Quantitative volume and dynamic contrast-enhanced MRI derived perfusion of the infrapatellar fat pad in patellofemoral pain.
Patellofemoral pain (PFP) is a common knee condition and possible precursor of knee osteoarthritis (OA). Inflammation, leading to an increased perfusion, or increased volume of the infrapatellar fat pad (IPFP) may induce knee pain. The aim of the study was to compare quantitative dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) parameters, as imaging biomarkers of inflammation, and volume of the IPFP between patients with PFP and controls and between patients with and without IPFP edema or joint effusion. Patients with PFP and healthy controls were included and underwent non-fat suppressed 3D fast-spoiled gradient-echo (FSPGR) and DCE-MRI. Image registration was applied to correct for motion. The IPFP was delineated on FSPGR using Horos software. Volume was calculated and quantitative perfusion parameters were extracted by fitting extended Tofts' pharmacokinetic model. Differences in volume and DCE-MRI parameters between patients and controls were tested by linear regression analyses. IPFP edema and effusion were analyzed identically. Forty-three controls and 35 PFP patients were included. Mean IPFP volume was 26.04 (4.18) mL in control subjects and 27.52 (5.37) mL in patients. Median Ktrans was 0.017 (0.016) min-1 in control subjects and 0.016 (0.020) min-1 in patients. None of the differences in volume and perfusion parameters were statistically significant. Knees with effusion showed a higher perfusion of the IPFP compared to knees without effusion in patients only. The IPFP has been implicated as source of knee pain, but higher DCE-MR blood perfusion, an imaging biomarker of inflammation, and larger volume are not associated with PFP. Patient's knees with effusion showed a higher perfusion, pointing towards inflammation.


Chloroplast phosphoproteins. Evidence for a thylakoid-bound phosphoprotein phosphatase.
1. Isolated intact pea (Pisum sativum) chloroplasts incorporate [32P]orthophosphate into several thylakoid polypeptides in the light. Transfer of the labelled chloroplasts to darkness results in rapid dephosphorylation of the polypeptides. The most rapidly dephosphorylated phosphoproteins are the 26000-Mr doublet derived from the light-harvesting chlorophyll a/b binding complex. 2. Incubation of isolated 32P-labelled thylakoids in buffer in the absence of stromal components also results in rapid protein dephosphorylation. Again, the most rapidly dephosphorylated phosphoproteins are the 26000-Mr light-harvesting doublet. Dephosphorylation of all thylakoid phosphoproteins is accelerated by addition of up to 10 mM MgCl2. 3. The enzyme responsible for dephosphorylation is a phosphatase rather than a phosphotransferase or the thylakoid protein kinase acting in reverse. The enzyme is specifically and totally inhibited by NaF and does not require phosphoryl group acceptors such as ADP. Unlike the protein kinase, the phosphatase is indifferent to light and the electron transport inhibitor 3(3,4-dichlorophenyl)-1,1-dimethylurea. 4. The phosphorylated regions of the thylakoid phosphoproteins protrude from the outer surface of the membrane and are removed by trypsin treatment.


[Meniscectomy--total, partial, open or closed operation?].
The importance of the menisci for stability of the knee joint is pointed out. Because of its mechanical importance in the knee a more conservative behaviour for treatment is recommended. Even the degenerated meniscus can play his role sufficiently. For each type of lesion individual surgery is applied, especially if there is locking impingement linked with mechanically induced arthritis. Out of the clinical results follows that arthroscopic partial meniscectomy is superior to conventional open meniscectomy, especially in the early postoperative period.


High-throughput feature extraction for measuring attributes of deforming open-cell foams.
Metallic open-cell foams are promising structural materials with applications in multifunctional systems such as biomedical implants, energy absorbers in impact, noise mitigation, and batteries. There is a high demand for means to understand and correlate the design space of material performance metrics to the material structure in terms of attributes such as density, ligament and node properties, void sizes, and alignments. Currently, X-ray Computed Tomography (CT) scans of these materials are segmented either manually or with skeletonization approaches that may not accurately model the variety of shapes present in nodes and ligaments, especially irregularities that arise from manufacturing, image artifacts, or deterioration due to compression. In this paper, we present a new workflow for analysis of open-cell foams that combines a new density measurement to identify nodal structures, and topological approaches to identify ligament structures between them. Additionally, we provide automated measurement of foam properties. We demonstrate stable extraction of features and time-tracking in an image sequence of a foam being compressed. Our approach allows researchers to study larger and more complex foams than could previously be segmented only manually, and enables the high-throughput analysis needed to predict future foam performance.


[Distribution of free carboxyl groups in native pectins from fruits and vegetables].
Pectins from 18 kinds of fruit and vegetable are extracted with ammonium oxalate from the alcohol-insoluble substance and fractioned on DEAE-Sephacell. All the fruit pectins as well as tomato and sunflower pectin have in their distribution of the esterification degree a concentration maximum in the highly esterified range, that is above an esterification degree of 60%. With lower esterification degrees the concentration decreased continuously. Vegetable pectins from cucumbers, celery, and tomato-paprika (sweet, red) have their maximum within an esterification range of 50-60%. The carboxyl groups are randomly distributed in all these preparations. As against that, vegetable pectins from red and white cabbage, cauliflower, large radish and carrots have in addition to a pectin fraction with randomly distributed carboxyl groups another one, pectin molecules of which have blocks of free carboxyl groups. The latter fraction may constitute up to 50% of the pectin.


Disparities in outpatient and home health service utilization following stroke: results of a 9-year cohort study in Northern California.
To examine whether there are disparities in utilization of outpatient and home care services after stroke. Retrospective cohort study. The Kaiser Permanente of Northern California health care system, which provides health care for approximately 3.3 million members. A total of 11,119 patients hospitalized for a stroke between 1996 and 2003 and followed for 1 year. Receipt of outpatient rehabilitation (physical therapy, occupational therapy, speech pathology, or physical medicine and rehabilitation/physiatry visits), and/or home health care. There were significant differences in outpatient rehabilitation visits and home health enrollment during the year after acute care discharge for all the parameters under study. Older age and female gender were associated with less outpatient rehabilitation treatment, but these subpopulations were more likely to be enrolled in home health care. Non-whites, patients from urban areas, those with ischemic strokes, and those with longer acute care hospital stays had relatively more outpatient rehabilitation and were also more likely to be enrolled in the home health program. In addition, patients living in geographic areas with a median household income of $80,000 or more had significantly more outpatient rehabilitation visits than did patients living in lower income areas. Variations in outpatient rehabilitation visits and in home health care exist in this large integrated health system in terms of age, gender, race/ethnicity, residence area, type of stroke, and length of stay in an acute care hospital. The Kaiser Permanente integrated health care system seems to have outpatient stroke rehabilitation and home health programs that are providing care without disparities in relation to non-white populations, but other disparities appear to exist that may be related to socioeconomic factors, referral patterns, family support systems, or other cultural factors that have not been identified.


First-pass hepatic extraction and metabolic effects of insulin and insulin analogues.
First-pass hepatic extraction of insulin and hepatic and peripheral contributions to hypoglycemia were compared in conscious dogs during portal infusion of insulin A1, B29 diacetyl insulin, or A1-B29 dodecoyl insulin at 7 and 14 pmol X kg-1 X min-1. The liver removed 43 +/- 2% of insulin, 12 +/- 1% of dodecoyl, and 8 +/- 1% of diacetyl insulin, in a single transhepatic circulation. The hypoglycemia induced by insulin and diacetyl insulin and the ensuing glucagon response were greater than that produced by the dodecoyl analogue. Diacetyl insulin primarily increased glucose utilization, dodecoyl insulin solely inhibited hepatic production, and insulin affected both. The lack of hepatic effect of diacetyl insulin during hypoglycemia can be ascribed to greater counterregulation, because under euglycemic clamp conditions, this analogue caused suppression of glucose production. The different patterns of hypoglycemia exhibited can be explained by the combined effects of altered distribution between the liver and peripheral tissues caused by differences in hepatic extraction, the effect of this phenomenon on the counterregulatory response, and the intrinsic biological potency of the analogues.


Morphologically Tunable Rectangular Platelets Self-Assembled from Diblock Molecular Brushes Containing Azopyridine Pendants.
Two-dimensional (2D) platelet structures are of growing importance as building blocks for the preparation of optical and electrical devices. However, the creation of morphologically tunable rectangular platelets through polymer self-assembly still remains a challenge. Herein, we describe a rational strategy for the fabrication of 2D rectangular platelets by stacking azopyridine-containing diblock molecular brushes in two dimensions in a selective solvent. Amphiphilic PEG-co-(PtBA-g-PAzoPy) DMBs with poly(ethylene glycol) (PEG) block, poly(t-butyl acrylate) (PtBA) backbone, and poly(6-(4-(4-pyridyazo)phenoxy)-hexyl methacrylate) (PAzoPy) brush were synthesized by sequential reversible addition-fragmentation chain transfer polymerization and atom transfer radical polymerization. Various rectangular platelets were obtained via the solution self-assembly of PEG-co-(PtBA-g-PAzoPy) through a heating-cooling-aging process in which the morphology and size of platelets could be controlled by adjusting the composition of DMBs as well as the solvent polarity. In addition, we investigated the metal chelation ability and H-bonding-assisted co-assembly capability of PEG-co-(PtBA-g-PAzoPy). The results displayed that 2D hybrids and flower-like platelets were formed, respectively. Our study presents an efficient method to fabricate rectangular platelets with tunable morphologies.


[Screening questionnaire for documentation of medical history and diagnostic findings in dry eye disease].
Keratoconjunctivitis sicca is one of the most common ocular diseases world-wide. These patients suffer from severe symptoms which lead to an extremely reduced quality of life. Dry eye syndrome constitutes a major diagnostic and therapeutic challenge to all ophthalmologists because there is often a discrepancy between objective ocular signs and subjective symptoms of the patients. Furthermore, there exist only few causal therapeutic options. The physician-patient relationship plays an outstanding role in this condition. For the treatment of moderate to severe dry eye syndrome, special dry eye clinics have proved to be extremely useful. For follow-up measurements as well as the realisation of evidence-based medicine and quality control, it is a fundamental necessity to document symptoms, signs and therapy of these patients in order to optimise therapeutic strategies. For this purpose, we have developed special forms and standardised questionnaires for the individual documentation of medical history and diagnostic findings. To objectively assess the patient's complaints we use the "ocular surface disease index" (OSDI score). Only the establishment of standardised diagnostic and therapeutic algorithms with the help of special forms and questionnaires can help in the long run to improve the treatment of these severely affected patients.


Crepidotus crocophyllus found in Costa Rica and Mexico and revision of related species in subsection Fulvifibrillosi.
The study of Crepidotus specimens collected in Costa Rica and Mexico revealed that C. crocophyllus occurs in the tropical and subtropical forests of both countries. Type specimens of seven species related to C. crocophyllus in subsection Fulvifibrillosi s. Hesler and Smith were re-examined. Based on the morphological features, specimens supporting C. appalachianensis, C. aureifolius, C. distortus, C. subaureifolius and C. subnidulans are interpreted to be C. crocophyllus, thus all herein are proposed as its synonyms. Furthermore A. nephrodes is confirmed as a synonym of Crepidotus crocophyllus while Agaricus malachius, long considered contaxic with the former, is proposed as synonym of Crepidotus applanatus. The known records of C. crocophyllus indicate a wide but fragmented range of extension of the taxon throughout the Americas. Description, illustrations of microscopic features and discussions are provided.


A historical progression of the Midwest Regional Chapter of the SQA.
The Historical Committee of the Midwest Regional Chapter of the Society of Quality Assurance has recorded and tracked the evolution and historical progression of our chapter from the initial authorization of regional chapters by the national society in 1990, through its growth in membership and area served, to its current status in October 1996. This article describes this growth as well as meeting attendance, location and topics of the training seminars, membership distribution, and the significant milestones accomplished as the chapter journeyed through a whirlwind past.


Multistage carcinogenesis in the urinary bladder.
The induction of cancer of the urinary bladder is a multi-stage process involving multiple exogenous and endogenous factors. Based on the classical initiation-promotion model, we have used N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT) as initiator and sodium saccharin (SAC) or tryptophan as promoters. These latter chemicals have the properties expected of promoters: induction of hyperplasia, reversibility and nonmutagenicity. Also, tumors were induced whether the promoter was administered immediately after FANFT or beginning 6 weeks after FANFT was discontinued, but no tumors resulted if either promoter was given without initiation with FANFT. Factor(s) present in normal urine also are involved in the promotion process, in addition to the role of urine as a carrier of carcinogens. However, administration of SAC to animals with a rapidly proliferating bladder mucosa, induced by ulceration, pellet insertion, or in utero, resulted in bladder tumor induction, even without prior initiation with FANFT. To better understand the complex interaction of the multiple variables in bladder carcinogenesis, a stochastic computer model has been formulated based on long-term carcinogenicity and tissue kinetic studies in vivo. This model indicates the importance of cell proliferation and the development of hyperplasia in carcinogenesis.


The phospholipid and fatty acid composition of human platelet surface and intracellular membranes isolated by high voltage free flow electrophoresis.
Human platelets were labeled with tracer doses of [14C]arachidonic acid, then fractionated into mixed membranes which are separated into intracellular membranes and two different domains of surface membranes by high voltage free flow electrophoresis. Each subfraction was analyzed for its phospholipid content. Glycerophospholipids were separated by high performance liquid chromatography and their fatty acids analyzed by glass capillary gas chromatography. Intracellular membranes appeared substantially depleted in sphingomyelin, while enrichment of this phospholipid was seen in surface membranes. PC and PI were more enriched in intracellular membranes than in the surface membranes and the contrary was observed for PE. On the other hand, the pattern of the phospholipid labeling by [14C]arachidonate followed closely the glycerophospholipid profiles of the membrane subfractions, but the specific radioactivity of PI was higher than of PC, which itself was higher than that of PE. Moreover, the endogenous content of arachidonic acid accentuates these tendencies. The percentage of arachidonate in PE was higher in the surface membranes than in the intracellular membranes and the contrary was observed for arachidonyl-PC and PI. These differences were compensated for by certain saturated and monounsaturated fatty acids present in the composition profiles. These findings are discussed in relation to the membrane localization of lipases involved in the liberation of arachidonic acid for prostanoid synthesis.


[Changes in free thyroid hormones after epidural anesthesia].
The changes of FT4 and FT3 hormone levels were measured in 14 patients who were submitted to various light operations after epidural anesthesia. The FT3 changes were similar to those usually detected after general anesthesia, a fact that leads to the hypothesis that the afferent stimuli are not the only responsible for the thyroid hormone changes after stress.


XeF(2) coordination to a halogen center; Raman spectra (n = 1, 2) and X-ray crystal structures (n = 2) of [BrOF(2)][AsF(6)].nXeF(2) and [XOF(2)][AsF(6)] (X = Cl, Br).
The syntheses and structural characterizations of the [XOF(2)][AsF(6)] (X = Cl, Br) salts and the XeF(2) adduct-salts, [BrOF(2)][AsF(6)].nXeF(2) (n = 1, 2), are described. Although the [XOF(2)][AsF(6)] salts have been known for some time, their crystal structures had not been reported until the present study. The crystal structure of [BrOF(2)][AsF(6)] shows a positional disorder among the oxygen atom and the fluorine atoms. Both ClOF(2)(+) and BrOF(2)(+) have pseudo-octahedral coordination with a primary tripodal coordination sphere consisting of an oxygen atom and two fluorine atoms and a secondary coordination sphere consisting of three long contacts to fluorine atoms of different AsF(6)(-) anions. The low-temperature Raman spectra of [XOF(2)][AsF(6)] have been assigned on the basis of the crystal structures and with the aid of quantum-chemical calculations using [XOF(2)][AsF(6)](3)(2-) as a model for the crystallographic environment of XOF(2)(+). Several examples of XeF(2) coordinated through fluorine to transition metal centers are known, but no crystallographically characterized examples of XeF(2) coordinated to a nonmetal center other than xenon are known. The complex cation salts, [BrOF(2)][AsF(6)].nXeF(2) (n = 1, 2), were synthesized, and their Raman spectra have been assigned with the aid of quantum-chemical calculations. Although the structure of [BrOF(2)][AsF(6)].2XeF(2) is similar to that of the recently reported krypton analogue, notable differences occur. The contact distances between bromine and the fluorine atoms of NgF(2) (Ng = Kr, Xe) are shorter in [BrOF(2)][AsF(6)].2XeF(2) than in the KrF(2) analogue, which is attributed to the more polar natures of the Xe-F bonds. Unlike [BrOF(2)][AsF(6)].2KrF(2), which has been shown in the prior study to be stable in HF solution at room temperature, [BrOF(2)][AsF(6)].2XeF(2) enters into a dissociative equilibrium in which fluoride ion abstraction by BrOF(2)(+) occurs to give Xe(2)F(3)(+) and BrOF(3). The ELF and QTAIM analyses of [BrOF(2)][AsF(6)](3)(2-) and [BrOF(2)][AsF(6)].2XeF(2) were carried out and are compared with those of [BrOF(2)][AsF(6)].2KrF(2) and for free BrOF(2)(+) to better understand the effect of Br(V) coordination number on the localization domain of the Br(V) valence electron lone pair.


Warning signals and predator-prey coevolution.
Theories of the evolution of warning signals are typically expressed using analytic and computational models, most of which attribute aspects of predator psychology as the key factors facilitating the evolution of warning signals. Sherratt provides a novel and promising perspective with a model that considers the coevolution of predator and prey populations, showing how predators may develop a bias towards attacking cryptic prey in preference to conspicuous prey. Here, we replicate the model as an individual-based simulation and find, in accordance with Sherratt, that predators evolve a bias towards attacking cryptic prey. We then use a Monte Carlo simulation to calculate the relative survivorships of cryptic and conspicuous prey and stress that, as it stands, the model does not predict the evolution or stability of warning signals. We extend the model by giving predators continuous attack strategies and by allowing the evolution of prey conspicuousness: results are robust to the first modification but, in all cases, cryptic prey always enjoy a higher survivorship than conspicuous prey. When conspicuousness is allowed to evolve, prey quickly evolve towards crypsis, even when runaway coevolution is enabled. Sherratt's approach is promising, but other aspects of predator psychology, besides their innate response, remain vital to our understanding of warning signals.


Keep it local (and final): Remnant preferences in "let alone" ellipsis.
The let alone construction (John can't run a mile, let alone a marathon) differs from standard coordination structures (with and or but) by requiring ellipsis of the second conjunct--for example, a marathon is the remnant of an elided clause [[see text] a marathon]. In support of an ellipsis account, a corpus study of British and American English finds that let alone exhibits a Locality bias, as the second conjunct preferentially contrasts with the nearest lexical item of the same syntactic type. Two self-paced reading studies show that the Locality bias is active during online processing, but must be reconciled with indicators of semantic contrast and discourse information. Further, a sentence-rating study shows that the Locality bias interacts with a Finality bias that favours placing the let alone phrase at the end of a clause, which sometimes necessitates a nonlocal contrast. Together, the results show how a general bias in ellipsis for local contrasts is affected by discourse demands, such as the need for scalar contrast imposed by let alone, thereby offering a window into how possibly divergent syntactic and discourse constraints impact sentence processing.


[The functional properties of the cochlear-vestibular acupuncture points of the external auditory canal].
Endaural acupuncture points were studied in 60 healthy subjects. The points were found to have obvious advantages as compared with the commonly accepted points of the ear.


Vitamin D status during and after treatment and ovarian cancer survival.
Five-year relative survival for ovarian cancer remains below 50%. Strategies to improve outcomes are needed. Higher serum 25-hydroxyvitamin D [25(OH)D] concentrations [measure of vitamin D status] at and before diagnosis have been associated with longer survival in cancer patients; however, data for ovarian cancer are limited. We aimed to determine if 25(OH)D concentrations during and after primary treatment were associated with ovarian cancer-specific survival. We used data from a nationwide prospective cohort study of women with ovarian cancer. Among 886 participants treated with chemotherapy, 700 (79%) had a blood sample collected during (n = 591) and/or after (n = 458) primary treatment. These were tested for 25(OH)D. Clinical and survival data were abstracted from medical records. We used multivariable Cox proportional hazards regression to estimate hazard ratios (HR) and 95% confidence intervals (CI) for associations between 25(OH)D and ovarian cancer-specific survival. Mean 25(OH)D concentrations were lower during than after primary treatment (82 and 91 nmol/L, respectively); only 14% and 8% had concentrations below 50 nmol/L during and after primary treatment, respectively. There was no association between 25(OH)D and ovarian cancer-specific survival during five years of follow-up [HR 1.10 (95% CI: 0.76, 1.61) and 0.95 (0.54, 1.68) for the highest vs. lowest quintile during and after treatment, respectively]. We did not observe any association between serum 25(OH)D concentration and ovarian cancer-specific survival. Our results suggest that, in the absence of vitamin D deficiency, vitamin D supplementation to improve ovarian cancer survival is not warranted.


The Host Factor ANP32A Is Required for Influenza A Virus vRNA and cRNA Synthesis.
Influenza A viruses are negative-sense RNA viruses that rely on their own viral replication machinery to replicate and transcribe their segmented single-stranded RNA genome. The viral ribonucleoprotein complexes in which viral RNA is replicated consist of a nucleoprotein scaffold around which the RNA genome is wound, and a heterotrimeric RNA-dependent RNA polymerase that catalyzes viral replication. The RNA polymerase copies the viral RNA (vRNA) via a replicative intermediate, called the cRNA, and subsequently uses this cRNA to make more vRNA copies. To ensure that new cRNA and vRNA molecules are associated with ribonucleoproteins in which they can be amplified, the active RNA polymerase recruits a second polymerase to encapsidate the cRNA or vRNA. Host factor ANP32A has been shown to be essential for viral replication and to facilitate the formation of a dimer between viral RNA polymerases. Differences between mammalian and avian ANP32A proteins are sufficient to restrict viral replication. It has been proposed that ANP32A is only required for the synthesis of vRNA molecules from cRNA but not vice versa. However, this view does not match recent molecular evidence. Here we use minigenome assays, virus infections, and viral promoter mutations to demonstrate that ANP32A is essential for both vRNA and cRNA synthesis. Moreover, we show that ANP32A is not only needed for the actively replicating polymerase, but not for the polymerase that is encapsidating nascent viral RNA products. Overall, these results provide new insights into influenza A virus replication and host adaptation. IMPORTANCE Zoonotic avian influenza A viruses pose a constant threat to global health, and they have the potential to cause pandemics. Species variations in host factor ANP32A play a key role in supporting the activity of avian influenza A virus RNA polymerases in mammalian hosts. Here we show that ANP32A acts at two stages in the influenza A virus replication cycle, supporting recent structural experiments, in line with its essential role. Understanding how ANP32A supports viral RNA polymerase activity and how it supports avian polymerase function in mammalian hosts is important for understanding influenza A virus replication and the development of antiviral strategies against influenza A viruses.


Defective innate immune responses to respiratory syncytial virus infection in ovalbumin-sensitized mice.
Respiratory viral infections have frequently been reported to closely correlate with asthma exacerbations. Distinctive expression of cytokine/chemokine and anomalous responses of innate immunity induced by respiratory viral infections were suggested to play a key role. This study further evaluates the effects of airway sensitization on innate immunity in response to different viruses. Murine sensitization was established using an ovalbumin (OVA) sensitization model. Mice were subsequently infected with either respiratory syncytial virus (RSV) or human metapneumovirus (hMPV). Type I interferon (IFN), cytokines, and chemokines were measured in bronchoalveolar lavage (BAL) fluid. Pulmonary tissue samples were collected for the analysis of viral titers and type I IFN signal transcriptors. Distinct expressions of cytokine/chemokine responses after viral infection were also found in mice with OVA sensitization. A significant increase of virus replication was found in lungs of RSV-infected sensitized mice. The increment of RSV titer was associated with the decreased levels of type I IFN. Although Toll-like receptor 3 (TLR3) expression was significantly increased in the lungs, the key signal transcriptor, IFN regulatory factor 3, was significantly suppressed in the RSV-infected sensitized mice. A defective antiviral innate response was observed in the murine respiratory allergy model. Suppressed expression of IFN signal transcriptor contributes to decreased production of type I IFN. The defective innate immune response might result in acute viral exacerbations of allergic airway diseases.


Use of structural alterations in the synthesis of halothane metabolite antigens to mimic halothane-induced immunogen.
Four hapten-carrier conjugates were synthesized to evaluate any potential antigenic similarities between these synthetic compounds and the immunogens induced in vivo by the anesthetic, halothane and, thus, be used eventually as a more sensitive probe to detect the presence of these halothane-induced antibodies in halothane-exposed individuals. In this study, antibodies from five halothane hepatitis patients were used to evaluate these antigenic alterations since the specificity of these antibodies would most accurately reflect the antigenic structure of halothane-induced immunogens. Quantitation of antibody binding to these synthetic proteins was determined in an enzyme linked immunosorbent assay and immunoblot techniques. Trifluoroacetylated rabbit serum albumin was 5 times more reactive with these antibodies and thus more antigenic than the homologous acetylated moiety confirming the importance of the trifluoromethyl moiety as an epitope in the immunogen in vivo. Insertion of a spacer arm, aminocaproic acid, between the hapten and carrier moieties and an epitope density of 40% acetylation also increased antigenicity. Through these structural alterations produced in vitro, antigenic compounds have been produced which may resemble more closely the immunogen elicited in vivo and which may ultimately serve as more sensitive probes for halothane-induced antibodies from exposed individuals.


Golimumab in patients with active rheumatoid arthritis after treatment with tumor necrosis factor α inhibitors: findings with up to five years of treatment in the multicenter, randomized, double-blind, placebo-controlled, phase 3 GO-AFTER study.
The aim of this study was to assess long-term golimumab therapy in rheumatoid arthritis (RA) patients who discontinued previous tumor necrosis factor-α (TNF)-inhibitor(s). Patients enrolled into this multicenter, randomized, double-blind, placebo-controlled study of active RA (≥4 tender, ≥4 swollen joints) received placebo (Group 1) or golimumab 50 mg (Group 2) or 100 mg (Group 3) injections every 4 weeks. Patients in Groups 1 and 2 with inadequate response at week 16 escaped to golimumab 50 and 100 mg, respectively. At week 24, Group 1 patients crossed-over to golimumab 50 mg, Group 2 continued golimumab 50/100 mg per escape status, and Group 3 maintained dosing. During the long-term-extension (LTE), golimumab 50 mg could be increased to 100 mg, and 100 mg could be decreased to 50 mg. Data through 5 years are reported for all patients (safety) and patients using methotrexate (efficacy, intention-to-treat (ITT) analysis with last-observation-carried-forward for missing data and non-responder imputation for unsatisfactory efficacy discontinuations). In total, 459 of 461 randomized patients received the study agent, 304 of whom were methotrexate-treated and included in efficacy analyses. Through week 256, the proportions of methotrexate-treated patients achieving American-College-of-Rheumatology (ACR) responses were 37.6% to 47.0% for ACR20, 21.4% to 35.0% for ACR50, and 7.8% to 17.0% for ACR70 response across randomized groups. Golimumab safety through week 268 was generally consistent with that at week 24 and week 160 and other anti-TNF agents. In some patients with active RA discontinuing previous TNF-antagonist therapy, golimumab safety and efficacy, assessed conservatively with ITT analyses, was confirmed through 5 years. Clinicaltrials.gov NCT00299546 . Registered 03 March 2006.


An in situ growth approach to the fabrication of zeolite imidazolate framework-90 bonded capillary column for gas chromatography separation.
The unique properties of metal-organic frameworks, such as diversity in structures and pore sizes, high surface area, shape selectivity and available to functionality make them as potential materials of the stationary phase for gas chromatography. Here we show an in situ growth approach to the fabrication of zeolite imidazolate framework-90 (ZIF-90) bonded capillary column for gas chromatography separation. ZIF-90 was directly grown onto the inner wall of the carboxyl modified capillary via the coordination between Zn(II) and carboxyl group. The fabricated ZIF-90 bonded capillary column acted as a weak polar stationary phase. It not only exhibits high capacity in the separation of linear molecules, but also offers excellent features for the separation of 2- and 3-substituted ketones.


A pivotal role for antigen-presenting cells overexpressing SOCS3 in controlling invariant NKT cell responses during collagen-induced arthritis.
Suppressor of cytokine signalling (SOCS) proteins constitute a class of intracellular proteins that are key physiological regulators of immune cell function. It has previously been shown that antigen-presenting cells (APCs) overexpressing SOCS3 steer T helper immune responses and protect against experimental arthritis. A study was undertaken to investigate the contribution of SOCS3 in regulating invariant natural killer T (iNKT) cell responses during collagen-induced arthritis (CIA). DBA/1 mice were immunised with type II collagen and adenoviruses encoding SOCS3 were administered intravenously before the clinical onset of arthritis. Murine APCs overexpressing SOCS3 were co-cultured with an iNKT cell hybridoma and interleukin 2 (IL-2) release was measured by Luminex multi-analyte technology. The frequency and activation of primary iNKT cells was assessed by flow cytometry. Murine APCs were analysed for cytokine and CD1d expression following viral SOCS3 gene transfer. Viral overexpression of SOCS3 in APCs resulted in reduced activation of the iNKT cell hybridoma. Importantly, during initiation of CIA, adenovirus-mediated overexpression of SOCS3 in hepatic and splenic APCs inhibited iNKT cell expansion in both organs. The iNKT cell population from SOCS3-treated mice showed low expression of the early activation marker CD69 and primary liver iNKT cells produced less interferon γ and IL-4 upon α-galactosylceramide stimulation. No differences in CD1d surface expression were observed, but SOCS3-transduced APCs produced decreased levels of proinflammatory cytokines and increased levels of IL-10. These results demonstrate a critical role for SOCS3 in controlling the immunostimulatory capacities of APCs, which has direct implications for the effector function of iNKT cells during arthritis.


The amelioration of hepatic steatosis by thyroid hormone receptor agonists is insufficient to restore insulin sensitivity in ob/ob mice.
Thyroid hormone receptor (TR) agonists have been proposed as therapeutic agents to treat non-alcoholic fatty liver disease (NAFLD) and insulin resistance. We investigated the ability of the TR agonists GC-1 and KB2115 to reduce hepatic steatosis in ob/ob mice. Both compounds markedly reduced hepatic triglyceride levels and ameliorated hepatic steatosis. However, the amelioration of fatty liver was not sufficient to improve insulin sensitivity in these mice and reductions in hepatic triglycerides did not correlate with improvements in insulin sensitivity or glycemic control. Instead, the effects of TR activation on glycemia varied widely and were found to depend upon the time of treatment as well as the compound and dosage used. Lower doses of GC-1 were found to further impair glycemic control, while a higher dose of the same compound resulted in substantially improved glucose tolerance and insulin sensitivity, despite all doses being equally effective at reducing hepatic triglyceride levels. Improvements in glycemic control and insulin sensitivity were observed only in treatments that also increased body temperature, suggesting that the induction of thermogenesis may play a role in mediating these beneficial effects. These data illustrate that the relationship between TR activation and insulin sensitivity is complex and suggests that although TR agonists may have value in treating NAFLD, their effect on insulin sensitivity must also be considered.


Three-dimensional scanning transmission electron microscopy of dislocation loops in tungsten.
Scanning transmission electron microscopy (STEM) imaging using diffraction contrast is a powerful technique to assess crystal defects. In this work it is used to assess the spatial distribution of radiation induced defect in tungsten. In effect, its irradiation leads to the formation of nanometric dislocation loops that under certain conditions may form intriguing 3-D rafts. In this study, we have irradiated thin tungsten samples in situ in a TEM with 1.2 MeV W ions to 0.017 dpa at room temperature (RT) and at 700 °C. Besides the Burgers vector analysis, the number density and size of the dislocation loops with their spatial arrangement were quantitatively characterized by stereo imaging in STEM mode. Most of the loops have a Burgers vector ½ a0 〈111〉, with some a0 〈100〉 at room temperature. Loops are located mainly in the simulated damage profile but there is also a significant portion in deeper regions of the sample, indicating that loops in W diffuse easily, even at RT. At 700 °C, loops form elongated rafts that contain dislocation segments having a Burgers vector ½ a0 〈111〉. The rafts are narrow and reside on {111} planes; they are elongated along 〈110〉 directions, which correspond, when combined to the rafts' Burgers vector, to the lines of edge dislocations. Compared to conventional TEM, 3-D analysis in STEM appears thus as a powerful technique for quantitative analyses of defects in tungsten, as it allows reducing the background diffraction contrast and reaching thicker areas of the electron transparent foil, here 0.5 μm of tungsten at 200 kV.


Polo-like kinase inhibitor volasertib marginally enhances the efficacy of the novel Fc-engineered anti-CD33 antibody BI 836858 in acute myeloid leukemia.
Acute myeloid leukemia (AML) is the second most common type of leukemia in adults. Incidence of AML increases with age with a peak incidence at 67 years. Patients older than 60 years have an unfavorable prognosis due to resistance to conventional chemotherapy. Volasertib (BI 6727) is a cell-cycle regulator targeting polo-like kinase which has been evaluated in clinical trials in AML. We evaluated effects of volasertib in primary patient samples and NK cells. At equivalent doses, volasertib is cytotoxic to AML blasts but largely spares healthy NK cells. We then evaluated the effect of volasertib treatment in combination with BI 836858 on primary AML blast samples using antibody-dependent cellular cytotoxicity (ADCC) assays. Volasertib treatment of NK cells did not impair NK function as evidenced by comparable levels of BI 836858 mediated ADCC in both volasertib-treated and control-treated NK cells. In summary, volasertib is cytotoxic to AML blasts while sparing NK cell viability and function. Higher BI 836858 mediated ADCC was observed in patient samples pretreated with volasertib. These findings provide a strong rationale to test combination of BI 836858 and volasertib in AML.


Advanced single-slice rebinning in cone-beam spiral CT.
To achieve higher volume coverage at improved z-resolution in computed tomography (CT), systems with a large number of detector rows are demanded. However, handling an increased number of detector rows, as compared to today's four-slice scanners, requires to accounting for the cone geometry of the beams. Many so-called cone-beam reconstruction algorithms have been proposed during the last decade. None met all the requirements of the medical spiral cone-beam CT in regard to the need for high image quality, low patient dose and low reconstruction times. We therefore propose an approximate cone-beam algorithm which uses virtual reconstruction planes tilted to optimally fit 180 degrees spiral segments, i.e., the advanced single-slice rebinning (ASSR) algorithm. Our algorithm is a modification of the single-slice rebinning algorithm proposed by Noo et al. [Phys. Med. Biol. 44, 561-570 (1999)] since we use tilted reconstruction slices instead of transaxial slices to approximate the spiral path. Theoretical considerations as well as the reconstruction of simulated phantom data in comparison to the gold standard 180 degrees LI (single-slice spiral CT) were carried out. Image artifacts, z-resolution as well as noise levels were evaluated for all simulated scanners. Even for a high number of detector rows the artifact level in the reconstructed images remains comparable to that of 180 degrees LI. Multiplanar reformations of the Defrise phantom show none of the typical cone-beam artifacts usually appearing when going to larger cone angles. Image noise as well as the shape of the respective slice sensitivity profiles are equivalent to the single-slice spiral reconstruction, z-resolution is slightly decreased. The ASSR has the potential to become a practical tool for medical spiral cone-beam CT. Its computational complexity lies in the order of standard single-slice CT and it allows to use available 2D backprojection hardware.


Improved Stability of Human CGI-58 Induced by Phosphomimetic S237E Mutation.
In lipolysis, the activating function of CGI-58 is regulated by its interaction with perilipin 1 (PLIN1) localized on the lipid droplet (LD), and its release is controlled by phosphorylation. Once lipolysis is stimulated by catecholamines, protein kinase A (PKA)-mediated phosphorylation enables the dissociation of the CGI-58/PLIN1 complex, thereby recruiting adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) to initiate fatty acid release. It has been shown that mouse CGI-58 mutant S239E, which mimics the phosphorylation of this residue, is able to dissociate from the CGI-58/PLIN1 complex and activate ATGL. Here, we analyze the stabilizing effect on human CGI-58 of a triple tryptophan to alanine mutant (3WA) on the LD-binding motif, as well as a quadruple mutant in which the phosphomimetic S237E substitution was introduced to the 3WA construct (3WA/S237E). We found that tryptophan residues promote wild-type (WT) protein aggregation in solution since their substitution for alanine residues favors the presence of the monomer. Our experimental data showed increased thermal stability and solubility of 3WA/S237E protein compared to the 3WA mutant. Moreover, the 3WA/S237E protein showed proper folding and a functional binding site for oleoyl-CoA. The analysis of a bioinformatic three-dimensional (3D) model suggests an intramolecular interaction between the phosphomimetic glutamic acid and a residue of the α/β hydrolase core. This could explain the increased solubility and stability observed in the 3WA/S237E mutant and evidences the possible role of serine 237 phosphorylation.


The DNA methylome of inflammatory bowel disease (IBD) reflects intrinsic and extrinsic factors in intestinal mucosal cells.
Abnormal DNA methylation has been described in human inflammatory conditions of the gastrointestinal tract, such as inflammatory bowel disease (IBD). As other complex diseases, IBD results from the balance between genetic predisposition and environmental exposures. As such, DNA methylation may be the consequence (and potential effector) of both, genetic susceptibility variants and/or environmental signals such as cytokine exposure. We attempted to discern between these two non-excluding possibilities by performing a combined analysis of published DNA methylation data in intestinal mucosal cells of IBD and control samples. We identified abnormal DNA methylation at different levels: deviation from mean methylation signals at site and region levels, and differential variability. A fraction of such changes is associated with genetic polymorphisms linked to IBD susceptibility. In addition, by comparing with another intestinal inflammatory condition (i.e., coeliac disease) we propose that aberrant DNA methylation can also be the result of unspecific processes such as chronic inflammation. Our characterization suggests that IBD methylomes combine intrinsic and extrinsic responses in intestinal mucosal cells, and could point to knowledge-based biomarkers of IBD detection and progression.


[Urolithiasis and pregnancy].
Urolithiasis disease during pregnancy fortunately is a relatively rare disease; nevertheless many times it complicates the diagnosis and treatment. This paper reviews the safety and efficacy of both and their relationships with mother and fetus and proposes different options for the urologist, gynecologist, and urogynecologist for the attention of pregnant women with suspicion of urolithiasis. The ultrasound seems to be the first-choice method during the pregnancy; it is available in many health centers and does not require too much experience for its interpretation. Intravenous urography, simple X-ray and magnetic resonance image with its limitations aid support to our clinical suspicion; additionally the uretheroscopy is the diagnostic and therapeutic method and can be used safely during the pregnancy; thus first-choice study is the ultrasound with changes in vascular resistance rates. Once diagnosis is established, conservative treatment is applied because the majority of patients has spontaneous elimination of stones (70 to 80%). Surgical intervention can be an option and when is needed, placement of an uretheral catheter JJ, percutaneous nephrostomy and uretheroscopy may be an option to definitive treatment.


[Postoperative nutrition in patients with head and neck cancer].
Patients who underwent surgery for head and neck malignant neoplasms are problematic because dysphagia, pain and postoperative secuelaes. Nutritional support is necessary in more than 90% of all patients with head and neck cancer. The purpose of this study is to determine nutritional support in these patients and problems related to nutrition. Fifty-four patients with oral and pharyngolaryngeal carcinomas underwent surgery were studied prospectively. We studied nutritional support (oral nutrition, enteral nutrition or parenteral nutrition) and problems related to nutrition: aspiration neumoniae and pharyngostoma. The 7% of the patients underwent oral feeding after surgery, 87% enteral nutrition with nasogastric tube and 6% parenteral nutrition. We identified neumoniae in 6% of patients and faringocutaneous fistula in 19%. When patients leaved the hospital, 98% of the patients underwent oral feeding and one patient had nasopharyngeal tube for enteral nutrition. 1. Artificial nutrition was necessary in 93% patients with oropharyngolaryngealcancer; 2. Enteral nutrition was the most useful modality of alimentation (87%); 3.25% of patients had postoperative complications: (18% pharyngostoma, 6% neumoniae); 4. Oral feeding was possible in 98% of patients out of hospital.


Risk of self-harm in patients with eating disorders: English population-based national record-linkage study, 1999-2021.
Anorexia nervosa (AN) and bulimia nervosa (BN) are eating disorders associated with high rates of self-harm (SH). This is the first national study in England to quantify this association in a hospital population. A retrospective cohort study using a linked national dataset of Hospital Episode Statistics for 1999-2021. The exposure cohort included individuals aged <35 years admitted to hospital with a diagnosis of AN or BN. The reference cohort included hospital controls. We calculated the rate ratio (RR) of SH in each cohort. The individuals in the two cohorts were matched on multiple socio-demographic indicators. The main outcome was a subsequent hospitalization or death record with an SH diagnosis. We identified 15,004 females and 1411 males with AN, and 6055 females and 741 males with BN. The RR with 95% confidence intervals (95%CI) for a subsequent admission with intentional self-harm after admission with AN was 4.9 (95%CI 4.7-5.1) in females and 4.8 (95%CI 3.9-5.8) in males. For BN it was 9.0 (95%CI 8.4-9.6) in females and 9.8 (95%CI 7.7-12.2) in males. There were strong associations between AN and BN and other SH. Women and men admitted to English hospitals with AN or BN have a very high risk of a subsequent admission with SH. For some SH behaviors, such as alcohol intoxication, the RR was >10-fold elevated. The magnitude of risk was higher for BN than for AN. Clinicians should be aware of the scale of risk increase. Providing those at risk with appropriate support is required. This study is the first national study in an English hospital population that confirms and quantifies the association between eating disorders and self-harm. We have found that both women and men admitted to hospital with anorexia nervosa or bulimia nervosa are at an increased risk of subsequent admission with self-harm. It is important that clinicians are aware of this increased risk to support those at highest risk of self-harm.


Dietary flaxseed protects against ventricular fibrillation induced by ischemia-reperfusion in normal and hypercholesterolemic Rabbits.
Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), the (n-3) PUFA found in fish oils, exert antiarrhythmic effects during ischemia. Flaxseed is the richest plant source of another (n-3) PUFA, alpha-linolenic acid (ALA), yet its effects remain largely unknown. Our objective was to determine whether a flaxseed-rich diet is antiarrhythmic in normal and hypercholesterolemic rabbits. Male New Zealand White (NZW) rabbits (n = 14-16) were fed as follows: regular diet (REG group); diet containing 10% flaxseed (FLX group); 0.5% cholesterol (CHL group); or 0.5% cholesterol + 10% flaxseed (CHL/FLX group) for up to 16 wk. Plasma cholesterol was significantly elevated in the CHL and CHL/FLX groups. Plasma triglycerides were unchanged. ALA levels increased significantly in plasma and hearts of the FLX and CHL/FLX groups. After the feeding period, rabbit hearts were isolated and subjected to global ischemia (30 min) and reperfusion (45 min). Ventricular fibrillation (VF) occurred during ischemia in 33% of REG but in none of FLX hearts, and 28% of CHL but only 6% of CHL/FLX hearts. VF incidence during reperfusion was 28% and 26% in REG and FLX hearts, respectively. The incidence significantly increased to 64% in CHL hearts, and was significantly attenuated (18%) in CHL/FLX hearts. CHL markedly prolonged the QT interval, whereas FLX significantly shortened the QT interval and reduced arrhythmias in the FLX and CHL/FLX hearts. In vitro application of (n-3) PUFA shortened the action potential duration, an effect consistent with the QT data. This study demonstrates that dietary flaxseed exerts antiarrhythmic effects during ischemia-reperfusion in rabbit hearts, possibly through shortening of the action potential.


Revealing internal flow behaviour in arc welding and additive manufacturing of metals.
Internal flow behaviour during melt-pool-based metal manufacturing remains unclear and hinders progression to process optimisation. In this contribution, we present direct time-resolved imaging of melt pool flow dynamics from a high-energy synchrotron radiation experiment. We track internal flow streams during arc welding of steel and measure instantaneous flow velocities ranging from 0.1 m s-1 to 0.5 m s-1. When the temperature-dependent surface tension coefficient is negative, bulk turbulence is the main flow mechanism and the critical velocity for surface turbulence is below the limits identified in previous theoretical studies. When the alloy exhibits a positive temperature-dependent surface tension coefficient, surface turbulence occurs and derisory oxides can be entrapped within the subsequent solid as result of higher flow velocities. The widely used arc welding and the emerging arc additive manufacturing routes can be optimised by controlling internal melt flow through adjusting surface active elements.


Physical therapy in patients with complex regional pain syndrome type I after distal radius fracture: a case series.
[Purpose] To describe the effect of a physical therapy program in function improvement and pain reduction in patients older than 60 years with complex regional pain syndrome (CRPS) type I after distal radius fracture (DRF) treated conservatively. [Participants and Methods] Fifty-four patients received a 6 weeks physical therapy program that included in hydrotherapy, manual therapy, and exercises based on motor skill training. Two evaluations were performed, the wrist/hand function was assessed with Patient-Rated Wrist Evaluation (PRWE) questionnaire, the upper extremity function with the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire, grip strength with Jamar Dynamometer, and pain intensity with the Visual Analog Scale (VAS). [Results] At the end of the treatment, PRWE showed a decrease of 30.9 points, DASH 34.7 points, and the VAS, 3.4 cm. The grip strength showed an increase of 14.4%. [Conclusion] A physical therapy program based on hydrotherapy, manual therapy, and exercises in a short term improves the function and reduces the pain in patients older than 60 years with CRPS I after DRF treated conservatively.


Integrative Self-assembly of Graphene Quantum Dot and Biopolymer into a Versatile Biosensing Toolkit.
Hybrid self-assembly has become a reliable approach to synthesize soft materials with multiple levels of structural complexity and synergistic functionality. In this work, photoluminescent graphene quantum dots (GQDs, 2-5 nm) are used for the first time as molecule-like building blocks to construct self-assembled hybrid materials for label-free biosensors. Ionic self-assembly of disc-shaped GQDs and charged biopolymers is found to generate a series of hierarchical structures that exhibit aggregation-induced fluorescence quenching of the GQDs and change the protein/polypeptide secondary structure. The integration of GQDs and biopolymers via self-assembly offers a flexible toolkit for the design of label-free biosensors in which the GQDs serve as a fluorescent probe and the biopolymers provide biological function. The versatility of this approach is demonstrated in the detection of glycosaminoglycans (GAGs), pH, and proteases using three strategies: 1) competitive binding of GAGs to biopolymers, 2) pH-responsive structural changes of polypeptides, and 3) enzymatic hydrolysis of the protein backbone, respectively. It is anticipated that the integrative self-assembly of biomolecules and GQDs will open up new avenues for the design of multifunctional biomaterials with combined optoelectronic properties and biological applications.


A Critical Review of the Current Global Ex Situ Conservation System for Plant Agrobiodiversity. I. History of the Development of the Global System in the Context of the Political/Legal Framework and Its Major Conservation Components.
The history of ex situ conservation is relatively short, not more than a century old. During the middle of last century, triggered by the realization that genetic erosion was threatening the existing landraces and wild relatives of the major food crops, global efforts to collect and conserve the genetic diversity of these threatened resources were initiated, predominantly orchestrated by FAO. National and international genebanks were established to store and maintain germplasm materials, conservation methodologies were created, standards developed, and coordinating efforts were put in place to ensure effective and efficient approaches and collaboration. In the spontaneously developing global conservation system, plant breeders played an important role, aiming at the availability of genetic diversity in their breeding work. Furthermore, long-term conservation and the safety of the collected materials were the other two overriding criteria that led to the emerging international network of ex situ base collections. The political framework for the conservation of plant genetic resources finds its roots in the International Undertaking of the FAO and became 'turbulent rapid' with the conclusion of the Convention on Biological Diversity. This paper reviews the history of the global ex situ conservation system with a focus on the international network of base collections. It assesses the major ex situ conservation approaches and methods with their strengths and weaknesses with respect to the global conservation system and highlights the importance of combining in situ and ex situ conservation.


Influence of unsaturated fatty acid membrane component on sensitivity of an Escherichia coli fatty acid auxotroph to conditions of nutrient depletion.
The unsaturated fatty acid auxotroph Escherichia coli AK7 was provided with either oleic acid (cis 18:1) or linolenic acid (cis 18:3) to vary the degree of unsaturation of cell membrane lipids. The susceptibility of oleic acid- and linolenic acid-grown cells to starvation at 37 degrees C in 154 mM NaCl was compared following the decline in the number of CFU by plating the cells on agar medium. The decline in CFU was faster for linolenic acid-than for oleic acid-grown cells, but it was not indicative of cell death, since culturable CFU was recovered after respirable substrate was added to the starved cell suspension. Cell envelope microviscosity (determined by fluorescence polarization) of oleic acid- and linolenic acid-grown cells was equal in the presence of a respirable substrate, but in its absence the microviscosity of linolenic acid-grown cells was lower than that of oleic acid-grown cells. The results suggest that cell envelope microviscosity is an important factor in determining the sensitivity of E. coli to conditions of nutrient depletion.


Reprogrammed mouse fibroblasts differentiate into cells of the cardiovascular and hematopoietic lineages.
Forced expression of the four transcription factors Oct4, Sox2, c-Myc, and Klf4 is sufficient to confer a pluripotent state upon the murine fibroblast genome, generating induced pluripotent stem (iPS) cells. Although the differentiation potential of these cells is thought to be equivalent to that of embryonic stem (ES) cells, it has not been rigorously determined. In this study, we sought to identify the capacity of iPS cells to differentiate into Flk1-positive progenitors and their mesodermal progeny, including cells of the cardiovascular and hematopoietic lineages. Immunostaining of tissues from iPS cell-derived chimeric mice demonstrated that iPS cells could contribute in vivo to cardiomyocytes, smooth muscle cells, endothelial cells, and hematopoietic cells. To compare the in vitro differentiation potential of murine ES and iPS cells, we either induced embryoid body (EB) formation of each cell type or cultured the cells on collagen type IV (ColIV), an extracellular matrix protein that had been reported to direct murine ES cell differentiation to mesodermal lineages. EB formation and exposure to ColIV both induced iPS cell differentiation into cells that expressed cardiovascular and hematopoietic markers. To determine whether ColIV-differentiated iPS cells contained a progenitor cell with cardiovascular and hematopoietic differentiation potential, Flk1-positive cells were isolated by magnetic cell sorting and exposed to specific differentiation conditions, which induced differentiation into functional cardiomyocytes, smooth muscle cells, endothelial cells, and hematopoietic cells. Our data demonstrate that murine iPS cells, like ES cells, can differentiate into cells of the cardiovascular and hematopoietic lineages and therefore may represent a valuable cell source for applications in regenerative medicine. Disclosure of potential conflicts of interest is found at the end of this article.


Driving under the influence among frequent ecstasy consumers in Australia: trends over time and the role of risk perceptions.
Driving under the influence (DUI) of alcohol and illicit drugs is a serious road safety concern. This research aimed to examine trends in DUI across time and changes in attitudes towards the risks (crash and legal) associated with DUI among regular ecstasy users (REU) interviewed in Australia. Participants were regular (at least monthly) ecstasy users surveyed in 2007 (n=573) or 2011 (n=429) who had driven a car in the last six months. Face to face interviews comprised questions about recent engagement of DUI and roadside breath (alcohol) and saliva (drug) testing. Participants also reported the risk of crash and of being apprehended by police if DUI of alcohol, cannabis, ecstasy, and methamphetamine. There were significant reductions in DUI of psychostimulants (ecstasy, methamphetamine, cocaine, LSD) but not alcohol or cannabis between 2007 and 2011. This was accompanied by increased experience of roadside saliva testing and increases in crash and legal risk perceptions for ecstasy and methamphetamine, but not alcohol or cannabis. When the relationship between DUI and risk variables was examined, low crash risk perceptions were associated with DUI of all substances and low legal risk perceptions were associated with DUI of ecstasy. The observed reduction in DUI of psychostimulants among frequent ecstasy consumers may be related to increased risk awareness stemming from educational campaigns and the introduction of saliva testing on Australian roads. Such countermeasures may be less effective in relation to deterring or changing attitudes towards DUI of cannabis and alcohol among this group.


Diagnosis of macular pseudoholes and lamellar macular holes: is optical coherence tomography the "gold standard"?
To assess fundus autofluorescence (AF) for differential diagnosis of macular pseudoholes (MPH) and lamellar macular holes (LMH) evaluated by optical coherence tomography (OCT) as the "gold standard". The files on 50 eyes of 46 consecutive patients diagnosed by OCT as having a foveal defect with residual retinal tissue at the bottom were reviewed. Retinal thickness was measured at the foveal centre and 750 microm temporally and nasally to differentiate further MPH and LMH. The corresponding corrected AF images were then evaluated. Eyes with either macular pucker or stage 1a impending macular hole served as controls. OCT measurements allowed the classification of two different profiles: 28 eyes classified with MPH had macular centres and perifoveal retinas that were significantly thicker than the 22 eyes classified with LMH. The corrected value of the foveal AF intensity was not significantly different between the two groups. In addition, the AF did not correlate with the thickness of the retinal tissue at the base of either MPH or LMH eyes. None of the control eyes showed foveal AF. The findings suggest that OCT data must be interpreted with caution when differentiating between MPH and LMH. In this series, the two groups showed similar foveal AF. AF imaging may add useful information to the differential diagnosis of MPH from LMH: the presence of foveal AF is consistent with a loss of foveal tissue and therefore a diagnosis of LMH.


Processing methods of the pipeline crack detection signal by a balanced field electromagnetic technique based on phase characteristics.
The balanced field electromagnetic technique is an effective way of in-line inspection to detect cracks in pipelines. A signal demodulation method based on phase characteristics is proposed for the problem of interference signals generated by the sensor tilt shaking during the detection, which affects the judgment of the cracks. The method uses a reference signal whose phase is orthogonal to the signal generated by the sensor shaking to demodulate the detection signal to eliminate the shake interference. The generation principles of crack detection signals and interference signals generated by sensor shaking are analyzed, and the influence of sensor lift-off on detection is compared. A demodulation model is established based on the characteristic of that same frequency and different phases of crack and shake signals. The feasible conditions of the method are analyzed by simulation, and the phase value of the reference signal in the demodulation method is determined. The platform detection experiment and pulling tests at different speeds are carried out, respectively, to verify the effectiveness of the proposed method. The results show that there is a significant phase difference between the signals generated by the sensor shaking and the crack. For carbon steel pipelines, the signal phase of different shake angles is -4°. When the sensor structure and excitation frequency in this study are used, the reference signal phase is chosen to be 86°. The method preserves the detection signal characteristics before processing and enables the linear output responses to be obtained for different depths of cracks.


Neuroimaging of Central Sensitivity Syndromes: Key Insights from the Scientific Literature.
Central sensitivity syndromes are characterized by distressing symptoms, such as pain and fatigue, in the absence of clinically obvious pathology. The scientific underpinnings of these disorders are not currently known. Modern neuroimaging techniques promise new insights into mechanisms mediating these postulated syndromes. We review the results of neuroimaging applied to five central sensitivity syndromes: fibromyalgia, chronic fatigue syndrome, irritable bowel syndrome, temporomandibular joint disorder, and vulvodynia syndrome. Neuroimaging studies of basal metabolism, anatomic constitution, molecular constituents, evoked neural activity, and treatment effect are compared across all of these syndromes. Evoked sensory paradigms reveal sensory augmentation to both painful and nonpainful stimulation. This is a transformative observation for these syndromes, which were historically considered to be completely of hysterical or feigned in origin. However, whether sensory augmentation represents the cause of these syndromes, a predisposing factor, an endophenotype, or an epiphenomenon cannot be discerned from the current literature. Further, the result from cross-sectional neuroimaging studies of basal activity, anatomy, and molecular constituency are extremely heterogeneous within and between the syndromes. A defining neuroimaging "signature" cannot be discerned for any of the particular syndromes or for an over-arching central sensitization mechanism common to all of the syndromes. Several issues confound initial attempts to meaningfully measure treatment effects in these syndromes. At this time, the existence of "central sensitivity syndromes" is based more soundly on clinical and epidemiological evidence. A coherent picture of a "central sensitization" mechanism that bridges across all of these syndromes does not emerge from the existing scientific evidence.


Matrix metalloproteinases and the regulation of tissue remodelling.
Matrix metalloproteinases (MMPs) were discovered because of their role in amphibian metamorphosis, yet they have attracted more attention because of their roles in disease. Despite intensive scrutiny in vitro, in cell culture and in animal models, the normal physiological roles of these extracellular proteases have been elusive. Recent studies in mice and flies point to essential roles of MMPs as mediators of change and physical adaptation in tissues, whether developmentally regulated, environmentally induced or disease associated.


Suppression of growth hormone release restores phosphaturic response to PTH in immature rats.
Immature rats display a blunted rise in urinary phosphate but not adenosine 3',5'-cyclic monophosphate (cAMP) excretion in response to parathyroid hormone (PTH), perhaps as a consequence of the increased demand for phosphate during growth. Because a major driving force for growth is growth hormone (GH), and in view of the fact that GH has been shown to promote renal phosphate retention in the immature animal, it is possible that GH may attenuate the phosphaturic effect of PTH. The objective of this study was to determine whether suppression of pulsatile GH release, during administration of a synthetic peptide antagonist to GH-releasing factor, i.e., [N-acetyl-Tyr1-D-Arg2]-GRF-(1-29)-NH2 (GRF-AN), alters the renal response to increasing doses of PTH (1.5-15.0 micrograms.100 g-1.h-1) in the acutely thyroparathyroidectomized immature rat. Baseline fractional excretion of phosphate (FEPi), before administration of PTH, was negligible in all groups (less than 0.05%). Infusion of PTH resulted in an attenuated rise in FEPi in immature control rats compared with adult control rats (from 3.8 +/- 1.4% at lowest PTH dose to 16.7 +/- 3.1% at highest dose in immature rats vs. 21.1 +/- 3.5 to 31.9 +/- 4.4% in adult rats, P less than 0.05). In contrast, immature rats treated for 2 days with GRF-AN (100 micrograms/kg, twice daily) displayed an enhanced phosphaturic response (FEPi from 12.0 +/- 4.2 to 42.9 +/- 3.7%, P less than 0.05) compared with immature control rats, which was not different from that observed in control adult rats.(ABSTRACT TRUNCATED AT 250 WORDS)


A simplified technique for treating the complex dislocation of the index metacarpophalangeal joint.
The volar approach to open reduction of the complex dislocation of the index metacarpophalangeal joint as described by Kaplan proved to have certain disadvantages. Digital nerves are easily damaged during exposure and there is a limited view of the entrapped fibrocartilaginous volar plate dorsal to the metarcarpal head. A direct dorsal longitudinal incision through the skin and extensor tendon gives full exposure. The volar plate attached to the proximal phalanx and trapped over the dorsal aspect of the metacarpal head is in full view. The volar plate is split longitudinally and the dislocation reduces spontaneously as the flexor tendons and lumbrical muscle slip by the metacarpal head. The advantages of this approach as compared with the volar approach are: (1) there is full exposure of the fibrocartilaginous volar plate, the main structure blocking reduction; (2) digital nerves are not as apt to be damaged; and (3) accurate reduction and fixation of the osteochondral fracture of the metacarpal head, frequently seen with this dislocation, is possible.


Barriers to effective conversations regarding overweight and obesity in regional Victoria.
General practitioners (GPs) have a crucial role to play in engaging patients in discussions about overweight and obesity. However, such discussions are currently uncommon. The aim of this study was to examine how GPs in rural areas talk about overweight and obesity with their patients, specifically to identify key barriers to effective conversations. This study used a qualitative methodology. Semi-structured interviews were conducted with GPs (n = 7) and patients (n = 7) across two rural areas. Key barriers to effective conversations between GPs and patients about overweight and/or obesity include: uncertainty about appropriate language; lack of time; concerns about compromising mutual trust and rapport; concerns about patient readiness; concerns about patients' mental health and how this may be impacted by discussing a potentially upsetting and stigmatising topic; and lack of effective and individualised treatment and/or referral options. The findings suggest that responses to overweight and obesity need to be localised and tailored. Structural-level change is required to enable better responses to overweight and obesity, including multidisciplinary team approaches.


Predicting Pain and Disability After Shoulder Arthroscopy: Rotator Cuff Tear Severity and Concomitant Arthroscopic Procedures.
Study the influence of concomitant arthroscopic procedures and rotator cuff tear (RCT) severity on daily pain intensity and disability after shoulder arthroscopy. A sample of 145 patients (mean age, 42.66±17.6; number of males=96) completed daily pain intensity and disability measures before shoulder arthroscopy. RCT severity was assessed on a 7-point scale based on operative report. Operative procedures were categorized based on anatomic location and invasiveness to calculate a weighted concomitant procedure estimate. Postoperative daily pain and disability were assessed 3 months, 6 months, and 1 year status after surgery. Only concomitant procedures were associated with 3-month daily pain intensity after adjusting for age, sex, pain duration, and baseline daily pain intensity (R=0.03; standardized β=0.203; P=0.036). Moreover, post hoc analysis revealed concomitant procedures remained predictive of 3-month daily pain intensity when the estimate was calculated independent of RCT procedures (R=0.05; standardized β=0.233; P=0.007), and when accounting for postoperative factors of physical therapy and analgesic medication (R=0.05; standardized β=0.237; P=0.007). Neither RCT severity nor concomitant procedures were predictive of daily pain intensity at 6 months and 1 year, or disability at any time-point, in the adjusted models (P>0.05). This study introduces a novel means for estimating concurrent procedures for use as a prognostic factor. Our findings are similar to previous research of RCT severity that was found to have limited influence on postoperative outcomes. In contrast, this study highlights the potential importance of accounting for concomitant arthroscopic procedures as a predictor of early postoperative pain intensity outcomes.


Modelling the response of FOXO transcription factors to multiple post-translational modifications made by ageing-related signalling pathways.
FOXO transcription factors are an important, conserved family of regulators of cellular processes including metabolism, cell-cycle progression, apoptosis and stress resistance. They are required for the efficacy of several of the genetic interventions that modulate lifespan. FOXO activity is regulated by multiple post-translational modifications (PTMs) that affect its subcellular localization, half-life, DNA binding and transcriptional activity. Here, we show how a mathematical modelling approach can be used to simulate the effects, singly and in combination, of these PTMs. Our model is implemented using the Systems Biology Markup Language (SBML), generated by an ancillary program and simulated in a stochastic framework. The use of the ancillary program to generate the SBML is necessary because the possibility that many regulatory PTMs may be added, each independently of the others, means that a large number of chemically distinct forms of the FOXO molecule must be taken into account, and the program is used to generate them. Although the model does not yet include detailed representations of events upstream and downstream of FOXO, we show how it can qualitatively, and in some cases quantitatively, reproduce the known effects of certain treatments that induce various single and multiple PTMs, and allows for a complex spatiotemporal interplay of effects due to the activation of multiple PTM-inducing treatments. Thus, it provides an important framework to integrate current knowledge about the behaviour of FOXO. The approach should be generally applicable to other proteins experiencing multiple regulations.


Member risk adjustment for ambulatory episodes of care.
The purpose of this study was to model health-plan member risk based on member characteristics in order to separate member risk from other utilization determinants for the use of health care services across sites of care. The approach was to build episodes of care (EOCs) by sorting one year of encounter/claims data into Common Treatment Categories (CTCs). These data came from a variety of health plans, both capitated and non-capitated, covering over 2 million lives. The EOCs were characterized by an array of event and intensity measures. Episode-level risk for each of these measures was modeled by regressions based on member demographic and clinical characteristics. The results of this study show that member characteristics explain a substantial amount of event and intensity variation within episodes and that no single performance measure can summarize the care of health plan members. This method for evaluating member risk can be used both to stratify members according to their future risk and potentially to assess provider or health plan performance or to adjust reimbursement for performance or risk selection.


Characterization of a chimpanzee anti-human melanoma antiserum.
An antiserum to human melanoma has been produced in a chimpanzee by hyperimmunization with melanoma cells from a single donor. After absorption with the peripheral blood lymphocytes of the tumor donor, this antiserum is specifically cytotoxic to melanoma cells from 14 tissue culture cell lines and to cells from 8 fetal fibroblast cell lines. Peripheral blood lymphocytes-absorbed antiserum is negative when tested against a large panel of normal peripheral blood lymphocytes and fibroblasts and against eight non-melanoma neoplastic tissue culture cell lines. Subsequent absorption with melanoma cells from any of seven sources removes all antimelonoma and all antifetal reactivity of the antiserum. Similar absorption with fetal cells removes all antifetal activity but does not completely remove the antimelanoma activity. The immunoglobulin G fraction of the antiserum is shown to possess the cytotoxic activity.


An interval two-stage fuzzy fractional programming model for planning water resources management in the coastal region - A case study of Shenzhen, China.
In this study, an interval two-stage fuzzy fractional programming (TFFP) method is developed to facilitate collaborative governance of economy and water resources. Methods of interval programming, fuzzy programming, two-stage programming, and fractional programming are integrated within a general system optimization framework. The main contribution of TFFP is simultaneously addressing various uncertainties and tackling trade-offs between environmental and economic objectives in the optimized schemes for water resources allocation. A case study of a highly urbanized coastal city (i.e., Shenzhen) in China is provided as an example for demonstrating the proposed approach. According to the results, industrial sectors should receive 34.8% of total water supply, while agricultural sectors should receive 1.5%. For the spatial allocation of water resources, Bao An, Long Gang, and Fu Tian districts should be allocated 21.6%, 20.5%, and 14.8% water to promote the economic development. The discharge analysis indicates that chemical oxygen demand (CODcr) and total phosphorus (TP) would be key pollutants. Moreover, the optimized seawater desalination volume would be negligibly influenced by price, while the upper bounds of desalination would be increased with the raising acceptable credibility levels in the period of 2031-2035. Analysis of desalination prices also reveals that the decision-makers should increase the scale of desalination in the period of 2021-2025. In addition, the effectiveness and applicability of TFFP would be evaluated under economic maximization scenarios. The result showed that the economic maximization scenario could obtain higher economic benefits, but it would be accompanied by a larger number of pollutant discharges. It is expected that this study will provide solid bases for planning water resources management systems in coastal regions.


[Genetic heteroduplex analysis of the R6K plasmid].
The results of genetic studies of R6K Tra1- and R6Kdelta[Sm1] mutants of R6K plasmid and those of heteroduplex analysis of DNAs have shown that DNA of this drug-resistant factor contains three loops flanked by the inverted repeats. The latter are designated as IR1, IR2 and IR3 and are of 50, 100 and 120 nucleotides in size respectively. IR1 is inserted into the loop flanked by IR2. Loops with these two repeats are located in major EcoR1 fragment, IR3 having been found in minor EcoRI fragment of the plasmid. The evidence obtained from the analysis of heteroduplex R6K/RSF2124 has shown that the loop with IR1 is corresponding to transposon Tn3. The extent of the deletion deltaSm1 indicates that IR2 may be a part of a transposon bearing the resistance to streptomycin. By comparing present data with those obtaine from the analysis of the RSF1040 factor of DNA replication initiation sites (Grosa et al., 1976), it has been suggested that the loop with IR3 represents a transposon with replicative functions (TnRep). The deletion of the mutant plasmid R6Kdelta[Sm1] (7.2 . 10(6) daltons in size) which affected one of the EcoRI sites not only confers the sensitivity to streptomycin but enhances also the efficiency of conjugational transfer and results in the loss of the R6K ability to bring about integrative suppression and to inhibit the fertility of the plasmids from IncP and IncN groups. The deletion mutant proved to have lost the property of incompatibility with the initial plasmid R6K and with itself.


Dry matter intake and digestion of alfalfa harvested at sunset and sunrise.
The preference exhibited by animals in selecting one feed over another is important only if the preferred diet is consumed daily in larger quantities, digested to a greater extent, or both. Six alfalfa (Medicago sativa L.) hays were harvested in pairs at sunset (PM) and sunrise (AM) on consecutive days at three harvest dates. A previous study of these hays demonstrated differences in ruminant preference favoring PM harvests. This study evaluated the effects of time of cutting and harvest date on voluntary DMI and nutrient digestibility. The hays were field-cured, baled, and chopped before evaluation for intake and digestibility. Studies were conducted for sheep (Ovis aries), goats (Capra hircus), and cattle (Bos taurus). Goats, but not steers or sheep, demonstrated differences in nutrient digestibility between PM- and AM-cut hays. Goats consumed more PM than AM hay (2.97 vs. 2.83 kg/100 kg of BW; P = 0.07) and digested it to a greater extent (0.710 vs. 0.696; P = 0.03), resulting in greater digestible DMI (2.11 vs. 1.97 kg/100 kg of BW; P = 0.03). Sheep consumed (mean = 2.52 kg/100 kg of BW; P = 0.59) and digested (mean = 0.681; P = 0.25) PM- and AM-cut hays similarly. Steers consumed larger quantities of PM-than AM-cut hay (2.90 vs. 2.62 kg/100 kg of BW; P = 0.11), but digestion did not differ with cutting time (mean = 0.660; P = 0.75). Difference values (composition of fed hay minus composition of orts) indicated that sheep and goats selected from the feed offered similarly, whereas steers selected differently. Difference values for CP averaged 94 and 101 g/kg for goats and sheep and 32 g/kg for steers (P < 0.01), and difference values for NDF averaged 185 and 196 g/kg for goats and sheep and 73 g/kg for steers (P <or= 0.01). Steer DMI and digestible DMI were associated with preference (r = +0.83, P <or= 0.05; and r = +0.89, P <or= 0.05) and with coordinates for preference criteria (dimension 1; r = +0.90, P <or= 0.05; and r = +0.89, P <or= 0.05) from a previous preference trial. Intake and digestion responses for goats and sheep showed no relationship with the previous preference trial measurements. For cattle and goats, the management strategy of mowing in the afternoon seems to take advantage of small, but influential diurnal changes in the soluble carbohydrate fraction and offers the potential to improve forage quality.


Distinct CCR2(+) Gr1(+) cells control growth of the Yersinia pestis ΔyopM mutant in liver and spleen during systemic plague.
We are using a systemic plague model to identify the cells and pathways that are undermined by the virulence protein YopM of the plague bacterium Yersinia pestis. In this study, we pursued previous findings that Gr1(+) cells are required to selectively limit growth of ΔyopM Y. pestis and that CD11b(+) cells other than polymorphonuclear leukocytes (PMNs) are selectively lost in spleens infected with parent Y. pestis. When PMNs were ablated from mice, ΔyopM Y. pestis grew as well as the parent strain in liver but not in spleen, showing that these cells are critical for controlling growth of the mutant in liver but not spleen. In mice lacking expression of the chemokine receptor CCR2, wild-type growth was restored to ΔyopM Y. pestis in both organs. In spleen, the Gr1(+) cells differentially recruited by parent and ΔyopM Y. pestis infections were CCR2(+) Gr1(+) CD11b(+) CD11c(Lo-Int) MAC3(+) iNOS(+) (inducible nitric oxide synthase-positive) inflammatory dendritic cells (iDCs), and their recruitment to spleen from blood was blocked when YopM was present in the infecting strain. Consistent with influx of iDCs being affected by YopM in spleen, the growth defect of the ΔyopM mutant was relieved by the parent Y. pestis strain in a coinfection assay in which the parent strain could affect the fate of the mutant in trans. In a mouse model of bubonic plague, CCR2 also was shown to be required for ΔyopM Y. pestis to show wild-type growth in skin. The data imply that YopM's pathogenic effect indirectly undermines signaling through CCR2. We propose a model for how YopM exerts its different effects in liver and spleen.


Prospective Integrated Individualized Clinical Decision-making and Outcome Evaluation for Surgery in Patients with Acromegaly: A New Paradigm?
Growth-hormone-producing pituitary adenomas have variable likelihood for biochemical remission (BR). During preoperative counseling, individual estimated surgical likelihoods/risks should be balanced against alternative (medical) treatments, which is necessary for accurate outcome presentation. Preoperative estimation of BR or total resection (TR) likelihoods have not been reported, resulting in extrapolation of individual outcomes. To share an innovative outcome reporting paradigm by integrating surgical decision-making, and expected/realized results, resulting from the Value-Based Health Care (VBHC) care path with periodical performance evaluation and care innovation cycle. Prospective cohort study of consecutive patients with acromegaly undergoing surgery (January 2016-December 2020; postoperative follow-up ≥6 months) reporting on both classic, and novel innovative outcome evaluations. Fifty eight patients (66 procedures) were included. Intended TR was achieved in 34/50 procedures, whereas intended debulking was achieved in 15/16 procedures. 38/66 procedures resulted in BR, and 4 procedures resulted in permanent complications. Achieving intended surgical goal was estimated preoperatively as likely in 33 (goal achieved (GA) in 28/33), potentially in 27 (GA in 19/27), and unlikely in 6 procedures (GA in 2/6). Integrated Outcome Square 1 (IOQ1) -intended effect achieved without complications- was achieved in 46/66 patients. Implementation of the developed quality process positively affects preoperative individual shared decision-making, resulting in improved (individual) outcomes, particularly in complex patients for whom preoperative chances are not fully reflected by tumor size and KNOSP grade, e.g., reoperations, or other challenging circumstances identified during preoperative counseling. Through repeated evaluations, our own team's knowledge increased, allowing for improved individualized treatment strategies.


Wide temperature range testing with ROTEM coagulation analyses.
Mild induced hypothermia is used for neuroprotection in patients successfully resuscitated after cardiac arrest. Temperature-dependent effects on rotational thromboelastometry (ROTEM(®)) assays with EXTEM(®), FIBTEM(®), or APTEM(®) in cardiac arrest patients have not previously been studied. Ten patients with out-of-hospital cardiac arrest who underwent induced hypothermia were studied during stable hypothermia at 33°C. ROTEM temperature effects on EXTEM, FIBTEM, and APTEM assays were studied at temperatures set between 30°C and 42°C. Citrated whole blood test tubes were incubated in temperature-adjusted heating blocks and then investigated at respective temperature in the temperature-adjusted ROTEM. The following variables were determined: clotting time (CT), clot formation time (CFT), α-angle, and maximum clot firmness (MCF). The results from hypo- and hyperthermia samples were compared with the samples incubated at 37°C using the Wilcoxon matched-pairs signed-rank test. A p-value of <0.05 was considered significant. CT-EXTEM(®) and CT-APTEM(®) were prolonged by hypothermia at 30°C (p<0.01 for both) and 33°C (p<0.05 for both). Hyperthermia at 42°C shortened CT-EXTEM (p<0.05) and CT-APTEM (p<0.01). CFT-EXTEM(®) and CFT-APTEM(®) were markedly prolonged by hypothermia at 30°C, 33°C, and 35°C (p<0.01 for all except CFT-EXTEM, 35°C [p<0.05]). The α-angle-EXTEM was markedly decreased at 30°C, 33°C, and 35°C (p<0.01) but increased at 40°C (p<0.05) and 42°C (p<0.01); α-angle-APTEM showed similar results. MCF was unchanged at different temperatures for all tests. ROTEM (EXTEM, FIBTEM, and APTEM assays) revealed a hypocoagulative response to in vitro-applied hypothermia in the blood of cardiac arrest patients reflected in the prolonged clot initiation and decreased clot propagation. Hyperthermia showed the opposite effects. Clot firmness was not affected by temperature.


Increases in urinary albumin and beta2-microglobulin are independently associated with blood pressure in the Japanese general population: the Takahata Study.
Essential hypertension is a multifactorial disorder and a risk factor for renal failure and cardiovascular disease. Recently it was hypothesized that subtle acquired renal injury such as renal microvascular and tubulointerstitial damage induces salt-sensitive hypertension. The objective of this study was to examine the relationship between blood pressure and renal abnormalities in the Japanese general population. The participants in this community-based, cross-sectional study were 1,965 subjects over 40 years old, without renal insufficiency and antihypertensive medication. Urine albumin-creatinine ratio (UACR) and beta2-microglobulin-creatinine ratio (UBCR) were measured in single spot urine samples, as markers of renal microvascular and tubulointerstitial damage, respectively. Multiple linear regression analysis showed a significant positive correlation of blood pressure with UACR and UBCR, but not with estimated glomerular filtration rate. In multiple logistic regression analysis, the increases in UACR and UBCR were independently associated with hypertension, after adjustment for possible confounders. Higher levels of UACR (≥ 5.9 mg g(-1)) and UBCR (≥ 145 μg g(-1)) were associated with a significantly higher risk of hypertension, compared with UACR ≤ 5.8 mg g(-1) and UBCR ≤ 84.5 μg g(-1), respectively. Furthermore, there was a positive relationship between urinary sodium excretion and blood pressure in subjects with high UBCR tertile. This study showed that the increases in urinary albumin and beta2-microglobulin were independently associated with blood pressure in a general population. These renal abnormalities may be differentially related to the development of hypertension.


CaV1.2 rather than CaV1.3 is coupled to glucose-stimulated insulin secretion in INS-1 832/13 cells.
In clonal beta-cell lines and islets from different species, a variety of calcium channels are coupled to glucose-stimulated insulin secretion. The aim of this study was to identify the voltage-gated calcium channels that control insulin secretion in insulinoma (INS)-1 832/13 cells. The mRNA level of Ca(V)1.2 exceeded that of Ca(V)1.3 and Ca(V)2.3 two-fold. Insulin secretion, which rose tenfold in response to 16.7 mM glucose, was completely abolished by 5 microM isradipine that blocks Ca(V)1.2 and Ca(V)1.3. Similarly, the increase in intracellular calcium in response to 15 mM glucose was decreased in the presence of 5 microM isradipine, and the frequency of calcium spikes was decreased to the level seen at 2.8 mM glucose. By contrast, inhibition of Ca(V)2.3 with 100 nM SNX-482 did not significantly affect insulin secretion or intracellular calcium. Using RNA interference, Ca(V)1.2 mRNA and protein levels were knocked down by approximately 65% and approximately 34% respectively, which reduced insulin secretion in response to 16.7 mM glucose by 50%. Similar reductions in calcium currents and cell capacitance were seen in standard whole-cell patch-clamp experiments. The remaining secretion of insulin could be reduced to the basal level by 5 microM isradipine. Calcium influx underlying this residual insulin secretion could result from persisting Ca(V)1.2 expression in transfected cells since knock-down of Ca(V)1.3 did not affect glucose-stimulated insulin secretion. In summary, our results suggest that Ca(V)1.2 is critical for insulin secretion in INS-1 832/13 cells.


Monocyte to lymphocyte ratio and hemoglobin level to predict tuberculosis after antiretroviral therapy initiation.
To determine the performance of the baseline monocyte to lymphocyte ratio (MLR), baseline anemia severity and combination of these biomarkers, to predict tuberculosis (TB) incidence in people with HIV (PWH) after antiretroviral therapy (ART) initiation. Multicenter, retrospective cohort study. We utilized the data from study A5175 (Prospective Evaluation of Antiretroviral Therapy in Resource-limited Settings: PEARLS). We assessed the utility of MLR, anemia severity and in combination, for predicting TB in the first year after ART. Cox regression was used to assess associations of MLR and anemia with incident TB. Harrell's C index was used to describe single model discrimination. A total of 1455 participants with a median age of 34 [interquartile range (IQR) 29, 41] were included. Fifty-four participants were diagnosed with TB. The hazard ratio (HR) for incident TB was 1.77 [95% confidence interval (CI) 1.01-3.07]; P = 0.04 for those with MLR ≥0.23. The HR for mild/mod anemia was 3.35 (95% CI 1.78-6.29; P < 0.001) and 18.16 (95% CI 5.17-63.77; P < 0.001) for severe anemia. After combining parameters, there were increases in adjusted HR (aHR) for MLR ≥0.23 to 1.83 (95% CI 1.05-3.18), and degrees of anemia to 3.38 (95% CI 1.80-6.35) for mild/mod anemia and 19.09 (95% CI 5.43-67.12) for severe anemia. MLR and hemoglobin levels which are available in routine HIV care can be used at ART initiation for identifying patients at high risk of developing TB disease to guide diagnostic and management decisions.


Potential Role of Cellular Senescence in Asthma.
Cellular senescence is a complicated process featured by irreversible cell cycle arrest and senescence-associated secreted phenotype (SASP), resulting in accumulation of senescent cells, and low-grade inflammation. Cellular senescence not only occurs during the natural aging of normal cells, but also can be accelerated by various pathological factors. Cumulative studies have shown the role of cellular senescence in the pathogenesis of chronic lung diseases including chronic obstructive pulmonary diseases (COPD) and idiopathic pulmonary fibrosis (IPF) by promoting airway inflammation and airway remodeling. Recently, great interest has been raised in the involvement of cellular senescence in asthma. Limited but valuable data has indicated accelerating cellular senescence in asthma. This review will compile current findings regarding the underlying relationship between cellular senescence and asthma, mainly through discussing the potential mechanisms of cellular senescence in asthma, the impact of senescent cells on the pathobiology of asthma, and the efficiency and feasibility of using anti-aging therapies in asthmatic patients.


The recent expansion in the Australian cocaine market: who are the new users and what are the harms?
Cocaine supplies to and within Australia increased after 2006-07, and there is some evidence that cocaine demand may also have risen. However, the extent, nature and public health implications of any changes in cocaine demand remain unclear. Equally unclear, is whether such changes may have been fuelled by declines in two of Australia's other stimulant markets. We examined general population trends in cocaine use and harmful practices and use of related stimulants between 1998 and 2010, and conducted age-period-cohort analyses using five repeated cross-sections of Australia's National Drug Strategy Household Survey. The results indicate past year cocaine use prevalence has increased significantly since 2004, to its highest point in the past 12 years; 2.1% in 2010. But frequency of cocaine use has not increased. Moreover, most harmful practices (injecting, high-quantity use) have remained stable. Changes in the cocaine market appear related to changes in the Australian methamphetamine and ecstasy markets, including declining purity of ecstasy. For example, the cohorts of people most likely to exhibit recent cocaine use were also most likely to have used ecstasy and methamphetamine (those born from 1976 to 1984). The findings indicate that an increase in cocaine demand does not necessarily lead to substantial increases in public health harm: and indeed that the public health implications from the recent increase are likely to be negligible. Moreover, the findings suggest changes to either ecstasy or methamphetamine supply may lead to more shifts in demand for Australia's cocaine market.