Simple, Hackable, Size-Selective, Amine-Functionalized Fe-Oxide Nanoparticles for Biomedical Applications.
A facile one-pot method for synthesizing amine-functionalized nonspherical Fe3O4 nanoparticles in gram-scale quantities is presented using just a single source of iron (iron(II) chloride) and an amine (triethylamine). The amine not only transforms iron salt to Fe3O4, but also directs the morphology of the nanoparticles along with the temperature of the reaction and functionalizes them, making the synthesis very economical. By modifying the surface further, these nanoparticles promise to offer useful biomedical applications. For example, after biocide coating, the particles are found to be 100% effective in deactivating methicillin-resistant Staphylococcus aureus (MRSA) bacteria in 2 h. Cellular-uptake studies using biocompatible EDTA-Na3 (N-(trimethoxysilyl-propyl)ethylenediaminetriacetate, trisodium salt)-coated nanoparticles in human glioblastoma U-251 cells show that the majority of the particles are internalized by the cells in the presence of a small dc-magnetic field, making these particles a potential candidate as drug carriers for magnetic field-targeted delivery and hyperthermia.


A flexible reporter system for direct observation and isolation of cancer stem cells.
Many tumors are hierarchically organized with a minority cell population that has stem-like properties and enhanced ability to initiate tumorigenesis and drive therapeutic relapse. These cancer stem cells (CSCs) are typically identified by complex combinations of cell-surface markers that differ among tumor types. Here, we developed a flexible lentiviral-based reporter system that allows direct visualization of CSCs based on functional properties. The reporter responds to the core stem cell transcription factors OCT4 and SOX2, with further selectivity and kinetic resolution coming from use of a proteasome-targeting degron. Cancer cells marked by this reporter have the expected properties of self-renewal, generation of heterogeneous offspring, high tumor- and metastasis-initiating activity, and resistance to chemotherapeutics. With this approach, the spatial distribution of CSCs can be assessed in settings that retain microenvironmental and structural cues, and CSC plasticity and response to therapeutics can be monitored in real time.


A randomized controlled clinical study of the effect of daily intake of Ascophyllum nodosum alga on calculus, plaque, and gingivitis.
The aim of this study is to evaluate, in a randomized controlled cross-over study, the effect of daily intake of the alga Ascophyllum nodosum on supragingival calculus, plaque formation, and gingival health over a 6-month period. Sixty-one adults with moderate to heavy calculus formation since their last yearly recall visit participated. In a randomized order over two 6-month periods, they swallowed two capsules daily, comprising a total of 500 mg dried marine alga powder (Ascophyllum nodosum, ProDen PlaqueOff®) or two negative control tablets. During the study, the participants maintained their regular oral habits. Their teeth were professionally cleaned at the start of each period and after the 6-month registrations. A wash out period of 1 month separated the two 6-month periods. Supragingival calculus (Volpe Manhold), gingivitis (Löe and Silness), gingival bleeding (Ainamo and Bay), and plaque (Quigley-Hein) were registered at screening and at the end of the two periods. Differences in oral health between the test and control periods were analyzed using a paired t test and Wilcoxon signed rank test. Fifty-five participants completed the study. After the alga intake, the mean calculus reduction was 52% compared to the control (p < 0.0001). Fifty-two participants showed less calculus formation in the alga group than in the control group. Plaque (p = 0.008) and gingival bleeding (p = 0.02) were also significantly less in the alga group. However, no significant difference was found between the groups for gingivitis (p = 0.13). The alga intake significantly reduced the formation of supragingival calculus and plaque and occurrence of gingival bleeding. The alga has a systemic effect on oral health. Daily intake of the alga Ascophyllum nodosum as an adjunct to customary oral hygiene showed a major reduction of supragingival calculus formation and reduced plaque formation. In addition, the calculus in the alga group was characterized by a more porous and less solid structure and was easier to remove than the calculus in the control group.


Vascular complications after percutaneous coronary intervention following hemostasis with the Mynx vascular closure device versus the AngioSeal vascular closure device.
We investigated the prevalence of vascular complications after PCI following hemostasis in 190 patients (67% men and 33% women, mean age 64 years) treated with the AngioSeal vascular closure device (St. Jude Medical, Austin, Texas) versus 238 patients (67% men and 33% women, mean age 64 years) treated with the Mynx vascular closure device (AccessClosure, Mountain View, California). Death, myocardial infarction or stroke occurred in none of the 190 patients (0%) treated with the AngioSeal versus none of 238 patients (0%) treated with the Mynx. Major vascular complications occurred in 4 of 190 patients (2.1%) treated with the AngioSeal versus 5 of 238 patients (2.1%) treated with the Mynx (p not significant). Major vascular complications in patients treated with the AngioSeal included removal of a malfunctioning device (1.1%), hemorrhage requiring intervention (0.5%) and hemorrhage with a loss of > 3g Hgb (0.5%). The major vascular complications in patients treated with the Mynx included retroperitoneal bleeding requiring surgical intervention (0.8%), pseudoaneurysm with surgical repair (0.8%) and hemorrhage with a loss of > 3g Hgb (0.4%). These complications were not significantly different between the two vascular closure devices (p = 0.77). Minor complications included hematoma > 5 cm (0.5%, n = 1) within the AngioSeal group, as well as procedure failure requiring > 30 minutes of manual compression after device deployment, which occurred in 7 out of 190 patients (3.7%) treated with the AngioSeal versus 22 of 238 patients with the Mynx (9.2%) (p = 0.033). Major vascular complications after PCI following hemostasis with vascular closure devices occurred in 2.1% of 190 patients treated with the AngioSeal vascular closure device versus 2.1% of 238 patients treated with the Mynx vascular closure device (p not significant). The Mynx vascular closure device appears to have a higher rate of device failure.


Bayesian Gaussian processes for identifying the deteriorating patient.
The step-down unit (SDU) is a high-acuity hospital environment, to which patients may be sent after discharge from the intensive care unit (ICU). About 1- in-7 patients will deteriorate in the SDU and require emergency readmission to the ICU. Upon readmission, these patients experience significantly higher mortality risks and lengths of stay. Gaussian process regression (GPR) models are proposed as a flexible, principled, probabilistic method to address the clinical need to monitor continuously patient time-series of vital signs acquired in the SDU. The proposed GPR models focus on the robust forecasting of patient heart rate time-series and on the early detection of patient deterioration. The proposed methods are tested with an SDU data set from the University of Pittsburgh Medical Center, comprising 333 patients, 59 of whom had at least one verified clinical emergency event. Results suggest that GPR-based heart rate monitoring provides superior advanced warning of deterioration compared to the current clinical practice of rules-based thresholding, and slightly outperforms the current state-of-the-art kernel density method, which requires 4 additional vital sign features.


Etiologic classification of TIA and minor stroke by A-S-C-O and causative classification system as compared to TOAST reduces the proportion of patients categorized as cause undetermined.
The assortment of patients based on the underlying pathophysiology is central to preventing recurrent stroke after a transient ischemic attack and minor stroke (TIA-MS). The causative classification of stroke (CCS) and the A-S-C-O (A for atherosclerosis, S for small vessel disease, C for Cardiac source, O for other cause) classification schemes have recently been developed. These systems have not been specifically applied to the TIA-MS population. We hypothesized that both CCS and A-S-C-O would increase the proportion of patients with a definitive etiologic mechanism for TIA-MS as compared with TOAST. Patients were analyzed from the CATCH study. A single-stroke physician assigned all patients to an etiologic subtype using published algorithms for TOAST, CCS and ASCO. We compared the proportions in the various categories for each classification scheme and then the association with stroke progression or recurrence was assessed. TOAST, CCS and A-S-C-O classification schemes were applied in 469 TIA-MS patients. When compared to TOAST both CCS (58.0 vs. 65.3%; p < 0.0001) and ASCO grade 1 or 2 (37.5 vs. 65.3%; p < 0.0001) assigned fewer patients as cause undetermined. CCS had increased assignment of cardioembolism (+3.8%, p = 0.0001) as compared with TOAST. ASCO grade 1 or 2 had increased assignment of cardioembolism (+8.5%, p < 0.0001), large artery atherosclerosis (+14.9%, p < 0.0001) and small artery occlusion (+4.3%, p < 0.0001) as compared with TOAST. Compared with CCS, using ASCO resulted in a 20.5% absolute reduction in patients assigned to the 'cause undetermined' category (p < 0.0001). Patients who had multiple high-risk etiologies either by CCS or ASCO classification or an ASCO undetermined classification had a higher chance of having a recurrent event. Both CCS and ASCO schemes reduce the proportion of TIA and minor stroke patients classified as 'cause undetermined.' ASCO resulted in the fewest patients classified as cause undetermined. Stroke recurrence after TIA-MS is highest in patients with multiple high-risk etiologies or cryptogenic stroke classified by ASCO.


Splenectomy Alters Distribution and Turnover but not Numbers or Protective Capacity of de novo Generated Memory CD8 T-Cells.
The spleen is a highly compartmentalized lymphoid organ that allows for efficient antigen presentation and activation of immune responses. Additionally, the spleen itself functions to remove senescent red blood cells, filter bacteria, and sequester platelets. Splenectomy, commonly performed after blunt force trauma or splenomegaly, has been shown to increase risk of certain bacterial and parasitic infections years after removal of the spleen. Although previous studies report defects in memory B-cells and IgM titers in splenectomized patients, the effect of splenectomy on CD8 T-cell responses and memory CD8 T-cell function remains ill defined. Using TCR-transgenic P14 cells, we demonstrate that homeostatic proliferation and representation of pathogen-specific memory CD8 T-cells in the blood are enhanced in splenectomized compared to sham surgery mice. Surprisingly, despite the enhanced turnover, splx mice displayed no changes in total memory CD8 T-cell numbers nor impaired protection against lethal dose challenge with Listeria monocytogenes. Thus, our data suggest that memory CD8 T-cell maintenance and function remain intact in the absence of the spleen.


The radiation-tolerant x-ray monitor.
A vacuum photoelectric detector (monitor) (VPD) designed for plasma tomography, megnetohydrodynamics monitoring, and imaging with the help of thermal x-ray radiation on the ITER facility is described. Laboratory experiments demonstrate that VPD has high sensitivity to thermal x rays and low sensitivity to hard gamma rays and neutrons. The results of tests of a prototype of this monitor on a (60)Co source of gamma rays, on nuclear reactor and its calibration using radiation from an x-ray tube, and tests of its serviceability on the T-10 facility are presented.


Distinct profile of T cell activation in HIV type 2 compared to HIV type 1 infection: differential mechanism for immunoprotection.
The mechanism for the lower rate of disease progression in HIV-2 infection remains undefined. We evaluated T cell activation in a cohort of HIV-infected commercial sex workers in Dakar, Senegal. CD8+ T cell activation was significantly lower in HIV-2- compared to HIV-1-infected volunteers and both groups displayed higher activation levels compared to seronegative individuals. In contrast, CD4+ T cell activation was similar between the HIV-1 and HIV-2 groups and significantly higher compared to the seronegative group. Interestingly, HIV-2-positive volunteers with evidence of Gag-specific CD8+ T cell responses displayed lower CD4+ T cell activation. Our data suggest that the distinct T cell activation profile in HIV-2-positive individuals may reflect on the presence of effective host immune responses in HIV-2 infection.


Determinants of hand dermatitis, urticaria and loss of skin barrier function in professional cleaners in New Zealand.
This study assessed the risk of dermatitis, urticaria and loss of skin barrier function in 425 cleaners and 281 reference workers (retail workers and bus drivers). Symptoms, atopy and skin barrier function were assessed by questionnaire, skin prick tests, and measurement of transepidermal water loss. Cleaners had an increased risk of current (past 3 months) hand/arm dermatitis (14.8% vs. 10.0%; OR = 1.9, p < 0.05) and urticaria (11% vs. 5.3%; OR = 2.4, p < 0.05) and were more likely to have dermatitis as adults (17.6% vs. 11.4%; OR = 1.8, p < 0.05). The risk of atopy was not increased, but associations with symptoms were more pronounced in atopics. Transepidermal water loss was significantly higher in cleaners. Wet-work was a significant risk factor for dermatitis and hand washing and drying significantly reduced the risk of urticaria. In conclusion, cleaners have an increased risk of hand/arm dermatitis, urticaria and loss of skin barrier function.


CpG island methylator phenotype identifies high risk patients among microsatellite stable BRAF mutated colorectal cancers.
The prognostic value of CpG island methylator phenotype (CIMP) in colorectal cancer remains unsettled. We aimed to assess the prognostic value of this phenotype analyzing a total of 1126 tumor samples obtained from two Norwegian consecutive colorectal cancer series. CIMP status was determined by analyzing the 5-markers CAGNA1G, IGF2, NEUROG1, RUNX3 and SOCS1 by quantitative methylation specific PCR (qMSP). The effect of CIMP on time to recurrence (TTR) and overall survival (OS) were determined by uni- and multivariate analyses. Subgroup analyses were conducted according to MSI and BRAF mutation status, disease stage, and also age at time of diagnosis (<60, 60-74, ≥75 years). Patients with CIMP positive tumors demonstrated significantly shorter TTR and worse OS compared to those with CIMP negative tumors (multivariate hazard ratio [95% CI] 1.86 [1.31-2.63] and 1.89 [1.34-2.65], respectively). In stratified analyses, CIMP tumors showed significantly worse outcome among patients with microsatellite stable (MSS, P < 0.001), and MSS BRAF mutated tumors (P < 0.001), a finding that persisted in patients with stage II, III or IV disease, and that remained significant in multivariate analysis (P < 0.01). Consistent results were found for all three age groups. To conclude, CIMP is significantly associated with inferior outcome for colorectal cancer patients, and can stratify the poor prognostic patients with MSS BRAF mutated tumors.


Long-lived Ag10 6+ luminescence and a split DNA scaffold.
Molecular silver clusters emit across the visible to near-infrared, and specific chromophores can be formed using DNA strands. We study C4AC4TC3G that selectively coordinates and encapsulates Ag10 6+, and this chromophore has two distinct electronic transitions. The green emission is strong and prompt with ϕ = 18% and τ = 1.25 ns, and the near-infrared luminescence is weaker, slower with τ = 50 µs, and is partly quenched by oxygen, suggesting phosphorescence. This lifetime can be modulated by the DNA host, and we consider two derivatives of C4AC4TC3G with similar sequences but distinct structures. In one variant, thymine was excised to create an abasic gap in an otherwise intact strand. In the other, the covalent phosphate linkage was removed to split the DNA scaffold into two fragments. In relation to the contiguous strands, the broken template speeds the luminescence decay by twofold, and this difference may be due to greater DNA flexibility. These modifications suggest that a DNA can be structurally tuned to modulate metastable electronic states in its silver cluster adducts.


Diastereoselective osmium-catalyzed vicinal oxyamination of acyclic allylic alcohol derivatives.
The osmium-catalyzed oxyamination of chiral acyclic allylic alcohol derivatives bearing mono- and 1,1-di-substituted double bonds with benzyl N-(4-tosyloxy)carbamate proceeds with high regioselectivity and moderate levels of diastereoselectivity favoring the anti product. The observed stereoselectivity shows a clear and systematic trend with anti:syn ratios increasing in line with the size of substituent at both the allylic stereocenter and double bond α-carbon. The stereoinduction is in accord with the sense of diastereoselectivity predicted by Kishi's empirical rule and a previously reported transition state model for the osmium-catalyzed dihydroxylation of allylic alcohol derivatives. In contrast, allylic alcohol derivatives bearing trisubstituted double bonds show low or no reactivity in the oxyamination reaction affording the syn product in low yield in the cases examined.


Nurse honours.
A trailblazer who set up England's first nurse-led sickle cell intervention and screening service and campaigned for a Mary Seacole memorial was made a dame by the Queen last week.


Fully kinetic particle-in-cell simulations of a deuterium gas puff z pinch.
We present the first fully kinetic, collisional, and electromagnetic simulations of the complete time evolution of a deuterium gas puff z pinch. Recent experiments with 15-MA current pinches have suggested that the dominant neutron-production mechanism is thermonuclear. We observe distinct differences between the kinetic and magnetohydrodynamic simulations in the pinch evolution with the kinetic simulations producing both thermonuclear and beam-target neutrons. The kinetic approach demonstrated in this Letter represents a viable alternative for performing future plasma physics calculations.


Predicting subsequent decline in kidney allograft function from early surveillance biopsies.
Identifying factors that are predictive of allograft loss might be an important step toward prolonging kidney allograft survival. In this study we sought to determine the association between histologic changes on 1-year surveillance biopsies, changes in graft function and survival. This analysis included 292 adults, recipients of kidneys from living donors (69%) or deceased donors (31%), transplanted between 1998 and 2001 and followed up for 46 +/- 14 months. The primary end point was death-censored graft loss or a >50% reduction in GFR beyond 1 year. One-year biopsies were classified as: (i) Normal (N = 87, 30%), (ii) inflammation (N = 6, 2%), (iii) fibrosis (N = 131, 45%), (iv) fibrosis and inflammation (N = 53, 18%) and (v) transplant glomerulopathy (N = 15, 5%). By multivariate Cox analysis, survival related to biopsy classification (HR = 4.2, p = 0.001), graft function (HR = 0.97, p = 0.001) and HLA mismatches (HR = 1.003, p = 0.004). Using normal histology as a reference, fibrosis and inflammation (HR = 8.5, p < 0.0001) and glomerulopathy (HR = 10, p < 0.0001) related to poorer survival but mild fibrosis alone did not. Importantly, the degree of inflammation associated with fibrosis generally did not qualify for the diagnosis of borderline rejection. In conclusion, inflammation and glomerulopathy 1 year post-transplant predict loss of graft function and graft failure independently of function and other variables.


Analysis of the relations between allergen specific LgG antibody and allergic dermatosis of 14 kinds foods.
To use food-specific IgG antibody detection to explore its application in the allergy dermatoses. 181 patients were included from January 2014 to September 2014. Fourteen food-specific IgG antibodies were detected by ELISA. The positive rates of IgG antibody of the patient group and the healthy group were significantly different. The positive rates of IgG antibody of egg, milk, shrimp and crab took a large proportion in three groups of patients with three kinds of allergy dermatoses of urticaria, eczema and allergic dermatitis, the proportion of which was respectively 70.2%, 77.8% and 71.7%. There was mild and moderate intolerance of food in the allergic dermatitis group while there was no distribution difference of food intolerance in urticaria group and eczema group. Among urticaria and allergic dermatitis patients with positive antibody, the positive rate of children was significantly higher than that of adults while there was no significant difference between children and adults among eczema patients with positive antibody. Allergy dermatoses are closely related to food-specific IgG antibody and the allergy dermatoses patients have a high incidence rate of food intolerance; detecting IgG antibody in patients is of great significance for the diagnosis and treatment of allergy dermatoses.


An overview of colorectal cancer screening.
Colorectal cancer ranks highly amongst all cancer sites in incidence and contributes to a substantial number of cancer related deaths in the United Kingdom. However, screening of average risk individuals has been shown to reduce both disease associated mortality and incidence. This paper provides an overview of both current and future screening methods for colorectal cancer, as well as current practice for screening in both average and high risk individuals.


Antiphospholipid antibodies and the protein C pathway.
Haemostasis is a delicate balance between procoagulant and anticoagulant processes. In the human body usually anticoagulant mechanisms prevail over procoagulant mechanisms, thereby preventing a prothrombotic state. The antiphospholipid syndrome is an example in which this balance is shifted to a more prothrombotic state due to the presence of antiphospholipid antibodies. One of the most extensively proposed pathogenic mechanisms within the antiphospholipid syndrome is the inhibition of protein C by antiphospholipid antibodies. Antiphospholipid antibodies have been described to have different actions on the protein C pathway, for example decreasing protein C and/or S plasma levels, inducing increased resistance against activated protein C and lowering thrombin levels (resulting in an impaired protein C activation). This review briefly discusses the actions of protein C in human body but mainly focuses on the effects of antiphospholipid antibodies on the protein C pathway that have been described in literature.


Vero cells injected with adenovirus type 2 mRNA produce authentic viral polypeptide patterns: early mRNA promotes growth of adenovirus-associated virus.
Adenovirus type 2 mRNAs were injected via glass capillaries into Vero cells, a line of African green monkey kidney cells permissive for adenovirus growth. Polypeptides synthesized after injection were labeled with 35S-labeled amino acids, precipitated with antiviral sera, and analyzed by polyacrylamide gel electrophoresis. Both early and late viral mRNAs give rise to authentic polypeptides. The in vivo translation of mRNAs can be measured as late as 24 hr after injection. The ability to analyze the protein products of microinjected mRNAs directly should greatly extend the applications of the procedure. Vero cells injected with early mRNA from adenovirus type 2 support the growth of adenovirus-associated virus, a defective virus that is dependent on adenovirus helper functions. This result demonstrates that a measurable biological activity, the ability to overcome the defectiveness of adenovirus-associated virus, resides in early adenovirus mRNA.


Electrospun oriented gelatin-hydroxyapatite fiber scaffolds for bone tissue engineering.
Tissue engineering of human fetal osteoblast cells was investigated on gelatin-hydroxyapatite (HA), crosslinked, electrospun oriented fiber scaffolds at the different HA concentrations of 0, 10, 20, and 25 wt % in the dry fibers and different fiber diameter, pore size and porosity of scaffolds. Rheological tests and proton nuclear magnetic resonance spectroscopy were conducted for all solutions used for electrospinning. It was found that 25 wt % HA-gelatin scaffolds electrospun at 20 kV led to the greatest cell attachment, cell proliferation and extracellular matrix (ECM) production while fiber orientation improved the mechanical properties, where crosslinked electrospun 25 wt % HA-gelatin fiber scaffolds yielded a Young's modulus in the range of 0.5-0.9 GPa and a tensile strength in the range of 4-10 MPa in the fiber direction for an applied voltage of 20-30 kV, respectively, in the electrospinning of scaffolds. Biological characterization of cell seeded scaffolds yielded the rate of cell growth and ECM (collagen and calcium) production by the cells as a function of time; it included cell seeding efficiency tests, alamar blue cell proliferation assay, alkaline phosphate (ALP) assay, collagen assay, calcium colorimetric assay, fluorescence microscopy for live and dead cells, and scanning electron microscopy for cell culture from 1 to 18 days. After 18 days, cells seeded and grown on the 25 wt % HA-gelatin scaffold, electrospun at 20 kV, reached production of collagen at 370 μg/L and calcium production at 0.8 mM. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1911-1926, 2017.


Undernutrition reduces kisspeptin and neurokinin B expression in castrated male sheep.
Undernutrition impairs reproductive success through suppression of gonadotropin-releasing hormone (GnRH), and subsequently luteinizing hormone (LH), secretion. Given that kisspeptin and neurokinin B (NKB) neurons in the arcuate nucleus (ARC) of the hypothalamus are thought to play key stimulatory roles in the generation of GnRH/LH pulses, we hypothesized that feed restriction would reduce the ARC mRNA abundance and protein expression of kisspeptin and NKB in young, male sheep. Fourteen wethers (castrated male sheep five months of age) were either fed to maintain (FM; n = 6) pre-study body weight or feed-restricted (FR; n = 8) to lose 20% of pre-study body weight over 13 weeks. Throughout the study, weekly blood samples were collected and assessed for LH concentration using RIA. At Week 13 of the experiment, animals were killed, heads were perfused with 4% paraformaldehyde, and brain tissue containing the hypothalamus was collected, sectioned, and processed for detection of mRNA (RNAscope) and protein (immunohistochemistry) for kisspeptin and NKB. Mean LH was significantly lower and LH inter-pulse interval was significantly higher in FR wethers compared to FM wethers at the end of the experiment (Week 13). RNAscope analysis revealed significantly fewer cells expressing mRNA for kisspeptin and NKB in FR wethers compared to FM controls, and immunohistochemical analysis revealed significantly fewer immunopositive kisspeptin and NKB cells in FR wethers compared to FM wethers. Taken together, this data supports the idea that long-term feed restriction regulates GnRH/LH secretion through central suppression of kisspeptin and NKB in male sheep. While undernutrition is known to impair reproduction at the level of the brain, the components responsible for this in the brain remain to be fully understood. Using male sheep we examined the effect of undernutrition on two stimulatory molecules in the brain critical for reproduction: kisspeptin and neurokinin B. Feed restriction for several weeks resulted in decreased luteinizing hormone in the blood indicating reproductive function was suppressed. In addition, undernutrition also reduced both kisspeptin and neurokinin B levels within a region of the brain involved in reproduction, the hypothalamus. Given that they have stimulatory roles in reproduction, we believe that undernutrition acts in the brain to reduce kisspeptin and neurokinin B levels leading to the reduction in luteinizing hormone secretion. In summary, long-term undernutrition inhibits reproductive function in sheep through suppression of kisspeptin and neurokinin B within the brain.


Interaction between gfp-tagged Pseudomonas tolaasii P12 and Pleurotus eryngii.
Pseudomonas sp., (formerly reported as strain P12) which produces brown blotch disease symptoms on Pleurotus eryngii, has been identified as P. tolaasii based on its biochemical, physiological properties and 16S rDNA sequence analysis. This pathogen is able to infect basidiocarps when surface-inoculated on mushroom casing soil. However, infected basidiocarps develop the brown blotch disease symptoms when the pathogen concentration in the fruiting body tissues is higher than 10(4) cfu/g d.w. Using gfp-tagged cells and confocal laser scanning microscopy, it was possible to show that the pathogen has the ability to tightly attach to the hyphae of Pleurotus eryngii.


Should an order to "monitor long-term vital signs," including mortality outcomes, be as routine as an order for a laboratory test in patients with rheumatic diseases?
Vital signs alert a health professional to problems which may threaten patient well-being and survival. Clinicians are highly familiar with vital signs for acute disease such as blood pressure, pulse, temperature, but unfamiliar with vital signs for chronic disease, such as physical function, pain, global status, exercise frequency and smoking. Long-term vital signs should be collected at each visit and stored in a computer database, ideally in a flow sheet format, as the memory of clinicians and patients is not reliable over long periods. The structure of the database should be identical from one site to another, so that data may be pooled to analyze large series of patients, particularly those with rare diseases, such as systemic sclerosis, polymyositis, and vasculitis. Of course, appropriate additional information beyond simple "long-term vital signs" from a physical examination, radiograph and laboratory are needed for further accurate assessment of prognosis and outcomes, in both acute and chronic diseases, and optimal information will emerge from specialized research centers. A common long-term vital signs database would be a major advance from current descriptive, non-quantitative monitoring of patients with rheumatic diseases, and would allow any rheumatologist to contribute to improved knowledge and mortality outcomes of rheumatic diseases.


Preschool Programs that Help Families Promote Child Social-Emotional School Readiness: Promising New Strategies.
Parents play a central role in supporting the early learning that positions young children for success when they enter formal schooling. For this reason, efforts to engage families in meaningful collaboration is a long-standing goal of high-quality early childhood education (ECE). Family-school engagement can take multiple forms; in this review, we focus on universal preschool-based outreach strategies that help parents support growth in child social-emotional and self-regulation competencies and prepare them for the transition into formal schooling. Recent research has expanded understanding of the neurodevelopmental processes that underlie child school readiness, and the impact of parenting (and the social ecology affecting parenting) on those processes. These new insights have fueled innovation in preschool-based efforts to partner with and support parents, expanding and shifting the focus of that programming. In addition, new approaches to intervention design and delivery are emerging to address the pervasive challenges of reaching and engaging families, especially those representing diverse racial, ethnic, cultural, and socioeconomic backgrounds. This paper reviews developmental research that underscores the importance of prioritizing child social-emotional learning (with attention to self-regulation and approaches to learning) in universal preschool-based parenting programs targeting young children. We highlight the intervention strategies used in programs with strong evidence of impact on child readiness and school adjustment based on randomized controlled trials (RCTs). New directions in intervention design and delivery strategies are highlighted, with the hope of extending intervention reach and improving family engagement and benefit.


The chloroplast genome of Phacus orbicularis (Euglenophyceae): an initial datum point for the phacaceae.
The Euglenophyceae chloroplast was acquired when a heterotrophic euglenoid engulfed a green alga and subsequently retained the algal chloroplast, in a process known as secondary endosymbiosis. Since this event, Euglenophyceae have diverged widely and their chloroplast genomes (cpGenomes) have as well. Changes to the cpGenome include extensive gene rearrangement and the proliferation of introns, the analyses of which have proven to be useful in examining cpGenome changes throughout the Euglenophyceae. The Euglenales fall into two families, Euglenaceae and Phacaceae. Euglenaceae contains eight genera and at least one cpGenome has been published for each genus. Phacaceae, on the other hand, contains three genera, none of which have had a representative chloroplast genome sequenced. Members of this family have many small disk-shaped chloroplasts that lack pyrenoids. We sequenced and annotated the cpGenome of Phacus orbicularis in order to fill in the large gap in our understanding of Euglenophyceae cpGenome evolution, especially in regard to intron number and gene order. We compared this cpGenome to those of species from both the Euglenaceae and Eutreptiales of the Euglenophyceae phylogenetic tree. The cpGenome showed characteristics that were more derived than that of the basal species Eutreptia viridis, with extensive gene rearrangements and nearly three times as many introns. In contrast, it contained fewer introns than all but one of the previously reported Euglenaceae cpGenomes, had a smaller estimated genome size, and shared greater synteny with two main branches of that family.


Identification and characterization of anonymous nuclear markers for the double-striped cockroach, Blattella bisignata.
During the last decade, multilocus analysis has gradually become a powerful tool for the studies of population genetics and phylogeography. The double-striped cockroach, Blattella bisignata, is endemic to southeast Asia, and there is currently little genetic information available for the species. We chose it as the target species to investigate a biodiversity hotspot in southwest China. Here, we report the identification and characterization of 11 single-copy anonymous nuclear markers with an average length of 378bp. These loci, isolated from a genomic library of B. bisignata, can amplify in two additional Blattella species (B. germanica and B. lituricollis). While testing these markers in representative species of Blattellidae, Blattidae and Epilampridae, some of them can cross-amplify successfully. After sequencing 30 individuals collected from southern China per locus, we found relatively high variability (approximately 3.6 SNPs per 100bp). Finally, a small-scale study was also performed to show that these markers do indeed fulfill the expectations as phylogeographic markers.


Inhibition of CD26/dipeptidyl peptidase IV enhances CCL11/eotaxin-mediated recruitment of eosinophils in vivo.
Chemokines mediate the recruitment of leukocytes to the sites of inflammation. N-terminal truncation of chemokines by the protease dipeptidyl peptidase IV (DPPIV) potentially restricts their activity during inflammatory processes such as allergic reactions, but direct evidence in vivo is very rare. After demonstrating that N-terminal truncation of the chemokine CCL11/eotaxin by DPPIV results in a loss of CCR3-mediated intracellular calcium mobilization and CCR3 internalization in human eosinophils, we focused on the in vivo role of CCL11 and provide direct evidence for specific kinetic and rate-determining effects by DPPIV-like enzymatic activity on CCL11-mediated responses of eosinophils. Namely, it is demonstrated that i.v. administration of CCL11 in wild-type F344 rats leads to mobilization of eosinophils into the blood, peaking at 30 min. This mobilization is significantly increased in DPPIV-deficient F344 rats. Intradermal administration of CCL11 is followed by a dose-dependent recruitment of eosinophils into the skin and is significantly more effective in DPPIV-deficient F344 mutants as well as after pharmacological inhibition of DPPIV. Interestingly, CCL11 application leads to an up-regulation of DPPIV, which is not associated with negative feedback inhibition via DPPIV-cleaved CCL11((3-74)). These findings demonstrate regulatory effects of DPPIV for the recruitment of eosinophils. Furthermore, they illustrate that inhibitors of DPPIV have the potential to interfere with chemokine-mediated effects in vivo including but not limited to allergy.


Solvent/additive-free synthesis of porous/zeolitic metal azolate frameworks from metal oxide/hydroxide.
Porous/zeolitic metal azolate frameworks may be efficiently prepared, with water as the only byproduct, by heating a mixture of metal oxide/hydroxide and azole ligand.


Activation of antioxidant response element (ARE)-dependent genes by roasted coffee extracts.
Coffee beans contain numerous bioactive components that exhibit antioxidant capacity when assessed using both chemical, cell free, and biological, cell-based model systems. However, the mechanisms underlying the antioxidant effects of coffee in biological systems are not totally understood and in some cases vary considerably from results obtained with simpler in vitro chemical assays. In the present study, the physicochemical characteristics and antioxidant activity of roasted and non-roasted coffee extracts were investigated in both cell free (ORAC(FL)) and cell-based systems. A profile of antioxidant gene expression in cultured human colon adenocarcinoma Caco-2 cells treated with both roasted and non-roasted coffee extracts, respectively, was investigated using Real-Time polymerase chain reaction (PCR) array technology. Results demonstrated that the mechanisms of the antioxidant activity associated with coffee constituents assessed by the ORAC(FL) assay were different to those observed using an intracellular oxidation assay with Caco-2 cells. Moreover, roasted coffee (both light and dark roasted) extracts produced both increased- and decreased-expressions of numerous genes that are involved in the management of oxidative stress via the antioxidant defence system. The selective and specific positive induction of antioxidant response element (ARE)-dependent genes, including gastrointestinal glutathione peroxidase (GPX2), sulfiredoxin (SRXN1), thioredoxin reductase 1 (TXNRD1), peroxiredoxin 1 (PRDX1), peroxiredoxin 4 (PDRX4) and peroxiredoxin 6 (PDRX6) were identified with the activation of the endogenous antioxidant defence system in Caco-2 cells.


Is the association between health-related quality of life and fatigue mediated by depression in patients with multiple sclerosis? A Spanish cross-sectional study.
To determine the mediating effects of depression on health-related quality of life and fatigue in individuals with multiple sclerosis (MS). A cross-sectional study. Tertiary urban hospital. One hundred and eight patients (54% women) with MS participated in this study. Demographic and clinical data (weight, height, medication and neurological impairment), fatigue (Fatigue Impact Scale), depression (Beck Depression Inventory-II) and health-related quality of life (Short-Form Health Survey 36) were collected. Fatigue was significantly associated with bodily pain, physical function, mental health and depression. Depression was associated with bodily pain and mental health. The path analysis found direct effects from physical function, bodily pain and depression to fatigue (all, P<0.01). The path model analysis revealed that depression exerted a mediator effect from bodily pain to fatigue (B=-0.04, P<0.01), and from mental health to fatigue (B=-0.16, P<0.01). The amount of fatigue explained by all predictors in the path model was 37%. This study found that depression mediates the relationship between some health-related quality of life domains and fatigue in people with MS. Future longitudinal studies focusing on proper management of depressive symptoms in individuals with MS will help determine the clinical implications of these findings.


Preoperative evaluation of neurovascular relationship in trigeminal neuralgia by three-dimensional fast low angle shot (3D-FLASH) and three-dimensional constructive interference in steady-state (3D-CISS) MRI sequence.
The purpose of the study was to evaluate the value of high-resolution three-dimensional fast low angle shot (3D-FLASH) and three-dimensional constructive interference in steady-state (3D-CISS) MRI sequence solely or the combination of both in the visualization of neurovascular relationship in patients with trigeminal neuralgia (TN). 65 patients with unilateral TN underwent 3D-FLASH and 3D-CISS imaging were retrospectively studied. Neurovascular relationship at the intracisternal segment of trigeminal nerve was reviewed by two experienced neuroradiologist, who was blinded to the clinical details. The imaging results were compared with the operative findings in all patients. The accuracy and positive rates of the 3D-FLASH + CISS imaging (98.46, 92.31%) in judging the symptomatic side according to the presence of vascular contacts were higher than those of 3D-CISS (90.77%, 84.62) or 3D-FLASH (89.23, 83.08%) sequence. In addition, the statistical analysis showed the sensitivity and accuracy of 3D-FLASH + CISS imaging was higher than that of 3D-FLASH (p < 0.05). The 3D-FLASH + CISS imaging was more accurate in determining the type of offending vessel than 3D-CISS or 3D-FLASH imaging. The retrospective study demonstrates that the combination of 3D-FLASH with 3D-CISS sequence well delineates the relationship between intracisternal segment of trigeminal nerve and adjacent vessels in terms of increased positive rates and accuracy. Advances in knowledge: The study firstly dealt with the combination of 3D-CISS and 3D-FLASH imaging in TN.


Relapse of Pneumocystis carinii pneumonia in the upper lobes during aerosol pentamidine prophylaxis.
Pneumocystis carinii pneumonia was diagnosed by bronchoalveolar lavage of the upper lobes in a patient with the acquired immunodeficiency syndrome (AIDS) receiving aerosol pentamidine prophylaxis. Serendipitous availability of a normal premorbid lung gallium scan indicated that pneumocystosis had developed during aerosol pentamidine prophylaxis; at the time of presentation a repeat gallium scan indicated disease limited to the upper lobes. The relation of this unusual form of isolated upper lobe Pneumocystis carinii pneumonia to aerosol pentamidine prophylaxis warrants further investigation.


Laryngopharyngeal reflux in patients with symptoms of gastroesophageal reflux disease.
Laryngopharyngeal reflux (LPR) has been extensively studied in patients with laryngeal signs and symptoms, gastroesophageal reflux being identified in approximately 50%. Few studies have investigated the incidence and significance of LPR in GERD patients. Two-hundred and seventy-six consecutive patients referred with symptoms of gastroesophageal reflux had dual probe 24 h pH, esophageal manometry, GERD and ENT questionnaires. LPR was defined as at least three pharyngeal reflux events less than pH 5.0 with corresponding esophageal reflux, but excluding meal periods. Fourty-two percent of patients were positive for LPR on 24 h pH monitoring and 91.3% corresponded with an abnormal esophageal acid score. Distal esophageal acid exposure was significantly greater (P < 0.001) in patients with LPR but symptoms of GERD and regurgitation scores showed no significant differences between patients with positive and negative LPR on 24 h pH. There was no significant difference between the incidence of LPR in patients with or without laryngeal symptoms. There is a high incidence of LPR in patients with GERD but its significance for laryngeal symptoms is tenuous. Fixed distance dual probe pH monitoring allows documentation of conventional esophageal reflux and LPR.


Natural Killer Cells Expanded and Preactivated Exhibit Enhanced Antitumor Activity against Different Tumor Cells in Vitro.
One of the emerging treatment strategies for cancer particularly for haematological malignancies is natural killer (NK) cell therapy. However, the availability of a best approach to maximize NK cell anticancer potential is still awaited. It is well established that cytokine-induced memory-like NK cells have the potential to differentiate after a short period of preactivation with interleukins-IL-12, IL-15, and IL-18 and exhibit increased responses to cytokine or activating receptor restimulation for weeks to months after preactivation. We demonstrated that NK cells differentiated from CD34+ cells isolated from cord blood show increased antitumor potential in vitro against different cancer cells. Using flow cytometry, we found that NK cells were able to induce apoptosis in cancer cells in vitro. We further analysed surviving gene expression by quantitative real time PCR and reported that NK cells cause down regulation of survivin gene expression in tumor cells. Therefore, NK cell therapy represents a promising immunotherapy for cancers like AML and other haematological malignancies. It concluded that NK cells can be differentiated from CD34+ cells isolated from cord blood ,are able to induce apoptosis and induce increased antitumor potential in vitro against different cancer cells besides cause downregulation of survivin gene expression in tumor cells. Therefore, NK cell therapy represents a promising immunotherapy for different cancer types and haematological malignancies. Furthers studies are necessary to confirm our findings.<br />.


Addition of four doses of rituximab to standard induction chemotherapy in adult patients with precursor B-cell acute lymphoblastic leukaemia (UKALL14): a phase 3, multicentre, randomised controlled trial.
Treatment for adults with acute lymphoblastic leukaemia requires improvement. UKALL14 was a UK National Cancer Research Institute Adult ALL group study that aimed to determine the benefit of adding the anti-CD20 monoclonal antibody, rituximab, to the therapy of adults with de novo B-precursor acute lymphoblastic leukaemia. This was an investigator-initiated, phase 3, randomised controlled trial done in all UK National Health Service Centres treating patients with acute lymphoblastic leukaemia (65 centres). Patients were aged 25-65 years with de-novo BCR-ABL1-negative acute lymphoblastic leukaemia. Patients with de-novo BCR-ABL1-positive acute lymphoblastic leukaemia were eligible if they were aged 19-65 years. Participants were randomly assigned (1:1) to standard-of-care induction therapy or standard-of-care induction therapy plus four doses of intravenous rituximab (375 mg/m2 on days 3, 10, 17, and 24). Randomisation used minimisation and was stratified by sex, age, and white blood cell count. No masking was used for patients, clinicians, or staff (including the trial statistician), although the central laboratory analysing minimal residual disease and CD20 was masked to treatment allocation. The primary endpoint was event-free survival in the intention-to-treat population. Safety was assessed in all participants who started trial treatment. This study is registered with ClincialTrials.gov, NCT01085617. Between April 19, 2012, and July 10, 2017, 586 patients were randomly assigned to standard of care (n=292) or standard of care plus rituximab (n=294). Nine patients were excluded from the final analysis due to misdiagnosis (standard of care n=4, standard of care plus rituximab n=5). In the standard-of-care group, median age was 45 years (IQR 22-65), 159 (55%) of 292 participants were male, 128 (44%) were female, one (<1%) was intersex, and 143 (59%) of 244 participants had high-risk cytogenetics. In the standard-of-care plus rituximab group, median age was 46 years (IQR 23-65), 159 (55%) of 294 participants were male, 130 (45%) were female, and 140 (60%) of 235 participants had high-risk cytogenetics. After a median follow-up of 53·7 months (IQR 40·3-70·4), 3-year event-free survival was 43·7% (95% CI 37·8-49·5) for standard of care versus 51·4% (45·4-57·1) for standard of care plus rituximab (hazard ratio [HR] 0·85 [95% CI 0·69-1·06]; p=0·14). The most common adverse events were infections and cytopenias, with no difference between the groups in the rates of adverse events. There were 11 (4%) fatal (grade 5) events in induction phases 1 and 2 in the standard-of-care group and 13 (5%) events in the standard-of-care plus rituximab group). 3-year non-relapse mortality was 23·7% (95% CI 19·0-29·4) in the standard-of-care group versus 20·6% (16·2-25·9) in the standard-of-care plus rituximab group (HR 0·88 [95% CI 0·62-1·26]; p=0·49). Standard of care plus four doses of rituximab did not significantly improve event-free survival over standard of care. Rituximab is beneficial in acute lymphoblastic leukaemia but four doses during induction is likely to be insufficient. Cancer Research UK and Blood Cancer UK.


Risk of non-tumoural portal vein thrombosis in patients with HCV-induced cirrhosis after sustained virological response.
Sustained virological response (SVR) to direct-acting antivirals ameliorates portal hypertension, improves hepatic function and may reverse the procoagulant state observed in patients with cirrhosis. However, an unexpected incidence of portal vein thrombosis (PVT) immediately after antiviral therapy has recently been reported. Therefore, we analysed the long-term impact of SVR on the development of non-tumoural PVT. Our study comprised of two well-characterized prospective cohorts (hepatitis C virus '(HCV)-Cured': n = 354/'HCV-Active': n = 179) of patients with HCV cirrhosis who underwent standardized ultrasound surveillance. In the main analysis, the event of interest was de novo non-tumoural PVT and events known to modify its natural history (orthotopic liver transplantation, transjugular intrahepatic portosystemic shunt, death, tumoural PVT and anticoagulation) were considered as competing risk. Adjusted models were built using propensity scores for baseline covariates. Moreover, predictive factors were investigated by conventional multivariate analysis. Ten (2.8%) patients in the 'HCV-Cured' cohort developed a non-tumoural PVT during a median follow-up of 37.1 months, while 8 (4.5%) patients in the 'HCV-Active' cohort were diagnosed with non-tumoural PVT during a median follow-up of 42.2 months. High Child-Pugh score was the only independent risk factor for non-tumoural PVT development and stage A patients were at low risk. Importantly, HCV cure did not decrease the risk of non-tumoural PVT in inverse probability of treatment-weighted (IPTW) analysis (subdistribution hazard ratio: 1.31 (95% confidence interval [95% CI]: 0.43-3.97); P = .635). In contrast, SVR was associated with a substantial reduction in mortality (IPTW-adjusted sHR: 0.453 [95% CI: 0.287-0.715]; P < .001). The risk of non-tumoural PVT persists after HCV cure in patients with cirrhosis, despite improving survival. Even after aetiological cure, severity of liver disease remains the main determinant of non-tumoural PVT development.


Stepwise intramolecular photoisomerization of NHC-chelate dimesitylboron compounds with C-C bond formation and C-H bond insertion.
C,C-chelate dimesitylboron (BMes(2)) compounds containing an N-heterocyclic carbene (NHC) donor have been obtained. Single-crystal X-ray diffraction analyses established that the boron atom in these compounds is bound by four carbon atoms in a distorted tetrahedral geometry. Compared to previously reported N,C-chelate dimesitylboron compounds, the new C,C-chelate boron compounds have a much larger HOMO-LUMO energy gap (>3.60 eV). They do, however, respond to UV irradiation (300 nm) in the same manner as N,C-chelate BMes(2) compounds do, undergoing photoisomerization and converting to an intensely colored (yellow or orange) isomer A quantitatively, with a high quantum efficiency (0.60-0.75). NMR and single-crystal X-ray diffraction analyses established that the structure of A is similar to the dark isomers obtained from N,C-chelate BMes(2) compounds. However, unlike the N,C-chelate dark isomers that have the tendency to thermally reverse back to the light colored isomers, the isomers A of the C,C-chelate BMes(2) are thermally stable and no reverse isomerization was observed even when heated to 80 °C (or 110 °C) for hours. The most unusual finding is that isomers A undergo further photoisomerization when irradiated at 350 nm, forming a new colorless species B nearly quantitatively. NMR and single-crystal X-ray diffraction analyses established the structure of isomer B, which may be considered as an intramolecular C-H insertion product via a borylene intermediate. Mechanistic aspects of this unusual two-step photoisomerization process have been examined by DFT computational studies.


Luteolytic influence of intrauterine dead embryos in the early pregnant rat.
The present investigation was an endeavour to study if annihilation of embryos in the uterus of the rat before the establishment of placental luteotropic functions has any influence on corpus luteal function, and, if there is any, whether it is local or systemic. The responsibility of pregnancy maintenance was imposed on a single ovary by performing unilateral ovariectomy after implantation (on Day 5 postcoitum). The implantation sites in one uterine horn, either ipsilateral or contralateral to the remaining ovary, were selectively destroyed by injecting 0.1 ml of sterile normal saline to that particular horn only, and the peripheral progesterone level and viability of the embryos in the untreated horn, which depended on the functions of the remaining ovary, were examined. Selective killing of embryos in the uterine horn of the ovariectomized side did not exert any influence on the fetal viability in the untreated horn ( nonovariectomized side) and the peripheral progesterone level also remained statistically unaffected. On the contrary, induction of fetal resorptions in the uterine horn of the intact side produced a significant fall in the peripheral level of progesterone and induced resorption of embryos of the ovariectomized side also. The latter could significantly be prevented by simultaneous administration of exogenous progesterone, indicating luteolysis as the major, if not sole, factor responsible for fetal resorption in the untreated horn. The luteolytic effect was attributed neither to saline itself, nor to the distension of the uterine horn caused by saline injection. Luteolytic factors from the dead embryo-bearing horn which act locally on the adjacent ovary only, are discussed.


Erythematous candidiasis leading to systemic manifestations of human immunodeficiency virus co-infection with secondary syphilis: A diagnostic and therapeutic dilemma.
The intensification of human immunodeficiency virus (HIV) and rising frequency of immunocompromised individuals have resulted in a resurgence of opportunistic infections. The most common opportunistic oral fungal infection in HIV-positive individuals is oral candidiasis. The classical presentation is as white scrapable form called as thrush, which is easily diagnosed and treated. The clinician is presented with a diagnostic and management dilemma when these lesions appear in new facades such as erythematous candidiasis, the latter's prevalence with HIV and AIDS being well established. In this case report, we present a case of Erythematous Candidiasis, which was associated with type 1 HIV co-infected with syphilis. We highlight the diagnostic importance of a naive looking manifestation of the tongue which was followed by a series of challenging presentations of secondary syphilis. Since the patient had a negative Veneral Disease Research Laboratory and left us with a management dilemma, the article also features the importance of prozone phenomenon (seen in 2% cases of secondary syphilis), and it's higher association with HIV co-infected individuals. With confusing clinical oral manifestations associated with these diseases, the dentist might be the first person to encounter such lesions, who should be able to recognize erythematous candidiasis and correlate them with the underlying pathology.


A nonlinear electrical-thermal model of the skin.
This work presents a model for the skin which accounts for both the nonlinearities and the asymmetries in its voltage-current characteristic. This model consists of an electrical submodel and a heat transfer submodel. The electrical submodel uses nonlinear devices in which some parameters depend on skin temperature. The heat transfer submodel models the heat exchange between the skin, the surrounding tissues, and the ambient medium and calculates the temperature of the skin to update the necessary parameters of the electrical submodel. The model is based on experiments designed to determine: 1) the dry skin voltage-current characteristic; 2) the changes in the skin breakdown voltage with location; 3) the moist skin voltage-current characteristic; 4) the changes in the voltage-current characteristic of the skin with duration after the onset of stimulation; and 5) the effect of skin temperature on its voltage-current characteristic. During these experiments we used 84-mm2 square Ag-AgCl electrodes to apply sinusoidal voltage of 0.2 and 20 Hz. The simulations were performed using the Advanced Continuous Simulation Language (ACSL), capable of solving differential and integral equations with variable coefficients. The model predicted the skin behavior satisfactorily for a large range of amplitudes and frequencies. We found that the breakdown occurred when the energy delivered to the skin exceeded a threshold. Above this threshold the voltage-current characteristic of the skin became nonlinear and asymmetric and, in a real situation, the subject would experience an uncomfortable sensation which could rapidly develop into pain.


Acute effects of kinesio taping on muscular strength and endurance parameters of the finger flexors in sport climbing: A randomised, controlled crossover trial.
Kinesio taping (KT) is a commonly used intervention in sports and, recently, KT has become popular among athletes competing in sport climbing and bouldering events. However, evidence on the effect of KT on grip strength and endurance is still controversial. Therefore, the purpose of this study was to evaluate immediate effects of KT on muscular strength and endurance of the finger flexor muscles in sport climbers. Twenty recreationally-trained active sport climbers (10 men, 10 women) aged 28.5 ± 10.6 years performed one familiarisation trial and subsequently, in a randomised crossover design, two test trials either with (TAPE) or without (CONTROL) KT over the finger flexor muscles. Test trials consisted of three performance measurements (hand grip strength and endurance, finger hang, and lap climbing) at intervals of 48 h in a randomised order. We observed no significant differences in the parameters of hand grip peak force, fatigue index, finger hang time, lap climbing distance and time, or maximum blood lactate values after lap climbing between the TAPE and CONTROL trials (p > 0.05). The participants' climbing ability was significantly correlated with the intra-individual performance changes between the TAPE and CONTROL conditions for the fatigue index (r = -0.598, p = 0.005), but not in any of the other performance-related parameters. Therefore, KT over the finger flexor muscles neither enhanced hand grip strength and endurance nor the sport climbing performance parameters of finger hang, lap climbing distance and time, and maximum blood lactate values after lap climbing.


Membrane lipid regulation in Acholeplasma laidlawii grown with saturated fatty acids. Biosynthesis of a triacylglucolipid forming reversed micelles.
The membrane lipid composition in several strains of Acholeplasma laidlawii is regulated upon a change in the growth conditions. Monoglucosyldiacylglycerol (MGlcDAG) and diglucosyldiacylglycerol (DGlcDAG) are the most abundant lipids in the A. laidlawii membrane. A third glucolipid, 3-O-acyl-monoglucosyldiacylglycerol (MAMGlcDAG) is synthesized by strain A-EF22 when the membrane lipids contain large amounts of saturated acyl chains. The lipid regulation can be understood from a simple theoretical model, in which the cells strive to maintain a balance between the lipids constituting a bilayer and those forming reversed non-lamellar liquid crystalline phases. Thus, the physical chemistry of membrane lipids, in particular their ability to form different aggregate structures, constitutes the basis for the lipid regulation, and therefore an understanding of the phase equilibria of membrane lipids is crucial. MGlcDAG and MAMGlcDAG isolated from A. laidlawii strain A-EF22 membranes were studied mainly by 2H NMR, 1H NMR, and 1H NMR diffusion measurements. MAMGlcDAG, containing 96 mol % saturated acyl chains formed a gel/crystalline phase up to about 80 degrees C, where a transition occurred to a reversed micellar (L2) phase. This is an unexpected finding for a membrane lipid. However, this lipid homogeneously mixes with the other membrane lipids at physiological temperatures. Previous and new data on MGlcDAG show that the lamellar phase is stabilized when the length and the degree of unsaturation of the acyl chains are decreased. The physicochemical properties of MAMGlcDAG and MGlcDAG were compared and found to be of great significance for the physiological regulation of the lipids in the membrane. MAMGlcDAG is synthesized under conditions when the phase equilibria of MGlcDAG are shifted from a non-lamellar toward a lamellar phase. Apart from MAMGlcDAG, MGlcDAG is the major lipid in A. laidlawii strain A-EF22 which is able to form reversed aggregate structures. MAMGlcDAG probably assists MGlcDAG in maintaining an optimal molecular packing, or negative curvature, of the lipids in the membrane.


Low number of peripheral blood B lymphocytes in patients with pulmonary tuberculosis.
The cellular immune response plays a critical role in the containment of persistent Mycobacterium tuberculosis infection; however, the immunological mechanisms that lead to its control are not completely identified. The goal of this study was to evaluate B (CD19+) and T (CD3+) peripheral blood lymphocyte profiles and T-cell subsets (CD4+ and CD8+) in patients with pulmonary tuberculosis (TB). Percentages (p = 0.02) and absolute numbers (p = 0.005) of B cells were significantly lower in patients with pulmonary TB than in healthy donors. In contrast, percentages (p = 0.12) and absolute numbers (p = 0.14) of T cells were similar in TB patients and healthy donors. No significant differences in percentages of CD4+ (p = 0.19) or CD8+ (p = 0.85) T cells between patients and healthy donors were observed. In summary, patients with pulmonary tuberculosis had a lower number of peripheral blood B lymphocytes than healthy controls.


The ATP-dependent ClpXP and Lon proteases regulate expression of the Yersinia pestis type III secretion system via regulated proteolysis of YmoA, a small histone-like protein.
The Yersinia pestis plasmid pCD1-encoded type III secretion system (T3SS) is essential for the pathogenicity of Y. pestis in mammalian hosts. T3SS-associated genes are maximally expressed at 37 degrees C in the absence of extracellular calcium. Expression of T3SS genes requires LcrF, an AraC-like transcriptional activator, and is repressed by YmoA, a small histone-like protein. The mechanism by which temperature regulates T3SS gene expression has not been determined; however, changes in DNA topology have been implicated in this process. We report here that a Y. pestis strain deficient in production of the ClpXP and Lon proteases does not express a functional T3SS partly because of high cytosolic levels of YmoA. YmoA is rapidly degraded at 37 degrees C in wild-type Y. pestis, but remains stable in a clpXPlon deletion mutant. The stability of YmoA in wild-type Y. pestis increased as the growth temperature of the culture decreased; in contrast, YmoA was stable at all temperatures examined in the clpXPlon deletion mutant. These results indicate that the ClpXP and Lon proteases contribute to the environmental regulation of the Y. pestis T3SS system through regulated proteolysis of YmoA.


A nuclear antigen associated with cell proliferation and blast transformation.
A nuclear antigen associated with cell proliferation (proliferating cell nuclear antigen-PCNA) and blast transformation is recognized by autoantibodies in the sera of some patients with systemic lupus erythematosus. This autoantibody is a precipitating antibody and also reacts in immunofluorescence, staining the nucleoplasm of proliferating and blast-transformed cells. The autoantibody was used as a reagent to determine the distribution of PCNA in a synchronized continuous B lymphoid cell line (WiL-2) and in mitogen-induced blast-transformed lymphocytes. In WiL-2 cells, PCNA was detected as speckled nucleoplasmic staining in G1, S, and G2 phases of the cell cycle. In addition, during late G1 and early S phases, PCNA was also detected in the nucleolus. During mitogen-induced blast transformation of lymphocytes, PCNA was noticed in the nucleolus before the initiation of DNA synthesis and later became nucleoplasmic with disappearance of nucleolar staining. These studies demonstrate that the relationship of PCNA to proliferation and blast transformation may be associated with events related to DNA synthesis in these cells.


Detection of P-glycoprotein activity in endotoxemic rats by 99mTc-sestamibi imaging.
(99m)Tc-sestamibi is a widely used radiopharmaceutical agent for myocardial and oncologic imaging. Because of its unique role as a P-glycoprotein (Pgp)-specific substrate, this compound can be used to examine Pgp functional activity in vitro and in vivo under pathologic conditions. Our objective was to use (99m)Tc-sestamibi as a tool to investigate whether systemic inflammation induced by Escherichia coli lipopolysaccharide (LPS) would affect in vivo Pgp function in the brain, heart, liver, and kidneys of rats. Moreover, we also wanted to examine LPS-mediated effects in the placenta of pregnant rats because of the limited amount of in vivo data on this tissue. Rats were injected intraperitoneally with LPS or an equal volume of saline as controls. After certain time periods (6 or 24 h), animals were administered 20 MBq of (99m)Tc-sestamibi intravenously, and then images were taken at 0.5, 1, 2, and 3 h. Tissues of rats were excised for (99m)Tc-sestamibi biodistribution analysis by gamma-counting and messenger RNA (mRNA) analysis by reverse transcription-polymerase chain reaction. Western blot analysis with antibody C-219 was used to detect Pgp levels. LPS treatment for 6 h caused a significant downregulation of mdr1a mRNA levels in the brain, heart, and liver, whereas 24 h of LPS treatment significantly reduced mdr1a mRNA levels only in the liver. A significant downregulation of mdr1a mRNA was seen in the brain, heart, and liver within 6 h after LPS administration. Imaging and biodistribution studies demonstrated a higher accumulation of (99m)Tc-sestamibi in the brain, heart, and liver of LPS-treated rats. In the brain, LPS-imposed downregulation of mdr1a mRNA levels was transient, with significant suppression at 4, 6, and 12 h, and the levels recovered to nearly normal by 24 h. This time-dependent downregulation of mRNA correlated with protein levels determined by Western blot analysis. Biodistribution studies of pregnant rats demonstrated a 3.5-fold-higher accumulation of (99m)Tc-sestamibi in the fetal tissues of LPS-treated pregnant rats than in saline-treated control rats. Furthermore, placental mdr1a and mdr1b mRNA levels were also significantly downregulated by LPS treatment. Our results indicate that LPS-induced systemic inflammation caused an increased retention of (99m)Tc-sestamibi in the brain, heart, liver, and fetal tissues. These results correlated with a reduction in mdr1a mRNA levels in each organ.


Proponent or collaborative: Physician perspectives and approaches to disease modifying therapies in sickle cell disease.
Sickle cell disease (SCD) is an inherited blood disorder that primarily affects African-American and other ethnic minority populations. There are three available disease-modifying therapies for sickle cell disease: hydroxyurea (HU), bone marrow transplantation (BMT), and chronic blood transfusion (CBT). Since these treatments vary in their therapeutic intent, efficacy in preventing progression of the disease, short and long-term adverse effects, costs and patient burden, the decision-making process regarding these therapies is complicated for both the patient and healthcare provider. While previous research has focused on the patient perspective of treatment-related decision making, there is a paucity of research investigating the physician perspective of treatment-related decision making. We conducted a qualitative study with physicians who were experts in the field of SCD. Interviews focused on physician perceptions of patient decisional needs as well as physicians' approach to decision making regarding disease-modifying therapies in SCD. Thirty-six physician interviews were analyzed, with a focus on their perspectives regarding available treatment options and on how they approach decision making with patients. We identified two narrative approaches. The Collaborative approach (CA) was characterized by emphasizing the need to discuss all possible treatment options to ensure that the patient and/or family was equipped to make an informed decision. The Proponent approach (PA) was characterized by strongly advocating a pre-determined treatment plan and providing patients/families with information, with the objective of convincing them to accept the treatment. An interplay of patient-related and disease-related factors, decision type and physician-related factors, as well as institutional frameworks, influenced physician perspectives on treatment options and decision making regarding these therapies. These findings point to the potential value of developing systems to foster patient engagement as a way of facilitating shared decision making.


Medialisation laryngoplasty: a new surgical frontier in the management of vocal cord paralysis.
Patients who are victims of vocal cord paralysis usually suffer from husky or hoarse voice which is very much jeopardising their future particularly in professional voice users like singers, priests, teachers, etc. Phonosurgical techniques like medialisation laryngoplasty has opened a new horizon in the management of dysphonia with unilateral vocal cord paralysis. Medialisation laryngoplasty is sometimes described under the heading of laryngoplastic phonosurgery or thyroplasty. A report of 10 cases of unilateral vocal cord paralysis with dysphonia subjected to medialisation laryngoplasty from March 1996 to March 1998 with 90% success is presented. Only one patient developed inspiratory stridor as a complication.


Abdominal crisis in a young man with systemic lupus erythematosus.
Medium-sized artery aneurysms are rare in patients with systemic lupus erythematosus (SLE). We report on a 21-year-old Chinese man with SLE and secondary antiphospholipid syndrome (APS) who presented with acute abdominal pain due to a ruptured right hepatic artery aneurysm. He was also found to have aneurysms of the left hepatic artery and splenic artery on autopsy. There have been only eight cases of hepatic artery aneurysm and one case of splenic artery aneurysm associated with SLE in the English literature. Abdominal aneurysm must be suspected in SLE patients presenting with acute abdominal pain, haemoperitoneum or occult bleeding.


Physiological concentrations of nitric oxide do not elicit an acute negative inotropic effect in unstimulated cardiac muscle.
We examined the effect of several nitric oxide (NO) donors, authentic NO gas, and L-arginine in isolated cat and rat papillary muscles. We did not observe significant inotropic effects in response to any NO donor (ie, SPM-5185, C87-3754, and S-nitroso-N-acetylpenicillamine [SNAP]) from 1 nmol/L to 100 mumol/L. Similarly, authentic NO, at concentrations far in excess of those that maximally dilate the coronary vasculature (ie, 500 nmol/L), also failed to exert a detectable inotropic effect in these preparations. However, in the presence of 5 mumol/L norepinephrine, 500 nmol/L NO exerted a 12 +/- 3% decrease in isolated rat papillary muscle contractility (P < .05). Addition of L-arginine up to 25 mmol/L exerted no inotropic effects in isolated rat papillary muscles. However, at 50 mmol/L, L-arginine decreased contractile force by 21 +/- 4% (P < .01). On further examination, the negative inotropic effect of 50 mmol/L L-arginine appeared to be nonspecific, since the inactive stereoisomer, D-arginine, at 50 mmol/L exerted the same effect. Further studies in isolated adult rat cardiac myocytes elicited similar results, in that 50 mmol/L of L- and D-arginine equally decreased contraction amplitude and the underlying cytosolic calcium transient. Moreover, 500 nmol/L of the NO donor SPM-5185 only modestly decreased contraction amplitude or intracellular calcium in isolated rat cardiac myocytes. These results indicate that administration of physiological concentrations of exogenous NO does not acutely depress the inotropic state of the rat or cat heart to a physiologically significant extent.(ABSTRACT TRUNCATED AT 250 WORDS)


The confusing array of tooth-colored crowns.
The venerable PFM crown or fixed prosthesis still dominates the tooth-colored restoration market. However, use of PFMs is declining slightly, as the many new all-ceramic and resin-based composite crowns and fixed-prosthesis products flood the market. Several situations may indicate the use of materials other than PFM. They include patients requiring a high level of esthetic acceptability, patients with proven or perceived allergies to the metals used in dentistry and bruxing or clenching patients with metal allergies or desire to eliminate metal from their mouths. PFM restorations have been proven during 40 years of successful use. They provide acceptable esthetics for most situations, minimal fracture during service, proven ability to serve in multiple-unit situations and excellent fit, and the profession has detailed knowledge of these restorations' advantages, disadvantages and physical characteristics. PFM restorations have only a few well-known negative characteristics. Be cautious as you elect to move from the reliability and positive history of PFM to relatively unknown types of restorations.


Neuropeptide Y Y4 receptor homodimers dissociate upon agonist stimulation.
The pancreatic polypeptide-fold family of peptides consists of three 36-amino acid peptides, namely neuropeptide Y (NPY), peptide YY, and pancreatic polypeptide (PP). These peptides regulate important functions, including food intake, circadian rhythms, mood, blood pressure, intestinal secretion, and gut motility, through four receptors: Y1, Y2, Y4, and Y5. Additional receptor subtypes have been proposed based on pharmacology observed in native tissues. Recent studies with other G-protein-coupled receptors have shown that homo- and heterodimerization may be important in determining receptor function and pharmacology. In the present study, the recently cloned rhesus (rh) Y4 receptor was evaluated using radioligand binding, and the pharmacological profile was found to be very similar to the human Y4 receptor. To study homo- and heterodimerization involving the Y4 receptor using bioluminescence resonance energy transfer 2 (BRET(2)), the carboxy termini of the rhesus Y1, Y2, Y4, and Y5 receptors were fused to Renilla luciferase, and rhY4 was also fused to green fluorescent protein. Dimerization was also studied using Western blot analysis. Using both BRET(2) and Western analysis, we found that the rhY4 receptor is present at the cell surface as a homodimer. Furthermore, agonist stimulation using the Y4-selective agonists PP and 1229U91 can dissociate these dimers in a concentration-dependent manner. In contrast, rhY4 did not heterodimerize with other members of the NPY receptor family or with human opioid delta and mu receptors. Therefore, homodimerization is an important component in the regulation of the Y4 receptor.


Development and multicenter validation of a multiparametric imaging model to predict treatment response in rectal cancer.
To develop and validate a multiparametric model to predict neoadjuvant treatment response in rectal cancer at baseline using a heterogeneous multicenter MRI dataset. Baseline staging MRIs (T2W (T2-weighted)-MRI, diffusion-weighted imaging (DWI) / apparent diffusion coefficient (ADC)) of 509 patients (9 centres) treated with neoadjuvant chemoradiotherapy (CRT) were collected. Response was defined as (1) complete versus incomplete response, or (2) good (Mandard tumor regression grade (TRG) 1-2) versus poor response (TRG3-5). Prediction models were developed using combinations of the following variable groups: (1) Non-imaging: age/sex/tumor-location/tumor-morphology/CRT-surgery interval (2) Basic staging: cT-stage/cN-stage/mesorectal fascia involvement, derived from (2a) original staging reports, or (2b) expert re-evaluation (3) Advanced staging: variables from 2b combined with cTN-substaging/invasion depth/extramural vascular invasion/tumor length (4) Quantitative imaging: tumour volume + first-order histogram features (from T2W-MRI and DWI/ADC) Models were developed with data from 6 centers (n = 412) using logistic regression with the Least Absolute Shrinkage and Selector Operator (LASSO) feature selection, internally validated using repeated (n = 100) random hold-out validation, and externally validated using data from 3 centers (n = 97). After external validation, the best model (including non-imaging and advanced staging variables) achieved an area under the curve of 0.60 (95%CI=0.48-0.72) to predict complete response and 0.65 (95%CI=0.53-0.76) to predict a good response. Quantitative variables did not improve model performance. Basic staging variables consistently achieved lower performance compared to advanced staging variables. Overall model performance was moderate. Best results were obtained using advanced staging variables, highlighting the importance of good-quality staging according to current guidelines. Quantitative imaging features had no added value (in this heterogeneous dataset). Predicting tumour response at baseline could aid in tailoring neoadjuvant therapies for rectal cancer. This study shows that image-based prediction models are promising, though are negatively affected by variations in staging quality and MRI acquisition, urging the need for harmonization. This multicenter study combining clinical information and features derived from MRI rendered disappointing performance to predict response to neoadjuvant treatment in rectal cancer. Best results were obtained with the combination of clinical baseline information and state-of-the-art image-based staging variables, highlighting the importance of good quality staging according to current guidelines and staging templates. No added value was found for quantitative imaging features in this multicenter retrospective study. This is likely related to acquisition variations, which is a major problem for feature reproducibility and thus model generalizability.


Serum vascular endothelial growth factor as a possible indicator of arterial reactivity in postmenopausal women.
To determine whether serum vascular endothelial growth factor (VEGF) concentration is correlated with arterial reactivity in postmenopausal women. An observational study was conducted on 24 postmenopausal women who did not receive any hormone therapy in the 6 months preceding the study and had no pre-existing cardiovascular diseases or cardiovascular risk factors. Serum samples were obtained from all participants and analyzed for VEGF concentrations. Arterial reactivity was assessed by the measurement of endothelium-dependent dilatation and endothelium-independent dilatation of the brachial artery, using color duplex Doppler ultrasound. The study population was aged 50.8 years on average, with about 2 years of menopause. Serum VEGF concentration in postmenopausal women was significantly correlated with both endothelium-dependent dilatation (r = -0.66, p < 0.01) and endothelium-independent dilatation (r = -0.65, p < 0.01) of the brachial artery. Serum VEGF level may be a potential indicator of arterial reactivity in postmenopausal women.


Physcion prevents induction of optic nerve injury in rats via inhibition of the JAK2/STAT3 pathway.
Optic nerve injury is a type of neurodegenerative disease. Physcion is an anthraquinone that exerts a protective role against various diseases. However, its function in regulating optic nerve injury remains largely unknown. An in vitro model of optic nerve injury was established in HAPI cells treated with IFN-β. Functional assays were used to detect HAPI cell viability and apoptosis. The levels of inflammation and the expression levels of oxidative stress-related genes were measured in HAPI cells. In addition, western blot analysis was used to detect the expression levels of Janus kinase 2 (JAK2)/STAT3-linked genes in HAPI cells. Treatment of the cells with physcion prevented cells against IFN-β-induced neuronal injury. Physcion restrained IFN-β-induced inflammatory response and oxidative stress in HAPI cells. In addition, it improved IFN-β-induced injury in HAPI cells by suppressing the JAK2/STAT3 pathway. In conclusion, the present study revealed that physcion improved optic nerve injury in vitro by inhibiting the JAK2/STAT3 pathway. Physcion may be a promising therapeutic target for the treatment of this disease.


Failure of puberty and linear growth in beta-thalassemia major.
Thalassemia major is a severe progressive hemolytic anemia and a serious medical problem worldwide. Endocrine dysfunctions are well described in patients with thalassemia major. Data for endocrine complications from developing countries are scant. Endocrine complications in developing countries may be frequent due to suboptimal iron chelation. The aim of this study was to evaluate the prevalence of delayed puberty and growth failure in patients with beta-thalassemia major. We evaluated the growth and sexual development of 146 patients with thalassemia major aged 10-22 years. The following data were recorded in questionnaire, age, sex, height, weight, serum ferritin levels and pubertal staging. Failure of puberty was present in 75.6% of boys and 68.4% of girls aged 12-22 years. Gonadotropin insufficiency was found in most of the patients with lack of puberty. There was a significant difference between the height of patients with pubertal development (153 ± 9.1 cm) and those with delayed puberty (140 ± 9.1), (p< 0.001). Short stature was present in 65.7% of patients. Sixty-nine percent of boys and 62.9% of girls were found to be less than 2 SD below the mean for normal height; after the age of 12, the percentage was 78.9% in girls and 83.8% in boys after the age of 14. We conclude that failure of puberty and impaired growth are very common in our thalassemic patients which necessitates newer protocols of treatment, correct blood transfusion and chelation therapy.


Respiratory sinus arrhythmia as a surrogate measure of respiratory frequency: validity and robustness to activity in rats.
Recording of breathing frequency is a basic requirement for respiratory physiology. Usual techniques are invasive and constraining. Respiratory sinus arrhythmia (RSA) has recently been demonstrated to be a simple way to obtain respiration frequency at rest. In this study, we investigated whether this correlation is also observed during activity. We first compared RSA to the respiration frequency obtained in anesthetized rats using a pneumotachograph connected to the trachea (TRF). Data analyses using Passing and Bablok regression confirmed the absence of bias and proportional differences. Accordingly, the Bland-Altman plot did not show any significant differences in data sets. In a second experiment, we compared RSA to the respiration frequency obtained in freely moving rats using a subpleurally inserted telemetric catheter (PRF). Comparisons between RSA and PRF revealed no significant difference in determination of respiratory rate with the two methods, although the bias and confidence interval were greater when activity increased. This was, however, not the case during short episodes of sniffing-like tachypnea, during which no matching RSA peaks were observed. In conclusion, RSA frequency reflected regular respiration frequency independently of the level of activity and appears to be a good surrogate to usual techniques.


Insulin resistance--a risk factor for coronary heart disease?
Fasting insulin secretion was assessed by measuring fasting serum C-peptide levels in 529 women and 399 men aged 18-90 years, to study the relationship between insulin secretion, insulin resistance and risk factors for coronary heart disease. Subjects with low serum high density lipoprotein (HDL) cholesterol levels showed higher mean serum insulin and C-peptide levels than subjects with normal HDL cholesterol levels. In male subjects these differences were significant for both serum insulin and serum C-peptide results (P less than 0.005). In female subjects serum insulin results differed significantly (P less than 0.0005) but for the difference in mean serum C-peptide levels P was equal to 0.012. Fasting serum C-peptide correlated negatively with serum HDL cholesterol. However, serum C-peptide also correlated with serum triglyceride and serum triglyceride correlated negatively with serum HDL cholesterol. Each correlation was statistically significant (P less than 0.001). Multiple regression analysis suggested that the apparent association of C-peptide with HDL cholesterol was a consequence of the interrelated association between C-peptide, triglyceride and HDL cholesterol. The analysis was consistent with the hypothesis that obesity and increased insulin resistance were associated with increased insulin secretion and in turn with high serum triglyceride levels and consequentially low levels of serum HDL cholesterol. The data were compatible with the suggestion that insulin resistance rather than fasting insulin concentration per se could be a risk factor for coronary heart disease.


[Effect of interleukin-6 promoter DNA methylation on the pathogenesis of systemic lupus erythematosus].
Objective: To analyze peripheral blood interleukin-6 (IL-6) promoter DNA methylation status and its clinical significance in patients with systemic lupus erythematosus (SLE). Methods: Blood samples of 41 adult patients with SLE and 20 healthy controls were collected.The methylation status of IL-6 promoter was determined by methylation specific polymerase chain reaction (MSP). The IL-6 expression was detected by real-time PCR.Correlations between IL-6 promoter methylation status and clinical features or laboratory findings in patients with SLE were analyzed. Results: The levels of IL-6 mRNA were significantly higher in peripheral blood of SLE.DNA methylation levels of IL-6 promoter were reduced in SLE patients as compared with healthy controls.The methylation status and expression of IL-6 in peripheral blood reflected the levels in peripheral blood mononuclear cells (PBMCs). Significantly positive correlation was found between IL-6 hypomethylation and renal disorder, as well as hypocomplementemia in patients with SLE. Conclusion: Hypomethylation of interleukin-6 promoter in peripheral blood might be involved in the etiology of SLE.


Contrast agents for hepatic MRI.
Liver specific contrast media (LSCM) can be subdivided according to different modalities of hepatic distribution: exclusive distribution to the hepatocellular compartment can be obtained using CM which accumulate within the hepatocytes after slow infusion; other CM demonstrate combined perfusion and hepatocyte-selective properties, with an initial distribution to the vascular-interstitial compartment (in an analogous manner to that of the conventional extracellular CM), thereafter, a fraction of the injected dose is taken up into the hepatocytes causing an increase in the signal intensity of the hepatic tissue. The use of the superparamagnetic effect of iron oxide particles is based on distribution in the reticuloendothelial system (RES), usually well represented in the normal parenchyma as well as in benign hepatocellular lesions, and absent in most malignant lesions. It is necessary to have an in-depth knowledge of either the biological and histological characteristics of focal liver lesions (FLL) or the enhancement mechanism of LSCM to gain significant accuracy in the differential diagnosis of FLL. Dynamic contrast-enhanced MRI is an important tool in the identification and characterization of FLL. With LSCM it is possible to differentiate benign from malignant lesions and hepatocellular lesions from non hepatocellular lesions with high accuracy. To understand the contrast behaviour after injection of LSCM it is necessary to correlate the contrast enhancement with both the biological and histological findings of FLL.


Detection of circulating tumor cells in breast cancer may improve through enrichment with anti-CD146.
Most assays to detect circulating tumor cells (CTCs) rely on EpCAM expression on tumor cells. Recently, our group reported that in contrast to other molecular breast cancer subtypes, "normal-like" cell lines lack EpCAM expression and are thus missed when CTCs are captured with EpCAM-based technology [J Natl Cancer Inst 101(1):61-66, 2009]. Here, the use of CD146 is introduced to detect EpCAM-negative CTCs, thereby improving CTC detection. CD146 and EpCAM expression were assessed in our panel of 41 breast cancer cell lines. Cells from 14 cell lines, 9 of which normal-like, were spiked into healthy donor blood. Using CellSearch technology, 7.5 ml whole blood was enriched for CTCs by adding ferrofluids loaded with antibodies against EpCAM and/or CD146 followed by staining for Cytokeratin and DAPI. Hematopoietic cells and circulating endothelial cells (CECs) were counterstained with CD45 and CD34, respectively. A similar approach was applied for blood samples of 20 advanced breast cancer patients. Eight of 9 normal-like breast cancer cell lines lacked EpCAM expression but did express CD146. Five of these 8 could be adequately recovered by anti-CD146 ferrofluids. Of 20 advanced breast cancer patients whose CTCs were enumerated with anti-EpCAM and anti-CD146 ferrofluids, 9 had CD146+ CTCs. Cells from breast cancer cell lines that lack EpCAM expression frequently express CD146 and can be recovered by anti-CD146 ferrofluids. CD146+ CTCs are present in the peripheral blood of breast cancer patients with advanced disease. Combined use of anti-CD146 and anti-EpCAM is likely to improve CTC detection in breast cancer patients.


Liver-infiltrating lymphocytes in chronic human hepatitis C virus infection display an exhausted phenotype with high levels of PD-1 and low levels of CD127 expression.
The majority of people infected with hepatitis C virus (HCV) fail to generate or maintain a T-cell response effective for viral clearance. Evidence from murine chronic viral infections shows that expression of the coinhibitory molecule PD-1 predicts CD8+ antiviral T-cell exhaustion and may contribute to inadequate pathogen control. To investigate whether human CD8+ T cells express PD-1 and demonstrate a dysfunctional phenotype during chronic HCV infection, peripheral and intrahepatic HCV-specific CD8+ T cells were examined. We found that in chronic HCV infection, peripheral HCV-specific T cells express high levels of PD-1 and that blockade of the PD-1/PD-L1 interaction led to an enhanced proliferative capacity. Importantly, intrahepatic HCV-specific T cells, in contrast to those in the periphery, express not only high levels of PD-1 but also decreased interleukin-7 receptor alpha (CD127), an exhausted phenotype that was HCV antigen specific and compartmentalized to the liver, the site of viral replication.


Activity of Sulbactam-Durlobactam and Comparators Against a National Collection of Carbapenem-Resistant Acinetobacter baumannii Isolates From Greece.
Acinetobacter baumannii is a leading cause of healthcare-associated infections worldwide, due to both its persistence in the hospital setting and ability to acquire high levels of antibiotic resistance. Carbapenem-resistant A. baumannii isolates (CRAB) limit the activity of current antimicrobial regimens and new alternatives or adjuncts to traditional antibiotics are urgently needed. Durlobactam is a novel broad-spectrum inhibitor of serine-type β-lactamases that restores sulbactam (SUL) activity against A. baumannii. The sulbactam-durlobactam (SD) combination has recently completed Phase 3 testing in the global ATTACK trial. The aim of this study is to evaluate the in vitro activity of SD versus comparators against a representative nationwide collection of CRAB isolates. One hundred ninety CRAB isolates were collected from clinical samples of patients hospitalized in 11 hospitals throughout Greece during 2015. In vitro activities of SD and comparators (SUL alone, amikacin, minocycline, imipenem, meropenem, colistin, SD and imipenem combined with SD) were determined by broth microdilution. Durlobactam restored sulbactam activity against the majority of the strains tested, with SD exhibiting the lowest MIC90 (8 μg/ml) relative to the other single comparators tested; 87.9% of the isolates had SD MICs ≤4/4 µg/ml. The most active comparator was colistin (MIC90 = 16 μg/ml). The addition of imipenem further lowered the MIC90 of SD by one two-fold dilution. This study demonstrated the potential utility of SD for the treatment of infections caused by A. baumannii. If its clinical efficacy is confirmed, SD may be an important therapeutic option for CRAB infections.


A systematic review of the clinical features of pneumonia in children aged 5-9 years: Implications for guidelines and research.
Childhood pneumonia presents a large global burden, though most data and guidelines focus on children less than 5 years old. Less information is available about the clinical presentation of pneumonia in children 5-9 years of age. Appropriate diagnostic and treatment algorithms may differ from those applied to younger children. This systematic literature review aimed to identify clinical features of pneumonia in children aged 5-9 years, with a focus on delineation from other age groups and comparison with existing WHO guidance for pneumonia in children less than 5 years old. We searched MEDLINE, EMBASE and PubMed databases for publications that described clinical features of pneumonia in children 5-9 years old, from any country with no date restriction in English. The quality of included studies was evaluated using a modified Effective Public Health Project Practice (EPHPP) tool. Data relating to research context, study type, clinical features of pneumonia and comparisons with children less than 5 years old were extracted. For each clinical feature of pneumonia, we described mean percentage (95% confidence interval) of participants with this finding in terms of aetiology (all cause vs Mycoplasma pneumoniae), and method of diagnosis (radiological vs clinical). We included 15 publications, eight addressing all-cause pneumonia and seven addressing Mycoplasma pneumoniae. Cough and fever were common in children aged 5-9 years with pneumonia. Tachypnoea was documented in around half of patients. Dyspnoea/difficulty breathing and chest indrawing were present in approximately half of all-cause pneumonia cases, with no data on indrawing in the outpatient setting. Chest and abdominal pain were documented in around one third of cases of all-cause pneumonia, based on limited numbers. In addition to markers of pneumonia severity used in children <5 years, pallor has been identified as being associated with poorer outcomes alongside comorbidities and nutritional status. Quality research exploring clinical features of pneumonia, treatment and outcomes in children aged 5-9 years using consistent inclusion criteria, definitions of features and age ranges are urgently needed to better inform practice and guidelines. Based on limited data fever and cough are common in this age group, but tachypnoea cannot be relied on for diagnosis. While waiting for better evidence, broader attention to features such as chest and abdominal pain, the role of chest radiographs for diagnosis in the absence of symptoms such as tachypnoea, and risk factors which may influence patient disposition (chest indrawing, pallor, nutritional status) warrant consideration by clinicians. PROSPERO: CRD42020213837.


Experimental diabetes in lactating sheep: effects of alloxan on plasma insulin, glucose, glucose kinetics and milk characteristics.
After intravenous administration of alloxan (50 mg kg-1 liveweight) to lactating ewes, there were triphasic changes in plasma glucose and insulin. Almost immediately, plasma insulin decreased and hyperglycaemia occurred, then, between c. 5-12 h, insulin increased and ewes became hypoglycaemic. Thereafter, insulin decreased and glucose increased from c. 20 h after alloxan and the diabetic state was established. Changes in glucose production and utilization correlated with changes in plasma glucose. Exogenous insulin was administered from 30 h after alloxan, and it took some 2 weeks to stabilize ewes. During this period, when mild hyperglycaemia persisted, milk yields and feed intakes were decreased but milk fat content was elevated. Once ewes were stabilized, plasma glucose, milk yield, feed intake and milk fat content returned to levels prior to alloxan. These observations are consistent with insulin playing a role in the aetiology of the 'low milk fat syndrome' in the ruminant. It appears that the alloxan-treated, insulin-stabilized ewe would be a useful model for studying the role of insulin during lactation, but it is necessary to allow time for animals to overcome effects of administration of alloxan.