Selective lack of antibody to a component of EB nuclear antigen in patients with chronic active Epstein-Barr virus infection.
The sera of 12 patients with presumed chronic active Epstein-Barr virus (EBV) infection lacked antibody to a component of the Epstein-Barr nuclear antigen (EBNA) complex encoded by the BamHI K fragment of viral DNA. This anomaly, detected in approximately 18% of sera obtained from patients with a diagnosis of "chronic mononucleosis," was more often found in patients with severe disease (approximately 32%) who had objective clinical findings and markedly elevated antibody titers to EBV replicative antigens than in those patients with the "fatigue syndrome" (10%). The lack of antibody to the K nuclear antigen is specific because most of those who did not have antibody to the K antigen made antibody to other latent nuclear (EBNA 2) antigens or nuclear early antigens. Such patients are thus able to lyse immortalized cells, release nuclear products, and present them to the immune system. Three hypotheses are suggested to explain the lack of antibody to the K antigen: a viral mutation, a failure of immune recognition, or lack of in vivo expression of the antigen due to extensive viral replication. Lack of antibody to one component of EBNA may serve as an objective serological marker for certain patients with chronic EBV infection.


New pharmacotherapy options for chemotherapy-induced alimentary mucositis.
Chemotherapy-induced alimentary mucositis is an extremely common condition that is caused by a breakdown of the mucosal barrier. It occurs in between 40 - 100% of cancer patients depending on the treatment regimen. Symptoms typically include pain from oral ulceration, vomiting and diarrhoea. Alimentary mucositis often necessitates chemotherapy reductions or treatment breaks, overall potentially compromising survival outcomes. Consequently, alimentary mucositis creates a burden not only on patients' quality of life but also on healthcare costs. Despite this, currently, there is no clinically effective localised/pharmacological therapy intervention strategy to prevent alimentary mucositis. Over recent years, a number of novel pharmacotherapy agents have been trialed in various preclinical and clinical settings. This critical review will therefore provide an overview of emerging pharmacotherapies for the treatment of alimentary mucositis following chemotherapy with particular emphasis on studies published in the last 2 years. A Pubmed literature search was conducted to identify eligible articles published before 30 November 2013 and each article was reviewed by all authors. All articles were written in English. Currently, there is no clinically effective localised therapeutic intervention strategy to prevent the condition. New emerging areas of research have recently been proposed to play key roles in the development of alimentary mucositis and these areas may provide researchers and clinicians with new research directions. Hopefully this will continue, and evidence-based informed guidelines can be produced to improve clinical practice management of this condition.


Amelioration of sepsis-induced hepatic and ileal injury in rats by the leukotriene receptor blocker montelukast.
Sepsis is a generalized inflammatory response, which involves organ systems remote from the locus of the initial infectious insult, involves the release of cytokines and the subsequent formation of reactive oxygen and nitrogen species. The aim of this study was to investigate the possible protective effect of montelukast, a leukotriene receptor blocker, against oxidative damage in the liver and ileum of septic rats. Sepsis was induced by cecal ligation and puncture method in female Wistar albino rats. Sepsis and sham operated (control) groups received either saline or montelukast (10 mg/kg, ip) immediately after the operation and at 12 h. Twenty-four hours after the surgery, rats were decapitated and malondialdehyde (MDA) content--an index of lipid peroxidation, glutathione (GSH) levels--a key antioxidant, myeloperoxidase (MPO) activity--an index of neutrophil infiltration, and collagen contents were determined in the liver and ileum. Formation of reactive oxygen species in liver and ileal tissue samples was monitored by using chemiluminescence (CL) technique with luminol and lucigenin probes. Both tissues were also analyzed histologically. Serum lactate dehydrogenase (LDH) and tumor necrosis factor-alpha (TNF-alpha) level were assessed in trunk blood. Sepsis resulted in decreased GSH levels, and increased MDA levels, MPO activity, CL levels and collagen contents in both the liver and the ileum (P < 0.05-P < 0.001) indicating the presence of the oxidative damage. Similarly, serum TNF-alpha and LDH were elevated in the sepsis group as compared to control group. On the other hand, montelukast treatment reversed all these biochemical indices, as well as histopathological alterations, which were induced by sepsis. Findings of the present study suggest that montelukast possesses an anti-inflammatory effect on sepsis-induced hepatic and intestinal damage and protects against oxidative injury by a neutrophil-dependent mechanism.


Silicotungstate, a Potential Electron Transporting Layer for Low-Temperature Perovskite Solar Cells.
Thin films of a heteropolytungstate, lithium silicotungstate (Li4SiW12O40, termed Li-ST), prepared by a solution process at low temperature, were successfully applied as electron transporting layer (ETL) of planar-type perovskite solar cells (PSCs). Dense and uniform Li-ST films were prepared on FTO glass by depositing a thin Li-ST buffer layer, followed by coating of a main Li-ST layer. The film thickness was controlled by varying the number of coating cycles, consisting of spin-coating and thermal treatment at 150 °C. In particular, by employing 60 nm-thick Li-ST layer obtained by two cycles of coating, the fabricated CH3NH3PbI3 PSC device demonstrates the photovoltaic conversion efficiency (PCE) of 14.26% with JSC of 22.16 mA cm-2, VOC of 0.993 mV and FF of 64.81%. The obtained PCE is significantly higher than that of the PSC employing a TiO2 layer processed at the same temperature (PCE = 12.27%). Spectroscopic analyses by time-resolved photoluminescence and pulsed light-induced transient measurement of photocurrent indicate that the Li-ST layer collects electrons from CH3NH3PbI3 more efficiently and also exhibits longer electron lifetime than the TiO2 layer thermally treated at 150 °C. Thus, Li-ST is considered to be a promising ETL material that can be applied for the fabrication of flexible PSC devices.


Spironolactone differently influences remodeling of the left ventricle and aorta in L-NAME-induced hypertension.
Aldosterone receptor antagonist, spironolactone, has been shown to prevent remodeling of the heart in several models of left ventricular hypertrophy. The aim of the present study was to determine whether the treatment with spironolactone can prevent hypertension, reduction of tissue nitric oxide synthase activity and left ventricular (LV) and aortic remodeling in N(G)-nitro-L-arginine methyl ester (L-NAME)-induced hypertension. Four groups of rats were investigated: control, spironolactone (200 mg/kg), L-NAME (40 mg/kg) and L-NAME + spironolactone (in corresponding dosage). Animals were studied after 5 weeks of treatment. The decrease of NO-synthase activity in the LV and kidney was associated with the development of hypertension and LV hypertrophy, with increased DNA concentration in the LV, and remodeling of the aorta in the L-NAME group. Spironolactone prevented the inhibition of NO-synthase activity in the LV and kidney and partially attenuated hypertension and LVH development and the increase in DNA concentration. However, remodeling of the aorta was not prevented by spironolactone treatment. We conclude that the aldosterone receptor antagonist spironolactone improved nitric oxide production and partially prevented hypertension and LVH development without preventing hypertrophy of the aorta in NO-deficient hypertension. The reactive growth of the heart and aorta seems to be controlled by different mechanisms in L-NAME-induced hypertension.


Intracerebral arteriovenous malformation fed by the anterior ethmoidal artery: case report.
The authors report a case of intracerebral arteriovenous malformation fed by the anterior ethmoidal artery. The nidus was located in the brain parenchyma, although its feeder was a dural artery. The etiology of this rare condition is discussed.


Lemon myrtle extract inhibits lactate production by Streptococcus mutans.
Backhousia citriodora (lemon myrtle) extract has been found to inhibit glucansucrase activity, which plays an important role in biofilm formation by Streptococcus mutans. In addition to glucansucrase, various virulence factors in S. mutans are involved in the initiation of caries. Lactate produced by S. mutans demineralizes the tooth enamel. This study investigated whether lemon myrtle extract can inhibit S. mutans lactate production. Lemon myrtle extract reduced the glycolytic pH drop in S. mutans culture and inhibited lactate production by at least 46%. Ellagic acid, quercetin, hesperetin, and myricetin, major polyphenols in lemon myrtle, reduced the glycolytic pH drop and lactate production, but not lactate dehydrogenase activity. Furthermore, these polyphenols reduced the viable S. mutans cell count. Thus, lemon myrtle extracts may inhibit S. mutans-mediated acidification of the oral cavity, thereby preventing dental caries and tooth decay.


Effects of verapamil in ventricular tachycardias. An experimental and clinical study.
To determine the effect of verapamil in ventricular tachycardias, we performed an experimental and clinical study. Experimental ventricular tachycardias (VT) were produced in dog hearts with minute aconitine crystals introduced into the periphery of a left ventricular area, damaged by intramural injection of 1.0-1.5 ml phenol. The response of these tachycardias to 0.2 mg/kg verapamil was analyzed. Verapamil was infused into the superior vena cava over 15-20 min. Leads II, aVL, intraventricular right and left unipolar records, as well as one of the superior vena cava, were registered under control conditions, in the presence of VT, and after application of verapamil. Recordings were obtained at constant intervals, waiting for the recovery of sinus rhythm (SR) and the posterior reappearance of tachycardia. Experiments were performed for 6 to 8 h under continuous infusion of Hartmann's solution. Throughout these periods, variations in systemic systolic pressure were recorded. From 75 animals submitted to this treatment, 30 (40%) recovered transiently the SR, whereas the drug exerted no antiarrhythmic effect in 19 (25%), and arterial systolic pressure fell importantly in 10 (13%) animals. In two more groups, of 15 dogs each, the VT response to verapamil was compared with the response to lidocaine and flecainide. Endovenous verapamil (5-10 mg) was administered to 10 patients, coursing with VT and having a structurally normal heart, after this arrhythmia was induced by electrical stimulation. The response to verapamil was satisfactory in nine patients (90%), in which VT originated in the septal and apical regions of the left ventricle. Verapamil seems to be effective in experimental and clinical ventricular tachycardias related to calcium-dependent potentials, in which the sustaining mechanism could either be triggered activity or reentry.


Stem Cells and Their Derivatives: An Implication for the Regeneration of Nonunion Fractures.
Despite advances in biomedical research, fracture nonunion rates have remained stable throughout the years. Long-bone fractures have a high likelihood of nonunion, but the specific biological pathways involved in this severe consequence are unknown. Fractures often heal in an organized sequence, including the production of a hematoma and an early stage of inflammation, the development of a soft callus and hard callus, and eventually the stage of bone remodeling. Deficient healing can result in a persistent bone defect with instability, discomfort, and loss of function. In the treatment of nonunions, mesenchymal stem cells (MSCs) prove to be a promising and safe alternative to the standard therapeutic strategies. Moreover, novel scaffolds are being created in order to use a synergistic biomimetic technique to rapidly generate bone tissue. MSCs respond to acellular biomimetic matrices by regenerating bone. Extracellular vesicles (EVs) derived from MSCs have recently gained interest in the field of musculoskeletal regeneration. Although many of these techniques and technologies are still in the preclinical stage and have not yet been approved for use in humans, novel approaches to accelerate bone healing via MSCs and/or MSC derivatives have the potential to reduce the physical, economic, and social burdens associated with nonhealing fractures and bone defects. In this review, we focus on providing an up-to-date summary of recent scientific studies dealing with the treatment of nonunion fractures in clinical and preclinical settings employing MSC-based therapeutic techniques.


Thresholds for self-motion perception in roll without and with visual fixation target--the visualvestibular interaction effect.
The purpose of this study was to establish the selfmotion perception threshold, in roll, in the visualvestibular interaction (VVI) state, creating an oculogyral illusion, and to compare this threshold to the self-motion perception threshold in darkness. A further aim was to investigate the dynamics of the threshold at a low frequency range (0.1-1 Hz) of sinusoidal rotation. Seven healthy subjects were tested. A motion platform was used to generate motion. Single cycles of sinusoidal acceleration at four frequencies (0.1, 0.2, 0.5 and 1 Hz) were used as motion stimuli. To avoid otolith stimulation, subjects were rotated about a vertical axis in supine position. To evoke an oculogyral illusion subjects were instructed to fixate their gaze on a cross-shaped object aligned with their head, which rotated with them. The results show a lowering of the self-motion perception threshold in the VVI state, significant for the frequencies 0.1 and 0.2 Hz (p<0.05). In all the subjects, visual fixation on the cross evoked an oculogyral illusion. The threshold in both tested conditions was frequency dependent: it decreased with increasing frequency values. However, this effect was consistently stronger in darkness across all frequencies (p<0.05). In conclusion, the application of sinusoidal rotation during roll at low frequencies in the VVI condition evokes oculogyral illusion. This interaction lowers the self-motion perception threshold compared to that measured during rotation in darkness. This testing method could be of practical benefit in clinical application for revealing brain dysfunction involving integrative mechanisms of perception.


Engineering Proteins for Thermostability with iRDP Web Server.
Engineering protein molecules with desired structure and biological functions has been an elusive goal. Development of industrially viable proteins with improved properties such as stability, catalytic activity and altered specificity by modifying the structure of an existing protein has widely been targeted through rational protein engineering. Although a range of factors contributing to thermal stability have been identified and widely researched, the in silico implementation of these as strategies directed towards enhancement of protein stability has not yet been explored extensively. A wide range of structural analysis tools is currently available for in silico protein engineering. However these tools concentrate on only a limited number of factors or individual protein structures, resulting in cumbersome and time-consuming analysis. The iRDP web server presented here provides a unified platform comprising of iCAPS, iStability and iMutants modules. Each module addresses different facets of effective rational engineering of proteins aiming towards enhanced stability. While iCAPS aids in selection of target protein based on factors contributing to structural stability, iStability uniquely offers in silico implementation of known thermostabilization strategies in proteins for identification and stability prediction of potential stabilizing mutation sites. iMutants aims to assess mutants based on changes in local interaction network and degree of residue conservation at the mutation sites. Each module was validated using an extensively diverse dataset. The server is freely accessible at http://irdp.ncl.res.in and has no login requirements.


Multiple mechanisms for TRAF3-mediated regulation of the T cell costimulatory receptor GITR.
Tumor necrosis factor receptor (TNFR)-associated factor 3 (TRAF3) plays context-specific roles in multiple receptor-mediated signaling pathways in different cell types. Mice lacking TRAF3 in T cells display defective T-cell-mediated immune responses to immunization and infection and demonstrate defective early signaling via the TCR complex. However, the role of TRAF3 in the function of GITR/TNFRSF18, an important costimulatory member of the TNFR superfamily, is unclear. Here we investigated the impact of T cell TRAF3 status on both GITR expression and activation of specific kinases in the GITR signaling pathway in T cells. Our results indicate that TRAF3 negatively regulates GITR functions in several ways. First, expression of GITR protein was elevated in TRAF3-deficient T cells, resulting from both transcriptional and posttranslational regulation that led to greater GITR transcript levels, as well as enhanced GITR protein stability. TRAF3 associated with T cell GITR in a manner dependent upon GITR ligation. TRAF3 also inhibited several events of the GITR mediated early signaling cascade, in a manner independent of recruitment of phosphatases, a mechanism by which TRAF3 inhibits signaling through several other cytokine receptors. These results add new information to our understanding of GITR signaling and function in T cells, which is relevant to the potential use of GITR to enhance immune therapies.


A quantum chemical study of H2S2: Intramolecular torsional mode and intermolecular interactions with rare gases.
The structural and energetic properties of the H(2)S(2) molecule have been studied using density functional theory, second-order Moller-Plesset method, and coupled cluster theory with several basis sets. In order to extend previous work on intra- and intermolecular dynamics of the chirality changing modes for H(2)O(2) and its derivatives, our focus has been on the torsion around the S-S bond, along with an extensive characterization of the intermolecular potentials of H(2)S(2) with the rare gases (He, Ne, Ar, and Kr). Use is made of previously defined coordinates and expansion formulas for the potentials which allow for a faithful representation of geometrical and symmetry properties of these systems that involve the interaction of an atom with a floppy molecule. The potential energy surfaces obtained in this work are useful for classical and quantum mechanical simulations of molecular collisions responsible for chirality changing processes of possible interest in the modeling of prebiotic phenomena.


Molecular epidemiology of Mycobacterium tuberculosis complex in the Volta Region of Ghana.
Available molecular epidemiological data from recent studies suggest significant genetic variation between the different lineages of Mycobacterium tuberculosis complex (MTBC) and the MTBC lineages might have adapted to different human populations. This study sought to determine the population structure of clinical MTBC isolates from the Volta Region of Ghana. The MTBC isolates obtained from collected sputum samples were identified by PCR detecting of IS6110 and genotyped using spoligotyping. Non-tuberculous mycobacterial isolates were characterized by amplification of the heat shock protein 65 (hsp65) gene and sequencing. The drug susceptibility profiles of the MTBCs determined using GenoType MTBDRplus. One hundred and seventeen (117, 93.6%) out of 125 mycobacterial positive isolates were characterized as members of the MTBC of which M. tuberculosis sensu stricto (MTBss) and M. africanum (MAF) were respectively 94 (80.3%) and 23 (19.7%). In all, 39 distinct spoligotype patterns were obtained; 26 for MTBss and 13 for MAF lineages. Spoligotyping identified 89 (76%) Lineage 4, 16 (13.6%) Lineage 5, 7 (6.0%) Lineage 6, 3 (2.6%) Lineage 2, 1(0.9%) Lineage 3 and 1 (0.9%) Lineage 1. Among the Lineage 4 isolates, 62/89 (69.7%) belonged to Cameroon sub-lineage, 13 (14.7%) Ghana, 8 (9.0%) Haarlem, 2 (2.2%) LAM, 1 (1.1%) Uganda I, 1 (1.1%) X and the remaining two (2.2%) were orphan. Significant localization of MAF was found within the Ho municipality (n = 13, 29.5%) compared to the more cosmopolitan Ketu-South/Aflao (n = 3, 8.3%) (p-value = 0.017). Eight (8) non-tuberculous mycobacteria were characterized as M. abscessus (7) and M. fortuitum (1). We confirmed the importance of M. africanum lineages as a cause of TB in the Volta region of Ghana.


Epitope-driven DNA vaccine design employing immunoinformatics against B-cell lymphoma: a biotech's challenge.
DNA vaccination has been widely explored to develop new, alternative and efficient vaccines for cancer immunotherapy. DNA vaccines offer several benefits such as specific targeting, use of multiple genes to enhance immunity and reduced risk compared to conventional vaccines. Rapid developments in molecular biology and immunoinformatics enable rational design approaches. These technologies allow construction of DNA vaccines encoding selected tumor antigens together with molecules to direct and amplify the desired effector pathways, as well as highly targeted vaccines aimed at specific epitopes. Reliable predictions of immunogenic T cell epitope peptides are crucial for rational vaccine design and represent a key problem in immunoinformatics. Computational approaches have been developed to facilitate the process of epitope detection and show potential applications to the immunotherapeutic treatment of cancer. In this review a number of different epitope prediction methods are briefly illustrated and effective use of these resources to support experimental studies is described. Epitope-driven vaccine design employs these bioinformatics algorithms to identify potential targets of vaccines against cancer. In this paper the selection of T cell epitopes to develop epitope-based vaccines, the need for CD4(+) T cell help for improved vaccines and the assessment of vaccine performance against tumor are reviewed. We focused on two applications, namely prediction of novel T cell epitopes and epitope enhancement by sequence modification, and combined rationale design with bioinformatics for creation of new synthetic mini-genes. This review describes the development of epitope-based DNA vaccines and their antitumor effects in preclinical research against B-cell lymphoma, corroborating the usefulness of this platform as a potential tool for cancer therapy. Achievements in the field of DNA vaccines allow to overcome hurdles to clinical translation. In a scenario where the vaccine industry is rapidly changing from a mostly empirical approach to a rational design approach, these new technologies promise to discover and develop high-value vaccines, creating a new opportunity for future markets.


Reduction of airborne microorganisms by filtering recycled air in a chick hatcher.
An experimental chick hatcher designed to filter recycled ventilation air was tested for its effectiveness in reducing the number of viable airborne microorganisms. Chicks in a filtered hatcher and a control hatcher (no filter) were artificially contaminated with Serratia marcescens as ventilation air was recycled in the hatchers for twelve hours. The number of viable S. marcescens particles in the filtered air of the conditioning chamber was less than detectable. The number of viable airborne S. marcescens particles in the hatching chamber of the filtered hatcher indicated a reduction of greater than 90 percent over the number in the unfiltered hatcher. The filter was effective in reducing the number of airborne particles carrying viable S. marcescens organisms in the hatcher.


Illicit drug use and work in a model therapeutic workplace.
The link between illicit drug use and impaired employee performance in the workplace has been assumed, but the relation has not been demonstrated clearly in research. This study was an evaluation of the relations between cocaine and opiate use, attendance, and performance in a job skills training program in a population with high rates of drug use. Out-of-treatment injection drug users (N = 42) attended a model therapeutic workplace where they could earn a maximum pay of around $10 per hour, 4 h every weekday, for 30 weeks. At the workplace, participants could complete practice trials on computer-based typing and keypad training programs. Participants were asked to provide urine samples thrice weekly, which were tested for opiates and cocaine. Participants worked for more hours on a program that resulted in a flat hourly wage when their urine was negative for opiates and cocaine than when their urine was opiate and cocaine positive. Attendance was positively associated with opiate-negative samples during the study. When participants attended the workplace, however, their performance was not related to drug use. Participants completed the same number of practice trials, performed at the same accuracy, and typed at the same speed when they were positive and negative for cocaine and opiates. Contrary to common expectations, this study failed to show that the use of opiates or cocaine affected in-training performance, even though opiate and cocaine use predicted reduced attendance under some circumstances.


Pediatric and adolescent applications of the Taylor Spatial Frame.
Limb deformity can occur in the pediatric and adolescent populations from multiple etiologies: congenital, traumatic, posttraumatic sequelae, oncologic, and infection. Correcting these deformities is important for many reasons. Ilizarov popularized external fixation to accomplish this task. Taylor expanded on this by designing an external fixator in 1994 with 6 telescoping struts that can be sequentially manipulated to achieve multiaxial correction of deformity without the need for hinges or operative frame alterations. This frame can be used to correct deformities in children and has shown good anatomic correction with minimal morbidity. The nature of the construct and length of treatment affects psychosocial factors that the surgeon and family must be aware of prior to treatment. An understanding of applications of the Taylor Spatial Frame gives orthopedic surgeons an extra tool to correct simple and complex deformities in pediatric and adolescent patients.


On-chip magnetic separation of superparamagnetic beads for integrated molecular analysis.
We have demonstrated a postprocessed complementary metal oxide semiconductor (CMOS) integrated circuit (IC) capable of on-chip magnetic separation, i.e., removing via magnetic forces the nonspecifically bound magnetic beads from the detection area on the surface of the chip. Initially, 4.5 mum wide superparamagnetic beads sedimenting out of solution due to gravity were attracted to the detection area by a magnetic concentration force generated by flowing current through a conductor embedded in the IC. After sedimentation, the magnetic beads that did not bind strongly to the functionalized surface of the IC through a specific biochemical complex were removed by a magnetic separation force generated by flowing current through another conductor placed laterally to the detection area. As the spherical bead pivoted on the surface of the chip, the lateral magnetic force was further amplified by mechanical leveraging, and 50 mA of current flowing through the separation conductor placed 18 mum away from the bead resulted in 7.5 pN of tensile force on the biomolecular tether immobilizing the bead. This force proved high enough to break nonspecific interactions while leaving specific antibody-antigen bonds intact. A sandwich capture immunoassay on purified human immunoglobulin G showed strong correlation with a control enzyme linked immunosorbent assay and a detection limit of 10 ngml or 70 pM. The beads bound to the detection area after on-chip magnetic separation were detected optically. To implement a fully integrated molecular diagnostics platform, the on-chip magnetic separation functionality presented in this work can be readily combine with state-of-the art CMOS-based magnetic bead detection technology.


Isolation and characterization of neurokinin A, neurokinin A(3-10) and neurokinin A(4-10) from a neutral water extract of a metastatic ileal carcinoid tumour.
A metastasis to the right liver lobe of an argyrophil/argentaffin midgut carcinoid tumour in a patient with the classical carcinoid syndrome was examined for the presence of tachykinins other than substance P, using a specific antiserum. The extract was initially purified using SepPak cartridges, and subsequently subjected to cation-exchange chromatography on SP Sephadex C-25 which separated the immunoreactive material into two main components (components I and II). Both were further purified by anion-exchange chromatography on DEAE-Sephadex A-25, and by reverse-phase fast protein liquid chromatography. Component II was identified as neurokinin A by its immunochemical and chromatographic properties and amino acid sequence analysis. Component I consisted of two molecular forms which were identified as neurokinin A(3-10) and neurokinin A(4-10) by amino acid sequence analysis. The tumour tissue contained only small amounts of the eledoisin-like peptide that has earlier been demonstrated in mammalian tissues. Although this component behaved like the nonmammalian peptide eledoisin on reverse-phase HPLC and on reverse-phase ion-pair chromatography, eledoisin-specific antiserum E2 indicated that eledoisin-like peptide is not identical to eledoisin. Neurokinin A in carcinoid tumours has an N-terminal heterogeneity; this multiplicity constitutes a further support for the hypothesis that carcinoid tumours produce a number of tachykinins which may be present in different relative amounts in individual patients and may contribute to the individual differences in symptomatology.


Three new diterpenes from the marine soft coral Lobophytum crassum.
Three new terpenoid metabolites (1-3) were isolated from the CH2Cl2 extract of the soft coral Lobophytum crassum together with the eudesmane derivative 4 and the known cembrane (2S,7S,8S)-sarcophytoxide (5). Compound 1 is a new cembrane-based diterpene with an C7-C8-epoxide and a methyl ester functionality at C-16. (3E,5Z)- (2) and (3Z,5E)-2-methyl-6-(4a'-methyl-8'-methylene-trans-perhydr onaphthalen-2'-yl)hepta-3,5-dien-2-ol (3) represent two new carbon-carbon double bond isomers of 4, which has the 3E,5E-configuration. The structures of 1-5 were established by interpretation of their spectroscopic data, mainly 1D and 2D NMR and MS. Biological activity evaluation of compounds 1 and 5 and the crude extracts was carried out using agar diffusion assays toward microbial targets and ELISA assays for investigating the inhibition of HIV-1 reverse transcriptase and p56lck tyrosine kinase.


Development of a sexual function questionnaire for clinical trials of female sexual dysfunction.
To better evaluate efficacy in clinical trials of drugs as potential treatments for female sexual dysfunctions (FSD), a brief, multidimensional measure of female sexual function was developed. Data from semistructured interviews with 82 women with or without FSD, aged 19-65 years, generated a pool of 61 items that addressed aspects of female sexual function. On review by a panel, individual items were selected for face validity and clinical relevance. Thirty-one items were used as a sexual function questionnaire (SFQ-V1) in two multicenter, phase II clinical trials totaling 781 women with FSD. Normative data were generated from a sample of 201 women without FSD. Factor analysis produced seven domains of female sexual function: desire, physical arousal-sensation, physical arousal-lubrication, enjoyment, orgasm, pain, and partner relationship. The internal consistency of the domains ranged from 0.65 to 0.91, and test-retest reliability ranged from 0.21 to 0.71 for Cohen's weighted kappa and 0.42 to 0.78 for Pearson's correlation coefficient. There was a significant difference between the baseline mean SFQ domain scores of patients with FSD compared with those of women without FSD (p < 0.0001). End-of-study SFQ scores were significantly different for women who reported improvement vs. women who reported no improvement (p < 0.001). The SFQ produced seven domains of female sexual function with excellent internal consistency, moderate to good reliability, excellent discriminant validity, and sensitivity. The results suggest that the SFQ may be a valuable new tool for evaluating and diagnosing subsets of FSD and, ultimately, for evaluating treatments of these disorders.


FUGE: an analytical ultracentrifuge simulation for teaching purposes.
An IBM-compatible microcomputer program for teaching purposes is described which stimulates the operation of a sedimentation velocity determination of a protein in an analytical ultracentrifuge using schlieren optics. The program operates in speeded-up time and simulates the major procedures which would need to be carried out to operate such an instrument. The position of the sedimenting boundary can be observed at any time during the run, and up to six 'photographs' can be recorded for subsequent analysis. Calculation of sedimentation coefficient, diffusion coefficient and mol. wt can be made from a dot-matrix printout. Ten representative proteins are stored within the program, but provision exists for user-supplied data.


Sensitizing immune unresponsive colorectal cancers to immune checkpoint inhibitors through MAVS overexpression.
The majority of colorectal carcinomas (CRCs) are insensitive to programmed death protein-1/programmed death-ligand 1 (anti-PD-1/PD-L1) immune checkpoint inhibitor (ICI) antibodies. While there are many causes for ICI insensitivity, recent studies suggest that suppression of innate immune gene expression in tumor cells could be a root cause of this insensitivity and an important factor in the evolution of tumor immunosuppression. We first assessed the reduction of mitochondrial antiviral signaling gene (MAVS) and related RIG-I pathway gene expression in several patient RNA expression datasets. We then engineered MAVS expressing tumor cells and tested their ability to elicit innate and adaptive anti-tumor immunity using both in vitro and in vivo approaches, which we then confirmed using MAVS expressing viral vectors. Finally, we observed that MAVS stimulated PD-L1 expression in multiple cell types and then assessed the combination of PD-L1 ICI antibodies with MAVS tumor expression in vivo. MAVS was significantly downregulated in CRCs, but its re-expression could stimulate broad cellular interferon-related responses, in both murine and patient-derived CRCs. In vivo, local MAVS expression elicited significant anti-tumor responses in both immune-sensitive and insensitive CRC models, through the stimulation of an interferon responsive axis that provoked tumor antigen-specific adaptive immunity. Critically, we found that tumor-intrinsic MAVS expression triggered systemic adaptive immune responses that enabled abscopal CD8 +T cell cytotoxicity against distant CRCs. As MAVS also induced PD-L1 expression, we further found synergistic anti-tumor responses in combination with anti-PD-L1 ICIs. These data demonstrate that intratumoral MAVS expression results in local and systemic tumor antigen-specific T cell responses, which could be combined with PD-L1 ICI to permit effective anti-tumor immunotherapy in ICI resistant cancers.


Preoperative assessment of intestinal endometriosis: A comparison of transvaginal sonography with water-contrast in the rectum, transrectal sonography, and barium enema.
To evaluate the accuracy of Transrectal Sonography (TRS) and a new technique, Transvaginal Sonography with Water-Contrast in the Rectum (RWC-TVS), in the diagnosis of rectosigmoid endometriosis, and the accuracy of Barium Enema (BE) and RWC-TVS in the detection of intestinal stenosis due to endometriosis. In a prospective study, we compared the findings of TRS and RWC-TVS performed before surgery with the operative and pathologic findings in 61 consecutive patients who underwent laparoscopy or laparotomy for suspected rectosigmoid endometriosis. The accuracy of BE and RWC-TVS in the detection of intestinal stenosis was evaluated comparing the radiologic and ultrasonographic results with the macroscopic findings at surgery and pathology. RWC-TVS diagnosed rectosigmoid endometriosis with the same accuracy of TRS and was equally efficient as BE in the detection of a significant intestinal lumen stenosis. For the diagnosis of rectosigmoid endometriosis the sensitivity, specificity, positive and negative predictive values of TRS and RWC-TVS were 88.2% and 96%, 80%, and 90%, 95.7%, and 98%, and 57.1% and 81.8%, respectively. For the detection of intestinal stenosis the sensitivity, specificity, positive and negative predictive values of BE and RWC-TVS were 93.7% and 87.5%, 94.2% and 91.4%, 88.2% and 82.3%, and 97% and 94.1%, respectively. RWC-TVS is a new, simple technique for a single-step and accurate preoperative assessment of rectosigmoid endometriosis.


Semantic profiles in mild cognitive impairment associated with Alzheimer's and Parkinson's diseases.
The temporal and the prefrontal cortices have different roles in semantic information processing: the temporal lobe is where knowledge is stored (Graham and Hodges, 1997), whereas the prefrontal cortex is more specifically involved in executive aspects of semantic processing. Relatively little is known about the semantic profiles of mild cognitive impairment (MCI) in Alzheimer's disease (AD) and Parkinson's disease (PD). This observational study investigated naming and semantic questionnaire performances in three groups of subjects: 10 patients with the amnestic-type MCI prodrome of AD (aMCI), 10 patients with early-stage executive-type MCI in PD (MCI-PD), and 10 normal subjects. The MCI-PD subjects demonstrated inferior performances on a semantic questionnaire, whereas the aMCI group displayed modest difficulties in a naming task. These differences may be explained by topographical differences in pathological involvement. Since the frontal areas are more functionally impaired in PD, we hypothesize that the semantic deficit may be a consequence of a deficiency in control of semantic processing. On the other hand, the semantic deficit in aMCI may be related to a lexical-semantic storage dysfunction resulting from pathological involvement of the temporal lobe.


Trial of a Novel Intervention to Improve Multiple Food Hygiene Behaviors in Nepal.
AbstractIn this study, we report on the results of a trial of an intervention to improve five food hygiene behaviors among mothers of young children in rural Nepal. This novel intervention targeted five behaviors; cleanliness of serving utensils, handwashing with soap before feeding, proper storage of cooked food, and thorough reheating and water treatment. Based on formative research and a creative process using the Behavior-Centered Design approach, an innovative intervention package was designed and delivered over a period of 3 months. The intervention activities included local rallies, games, rewards, storytelling, drama, competitions linking with emotional drivers of behavior, and "kitchen makeovers" to disrupt behavior settings. The effect of the package on behavior was evaluated via a cluster-randomized before-after study in four villages with four villages serving as controls. The primary outcome was the difference in the mean cluster level proportions of mothers directly observed practicing all five food hygiene behaviors. The five targeted food hygiene behaviors were rare at baseline (composite performance of all five behaviors in intervention 1% [standard deviation (SD) = 2%] and in control groups 2% [SD = 2%]). Six weeks after the intervention, the target behaviors were more common in the intervention than in the control group (43% [SD = 14%] versus 2% [SD = 2%], P = 0.02) during follow-up. The intervention appeared to be equally effective in improving all five behaviors in all intervention clusters. This study shows that a theory-driven, systematic approach employing emotional motivators and modifying behavior settings was capable of substantially improving multiple food hygiene behaviors in Nepal.


Extended right hepatectomy for hilar bile duct carcinoma using the modified liver hanging maneuver.
To achieve complete extended right hepatectomy or trisectionectomy for a bismuth type IV hilar bile duct carcinoma, we propose the application of Belghiti's liver hanging maneuver (LHM) using a small nasogastric tube. This small nasogastric tube was placed in the cut plane: the top of the tube was placed between the hepatic veins. The tube was placed along the border between the left lateral sector and Spiegel's caudate lobe and the bottom of the tube was placed at the left side of the umbilical Glissonian pedicle. Hepatic parenchyma was transected using a vascular sealing device. Hepatic transection was always targeted to the tube and, eventually, a cut line of left hepatic ducts remained. We report the case of a 76-year-old female and an 83-year-old female with widely extended hilar bile duct carcinomas showing Bismuth type IV. Applying the modified LHM for extended right hepatectomy, the cut planes were easily and adequately obtained in patients with hilar bile duct carcinoma.


Cell loss from three established lines of the C3H mouse mammary tumor: a comparison of the 125I-UdR and the 3H-TdR-autoradiographic methods.
The 125I-UdR method for measuring cell loss from solid tumors has been reevaluated. The rate of tumor cell loss from three established lines (S 102F, S102S and Slow) of the C3H mouse mammary tumor was determined by the 125I-UdR method and the results were compared to the estimates for cell loss as determined by the combined approach of cellular 3H-TdR autoradiography and volumetric growth-rate determinations. This detailed comparison shows that the two methods complement each other but cannot substitute for one another because they give different quantitative information. The combined approach measures the flow of viable cells, as determined morphologically, from the proliferating compartment to the quiescent comparment, the quiescent compartment out of the tumor, etc., but does not evaluate the flow of degenerate cells or acellular (necrotic) debris. In contrast, the 125I-UdR method indicates the net flow of intact cells and/or dead cells as well as debris from the tumor as the 125I-labeled material is lost from the tumor, but gives limited internal information. thus, depending on the specific experiment, an investigator could choose one or the other of the methods to answer the question. Perhaps both would be desirable at times; however, in most cases, one could not substitute one method for the other. The data from the Slow tumors also indicate that in certain tumors, the quantitative information from the 125I-UdR method may be quite limited, i.e. the confidence limits within an experiment as well as the replication error between experiments may be high.


X-ray Crystallographic Structure of a Compact Dodecamer from a Peptide Derived from Aβ16-36.
The assembly of the β-amyloid peptide, Aβ, into soluble oligomers is associated with neurodegeneration in Alzheimer's disease. The Aβ oligomers are thought to be composed of β-hairpins. Here, the effect of shifting the residue pairing of the β-hairpins on the structures of the oligomers that form is explored through X-ray crystallography. Three residue pairings were investigated using constrained macrocyclic β-hairpins in which Aβ30-36 is juxtaposed with Aβ17-23, Aβ16-22, and Aβ15-21. The Aβ16-22-Aβ30-36 pairing forms a compact ball-shaped dodecamer composed of fused triangular trimers. This dodecamer may help explain the structures of the trimers and dodecamers formed by full-length Aβ.


Evaluation of inter- and intramolecular primary structure homologies of interferons by a Monte Carlo method.
Using Sellers TT algorithm, primary structure repeats have been described for interferon (IFN)-alpha, -beta 1, and gamma. To reevaluate these results and to extend them to IFN-beta 2 (interleukin-6), a modified algorithm was developed that uses a metric to define the "best" partial homology of two peptide sequences and to compare it to those detected in random permutations of the peptide. Using this approach, the known structural homologies of IFN-alpha with IFN-beta 1 and of human (Hu) IFN-gamma with murine (Mu) IFN-gamma were identified correctly. However, the primary structure repeats in the amino acid sequences of IFN-alpha, -beta 1, and -gamma turned out to be no better than those detectable in random permutations of these sequences. These results were confirmed using a different, nonlinear metric. A previously used approach to demonstrate significance was shown to produce false-positive results. No significant primary structure homologies were detected among IFN-beta 1, -beta 2, and -gamma. In contrast to the amino acid sequence analysis, the DNA sequence of HuIFN-beta 1 contained a significant repeat that had no significant counterpart in MuIFN-beta or in IFN-alpha. In conclusion, some previously reported results obtained with Sellers TT algorithm on amino acid sequences are easily explained as random similarities, and it is therefore strongly recommended that a method like ours should be used to control significance.


Carvacrol prevents impairments in motor and neurochemical parameters in a model of progressive parkinsonism induced by reserpine.
Parkinson's disease (PD) is a neurodegenerative disease characterized by a progressive degeneration of dopaminergic neurons in the substantia nigra pars compact (SNpc), with consequent depletion of dopamine in the striatum, which gives rise to the characteristic motor symptoms of PD. Although its etiology is unknown, several studies have suggested that oxidative stress plays a critical function in the pathophysiology of PD, and antioxidant agents could be helpful to slown down the dopaminergic neurodegeneration. Carvacrol (CA) is a phenolic monoterpene found in essential oils of many aromatic plants that presents antioxidant and neuroprotective effects. This study aimed to assess the effect of CA in a reserpine (RES)-induced rat model of PD. Male Wistar rats received 15 s.c. injections of 0.1 mg/kg RES or vehicle, every other day, concomitantly to daily i.p. injections of CA (12.5 or 25 mg/kg) or vehicle. Across the treatment, the animals were submitted to behavioral evaluation in the catalepsy test (performed daily), open field test (7th day) and assessment of vacuous chewing movements (12th, 20th and 30th days). Upon completion of behavioral tests, rats were perfused and their brains underwent tyrosine hydroxylase (TH) immunohistochemical analysis. Our results showed that CA (12.5 e 25 mg/kg) prevented the increase in catalepsy behavior and number of vacuous chewing movements, but failed to revert the decreased open-field locomotor activity induced by RES. In addition, CA in both doses prevented the decrease in TH immunostaining induced by RES in the SNpc and dorsal striatum. Taken together, our results suggest that CA shows a protective effect in a rat model of PD, preventing motor and neurochemical impairments induced by RES. Thus, the use of CA as a promising new strategy for the prevention and/or treatment of PD may be considered.


Light microscopic and color television image analysis of the development of staining on chlorhexidine-treated surfaces.
Tooth staining with the use of chlorhexidine preparations is the major problem of long term application. Evidence suggests that the staining arises from a cationic/anionic interacation of chlorhexidine with components of certain dietary materials. The purpose of this in vitro study was to compare visually the development of tea and coffee staining on acrylic and tooth specimens treated with chlorhexidine and to follow the development of tea staining on perspex by light microscopy and color television image analysis. All specimens were maintained in their respective beverage for 5 days with test specimens removed three times a day and placed for 2 minutes in an 0.2% chlorhexidine solution. Both test tooth and acrylic specimens showed comparably and markedly increased staining by the beverages compared with control specimens. Color television image analysis of test specimens demonstrated more marked and rapid development of tea staining when studied on a daily basis. Microscopic examination revealed the staining to be made up of small particles of material which increased in size and coalesced with time. Again, marked differences were apparent in the stain on test and control specimens. The results of this in vitro method provided further evidence for a dietary aetiology to chlorhexidine staining and were consistent with clinical findings. Such a method may be useful to assess staining arising from the use of other anti-plaque agents.


Comparison of RT-PCR, RT-nested PCRs, and real-time PCR for diagnosis of severe fever with thrombocytopenia syndrome: a prospective study.
We designed a highly sensitive reverse transcription nested polymerase chain reaction targeting the M-segment (NPCR-M) of severe fever with thrombocytopenia syndrome (SFTS) virus. NPCR-M was performed in parallel with three other referenced PCR assays QPCR-S, PCR-M, and NPCR-S to assess their clinical usefulness as routine diagnostic techniques for SFTS. In this multi-centered prospective study, 122 blood samples from 38 laboratory-confirmed SFTS patients and 85 control samples were used. The results demonstrated that QPCR-S and NPCR-S had better sensitivity rate up to 21 days after symptom onset however, the PCR-M showed poor sensitivity after 7 days of symptom onset. Our designed NPCR-M had a higher detection rate up to 40 days from symptom onset and revealed the persistence of SFTSV RNA in the early convalescent phase. No false-positive results were seen for the control samples. Additionally, NPCR-M showed positive results for a sample that initially showed negative results from other PCRs and for many other samples collected in the convalescent phase of SFTS. Our designed nested PCR is suitable for SFTSV detection in patients' blood collected in the acute and early convalescent phase of SFTS, and shows better sensitivity and high specificity even up to 40 days after symptom onset.


Canine hepatobiliary anatomy, physiology and congenital disorders.
The biliary system is an integral component of normal physiologic homeostasis and essential for survival. It acts as a conduit for the removal of detoxified and catabolised compounds as well as aids in fat digestion and absorption. Derangements in this system can have dangerous sequela that are associated with varying degrees of morbidity and mortality. Moreover, abnormalities in development of the biliary system can have varied and unpredictable changes on function and long-term outcome. The aims of this article were to review canine hepatobiliary anatomy, physiology and cholestasis as well as summarise congenital biliary disorders including human corollaries.


Activated alleles of yeast SLN1 increase Mcm1-dependent reporter gene expression and diminish signaling through the Hog1 osmosensing pathway.
Two-component signal transduction systems involving histidine autophosphorylation and phosphotransfer to an aspartate residue on a receiver molecule have only recently been discovered in eukaryotes, although they are well studied in prokaryotes. The Sln1 protein of Saccharomyces cerevisiae is a two-component regulator involved in osmotolerance. Phosphorylation of Sln1p leads to inhibition of the Hog1 mitogen-activated protein kinase osmosensing pathway. We have discovered a second function of Sln1p by identifying recessive activated alleles (designated nrp2) that regulate the essential transcription factor Mcm1. nrp2 alleles cause a 5-fold increase in the activity of an Mcm1-dependent reporter, whereas deletion of SLN1 causes a 10-fold decrease in reporter activity and a corresponding decrease in expression of Mcm1-dependent genes. In addition to activating Mcm1p, nrp2 mutants exhibit reduced phosphorylation of Hog1p and increased osmosensitivity suggesting that nrp2 mutations shift the Sln1p equilibrium toward the phosphorylated state. Two nrp2 mutations map to conserved residues in the receiver domain (P1148S and P1196L) and correspond to residues implicated in bacterial receivers to control receiver phosphorylation state. Thus, it appears that increased Sln1p phosphorylation both stimulates Mcm1p activity and diminishes signaling through the Hog1 osmosensing pathway.


Cytomegalovirus-specific lymphocyte transformations in subjects of different ages with primary immunodeficiency.
The role of specific cell-mediated immune responses in human cytomegalovirus (CMV) infections was studied by an in vitro lymphocyte transformation test, using a whole blood culture. CMV-induced in vitro lymphocyte proliferation was mainly dependent on the presence of sensitized T-cells. Three of five seronegative patients with B-cell deficiency showed positive lymphocyte responses to CMV antigen. In contrast, a patient with ataxia-telangiectasia who was shedding the virus in urine did not show a cellular response to either CMV or phytohemagglutinin. Age-related differences were found in the CMV lymphocyte transformation test. The responses were generally low (P less than 0.025) in infants and higher in older children and adults. CMV lymphocyte transformation responses were lower in 15 children who were excreting CMV in urine when tested than in 17 seropositive children who were not excreting the virus.


Dynamic evaluation of prolactin secretion by successive TRH and metoclopramide stimulations.
We evaluated the clinical applicability of a 2-h test in which the prolactin (Prl) responses to thyrotrophin releasing hormone (TRH)(200 micrograms iv) and the dopamine antagonist, metoclopramide (MC; 10 mg iv) were studied successively, 1 h apart. Nine healthy women were studied with TRH-MC test and with another test in which MC alone was used or the drugs were given in opposite order. The preceding TRH injection did not affect the Prl responses to MC, nor did preceding MC affect the Prl response to TRH. With the TRH-MC test, Prl responses to TRH and MC were significantly lowered in amenorrhoea (N = 8) and hyperprolactinaemia (N = 15) regardless of whether or not treated with bromocriptine, and to MC only in oligomenorrhoea (n = 11). In normoprolactinaemic galactorrhoea (N = 7) the responses were similar as in healthy women. The ratio between Prl responses to TRH and MC was significantly higher in women with bromocriptine-treated hyperprolactinaemia (1.06 +/- 0.8, SD) than in healthy controls (0.34 +/- 0.13). The capacity of pituitary lactotrophs to secrete Prl in response TRH and MC can be evaluated with a 2-h test which has potential for investigation of pituitary Prl dynamics in gynaecological endocrine disorders, particularly in amenorrhoea and hyperprolactinaemia.


Stachyflin and acetylstachyflin, novel anti-influenza A virus substances, produced by Stachybotrys sp. RF-7260. II. Synthesis and preliminary structure-activity relationships of stachyflin derivatives.
Stachyflin and acetylstachyflin, produced by Stachybotrys sp. RF-7260, were found to have potent anti-influenza A virus activity. Stachyflin is a new class of hemagglutinin fusion inhibitors of influenza A virus. Several derivatives were synthesized from acetylstachyflin and subjected to preliminary examination of their structure-activity relationships. Among them, the 3-oxo and 3,8'-dioxo derivatives showed potent antiviral activity similar to stachyflin. The 3-epi derivative was four times less active than stachyflin. Modification of the 6'-hydroxy group and the C-5' position markedly diminished the antiviral activity.


Receptor-mediated sorting of soluble vacuolar proteins: myths, facts, and a new model.
To prevent their being released to the cell exterior, acid hydrolases are recognized by receptors at some point in the secretory pathway and diverted towards the lytic compartment of the cell (lysosome or vacuole). In animal cells, the receptor is called the mannosyl 6-phosphate receptor (MPR) and it binds hydrolase ligands in the trans-Golgi network (TGN). These ligands are then sequestered into clathrin-coated vesicles (CCVs) because of motifs in the cytosolic tail of the MPR which interact first with monomeric adaptors (Golgi-localized, Gamma-ear-containing, ARF-binding proteins, GGAs) and then with tetrameric (adaptin) adaptor complexes. The CCVs then fuse with an early endosome, whose more acidic lumen causes the ligands to dissociate. The MPRs are then recycled back to the TGN via retromer-coated carriers. Plants have vacuolar sorting receptors (VSRs) which were originally identified in CCVs isolated from pea (Pisum sativum L.) cotyledons. It was therefore assumed that VSRs would have an analogous function in plants to MPRs in animals. Although this dogma has enjoyed wide support over the last 20 years there are many inconsistencies. Recently, results have been published which are quite contrary to it. It now emerges that VSRs and their ligands can interact very early in the secretory pathway, and dissociate in the TGN, which, in contrast to its mammalian counterpart, has a pH of 5.5. Multivesicular endosomes in plants lack proton pump complexes and consequently have an almost neutral internal pH, which discounts them as organelles of pH-dependent receptor-ligand dissociation. These data force a critical re-evaluation of the role of CCVs at the TGN, especially considering that vacuolar cargo ligands have never been identified in them. We propose that one population of TGN-derived CCVs participate in retrograde transport of VSRs from the TGN. We also present a new model to explain how secretory and vacuolar cargo proteins are effectively separated after entering the late Golgi/TGN compartments.


Detection of Foreign Antigen-specific CD4(+)Foxp3(+) Regulatory T Cells by MHC Class II Tetramer and Intracellular CD154 Staining.
The unrestricted population of CD4(+)Foxp3(+) regulatory T (Treg) cells, which have been known to control the expression of autoimmune diseases and protective immunity to inflammatory reactions, has led to greater appreciation of functional plasticity. Detecting and/or isolating Ag-specific CD4(+)Foxp3(+) Tregs at the single cell level are required to study their function and plasticity. In this study, we established and compared both MHC class II tetramer and intracellular CD154 staining, in order to detect CD4(+)Foxp3(+) Treg specific for foreign Ag in acute and chronic infections with lymphocytic choriomeningitis virus (LCMV). Our results revealed that MHC class II tetramer staining showed a lower detection rate of LCMV GP66-77-specific CD4(+) T cells because most of MHC class II tetramers were unbound and unstable when combined staining was performed with intracellular cytokines. In contrast, intracellular CD154 staining was revealed to be easier and simple for detecting LCMV GP66-77-specific CD4(+) T cells, compared to MHC class II tetramer staining. Subsequently, we employed intracellular CD154 staining to detect LCMV GP66-77-specific CD4(+)Foxp3(+) Tregs using Foxp3(GFP) knock-in mouse, and found that LCMV GP66-77-specific CD4(+)Foxp3(+) Tregs and polyclonal CD4(+)Foxp3(+) Tregs showed differential expansion in mice infected with LCMV Arms or Cl13 at acute (8 and 13 days pi) and chronic phases (35 days pi). Therefore, our results provide insight into the valuable use of intracellular CD154 staining to detect and characterize foreign Ag-specific CD4(+)Foxp3(+) Treg in various models.


Osseointegration of dental implants in patients undergoing bisphosphonate treatment: a literature review.
Bisphosphonates (BPs) are an important group of drugs used for the treatment of metabolic and oncologic pathologies involving the skeletal system. Osteonecrosis of the jaw (ONJ) is a complication observed in patients using oral or intravenous (IV) BPs. It was suggested that all patients undergoing BP therapy who are expected to receive dental implants should be informed of the possible risks of development of ONJ. The aim of this literature review is to assess the osseointegration of dental implants in patients undergoing BP therapy. The MEDLINE-PubMed databases of The National Library of Medicine, National Institutes of Health, Bethesda, Maryland, were searched for articles addressing the focused question: Can dental implants osseointegrate and remain functionally stable in patients undergoing oral and IV BP therapy? Databases were searched from 1995 up to and including February 2010 using the following terms in different combinations: bisphosphonate, dental implant, immediate-loading, implant survival rate, intravenous, oral, osseointegration, and osteonecrosis. The initial search yielded 89 articles. Scrutiny of the titles and abstracts reduced the number of articles to 12 (seven case reports and five retrospective studies). In 10 studies, the patients were using oral BPs, and in two studies, patients were using IV BPs. Six case reports showed that the placement of implants in patients using BPs could yield a successful osseointegration and function. Four retrospective studies demonstrated that BPs did not have a negative influence on implant success. Two studies showed a negative impact of BPs on implant success. Dental implants can osseointegrate and remain functionally stable in patients using BPs.


Stereo-acuity in patients implanted with multifocal intraocular lenses: is the choice of stereotest relevant?
A randomized and double-blinded study design was implemented to assess the stereo-acuity in patients symmetrically implanted with four types of multifocal intraocular lenses (MIOLs), compared to a monofocal lens (control group). In addition, the influence of the type of test employed for the evaluation of stereo-acuity was explored. Six months after cataract intervention, stereo-acuity was measured with the Titmus and TNO stereotests in 143 patients implanted with one of the following MIOL lens types: hybrid spherical SN60D3, hybrid aspheric SN6AD1, diffractive aspheric ZMA00 and refractive spherical NXG1. A control group implanted with the monofocal aspheric ZA9003 (in which stereo-acuity was measured with a near addition) was also included in the study. Statistically significant better stereo-acuity was found in the monofocal group with both stereotests (except for the SN60D3 group with the Titmus test) (all p < 0.001). No significant differences in stereo-acuity between MIOLs were found using the Titmus test. However, with the TNO, patients implanted with hybrid diffractive MIOLs exhibited statistically significant worse stereo-acuity than those with the refractive design (SN60D3, p < 0.001; SN6AD1, p = 0.006). Patients implanted with MIOLs have worse stereo-acuity than those implanted with monofocal IOLs due to the decrease in retinal image contrast originating in the simultaneous presence of two images. A wavelength-based stereotest such as the TNO induces large differences in image contrast between fellow eyes implanted with diffractive-based MIOLs, which may result in an underestimation of the real stereo-acuity of the patient.


Inadequate values from an interferon-gamma release assay for smear-negative tuberculosis in a high-burden setting.
To examine the usefulness of an interferon-gamma release assay (IGRA) for the diagnosis of smear-negative tuberculosis (TB) in China. A total of 624 patients with presumed pulmonary TB were enrolled prospectively and categorised as smear-negative TB, smear-positive TB or no TB. All patients were tested using T-SPOT.TB. Both the smear-negative and smear-positive TB groups had significantly more spot-forming cells (SFCs) than the no TB group (all P < 0.001), while the smear-negative group had fewer SFCs than the smear-positive TB group (P < 0.001). The specificity of T-SPOT.TB was 60.4% (95%CI 53.4-67.1). The sensitivities of T-SPOT.TB in the smear-negative and smear-positive TB groups were respectively 81.4% (95%CI 75.7-86.0) and 93.2% (95%CI 87.6-96.4). The sensitivity in the smear-negative TB group was much lower than that in the smear-positive TB (P < 0.05). The sensitivity of T-SPOT.TB was lower due the paucibacillary nature of the samples, and the specificity was lower due to the high prevalence of latent tuberculous infection in the smear-negative TB patients. The T-SPOT.TB test should only be used as a supplementary test and not as a single test to rule in or rule out smear-negative TB.


I Am So Tired… How Fatigue May Exacerbate Stress Reactions to Psychological Contract Breach.
Previous research showed that perceptions of psychological contract (PC) breach have undesirable individual and organizational consequences. Surprisingly, the PC literature has paid little to no attention to the relationship between PC breach perceptions and stress. A better understanding of how PC breach may elicit stress seems crucial, given that stress plays a key role in employees' physical and mental well-being. Based on Conservation of Resources Theory, we suggest that PC breach perceptions represent a perceived loss of valued resources, subsequently leading employees to experience higher stress levels resulting from emerging negative emotions. Moreover, we suggest that this mediated relationship is moderated by initial levels of fatigue, due to fatigue lowering the personal resources necessary to cope with breach events. To tests our hypotheses, we analyzed the multilevel data we obtained from two experience sampling designs (Study 1: 51 Belgian employees; Study 2: 53 US employees). Note that the unit of analysis is "observations" rather than "respondents," resulting in an effective sample size of 730 (Study 1) and 374 (Study 2) observations. In both studies, we found evidence for the mediating role of negative emotions in the PC breach-stress relationship. In the second study, we also found evidence for the moderating role of fatigue in the mediated PC breach-stress relationship. Implications for research and practice are discussed.


The Diurnal Profile of Human Basal Pain Sensitivity and Skin Sympathetic Nerve Activity: A Healthy Volunteer Study.
The diurnal rhythm profile of human basal pain sensitivity and its association with sympathetic nerve activity are not fully understood. This study aimed to examine rhythmic changes in experimental pain sensitivity and skin sympathetic nerve activity in healthy volunteers. Thirty healthy volunteers were included in the study. Experimental pain sensitivity, including pressure pain threshold and tolerance, cold pain threshold (CPT) and tolerance, skin sympathetic nerve activity, and cardiovascular parameters (including heart rate, cardiac output, and peripheral vascular resistance) at six time points throughout the day (08:00, 12:00, 16:00, 20:00, 00:00, and 04:00) were sequentially measured. Circadian rhythm analysis was performed on the mean values of the different measurements and individual subjects. Significant differences were found in experimental pain sensitivity, skin sympathetic nerve activity, and non-invasive cardiovascular parameters at different time points (P < 0.05). The minimum measured values of all four types of experimental pain sensitivity were consistently observed at 04:00. Rhythmical analysis showed that the mean values of pressure pain threshold (meta2d P = 0.016) and skin sympathetic nerve activity (meta2d P = 0.039) were significant. Significant diurnal rhythms in pain sensitivity and skin sympathetic nerve activity existed in some individuals but not in others. No significant correlation between experimental pain sensitivity and skin sympathetic nerve activity was found at any time point (P > 0.05). Significant diurnal fluctuations were observed in different pain sensitivities and skin sympathetic nerve activity. No significant correlation between experimental pain sensitivity and sympathetic excitability at different times was found; the reasons for these phenomena remain to be further studied. [www.ClinicalTrials.gov], identifier [ChiCTR2000039709].


[Vaccine against bilharziasis. Strategies and perspectives].
Due to the determination of immunity acquisition mechanisms, a number of studies on experimental models and human populations has made it possible to devise a strategy of vaccination against schistosomiasis, a major worldwide parasitic disease affecting 200 million people. A schistosomial protein, identified as glutathione-S-transferase (SM28GST) has been cloned and expressed in various vectors. Immunization with the recumbent protein results in a significant reduction of the parasitic load, while the fertility of schistosoma and the viability of its eggs are markedly inhibited. Recent experiments performed in primates and bovines have shown that a more than 90% inhibition of the schistosoma's transmission potential can be induced by this vaccine. The important role played by IgA antibodies in the inhibition of the parasite's fertility has led to the development of an oral immunization strategy. In view of the results obtained, the availability, in the near future, of a vaccine protecting human beings against this scourge can be considered realistic.


Effects of stress on the developing brain.
In a complementary article, Judge Cindy Lederman explains the importance of using science to inform family court decisions. Here, Dr. Bruce S. McEwen looks at that science in depth, discussing how early-life stress can lead to long-lasting behavioral, mental, and physical consequences. Fortunately, preventive measures can improve health outcomes, and while interventions for those who have already experienced debilitating early-life stress require considerable effort, they remain possible, thanks to the brain's plasticity.


Prospectives for cell and organ culture systems in the study of pancreatic carcinoma.
Adenocarcinoma of the pancreas is presumed to arise from ductal cell precursors. This brief article questions the definitions on which this assumption is based and suggests possible ways to answer the question since for future studies on the etiology of this cancer it is imperative to study the appropriate cell of origin. Possible approaches for future work on pancreatic carcinogenesis utilizing cell and organ culture systems are discussed.


Shortening-induced residual force depression in humans.
When an isometric muscle contraction is immediately preceded by an active shortening contraction, a reduction in steady-state isometric force is observed relative to an isometric reference contraction at the same muscle length and level of activation. This shortening-induced reduction in isometric force, termed "residual force depression" (rFD), has been under investigation for over a half century. Various experimental models have revealed the positive relationship between rFD and the force and displacement performed during shortening, with rFD values ranging from 5 to 39% across various muscle groups, which appears to be due to a stress-induced inhibition of cross-bridge attachments. The current review will discuss the findings of rFD in humans during maximal and submaximal contractions.


Diagnostic accuracy of the repeatable battery of the assessment of neuropsychological status update, Spanish version, in predicting Alzheimer's disease among Hispanic older adults in the United States reporting memory problems.
There are few standardized, Spanish-language diagnostic tools to help identify Hispanic persons at early stages of Alzheimer's disease (AD). This study evaluated the accuracy of the Spanish version of the Repeatable Battery for the Assessment of Neuropsychological Status-Update (RBANS) in predicting AD in older Hispanic adults in the United States reporting memory problems. We analyzed data from age, sex, and education level propensity score-matched Hispanic memory clinic patients with (n = 38) and without (n = 38) a clinical diagnosis of AD. Estimates of diagnostic accuracy included sensitivity, specificity, predictive value, and receiver operating characteristic analysis. After controlling for sex and matched pairs, the Total Scale score [area under curve (AUC) = 0.7417] and the Immediate (AUC = 0.7258) and Delayed (AUC = 0.7735) Memory index scores provided better estimates of diagnostic accuracy than Language, Attention, and Visuospatial/Constructional index scores. A minus 2-standard deviation (SD) cut point enhanced the predictive probability of the Delayed Memory index score. A cut point of -1.5 SD optimized the predictive probability of the Total Scale score. These results suggest that optimal cutoff values for the RBANS Delayed Memory index and Total Scale scores that may help identify Hispanic patients with AD as part of a comprehensive diagnostic AD assessment.


Electronegative low density lipoprotein induces renal apoptosis and fibrosis: STRA6 signaling involved.
Dyslipidemia has been proven to capably develop and aggravate chronic kidney disease. We also report that electronegative LDL (L5) is the most atherogenic LDL. On the other hand, retinoic acid (RA) and RA receptor (RAR) agonist are reported to be beneficial in some kidney diseases. "Stimulated by retinoic acid 6" (STRA6), one retinol-binding protein 4 receptor, was recently identified to regulate retinoid homeostasis. Here, we observed that L5 suppressed STRA6 cascades [STRA6, cellular retinol-binding protein 1 (CRBP1), RARs, retinoid X receptor α, and retinol, RA], but L5 simultaneously induced apoptosis and fibrosis (TGFβ1, Smad2, collagen 1, hydroxyproline, and trichrome) in kidneys of L5-injected mice and L5-treated renal tubular cells. These L5-induced changes of STRA6 cascades, renal apoptosis, and fibrosis were reversed in kidneys of LOX1(-/-) mice. LOX1 RNA silencing and inhibitor of c-Jun N-terminal kinase and p38MAPK rescued the suppression of STRA6 cascades and apoptosis and fibrosis in L5-treated renal tubular cells. Furthermore, crbp1 gene transfection reversed downregulation of STRA6 cascades, apoptosis, and fibrosis in L5-treated renal tubular cells. For mimicking STRA6 deficiency, efficient silencing of STRA6 RNA was performed and was found to repress STRA6 cascades and caused apoptosis and fibrosis in L1-treated renal tubular cells. In summary, this study reveals that electronegative L5 can cause kidney apoptosis and fibrosis via the suppression of STRA6 cascades, and implicates that STRA6 signaling may be involved in dyslipidemia-mediated kidney disease.


Theodor Escherich: the first pediatric infectious diseases physician?
The career of Theodor Escherich (1857-1911) qualifies him as the first pediatric infectious diseases physician. His landmark bacteriologic studies identified the common colon bacillus (now known as Escherichia coli), and he was very committed to pediatrics, serving as chairman of several prominent departments of pediatrics, including the Department of Pediatrics at the University of Vienna and St. Anna's Children's Hospital in Vienna, Austria, arguably Europe's most prestigious pediatric position.


Lung cGMP release subsequent to NO inhalation in pulmonary hypertension: responders versus nonresponders.
Inhalation of nitric oxide (NO) is widely employed for the assessment of pulmonary vasoresponsiveness in pulmonary hypertension (PH). However, the reasons for the huge differences in vascular reactivity to NO between patients are unknown, and the role of NO-induced cyclic guanosine monophosphate (cGMP) is unclear. Twenty patients with severe precapillary PH were investigated. Thirty-six Swan-Ganz catheter investigations were performed and the study subjects were tested for responses to NO inhalation. This included an assessment of pulmonary and systemic arterial plasma cGMP and atrial natriuretic peptide (ANP) levels. A significant NO response (pulmonary vascular resistance (PVR) decrease >20%) was noted in nine of 20 patients (45%) during the first catheterization. A highly significant correlation between baseline plasma cGMP and ANP levels with PVR was observed (r=0.62 and r=0.66, respectively; p<0.0001). In response to NO, systemic and mixed venous cGMP levels increased from 13.9 +/- 1.28 nM and 12.75 +/- 0.99 nM to 79.23 +/- 4.99 nM and 55.25 +/- 4.41 nM (p<0.001), respectively, accompanied by the appearance of a marked transpulmonary cGMP gradient. Although in the responder group ANP levels were significantly reduced after NO inhalation, no significant correlation was observed to the extent of PVR reduction. The magnitude of the NO-elicited cGMP response did not discriminate between haemodynamic responders and nonresponders. This study concludes that plasma cyclic guanosine monophosphate levels are significantly correlated with the severity of disease in pulmonary arterial hypertension. Nitric oxide inhalation provokes a prompt increase in cyclic guanosine monophosphate secretion, but the magnitude of this release is not linked with a decrease in pulmonary vascular resistance.


AAV-CRISPR Gene Editing Is Negated by Pre-existing Immunity to Cas9.
Adeno-associated viral (AAV) vectors are a leading candidate for the delivery of CRISPR-Cas9 for therapeutic genome editing in vivo. However, AAV-based delivery involves persistent expression of the Cas9 nuclease, a bacterial protein. Recent studies indicate a high prevalence of neutralizing antibodies and T cells specific to the commonly used Cas9 orthologs from Streptococcus pyogenes (SpCas9) and Staphylococcus aureus (SaCas9) in humans. We tested in a mouse model whether pre-existing immunity to SaCas9 would pose a barrier to liver genome editing with AAV packaging CRISPR-Cas9. Although efficient genome editing occurred in mouse liver with pre-existing SaCas9 immunity, this was accompanied by an increased proportion of CD8+ T cells in the liver. This cytotoxic T cell response was characterized by hepatocyte apoptosis, loss of recombinant AAV genomes, and complete elimination of genome-edited cells, and was followed by compensatory liver regeneration. Our results raise important efficacy and safety concerns for CRISPR-Cas9-based in vivo genome editing in the liver.


Hypocalcaemia following laryngectomy: prevalence and risk factors.
To establish the prevalence of hypocalcaemia following laryngectomy and demonstrate that total thyroidectomy is a risk factor. A retrospective cohort study was conducted that included all patients who underwent total laryngectomy from 1st January 2006 to 1st August 2017. Exclusion criteria were: pre-operative calcium derangement, previous thyroid or parathyroid surgery, concurrent glossectomy, pharyngectomy, or oesophagectomy. Ninety patients were included. Sixteen patients had early hypocalcaemia (18 per cent), seven had protracted hypocalcaemia (8 per cent) and six had permanent hypocalcaemia (10 per cent). Exact logistic regression values for hypocalcaemia following total thyroidectomy compared to other patients were: early hypocalcaemia, odds ratio = 15.5 (95 per cent confidence interval = 2.2-181.9; model p = 0.002); protracted hypocalcaemia, odds ratio = 13.3 (95 per cent confidence interval = 1.5-117.1; model p = 0.01); and permanent hypocalcaemia, odds ratio = 22.7 (95 per cent confidence interval = 1.9-376.5; model p = 0.005). This is the largest study to investigate the prevalence of hypocalcaemia following laryngectomy and the first to include follow up of longer than three months. Total thyroidectomy significantly increased the risk of hypocalcaemia at all time frames and independent of other variables.


A high anisotropy barrier in a sulfur-bridged organodysprosium single-molecule magnet.
The sulfur-bridged dimers [{Cp'(2)Ln(μ-SSiPh(3))}(2)] (Ln=Gd (1), Dy (2); Cp'=η(5)-C(5)H(4)Me) were synthesized by the transmetalation reactions between [Cp'(3)Ln] and Ph(3)SiSLi. Compound 2 is a single-molecule magnet with slow relaxation of magnetization up to 40 K and an anisotropy barrier of U(eff) =133 cm(-1). Insight into the SMM properties of 2 and closely related SMMs has been obtained using ab initio calculations.


IRF4 and IRGs Delineate Clinically Relevant Gene Expression Signatures in Systemic Lupus Erythematosus and Rheumatoid Arthritis.
Introduction: Overactivation of the type I interferon (IFN) signature has been observed in several systemic autoimmune conditions, such as Systemic Lupus Erythematosus (SLE) or Rheumatoid Arthritis (RA). Impaired control of Interferon-Responding Genes (IRGs) expression by their regulatory mechanisms, including Interferon Regulatory Factors (IRFs), may underlie these findings and it may explain the heterogeneity observed among these conditions. In the present study we aimed to evaluate the associations between IRF4 gene expression and those of IRGs in SLE and RA patients to gain insight about its links with the IFN signature as well as to explore the potential clinical relevance of these associations. Methods: The gene expression of IRF4 and IRGs (IFI44, IFI44L, IFI6, and MX1) in peripheral blood was analyzed in 75 SLE patients, 98 RA patients, and 28 healthy controls. A group of 13 biological-naïve RA patients was prospectively followed upon TNFα-blockade. The associations among IRF4 and IRGs were evaluated by principal component analyses (PCA), correlations and network analyses. Publicly available datasets were used for replication. Results: A broad activation of IRGs was observed in autoimmune patients, although certain heterogeneity can be distinguished, whereas IRF4 was only upregulated in RA. The differential expression of IRF4 in RA was then confirmed in publicly available gene expression datasets. PCA revealed different associations among IRF4 and IRGs in each condition, which was later confirmed by correlation and network analyses. Cluster analysis identified 3 gene expression signatures on the basis of IRF4 and IRGs expression which were differentially used by SLE and RA patients. Cluster III was associated with markers of disease severity in SLE patients. Cluster II, hallmarked by IRF4 upregulation, was linked to clinical stage and mild disease course in RA. TNFα-blockade led to changes in the association between IRF4 and IRGs, whereas increasing IRF4 expression was associated with a good clinical outcome in RA. Conclusions: The differential expression of IRF4 and IRGs observed in SLE and RA can delineate gene expression signatures associated with clinical features and treatment outcomes. These results support a clinically-relevant phenomenon of shaping of the IFN signature by IRF4 in autoimmune patients.


Submicron-precision particle characterization in microfluidic impedance cytometry with double differential electrodes.
Submicron-precision particle characterization is crucial for counting, sizing and identifying a variety of biological particles, such as bacteria and apoptotic bodies. Microfluidic impedance cytometry has been attractive in current research for microparticle characterization due to its advantages of label-free detection, ease of miniaturization and affordability. However, conventional electrode configurations of three electrodes and floating electrodes have not yet demonstrated the capability of probing submicron particles or microparticles with a submicron size difference. In this study, we present a label-free high-throughput (∼800 particles per second) impedance-based microfluidic flow cytometry system integrated with a novel design of a double differential electrode configuration, enabling submicron particle detection (down to 0.4 μm) with a minimum size resolution of 200 nm. The signal-to-noise ratio has been boosted from 13.98 dB to 32.64 dB compared to a typical three-electrode configuration. With the proposed microfluidic impedance cytometry, we have shown results of sizing microparticles that accurately correlate with manufacturers' datasheets (R2 = 0.99938). It also shows that population ratios of differently sized beads in mixture samples are consistent with the results given by commercial fluorescence-based flow cytometry (within ∼1% difference). This work provides a label-free approach with submicron precision for sizing and counting microscale and submicron particles, and a new avenue of designing electrode configurations with a feature of suppressing the electrical noise for accomplishing a high signal-to-noise ratio in a wide range of frequencies. This novel double differential impedance sensing system paves a new pathway for real-time analysis and accurate particle screening in pathological and pharmacological research.


Application of Machine Learning Algorithms for Tool Condition Monitoring in Milling Chipboard Process.
In this article, we present a novel approach to tool condition monitoring in the chipboard milling process using machine learning algorithms. The presented study aims to address the challenges of detecting tool wear and predicting tool failure in real time, which can significantly improve the efficiency and productivity of the manufacturing process. A combination of feature engineering and machine learning techniques was applied in order to analyze 11 signals generated during the milling process. The presented approach achieved high accuracy in detecting tool wear and predicting tool failure, outperforming traditional methods. The final findings demonstrate the potential of machine learning algorithms in improving tool condition monitoring in the manufacturing industry. This study contributes to the growing body of research on the application of artificial intelligence in industrial processes. In conclusion, the presented research highlights the importance of adopting innovative approaches to address the challenges of tool condition monitoring in the manufacturing industry. The final results provide valuable insights for practitioners and researchers in the field of industrial automation and machine learning.


Scaling dependence and tailoring of the pinning field in FePt-based exchange coupled composite media.
We studied exchange coupled composite (ECC) media with an out-of-plane easy axis consisting of hard magnetic L1(0) chemically ordered FePtCu alloy films and magnetically softer [Co/Pt](N) multilayer stacks. By tailoring the structural properties of the ternary FePtCu alloy and [Co/Pt](N) multilayers, we can tune the magnetic parameters of the composite in a wide range. This allowed us to address experimentally one of the most crucial properties determining the performance of ECC media, namely the pinning field of the magnetic domain wall present at the interface between the hard and soft layers. We demonstrate that the pinning field is proportional to the difference of the magnetic anisotropy constants of the hard and soft layers, which confirms the theoretical predictions. Furthermore, we show that the pinning field can be efficiently decreased after an additional annealing step. Transmission electron microscopy investigations indicated that the origin of the observed effect is due to a heat-induced phase transformation of iron oxide present at the interface between the hard and soft layers. This study reveals that tailoring the properties of the hard/soft interface is another efficient tuning knob for optimization of the performance of ECC media.


New covalent capture probes for imaging and therapy, based on a combination of binding affinity and disulfide bond formation.
We describe the synthesis and development of new reactive DOTA-metal complexes for covalently targeting engineered receptors in vivo, which have superior tumor uptake and clearance properties for biomedical applications. These probes are found to clear efficiently through the kidneys and minimally through other routes, but bind persistently in the tumor target. We also explore the new technique of Cerenkov luminescence imaging to optically monitor radiolabeled probe distribution and kinetics in vivo. Cerenkov luminescence imaging uniquely enables sensitive noninvasive in vivo imaging of a β(-) emitter such as (90)Y with an optical imager.


Tetrazine-based cycloadditions: application to pretargeted live cell imaging.
Bioorthogonal tetrazine cycloadditions have been applied to live cell labeling. Tetrazines react irreversibly with the strained dienophile norbornene forming dihydropyrazine products and dinitrogen. The reaction is high yielding, selective, and fast in aqueous media. Her2/neu receptors on live human breast cancer cells were targeted with a monoclonal antibody modified with a norbornene. Tetrazines conjugated to a near-infrared fluorochrome selectively and rapidly label the pretargeted antibody in the presence of serum. These findings indicate that this chemistry is suitable for in vitro labeling experiments, and suggests that it may prove a useful strategy for in vivo pretargeted imaging under numerous modalities.


Double-crosslinked PNIPAM-based hydrogel dressings with adjustable adhesion and contractility.
Rapid post-wound closure is necessary to avoid wound infection and promote scar-free healing when skin trauma occurs. In this study, new types of hydrogel dressings with adjustable contractility were fabricated based on N-isopropyl acrylamide/sodium alginate/graphene oxide (P/SA/GO). Then, the chitosan (CS) solution was used as a bridging polymer to achieve tissue adhesion to the hydrogel. The results show that the hydrogel based on poly(N-isopropyl acrylamide) (PNIPAM) not only has the ability to self-shrink but also can adjust the rate of shrinkage through near-infrared thermal stimulation. At the same time, high adhesion strength (7.86 ± 1.22 kPa) between the tissue and the dressing is achieved through the introduction of bridging polymers (CS), and the coating area of the bridging polymer can be adjusted to achieve regional adhesion. The mouse total skin defects experiments have shown that sutures-free wound closure in the early stages of wound healing could be obtained by adjusting the material temperature. Besides, the dressings can promote scar-free wound healing by reducing inflammatory cell infiltration and collagen deposition. These results indicate that double-crosslinked PNIPAM-based hydrogel dressings with adjustable adhesion and contractility proposed in this study provide a candidate material for achieving trackless wound healing.


[An electron microscopic study of the structure of the rat liver under the action of antimembrane antibodies].
Introduction of heteroantibodies (4 mg/100 g) to plasma membranes of hepatocytes into portal vein of the rat liver caused (already within 5 min) changes in the liver ultrastructure with their intensification by the 30th min. Specificity of the action of antimembrane antibodies (Am) was revealed as against the action of antibodies to summary antigens (As) of the liver. It is shown that Am change, mainly, sinusoidal and lateral regions of the hepatocyte membrane, not disturbing essentially structure of its cytoplasma, intracellular organelles and biliary canaliculi, endotheliocytes and stellate reticuloendotheliocytes, that is observed while introducing As to the liver. Membrane tracer--colloidal lanthanum has revealed that Am increases permeability of hepatocyte plasmalemma to a less degree than As of the liver.


[Effect of combined irradiation and hyperthermia on the creatine kinase activity of rat tissues].
Effects of heating (36 degrees, relative air humidity 80-90%, 4 hours) and joint beta (85Kr)-X-ray-irradiation of rats, given separately and simultaneously, in doses 2.5 krad+ + 25 r, 2.5 krad + 50 r, 3.05 krad + 100 r, 4.45 krad + 250 r, 7.4 krad + 400 r on creatine kinase activity in brain and liver were investigated 5, 12, 19 and 26 days after the exposure. It was discovered that the state of thermal stress reduced radiosensitiveness of enzymatic activity in brain and burdened the consequences of irradiation when tested by the liver activity.


Chemicals of predatory mosquitofish (Gambusia affinis) influence selection of oviposition site by Culex mosquitoes.
Ovipositing insects may avoid aquatic sites where there is high predation risk to their offspring, but the proximate mechanisms that mediate avoidance behavior are poorly resolved. We conducted an experiment to determine whether mosquitoes would reduce oviposition rates in pools containing chemicals of the mosquitofish (Gambusia affinis), a voracious predator that is widely employed to control mosquitoes. Experimental treatments consisted of outdoor pools that contained known concentrations of fish chemicals (low, medium, or high) or no fish chemicals (control). The pools were arranged in a randomized block design, and the number of mosquito larvae in each pool served as the response variable to estimate relative oviposition rate. Members of the Culex pipiens complex were the main colonizers of the pools. The mean number of larvae per pool differed among treatments (P = 0.026) and was about three times greater in control pools compared with those receiving medium and high concentrations of fish chemicals. Pairwise comparisons indicate that only medium and high treatments differed significantly from controls, suggesting that a threshold concentration exists below which mosquitoes cannot reliably detect predators. Our data suggest that the effectiveness of Gambusia affinis in controlling mosquitoes may be compromised if adult mosquitoes respond to fish stocking by shifting to nearby breeding sites that lack fish. We discuss issues conceming the use of Gambusia in biological control programs within the context of these new findings.


Activation of the embryonic genome.
Activation of its genome is amongst the essential task the embryo has to undertake following fertilization of the egg. In animal and plants, this activation follows a period of transcriptional silence, which is made necessary by the requirement for an almost complete and functional reprogramming of the DNA coming from both gametes. The process by which DNA is silenced, reprogrammed and reactivated is not fully understood yet but progresses are being made, especially with the help of genomic tools. This review will focus on the recent discoveries made in different animal models and more specifically on the efforts made to further characterize the event of maternal to embryonic transition in bovine embryos.


Locally accessible information: how much can the parties gain by cooperating?
We investigate measurements of bipartite ensembles restricted to local operations and classical communication and find a universal Holevo-like upper bound on the locally accessible information. We analyze our bound and exhibit a class of states which saturate it. Finally, we link the bound to the problem of quantification of the nonlocality of the operations necessary to extract locally inaccessible information.


Depletion of tumor-associated macrophages slows the growth of chemically induced mouse lung adenocarcinomas.
Chronic inflammation is a risk factor for lung cancer, and low-dose aspirin intake reduces lung cancer risk. However, the roles that specific inflammatory cells and their products play in lung carcinogenesis have yet to be fully elucidated. In mice, alveolar macrophage numbers increase as lung tumors progress, and pulmonary macrophage programing changes within 2 weeks of carcinogen exposure. To examine how macrophages specifically affect lung tumor progression, they were depleted in mice bearing urethane-induced lung tumors using clodronate-encapsulated liposomes. Alveolar macrophage populations decreased to ≤50% of control levels after 4-6 weeks of liposomal clodronate treatment. Tumor burden decreased by 50% compared to vehicle treated mice, and tumor cell proliferation, as measured by Ki67 staining, was also attenuated. Pulmonary fluid levels of insulin-like growth factor-I, CXCL1, IL-6, and CCL2 diminished with clodronate liposome treatment. Tumor-associated macrophages expressed markers of both M1 and M2 programing in vehicle and clodronate liposome-treated mice. Mice lacking CCR2 (the receptor for macrophage chemotactic factor CCL2) had comparable numbers of alveolar macrophages and showed no difference in tumor growth rates when compared to similarly treated wild-type mice suggesting that while CCL2 may recruit macrophages to lung tumor microenvironments, redundant pathways can compensate when CCL2/CCR2 signaling is inactivated. Depletion of pulmonary macrophages rather than inhibition of their recruitment may be an advantageous strategy for attenuating lung cancer progression.


Experimental Study on the Physicochemical Properties of Asphalt Modified by Different Anti-Stripping Agents and Their Moisture Susceptibility with Aggregates.
Erosion and the stripping effect of moisture on asphalt mixtures is one of the main reasons for the shortened service life of asphalt pavements. The common mean of preventing asphalt pavements from being damaged by moisture is adding anti-stripping agents (ASAs) to asphalt mixtures. However, the effect regularity and mechanism of anti-stripping agents on the physicochemical properties of asphalt is not exactly defined. This study compared the physical properties of ASA-modified asphalt (AMAs) to determine the optimal dosage and investigated the rheological and adhesion properties. Based on the roller bottle method and water immersion method, the moisture susceptibility of AMAs with three particle sizes was investigated. The results showed that the modification of asphalt using anti-stripping agents was a physical modification. At the optimum dosage of anti-stripping agents (0.3%), the basic physical properties of AMA1 were the most desirable. ASA2 increased the resistance of asphalt for deformation at high temperature by 46%, and AMA3 had the best low-temperature performance. ASAs enhanced the dispersed and polar components in the asphalt binder, improving the adhesion energy of asphalt. AMA3 had the strongest adhesion to the aggregate, with an increase in adhesion work by 2.8 times and a 45% of increase in ER value. This was attributed to ASA3 containing with a large number of metal cations and polar functional groups. It was shown that ASAs provided the most improvement in the anti-stripping performance of asphalt mixtures with 9.5-13.2 mm particles. The amide ASA, phosphate ASA and aliphatic amine ASA improved the water damage resistance of asphalt by 65%, 45% and 78%, respectively. This study can help engineers realize the effects of different types of ASAs on the physicochemical properties of asphalt and select the most suitable type of ASAs according to the service requirements.