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An accurate and non-invasive histologic assessment of renal masses remains elusive, as approximately 15% of resected renal masses prove benign upon resection. Oncocytomas and hybrid oncocytoma/chromophobe tumors (HOCTs) are the most commonly-resected oncologically low-risk masses and are not reliably distinguished from malignant lesions with standard cross-sectional imaging. Histologically, these tumors are notable for densely-packed mitochondria. Technetium-99m-sestamibi single-photon emission CT/ x-ray CT is a nuclear imaging modality that utilizes a mitochondria-targeting tracer. Early series using 99mTc-sestamibi SPECT/CT showed promise in differentiating renal cell carcinoma (RCC) from oncocytomas and HOCTs, but recently published data from our institution found that oncocytomas still accounted for 20% of lesions with low 99mTc-sestamibi tracer uptake, as qualitatively interpreted by a senior nuclear radiologist. Here, we asses performance of 99mTc-sestamibi scans utilizing quantitative assessment thresholds, hypothesizing that this may improve test characteristics over qualitative assessments. Patients undergoing 99mTc-sestamibi SPECT/CT for evaluation of renal masses between February 2020 and December 2021 were included in our analysis. A mass was labeled as "hot" by the radiologist if its 99mTc-sestamibi uptake was qualitatively equivalent or higher than that of the ipsilateral renal parenchyma; otherwise it was labeled as "cold". Using nuclear medicine image processing software, TBRs were calculated by comparing signal counts of the masses to those of the normal ipsilateral renal parenchyma using manually generated regions of interest. Quantitative "hot"/"cold" determinations for each mass were then re-evaluated using previously published TBR cutoffs of 0.46 and 0.6. Masses for which TBRs could not be calculated were assumed to have unchanged classifications on quantitative analysis. Qualitative and quantitative 99mTc-sestamibi SPECT/CT findings were then correlated to histology for masses that underwent pathologic confirmation via biopsy or surgical excision. 78 patients underwent 99mTc-sestamibi SPECT/CT for 98 renal masses. 52 masses had diagnostic pathology available from biopsy or surgical excision. Of these, 7 were "hot" (1 RCC, 6 oncocytomas) and 45 were "cold" (34 RCC, 2 non-RCC malignancies, 9 oncocytomas). The negative predictive value of qualitatively-interpreted "cold" 99mTc-sestamibi scans for ruling out oncocytoma was 80%. 1/52 (1.9%) malignant masses were interpreted as "hot" and;likely benign. When a TBR cutoff of 0.46 was applied, 17/45 "cold" masses were reclassified as "hot" (13 RCC, 1 non-RCC malignancy, 6 oncocytomas). For a TBR cutoff of 0.6, 6/45 "cold" masses were reclassified as "hot" (1 RCC, 1 non-RCC malignancy, 4 oncocytomas) while 1 "hot" RCC was reclassified as "cold" (Figure 1).; TBR cutoffs of 0.46 and 0.6 improved the NPV;of 99mTc-sestamibi SPECT/CT to 89% and 88% but increased the rate of "hot" malignant tumors to 23.1% and 3.8%, respectively. When assessing histology of renal masses, applying previously-published quantitative TBR cutoffs to 99mTc-sestamibi SPECT/CT provides <10% improvement in the negative predictive value for ruling out oncocytomas compared to previously published qualitative interpretations of these scans. Conversely, the use of these thresholds in place of qualitative assessments tends to categorize more RCCs as "hot," thereby erroneously mimicking oncocytomas. Further work is needed to determine whether 99mTc-sestamibi SPECT/CT has a role in routine clinical practice. [ABSTRACT FROM AUTHOR]

Sensitivity analysis is popular in dealing with missing data problems particularly for non-ignorable missingness, where full-likelihood method cannot be adopted. It analyses how sensitively the conclusions (output) may depend on assumptions or parameters (input) about missing data, i.e. missing data mechanism. We call models with the problem of uncertainty sensitivity models. To make conventional sensitivity analysis more useful in practice we need to define some simple and interpretable statistical quantities to assess the sensitivity models and make evidence based analysis. We propose a novel approach in this paper on attempting to investigate the possibility of each missing data mechanism model assumption, by comparing the simulated datasets from various MNAR models with the observed data non-parametrically, using the K-nearest-neighbour distances. Some asymptotic theory has also been provided. A key step of this method is to plug in a plausibility evaluation system towards each sensitivity parameter, to select plausible values and reject unlikely values, instead of considering all proposed values of sensitivity parameters as in the conventional sensitivity analysis method. The method is generic and has been applied successfully to several specific models in this paper including meta-analysis model with publication bias, analysis of incomplete longitudinal data and mean estimation with non-ignorable missing data. [ABSTRACT FROM AUTHOR]

In attempt to evaluate the effects of cyclodextrins (CDs) on enantioselectivity of chiral pesticides toxicity, this study investigated effects of three kinds of cyclodextrins including α-CD, β-CD and randomly methylated β-CD (RAMEB) on toxicity of four enantiomers of isomalathion including (1R, 3R)-isomalathion, (1S, 3S)-isomalathion, (1S, 3R)-isomalathion and (1R, 3S)-isomalathion. Generally, the addition of α-CD and RAMEB (1.5 g/L to 3.5 g/L concentration) could lead to reduction of isomalathion toxicity in most cases, while the presence of β-CD (0.3 g/L to 1.5 g/L concentration) enhanced the toxicity of isomalathion. It was speculated that higher electronic cloud density and lower water solubility of β-CD than α-CD and RAMEB might favor to combination between acetylcholinesterase (AChE) and isomalathion included by β-CD. With respect for α-CD and RAMEB, isomalathion included by them could be easily dissolved in water because of high water solubility of the two CDs. Therefore, α-CD and RAMEB can be used as remediation regent for the pollution of isomalathion, and β-CD can act as an additive in improving bioactivity of such pesticides. In addition, the presence of CDs can alter enantioselectivity of chiral pesticides. The differences on the extent of enantioselectivity variation of isomalathion induced by α-CD, RAMEB and β-CD might be ascribe to the different cavity, electron cloud density and solubility among the three CDs. In conclusion, the above results gave researchers a possibility to change enantioselectivity of chiral pesticides from undesirable outcomes to desirable ones. [ABSTRACT FROM AUTHOR]

Above-deck car-bomb attacks cause significant effects on engineering structures from those of explosive blasts in the free air. In this paper, a post-disaster investigation for an existing bridge under above-deck car-bomb blast loading is presented. The crack distribution and damage deformation of the blast loaded girders are systematically investigated, and the blast effect zone is proposed to describe the blast loading distributions on girders and to characterize the girder structural behavior. The girder failure is dominated by one of the three typical damage mechanisms: shear, flexural-shear, and flexural, depending on the blast effect zone where the girder is located. The study also indicates that the isolation effect of the vehicle can significantly affect the blast loading distribution on structures, even though the isolation effect is localized in a region with a small standoff distance. It is gradually attenuated by the increase of the standoff distance. Numerical analysis is conducted to verify the findings by simulating the truck isolation effect with a steel plate. [ABSTRACT FROM AUTHOR]

[Display omitted] • The uncertainty and reliability of hybrid energy system are investigated. • A multi-objective mathematical framework is formulated to enhance the reliability. • The Pareto frontier is obtained using global optimization solvers. • A reliability-gain index is presented to evaluate the trade-off solutions. • The reliability is improved by 44% while the cost is only increased by 5%. More and more attention has been paid to the industrial hybrid energy systems which comprise fossil energy and renewable energy for achieving carbon neutrality. Wherein, the uncertainty of renewable energy, fluctuation of energy requirement from process units, and failure rate of energy conversion devices have significant effects on the reliability of industrial hybrid energy systems and thus restrict the application and promotion. How to simultaneously address these issues for realizing the reliable design of industrial hybrid energy systems has been a matter of great concern and challenge. Hence, a systematic reliable design optimization method is presented in this study to tackle these challenges. In this study, the two-stage stochastic programming method is implemented to conduct the uncertain optimization in dealing with the uncertainty of renewable energy and fluctuation of energy requirement, while a mixed stand-by redundancy allocation problem is proposed for simultaneously enhancing the reliability and minimizing the number of stand-by devices. A multi-objective mixed integer non-linear programming framework using reliability and total annual cost as the dual objectives is then formulated and applied to a refinery site for the reliable design optimization of the industrial hybrid energy system. According to the solution results, compared to the minimum total annual cost scheme without stand-by redundancy, the maximum reliability of the industrial hybrid energy system can reach 0.9878 using the stand-by redundancy strategy. As a result, the reliability can be significantly enhanced by 44%, with only a 5% increment of the total annual cost. Moreover, a Pareto frontier under multi-objective optimization is constructed, and the optimization schemes at the Pareto frontier can realize the trade-off between reliability and the total annual cost of the industrial hybrid energy system. Subsequently, a reliability-gain index is proposed to further evaluate the trade-off schemes and thus guide the choice of appropriate design schemes. [ABSTRACT FROM AUTHOR]

Neuroendocrine tumors (NETs) occur in various regions of the body and present with complex clinical and biochemical phenotypes. The molecular underpinnings that give rise to such varied manifestations have not been completely deciphered. The management of neuroendocrine tumors (NETs) involves surgery, locoregional therapy, and/or systemic therapy. Several forms of systemic therapy, including platinum-based chemotherapy, temozolomide/capecitabine, tyrosine kinase inhibitors, mTOR inhibitors, and peptide receptor radionuclide therapy have been extensively studied and implemented in the treatment of NETs. However, the potential of immune checkpoint inhibitor (ICI) therapy as an option in the management of NETs has only recently garnered attention. Till date, it is not clear whether ICI therapy holds any distinctive advantage in terms of efficacy or safety when compared to other available systemic therapies for NETs. Identifying the characteristics of NETs that would make them (better) respond to ICIs has been challenging. This review provides a summary of the current evidence on the value of ICI therapy in the management of ICIs and discusses the potential areas for future research. [ABSTRACT FROM AUTHOR]

Abstract: Although recent studies have established that osteocytes function as secretory cells that regulate phosphate metabolism, the biomolecular mechanism(s) underlying these effects remain incompletely defined. However, investigations focusing on the pathogenesis of X-linked hypophosphatemia (XLH), autosomal dominant hypophosphatemic rickets (ADHR), and autosomal recessive hypophosphatemic rickets (ARHR), heritable disorders characterized by abnormal renal phosphate wasting and bone mineralization, have clearly implicated FGF23 as a central factor in osteocytes underlying renal phosphate wasting, documented new molecular pathways regulating FGF23 production, and revealed complementary abnormalities in osteocytes that regulate bone mineralization. The seminal observations leading to these discoveries were the following: 1) mutations in FGF23 cause ADHR by limiting cleavage of the bioactive intact molecule, at a subtilisin-like protein convertase (SPC) site, resulting in increased circulating FGF23 levels and hypophosphatemia; 2) mutations in DMP1 cause ARHR, not only by increasing serum FGF23, albeit by enhanced production and not limited cleavage, but also by limiting production of the active DMP1 component, the C-terminal fragment, resulting in dysregulated production of DKK1 and β-catenin, which contributes to impaired bone mineralization; and 3) mutations in PHEX cause XLH both by altering FGF23 proteolysis and production and causing dysregulated production of DKK1 and β-catenin, similar to abnormalities in ADHR and ARHR, but secondary to different central pathophysiological events. These discoveries indicate that ADHR, XLH, and ARHR represent three related heritable hypophosphatemic diseases that arise from mutations in, or dysregulation of, a single common gene product, FGF23 and, in ARHR and XLH, complimentary DMP1 and PHEX directed events that contribute to abnormal bone mineralization. This article is part of a Special Issue entitled "The Osteocyte". [Copyright &y& Elsevier]

Differences in siphonophore communities between summer and winter in the northern South China Sea and the influence of environmental conditions on their variations were studied. Our results suggested that local coastal upwelling and surface ocean currents driven by the southwest monsoon enhanced the species number and abundance of siphonophores in summer, while the northeast monsoon forced the cold coastal current into the study area, resulting in low species richness and abundance in winter. [ABSTRACT FROM AUTHOR]

We consider a Gaussian process regression (GPR) approach to analysing a single-index model (SIM) from the Bayesian perspective. Specifically, the unknown link function is assumed to be a Gaussian process a priori and a prior on the index vector is considered based on a simple uniform distribution on the unit sphere. The posterior distributions for the unknown parameters are derived, and the posterior inference of the proposed approach is performed via Markov chain Monte Carlo methods based on them. Particularly, in estimating the hyperparameters, different numerical schemes are implemented: fully Bayesian methods and empirical Bayes methods. Numerical illustration of the proposed approach is also made using simulation data as well as well-known real data. The proposed approach broadens the scope of the applicability of the SIM as well as the GPR. In addition, we discuss the theoretical aspect of the proposed method in terms of posterior consistency. [ABSTRACT FROM AUTHOR]

Osteoarthritis (OA) is considered a multifactorial disease whose development and progression may include several structural abnormalities aside from cartilage destruction. Bone marrow lesions (BMLs) have been reported to be associated with OA pathology, and several studies have advocated its close connection to the severity of joint structural alterations and pain, the main OA clinical manifestation. Hence, BMLs may not only affect subchondral bone and its neuronal and vascular structures but also negatively influence the adjacent tissues. Here, we analyze the pathophysiology and natural history of OA-associated BMLs and their potential relevance to the radiographic progression and severity of the disease. The notion that BMLs may be a precursor to additional articular abnormalities, can be a potential risk factor for development of OA, and may serve as an additional diagnostic tool and a therapeutic target are further discussed. [ABSTRACT FROM AUTHOR]

Abstract: This study examines species composition and abundance of appendicularians in the northwestern coastal waters of South China Sea based on in situ data obtained by 169μm planktonic nets during summer and winter, and discusses the influence of environmental factors on their distribution and assemblages. A total of 19 appendicularian species (including one unidentified species) was collected. Species richness is higher in summer than in winter, and it increases from inshore to offshore waters. Average abundance of appendicularians is 103.1 (±198.1) and 54.6 (±111.1)ind.m−3 in summer and winter, respectively. The abundance distribution of appendicularians is generally high around inshore waters during summer and winter, and with a higher abundance assemblage along the eastern coast from Leizhou Peninsula to Hainan Island, especially at the eastern mouth of Qiongzhou Strait in summer. Oikopleurids accounted for 97.5% and 99.6% of total appendicularians abundance in summer and winter, and the region of high values was the same to that of total appendicularians abundance. Oikopleura longicauda, Oikopleura rufescens, and Oikopleura fusiformis are dominant species during summer and winter. Results showed that spatial distribution of species richness of appendicularians is considered to be the result of physical factors such as temperature and salinity in summer and winter, while variations in abundance of appendicularians are affected by the combination of physical and biological factors. In particular, variations of appendicularians abundance are affected by food availability in summer. [ABSTRACT FROM AUTHOR]

The wear, delamination, and fatigue resistance of artificially aged gamma irradiation-sterilized conventional polyethylene (CPE) and gas-plasma-sterilized melt-annealed highly crosslinked polyethylene tibial inserts (HXPE) were compared. Six CPE and 12 HXPE (six irradiated at 58 kGy and six at 72 kGy) left knee inserts were wear tested for 5.5 million cycles (Mc) under loads and motions that mimic activities of daily living, such as walking, chair rise, stair ascent, and deep squatting. Another six HXPE (72 kGy) and six CPE inserts were also tested under conditions that could produce severe delamination for 8 Mc. Ten other knees (five 72 kGy HXPE and five CPE) were subjected to posterior edge loading fatigue testing for 5 Mc. The HXPE inserts had an average wear rate reduction of about 80% relative to their CPE counterparts during all activities. All of the CPE inserts delaminated and fractured during high cycle deep squat (152° flexion) motions, while all the HXPE remained intact. None of the HXPE inserts delaminated after 8 Mc, while all of the CPE inserts developed delamination damage within 1.5-5.8 Mc of delamination testing. All CPE inserts developed subsurface cracks and delamination within 2.8 Mc during posterior edge loading fatigue studies, while none of the HXPE inserts showed cracking or delamination after 5 Mc. These results show that aged HXPE has higher wear and fatigue resistance than aged CPE, and offers potential long-term advantages for young active patients with sustained activities of daily living. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:1120-1126, 2010 [ABSTRACT FROM AUTHOR]

Granulin epithelin precursor (GEP) has been implicated in development, tissue regeneration, tumorigenesis, and inflammation. Herein we report that GEP stimulates chondrocyte differentiation from mesenchymal stem cells in vitro and endochondral ossification ex vivo, and GEP-knockdown mice display skeleton defects. Similar to bone morphogenic protein (BMP) 2, application of the recombinant GEP accelerates rabbit cartilage repair in vivo. GEP is a key downstream molecule of BMP2, and it is required for BMP2-mediated chondrocyte differentiation. We also show that GEP activates chondrocyte differentiation through Erk1/2 signaling and that JunB transcription factor is one of key downstream molecules of GEP in chondrocyte differentiation. Collectively, these findings reveal a novel critical role of GEP growth factor in chondrocyte differentiation and the molecular events both in vivo and in vitro. [ABSTRACT FROM AUTHOR]

Abstract: Neisseria meningitidis is the etiologic agent of meningococcal meningitis. We compared 48-h biofilm formation by N. meningitidis serogroup B strains NMB, MC58, C311 and isogenic mutants defective in capsule formation on SV-40 transformed human bronchial epithelial (HBE) cells in a flow cell. We demonstrated that strains NMB and NMB siaA-D were defective in biofilm formation over glass, and there was a partial rescue of biofilm growth for strain NMB on collagen-coated coverslips at 48 h. We demonstrated all three serogroup B strains form biofilms of statistically equivalent average height on HBE cells as their isogenic capsular mutants. Strain NMB also formed a biofilm of statistically equivalent biomass as the NMB siaA-D mutant on HBE cells at 6 and 48 h. These biofilms are significantly larger than biofilms formed over glass or collagen. Verification that strain NMB expressed capsule in biofilms on HBE cells was demonstrated by staining with 2.2.B, a monoclonal antibody with specificity for the serogroup B capsule. ELISA analysis demonstrated that strains MC58 and C311 also produced capsules during biofilm growth. These findings suggest that encapsulated meningococci can form biofilms on epithelial cells suggesting that biofilm formation may play a role in nasopharyngeal colonization. [Copyright &y& Elsevier]

Abstract: Biofilms form in a variety of host sites following infection with many bacterial species. However, the study of biofilms in a host is hindered due to the lack of protocols for the proper experimental investigation of biofilms in vivo. Histophilus somni is an agent of respiratory and systemic diseases in bovines, and readily forms biofilms in vitro. In the present study the capability of H. somni to form biofilms in cardiopulmonary tissue following experimental respiratory infection in the bovine host was examined by light microscopy, transmission electron microscopy, immunoelectron microscopy of ultrathin cryosections, scanning electron microscopy of freeze-fractured samples, and fluorescent in situ hybridization. Biofilms were evident and most prominent in the myocardium, and were associated with a large amount of amorphous extracellular material. Furthermore, Pasteurella multocida was often cultured with H. somni from heart and lung samples. Transposon mutagenesis of H. somni strain 2336 resulted in the generation of mutants that expressed more or less biofilm than the parent strain. Six mutants deficient in biofilm formation had an insertion in the gene encoding for a homolog of filamentous haemagglutinin (FHA), predicted to be involved in attachment. Thus, this investigation demonstrated that H. somni is capable of forming a biofilm in its natural host, that such a biofilm may be capable of harboring other bovine respiratory disease pathogens, and that the genes responsible for biofilm formation can be identified by transposon mutagenesis. [Copyright &y& Elsevier]

Neisseria gonorrhoeae forms a biofilm inflowcells on glass coverslips as well as on primary cervical epithelial cells. Electron microscopic studies of cervical biopsy specimens from 10 patients with culture-proven N. gonorrhoeae infection revealed evidence of biofilm formation in 3 of the biopsy specimens. These biofilms showed gonococci in networks of bacterial membrane within the biofilm structure. This finding was also observed in biofilms formed over glass cover slips and after infection of primary cervical tissue in vitro. The importance of membranous networks in Neisseria biofilm formation was demonstrated with N. gonorrhoeae strain 1291-msbB, which shows a markedly decreased ability to bleb. This mutant formed significantly less biofilm over glass surfaces and cervical epithelial cells, and complementation showed reversion to wild-type biofilms. Gonoccal biofilms, as part of the cervical infection, may be involved in the mechanisms by which asymptomatic infections, persistence, and increased antibiotic resistance occur. [ABSTRACT FROM AUTHOR]

Abstract: The dynamic growth of algae is influenced by the inner physiological characteristics as well as the exterior stimulating factors, including light, temperature and nutrients. The suspended sediments release nutrients driven by hydrodynamic forces, imposing significant effects on algae growth. In this research, the mechanism of algae growth was studied, and an experiment on algae growth was conducted in an ecological flume using water and sediments collected from Taihu lake, an eutrophic lake in China, with three water current stages: still water stage, slow water current stage, and fast water current stage. Based on the experiment, a model was developed to quantitatively describe the relations between the release of nitrogen/phosphorous nutrients and the water flow velocity in nitrogen and phosphorus cycles. Detailed calibration and verification of the model parameters were carried out based on the results from experiments. The predicted values fit presented the results fitting well with the measured data, which indicates that the developed model can represent the dynamics of algae growth to a certain extent. This model has the potential for forecasting algal bloom in shallow, temperate lake systems. [Copyright &y& Elsevier]

The osteocyte, a terminally differentiated cell comprising 90%–95% of all bone cells, may have multiple functions, including acting as a mechanosensor in bone (re)modeling. Dentin matrix protein 1 (encoded by DMP1) is highly expressed in osteocytes and, when deleted in mice, results in a hypomineralized bone phenotype. We investigated the potential for this gene not only to direct skeletal mineralization but also to regulate phosphate (Pi) homeostasis. Both Dmp1-null mice and individuals with a newly identified disorder, autosomal recessive hypophosphatemic rickets, manifest rickets and osteomalacia with isolated renal phosphate-wasting associated with elevated fibroblast growth factor 23 (FGF23) levels and normocalciuria. Mutational analyses showed that autosomal recessive hypophosphatemic rickets family carried a mutation affecting the DMP1 start codon, and a second family carried a 7-bp deletion disrupting the highly conserved DMP1 C terminus. Mechanistic studies using Dmp1-null mice demonstrated that absence of DMP1 results in defective osteocyte maturation and increased FGF23 expression, leading to pathological changes in bone mineralization. Our findings suggest a bone-renal axis that is central to guiding proper mineral metabolism. [ABSTRACT FROM AUTHOR]

We have studied gonococcal infection in human endometrium organ culture and in human primary endometrial epithelial cells using various microscopic techniques including scanning electron microscopy, transmission electron microscopy, bright field light microscopy and laser scanning confocal microscopy. Here we describe the interactions betweenNeisseria gonorrhoeaeand human endometrial luminal epithelial cells at the ultrastructural levels.N. gonorrhoeaeattached to cilia but were not observed associated with the plasma membrane of ciliated epithelial cells or internalized into ciliated epithelial cells.N. gonorrhoeaecould be found in intracellular vacuoles in secretory epithelial cells.N. gonorrhoeaehave diverse interactions with endometrial epithelium. These include intimate association and colocalization with asialoglycoprotein receptor (ASGP-R) and CEACAM, lamellipodia and ruffle formation and colocalization with CR3, and microvillus engagement. These studies indicate thatN. gonorrhoeaeutilize multiple mechanisms to associate with endometrial epithelial cells and can associate with both ciliated and secretory cells. This diversity is consistent with a role of the endometrium as a transition zone between frequently asymptomatic cervical gonorrhoea and symptomatic pelvic inflammatory disease. [ABSTRACT FROM AUTHOR]

The non-covalent bond between aggrecan and hyaluronan is critical for maintaining the normal structure and function of the extracellular matrix in articular cartilage. The failure of this bond can cause the loss of aggrecan and destruction of the extracellular matrix of articular cartilage. In this study, the rupture force of the single bond between hyaluronan and hyaluronan binding protein – the complex of the hyaluronan binding region of aggrecan and link protein – was directly measured with a nanomechanical testing system as 40±11 pN. The results were compared to a theoretical prediction based on a smart version of the Monte Carlo simulation. [Copyright &y& Elsevier]

Wear evaluations of ultra-high molecular weight polyethylene (UHMWPE) materials for orthopedic bearings are typically conducted within a bovine serum solution environment, with the wear rate being characterized via mass loss measurements. The mass change of UHMWPE consists of not only mass loss due to wear but also mass increase due to fluid absorption. Indeed, with the highly crosslinked UHMWPE materials recently introduced for orthopedic implants, hip simulator evaluations often produce mass increases of the cup liners. This observation is due to the wear rate being lowered so significantly that it is outpaced by the rate of absorption of the surrounding fluid. Even when this fluid absorption is taken into consideration, the corrected rate of wear is still negative. However, a recent study by the present authors showed that, by lowering the load during the swing phase of the gait cycle, highly crosslinked UHMWPE liners exhibited a positive wear rate after correcting for the fluid absorption. In addition, the surface morphology of the liners indicated increased wear and there was also a concomitant, significantly increased fluid absorption. It was postulated that increased hydration might lead to increased wear of both conventional and highly crosslinked UHMWPE. In the present study, this hypothesis was tested by comparing the wear performance of UHMWPE on a serum-lubricated multidirectional pin-on-flat wear tester as a function of previously imposed fluid absorption. The UHMWPE pin samples were: non-irradiated; gamma irradiated in air (25–37 kGy); gamma irradiated in nitrogen (25–37 kGy); and electron-beam irradiated (100 kGy). It was found that hydration prior to wear testing did significantly increase wear of UHMWPE. Furthermore, highly crosslinked UHMWPE wore significantly less than conventional UHMWPE, regardless of the prior hydration condition. [Copyright &y& Elsevier]

Bone morphogenetic protein-2 (BMP-2) regulates growth plate chondrogenesis during development and postnatal bone growth, but the control mechanisms of BMP-2 expression in growth plate chondrocytes are unknown. Here we have used both in vitro and in vivo approaches to demonstrate that transcription factor, NF-κB, regulates BMP-2 gene expression in chondrocytes. Two putative NF-κB response elements were found in the -2712/+165 region of the BMP-2 gene. Cotransfection of mutant I-κBα expression plasmids with BMP-2 promoter-luciferase reporters into TMC-23 chondrocyte cell line suppressed BMP-2 transcription. Mutations in NF-κB response elements in the BMP-2 gene lead to decreases in BMP-2 promoter activity. Electrophoretic mobility shift assay using nuclear extracts from TMC-23 chondrocytic cells revealed that the NF-κB subunits p50 and p65 bound to the NF-κB response elements of the BMP-2 gene. Thus, NF-κB may positively regulate BMP-2 gene transcription. Consistent with these findings, expression of BMP-2 mRNA was significantly reduced in growth plate chondrocytes in NF-κB p50/p52 dKO mice, which associated with decreased numbers of 5-bromo-2′-deoxyuridine (BrdUrd)-positive cells in the proliferating zone of growth plate in these mice. Therefore, in postnatal growth plate chondrocytes, expression of BMP-2 is regulated by NF-κB, which may play an important role in chondrogenesis. [ABSTRACT FROM AUTHOR]

Examines the ultrastructural and biochemical localization of N-RAP at the interface between myofibrils and intercalated disks in the mouse heart. Specificity of N-RAP expression; Localization of N-RAP in adult hearts; Isolation of N-RAP in intercalated disks.

Background: Most peripheral afferent fibers involved in swallowing travel through the glossopharyngeal and vagus nerves and terminate in the nucleus of the tractus solitarius (NTS) and nodose ganglion (NG). Sensory neurons within the NTS and NG contain several neurotransmitters, including acetylcholine, histamine, serotonin and dopamine. The roles of these four neurotransmitters were investigated. Methods: The effects of atropine (muscarinic cholinergic receptor antagonist); pyrilamine maleate (PM, histamine H1 receptor antagonist); cimetidine (histamine H2 receptor antagonist); 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT, specific 5-HT1A receptor agonist); and selective dopamine D1 receptor antagonist (Sch-23390) on the number of swallows elicited by distilled water in anesthetized guinea pigs were investigated. Results: Atropine (0.2 mg/kg) inhibited swallowing by approximately 70%; PM (30 mg/kg) inhibited swallowing by approximately 60%; cimetidine (30 mg/kg) inhibited swallowing by approximately 52.9% and Sch-23390 (chronic treatment) inhibited swallowing by approximately 40%. In contrast, 8-OH-DPAT did not alter the number of swallows. Chronic pretreatment of Sch-23390 markedly decreased the substance P (SP) content in the pharyngeal mucosa and the esophagus. Conclusion: These findings indicate that acetylcholine, histamine and dopamine are involved in the regulation of the swallowing reflex, whereas it is unlikely that serotonin is involved. [ABSTRACT FROM AUTHOR]

The vertebrate musculoskeletal system is known to be formed by mesenchymal stem cells condensing into tissue elements, which then differentiate into cartilage, bone, tendon/ligament, and muscle cells. These lineage-committed cells mature into end-stage differentiated cells, like hypertrophic chondrocytes and osteocytes, which are expected to expire and to be replaced by newly differentiated cells arising from the same lineage pathway. However, there is emerging evidence of the role of cell transdifferentiation in bone development and disease. Although the concept of cell transdifferentiation is not new, a breakthrough in cell lineage tracing allowed scientists to trace cell fates in vivo. Using this powerful tool, new theories have been established: (1) hypertrophic chondrocytes can transdifferentiate into bone cells during endochondral bone formation, fracture repair, and some bone diseases, and (2) tendon cells, beyond their conventional role in joint movement, directly participate in normal bone and cartilage formation, and ectopic ossification. The goal of this review is to obtain a better understanding of the key roles of cell transdifferentiation in skeletal development and diseases. We will first review the transdifferentiation of chondrocytes to bone cells during endochondral bone formation. Specifically, we will include the history of the debate on the fate of chondrocytes during bone formation, the key findings obtained in recent years on the critical factors and molecules that regulate this cell fate change, and the role of chondrocyte transdifferentiation in skeletal trauma and diseases. In addition, we will also summarize the latest discoveries on the novel roles of tendon cells and adipocytes on skeletal formation and diseases. [ABSTRACT FROM AUTHOR]

Evaluates the role of endogenous and exogenous parathyroid hormone (PTH) on renal functions in perinatal chickens. Urine flow, glomerular filtration rate and relative clearance of electrolytes; Effects of calcium loading; Effects of ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid infusion; PTH administration; Effect of dibutyryl adenosine 3',5'-cyclic monophosphate.

Nebulin is a family of giant myofibrillar proteins with molecular masses ranging over 700-900 kDa, Using a human nebulin cDNA probe. we isolated three nebulin cDNA clones from a mouse skeletal muscle cDNA library These three clones, labeled 8c. 7a and 4b, carry inserts of 2.0. 3.0 and 3.5 kb, respectively. In Northern blots, each insert detected the same &ap;25 Kb message from skeletal muscle as the human nebulin probe. while detecting no messages from cardiac muscle. Sequence data in combination with reverse-transcriptase PCR indicates that Clones 7a and 8c overlap to form 4076 bp contiguous sequence Alignment with the published full-length human nebulin sequence indicates that clone 7a over 1596bp. Howover after the first 798-bp overlap the sequence of these two mouse nebulin clones diverge. suggesting that they derive from distinct transcripts encoding isoforms of mouse nebulin The mouse nebulin clones encode a series of &ap;245 residue super repeats. each of which can he subdivided into seven &ap;35-residue, weakly repeating modules centered around a conserved tyrosine residue, consistent with the human nebulin sequence. The mouse nebulin clones align along the central third of the full-length human sequence. corresponding to super repeats 8- 16 of the 22 super repeats found in human nebulin The translated sequence is greater than 90% identical to the human sequence. with the exception of a 200-amino-acid region at the C-terminus of done 4b which is less than 60% identical, In genomic Southern blots. a mouse nebulin probe detected a homologous sequence in a wide variety of vertebrate species under stringent conditions However, no significant hybridization was observed to genomic DNA from invertebrates and microorganisms. even under very low stringency The sequence and Southern-blot data suggest that the nebulin sequence is highly conserved among vertebrate species. [ABSTRACT FROM AUTHOR]

A modified version of the PGDx elioTM Plasma Resolve assay was validated as a laboratory-developed test (LDT) for clinical use in the Molecular Diagnostics Laboratory at Fox Chase Cancer Center. The test detects single nucleotide variants (SNVs) and small insertions and deletions (indels) in 33 target genes using fragmented genomic DNA extracted from plasma. The analytical performance of this assay was assessed with reference standard DNA and 29 samples from cancer patients and detected 66 SNVs and 23 indels. Using 50 ng of input DNA, the sensitivity was 95.5% to detect SNVs at 0.5% allele frequency, and the specificity was 92.3%. The sensitivity to detect indels at 1% allele frequency was 70.4%. A cutoff of 0.25% variant allele frequency (VAF) was set up for diagnostic reporting. An inter-laboratory study of concordance with an orthologous test resulted in a positive percent agreement (PPA) of 91.7%. [ABSTRACT FROM AUTHOR]

Osteocytes are the terminally differentiated bone cells resulted from bone formation. Although there are two distinct processes of bone formation, intramembranous and endochondral ossifications contributing to the formation of calvarial and long bones, it is not clear whether the distinct pathways determine the differences between calvaria and femoral cortical bone derived osteocytes. In the present study, we employed confocal structured illumination microscopy and mRNA‐sequencing analysis to characterize the morphologic and transcriptomic expression of osteocytes from murine calvaria and mid‐shaft femoral cortical bone. Structured illumination microscopy and geometric modelling showed round shaped and irregularly scattered calvarial osteocytes compared to spindle shaped and orderly arrayed cortical osteocytes. mRNA‐sequencing analysis indicated different transcriptomic profiles between calvarial and cortical osteocytes and provided evidence that mechanical response of osteocytes may contribute to geometrical differences. Furthermore, transcriptomic analysis showed that these two groups of osteocytes come from distinct pathways with 121 ossification‐related genes differentially expressed. Analysis of correlation between ossification and osteocyte geometries via a Venn diagram showed that several genes related to ossification, cytoskeleton organization and dendrite development were differentially expressed between calvarial and cortical osteocytes. Finally, we demonstrated that aging disrupted the organization of dendrites and cortical osteocytes but had no significant effects on calvarial osteocytes. Together, we conclude that calvarial and cortical osteocytes are different in various aspects, which is probably the consequence of their distinct pathways of ossification. [ABSTRACT FROM AUTHOR]

A major challenge in identifying candidates for nonoperative management of locally advanced rectal cancer is predicting pathologic complete response (pCR) following chemoradiation. We evaluated pre- and post-CRT PET-CT imaging to predict pCR and prognosis in this set of patients undergoing resection after neoadjuvant therapy. We retrospectively identified patients from 2002 to 2015 with locally advanced rectal cancer who underwent CRT, pre- and post-CRT PET-CT imaging, and resection. Univariate and multivariate analysis was performed and receiver operating characteristic (ROC) curves were generated to evaluate the association of PET-CT characteristics with pCR and survival. ROC curves were generated to define optimal cutoff points for predictive PET-CT characteristics. 125 patients were included. pCR rate was 28%, and follow-up was 48 mo. On multivariable analysis, patients who had a pCR had lower median post-CRT maximal standardized uptake value (SUV max) (3.2 versus 5.2, P = 0.009) and higher median %SUV decrease (72 versus 58%, P = 0.009). ROC curves were generated for %SUV max decrease (AUC = 0.70) and post-CRT SUV (AUC = 0.69). Post-CRT SUV max <4.3 and %SUV max decrease of >66% were equally predictive of pCR with a sensitivity of 65%, specificity of 72%, PPV of 44%, and NPV of 86%. Median 5-y overall and relapse-free survival were improved for patients with post-CRT SUV <4.3 (OS: 86 versus 66%, P = 0.01; RFS: 75 versus 52%, P = 0.01) or %SUV decrease of >66% (OS, 82 versus 66%, P = 0.05; RFS, 75 versus 54%, P = 0.01). PET/CT may be useful in identifying patients who did not achieve pCR, as well as overall survival in patients undergoing CRT for rectal cancer. Patients with a post-CRT SUV of >4.3 should be considered for operative management, as an estimated 86% of these patients will not have a pCR. [ABSTRACT FROM AUTHOR]

Background: The primary pathological alterations of Pendred syndrome are endolymphatic pH acidification and luminal enlargement of the inner ear. However, the molecular contributions of specific cell types remain poorly characterized. Therefore, we aimed to identify pH regulators in pendrin-expressing cells that may contribute to the homeostasis of endolymph pH and define the cellular pathogenic mechanisms that contribute to the dysregulation of cochlear endolymph pH in Slc26a4−/− mice. Methods: We used single-cell RNA sequencing to identify both Slc26a4-expressing cells and Kcnj10-expressing cells in wild-type (WT, Slc26a4+/+) and Slc26a4−/− mice. Bioinformatic analysis of expression data confirmed marker genes defining the different cell types of the stria vascularis. In addition, specific findings were confirmed at the protein level by immunofluorescence. Results: We found that spindle cells, which express pendrin, contain extrinsic cellular components, a factor that enables cell-to-cell communication. In addition, the gene expression profile informed the pH of the spindle cells. Compared to WT, the transcriptional profiles in Slc26a4−/− mice showed downregulation of extracellular exosome-related genes in spindle cells. Immunofluorescence studies in spindle cells of Slc26a4−/− mice validated the increased expression of the exosome-related protein, annexin A1, and the clathrin-mediated endocytosis-related protein, adaptor protein 2. Conclusion: Overall, cell isolation of stria vascularis from WT and Slc26a4−/− samples combined with cell type-specific transcriptomic analyses revealed pH-dependent alternations in spindle cells and intermediate cells, inspiring further studies into the dysfunctional role of stria vascularis cells in SLC26A4-related hearing loss. [ABSTRACT FROM AUTHOR]

Aim: To date, controversies still exist regarding the exact cellular origin and regulatory mechanisms of periodontium development, which hinders efforts to achieve ideal periodontal tissue regeneration. Axin2‐expressing cells in the periodontal ligament (PDL) have been shown to be a novel progenitor cell population that is essential for periodontal homeostasis. In the current study, we aimed to elucidate the regulatory role of bone morphogenetic protein receptor type 1A (BMPR1A)‐mediated BMP signalling in Axin2‐expressing cells during periodontium development. Materials and Methods: Two strains of Axin2 gene reporter mice, Axin2lacZ/+ and Axin2CreERT2/+; R26RtdTomato/+ mice, were used. We next generated Axin2CreERT2/+; R26RDTA/+; R26RtdTomato/+ mice to genetically ablate of Axin2‐lineage cells. Axin2CreERT2/+; Bmpr1afl/fl; R26RtdTomato/+ mice were established to conditionally knock out Bmpr1a in Axin2‐lineage cells. Multiple approaches, including micro‐computed tomography, calcein green, and alizarin red double‐labelling, scanning electron microscopy, and histological and immunostaining assays, were used to analyse periodontal phenotypes and molecular mechanisms. Results: X‐gal staining revealed that Axin2‐expressing cells in the PDL were mainly distributed along the alveolar bone and cementum surface. Cell lineage tracing and cell ablation assays further demonstrated the indispensable role of Axin2‐expressing cells in periodontium development. Next, we found that conditional knockout of Bmpr1a in Axin2‐lineage cells led to periodontal defects, which were characterized by alveolar bone loss, impaired cementogenesis, and abnormal Sharpey's fibres. Conclusions: Our findings suggest that Axin2‐expressing cells in the PDL are essential for periodontium development, which is regulated by BMP signalling. [ABSTRACT FROM AUTHOR]

Increased intracellular iron spurs mitochondrial biogenesis and respiration to satisfy high-energy demand during osteoclast differentiation and bone-resorbing activities. Transferrin receptor 1 (Tfr1) mediates cellular iron uptake through endocytosis of iron-loaded transferrin, and its expression increases during osteoclast differentiation. Nonetheless, the precise functions of Tfr1 and Tfr1-mediated iron uptake in osteoclast biology and skeletal homeostasis remain incompletely understood. To investigate the role of Tfr1 in osteoclast lineage cells in vivo and in vitro, we crossed Tfrc (encoding Tfr1)-floxed mice with Lyz2 (LysM)-Cre and Cathepsin K (Ctsk)-Cre mice to generate Tfrc conditional knockout mice in myeloid osteoclast precursors (Tfr1ΔLysM) or differentiated osteoclasts (Tfr1ΔLysM), respectively. Skeletal phenotyping by µCT and histology unveiled a significant increase in trabecular bone mass with normal osteoclast number in long bones of 10-week-old young and 6-month-old adult female but not male Tfr1ΔLysM mice. Although high trabecular bone volume in long bones was observed in both male and female Tfr1ΔLysM mice, this phenotype was more pronounced in female knockout mice. Consistent with this gender-dependent phenomena, estrogen deficiency induced by ovariectomy decreased trabecular bone mass in Tfr1ΔLysM mice. Mechanistically, disruption of Tfr1 expression attenuated mitochondrial metabolism and cytoskeletal organization in mature osteoclasts in vitro by attenuating mitochondrial respiration and activation of the Src-Rac1-WAVE regulatory complex axis, respectively, leading to decreased bone resorption with little impact on osteoclast differentiation. These results indicate that Tfr1-mediated iron uptake is specifically required for osteoclast function and is indispensable for bone remodeling in a gender-dependent manner. [ABSTRACT FROM AUTHOR]

• The role of Ser55 phosphorylation in enamelin was characterized in mice by eliminating or mimicking the phosphorylation state in vivo by substituting Ser into Ala or Asp using CRISPR-Cas9 approach. • Asp substitution partially mimicked the phosphorylation state of Ser55 and showed less disturbance to amelogenesis compared to Ala substitution. • Proteomic profiling suggests that the phosphorylation of Ser55 in enamelin is associated with enamel matrix biosynthetic and metabolic process. • These findings provide novel insights for understanding the biological role of oligo-phosphorylation in enamel matrix proteins. Amelogenesis imperfecta (AI) is an inherited developmental enamel defect affecting tooth masticatory function, esthetic appearance, and the well-being of patients. As one of the major enamel matrix proteins (EMPs), enamelin (ENAM) has three serines located in Ser-x-Glu (S-x-E) motifs, which are potential phosphorylation sites for the Golgi casein kinase FAM20C. Defects in FAM20C have similarly been associated with AI. In our previous study of Enam Rgsc514 mice, the Glu57 in the S55-X56-E57 motif was mutated into Gly, which was expected to cause a phosphorylation failure of Ser55 because Ser55 cannot be recognized by FAM20C. The severe enamel defects in ENAMRgsc514 mice reminiscent of Enam -knockout mouse enamel suggested a potentially important role of Ser55 phosphorylation in ENAM function. However, the enamel defects and ENAM dysfunction may also be attributed to distinct physicochemical differences between Glu57 and Gly57. To clarify the significance of Ser55 phosphorylation to ENAM function, we generated two lines of Enam knock-in mice using CRISPR-Cas9 method to eliminate or mimic the phosphorylation state of Ser55 by substituting it with Ala55 or Asp55 (designated as S55A or S55D), respectively. The teeth of 6-day or 4-week-old mice were subjected to histology, micro-CT, SEM, TEM, immunohistochemistry, and mass spectrometry analyses to characterize the morphological, microstructural and proteomic changes in ameloblasts, enamel matrix and enamel rods. Our results showed that the enamel formation and EMP expression in S55D heterozygotes (Het) were less disturbed than those in S55A heterozygotes, while both homozygotes (Homo) had no mature enamel formation. Proteomic analysis revealed alterations of enamel matrix biosynthetic and mineralization processes in S55A Hets. Our present findings indicate that Asp55 substitution partially mimics the phosphorylation state of Ser55 in ENAM. Ser55 phosphorylation is essential for ENAM function during amelogenesis. [ABSTRACT FROM AUTHOR]

Industrial energy systems, such as in the petrochemical and iron energy-intensive industries, are a utility hub to convert fuels into various forms of energy and then supply them to satisfy the energy requirements from process units. In conventional industrial energy systems design, the forms and quantity of energy required from process units are generally considered as deterministic. As a result, the optimization boundary of energy systems design is reduced in advance, leading to sub-optimal design results. In this study, energy interchangeability in process units is introduced to relax the bounds of energy requirements. These energy interchangeabilities can lead to different distribution patterns of energy requirement. In order to quantify the energy interchangeability of process units, energy consumption relaxation models (ECRMs) are presented. Two kinds of energy interchangeabilities are considered and investigated, one is the heat-based energy interchange between fuels and steam, another is the work-based energy interchange between electricity and steam. A candidate superstructure of industrial energy systems and a mixed integer nonlinear programming (MINLP) framework are then reformulated to conduct design optimization of the energy system to minimize the total annual cost (TAC). Lastly, the proposed method and MINLP optimization framework are applied to a large-scale refinery site for the design optimization of the energy system. Compared with the conventional optimized design, the TAC and annual CO 2 emission are decreased by 10.96% and 19,845 tons, respectively. [Display omitted] • Integration design of industrial energy systems and process units is investigated. • Energy interchangeability is introduced to relax the energy requirements. • Energy consumption relaxation models are presented for process units. • A MINLP mathematical framework is formulated for the design optimization. • TAC of the energy system in this case is decreased by 10.96%. [ABSTRACT FROM AUTHOR]

Objectives: In this study, we attempted to define the precise window of time for molar root elongation using a gain‐of‐function mutation of β‐catenin model. Materials and Methods: Both the control and constitutively activated β‐catenin (CA‐β‐cat) mice received a one‐time tamoxifen administration (for activation of β‐catenin at newborn, postnatal day 3, or 5, or 7, or 9) and were harvested at the same stage of P21. Multiple approaches were used to define the window of time of postnatal tooth root formation. Results: In the early activation groups (tamoxifen induction at newborn, or P3 or P5), there was a lack of molar root elongation in the CA‐β‐cat mice. When induced at P7, the root length was slightly reduced at P21. However, the root length was essentially the same as that in the control when β‐cat activated at P9. This study indicates that root elongation occurs in a narrow time of window, which is highly sensitive to a change of β‐catenin levels. Molecular studies showed a drastic decrease in the levels of nuclear factor I‐C (NFIC) and osterix (OSX), plus sharp reductions of odontoblast differentiation markers, including Nestin, dentin sialoprotein (DSP), and dentin matrix protein 1 (DMP1) at both mRNA and protein levels. Conclusions: Murine molar root elongation is precisely regulated by the Wnt/β‐catenin signaling within a narrow window of time (newborn to day 5). [ABSTRACT FROM AUTHOR]

The hedgehog (Hh) signaling pathway plays an essential role in both tissue development and homeostasis. Glioma‐associated oncogene homolog 1 (Gli1) is one of the vital transcriptional factors as well as the direct target gene in the Hh signaling pathway. The cells expressing the Gli1 gene (Gli1+ cells) have been identified as mesenchymal stem cells (MSCs) that are responsible for various tissue developments, homeostasis, and injury repair. This review outlines some recent discoveries on the crucial roles of Gli1+ MSCs in the development and homeostasis of varieties of hard and soft tissues. [ABSTRACT FROM AUTHOR]

Background: Chronic hepatitis B (CHB) and fatty liver (FL) are common, natural history data on concurrent FL and CHB (FL-CHB) are limited. This study aimed to evaluate the effect of FL on cirrhosis, hepatocellular carcinoma (HCC), and hepatitis B surface antigen (HBsAg) seroclearance incidence in CHB patients.Methods: In a retrospective cohort study of 6786 adult CHB patients, we used propensity score matching (PSM) to balance the FL-CHB and non-FL CHB groups. Kaplan-Meier methods were used to compare cumulative cirrhosis, HCC, and HBsAg seroclearance rates between subgroups.Results: Before PSM, compared to non-FL CHB, FL-CHB patients had lower 10-year cumulative rates of cirrhosis, HCC, and a higher HBsAg seroclearance rate. Similar results were found in the matched FL-CHB and non-FL CHB patients, as well as in the antiviral-treated PSM cohort. Cox proportional hazards model indicated FL to remain significantly and strongly associated with lower risk of cirrhosis and HCC (hazard ratio [HR], 0.19 [95% confidence interval {CI}, .12-.33], P < .001 and HR, 0.21 [95% CI, .09-.51], P = .001, respectively) in antiviral-treated patients but not in untreated patients.Conclusions: FL was significantly associated with lower cirrhosis and HCC risk and higher HBsAg seroclearance. Further studies are needed to confirm our funding and investigate the mechanisms underlying the impact of FL on CHB. [ABSTRACT FROM AUTHOR]

Abstract: Leishmania spp. protozoa are obligate intracellular parasites that replicate in macrophages during mammalian infection. Efficient phagocytosis and survival in macrophages are important determinants of parasite virulence. Macrophage lines differ dramatically in their ability to sustain intracellular Leishmania infantum chagasi (Lic). We report that the U937 monocytic cell line supported the intracellular replication and cell-to-cell spread of Lic during 72 h after parasite addition, whereas primary human monocyte-derived macrophages (MDMs) did not. Electron microscopy and live cell imaging illustrated that Lic promastigotes anchored to MDMs via their anterior ends and were engulfed through symmetrical pseudopods. In contrast, U937 cells bound Lic in diverse orientations, and extended membrane lamellae to reorient and internalize parasites through coiling phagocytosis. Lic associated tightly with the parasitophorous vacuole (PV) membrane in both cell types. PVs fused with LAMP-1-expressing compartments 24 h after phagocytosis by MDMs, whereas U937 cell PVs remained LAMP-1 negative. The expression of one phagocytic receptor (CR3) was higher in MDMs than U937 cells, leading us to speculate that parasite uptake proceeds through dissimilar pathways between these cells. We hypothesize that the mechanism of phagocytosis differs between primary versus immortalized human macrophage cells, with corresponding differences in the subsequent intracellular fate of the parasite. [Copyright &y& Elsevier]

The transcription factor Osterix (Osx) is required for osteoblast differentiation and bone formation during embryonic development, but it is not known whether Osx has an essential function in postnatal bone growth and in bone homeostasis. Conditional deletion of Osx at several time points postnatally revealed that Osx was essential for osteoblast differentiation and new bone formation in growing and adult bones. Additionally. inactivation of Osx in bones severely disrupted the maturation, morphology. and function of osteocytes. These findings identify Osx as having an essential role in the cell-specific genetic program of osteocytes. Interestingly, Osx inactivation also led to the massive accumulation of unresorbed calcified cartilage in a large area below the growth plate of endochondral bones. This specific area was also marked by an unanticipated almost complete lack of bone marrow cells and a marked decrease in the density and size of osteoclasts. This diminished density of osteoclasts could contribute to the lack of resorption of mineralized cartilage. In addition, we speculate that the abnormally accumulated, mainly naked cartilage represents an unfavorable substrate for osteoclasts. Our study identifies Osx as an essential multifunctional player in postnatal bone growth and homeostasis. [ABSTRACT FROM AUTHOR]

Pax9 and Msx1 encode transcription factors that are known to be essential for the switch in odontogenic potential from the epithelium to the mesenchyme. Multiple lines of evidence suggest that these molecules play an important role in the maintenance of mesenchymal Bmp4 expression, which ultimately drives morphogenesis of the dental organ. Here we demonstrate that Pax9 is able to directly regulate Msx1 expression and interact with Msx1 at the protein level to enhance its ability to transactivate Msx1 and Bmp4 expression during tooth development. In addition, we tested how a missense mutation (T62C) in the paired domain of PAX9 that is responsible for human tooth agenesis (1) affects its functions. Our data indicate that although the mutant Pax9 protein (L21P) can bind to the Msx1 protein, it fails to transactivate the Msx1 and Bmp4 promoter, presumably because of its inability to bind cognate paired domain recognition sequences. In addition, synergistic transcriptional activation of the Bmp4 promoter was lost with coexpression of mutant Pax9 and wild-type Msx1. This suggests that Pax9 is critical for the regulation of Bmp4 expression through its paired domain rather than Msx1. Our findings demonstrate the partnership of Pax9 and Msx1 in a signaling pathway that involves Bmp4. Furthermore, the regulation of Bmp4 expression by the interaction of Pax9 with Msx1 at the level of transcription and through formation of a protein complex determines the fate of the transition from bud to cap stage during tooth development. [ABSTRACT FROM AUTHOR]

Floral closure may be induced by pollination and various other factors, but is rarely studied comprehensively. Different kinds of floral closure should have various effects on reproductive fitness of plants. Two contrasting types of floral closure were observed in the flowers of Gentiana straminea Maxim. in the eastern Qinghai-Tibetan Plateau. The first type occurred prior to pollination during both gender phases, in response mainly to decreasing air temperatures. Flowers closed when decreasing temperatures approached 20°C and subsequently began to reopen the following day during mid-morning when air temperatures warmed to approximately 13–15 °C. This kind of floral closure can protect pollen grains on either stamens or stigmas, increasing fitness of both male and female. Following pollination, permanent floral closure occurred, although there was a delay between the dates of pollination and permanent closure, during which flowers continued to show temporary closure in response to low temperature episodes. The time required for permanent, pollination-induced closure varied according to the age of the gender phase, including a prolonged time before closure if pollination occurred early in the female phase. The retaining of permanent closed flowers increased both approaching (to inflorescences) and visiting (to unpollinated flowers) frequencies of individual plants when with fewer open flowers and the persisting corolla is further beneficial for seed sets of these pollinated flowers. Thus, two separate types of floral closure, one in response to environmental cues and the other in response to the age of each gender stage, appeared to have a strong influence on reproductive fitness in this species. These results revealed a different adaptive strategy of alpine plants in the sexual reproduction assurance in addition to the well-known elevated floral longevity, dominant role of more effective pollinators and increased reproduction allocation in the arid habitats. [ABSTRACT FROM AUTHOR]

To investigate the effect of isolated hydrostatic pressure on proteoglycan metabolism in chondrocytes. Bovine articular chondrocytes cultured in agarose gels were subjected to 5 MPa hydrostatic pressure for 4 hours in either a static or a pulsatile (1 Hz) mode, and changes in glycosaminoglycan (GAG) synthesis, hydrodynamic size, and aggregation properties of proteoglycans and aggrecan messenger RNA (mRNA) levels were determined. The application of 5 MPa static pressure caused a significant increase in GAG synthesis of 11% (P < 0.05). Column chromatography showed that this increase in GAG synthesis was associated with large proteoglycans. In addition, semiquantitative reverse transcriptase–polymerase chain reaction showed a 4-fold increase in levels of aggrecan mRNA (P < 0.01). Hydrostatic pressure in isolation, which does not cause cell deformation, can affect proteoglycan metabolism in chondrocytes cultured in agarose gels, indicating an important role of hydrostatic pressure in the regulation of extracellular matrix turnover in articular cartilage. [ABSTRACT FROM AUTHOR]

Background: Chronic hepatitis B (CHB) can progress to cirrhosis, but there are limited noninvasive tools available to estimate cirrhosis risk, including in patients receiving antiviral therapy. This study developed and validated a simple model to assess risk in CHB patients.Methods: The derivation cohort included 3000 CHB patients from 6 centers in the United States, with 52.60% receiving antiviral therapy. External validation was performed for 4552 CHB individuals from similar cohorts in Taiwan, with 21.27% receiving therapy. Cox proportional hazards regression analyses were used to screen predictors and develop the risk score for cirrhosis. Areas under receiver operating characteristic curves (AUROCs) were calculated for predictive value.Results: Sex, age, diabetes, antiviral treatment status/duration, hepatitis B e-antigen, and baseline alanine aminotransferase/aspartate aminotransferase levels were significantly associated with increased cirrhosis risk. A 13-point risk score was developed based on these predictors. The AUROCs for predicting cirrhosis risk were 0.82 at 3 years, 0.85 at 5 years, and 0.89 at 10 years in the derivation cohort, and 0.82, 0.79, and 0.77 in the validation cohort, respectively.Conclusions: We developed and validated a simple cirrhosis prediction model with an independent external cohort that can be applied to both treatment-naive and treatment-experienced CHB patients in diverse settings and locations. [ABSTRACT FROM AUTHOR]

Background: The formation of supernumerary teeth is an excellent model for studying the molecular mechanisms that control stem/progenitor cell homeostasis needed to generate a renewable source of replacement cells and tissues. Although multiple growth factors and transcriptional factors have been associated with supernumerary tooth formation, the regulatory inputs of extracellular matrix in this regenerative process remains poorly understood. Results: In this study, we present evidence that disrupting glycosaminoglycans (GAGs) in the dental epithelium of mice by inactivating FAM20B, a xylose kinase essential for GAG assembly, leads to supernumerary tooth formation in a pattern reminiscent of replacement teeth. The dental epithelial GAGs confine murine tooth number by restricting the homeostasis of Sox2(+) dental epithelial stem/progenitor cells in a non-autonomous manner. FAM20B-catalyzed GAGs regulate the cell fate of dental lamina by restricting FGFR2b signaling at the initial stage of tooth development to maintain a subtle balance between the renewal and differentiation of Sox2(+) cells. At the later cap stage, WNT signaling functions as a relay cue to facilitate the supernumerary tooth formation. Conclusions: The novel mechanism we have characterized through which GAGs control the tooth number in mice may also be more broadly relevant for potentiating signaling interactions in other tissues during development and tissue homeostasis. [ABSTRACT FROM AUTHOR]

Background: Patients on oral antiviral (OAV) therapy remain at hepatocellular carcinoma (HCC) risk. Risk prediction tools distinguishing treated patients with residual HCC risk are limited. The aim of this study was to develop an accurate, precise, simple-to-use HCC risk score using routine clinical variables among a treated Asian cohort.Methods: Adult Asian chronic hepatitis B (CHB) patients on OAV were recruited from 25 centers in the United States and the Asia-Pacific region. Excluded persons were coinfected with hepatitis C, D, or human immunodeficiency virus, had HCC before or within 1 year of study entry, or their follow-up was <1 year. Patients were randomized to derivation and validation cohorts on a 2:1 ratio. Statistically significant predictors from multivariate modeling formed the Real-world Effectiveness from the Asia Pacific Rim Liver Consortium for HBV (REAL-B) score.Results: A total of 8048 patients were randomized to the derivation (n = 5365) or validation group (n = 2683). The REAL-B model included 7 variables (male gender, age, alcohol use, diabetes, baseline cirrhosis, platelet count, and alpha fetoprotein), and scores were categorized as follows: 0-3 low risk, 4-7 moderate risk, and 8-13 high risk. Area under receiver operating characteristics were >0.80 for HCC risk at 3, 5, and 10 years, and these were significantly higher than other risk models (p < .001).Conclusions: The REAL-B score provides 3 distinct risk categories for HCC development in Asian CHB patients on OAV guiding HCC surveillance strategy. [ABSTRACT FROM AUTHOR]

А quantitative analysis method to determine the total nitrogen content in monoammonium phosphate (MAP) fertilizer using visible-near infrared (Vis-NIR) spectroscopy and least squares support vector machine (LS-SVM) is proposed. Sample set partitioning based on the joint x–y distance (SPXY) was used to select the calibration set. Fourteen spectral pre-processing methods were then employed to deal with the spectral data including Savitzky–Golay (SG) smoothing, fi rst derivative (D1) and second derivative (D2) with SG smoothing, multiplicative scatter correction (MSC), standard normal variate (SNV), wavelet, and combination thereof. Next, the LS-SVM model with radial basis function kernel was established with the best pre-processing method, and its performance was compared with that of partial least squares (PLS) model. The results revealed LS-SVM calibration with the discrete wavelet transform provided the best prediction for total nitrogen content in MAP fertilizer, yielding R2, root mean square error of prediction (RMSEP), and ratio of performance to deviation (RPD) values of 0.91, 0.101, and 3.34, respectively. [ABSTRACT FROM AUTHOR]

Objectives: Dental implants are major treatment options for restoring teeth loss. Biological processes at the implant‐tissue interface are critical for implant osseointegration. Superior mechanical properties of the implant constitute a major challenge for traditional histological techniques. It is imperative to develop new technique to investigate the implant‐tissue interface. Materials and methods: Our laboratory developed the polyethylene glycol (PEG)‐associated solvent system (PEGASOS) tissue clearing method. By immersing samples into various chemical substances, bones and teeth could be turned to transparent with intact internal structures and endogenous fluorescence being preserved. We combined the PEGASOS tissue clearing method with transgenic mouse line and other labelling technique to investigate the angiogenesis and osteogenesis processes occurring at the implant‐bone interface. Results: Clearing treatment turned tissue highly transparent and implant could be directly visualized without sectioning. Implant, soft/hard tissues and fluorescent labels were simultaneously imaged in decalcified or non‐decalcified mouse mandible samples without disturbing their interfaces. Multi‐channel 3‐dimensional image stacks at high resolution were acquired and quantified. The processes of angiogenesis and osteogenesis surrounding titanium or stainless steel implants were investigated. Conclusions: Both titanium and stainless steel implants support angiogenesis at comparable levels. Successful osseointegration and calcium precipitation occurred only surrounding titanium, but not stainless steel implants. PEGASOS tissue clearing method provides a novel approach for investigating the interface between implants and hard tissue. [ABSTRACT FROM AUTHOR]

Highlights • Resveratrol suppressed TNFα-induced inflammatory cytokines produced by human dental pulp stem cells (DPSCs). • Resveratrol functioned through regulating the inhibitory autophagy-JNK signaling cascade in DPSCs. • Resveratrol might be beneficial to ameliorate pulpal damage in the acute phase of pulpal inflammation in vital pulp therapy. Abstract Objectives: To study the effects of polyphenol resveratrol on TNFα-induced inflammatory signaling as well as the underlying mechanism in human dental pulp stem cells (DPSCs). Materials and methods: Human DPSCs were cultured and treated by TNFα in the presence or absence of resveratrol. NF-κB and mitogen-activated protein kinase (MAPK) signaling pathways were analyzed by Western blotting and immunofluorescence staining. Interleukin 6 (IL6) and interleukin 8 (IL8) mRNA levels were analyzed by reverse transcription polymerase chain reaction. For the mechanistic study, autophagy was examined and further manipulated by gene silencing of Atg5 using siRNAs. Statistical analysis was performed by Student's t - test, and values of p < 0.05 were considered significant. Results: Upon TNFα treatments, neither degradation of IκBα nor the phosphorylation and nuclear translocation of p65 NF-κB were inhibited by resveratrol at different concentrations. In contrast, resveratrol dramatically inhibited TNFα-induced phosphorylation of c-Jun N-terminal kinase (JNK) MAPK. Furthermore, resveratrol activated autophagy, as evidenced by the accumulated autophagic puncta formed by lipid bound LC3B in resveratrol-treated cells. Intriguingly, both resveratrol and JNK inhibitor SP600125 suppressed TNFα-induced IL6 and IL8 mRNA expression (P < 0.05). Silencing autophagy gene Atg5 led to the hyper-activation of JNK and augmented TNFα-induced IL6 and IL8 mRNA expression (P < 0.05). Conclusions: The results suggest that resveratrol suppresses TNFα-induced inflammatory cytokines expressed by DPSCs through regulating the inhibitory autophagy-JNK signaling cascade. Resveratrol might be beneficial to ameliorate pulpal damage during the acute phase of inflammation in vital pulp therapy. [ABSTRACT FROM AUTHOR]