Overexpression of protein kinase C-delta plays a crucial role in interleukin-6-producing pheochromocytoma presenting with acute inflammatory syndrome: a case report.
Pheochromocytomas are tumors that may produce a variety of substances in addition to catecholamines. To date, among several cases of systemic inflammatory syndrome associated with interleukin-6 (IL-6) secretion, IL-6-producing pheochromocytomas, have been reported. However, the mechanism underlying IL-6 oversecretion in these cases has not yet been clarified. This report describes a patient with pheochromocytoma who exhibited pyrexia and marked inflammatory signs including C-reactive protein elevation. The inflammatory symptoms were easily controlled by the administration of loxoprofen, a nonsteroidal anti-inflammatory drug. The plasma concentration of IL-6 and 11-d-TXB(2), a stable metabolite of thromboxane A(2) (TXA(2)), were significantly elevated in parallel with an elevation of norepinephrine in the samples obtained by selective venous sampling. A left adrenalectomy was performed, and the acute inflammatory symptoms naturally diminished without loxoprofen. Cultured tumor cells obtained from the resected specimen spontaneously released IL-6, and indomethacin inhibited the IL-6 release. According to a cDNA microarray analysis, mRNA of protein kinase C-delta (PKC-delta), prostaglandin D synthase, and arachidonate release-relating enzymes were significantly overexpressed in the tumor tissue in comparison to the adjacent nontumor tissue. The constitutive phosphorylation of PKC-delta was observed in the tumor tissue. These results strongly suggest that the systemic inflammatory syndrome in IL-6-producing pheochromocytoma, at least in part, is caused by the overexpression of PKC-delta, resulting in an excess of arachidonate derivatives such as prostaglandins.


Transcriptomic Profiling Reveals Chemokine CXCL1 as a Mediator for Neutrophil Recruitment Associated With Blood-Retinal Barrier Alteration in Diabetic Retinopathy.
Inflammation plays an important role in the pathogenesis of diabetic retinopathy (DR). To precisely define the inflammatory mediators, we examined the transcriptomic profile of human retinal endothelial cells exposed to advanced glycation end products, which revealed the neutrophil chemoattractant chemokine CXCL1 as one of the top genes upregulated. The effect of neutrophils in the alteration of the blood-retinal barrier (BRB) was further assessed in wild-type C57BL/6J mice intravitreally injected with recombinant CXCL1 as well as in streptozotocin-induced diabetic mice. Both intravitreally CXCL1-injected and diabetic animals showed significantly increased retinal vascular permeability, with significant increase in infiltration of neutrophils and monocytes in retinas and increased expression of chemokines and their receptors, proteases, and adhesion molecules. Treatment with Ly6G antibody for neutrophil depletion in both diabetic mice as well as CXCL1-injected animals showed significantly decreased retinal vascular permeability accompanied by decreased infiltration of neutrophils and monocytes and decreased expression of cytokines and proteases. CXCL1 level was significantly increased in the serum samples of patients with DR compared with samples of those without diabetes. These data reveal a novel mechanism by which the chemokine CXCL1, through neutrophil recruitment, alters the BRB in DR and, thus, serves as a potential novel therapeutic target. Intravitreal CXCL1 injection and diabetes result in increased retinal vascular permeability with neutrophil and monocyte recruitment. Ly6G antibody treatment for neutrophil depletion in both animal models showed decreased retinal permeability and decreased cytokine expression. CXCL1 is produced by retinal endothelial cells, pericytes, and astrocytes. CXCL1 level is significantly increased in serum samples of patients with diabetic retinopathy. CXCL1, through neutrophil recruitment, alters the blood-retinal barrier in diabetic retinopathy and, thus, may be used as a therapeutic target.


Tumor-specific near-infrared nanobody probe rapidly labels tumors in an orthotopic mouse model of pancreatic cancer.
Nanobodies, derived from camelid antibodies made of only heavy chains, are the smallest, biologic, antigen-binding fragments (~15kDa) with faster pharmacokinetics and better tumor penetration efficiency than standard antibodies. The present study evaluates the efficacy of a fluorescent, anti-carcinoembryonic antigen (CEA) nanobody for rapid tumor labeling in an orthotopic mouse model of pancreatic cancer. Anti-CEA or control nanobodies were conjugated with the near-infrared fluorophore IRDye 800CW. Fragments of BxPC-3 (high-CEA expressing) or MiaPACA-2 (low-CEA expressing) human pancreatic cancer cell lines were orthotopically implanted into the pancreatic tail of nude mice. After tumors reached 7 to 10 mm in size, 2 nmol anti-CEA or control nanobody-IRDye800CW were injected intravenously. Mice were imaged at various time points hours post-injection. Anti-CEA nanobodies clearly labeled BxPC3 orthotopic pancreatic tumors 3 hours after injection. The signal was present as early as 15 minutes after injection and was robust at 1 to 3 hours after injection with a tumor-to-background ratio of 2.66. In contrast, there was very low accumulation in the low CEA-expressing, MiaPACA2 pancreatic orthotopic tumors. The fluorophore-conjugated nanobody was specific for CEA-expressing tumors, while the control nanobody did not show any tumor-specific signal. Both nanobodies had strong kidney uptake as expected for small-molecule probes. The fluorescence signal was detectable using 2 clinical, Food and Drug Administration-approved, 800 nm imaging devices as well as small animal imaging systems. This anti-CEA, nanobody-based, fluorescent probe labeled pancreatic orthotopic tumors within 15 minutes of intravenous injection. Fluorescent anti-CEA nanobodies have labeling kinetics that approach the speed of nonspecific dyes such as indocyanine green but with the specificity of antibodies. The use of fluorescently-labeled, intact antibodies leads to a labeling delay of 48 to 96 hours between probe administration and the necessarily delayed time of operation, which can be avoided with nanobodies. The kinetics of a nanobody-based probe makes it a practical agent for same-day, patient administration and fluorescence-guided surgery.


Comparison of the Clinical and Genotypic Characteristics of Uropathogenic Escherichia coli Strains According to Sex in Korea.
In this study, we compared the microbiological, genotypic, and antibiotic resistance characteristics of uropathogenic Escherichia coli (UPEC) strains in patients with pyelonephritis in Korea according to sex based on data corresponding to the February 2015 to June 2018 period. Based on Escherichia coli phylogenetic group analysis, gene virulence detection, and subgroup analyses by sex, we observed that the antibiotic resistance percentages and proportions corresponding to extended-spectrum beta-lactamase producing UPEC were higher in males than in females. In addition, phylogenetic group B2 showed predominance in both the male and female groups, which further showed similar adhesion molecule distributions. Toxin-associated factors, hlyA and cnf1, were more common in males. In clinical presentations, urinary predisposing factors, complicated urinary tract infections (UTIs), concomitant bacteremia, and persistent fever were also more common with males. Although females and males showed UPEC genotypic differences, there were no differences between them with respect to poor outcomes. Persistent fever was associated with community-acquired infection and bacteremic UTI and relapsed UTI within 3 months was associated with urinary tract stone. In future, it will be necessary to conduct multicenter studies, involving more cases on UPEC to validate our results.


Hemiparesis in spontaneous spinal epidural haematoma: a potential stroke imitator.
Spontaneous spinal epidural haematoma (SSEH) is a rare condition that requires urgent surgical intervention in order to prevent permanent neurological deficit. SSEH commonly presents as a paraparesis or tetraparesis. SSEH presenting as a hemiparesis is less common and in such situations, it can be mistaken for a cerebrovascular accident (CVA). Thrombolytic or anticoagulant treatment for CVA can potentially worsen the neurological deficit. We report one such case of SSEH misdiagnosed as a CVA. Treatment with tissue plasminogen activator led to worsening of his condition. On a subsequent cervical spine MRI, an epidural haematoma extending from C3 to C5 was detected and treated with laminectomy and evacuation. Surgical intervention led to significant improvement from American Spinal Injury Association Scale (ASIA) B to ASIA E. Presence of clinical features such as Horner's syndrome, Brown-Sequard syndrome and the absence of cranial nerve palsies in acute hemiparesis are indicative of SSEH rather than CVA.


Continuum equations for magnetic and dielectric fluids with internal rotations.
Several authors have attempted with varying success to derive a complete set of basic equations for the motion of polar fluids having internal rotations and hence in a state of polarization disequilibrium. This work develops a complete set of governing equations derived on the basis of dynamic balance relationships with the dissipation function determined from thermodynamic consideration. The magnetization relaxation equation is thereby determined from requirement of positive entropy production along with a complete set of constitutive laws including antisymmetric terms of the total stress tensor. The analysis employs the Minkowski expression of electromagnetic momentum and assumes that the product of electromagnetic stress and velocity contributes to the energy balance on the same footing as contact stresses of pressure and viscous origin. The work refines the treatment of our earlier effort carrying out the analysis to first order in the ratio of fluid velocity to light speed throughout.


Development of a method for preparing cyclic nigerosylnigerose syrup and investigation of its value as a dietary fiber.
Cyclic nigerosylnigerose (CNN) syrup, containing 76% water-soluble dietary fiber, was prepared from starch on an industrial scale, using isoamylase, 6-α-glucosyltransferase, 3-α-isomaltosyltransferase, and cyclodextrin glucanotransferase. CNN syrup has a unique linkage pattern, consisting mainly of α-1,3 and α-1,6 glucoside linkages, and is characterized by its low weight average molecular weight (807) and moderate sweetness (relative sweetness = 25), unlike in well-known dietary fiber materials. The glass transition temperature of CNN is higher than that of the straight chain structures, maltotetraose and maltosyltrehalose. Even when 40% of normally added sucrose was replaced with CNN syrup, sponge cake puffed up sufficiently. The no observed adverse effect level for a single dose of CNN syrup was 0.88 and 0.89 g dry solid/kg body weight for men and women, respectively. The increase in blood glucose and insulin concentrations during consumption of CNN syrup was lower than that of glucose.


Relation between change in exercise capacity and change in blood amino acids in patients with chronic heart failure.
[Purpose] Although cardiac rehabilitation (CR) is recommended for patients with chronic heart failure (CHF), adequate exercise effect cannot be obtained in elderly patients. Administration of amino acids (AA) to CHF patients has been reported to improve exercise capacity, but the changes in AA composition in plasma before and after CR had not been reported. This study aimed to measure plasma levels of AA in CHF patients and compare with values of normal range. In addition the relationship between the change of exercise capacity and AA were examined. [Subjects and Methods] Twelve CHF patients (60% males, aged 68 ± 12 years) were studied. The correction between the rates of changes in exercise capacity parameters and in plasma AA levels was investigated. [Results] Anaerobic threshold (AT) and peak oxygen uptake (VO2) improved significantly after CR. The AA profile showed no specific pattern, and citrulline (Cit) was the amino acid showing a significant positive correlation with exercise capacity (∆Cit vs. ∆AT: r=0.602, ∆Cit vs. ∆AT-work rate (WR): r=0.681, ∆Cit vs. ∆VO2/WR: r=0.635). A tendency of positive correlation was observed between ∆Cit and ∆peak VO2 (r=0.456). [Conclusion] The AA profile showed no specific pattern, but a relationship between change in exercise capacity and Cit were found.


Evidence for the existence of stereoselective presynaptic beta 1-adrenoceptors on noradrenergic and dopaminergic neurons in the rat hypothalamus.
Using high performance liquid chromatography with an electrochemical detector, we examined antagonistic effects of l- and d-acebutolol and atenolol against isoproterenol-induced facilitation of impulse-evoked release of endogenous norepinephrine and dopamine in rat hypothalamic slices. Isoproterenol (10 nM) increased the release of the both catecholamines, and this facilitation was similarly antagonized by 100 nM l-acebutolol and 100 nM atenolol, but was not antagonized by 100 nM d-acebutolol. In conclusion, there exist stereoselective presynaptic beta 1-adrenoceptors on noradrenergic and dopaminergic neurons in the rat hypothalamus.


Comparative viabilities of rat intestinal epithelial cells prepared by mechanical, enzymatic and chelating methods.
Mechanical, enzymatic and chelating methods for isolating rat intestinal epithelial cells were compared to determine the best technique for obtaining high yields of viable cells. The mechanical techniques resulted in consistently poor viability compared with the other methods. There was excellent agreement between viability determinations made by trypan blue exclusion and by cytofluorochromasia. Incubation with EDTA, citrate and dithiothreitol, combined with mechanical extrusion of dissociated cells provided the highest yields of viable cells in these studies.


[Cryoconservation of 8-celled stage rabbit embryos in an automated "open system"].
Rabbit embryos were successfully frozen at the 8-cell stage, employing a rapid and a slow freezing-thawing program. They were then stored at -196 degrees C (liquid nitrogen) for 10 to 60 days. After thawing the embryos were examined for viability in vitro and in vivo. Using the slow program B 82.5% were morphologically intact, compared to only 69.3% after the rapid procedure (program A). 88.8% of the group first mentioned above, and 56.8% of the latter developed to the blastocyst stage after 4 to 5 days in culture. The implantation rate was 18.1%, when embryos were transferred to minus 18 hours asynchronous recipients, after thawing. With 40%, a significantly higher implantation rate could be achieved, when the embryos were transferred to minus 24 hours asynchronous foster mothers. 18 viable young were born 30 to 32 days after transfer.


CICIoT2023: A Real-Time Dataset and Benchmark for Large-Scale Attacks in IoT Environment.
Nowadays, the Internet of Things (IoT) concept plays a pivotal role in society and brings new capabilities to different industries. The number of IoT solutions in areas such as transportation and healthcare is increasing and new services are under development. In the last decade, society has experienced a drastic increase in IoT connections. In fact, IoT connections will increase in the next few years across different areas. Conversely, several challenges still need to be faced to enable efficient and secure operations (e.g., interoperability, security, and standards). Furthermore, although efforts have been made to produce datasets composed of attacks against IoT devices, several possible attacks are not considered. Most existing efforts do not consider an extensive network topology with real IoT devices. The main goal of this research is to propose a novel and extensive IoT attack dataset to foster the development of security analytics applications in real IoT operations. To accomplish this, 33 attacks are executed in an IoT topology composed of 105 devices. These attacks are classified into seven categories, namely DDoS, DoS, Recon, Web-based, brute force, spoofing, and Mirai. Finally, all attacks are executed by malicious IoT devices targeting other IoT devices. The dataset is available on the CIC Dataset website.


Analysis of the dose calculation accuracy for IMRT in lung: a 2D approach.
The purpose of this study was to compare the dosimetric accuracy of IMRT plans for targets in lung with the accuracy of standard uniform-intensity conformal radiotherapy for different dose calculation algorithms. Tests were performed utilizing a special phantom manufactured from cork and polystyrene in order to quantify the uncertainty of two commercial TPS for IMRT in the lung. Ionization and film measurements were performed at various measuring points/planes. Additionally, single-beam and uniform-intensity multiple-beam tests were performed, in order to investigate deviations due to other characteristics of IMRT. Helax-TMS V6.1(A) was tested for 6, 10 and 25 MV and BrainSCAN 5.2 for 6 MV photon beams, respectively. Pencil beam (PB) with simple inhomogeneity correction and 'collapsed cone' (CC) algorithms were applied for dose calculations. However, the latter was not incorporated during optimization hence only post-optimization recalculation was tested. Two-dimensional dose distributions were evaluated applying the gamma index concept. Conformal plans showed the same accuracy as IMRT plans. Ionization chamber measurements detected deviations of up to 5% when a PB algorithm was used for IMRT dose calculations. Significant improvement (deviations approximately 2%) was observed when IMRT plans were recalculated with the CC algorithm, especially for the highest nominal energy. All gamma evaluations confirmed substantial improvement with the CC algorithm in 2D. While PB dose distributions showed most discrepancies in lower (<50%) and high (>90%) dose regions, the CC dose distributions deviated mainly in the high dose gradient (20-80%) region. The advantages of IMRT (conformity, intra-target dose control) should be counterbalanced with possible calculation inaccuracies for targets in the lung. Until no superior dose calculation algorithms are involved in the iterative optimization process it should be used with great care. When only PB algorithm with simple inhomogeneity correction is used, lower energy photon beams should be utilized.


Structural characterization of a A-type linked trimeric anthocyanin derived pigment occurring in a young Port wine.
A new trimeric flavylium derivative pigment was detected for the first time in a young Port wine and structurally characterised using both API-LC ESI MS/MS and NMR techniques. Its structure comprises a malvidin-3-glucoside molecule (terminal unit) linked to two units of flavans (extension units) involving two A-type linkages (C4→C8 and C7-O-C2 ether linkages). The energies of the minimized geometries for all four conformations of this compound were calculated and comparing the structures of all isomers, it was verified that the binding mode (α or β) between the first and second anthocyanin unit does not have a key role in the compound stability. However, for isomers 1 and 2 where the interaction between the second and third anthocyanin units occurs by a "β" mode in opposition to the "α" mode verified in isomers 3 and 4, revealed a higher stability. The presence of these compounds in red wines is thought to result from their extraction from grape skin during fermentation, as the presence of these trimeric pigments was detected in a grape skin extract by API-LC ESI MS/MS spectrometry.


ATYPICAL MACULAR HOLES.
To study the etiology, clinical features, management options, and visual prognosis in various types of atypical macular holes (MHs). A review of the literature was performed, which focused on the etiopathogenesis of atypical or secondary MHs, their differentiating clinical features, management strategies, and varied clinical outcomes. Idiopathic or age-related, myopic, and traumatic MHs were excluded. Atypical or secondary MHs arise out of concurrent ocular pathologies (dystrophy, degeneration, or infections) and laser/surgery. The contributing factors may be similar to those responsible for idiopathic or typical MHs, i.e., tangential or anteroposterior vitreofoveal traction or cystoid degeneration. The management is either observation or treatment of the underlying cause. The prognosis depends on the background pathology, duration of disease, and baseline visual acuity governed by the size of MH and morphologic health of underlying RPE and photoreceptors. The closer the morphology of atypical MH is to that of an idiopathic MH, the better the surgical outcome is. With the advancements in retinal imaging, atypical MHs are now more frequently recognized. With increasing understanding of the underlying disease processes, and improvement in investigations and surgical treatment, management of atypical MHs may improve in the future.


NOD/LtSz-Rag1nullPfpnull mice: a new model system with increased levels of human peripheral leukocyte and hematopoietic stem-cell engraftment.
A critical need exists for effective small-animal models that accept engraftment of human hematopoietic progenitor cells and mature lymphocytes. The purpose of this study was to determine the phenotypic effects of perforin (Pfp) deficiency on nonobese diabetic (NOD)-Rag1null mice and to evaluate the ability of NOD/LtSz-Rag1nullPfpnull recipients to support engraftment with human hematolymphoid cells. A new genetic stock of NOD mice doubly homozygous for targeted mutations at the recombination activating gene (Rag)-1 and Pfp genes was developed. NOD/LtSz-Rag1nullPfpnull mice were studied for immunopathologic and hematologic abnormalities. The ability of these mice to support engraftment with human peripheral blood mononuclear cells (PBMC) and umbilical-cord blood hematopoietic progenitor cells was assessed. NOD/LtSz-Rag1nullPfpnull mice lacked mature B cells, T cells, natural killer (NK) cell cytotoxic activity and were devoid of serum immunoglobulin (Ig) throughout a 37-week lifespan. These mice supported heightened engraftment with human PBMC as compared with NOD/LtSz-Rag1null controls as evidenced by a 4- to 5-fold increase in percentages of human lymphocytes and a 7- to 13-fold increase in percentages of CD4+ T cells in the peripheral blood and spleen. Total numbers of human CD4+ T cells were increased approximately 20-fold in the spleens of NOD/LtSz-Rag1nullPfpnull mice. These mice also showed approximately 12-fold higher levels of engraftment with human umbilical-cord blood cells compared with NOD/LtSz-Rag1null mice. NOD/LtSz-Rag1nullPfpnull mice are devoid of mature B cell, T cell, and NK cell cytotoxic activity, engraft at high levels with human PBMC, and hematopoietic progenitor cells and provide a new NK cell-deficient model for human hematolymphoid cell engraftment.


[Generalised Form of Synovial Chondromatosis of the Knee Joint].
This study describes a diagnostic and therapeutic algorithm in a 53-year-old male patient who was diagnosed with a synovial chondromatosis of the knee joint extending to the popliteal fossa and soft tissues around the knee. Because of the presence of massive nodules, the patient was indicated for total synovectomy, with removal of pathologically changed cartilaginous tissue, performed by combined anterior and posterior approaches to the knee joint. Despite complete removal of the synovium and loose cartilage bodies and the patient's pain relief in the post-operative time, three years after the operation new problems appeared. Magnetic resonance imaging (MRI) confirmed a relapse of synovial chondromatosis and the patient was indicated for revision surgery of the knee joint. The results of physical examination and MRI scans, and intra-operative findings in the patient are reported. synovial chondromatosis, total synovectomy, direct anterior and posterior approaches to the knee joint.


A progressive radiorenographic (99mTc-DTPA) presentation of surgically induced renal vein hypertension in baboons.
A 99mTc-DTPA radiorenographic study, supported by excretory urography, blood chemistry and eventually histology was performed on 8 chacma baboons (Papio ursinus) after surgical induction of unilateral renal vein hypertension. Frequent sequential repeat renograms during 6 post-operative months detected fluctuating renal behaviour but did not reveal typical course of the disease. Neither did any one of the radiorenographic parameters (TP, T1/2, FF, relative clearances and perfusion) prove to be more sensitive than the others in detecting abnormalities. Initial microscopic or macroscopic haematuria and proteinuria in general eventually cleared up. Light microscopy demonstrated only mild abnormalities. The effects of venous occlusion of the kidney and the prognosis seemed to be variable as was reflected by radiorenography.


Larger, unfiltered datasets are more effective at resolving phylogenetic conflict: Introns, exons, and UCEs resolve ambiguities in Golden-backed frogs (Anura: Ranidae; genus Hylarana).
Using FrogCap, a recently-developed sequence-capture protocol, we obtained >12,000 highly informative exons, introns, and ultraconserved elements (UCEs), which we used to illustrate variation in evolutionary histories of these classes of markers, and to resolve long-standing systematic problems in Southeast Asian Golden-backed frogs of the genus-complex Hylarana. We also performed a comprehensive suite of analyses to assess the relative performance of different genetic markers, data filtering strategies, tree inference methods, and different measures of branch support. To reduce gene tree estimation error, we filtered the data using different thresholds of taxon completeness (missing data) and parsimony informative sites (PIS). We then estimated species trees using concatenated datasets and Maximum Likelihood (IQ-TREE) in addition to summary (ASTRAL-III), distance-based (ASTRID), and site-based (SVDQuartets) multispecies coalescent methods. Topological congruence and branch support were examined using traditional bootstrap, local posterior probabilities, gene concordance factors, quartet frequencies, and quartet scores. Our results did not yield a single concordant topology. Instead, introns, exons, and UCEs clearly possessed different phylogenetic signals, resulting in conflicting, yet strongly-supported phylogenetic estimates. However, a combined analysis comprising the most informative introns, exons, and UCEs converged on a similar topology across all analyses, with the exception of SVDQuartets. Bootstrap values were consistently high despite high levels of incongruence and high proportions of gene trees supporting conflicting topologies. Although low bootstrap values did indicate low heuristic support, high bootstrap support did not necessarily reflect congruence or support for the correct topology. This study reiterates findings of some previous studies, which demonstrated that traditional bootstrap values can produce positively misleading measures of support in large phylogenomic datasets. We also showed a remarkably strong positive relationship between branch length and topological congruence across all datasets, implying that very short internodes remain a challenge to resolve, even with orders of magnitude more data than ever before. Overall, our results demonstrate that more data from unfiltered or combined datasets produced superior results. Although data filtering reduced gene tree incongruence, decreased amounts of data also biased phylogenetic estimation. A point of diminishing returns was evident, at which higher congruence (from more stringent filtering) at the expense of amount of data led to topological error as assessed by comparison to more complete datasets across different genomic markers. Additionally, we showed that applying a parameter-rich model to a partitioned analysis of concatenated data produces better results compared to unpartitioned, or even partitioned analysis using model selection. Despite some lingering uncertainties, a combined analysis of our genomic data and sequences supplemented from GenBank (on the basis of a few gene regions) revealed highly supported novel systematic arrangements. Based on these new findings, we transfer Amnirana nicobariensis into the genus Indosylvirana; and I. milleti and Hylarana celebensis to the genus Papurana. We also provisionally place H. attigua in the genus Papurana pending verification from positively identified (voucher substantiated) samples.


Challenges of Huntington's disease and quest for therapeutic biomarkers.
Huntington's disease (HD) is the most common inherited neurodegenerative disorder among polyglutamine (polyQ) diseases caused by cytosine-adenine-guanine repeat expansion in exon 1 of the huntingtin gene whose translation results in polyQ stretch in the N-terminus of the huntingtin protein (HD protein). This mutation significantly affects huntingtin conformation, proteolysis, PTMs, as well as its ability to bind interacting proteins. As a consequence, a variety of cellular mechanisms such as transcription, mitochondrial energy metabolism, axonal transport, neuronal vulnerability to oxidative stress, neurotransmission, and immune response are altered and involved in the pathogenesis of HD. Promising candidate molecular biomarkers of HD have emerged from proteomic studies. Recent analyses focused on HD protein itself, its PTM, and interacting proteins, which are of great importance for disease course. Furthermore, brain, body fluids, and immune system are intensively studied in order to search for additional proteins with a view to their use as a biomarker(s) or set of biomarkers in clinical trials in HD translational research.


EK (DLEK, DSEK, DMEK): New Frontier in Cornea Surgery.
Endothelial keratoplasty (EK) has revolutionized treatment of corneal endothelial dysfunction. Compared with penetrating keratoplasty (PK), EK provides faster and more reliable visual rehabilitation while maintaining the eye's structural integrity. The number of EK procedures is growing annually and surpassed PK in the United States in 2012. The most widely used iteration, Descemet stripping endothelial keratoplasty (DSEK), implants healthy donor endothelium, Descemet membrane, and posterior stroma. Descemet membrane endothelial keratoplasty (DMEK) eliminates the donor stromal layer. Although more surgically challenging than DSEK, DMEK provides even faster visual rehabilitation and reduced risk of immunologic rejection, so its use is growing. Potential future alternatives to EK that could help address the unmet demand for donor corneas include removing central guttae and regenerating a central endothelial cell layer from healthy peripheral cells in patients with Fuchs' dystrophy or injecting cultured human corneal endothelial cells to rehabilitate eyes without residual healthy endothelium.


Optimization of human mesenchymal stem cell isolation from synovial membrane: Implications for subsequent tissue engineering effectiveness.
Synovium-derived mesenchymal stem cells (SDMSCs) are one of the most suitable sources for cartilage repair because of their chondrogenic and proliferative capacity. However, the isolation methods for SDMSCs have not been extensively characterized. Thus, our aim in this study was to optimize the processes of enzymatic isolation followed by culture expansion in order to increase the number of SDMSCs obtained from the original tissue. Human synovium obtained from 18 donors (1.5 g/donor) was divided into three aliquots. The samples were minced and subjected to collagenase digestion, followed by different procedures: Group 1, Tissue fragments were removed by filtering followed by removing floating tissue; Group 2, No filtering. Only floating fragments were removed; Group 3, No fragments were removed. Subsequently, each aliquot was sub-divided into two density subgroups with half. In Group 1, the cell-containing media was plated either at high (5000 cells/cm2) or low density (1000 cells/cm2). In Groups 2 and 3, the media containing cells and tissue was plated onto the same number of culture dishes as used in Group 1, either at high or low density. At every passage, the cells plated at high density were consistently re-plated at high and those plated at low density were likewise. The expanded cell yields at day 21 following cell isolation were calculated. These cell populations were then evaluated for their osteogenic, adipogenic, and chondrogenic differentiation capabilities. The final cell yields per 0.25 g tissue in Group 1 were similar at high and low density, while those in Groups 2 and 3 exhibited higher when cultured at low density. The cell yields at low density were 0.7 ± 1.2 × 107 in Group 1, 5.7 ± 1.1 × 107 in Group 2, 4.3 ± 1.2 × 107 in Group 3 (Group 1 vs Groups 2 and 3, p < 0.05). In addition, the cells obtained in each low density subgroup exhibited equivalent osteogenic, adipogenic, and chondrogenic differentiation. Thus, it was evident that filtering leads to a loss of cells and does not affect the differentiation capacities. In conclusion, exclusion of a filtering procedure could contribute to obtain higher number of SDMSCs from synovial membrane without losing differentiation capacities.


Late-onset Sepsis in Extremely Premature Infants: 2000-2011.
Late-onset sepsis (LOS) is an important cause of death and neurodevelopmental impairment in premature infants. The purpose of this study was to assess overall incidence of LOS, distribution of LOS-causative organisms and center variation in incidence of LOS for extremely premature infants over time. In a retrospective analysis of infants 401-1000 g birth weight and 22-28 6/7 weeks of gestational age born at 12 National Institute of Child Health and Human Development Neonatal Research Network centers in the years 2000-2005 (era 1) or 2006-2011 (era 2) who survived >72 hours, we compared the incidence of LOS and pathogen distribution in the 2 eras using the χ test. We also examined the effect of birth year on the incidence of LOS using multivariable regression to adjust for nonmodifiable risk factors and for center. To assess whether the incidence of LOS was different among centers in era 2, we used a multivariable regression model to adjust for nonmodifiable risk factors. Ten-thousand one-hundred thirty-one infants were studied. LOS occurred in 2083 of 5031 (41%) infants in era 1 and 1728 of 5100 (34%) infants in era 2 (P < 0.001). Birth year was a significant predictor of LOS on adjusted analysis, with birth years 2000-2009 having a significantly higher odds of LOS than the reference year 2011. Pathogens did not differ, with the exception of decreased fungal infection (P < 0.001). In era 2, 9 centers had significantly higher odds of LOS compared with the center with the lowest incidence. The incidence of LOS decreased over time. Further investigation is warranted to determine which interventions have the greatest impact on infection rates.


Evaluation of the efficacy and safety of in vitro, adenovirus-mediated transfer of the human cystic fibrosis transmembrane conductance regulator cDNA.
Cystic fibrosis (CF) is a common, fatal recessive disease caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene manifested by abnormalities in the regulation of chloride ion (Cl-) secretion across the apical membrane of epithelial cells throughout the body. Adenovirus-mediated delivery of the normal CFTR cDNA and correction of the CF epithelial cell Cl- secretory phenotype suggests the feasibility of gene therapy for CF lung disease. Few studies, however, have focused on the evaluation of the safety of the adenovirus-mediated gene transfer approach. This study presents in vitro data on the efficacy and safety of adenovirus-mediated transfer of the human CFTR cDNA using Av1Cf2. Av1Cf2-mediated transfer of the human CFTR cDNA complemented the abnormal cAMP-regulated Cl- permeability of cells with the CF epithelial phenotype. Av1 vectors did not replicate infectious virus in HeLa cells infected in vitro, although trace vector DNA synthesis was observed at very high multiplicity of infection. Expression of the adenoviral late gene for the hexon capsid protein was observed at trace levels in Av1 vector-infected HeLa cells, but not in freshly isolated human bronchial epithelial cells, consistent with the pattern of DNA synthesis observed in these different target cells. Although, these observations support the efficacy and safety of use of Av1Cf2 for treatment of the fatal pulmonary component of CF.


[Comparative studies on identification of maize secreted proteome by different analyzing techniques].
Maize roots release a battery of proteins into the rhizosphere during growth and development. In the present study, mucilage secreted by primary roots of 3-4 d old maize seedlings was collected under axenic conditions, and the constitutively secreted proteins were analyzed by different methods, including two-dimensional electrophoresis--MADLI-TOF, LC-MS, and Shotgun techniques. The results showed that 200 protein spots could be observed on two-dimensional electrophoresis gel stained by silver nitrate. However, due to the low protein content for each protein spot, there was not reliable result after identification by MADLI-TOF. On the other hand, 152 and 2848 proteins were identified by LC/MS and Shotgun technique, respectively. All 152 proteins identified by LC/MS method could be found in the proteins identified by Shotgun method, and thus the results obtained by LC/MS and Shotgun method could be authenticated mutually. The results demonstrated that the Shotgun method is more sensitive than the other methods for protein identification, especially for the proteins with lower contents and complex interfering substances, such as the plant root released proteins.


Fibroblast growth factors signaling in bone metastasis.
Many solid tumors metastasize to bone, but only prostate cancer has bone as a single, dominant metastatic site. Recently, the FGF axis has been implicated in cancer progression in some tumors and mounting evidence indicate that it mediates prostate cancer bone metastases. The FGF axis has an important role in bone biology and mediates cell-to-cell communication. Therefore, we discuss here basic concepts of bone biology, FGF signaling axis, and FGF axis function in adult bone, to integrate these concepts in our current understanding of the role of FGF axis in bone metastases.


Human resident CD34+ stromal cells/telocytes have progenitor capacity and are a source of αSMA+ cells during repair.
We studied the progenitor capacity of human resident CD34+ stromal cells/telocytes (SC/TCs) in the enteric wall affected by inflammatory/repair processes (appendicitis, diverticulitis of large bowel and Crohn's disease of the terminal ileum) at different stages of evolution (inflammatory, proliferative and remodelling). In these conditions, CD34+ SC/TCs are activated, showing changes, which include the following overlapping events: 1) separation from adjacent structures (e.g., from vascular walls) and location in oedematous spaces, 2) morphological modifications (in cell shape and size) with presence of transitional cell forms between quiescent and activated CD34+ SC/TCs, 3) rapid proliferation and 4) loss of CD34 expression and gain of αSMA expression. These events mainly occur in the inflammatory and proliferative stages. During the loss of CD34 expression, the following findings are observed: a) irregular cell labelling intensity for anti-CD34, b) co-localization of CD34 and actin, c) concurrent irregular labelling intensity for αSMA and d) αSMA expression in all stromal cells, with total loss of CD34 expression. While CD34 expression was conserved, a high proliferative capacity (Ki-67 expression) was observed and vice versa. In the segments of the ileum affected by Crohn's disease, the stromal cells around fissures were αSMA+ and, in the transitional zones with normal enteric wall, activated CD34+ SC/TCs were observed. In conclusion, human resident CD34+ SC/TCs in the enteric wall have progenitor capacity and are activated with or without differentiation into αSMA+ stromal cells during inflammatory/repair processes.


Recurrent Mesenteric Ischemia from Celiomesenteric Trunk Stenosis.
A common origin of the celiac trunk and superior mesenteric artery (SMA), also termed as celiomesenteric trunk (CMT), is a rare occurrence. We report a rare case of symptomatic CMT stenosis requiring multiple interventions. The patient underwent an initial superior mesenteric artery bypass graft but required a subsequent endoluminal intervention. We present this case for the rarity of CMT stenosis and its potential to cause recurrent mesenteric ischemia. The treatment outcome in this patient suggests that revascularization of the SMA alone can result in adequate perfusion of the entire mesenteric bed and resolve symptoms of mesenteric ischemia including weight loss and food avoidance.


Gastroesophageal Reflux Disease and Pulmonary Diseases Associated with Aspergillosis: Is There a Connection?
We studied the relationship between GORD and allergic bronchopulmonary aspergillosis (ABPA), chronic pulmonary aspergillosis (CPA), or Aspergillus bronchitis. Gastroesophageal reflux disease (GORD) is well known to initiate or exacerbate pulmonary inflammatory conditions, inducing bronchial asthma and chronic obstructive lung disease. We reviewed four patients referred with elevated Aspergillus serology markers and marked pulmonary symptoms for ABPA, CPA, and Aspergillus bronchitis, and discussed the underlying pathophysiological relationship with GORD. Data were collected retrospectively from medical records included age, gender, predisposing factors for ABPA, chronic pulmonary aspergillosis, or Aspergillus bronchitis; presence of nocturnal reflux, nausea, epigastric pain, Medical Research Council dyspnea scale score, pH manometry data, endoscopic results (ulcers, Barrett's esophagus), treatment of GORD [proton-pump inhibitors (PPIs), surgical operation]; history of smoking, alcohol consumption; concomitant COPD; serological markers (anti-Aspergillus IgG, anti-Aspergillus IgE), and antifungal treatment. Four patients with GORD were studied; following PPIs administration two achieved clinical improvement. One had ABPA, one CPA, and two had Aspergillus bronchitis; median age was 57 years [range 39-71]; males-to-females ratio was 1:3. Serological markers for aspergillosis were: median total IgE antibodies 573 KIU/L [range 13.1-850], median Aspergillus IgE-specific antibodies 1.2 kAU/L [range <0.4-24.7], and median Aspergillus IgG titers 71 mg/L [range 20-119]. Aspergillus fumigatus grew in one sputum sample. Clinicians caring for patients with gastroesophageal reflux disease presenting with elevated Aspergillus IgG or IgE antibodies should maintain a high index of suspicion for this association and proceed to appropriate evaluations, including laryngoscopy and endoscopy, initiating specific PPI-directed therapy when indicated.


Bacterial expression of two human aryl sulfotransferases.
The effect of replacing a single codon in the N-terminal of human aryl sulfotransferase (HAST) 1 and 3 with one that is more commonly found in E. coli genes was assessed. The pKK233-2 E. coli expression vector was employed and the polymerase chain reaction (PCR) was used to introduce the 5' nucleotide substitution, at the same time maintaining the fidelity of the amino acid sequence. The data indicates that this change had a minimal effect on protein production, subcellular localization or, in the case of HAST3, catalytic activity. In general, the pKK233-2 E. coli vector has been less than optimal for expressing human sulfotransferase cDNAs.


Clostridium punense sp. nov., an obligate anaerobe isolated from healthy human faeces.
An obligately anaerobic, rod-shaped (0.5-1.0 × 2.0-10.0 μm), Gram-stain-positive bacterium, occurring mainly singly or in pairs, and designated BLPYG-8T, was isolated from faeces of a healthy human volunteer aged 56 years. Cells were non-motile. Oval, terminal spores were formed that swell the cells. The strain was affiliated with the genus Clostridium sensu stricto (Clostridium rRNA cluster I) as revealed by 16S rRNA gene sequence analysis. Strain BLPYG-8T showed 97.3 to 97.4 % 16S rRNA gene sequence similarity with Clostridium sulfidigenes DSM 18982T, Clostridium subterminale DSM 6970T and Clostridium thiosulfatireducens DSM 13105T. DNA-DNA hybridization and phenotypic analysis showed that the strain was distinct from its closest relatives, C. sulfidigenes DSM 18982T, C. subterminale DSM 6970T, C. thiosulfatireducens DSM 13105T with 54.2, 53.9 and 53.3 % DNA-DNA relatedness, respectively. Strain BLPYG-8T grew in PYG broth at temperatures between 20 and 40 °C (optimum 37 °C). The strain utilized a range of amino acids as well as carbohydrates as a source of carbon and energy. Glucose fermentation resulted in the formation of volatile fatty acids mainly acetic acid, n-butyric acid and organic acids such as succinic and lactic acid. The DNA G+C content of strain BLPYG-8T was 44.1 mol%. The major fatty acids (>10 %) were C14 : 0, iso-C15 : 0, C16 : 1ω7c and C16 : 0. Phylogenetic analysis and specific phenotypic characteristics and/or DNA G+C content differentiated the strain from its closest relatives. On the basis of these data, strain BLPYG-8T represents a novel species of the genus Clostridium, for which the name Clostridium punense sp. nov. is proposed. The type strain is BLPYG-8T ( = DSM 28650T = CCUG 64195T = MCC 2737T).


Correlation between telomerase activity and DNA ploidy in gastric cancer.
Telomerase has been reported to be activated in most immortal cells and human cancers. In the present study, we assessed the correlation between telomerase activity and cellular DNA ploidy level in gastric cancer. Telomerase activity was determined semiquantitatively using the telomeric repeat amplification protocol assay, a polymerase-chain-reaction-based assay, in surgical specimens of primary tumors obtained from 36 patients with gastric cancer. No correlation was observed between telomerase activity and the proliferating cell nuclear antigen labeling index. In contrast, a positive linear correlation was observed between telomerase activity and the DNA index (r = 0.59; p < 0.01). Tumor cells with aneuploid patterns showed higher telomerase activity than those with diploid patterns (27.6+/-5.8 vs. 5.8+/-1.1%; p < 0.01). Telomerase activity of tumors with liver metastases was significantly higher than activity of those without metastases (34.5+/-16.6 vs. 11.8+/-2.4; p < 0.05). There was a trend toward a lower survival rate in 9 patients with a telomerase activity of 20% or higher compared to 27 patients with telomerase activity lower than 20%. These results suggest that the telomerase activity of gastric cancer tissue may reflect the malignant potential of the tumor.


Uterine smooth-muscle tumors with unusual growth patterns: imaging with pathologic correlation.
This essay illustrates the salient features of variant smooth-muscle tumors on multiple imaging techniques with correlative pathology. We describe how recognition of these features allows the radiologist to distinguish a uterine leiomyoma variant from the classic fibroid or a leiomyosarcoma. Finally, we highlight the role of the radiologist in triaging these patients to surgical versus medical management and in surgical planning. Parasitic leiomyoma, intravenous leiomyomatosis, disseminated peritoneal leiomyomatosis, and benign metastasizing leiomyoma show key features on multiple imaging techniques that correlate with pathology findings. In the appropriate clinical setting, the radiologist should include these unusual lesions in the broader differential diagnosis of smooth-muscle tumors and, in certain cases, aid in surgical planning.


A cross-sectional analysis of the association between diabetes self-care practices and blood glucose regulation among Egyptian diabetic patients.
Diabetes Mellitus (DM) is a main public health issue worldwide, with Egypt among the world's top countries with diabetic patients. Data on the degree of self-care behaviors of Egyptian diabetic patients is not fully reported. Therefore, the study aimed to assess the adequacy of diabetes self-management (DSM) habits among diabetic patients in Egypt and to identify associated socio-demographic factors that could negatively impact the patients' glycemic state, using a structured questionnaire. This study was a cross-sectional observational study. The diabetes self-management questionnaire (DSMQ) was used to assess diabetes self-care activities, which is a 16-item questionnaire. The statistical analysis was conducted on SPSS software, and the data were significant with a p-value <0.05. A total of 2,164 participants were included. The majority of the patients (83.8%) had type 2 DM. The mean glycated hemoglobin (HbA1c) of the study group was 8.44±1.99. Significant differences were found between different glycemic control groups both in terms of the DSMQ sum scores and the subscale scores, with the exception of physical activity. Moreover, there was a significant weak inverse correlation between the total DSMQ sum score and HbA1c ≥9.0% (ρ=-0.116, p<0.01). The overall internal consistency (Cronbach's alpha) was acceptable (0.756). The study indicated that patients with controlled blood glucose had significantly higher 'glucose management', and 'total DSMQ sum' scores compared to uncontrolled diabetic patients. All significant correlations between assessed parameters and DSMQ were weak.


Intraoperative MR systems. High-field approaches.
Since the introduction of MR-guided neurosurgery, the field has evolved considerably. As it has been convincingly demonstrated, MR-guided therapy seems to be feasible and safe at all clinical field strengths up to 1.5 T. This article discusses the technology of MR-guided therapy at 1.5 T, and in particular, the multiple approaches that have been advanced to date.


Upregulation of jasmonate biosynthesis and jasmonate-responsive genes in rice leaves in response to a bacterial pathogen mimic.
Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight of rice, secretes several cell wall degrading enzymes including cellulase (ClsA) and lipase/esterase (LipA). Prior treatment of rice leaves with purified cell wall degrading enzymes such as LipA can confer enhanced resistance against subsequent X. oryzae pv. oryzae infection. To understand LipA-induced rice defense responses, microarray analysis was performed 12 h after enzyme treatment of rice leaves. This reveals that 867 (720 upregulated and 147 downregulated) genes are differentially regulated (≥2-fold). A number of genes involved in defense, stress, signal transduction, and catabolic processes were upregulated while a number of genes involved in photosynthesis and anabolic processes were downregulated. The microarray data also suggested upregulation of jasmonic acid (JA) biosynthetic and JA-responsive genes. Estimation of various phytohormones in LipA-treated rice leaves demonstrated a significant increase in the level of JA-Ile (a known active form of JA) while the levels of other phytohormones were not changed significantly with respect to buffer-treated control. This suggests a role for JA-Ile in cell wall damage induced innate immunity. Furthermore, a comparative analysis of ClsA- and LipA-induced rice genes has identified key rice functions that might be involved in elaboration of damage-associated molecular pattern (DAMP)-induced innate immunity.


Effect of Class II functional treatment on facial attractiveness, as perceived by professionals and laypeople.
The literature offers different perspectives for and against two-phase treatment of skeletal Class II malocclusion. Facial attractiveness is an important aspect to take into account, given that children with skeletal Class II are often bullied by their peers and have low self-esteem and a lower social perception. The aim of the present study was to evaluate the aesthetic perception of facial profiles by a large number of observers, before and after treatment with a functional appliance, compared to untreated controls. The pre- and post-treatment cephalograms of 20 Class II subjects treated with Sander's bite-jumping appliance and 20 untreated historical controls were collected and transformed into black and white silhouettes depicting only the lower third of the face. An online questionnaire comprising the silhouettes of the two groups, three "calibration" profiles and an "ideal" profile was submitted to dentists, orthodontists, undergraduates and laypeople, asking them to rate the profile's attractiveness using a Visual Analogue Scale (VAS). The effect of treatment, and observers' age, expertise and gender were analysed. The calibration images and the ideal profiles were used to evaluate the coherence of each observer's judgement. The protocol was approved by the local Ethics Committee. Nine-hundred and ten questionnaires were collected. Treated subjects showed a larger improvement of facial attractiveness compared to controls. A significant effect of gender on the observer's ratings was observed. Some observers showed incoherent judgement, which had a significant effect on the regression model. In conclusion, early treatment with functional appliances seems to improve patients' facial aesthetics. This improvement is perceived equally by dental professionals and laypeople.


Teachers, not parents, are able to predict time processing skills in preschoolers.
Time processing difficulties are associated with developmental disorders. Questionnaires for assessing children's sense of time are available from primary school, but we lack valid proxy-report tools for younger children, who are not able to complete self-reports. This study aimed to assess the criterion validity of a questionnaire investigating preschoolers' sense of time from the points of view of their parents and teachers. One hundred seventy preschoolers were included in the sample. Their parents and teachers completed the Sense of Time Questionnaire, and the children were administered time reproduction and time discrimination tasks, both concurrently and 7 months later. The assessment of preschoolers' sense of time reported by teachers, but not by parents, predicted the children's time processing skills both concurrently and longitudinally. The teacher version of the Sense of Time Questionnaire constitutes a valid instrument for assessing and predicting preschoolers' time processing skills and can be used for clinical and research purposes STATEMENT OF CONTRIBUTION: What is already known on this subject? Time processing difficulties are associated with developmental disorders such as ADHD and dyscalculia. Early assessment of time processing skills is important for clinical (e.g., screening) and research purposes. We do not have valid questionnaires for assessing sense of time in young children. What the present study adds? The sense of time ability reported by teachers predicts preschoolers' time processing skills. The sense of time ability reported by parents does not predicts preschoolers' time processing skills. The Sense of Time Questionnaire is a valid instrument for investigating time processing skills of 4-6 aged children.


A single-centre audit of junior doctors' diagnostic activity in medical admissions.
Practising doctors are distinguished from other healthcare staff by their role in making a clinical diagnosis. Huge changes in training and working practice in the past 15 years may have left many junior doctors ill equipped or preferring not to synthesise information to conclude a clerking with a diagnosis or differential diagnosis. This report details a retrospective study of acute medical admissions (AMU) first seen in the emergency department (A&E) and the diagnostic activity of junior doctors in both settings. The documented diagnostic conclusion and reported differential diagnosis was compared with that of the relevant admission consultant. The aim was to see if doctors completed their clinical assessment by establishing a 'reasonable differential diagnosis' or simply used the presenting complaint (a symptom) of the patient as the 'diagnosis'. One hundred patients' records (66 male and 34 female of mean age 57.4 years [20-94 years]) were studied. The majority of cases came from the cardiac and neurology domains of diagnosis. A total of 53% of cases seen in A&E by clinicians were given a clinical 'symptom' as a final diagnosis or differential diagnosis. A further 18% referred to the wrong clinical domain in their assessment, and 22% of cases were concordant with the eventual consultant diagnosis. In the AMU 20% of cases were given a symptom as a final diagnosis and 11% the wrong domain; 45% of diagnoses were concordant. The grade of the doctor, from foundation year (FY1) to specialty registrar (ST3), led to an expected improvement in the assessment and documentation of a possible diagnosis rather than a symptom for both A&E and AMU settings. In summary, junior doctors did not routinely document a clinical diagnosis or differential diagnosis at the conclusion of their clerking, regardless of experience. The reasons for this deferred activity are considered.


Targeted Early Detection of Citrus Huanglongbing Causal Agent 'Candidatus Liberibacter asiaticus' Before Symptom Expression.
Citrus Huanglongbing (HLB) is the most severe disease of citrus plants caused by 'Candidatus Liberibacter asiaticus' and transmitted by the insect vector Asian citrus psyllid (ACP). No effective curative measure is available against HLB. For citrus production areas without HLB or with low HLB disease incidence, removal of 'Ca. L. asiaticus' inoculum is critical to prevent HLB spread. Such a strategy requires robust early diagnosis of HLB for inoculum removal to prevent ACP acquisition and transmission of 'Ca. L. asiaticus'. However, early diagnosis of HLB is challenging, because the citrus trees remain asymptomatic for several months to years after 'Ca. L. asiaticus' transmission by ACP. In this study, we report a new method for targeted early detection of 'Ca. L. asiaticus' in cultivar Valencia sweet orange (Citrus sinensis) before HLB symptom expression. We take advantage of the fact that 'Ca. L. asiaticus' remains around the ACP feeding site immediately after transmission into the young flush and before flush maturation. ACPs secrete salivary sheaths at their feeding sites, which can be visualized using Coomassie brilliant blue staining owing to the presence of salivary sheaths secreted by ACP. Epifluorescence and confocal microscopy indicate the presence of salivary sheaths beneath the blue spots on ACP-fed leaves. Quantitative real-time polymerase chain reaction (PCR) and conventional PCR assays are able to detect 'Ca. L. asiaticus' in the ACP feeding surrounding areas as early as 2 to 20 days after ACP feeding. This finding lays a foundation to develop much-needed tools for early diagnosis of HLB before symptom expression, thus assisting 'Ca. L. asiaticus' inoculum removal and preventing HLB from spreading.


Polymeric inserts differing in their chemical composition as substrates for dendritic cell cultivation.
Dendritic cells (DC) contribute to immunity by presenting antigens to T cells and shape the immune response by the secretion of cytokines. Due to their immune stimulatory potential DC-based therapies are promising approaches to overcome tolerance e.g. against tumors. In order to enforce the immunogenicity of DCs, they have to be matured and activated in vitro, which requires an appropriate cell culture substrate, supporting their survival expansion and activation.Since most cell culture devices are not optimized for DC growth, it is hypothesized that polymers with certain physicochemical properties can positively influence the DC cultures. With the aim to evaluate the effects that polymers with different chemical compositions have on the survival, the activation status, and the cytokine/chemokine secretion profile of DC, their interaction with polystyrene (PS), polycarbonate (PC), poly(ether imide) (PEI), and poly(styrene-co-acrylonitrile) (PSAN)-based cell culture inserts was investigated. By using this insert system, which fits exactly into 24 well cell culture plates, effects induced from the culture dish material can be excluded. The viability of untreated DC after incubation with the different inserts was not influenced by the different inserts, whereas LPS-activated DC showed an increased survival after cultivation on PC, PS, and PSAN compared to tissue culture polystyrene (TCP). The activation status of DC estimated by the expression of CD40, CD80, CD83, CD86 and HLA-DR expression was not altered by the different inserts in untreated DC but slightly reduced when LPS-activated DC were cultivated on PC, PS, PSAN, and PEI compared to TCP. For each polymeric cell culture insert a distinct cytokine profile could be observed.Since inserts with different chemical compositions of the inserts did not substantially alter the behavior of DC all insert systems could be considered as alternative substrate. The observed increased survival on some polymers, which showed in contrast to TCP a hydrophobic surface, could be beneficial for certain applications such as T cell expansion and activation.


Differential responses to steroid hormones in fibroblasts from the vocal fold, trachea, and esophagus.
There is accumulating evidence that fibroblasts are target cells for steroids such as sex hormones and corticoids. The characteristics of fibroblasts vary among tissues and organs. Our aim in this study is to examine differences in responses to steroid hormones among fibroblasts from different cervicothoracic regions. We compared the actions of steroid hormones on cultured fibroblasts from the vocal folds, which are considered to be the primary target of steroid hormones, and the trachea and esophagus in adult male rats. Expression of steroid hormone receptors (androgen receptor, estrogen receptor α, and glucocorticoid receptor) was identified by immunofluorescence histochemistry. Androgen receptor was much more frequently expressed in fibroblasts from the vocal fold than in those from the trachea and esophagus. Cell proliferation analysis showed that administration of testosterone, estradiol, or corticosterone suppressed growth of all 3 types of fibroblasts. However, mRNA expression for extracellular matrix-associated genes, including procollagen I and III and elastin, and hyaluronic acid synthase I was elevated only by addition of testosterone to fibroblasts from the vocal fold. These results indicate that each steroid hormone exerts region-specific effects on cervicothoracic fibroblasts with different properties through binding to specific receptors.


Delignified and Densified Cellulose Bulk Materials with Excellent Tensile Properties for Sustainable Engineering.
Today's materials research aims at excellent mechanical performance in combination with advanced functionality. In this regard, great progress has been made in tailoring the materials by assembly processes in bottom-up approaches. In the field of wood-derived materials, nanocellulose research has gained increasing attention, and materials with advanced properties were developed. However, there are still unresolved issues concerning upscaling for large-scale applications. Alternatively, the sophisticated hierarchical scaffold of wood can be utilized in a top-down approach to upscale functionalization, and one can profit at the same time from its renewable nature, CO2 storing capacity, light weight, and good mechanical performance. Nevertheless, for bulk wood materials, a wider multipurpose industrial use is so far impeded by concerns regarding durability, natural heterogeneity as well as limitations in terms of functionalization, processing, and shaping. Here, we present a novel cellulose bulk material concept based on delignification and densification of wood resulting in a high-performance material. A delignification process using hydrogen peroxide and acetic acid was optimized to delignify the entire bulk wooden blocks and to retain the highly beneficial structural directionality of wood. In a subsequent step, these cellulosic blocks were densified in a process combining compression and lateral shear to gain a very compact cellulosic material with entangled fibers while retaining unidirectional fiber orientation. The cellulose bulk materials obtained by different densification protocols were structurally, chemically, and mechanically characterized revealing superior tensile properties compared to native wood. Furthermore, after delignification, the cellulose bulk material can be easily formed into different shapes, and the delignification facilitates functionalization of the bioscaffold.


Nucleus-encoded plastid sigma factor SIG3 transcribes specifically the psbN gene in plastids.
We have investigated the function of one of the six plastid sigma-like transcription factors, sigma 3 (SIG3), by analysing two different Arabidopsis T-DNA insertion lines having disrupted SIG3 genes. Hybridization of wild-type and sig3 plant RNA to a plastid specific microarray revealed a strong reduction of the plastid psbN mRNA. The microarray result has been confirmed by northern blot analysis. The SIG3-specific promoter region has been localized on the DNA by primer extension and mRNA capping experiments. Results suggest tight regulation of psbN gene expression by a SIG3-PEP holoenzyme. The psbN gene is localized on the opposite strand of the psbB operon, between the psbT and psbH genes, and the SIG3-dependent psbN transcription produces antisense RNA to the psbT-psbH intergenic region. We show that this antisense RNA is not limited to the intergenic region, i.e. it does not terminate at the end of the psbN gene but extends as antisense transcript to cover the whole psbT coding region. Thus, by specific transcription initiation at the psbN gene promoter, SIG3-PEP holoenzyme could also influence the expression of the psbB operon by producing psbT antisense RNA.


Three-dimensional in vivo imaging of tumors expressing red fluorescent proteins.
3D imaging of genetically-engineered fluorescent tumors enables quantitative monitoring of tumor growth/regression, metastatic processes, including during anticancer therapy in real-time.Fluorescent tumor models for 3D imaging require stable expression of genetically encoded fluorescent proteins and maintenance of the properties of tumor cell line including growth rate, morphology, and immunophenotype.In this chapter, the protocol for 3D imaging of tumors expressing red fluorescent protein are described in detail.


Refraction of water waves by periodic cylinder arrays.
We show that in the long wavelength limit, water waves propagate through an array of bottom-mounted vertical cylinders as if the water has an effective depth and effective gravitational constant that depends on the filling ratio of the cylinders, leading to refraction phenomena that can be described by analytic formulas. The results are obtained with rigorous homogenization techniques, as well as the multiple scattering formalism that gives full dispersion relationships. This phenomenon provides a mechanism to control the flow of ocean wave energy, as exemplified by a water-wave focusing lens.


Long-term outcomes of blebs repaired with scleral patch graft and conjunctival advancement in late-onset leak post-trabeculectomy.
To report long-term outcomes in eyes that developed late-onset bleb leak post trabeculectomy, with or without hypotony and/or maculopathy, due to a scleral melt/fistula and who required a scleral patch graft and conjunctival advancement for repair. Retrospective, non-comparative, interventional case series over a decade (2010-2019), presenting with late bleb leak post-filtration-surgery. All cases required a scleral patch graft and conjunctival advancement for management via a standard technique, performed by an experienced glaucoma surgeon. A total of 18 eyes were included. Mean age was 51.5 ± 10.2 years (95% CI [46.4-56.7]) and were followed up after repair for 52.4 ± 26.9 months, 95%CI [39.1-65.8]. 66.7% eyes (n = 12) had IOP ≦6 mmHg and also had hypotony maculopathy. None of the eyes presented with blebitis. 44.5% (n = 8) eyes underwent phacoemulsification as significant cataract was present. LogMAR best-corrected visual acuity (BCVA) was 0.8 ± 0.7 (95% CI [0.4-1.1]) prior to intervention and improved to 0.4 ± 0.6 (95% CI[0.1-0.6], P = 0.004). 22.3% (n = 4) eyes had persistent choroidal folds but BCVA was improved. Mean pre-intervention intraocular pressure (IOP) was 6.3 ± 3.8 mmHg (95% CI 4.4-8.2]) which increased to 12.1 ± 2.9 mmHg (95%CI[10.6-13.5], (P < 0.001). 27.8% (n = 5) eyes needed laser suture lysis post repair to control IOP; two needed further surgical intervention. Number of anti-glaucoma medications at last follow-up was 0.4 ± 0.9 (95% CI [-0.1-0.8], P = 0.09). No serious complications were encountered. Scleral patch graft and conjunctival advancement is a useful technique for repair of a scleral fistula post-filtering surgery, and this is recommended not only for the restoration of anatomy for prevention of infection and control of IOP, but also for visual rehabilitation.


Transposable elements in Drosophila.
Transposable elements (TEs) are mobile genetic elements that can mobilize within host genomes. As TEs comprise more than 40% of the human genome and are linked to numerous diseases, understanding their mechanisms of mobilization and regulation is important. Drosophila melanogaster is an ideal model organism for the study of eukaryotic TEs as its genome contains a diverse array of active TEs. TEs universally impact host genome size via transposition and deletion events, but may also adopt unique functional roles in host organisms. There are 2 main classes of TEs: DNA transposons and retrotransposons. These classes are further divided into subgroups of TEs with unique structural and functional characteristics, demonstrating the significant variability among these elements. Despite this variability, D. melanogaster and other eukaryotic organisms utilize conserved mechanisms to regulate TEs. This review focuses on the transposition mechanisms and regulatory pathways of TEs, and their functional roles in D. melanogaster.


Endoscopic Ultrasound in Pediatric Patients With Pancreatobiliary Disease.
Endoscopic ultrasound (EUS) is one of the greatest advances of the last few decades for the diagnostic and therapeutic approach of gastrointestinal diseases in adults. However, there are limitations to the application of the procedure in the pediatric population. The aim of this study was to describe the diagnostic yield and therapeutic role of EUS in pediatric patients with pancreatobiliary disease. A retrospective study was performed on a prospective database of pediatric patients who underwent diagnostic and therapeutic endosonographic procedures. All patients seen in a 12-year period were included. A total of 54 patients were included, including 32 (59.3%) female individuals, with an average age of 16 (9 to 17) years. EUS was abnormal in 46 (85%) patients. Of the abnormal procedures, 4 (7.4%) corresponded to therapeutic ones. The main indication of the study was recurrent acute pancreatitis in 29 (54%). The main endosonographic findings were microlithiasis in 14 (25.9%), chronic pancreatitis in 9 (16.7%), and pancreatic tumors in 6 (11.1%) patients. Follow-up was performed in 31 (57.4%) patients, 19/31 patients underwent surgery, and 4/31 patients had endoscopic retrograde cholangiopancreatography. The median follow-up was 910 (2 to 3916) days. In 100% of the patients with follow-up, the initial diagnosis of EUS was confirmed. EUS is a useful and safe tool in the pediatric population with pancreatobiliary diseases.


G-Quadruplex-Based Fluorescent Turn-On Ligands and Aptamers: From Development to Applications.
Guanine (G)-quadruplexes (G4s) are unique nucleic acid structures that are formed by stacked G-tetrads in G-rich DNA or RNA sequences. G4s have been reported to play significant roles in various cellular events in both macro- and micro-organisms. The identification and characterization of G4s can help to understand their different biological roles and potential applications in diagnosis and therapy. In addition to biophysical and biochemical methods to interrogate G4 formation, G4 fluorescent turn-on ligands can be used to target and visualize G4 formation both in vitro and in cells. Here, we review several representative classes of G4 fluorescent turn-on ligands in terms of their interaction mechanism and application perspectives. Interestingly, G4 structures are commonly identified in DNA and RNA aptamers against targets that include proteins and small molecules, which can be utilized as G4 tools for diverse applications. We therefore also summarize the recent development of G4-containing aptamers and highlight their applications in biosensing, bioimaging, and therapy. Moreover, we discuss the current challenges and future perspectives of G4 fluorescent turn-on ligands and G4-containing aptamers.


FGF7-like gene is associated with pericentric inversion of chromosome 9, and FGF7 is involved in the development of ovarian cancer.
Pericentric inversion of chromosome 9 is a common chromosomal aberration that may be involved in the pathogenesis of several types of cancer. Screening for structural balanced chromosomal aberrations in 58 patients with ovarian carcinoma found a patient with inv(9)(p11;q13) who had repeated spontaneous abortions. To define the breakpoint at the 9p11 region, the human 9p11-specific CEPH-YAC library was first screened with fluorescence in situ hybridization (FISH) analysis, resulting in isolation of the YAC clones 961d9 and 954b9 spanning the breakpoint. Next, screening with FISH analysis of the cosmid library constructed from 961D9 isolated 3 cosmid clones that included the breakpoint. DNA sequence analysis showed that the cosmid clones spanned the 57.7-kb sequence, which contained the FGF7 [keratinocyte growth factor (KGF)]-like gene including exons 2 and 3. FISH analysis showed that the 57.7-kb sequence was duplicated from the 9p11 region to the 9q13 region on the aberrant chromosome 9. Southern blot analysis showed multiple FGF7-like genes in the 9p11 region which were also duplicated to the 9q13 region on the aberrant chromosome 9. Because the FGF7-like genes are located at 9p11 and 9q13, our results are consistent with the hypothesis that the FGF7-like genes at 9p11 and 9p13 initiate the pericentric inversion of chromosome 9. Analysis of surgical specimens of ovarian cancer with the reverse transcription-polymerase chain reaction and immunohistochemical staining showed overexpression of the FGF7 gene in 4 of 9 stage I or II cases and in 10 of 11 stage III or IV cases. These findings suggest that FGF7 plays a significant role in the development of ovarian cancer.


Impact of routine open-mouth osseous temporomandibular joint tomography on diagnosis and therapeutic options.
The aim of this study was to evaluate the impact of routine open mouth osseous temporomandibular joint (TMJ) tomography on diagnosis and treatment planning of TMJ conditions. Investigating the need for such images is important to justify the additional radiation exposure imparted by such procedures. The records of patients who underwent closed- and open-mouth osseous TMJ tomography (planar film tomography up to the beginning of 2006; cone beam computed tomography from 2006 onward) were reviewed. Three examiners formulated a diagnosis and treatment plan based on the history, clinical findings, and tomography interpretation reports of closed-mouth images. Then they reviewed the interpretations of the open-mouth images and recorded whether they would change their original diagnosis and/or treatment plan on the basis of the findings. Descriptive analysis of the results was performed. The impact of routine open-mouth imaging on diagnosis and treatment planning was variable among the examiners. Examiners 1, 2, and 3 had a change in diagnosis in 7%, 5%, and 3% of cases, respectively, and a change in treatment plan in 1%, 1%, and 8%, respectively. Routine open-mouth osseous TMJ tomography had a minimal impact on diagnosis and treatment planning of TMJ conditions, which varied according to the treating clinician.


European longitudinal study on the relationship between adolescents' alcohol marketing exposure and alcohol use.
This is the first study to examine the effect of alcohol marketing exposure on adolescents' drinking in a cross-national context. The aim was to examine reciprocal processes between exposure to a wide range of alcohol marketing types and adolescent drinking, controlled for non-alcohol branded media exposure. Prospective observational study (11-12- and 14-17-month intervals), using a three-wave autoregressive cross-lagged model. School-based sample in 181 state-funded schools in Germany, Italy, Netherlands, Poland. A total of 9075 eligible respondents participated in the survey (mean age 14 years, 49.5% male. Adolescents reported their frequency of past-month drinking and binge drinking. Alcohol marketing exposure was measured by a latent variable with 13 items measuring exposure to online alcohol marketing, televised alcohol advertising, alcohol sport sponsorship, music event/festival sponsorship, ownership alcohol-branded promotional items, reception of free samples and exposure to price offers. Confounders were age, gender, education, country, internet use, exposure to non-alcohol sponsored football championships and television programmes without alcohol commercials. The analyses showed one-directional long-term effects of alcohol marketing exposure on drinking (exposure T1 on drinking T2: β = 0.420 (0.058), P < 0.001, 95% confidence interval (CI) = 0.324-0.515; exposure T2 on drinking T3: β = 0.200 (0.044), P < 0.001, 95% CI = 0.127-0.272; drinking T1 and drinking T2 on exposure: P > 0.05). Similar results were found in the binge drinking model (exposure T1 on binge T2: β = 0.409 (0.054), P < 0.001, 95% CI = 0.320-0.499; exposure T2 on binge T3: β = 0.168 (0.050), P = 0.001, 95% CI = 0.086-0.250; binge T1 and binge T2 on exposure: P > 0.05). There appears to be a one-way effect of alcohol marketing exposure on adolescents' alcohol use over time, which cannot be explained by either previous drinking or exposure to non-alcohol-branded marketing.


Differences between students' estimated and attained grades in a first-year introductory psychology course as a function of identity development.
Two hundred sixty-eight first-year university students were surveyed about the state of their identity development and their perceptions regarding chances for academic success in an introductory psychology course. In general, it was found that students who had an adult identity had a more accurate assessment of their chances for success in the course and also used more efficient study strategies. Students who had not completely formed an adult identity, however, were more inaccurate in estimates of their final grades and also seemed to use less productive study strategies. It was concluded that those who have formulated an adult identity might have also developed a more complete understanding of both themselves and their situation. Implications of the findings for further research regarding the effects of identity development on university life, as well as the implications for academic intervention programs, are discussed.


Copper(II) Binding Sites in N-Terminally Acetylated α-Synuclein: A Theoretical Rationalization.
The interactions between N-terminally acetylated α-synuclein and Cu(II) at several binding sites have been studied with DFT calculations, specifically with the M06 hybrid functional and the ωB97X-D DFT-D functional. In previous experimental studies, Cu(II) was shown to bind several α-synuclein residues, including Met1-Asp2 and His50, forming square planar coordination complexes. Also, it was determined that a low-affinity binding site exists in the C-terminal domain, centered on Asp121. However, in the N-terminally acetylated protein, present in vivo, the Met1 site is blocked. In this work, we simplify the representation of the protein by modeling each experimentally found binding site as a complex between an N-terminally acetylated α-synuclein dipeptide (or several independent residues) and a Cu(II) cation, and compare the results with a number of additional, structurally analogous sites not experimentally found. This way of representing the binding sites, although extremely simple, allows us to reproduce experimental results and to provide a theoretical rationale to explain the preference of Cu(II) for certain sites, as well as explicit geometrical structures for the complexes formed. These results are important to understand the interactions between α-synuclein and Cu(II), one of the factors inducing structural changes in the protein and leading to aggregated forms of it which may play a role in neurodegeneration.


Loop Integrands for Scattering Amplitudes from the Riemann Sphere.
The scattering equations on the Riemann sphere give rise to remarkable formulas for tree-level gauge theory and gravity amplitudes. Adamo, Casali, and Skinner conjectured a one-loop formula for supergravity amplitudes based on scattering equations on a torus. We use a residue theorem to transform this into a formula on the Riemann sphere. What emerges is a framework for loop integrands on the Riemann sphere that promises to have a wide application, based on off-shell scattering equations that depend on the loop momentum. We present new formulas, checked explicitly at low points, for supergravity and super-Yang-Mills amplitudes and for n-gon integrands at one loop. Finally, we show that the off-shell scattering equations naturally extend to arbitrary loop order, and we give a proposal for the all-loop integrands for supergravity and planar super-Yang-Mills theory.


Designing and evaluating healthcare ICT innovation: a cognitive engineering view.
Many healthcare ICT innovations fail in practice because of a failure to take into account user needs. User needs should be identified broadly, encompassing different levels of organization of the healthcare system and different stakeholder concerns. Full-scale simulators are starting to be used to help in the design and evaluation of novel biomedical devices and displays. Although promising, simulators have significant technical and operational limitations for this purpose and they do not address important aspects of the sociotechnical systems context in which healthcare ICT will be embedded. This argument is illustrated with a case study in which advanced auditory displays for patient monitoring were successfully evaluated in a full-scale patient simulator, but many further questions remain prior to successful translation to practice.


Propofol Target-controlled Infusion in Anesthesia Induction during Painless Gastroscopy.
To investigate the feasibility of using the eyelash reflex as an indicator to calculate the individualised optimal target concentration in anesthesia induction during painless gastroscopy. Experimental study. China-Japan Union Hospital of Jilin University, China, from January to December in 2016. A total of 180 patients, who were scheduled to receive painless fibergastroscopic examination or treatment in the last three months, were enrolled in this study. All patients were randomly divided into three groups, according to the doctor visiting order (n=60, each). During the induction of anesthesia using propofol target-controlled infusion, the effectsite concentration upon the disappearance of the eyelash reflex (C0) was recorded first. Then, one ug/kg of fentanyl was injected. At the same time, the target effect-site concentration induced by propofol was determined: the effect-site concentration in group A was 1.5 times of C0, the effect-site concentration in group B was two times of C0, and the effectsite concentration in group C was 2.5 times of C0. During anesthesia induction, the incidence of motor responses was higher in group A than in groups B and C (p<0.05), and the incidence of hypoxemia was significantly higher in group C than in groups A and B (p<0.01). In the anesthesia option of fentanyl combined with propofol target-controlled infusion, the effect-site concentration of propofol can be set to two times of that at the time the eyelash reflex disappears. This study provides a new pre-assessment method for the induction dose of propofol in painless gastroscopy.


The complete genome sequence of Bacillus velezensis strain GH1-13 reveals agriculturally beneficial properties and a unique plasmid.
The bacterial strain Bacillus velezensis GH1-13, isolated from rice paddy soil in Korea, has been shown to promote plant growth and have strong antagonistic activities against pathogens. Here, we report the complete genome sequence of GH1-13, revealing that it possesses a single 4,071,980-bp circular chromosome with 46.2% GC-content. The chromosome encodes 3,930 genes, and we have also identified a unique plasmid in the strain that encodes a further 104 genes (71,628bp and 31.7% GC-content). The genome was found to contain various enzyme-encoding operons, including indole-3-acetic acid (IAA) biosynthesis proteins, 2,3-butanediol dehydrogenase, various non-ribosomal peptide synthetases, and several polyketide synthases. These properties are responsible for the promotion of plant growth and the biosynthesis of secondary metabolites. They therefore have multiple beneficial effects that could be applied to agriculture. Through curing, we found that the unique plasmid of GH1-13 has important roles in the production of phytohormones, such as IAA, and in shaping phenotypic and physiological characteristics. The plasmid therefore likely influences the biological activities of GH1-13. The complete genome sequence of B. velezensis GH1-13 contributes to our understanding of this beneficial strain and will encourage research into its development for agricultural or biotechnological applications, enhancing productivity and crop quality.


Travel-Associated Venous Thromboembolism.
Long-distance travel is assumed to be a risk factor for venous thromboembolism (VTE). However, the available data have not clearly demonstrated the strength of this relationship, nor have they shown evidence for the role of thromboprophylaxis. We performed a systematic review of the literature. We also summarized available guidelines from 5 groups. We found 18 studies that addressed this question. Based on the data presented in the review, we conclude that there is an association between VTE and length of travel, but this association is mild to moderate in effect size with odds ratios between 1.1 and 4. A dose-response relationship between VTE and travel time was identified, with a 26% higher risk for every 2 h of air travel (P=0.005) starting after 4 h. The quality of evidence for both travel length and thromboprophylaxis was low. However, low-risk prophylactic measures such as graduated compression stockings were shown to be effective in VTE prevention. There is heterogeneity among the different practice guidelines. The guidelines generally concur that no prophylaxis is necessary in travelers without known thrombosis risk factors and advocate for conservative treatment such as compression stockings over pharmacologic prophylaxis. We conclude air travel is a risk factor for VTE and that there is a dose relationship starting at 4 h. For patients with risk factors, graduated compression stockings are effective prophylaxis.


'Ballroom itself can either make you or break you' - Black GBT Youths' psychosocial development in the House Ball Community.
Social context plays a significant role in adolescent identity development, particularly for youth lacking traditional systems of support. Using ecological and symbolic interactionism perspectives, this study qualitatively explored the psychosocial identity development of Black gay, bisexual, or transgendered youth participating in the House Ball Community (HBC). The HBC is a diverse network of family-like structures called 'houses', as well as a glamorous social outlet via pageant-like 'balls' in which participants compete. A series of focus groups were conducted with youth and leaders from the HBC (n = 37; age range = 17-24). Via cross-case and comparative analyses, specific motivating factors related to entry into and continued involvement in the community were identified. Factors related to entry into the community included lack of safe spaces, opportunities for acceptance, means of subsistence, and allure of the scene. Factors related to continued involvement included resilience and coping skills development, sexual identity acceptance and pride, prevalence of risky behaviour, and risk of exploitation. Discussion of these factors provides insight on how self-constructed, supplementary social contexts may provide both unique supports and risks to members, allowing for more focused and well-informed interventions and policies to enhance healthy development in such communities while mitigating risk.


IFN-β Improves Sepsis-related Alveolar Macrophage Dysfunction and Postseptic Acute Respiratory Distress Syndrome-related Mortality.
IFN-β is reported to improve survival in patients with acute respiratory distress syndrome (ARDS), possibly by preventing sepsis-induced immunosuppression, but its therapeutic nature in ARDS pathogenesis is poorly understood. We investigated the therapeutic effects of IFN-β for postseptic ARDS to better understand its pathogenesis in mice. Postseptic ARDS was reproduced in mice by cecal ligation and puncture to induce sepsis, followed 4 days later by intratracheal instillation of Pseudomonas aeruginosa to cause pneumonia with or without subcutaneous administration of IFN-β 1 day earlier. Sepsis induced prolonged increases in alveolar TNF-α and IL-10 concentrations and innate immune reprogramming; specifically, it reduced alveolar macrophage (AM) phagocytosis and KC (CXCL1) secretion. Ex vivo AM exposure to TNF-α or IL-10 duplicated cytokine release impairment. Compared with sepsis or pneumonia alone, pneumonia after sepsis was associated with blunted alveolar KC responses and reduced neutrophil recruitment into alveoli despite increased neutrophil burden in lungs (i.e., "incomplete alveolar neutrophil recruitment"), reduced bacterial clearance, increased lung injury, and markedly increased mortality. Importantly, IFN-β reversed the TNF-α/IL-10-mediated impairment of AM cytokine secretion in vitro, restored alveolar innate immune responsiveness in vivo, improved alveolar neutrophil recruitment and bacterial clearance, and consequently reduced the odds ratio for 7-day mortality by 85% (odds ratio, 0.15; 95% confidence interval, 0.03-0.82; P = 0.045). This mouse model of sequential sepsis → pneumonia infection revealed incomplete alveolar neutrophil recruitment as a novel pathogenic mechanism for postseptic ARDS, and systemic IFN-β improved survival by restoring the impaired function of AMs, mainly by recruiting neutrophils to alveoli.


Pulmonary artery pressure in lymphangioleiomyomatosis: an echocardiographic study.
Exercise-induced hypoxemia is frequent in patients with lymphangioleiomyomatosis (LAM) and could be associated with pulmonary hypertension. The aims of this study were to determine the prevalence of pulmonary hypertension in patients with LAM, to identify physiologic parameters associated with its occurrence, and to evaluate the effect of oxygen on response to exercise. Studies were performed in 120 patients. Complete data, including exercise echocardiography, pulmonary function testing, and standard cardiopulmonary exercise testing, were obtained in 95 patients. Resting pulmonary artery pressure (PAP) was 26+/-0.7 mm Hg (mean+/-SEM). Eight patients had pulmonary hypertension (43+/-3 mm Hg), and two patients had right ventricular dilatation. Ninety-five patients exercised (room air, n=64; oxygen, n=31) to a power of 58+/-2 W (49% of predicted) and an estimated peak oxygen uptake of 938+/-30 mL/min (56% of predicted). Sixty-one patients had a decline in arterial oxygen saturation (SaO2)>3%, and 56 patients had an elevation in PAP>40 mm Hg. Peak exercise PAP was negatively correlated with exercise Sao2 (p=0.0005). Multivariate analysis showed that exercise SaO2 was the best predictor of exercise PAP (p=0.012). Although resting pulmonary hypertension is rare in patients with LAM, a rise in PAP at low exercise levels occurs frequently, in part related to exercise-induced hypoxemia. Optimization of oxygen administration during activities of daily living should be undertaken in patients with LAM to prevent hypoxemia and exercise-induced pulmonary hypertension.


Biomechanics of the foot and ankle under static conditions.
The purpose of this article is to discuss the static mechanics of the foot and ankle. First, the motions of the ankle and foot available during nonambulatory activities are described by reviewing the literature discussing the axes of motion for the ankle and joints of the foot. Conflicting terminology is presented and clarified, and a scheme for a reasonable terminology is presented. The role of the ankle-foot complex in closed and open kinetic chains is also discussed. Terminology describing structural and functional positions of the foot is presented, including definitions of the subtalar neutral position. A systematic format of terminology is offered to reduce the current inconsistencies. Finally, the weight-bearing area of the foot and muscle activity in quiet standing are reviewed.


Characterization of cells resistant to the potent histone deacetylase inhibitor spiruchostatin B (SP-B) and effect of overexpressed p21waf1/cip1 on the SP-B resistance or susceptibility of human leukemia cells.
We previously showed that the B cell leukemia cell line NALM-6 had the highest susceptibility among a number of leukemia cell lines to spiruchostatin B (SP-B), a potent histone deacetylase (HDAC) inhibitor. We also showed that SP-B-induced cytotoxicity depended on induction of apoptosis that was mediated by p21waf1/cip1 expression. In the present study, we generated and characterized a stable, SP-B-resistant NALM-6 cell line (NALM-6/SP-B) by continuous exposure to SP-B, starting with a low SP-B concentration. NALM-6/SP-B cells were also more resistant to FK228, which has a similar chemical structure to SP-B, and were slightly more resistant to the P-gp substrates doxorubicin and vincristine than parental cells, but displayed similar susceptibility to other HDAC inhibitors and to paclitaxel as the parental cells. There was little change in the basal mRNA expression of HDAC1, p53, Bax, Bcl-2, Fas, caspase-3, c-Myc and MDR1 in NALM-6/SP-B compared to parental cells, but the mRNA expression of p21waf1/cip1 was decreased. The introduction of an exogenous p21waf1/cip1 expression vector restored SP-B induction of NALM-6/SP-B cell apoptosis. Moreover, overexpressed p21waf1/cip1 enhanced SP-B induction of the apoptosis of the human erythroleukemia leukemia cell line K562 which is less susceptible to SP-B than NALM-6 cells. These results suggest that downregulation of p21waf1/cip1, which is a characteristic feature of NALM-6/SP-B cells, was important for their resistance to SP-B, and that this SP-B resistance could be overcome by the introduction of exogenous p21waf1/cip1. Furthermore, introduction of p21waf1/cip1 to other leukemia cells such as K562 may enhance their susceptibility to SP-B. This is the first report of the characterization of SP-B-resistant cells and of the effect of overexpressed p21waf1/cip1 on the resistance or susceptibility of human leukemia cells to SP-B.


Acute and chronic exposure to shear stress have opposite effects on endothelial permeability to macromolecules.
Endothelial properties are affected by mechanical stresses. Several studies have shown that an acute application of shear stress increases the permeability of endothelial monolayers in culture. We investigated whether more prolonged application of shear has the opposite effect. Porcine aortic endothelial cells were cultured on Transwell filters to assess monolayer permeability to albumin. The medium above the cells was swirled using an orbital shaker; resultant shears were computed to lie within the physiological range. Acute application of shear increased permeability, but chronic application reduced it. The effect of chronic but not acute shear was reversed by inhibiting nitric oxide (NO) synthesis. The effect of chronic shear was also reversed by inhibiting phosphatidylinositol 3-OH kinase (PI3K) and soluble guanylyl cyclase. None of these interventions affected permeability under static conditions, and inhibition of cyclooxygenase was without effect. Chronic shear decreased mitosis rates by a fraction comparable to the reduction in permeability, but this effect was not reversed by inhibiting NO synthesis. We conclude that chronic application of shear stress reduces endothelial permeability to macromolecules by a PI3K-NO-cGMP-dependent mechanism. Since atherosclerosis can be triggered by excessive entry of plasma macromolecules into the arterial wall, the phenomenon may help explain the atheroprotective effects of shear and NO.