in an osteomyelitis model. NIR or US treatment reduces spontaneous polarization of BNT, inducing pyroelectric/piezoelectric field and energy band tilting. Compared with single treatment, their synergistic effect further enhances charge transfer and electron-hole pair separation, improving the pyro-piezoelectric catalysis and thus the yield of reactive oxygen species (ROS); meanwhile, the tilting of energy bands induces an irreversible conductive circuit from bacteria to BNT, resulting in membrane electron loss and the disrupted bacterial metabolic balance. As a result, 99.8% of bacteria are killed under the pyro-piezoelectric therapy of BNT. Additionally, BNT with nanotubular structure acts as nanocarriers to load and release bone morphogenetic protein-2 (BMP-2), improving bone regeneration obviously after antibacterial therapy. This study provides a promising pyro-piezoelectric therapy by BMP-2 loaded BaTiO 3 nanotubes for antibiosis and bone regeneration in treating osteomyelitis. It also brings new insight for developing pyro-piezoelectric sensors to deal with tissue infection. [Display omitted] • The pyro-piezoelectricity of BaTiO 3 nanotubes is applied to treat osteomyelitis. • BaTiO 3 nanotubes generate more ROS by the synergy of pyro- and piezo-electricity. • BaTiO 3 nanotubes plunder bacterial electrons due to the further tilted energy band. • BaTiO 3 nanotubes load and release BMP-2, improving osteogenesis after sterilization. [ABSTRACT FROM AUTHOR]

Localization of recombinant human bone morphogenetic protein-2 (rhBMP-2) with continuous and effective osteogenic stimulation is still a great challenge in the field of bone regeneration. To achieve this aim, rhBMP-2 was tethered on chondroitin sulfate (CS)-functionalized calcium phosphate cement (CPC) scaffolds through specific noncovalent interactions. CS, one of the core glycosaminoglycans, was covalently conjugated onto CPC scaffolds with the assistance of polydopamine (PDA) and further immobilized rhBMP-2 in a biomimetic form. The CPC-PDA-CS scaffolds not only controlled the release kinetics and presentation state of rhBMP-2 but also effectively increased the expression levels of bone morphogenetic protein receptors (BMPRs) and enhanced the recognitions of the remaining rhBMP-2 to BMPRs. Strikingly, the rhBMP-2-loaded CPC-PDA-CS significantly promoted the cellular surface translocation of BMPRs (especially BMPR-IA). In vivo studies demonstrated that, compared with the rhBMP-2 upon CPC and CPC-PDA, the rhBMP-2 upon CPC-PDA-CS exhibited sustained release and induced high quality and more ectopic bone formation. Collectively, these results suggest that rhBMP-2 can be localized within CS-functionalized CPC scaffolds and exert continuous, long-term, and effective osteogenic stimulation. Thus, this work could provide new avenues in mimicking bone extracellular matrix microenvironment and localizing growth factor activity for enhanced bone regeneration. Statement of Significance A bioinspired chondroitin sulfate (CS)-functionalized calcium phosphate cement (CPC) platform was developed to tether recombinant human bone morphogenetic protein-2 (rhBMP-2), which could exhibit continuous, long-term, and effective osteogenic stimulation in bone tissue engineering. Compared with rhBMP-2-loaded CPC, the rhBMP-2-loaded CPC-polydopamine-CS scaffolds induced higher expression of bone morphogenetic protein receptors (BMPRs), greater cellular surface translocation of bone morphogenetic protein receptor-IA, higher binding affinity of BMPRs/rhBMP-2, and thus higher activation of the drosophila gene mothers against decapentaplegic protein-1/5/8 (Smad1/5/8) and extracellular-regulated protein kinases-1/2 (ERK1/2) signaling. This work can provide new guidelines for the design of BMP-2-based bioactive materials for bone regeneration. [ABSTRACT FROM AUTHOR]

The role and mechanisms of integrated stress response inhibitor (ISRIB) on silicosis are still not well defined. In the present study, the effects of ISRIB on cellular senescence and pulmonary fibrosis in silicosis were evaluated by RNA sequencing, micro-computed tomography, pulmonary function assessment, histological examination, and Western blot analysis. The results showed that ISRIB significantly reduced the degree of pulmonary fibrosis in mice with silicosis and reduced the expression of type I collagen, fibronectin, α-smooth muscle actin, and transforming growth factor-β1. Both in vivo and in vitro results showed that ISRIB reversed the expression of senescence-related factors β-galactosidase, phosphor-ataxia telangiectasia mutated, phosphor-ataxia telangiectasia and Rad3-related protein, p-p53, p21, p16, and plasminogen activator inhibitor type 1. The aforementioned results were consistent with the sequencing results. These findings implied that ISRIB might reduce the degree of pulmonary fibrosis in mice with silicosis by inhibiting the cellular senescence of alveolar epithelial cell type II. • ISRIB alleviates pulmonary fibrosis induced by silica in mice. • ISRIB inhibits cellular senescence of type II pulmonary epithelial cells. • A crosstalk exists between cellular senescence and the PERK signaling pathway. [ABSTRACT FROM AUTHOR]

It has been reported that particulate matter with an aerodynamic diameter of <2.5 µm (PM2.5) could induce epithelial–mesenchymal transition (EMT)- and extracellular matrix (ECM)-related pulmonary fibrosis (PF). The transcription factor Nrf2 alleviated PM2.5-induced PF by antagonizing oxidative stress. The N6-methyladenosine (m6A) modification plays a significant role in the stress response. However, the effect of m6A modification on the mechanisms of Nrf2-mediated defense against PM2.5-induced PF remained unknown. Here, we explored the role and the underlying molecular mechanisms of m6A methylation of Nrf2 mRNA in PM2.5-induced PF. We established filtered air (FA), unfiltered air (UA), and concentrated PM2.5 air (CA) group mice model and 0, 50, and 100 μg/mL PM2.5-treated 16HBE cell models. The extent of lung fibrosis in mice and fibrosis indicators were detected by histopathological analysis, immunohistochemical staining and western blotting. The molecular mechanism of m6A-modified Nrf2 was demonstrated by m6A-methylated RNA immunoprecipitation (MeRIP), RNA immunoprecipitation (RIP), qRT-PCR and T3 ligase-based PCR. Our data showed that PM2.5 exposure for 16 weeks could induce pulmonary fibrosis and activate Nrf2 signaling pathway. m6A methyltransferase METTL3 was upregulated after PM2.5 treatment in vivo and in vitro. Moreover, METTL3 mediated m6A modification of Nrf2 mRNA and promoted Nrf2 translation in mice and 16HBE cells after PM2.5 exposure. Mechanistically, three m6A-modified sites (1317, 1376 and 935; numbered relative to the first nucleotide of 3'UTR) of Nrf2 mRNA were identified in PM2.5-treatment 16HBE cells. Furthermore, the m6A binding proteins YTHDF1/IGF2BP1 promoted Nrf2 translation by binding to m6A residues of Nrf2 mRNA. Our results revealed the mechanism of m6A mediated Nrf2 signaling pathway against oxidative stress, which affected the development of PM2.5-induced PF. [Display omitted] • Nrf2 defends against oxidative stress in PM2.5-exposed mouse-lung and 16HBE cells. • METTL3-mediated m6A-modified Nrf2 mRNA promotes Nrf2 translation after PM2.5 exposure. • The exact Nrf2-mRNA m6A sites that are involved in PM2.5-induced PF were identified. • YTHDF1/IGF2BP1 promoted Nrf2 translation by binding to m6A residues of Nrf2-mRNA. [ABSTRACT FROM AUTHOR]

The article discusses a study on 305 pulmonary arterial hypertension (PAH) patients in Shanghai Pulmonary Hospital that analyzed the influence of bone morphogenetic protein receptor type II (BMPR2) mutations on phenotype and whether this influence is associated with sex. The study found that after adjustment for age and sex, BMPR2 mutations were associated with a higher risk of mortality, and that the influence of BMPR2 mutations was more obvious in male PAH patients.

Abstract: Fibrodysplasia ossificans progressiva (FOP) is a rare autosomal dominant disorder characterized by progressive heterotopic bone formation in skeletal muscle tissue. Patients with FOP show malformed digits, osteochondroma, and other skeletal abnormalities due to abnormal patterning during development. Heterozygous mutations in the Activin A receptor type I (ACVR1) gene, which encodes the bone morphogenetic protein (BMP) type I receptor ALK2, have been identified in not only typical FOP patients but also patients with unusually mild or severe clinical features. The serine/threonine kinase activity of ALK2 may be constitutively activated by mutations in the GS domain or the kinase domain. Based on these findings, selective small chemical inhibitors and allele-specific RNAi approaches for mutant ALK2 have been developed for preventing heterotopic bone formation in FOP. Other novel treatments have also been reported to block heterotopic bone formation in patients with FOP. These findings open the door to the next step in FOP treatment and related research. [Copyright &y& Elsevier]

PURPOSE: To Investigate the mechanism of bone morphogenetic protein receptor (BMP-R) on inducing the osteogenesis of rat bone marrow stromal cells. METHODS: BMSCs were harvested from rats and induced in control medium (CM), osteogenic medium (OM), BMP-2, and OM with BMP-2. Alkaline phosphatase (ALP) activity assay and Von Kossa staining were used to assess the differentiation. BMP receptor (BMP-R) was also assessed using RT-PCR.SPSS 16.0 software package was used for data analysis with one-way ANOVA. RESULTS: BMSCs underwent osteogenic differentiation in OM group, showing high level of ALP activity and matrix mineralization. BMP-2 increased osteogenesis of BMSCs induced by OM. Significant BMP-R I A, BMP-R II expression could be detected in BMSCs in OM after 3 days of stimulation, while BMP receptors were later detected in BMSCs in BMP-2. CONCLUSIONS: BMP-2 can increase osteogenesis of BMSCs induced by OM. BMP receptors have a strong effect on the osteogenic differentiation of BMSCs. Supported by Natural Science Foundation of Guangdong Province (9151008901000035) and Project of Science and Technology plan of Guangdong Province (200813080701002). [ABSTRACT FROM AUTHOR]

Lipoprotein (a) [Lp(a)] is a causal risk factor for cardiovascular diseases, while its role in vascular calcification has not been well-established. Here, we investigated an association of Lp(a) with vascular calcification using population-based and in vitro study designs. A total of 2806 patients who received coronary computed tomography were enrolled to assess the correlation of Lp(a) with the severity of coronary artery calcification (CAC). Human aortic smooth muscle cells (HASMCs) were used to explore mechanisms of Lp(a)-induced vascular calcification. In the population study, Lp(a) was independently correlated with the presence and severity of CAC (all p < 0.05). In vitro study showed that cell calcific depositions and alkaline phosphatase (ALP) activity were increased and the expression of pro-calcific proteins, including bone morphogenetic protein-2 (BMP2) and osteopontin (OPN), were up-regulated by Lp(a) stimulation. Interestingly, Lp(a) activated Notch1 signaling, resulting in cell calcification, which was inhibited by the Notch1 signaling inhibitor, DAPT. Lp(a)-induced Notch1 activation up-regulated BMP2-Smad1/5/9 pathway. In contrast, Noggin, an inhibitor of BMP2-Smad1/5/9 pathway, significantly blocked Lp(a)-induced HASMC calcification. Notch1 activation also induced translocation of nuclear factor-κB (NF-κB) accompanied by OPN overexpression and elevated inflammatory cytokines production, while NF-κB silencing alleviated Lp(a)-induced vascular calcification. Elevated Lp(a) concentrations are independently associated with the presence and severity of CAC and the impact of Lp(a) on vascular calcification is involved in the activation of Notch1-NF-κB and Notch1-BMP2-Smad1/5/9 pathways, thus implicating Lp(a) as a potential novel therapeutic target for vascular calcification. • Lp(a) levels are independently associated with the presence and severity of CAC. • Lp(a) activates Notch1 signal to result in smooth muscle cell calcification. • The Lp(a)-induced Notch1 activation up-regulates BMP2-Smad1/5/9 signaling pathway. • Lp(a) mediates Notch1-NF-κB pathway with inflammatory response in cell calcification. [ABSTRACT FROM AUTHOR]

Bones are an important source of DNA for identification in forensic medicine, especially when the remains are skeletonized, which is the case when dealing with victims of the Second World War. Often the amount of bone available for sampling is limited, and therefore it is crucial to sample the bone segment with the highest adequate DNA quantity for identification. Studies performed on all representative skeletal element types of the human body showed that the amount of DNA obtained from different skeletal elements of different body regions varies greatly. When bones from torso were analyzed, thoracic vertebrae outperformed other vertebrae (cervical and lumbar) and, alongside the first ribs, were among the most appropriate bone elements for identification purposes. It was also shown that the quantity of DNA varies significantly within a single bone type. This study focused on exploring intra-bone DNA variability between five parts of 12th thoracic vertebrae (laminae + spinous process, pedicles + transverse processes, and corpus right, left, and middle). The research was based on the theory that the distribution of body weight and consequently bone remodeling, as well as the ratio between cancellous and cortical bone, contribute to different quantities of DNA in different parts of vertebra sampled. The vertebrae were cleaned and cut into five parts, and each part was completely ground to obtain homogenous bone powder. Half a gram of powder from each part was decalcified using a full demineralization extraction method. The DNA was purified in a Biorobot EZ1 machine (Qiagen). DNA quantity and quality were determined using the PowerQuant System (Promega) and autosomal STR typing success using the GlobalFiler Amplification Kit (Applied Biosystems). Thirty-five 12th thoracic vertebrae were sampled from a single Second World War mass grave. The best results with the highest DNA quantity were found in laminae and the spinous process, and among them all vertebrae analyzed yielded full STR profiles except three, where only a few dropouts occurred. The second-ranked bone part was the pedicles and transverse processes. The comparison of DNA degradation in the vertebral segments analyzed does not show statistically significant differences. Considering our research, when only the torso is available for identification, the 12th thoracic vertebra should be collected and the vertebral arch should be sampled for genetic analyses. • Higher DNA yield is present in areas with the highest strain or weight bearing due to a higher degree of bone remodeling. • More DNA was obtained from the vertebral arch than the vertebral body. • DNA quality (Auto/Deg ratio) did not vary across the 12th vertebra. • In all lamina + spinous process segments, full or almost full genetic profiles were obtained. • When only the torso is present, the vertebral arch of the 12th vertebra should be considered for DNA identification. [ABSTRACT FROM AUTHOR]

DNA yield and STR typing success differ among skeletal element types and within individual bones. Consequently, it is necessary to identify the skeletal elements and their intra-skeletal parts that will most likely yield utilizable and informative endogenous DNA for human identification of skeletal remains. The petrous portion of the temporal bone has been shown to be the most suitable skeletal part for sampling archaeological skeletons, and it has also been used successfully in some forensic cases. When all representative bone types were analyzed for three complete Second World War skeletons, metatarsals and metacarpals yielded more DNA than petrous bones (which generated full profiles even if the DNA yield was lower) and, among almost 200 Second World War metatarsals and metacarpals analyzed, metacarpals III were found to be the highest-yielding bones. To further improve the sampling strategy in DNA analysis of aged skeletal remains, a comparison between 26 petrous bones and 30 metacarpals III from Second World War skeletons was made considering intra-bone DNA yield variability. In metacarpals III only epiphyses were sampled, and in petrous bones only the dense part within the otic capsule was used. To exclude the influence of taphonomic issues as much as possible, petrous bones and metacarpals III from a single Second World War mass grave were examined. The difference between petrous bones and metacarpals III was explored by measuring nuclear DNA yield and success of STR typing. After cleaning the samples, full demineralization extraction was used to decalcify 0.5 g of powdered bone. PowerQuant (Promega) was used to determine DNA content and DNA degradation rates, and STR typing was performed using the PowerPlex ESI 17 Fast System (Promega). Metacarpals III produced the same DNA yields and STR typing success as petrous bones with no intra-individual difference observed in concentration of DNA, degradation rate, percentage of successfully amplified alleles, and intensity of electrophoretic signals. Sampling and processing of metacarpal III epiphyses is consequently recommended for genetic identification of highly degraded skeletal remains in routine forensic casework and in buried non-commingled aged skeletal remains whenever metacarpals III are preserved. Metacarpals III are easy to sample and less prone to contamination with modern DNA because no saw is needed for sampling in comparison to the petrous portion of the temporal bone. The data obtained in this study further improve the sampling strategy for genetic identification of Second World War skeletal remains in Slovenia. • Petrous bones are the most suitable skeletal part for sampling ancient skeletons. • Metacarpals III yielded the most DNA in Slovenian Second World War skeletons. • Second World War petrous bones were compared with metacarpals III, and intra-bone DNA variability was considered in sampling. • Metacarpals III produced the same DNA yields and STR typing success as petrous bones. • Whenever possible, metacarpals should be collected and epiphyses used for DNA identification. [ABSTRACT FROM AUTHOR]

The antioxidant activity in vivo and in vitro of peptides with <1 kDa molecular weight were measured after purification of the enzymatic hydrolysates (FlavourzymeTM and Papain PSM500 together) of porcine bone protein extracts (EH-PBPE). Amino acid sequences of peptides responsible for its characteristic umami taste were identified as Ser-Tyr, Pro-Asn, Gly-Ser, Lys-Pro, and Ala-Pro-His-Arg. The umami compounds were purified using ultrafiltration, gel filtration chromatography, preparative high-performance liquid chromatography, and liquid chromatography-tandem mass spectrometry (LC-MS); and sensory evaluations were done. The in vitro antioxidant activity (as DPPH radical-scavenging activity) of the peptide was 62.9% and umami score (estimated using sensory evaluation) was 8.9 out of 10. Subsequent in vivo studies on antioxidant activities (superoxide dismutase activity and malondialdehyde, glutathione, and protein carbonyl content) of the <1 kDa fraction showed a significant difference from those of the control group, in a dose-dependent manner. In total, 5 peptides were found in the <1 kDa fraction of EH-PBPE using LC-MS. These results suggested that the <1 kDa peptide fraction purified from EH-PBPE might be used as a natural antioxidant to offset oxidative stress. • Antioxidant activity and sensory evaluation of <1 kDa PBPE peptides were carried out. • In vivo experiments showed changes in GSH, MDA, and protein carbonyl content of these peptide. • Amino acid sequence of the peptide fraction (<1 kDa) of EH-PBPE was obtained. [ABSTRACT FROM AUTHOR]

Tibial Dyschondroplasia (TD) is a prevailing skeletal disorder that mainly affects rapidly growing avian species. It results in reduced bone strength, lameness and an increase risk of fragility fractures. Total flavonoids of Rhizoma drynariae (TFRD) have been used as an effective treatment of different bone diseases in humans. The current in vitro study was conducted to explore the therapeutic effect of TFRD on thiram-induced cytotoxicity in avian growth plate cells via bone morphogenetic protein-2/runt related transcription factor-2 (BMP-2/Runx2) and Indian hedgehog/Parathyroid hormone-related peptide (IHH/PTHrP) expressions. Chondrocytes were isolated, cultured and refined from chicken's tibial growth plates in a special medium. Then chondrocytes were treated with sublethal thiram having less concentration (2.5 μg/mL) to induce cytotoxicity of chondrocyte, and then treated with providential doses (100 μg/mL) of TFRD. Thiram caused distorted morphology of chondrocytes, nuclei appeared disintegration or lysed along with decreased expressions of BMP-2/Runx2 and IHH/PTHrP. TFRD administration not only enhanced the viability of chondrocytes by itself, but also well restored the damage caused by thiram on growth plate chondrocytes by significantly up-regulating the expressions of BMP-2/Runx2 and IHH/PTHrP. Therefore, this study provides a novel insight into the further treatment of TD and other skeletal ailments and lays the foundation for prevention and treatment. • TD is a prevailing skeletal disorder affecting rapidly in poultry industry. • TD is associated with toxic effect of thiram due to contaminated environment. • TFRD have been used as an effective treatment of different bone diseases. • TFRD administration enhances the viability of growth plate chondrocytes. • TFRD significantly up-regulates the expressions of BMP-2/Runx2 & IHH/PTHrP. [ABSTRACT FROM AUTHOR]

Body deformity occurs both in wild and farmed fishes, which is one of the most challenging problems for aquaculture industry. In most cases, such body deformities are linked to skeletal deformities. Currently, very limited information is available on skeletal deformities of farmed fish species which may be caused by genetic factor. In this study, we performed muscle and vertebra transcriptome analyses in body deformity and normality of bighead carp Hypophthalmichthys nobilis (from one meiotic gynogenesis family) using RNA-Seq. A total of 43,923 and 44,416 unigenes were predicted in muscles and vertebrae, respectively. Based on these data, we further explored the gene expression profiles in gynogenetic normal and abnormal bighead carp. No differentially expressed gene (DEG) was found in transcriptome data of muscles. Totally, 20 key DEGs were identified in transcriptome data of vertebrae, such as low density lipoprotein-related protein 2 (lrp2) , bone morphogenetic protein 2B (bmp2b) and collagen alpha-1(IV) (col4a1). 12 potential pathways were also identified in vertebra transcriptome data, which were mainly involved in development, growth, cytoskeleton and energy metabolism, such as MAPK signaling pathway, regulation of actin cytoskeleton and TGF-beta signaling pathway. Results of this study will be informative for the understanding of genetic mechanisms for body shape formation and also provide potential candidate genes for selection program involved in body shape and skeletal development in H. nobilis. Unlabelled Image • Body deformity is a threat to seed production of aquaculture. • We performed muscle and vertebra transcriptome analyses for body deformity of bighead carp. • Totally, 20 key DEGs and 12 potential pathways that may be involved in skeletal development were identified. • These results will be informative for studying genetic mechanisms of skeleton formation and deformity in bighead carp. [ABSTRACT FROM AUTHOR]

• BMP2, in combination with FGF2, not only enhanced the proliferation of bone marrow-derived MSCs but also strengthened their osteogenic potential after short-term expansion in vitro. • Both BMP and FGF signals are necessary and sufficient for the long-term expansion of MSCs with high osteogenic potential. Mesenchymal stem cells (MSCs) are a potential source of osteoblasts for the treatment of osteoporosis, but how to better preserve the stemness of MSCs in vitro culture conditions is the main challenge for MSC transplantation. The use of fibroblast growth factor 2 (FGF2) supplement has been described and used extensively to increase the expansion of MSCs. Cumulative evidence indicates that bone morphogenetic protein 2 (BMP2; a member of the TGF-β superfamily) is a secreted protein that promotes bone formation, which can regulate cell growth, differentiation, and development. Here we found that BMP2, in combination with FGF2, not only enhanced the proliferation of Macaca bone marrow-derived MSCs but also strengthened their osteogenic potential after short-term expansion in vitro. During long-term expansion, these cells still retained their osteogenic potential as well as other functional characteristics of pluripotent MSCs, which are gradually lost in the absence of BMP2. In addition, the BMP antagonist Noggin did not affect MSC expansion and the osteogenic potential. This study demonstrates that the regulation of BMP signaling can maintain the effectiveness of MSCs during expansion, which promotes the clinical application of MSCs in bone repair. [ABSTRACT FROM AUTHOR]

• We cultured canine PDLSCs and DPSCs successfully, which had self-renewal and multilineage differentiation ability. • Canine PDLSCs and DPSCs showed weaker proliferation and migration capacity than human cells. • After osteogenic induction, ALP was upregulated in human stem cells and canine PDLSCs instead of canine DPSCs. • Osteogenic genes, such as RUNX2 and BMP2, were upregulated later in induced canine cells than human cells. The effectiveness of stem cell-based periodontal tissue engineering need to be assessed by preclinical animal studies. Dog models are widely used animal models; however, there are not sufficient data on characterization of canine dental mesenchymal stem cells. Therefore, we aimed to compare the characteristics among canine and human periodontal ligament stem cells and canine and human dental pulp stem cells. Canine periodontal ligament stem cells and dental pulp stem cells showed significantly weaker clonogenic capability, and proliferation and migration capacity, and they displayed lower positive rates for CD90, CD73, CD105, and STRO-1. All of these canine and human cells showed multilineage differentiation potential. After osteogenic induction, the expression of alkaline phosphatase was obviously upregulated in human dental mesenchymal stem cells, but it was not upregulated in canine dental pulp stem cells. Other osteogenic genes, such as runt-related transcription factor 2 and bone morphogenetic protein 2, were upregulated in all induced canine and human cells, but their upregulation occurred later in canine cells. These results confirmed the stem cell properties of canine mesenchymal stem cells, but also suggested that more attention should be paid to the choice of appropriate research approaches, osteogenic gene markers, and time points for the utilization of canine dental mesenchymal stem cells due to their distinct characteristics. [ABSTRACT FROM AUTHOR]

Benzo[a]pyrene(BaP), a polycyclic aromatic hydrocarbons (PAH) of environmental pollutants, is one of the main ingredients in cigarettes and an agonist of the aryl hydrocarbon receptor (AhR). Mesenchymal stem cells (MSCs) including C3H10T1/2 and MEF cells, adult multipotent stem cells, can be differentiated toward osteoblasts during the induction of osteogenic induction factor-bone morphogenetic protein 2(BMP2). Accumulating evidence suggests that BaP decreases bone development in mammals, but the further mechanisms of BaP on BMP2-induced bone formation involved are unknown. Here, we researched the role of BaP on BMP2-induced osteoblast differentiation and bone formation. We showed that BaP significantly suppressed early and late osteogenic differentiation, and downregulated the runt-related transcription factor 2(Runx2), osteocalcin(OCN) and osteopontin (OPN) during the induction of BMP2 in MSCs. Consistent with in vitro results, administration of BaP inhibited BMP2-induced subcutaneous ectopic osteogenesis in vivo. Interestingly, blocking AhR reversed the inhibition of BaP on BMP2-induced osteogenic differentiation, which suggested that AhR played an important role in this process. Moreover, BaP significantly decreased BMP2-induced Smad1/5/8 phosphorylation. Furthermore, BaP significantly reduced bone morphogenetic protein receptor 2(BMPRII) expression and excessively activated Hey1. Thus, our data demonstrate the role of BaP in BMP2-induced bone formation and suggest that impaired BMP/Smad pathways through AhR regulating BMPRII and Hey1 may be an underlying mechanism for BaP inhibiting BMP2-induced osteogenic differentiation. BMP2 binding to BMPR-II to phosphorylate Smads(including Smad1/5/8) and then shuttles in the nucleus to regulate the expression of osteogenic gene. BaP crosses cell membrane and binds to AhR, inhibiting the expression of BMPR-II, thereby inhibiting the activation of Smad1/5/8, and finally inhibiting the expression of Runx2, OCN, and OPN. In addition, BaP excessively activate Hey1. This may eliminate the activation of the BMP signaling pathway through negative feedback, then negatively regulate BMP2-induced osteogenesis. Image 1 • BaP inhibits BMP2/Smad signaling pathway via AhR in MSCs. • BaP inhibits BMP2-induced osteogenic differentiation of MSCs by regulating BMPRII and Hey1. • Our research provides a theoretical basis for improving the bone formation of BaP exposed crowd. [ABSTRACT FROM AUTHOR]

Functional articular cartilage regeneration remains challenging, and it is essential to restore focal osteochondral defects and prevent secondary osteoarthritis. Combining autologous stem cells with therapeutic medical device, we developed a bi-compartmented implant that could promote both articular cartilage and subchondral bone regeneration. The first compartment based on therapeutic collagen associated with bone morphogenetic protein 2, provides structural support and promotes subchondral bone regeneration. The second compartment contains bone marrow-derived mesenchymal stem cell spheroids to support the regeneration of the articular cartilage. Six-month post-implantation, the regenerated articular cartilage surface was 3 times larger than that of untreated animals, and the regeneration of the osteochondral tissue occurred during the formation of hyaline-like cartilage. Our results demonstrate the positive impact of this combined advanced therapy medicinal product, meeting the needs of promising osteochondral regeneration in critical size articular defects in a large animal model combining not only therapeutic implant but also stem cells. In this manuscript, we report the preclinical evaluation, in a big animal model, of a hybrid implant, composed of a living compartment based on stem cell spheroids and an underlying functionalized matrix with sustained release of therapeutics, as a candidate for treatments of articular cartilage lesions. Our technology approach fulfills the goals of regenerating more robust subchondral bone, in a shorter time frame. This would help to improve the therapeutic efficacy of the current, less sophisticated implants in the field of osteochondral regenerative surgery. Unlabelled Image [ABSTRACT FROM AUTHOR]

• TLR4 and TLR7 are expressed in the peripheral nervous system during embryonic development. • TLR4 and TLR7 expression show a different spatiotemporal distribution. • TLR4 is present in neurons of DRG and PVG by E14 and only at E18 in ENS while TLR7 starts to be detectable already at E12. Toll-Like Receptors (TLRs) play a critical role in the innate and adaptive immune system. They are the mammalian orthologs of Drosophila melanogaster protein Toll, which has been proved to have an early morphogenetic role in invertebrate embryogenesis that in the adult switches to an immune function. The aim of this study was to evaluate the expression of TLR4 and TLR7 during dorsal root ganglia (DRG), paravertebral ganglia (PVG), and enteric nervous system (ENS) murine development. Mouse embryos from different stages (i.e. E12 to E18) were processed for immunolocalization analysis on formalin-fixed paraffin-embedded sections, and isolated intestine were processed for whole-mount preparations. We observed a differentially regulated expression of TLR4 and TLR7 during embryogenesis and an overall increased expression of both receptors during development. While TLR4 was detectable in neurons of DRG and PVG starting from E14 and only from E18 in the ENS, TLR7 was already expressed in scattered neurons of all the investigated regions at E12. TLR4 and TRL7 expression temporal patterns suggest a morphogenetic role for these receptors in the development of neural crest derivatives in mammals. [ABSTRACT FROM AUTHOR]

Screening of bone morphogenetic protein receptor type- 1 gene (BMPR1B gene) also known as Fec B gene polymorphism was done for detecting the genetic basis of prolificacy in crossbred does of Bihar. The study revealed two allelic variants (A=0.48 and G=0.52) and three genotypes (AA=0.16, AG=0.62 and GG=0.22).The findings of thestudy indicate sign of introgression of mutant allele (G) into Jamunapari, Barbari and Sirohi goats at village level from Black Bengal breed. The variation in litter size among the genotypes revealed the benefits of introgression of Fec B mutation in the crosses. The litter size showed variation among the three genotypes. Percentage singles, twins and triplets were 69, 29 and 2 in AA genotype; 42, 52 and 6 in AG genotype and 14, 66 and 20 in GG genotype, respectively. [ABSTRACT FROM AUTHOR]

In Australia, some trees of the mangrove, Avicennia marina, growing in a chronic oil polluted site, produce chlorophyll deficient (albino) propagules. We tested the hypothesis that albinism was due to an oil-induced mutant allele that controls photosynthesis. We determined whether there are genetic differences between normal and chlorophyll deficient propagules. Four gene regions (nuclear 18S–26S cistron; chloroplast - trn H -psb A, rsp 16 and mat K) were sequenced and analysed for normal and albino propagules. Mutations occurred in both nuclear (ITS) and coding chloroplast (mat K) genes of albino propagules. There were 10 mutational differences between normal and albino propagules in the mat K samples. Analysis of molecular variation (AMOVA) of the mat K dataset indicated highly significant genetic differentiation between normal and albino propagules. Our study suggests for the first time that PAHs from a chronic oil polluted site resulted in mutations in both nuclear and chloroplast genes, resulting in the production of albino propagules. [ABSTRACT FROM AUTHOR]

Abstract: Juvenile polyposis of infancy is the most severe and life-threatening form of juvenile polyposis. This disease typically presents in the first two years of life with gastrointestinal bleeding, diarrhea, inanition, and exudative enteropathy. In very few reports concerning this entity, a large deletion in the long arm of chromosome 10 (10q23), encompassing the PTEN and BMPR1A genes, was found. The authors report a case of delayed diagnosis of juvenile polyposis of infancy at 6years of age. A 3.34Mb long de novo deletion was identified at 10q23.1q23.31, encompassing the PTEN and BMPR1A genes. The disease course was severe with diarrhea, abdominal pain, inanition, refractory anemia, rectal bleeding, hypoalbuminemia, and exudative enteropathy. A sub-total colectomy, combined with intraoperative endoscopic removal of ileal and rectal stump polyps, was required for palliative disease control. [Copyright &y& Elsevier]