bluuebunny/crossref_metadata_2025_split huggingface dataset:
Structure–function analysis of the LytM domain of EnvC, an activator of cell wall remodelling at the Escherichia coli division site
Authors: Peters Nick T. , Morlot Cécile , Yang Desirée C. , Uehara Tsuyoshi , Vernet Thierry , Bernhardt Thomas G. 
Published: June 2013

SummaryProteins with LytM (Peptidase_M23) domains are broadly distributed in bacteria and have been implicated in a variety of important processes, including cell division and cell‐shape determination. Most LytM‐like proteins that have been structurally and/or biochemically characterized are metallo‐endopeptidases that cleave cross‐links in the peptidoglycan (PG) cell wall matrix. Notable exceptions are the Escherichia coli cell division proteins EnvC and NlpD. These LytM factors are not hydrolases themselves, but instead serve as activators that stimulate PG cleavage by target enzymes called amidases to promote cell separation. Here we report the structure of the LytM domain from EnvC, the first structure of a LytM factor implicated in the regulation of PG hydrolysis. As expected, the fold is highly similar to that of other LytM proteins. However, consistent with its role as a regulator, the active‐site region is degenerate and lacks a catalytic metal ion. Importantly, genetic analysis indicates that residues in and around this degenerate active site are critical for amidase activation in vivo and in vitro. Thus, in the regulatory LytM factors, the apparent substrate binding pocket conserved in active metallo‐endopeptidases has been adapted to control PG hydrolysis by another set of enzymes.
Proteomic profiling of developing cotton fibers from wild and domesticatedGossypium barbadense
Authors: Hu Guanjing , Koh Jin , Yoo Mi‐Jeong , Grupp Kara , Chen Sixue , Wendel Jonathan F. 
Published: June 2013

SummaryPima cotton (Gossypium barbadense) is widely cultivated because of its long, strong seed trichomes (‘fibers’) used for premium textiles. These agronomically advanced fibers were derived following domestication and thousands of years of human‐mediated crop improvement. To gain an insight into fiber development and evolution, we conducted comparative proteomic and transcriptomic profiling of developing fiber from an elite cultivar and a wild accession.Analyses using isobaric tag for relative and absolute quantification (iTRAQ)LC‐MS/MStechnology identified 1317 proteins in fiber. Of these, 205 were differentially expressed across developmental stages, and 190 showed differential expression between wild and cultivated forms, 14.4% of the proteome sampled. Human selection may have shifted the timing of developmental modules, such that some occur earlier in domesticated than in wild cotton.A novel approach was used to detect possible biased expression of homoeologous copies of proteins. Results indicate a significant partitioning of duplicate gene expression at the protein level, but an approximately equal degree of bias for each of the two constituent genomes of allopolyploid cotton.Our results demonstrate the power of complementary transcriptomic and proteomic approaches for the study of the domestication process. They also provide a rich database for mining for functional analyses of cotton improvement or evolution.
Mesotherapy for local fat reduction
Authors: Jayasinghe S. , Guillot T. , Bissoon L. , Greenway F. 
Published: June 2013

SummaryMesotherapy, which is the injection of substances locally into mesodermally derived subcutaneous tissue, developed from empirical observations of a French physician in the 1950s. Although popular in Europe for many medical purposes, it is used for local cosmetic fat reduction in the United States. This paper reviews manuscripts indexed in PubMed/MEDLINE under ‘mesotherapy’, which pertains to local fat reduction. The history of lipolytic mesotherapy, the physiology of body fat distribution, the mechanism of action of different lipolytic stimulators and their increased efficacy in combination are reviewed. Mesotherapy falls into two categories. Lipolytic mesotherapy using lipolytic stimulators requires more frequent treatments as the fat cells are not destroyed and can refill over time. Ablative mesotherapy destroys fat cells with a detergent, causes inflammation and scarring from the fat necrosis, but requires fewer treatments. The historic and empiric mixing of sodium channel blocking local anaesthetics in mesotherapy solutions inhibits the intended lipolysis. Major mesotherapy safety concerns include injection site infections from poor sterile technique. Cosmetic mesotherapy directs the area from which fat is lost to improve self‐image. Studies were of relatively small number, many with limited sample sizes. Future research should be directed towards achieving a Food and Drug Administration indication rather than continuing expansion of off‐label use.
Hypoxia induces osteogenesis in rabbit adipose‐derived stem cells overexpressing bone morphogenic protein‐2
Authors: Xu L , Sun X , Cao K , Wu Y , Zou D , Liu Y , Zhang X , Zhang X , Wang G , Huang Q , Jiang X 
Published: June 2013

ObjectiveHypoxic culture potentiates mesenchymal stem cells (MSCs) to survive and secrete various growth factors. Genetically modified stem cells overexpressing bone morphogenic protein‐2 (BMP‐2) demonstrate strong osteogenic ability. Hence, we investigated the coeffect of hypoxic culture conditions and BMP‐2 overexpression on the osteogenic ability of rabbit adipose‐derived stem cells (rASCs) in vitro.Materials and MethodsRabbit adipose‐derived stem cells with or without adenoviral‐BMP‐2 transduction were cultured in hypoxic (1%) and normoxic (21%) conditions. Cell viability, attachment, and proliferation were compared. Real‐time PCR amplification of osteogenic and angiogenic genes including alkaline phosphatase (ALP), osteocalcin (OCN), HIF‐1α, and vascular endothelial growth factor (VEGF) was performed. Moreover, ALP activity, immunofluorescent staining of OCN, and mineralization assay by alizarin red S quantification and von Kossa staining were conducted.ResultsCells under hypoxic conditions attached better within 12 h and proliferated faster. While BMP‐2 overexpression and hypoxic condition separately elevated the transcription of key osteogenic and angiogenic genes, a cooperative effect was observed to enhance the upregulation of osteogenic as well as angiogenic genes. Identical changes were observed in ALP activity, immunofluorescent staining of OCN, and mineralization assay.ConclusionsHypoxic culture can enhance the osteogenic ability of BMP‐2 gene‐modified rASCs, which provides a strategy to improve the osteogenesis of rASCs for in vivo bone regeneration.
Alendronate impairs epithelial adhesion, differentiation and proliferation in human oral mucosa
Authors: Donetti E , Gualerzi A , Sardella A , Lodi G , Carrassi A , Sforza C 
Published: June 2013

ObjectiveThis study aimed at evaluating from a morphological point of view the effects of alendronate (ALN), a widely used nitrogen‐containing bisphosphonate for the chronic treatment of osteoporosis, on the oral epithelium of healthy keratinized human oral mucosa. Bisphosphonate‐related osteonecrosis of the jaw is a well‐known severe consequence, but the effects during chronic therapy on the oral soft tissues are still matter of debate.Materials and MethodsSix women over 60 year‐old undergoing treatment of osteoporosis with 70 mg per week of oral ALN (lasting at least 2 years) were recruited and compared with a gender and age‐matched group (n = 6). Proliferation, apoptosis, intercellular adhesion and terminal differentiation (TD) were investigated by immunofluorescence. In parallel, ultrastructural analysis was carried out.ResultsBy immunofluorescence, a statistically significant decrease in keratinocyte proliferation was detected in the oral epithelium of the ALN group without any sign of apoptosis, but accompanied by a reduction in desmoglein 1 and keratin 10 expressions. In the uppermost layers of the oral epithelium of the ALN group, thin desmosomes were visible by transmission electron microscopy.ConclusionOur results show that epithelial adhesion, TD and proliferation are affected by ALN therapeutic doses in clinically healthy human oral mucosa.
Variability and consistency in lung inflammatory responses to particles with a geogenic origin
Authors: Zosky Graeme R. , Boylen Catherine E. , Wong Russell S. , Smirk Michael N. , Gutiérrez Lucia , Woodward Robert C. , Siah W. Shan , Devine Brian , Maley Fiona , Cook Angus 
Published: June 2013

AbstractBackground and objectiveParticulate matter <10 μm (PM10) is well recognized as being an important driver of respiratory health; however, the impact of PM10 of geogenic origin on inflammatory responses in the lung is poorly understood. This study aimed to assess the lung inflammatory response to community sampled geogenic PM10.MethodsThis was achieved by collecting earth material from two regional communities in Western Australia (Kalgoorlie‐Boulder and Newman), extracting the PM10 fraction and exposing mice by intranasal instillation to these particles. The physicochemical characteristics of the particles were assessed and lung inflammatory responses were compared to control particles. The primary outcomes were cellular influx and cytokine production in the lungs of the exposed mice.ResultsThe physical and chemical characteristics of the PM10 from Kalgoorlie and Newman differed with the latter having a higher concentration of Fe and a larger median diameter. Control particles (2.5 μm polystyrene) caused a significant influx of inflammatory cells (neutrophils) with little production of proinflammatory cytokines. In contrast, the geogenic particles induced the production of MIP‐2, IL‐6 and a significant influx of neutrophils. Qualitatively, the response following exposure to particles from Kalgoorlie and Newman were consistent; however, the magnitude of the response was substantially higher in the mice exposed to particles from Newman.ConclusionsThe unique physicochemical characteristics of geogenic particles induced a proinflammatory response in the lung. These data suggest that particle composition should be considered when setting community standards for PM exposure, particularly in areas exposed to high geogenic particulate loads.
Comparative study of hair follicle morphology in eight mammalian species and humans
Authors: Mangelsdorf Susanne , Vergou Theognosia , Sterry Wolfram , Lademann Juergen , Patzelt Alexa 
Published: June 2013

BackgroundThe objective of the present study was the investigation of hair follicle morphology in eight mammalian species in order to evaluate the species‐specific contribution of hair follicles to skin penetration particularly with regard to the utilization of the different animal species as skin models for human skin.MethodsCyanoacrylate skin surface biopsy method (CSSB), light microscopy and also digital photography were used for the measurements of hair follicle morphology.ResultsThe results revealed species‐specific differences regarding the pattern of hair follicle distribution and also differences with regard to hair follicle parameters and characteristics. The results also showed that hair follicles generally possess enormous reservoir capacities, regarding the follicular volume. In all examined species, hair follicles reached at least one‐fifth of stratum corneum storage capacity. The results were compared with human data obtained in a previous study.ConclusionWith regard to hair follicle morphology and skin structure, the porcine skin seems to be the most appropriate skin model for human skin analog to previous investigations, whereas the skin of dog, cat, and rabbit showed the most significant differences.
Regulation of miRNAs in human skeletal muscle following acute endurance exercise and short‐term endurance training
Authors: Russell Aaron P. , Lamon Severine , Boon Hanneke , Wada Shogo , Güller Isabelle , Brown Erin L. , Chibalin Alexander V. , Zierath Juleen R. , Snow Rod J. , Stepto Nigel , Wadley Glenn D. , Akimoto Takayuki 
Published: June 2013

Key points
The discovery of microRNAs (miRNAs) has established new mechanisms that control health, but little is known about the regulation of skeletal muscle miRNAs in response to exercise.
This study investigated components of the miRNA biogenesis pathway (Drosha, Dicer and Exportin‐5), muscle enriched miRNAs, (miR‐1, ‐133a, ‐133b and 206), and several miRNAs dysregulated in muscle myopathies, and showed that 3 h following an acute exercise bout, Drosha, Dicer and Exportin‐5, as well as miR‐1, ‐133a, ‐133‐b and miR‐181a were all increased, while miR‐9, ‐23a, ‐23b and ‐31 were decreased.
Short‐term training increased miR‐1 and miR‐29b, while miR‐31 remained decreased.
Negative correlations were observed between miR‐9 and HDAC4 protein, miR‐31 and HDAC4 protein and between miR‐31 and NRF1 protein, 3 h after exercise.
miR‐31 binding to the HDAC4 and NRF1 3′ untranslated region (UTR) reduced luciferase reporter activity.
Exercise rapidly and transiently regulates several miRNA species potentially involved in the regulation of skeletal muscle regeneration, gene transcription and mitochondrial biogenesis.
Abstract  The identification of microRNAs (miRNAs) has established new mechanisms that control skeletal muscle adaptation to exercise. The present study investigated the mRNA regulation of components of the miRNA biogenesis pathway (Drosha, Dicer and Exportin‐5), muscle enriched miRNAs, (miR‐1, ‐133a, ‐133b and ‐206), and several miRNAs dysregulated in muscle myopathies (miR‐9, ‐23, ‐29, ‐31 and ‐181). Measurements were made in muscle biopsies from nine healthy untrained males at rest, 3 h following an acute bout of moderate‐intensity endurance cycling and following 10 days of endurance training. Bioinformatics analysis was used to predict potential miRNA targets. In the 3 h period following the acute exercise bout, Drosha, Dicer and Exportin‐5, as well as miR‐1, ‐133a, ‐133‐b and ‐181a were all increased. In contrast miR‐9, ‐23a, ‐23b and ‐31 were decreased. Short‐term training increased miR‐1 and ‐29b, while miR‐31 remained decreased. Negative correlations were observed between miR‐9 and HDAC4 protein (r=−0.71; P= 0.04), miR‐31 and HDAC4 protein (r =−0.87; P= 0.026) and miR‐31 and NRF1 protein (r =−0.77; P= 0.01) 3 h following exercise. miR‐31 binding to the HDAC4 and NRF1 3′ untranslated region (UTR) reduced luciferase reporter activity. Exercise rapidly and transiently regulates several miRNA species in muscle. Several of these miRNAs may be involved in the regulation of skeletal muscle regeneration, gene transcription and mitochondrial biogenesis. Identifying endurance exercise‐mediated stress signals regulating skeletal muscle miRNAs, as well as validating their targets and regulatory pathways post exercise, will advance our understanding of their potential role/s in human health.
Dosimetric accuracy of proton therapy for chordoma patients with titanium implants
Authors: Verburg Joost M. , Seco Joao 
Published: June 2013

Purpose:To investigate dosimetric errors in proton therapy treatment planning due to titanium implants, and to determine how these affect postoperative passively scattered proton therapy for chordoma patients with orthopedic hardware.Methods:The presence of titanium hardware near the tumor may affect the dosimetric accuracy of proton therapy. Artifacts in the computed tomography (CT) scan can cause errors in the proton stopping powers used for dose calculation, which are derived from CT numbers. Also, clinical dose calculation algorithms may not accurately simulate proton beam transport through the implants, which have very different properties as compared to human tissue. The authors first evaluated the impact of these two main issues. Dose errors introduced by metal artifacts were studied using phantoms with and without titanium inserts, and patient scans on which a metal artifact reduction method was applied. Pencil‐beam dose calculations were compared to models of nuclear interactions in titanium and Monte Carlo simulations. Then, to assess the overall impact on treatment plans for chordoma, the authors compared the original clinical treatment plans to recalculated dose distributions employing both metal artifact reduction and Monte Carlo methods.Results:Dose recalculations of clinical proton fields showed that metal artifacts cause range errors up to 6 mm distal to regions affected by CT artifacts. Monte Carlo simulations revealed dose differences >10% in the high‐dose area, and range differences up to 10 mm. Since these errors are mostly local in nature, the large number of fields limits the impact on target coverage in the chordoma treatment plans to a small decrease of dose homogeneity.Conclusions:In the presence of titanium implants, CT metal artifacts and the approximations of pencil‐beam dose calculations cause considerable errors in proton dose calculation. The spatial distribution of the errors however limits the overall impact on passively scattered proton therapy for chordoma.
Radiologic Responses in Cynomolgus Macaques for Assessing Tuberculosis Chemotherapy Regimens
Authors: Lin Philana Ling , Coleman Teresa , Carney Jonathan P. J. , Lopresti Brian J. , Tomko Jaime , Fillmore Dan , Dartois Veronique , Scanga Charles , Frye L. James , Janssen Christopher , Klein Edwin , Barry Clifton E. , Flynn JoAnne L. 
Published: June 2013

ABSTRACT

Trials to test new drugs currently in development against tuberculosis in humans are impractical. All animal models to prioritize new regimens are imperfect, but nonhuman primates (NHPs) infected with
Mycobacterium tuberculosis
develop active tuberculosis (TB) disease with a full spectrum of lesion types seen in humans. Serial 2-deoxy-2-[
18
F]-fluoro-
d
-glucose (FDG) positron emission tomography (PET) with computed tomography (CT) imaging was performed on cynomolgus macaques during infection and chemotherapy with individual agents or the four-drug combination therapy most widely used globally. The size and metabolic activity of lung granulomas varied among animals and even within a single animal during development of disease. Individual granulomas within untreated animals had highly local and independent outcomes, some progressing in size and FDG uptake, while others waned, illustrating the highly dynamic nature of active TB. At necropsy, even untreated animals were found to have a proportion of sterile lesions consistent with the dynamics of this infection. A more marked reduction in overall metabolic activity in the lungs (decreased FDG uptake) was associated with effective treatment. A reduction in the size of individual lesions correlated with a lower bacterial burden at necropsy. Isoniazid treatment was associated with a transient increase in metabolic activity in individual lesions, whereas a net reduction occurred in most lesions from rifampin-treated animals. Quadruple-drug therapy resulted in the highest decrease in FDG uptake. The findings of PET-CT imaging may provide an important early correlate of the efficacy of novel combinations of new drugs that can be directly translated to human clinical trials.
Determination of
In Vitro
Activities of Solithromycin at Different pHs and Its Intracellular Activity against Clinical Isolates of Neisseria gonorrhoeae from a Laboratory Collection
Authors: Mallegol Julia , Fernandes Prabhavathi , Seah Christine , Guyard Cyril , Melano Roberto G. 
Published: June 2013

ABSTRACT

We evaluated the activity of solithromycin against 196 clinical gonococcal isolates collected at the Public Health Ontario Laboratories, Toronto, Canada, including isolates with different levels of azithromycin resistance, as well as the role of pH in MIC determinations using pH-adjusted agar plates (pH range, 5.6 to 7.6).
In vitro
invasion assays were performed using monolayers of HeLa epithelial cells and clinical gonococci displaying different azithromycin MICs; infected cultures were treated with solithromycin, and its intracellular activity was determined by CFU assays after 3 and 20 h of exposure. Solithromycin displayed a MIC
50
and MIC
90
of 0.0625 and 0.125 μg/ml, respectively, making its activity at least 4-fold higher than that of azithromycin. Clinical isolates with elevated MICs for azithromycin (MICs of ≥2,048 μg/ml and 4 to 8 μg/ml) showed solithromycin MIC values of 8 and 0.5 μg/ml, respectively. In contrast to azithromycin, solithromycin MICs were not significantly affected by acidic pHs, suggesting more stability at lower pH. Moreover, when intracellular
Neisseria gonorrhoeae
isolates were incubated with solithromycin at 4 times, 1 times, and one-fourth of the MIC, the exposure to solithromycin resulted in the progressive loss of viability of most isolates over time. The intracellular activity of solithromycin, combined with the low MICs to this agent, indicates that it may be an attractive option for gonorrhea treatment if clinical trials in development reveal that this drug can be used safely in adult indications, especially when multidrug-resistant clinical isolates are now emerging.
Prototypical Recombinant Multi-Protease-Inhibitor-Resistant Infectious Molecular Clones of Human Immunodeficiency Virus Type 1
Authors: Varghese Vici , Mitsuya Yumi , Fessel W. Jeffrey , Liu Tommy F. , Melikian George L. , Katzenstein David A. , Schiffer Celia A. , Holmes Susan P. , Shafer Robert W. 
Published: June 2013

ABSTRACT
The many genetic manifestations of HIV-1 protease inhibitor (PI) resistance present challenges to research into the mechanisms of PI resistance and the assessment of new PIs. To address these challenges, we created a panel of recombinant multi-PI-resistant infectious molecular clones designed to represent the spectrum of clinically relevant multi-PI-resistant viruses. To assess the representativeness of this panel, we examined the sequences of the panel's viruses in the context of a correlation network of PI resistance amino acid substitutions in sequences from more than 10,000 patients. The panel of recombinant infectious molecular clones comprised 29 of 41 study-defined PI resistance amino acid substitutions and 23 of the 27 tightest amino acid substitution clusters. Based on their phenotypic properties, the clones were classified into four groups with increasing cross-resistance to the PIs most commonly used for salvage therapy: lopinavir (LPV), tipranavir (TPV), and darunavir (DRV). The panel of recombinant infectious molecular clones has been made available without restriction through the NIH AIDS Research and Reference Reagent Program. The public availability of the panel makes it possible to compare the inhibitory activities of different PIs with one another. The diversity of the panel and the high-level PI resistance of its clones suggest that investigational PIs active against the clones in this panel will retain antiviral activity against most if not all clinically relevant PI-resistant viruses.