Immunoglobulin levels in panic disorder patients.
The study was conducted to evaluate the serum immunoglobulin levels in patients suffering from panic disorder and to assess the relationship between the changes of immunoglobulin levels and the socioeconomic parameters, as well as nutritional status. 54 panic patients were randomly selected from the Department of Psychiatry, Bangabandhu Sheikh Mujib Medical University (BSMMU) and Dhaka Medical College Hospital, Bangladesh. Fifty two, age and gender matched healthy volunteers (42 males and 10 females, mean age of 30 ± 6 yrs) were also enrolled in this study. Immunoglobulin levels were measured by turbidimetry method using immunoglobulin kits. It was found that the mean serum immunoglobulin concentrations of IgG, IgM and IgA of panic disorder patients were 0.999±0.26 (g/L), 0.1±0.028 (g/L) and 0.194±0.066 (g/L) respectively whereas the values were 1.24± 0.39 ( g/L ), 0.096±0.022 ( g/L) , 0.194±0.053 (g/L) in healthy volunteers. IgG level in panic disorder patient was found significantly (p <0.05) lower than that of the controls but the change in concentration of IgM and IgA were not significant (p=0.497, p=0.962). Socioeconomic data reveals that most of the patients were from lower income group and educated. BMI (Mean±SD) of the patients (22.62 ± 3.74 kg/m2) and controls (23.74 ± 2.71 kg/m2) were well within the normal range. From correlative analysis it has been found that income has significant effect (p=0 .047) on the change of the serum IgG level in panic disorder patient and it was also been justified by the regression analysis (p=0.049). This finding may play a key role in the diagnosis and treatment of the panic disorder patients. Further studies have been suggested with a large number of populations to confirm these findings.


[Drop in suicide rate after first psychosis: a comparison with the situation two decades ago].
Over two decades ago a study reported that a percentage as high as 11% of the patients with a psychotic disorder in the northern Netherlands died by committing suicide. The primary objective of the current study was to replicate these findings in the same geographical area. A secondary objective was to investigate potential predictors of suicide. Retrospective cohort study. From patient files we identified the current status of all patients who experienced their first episode of psychosis and participated in the Psychosis Recent Onset Groningen-Survey (PROGR-S) between 2000 and 2009. The PROGR-S is a diagnostic protocol for persons with suspected first psychosis. Kaplan-Meier survival analysis was used to identify the current suicide risk in this group and this was compared with the 1973-1988 patients using a log rank test. We also determined the standardized mortality rate in relation to the general population. Cox regression analysis was used to assess significant predictors of suicide. The suicide risk dropped drastically over the course of two decades from 11% to 2.4%. Older age was the strongest predictor of suicide. The standardized mortality rate was 41.6. Despite the significant decline in suicide after a first episode of psychosis, the risk of suicide should be given the highest priority in psychiatric psychosis care, as it is very high at 4162% compared with the suicide risk in general population.


Preventing new sensitization and asthma onset by allergen immunotherapy: the current evidence.
Specific allergen immunotherapy is considered a key candidate for a successful preventive intervention in atopic diseases. The strong association of atopic manifestations such as rhinitis and asthma with atopic sensitizations (specific serum IgE) provide a rationale for early intervention in childhood and adolescence. Currently, the documentation of the disease-modifying intervention effects is limited to the secondary prevention of asthma symptoms in children with allergic rhinoconjunctivitis. These effects appear to be rather allergen specific than nonspecific. Documentation on disease modification including a reduction of asthma symptoms in children, particularly with grass pollen tablets has become quite robust. It is not clear up to now, if the new onset of allergic sensitizations can be modified. So far data on primary prevention are not conclusive.


Hyaluronic acid-modified bioadhesive liposomes as local drug depots: effects of cellular and fluid dynamics on liposome retention at target sites.
Bioadhesive liposomes, in which hyaluronic acid is the surface-anchored bioadhesive ligand, are being tested in vitro in order to evaluate their bioadhesivity. The first part of the test, binding to monolayers of cells modeling the in vivo designated sites under static conditions, was reported in a previous communication. This communication reports the results of the second and third parts of the test, which consist of evaluating the retention of bound liposomes in the face of tissue-related events such as cell migration, proliferation, and death, and under fluid flow. The in vivo-designated binding sites for the bioadhesive liposomes were modeled, as before, by monolayers of the A431 cell line served. A setup for perfusing a culture flask containing a monolayer of cells was devised for measuring the retention under fluid flow. The major findings are: (1) Over a selected tested period of 28 h, the cell cultures mimicked the tissue-related events described above, whether they did or did not receive a dose of liposomes. Over the same period and throughout these events, the bioadhesive liposomes remained bound at equilibrium-like levels, in the range of 0.03 ng lipid/10(5) cells. (2) Fluid flowed over a cell monolayer dosed with bioadhesive liposomes swept away part of the dose. This loss was due mostly to that fraction of the dose which was in excess of the binding capacity of the monolayer, and occurred over the first 15 min of flow. Thereafter, the cell monolayer retained the bound liposomes, at equilibrium-like levels and with no further loss, even for the longest flow period tested (45 min of continuous flow) and under a flow rate of 0.64 ml/min. This study, together with previous results, shows that the hyaluronic acid-modified liposomes meet all parts of the bioadhesivity test. We therefore find merit in their further investigation.


[Interventional endoscopy for benign and malignant bile duct strictures].
During the past years several endoscopic and interventional techniques have been developed for the treatment of bile duct strictures and have had a strong impact on therapeutic regimens. Benign stenoses of the bile duct are mainly caused by cholecystectomy or liver resection or by inflammatory diseases. Insertion of an endoprosthesis insertion or balloon dilation is clinically successful in 60 to 90 % of these patients and will result in adequate opening of the stricture. To date, only bile duct stenosis in chronic pancreatitis are not improved satisfactorily by endoscopy. The insertion of an endoprosthesis is a cornerstone in the treatment of malignant obstructive jaundice in patients with cancer. Several comparative studies have demonstrated the advantages of self-expanding metal stents (SEMS) over plastic prostheses in terms of patency. A selective use of SEMS is mandatory, as the costs for SEMS are high and many patients with malignant jaundice will die with their first plastic prosthesis in situ without stent occlusion. In patients with hilar cholangiocarcinoma, the combination of photodynamic therapy and endoprosthesis insertion might result in a survival advantage. The use of bioabsorbable stent materials or coating of the stent with antiproliferative drugs will improve the treatment results in the future.


Effective functional mapping of fMRI data with support-vector machines.
There is a growing interest in using support vector machines (SVMs) to classify and analyze fMRI signals, leading to a wide variety of applications ranging from brain state decoding to functional mapping of spatially and temporally distributed brain activations. Studies so far have generated functional maps using the vector of weight values generated by the SVM classification process, or alternatively by mapping the correlation coefficient between the fMRI signal at each voxel and the brain state determined by the SVM. However, these approaches are limited as they do not incorporate both the information involved in the SVM prediction of a brain state, namely, the BOLD activation at voxels and the degree of involvement of different voxels as indicated by their weight values. An important implication of the above point is that two different datasets of BOLD signals, presumably obtained from two different experiments, can potentially produce two identical hyperplanes irrespective of their differences in data distribution. Yet, the two sets of signal inputs could correspond to different functional maps. With this consideration, we propose a new method called Effect Mapping that is generated as a product of the weight vector and a newly computed vector of mutual information between BOLD activations at each voxel and the SVM output. By applying this method on neuroimaging data of overt motor execution in nine healthy volunteers, we demonstrate higher decoding accuracy indicating the greater efficacy of this method.


Inflammatory pathway genes associated with inter-individual variability in the trajectories of morning and evening fatigue in patients receiving chemotherapy.
Fatigue, a highly prevalent and distressing symptom during chemotherapy (CTX), demonstrates diurnal and interindividual variability in severity. Little is known about the associations between variations in genes involved in inflammatory processes and morning and evening fatigue severity during CTX. The purposes of this study, in a sample of oncology patients (N=543) with breast, gastrointestinal (GI), gynecological (GYN), or lung cancer who received two cycles of CTX, were to determine whether variations in genes involved in inflammatory processes were associated with inter-individual variability in initial levels as well as in the trajectories of morning and evening fatigue. Patients completed the Lee Fatigue Scale to determine morning and evening fatigue severity a total of six times over two cycles of CTX. Using a whole exome array, 309 single nucleotide polymorphisms SNPs among the 64 candidate genes that passed all quality control filters were evaluated using hierarchical linear modeling (HLM). Based on the results of the HLM analyses, the final SNPs were evaluated for their potential impact on protein function using two bioinformational tools. The following inflammatory pathways were represented: chemokines (3 genes); cytokines (12 genes); inflammasome (11 genes); Janus kinase/signal transducers and activators of transcription (JAK/STAT, 10 genes); mitogen-activated protein kinase/jun amino-terminal kinases (MAPK/JNK, 3 genes); nuclear factor-kappa beta (NFkB, 18 genes); and NFkB and MAP/JNK (7 genes). After controlling for self-reported and genomic estimates of race and ethnicity, polymorphisms in six genes from the cytokine (2 genes); inflammasome (2 genes); and NFkB (2 genes) pathways were associated with both morning and evening fatigue. Polymorphisms in six genes from the inflammasome (1 gene); JAK/STAT (1 gene); and NFkB (4 genes) pathways were associated with only morning fatigue. Polymorphisms in three genes from the inflammasome (2 genes) and the NFkB (1 gene) pathways were associated with only evening fatigue. Taken together, these findings add to the growing body of evidence that suggests that morning and evening fatigue are distinct symptoms.


Magnetic field-induced assembly of oriented superlattices from maghemite nanocubes.
Tailoring the structure of nanocrystal superlattices is an important step toward controlled design of novel nanostructured materials and devices. We demonstrate how the long-range order and macroscopic dimensions of magnetic nanoparticle arrays can be controlled by the use of a modulated magnetic field. Inducing a dipolar attraction during the initial stage of the drying-mediated self-assembly process was sufficient to assemble the superparamagnetic oleate-capped maghemite nanocubes into large and defect-free superstructures with both translational and orientational order. The characteristic dimensions of the superlattice are controlled by the particle concentration as well as the duration of the applied magnetic field. The superparamagnetic maghemite nanocubes assemble into large and highly oriented thin arrays by applying the magnetic field perpendicular to the substrate surface only during the initial phase of drying-mediated self-assembly. Micrometer-sized and thick three-dimensional mesocrystals are obtained when the drying dispersion is subjected to an external magnetic field of moderate strength for the entire duration of the assembly process. The discovery of how translational and orientation order of nanocrystal superlattices can be induced by a temporal modulation of an anisotropic interparticle force offers new insight on the importance of the initial nucleation stage in the self-assembly process and suggests new routes for controlled self-assembly of dipolar nanocrystals.


Importance of the Complement Alternative Pathway in Serum Chemotactic Activity During Sepsis.
Serum chemotactic activity is important in regulating neutrophil migration. The ability of neutrophils to migrate to infectious site is crucial for host effective pathogen control, but unregulated neutrophil activation can also cause tissue damage. During bacterial sepsis, the complement alternative pathway (AP) is massively activated in blood and tissues and reportedly contributes to sepsis pathogenesis. Complement factor B (FB) is an essential component of the AP activation. However, the impact of FB/AP activation on blood chemotactic activity during bacterial infection is unclear. In this study, we found that sera of septic mice following cecal ligation and puncture (CLP) had much higher chemotactic activities on neutrophils than those of sham animals. Compared with wild-type (WT) mice, FB mice had significantly attenuated serum chemotactic activity, under both nonseptic and septic conditions. Moreover, sera with the activated AP by zymosan and cobra venom factor (CVF) in vitro induced a significant increase in neutrophil migration compared with sera without the AP activation. Complement activation generates complement cleavage fragment such as Ba, C3a, and C5a. To delineate the contribution of these downstream effectors, we incubated AP-active sera (AP activated by zymosan/CVF) or sera from sham and septic mice with anti-C5a or mAb1379 (anti-Ba) neutralizing antibody. We found that anti-C5a, but not mAb1379, markedly attenuated the neutrophil chemotactic effect of the AP-activated sera and that of septic sera. Taking together, these data suggest that the complement AP activation during bacterial sepsis plays a pivotal role in promoting blood chemotactic activity through a C5a-dependent mechanism.


Structures and diverse functions of frog growth hormone-releasing peptide (fGRP) and its related peptides (fGRP-RPs): a review.
A new Arg-Phe-NH(2) (RFamide) peptide has been discovered in the amphibian hypothalamus. The cell bodies and terminals containing this peptide were localized in the suprachiasmatic nucleus and median eminence, respectively. This peptide was further revealed to have a considerable growth hormone (GH)-releasing activity in vitro and in vivo and hence designated as frog GH-releasing peptide (fGRP). Molecular cloning of cDNA encoding the fGRP precursor polypeptide revealed that it encodes fGRP and its putative gene-related peptides (fGRP-RP-1, -RP-2, and -RP-3). Subsequently, we identified these putative fGRP-RPs as mature peptides and analyzed their hypophysiotropic activities. Only fGRP-RP-2 stimulated the release of GH and prolactin (PRL) in vitro and in vivo. Thus, in addition to fGRP, fGRP-RP-2 acts as a hypothalamic factor on the frog pituitary to stimulate the release of GH and PRL.


Regulation of testicular function in the stallion: an intricate network of endocrine, paracrine and autocrine systems.
It is well established in many mammalian species, including the horse that normal testicular function is dependent upon a functional hypothalamic-pituitary-testicular (HPT) axis, which involves classic feedback mechanisms. The major HPT hormones involved in the stallion are gonadotropin-releasing hormone (GnRH), luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone (T), estrogens (Es) and inhibin (INH). Although prolactin (PRL) fluctuates with season in the stallion and both PRL and thyroid hormone (TH) affect reproduction in other male species, their effects on stallion reproduction have not been elucidated. Growth hormone (GH) in the stallion may be involved in sperm motility, production and secretion of insulin-like growth factor-1 (IGF-1) and LH-induced testosterone release. The action of these hormones and the products involved for normal spermatogenesis require cell to cell communication within the testis. The somatic cell types, Leydig, Sertoli and peritubular myoid cells, all support germ cell development, maturation and release into the seminiferous tubule lumen. The cell to cell crosstalk involves an intricate network of paracrine-autocrine systems that support the endocrine input to modulate cell function. In other male species, researchers have demonstrated the reproductive effects of such paracrine-autocrine factors as IGF-1, transferrin, androgens, estrogens, inhibin, insulin like peptide 3 (INSL3), beta-endorphin and oxytocin. The specific nature and relative contribution of these various factors on testicular function in fertile and subfertile stallions are under investigation. This review summarizes current information regarding the nature of the multiple endocrine-paracrine-autocrine systems that may be necessary for normal testicular function in the stallion.


Overground Walking With a Transparent Exoskeleton Shows Changes in Spatiotemporal Gait Parameters.
Lower-limb gait training (GT) exoskeletons have been successfully used in rehabilitation programs to overcome the burden of locomotor impairment. However, providing suitable net interaction torques to assist patient movements is still a challenge. Previous transparent operation approaches have been tested in treadmill-based GT exoskeletons to improve user-robot interaction. However, it is not yet clear how a transparent lower-limb GT system affects user's gait kinematics during overground walking, which unlike treadmill-based systems, requires active participation of the subjects to maintain stability. In this study, we implemented a transparent operation strategy on the ExoRoboWalker, an overground GT exoskeleton, to investigate its effect on the user's gait. The approach employs a feedback zero-torque controller with feedforward compensation for the exoskeleton's dynamics and actuators' impedance. We analyzed the data of five healthy subjects walking overground with the exoskeleton in transparent mode (ExoTransp) and non-transparent mode (ExoOff) and walking without exoskeleton (NoExo). The transparent controller reduced the user-robot interaction torque and improved the user's gait kinematics relative to ExoOff. No significant difference in stride length is observed between ExoTransp and NoExo (p = 0.129). However, the subjects showed a significant difference in cadence between ExoTransp (50.9± 1.1 steps/min) and NoExo (93.7 ± 8.7 steps/min) (p = 0.015), but not between ExoTransp and ExoOff (p = 0.644). Results suggest that subjects wearing the exoskeleton adjust their gait as in an attention-demanding task changing the spatiotemporal gait characteristics likely to improve gait balance.


Modeling the Longitudinal Effects of Insight on Depression, Quality of Life and Suicidality in Schizophrenia Spectrum Disorders: Results from the FACE-SZ Cohort.
Up to half of the patients with schizophrenia attempt suicide during their lifetime. Better insight is associated with better functioning but also with increased suicidality. The direction of the relationship between insight and suicidality is not clear, hence we aimed to provide new elements using structural equation modeling. Insight, quality of life (QoL), depression, and suicidality were measured at baseline and at 12 months in individuals with schizophrenia spectrum disorders. The relationships between these variables were investigated by latent difference score models, controlling for chlorpromazine doses, positive and negative symptoms, and general psychopathology. 738 patients were included, and 370 completed the study. Baseline levels of insight predicted changes in suicidality, whereas baseline levels of suicidality did not predict changes in insight, suggesting that better insight underlies suicidality and predicts its worsening. Our results suggest this temporal sequence: better insight → worse QoL → increased depression → increased suicidality, while insight also affects the three variables in parallel. Better insight predicts a worsening of QoL, depression and suicidality. These findings contribute to our global understanding of the longitudinal influence of insight on suicidality. We advocate that insight-targeted interventions should not be proposed without the monitoring of depression and suicide prevention.


What is all that thrombin for?
The hemostatic process initiated by the exposure of tissue factor to blood is a threshold limited reaction which occurs in two distinct phases. During an initiation phase, small amounts of factor (F)Xa, FIXa and thrombin are generated. The latter activates the procofactors FV and FVIII to the activated cofactors which together with their companion serine proteases form the intrinsic FX activator (FVIIIa-FIXa) and prothrombinase (FVa-FXa) which generate the bulk of FXa and thrombin during a propagation phase. The clotting process (fibrin formation) occurs at the inception of the propagation phase when only 5-10 nM thrombin has been produced. Consequently, the vast majority (greater than 95%) of thrombin is produced after clotting during the propagation phase of thrombin generation. The blood of individuals with either hemophilia A or hemophilia B has no ability to generate the intrinsic FXase, and hence is unable to support the propagation phase of the reaction. Since clot based assays conclude before the propagation phase they are not sensitive to hemophilia A and B. The inception and magnitude of the propagation phase of thrombin generation is influenced by genetic polymorphisms associated with thrombotic and hemorrhagic disease, by the natural abundance of pro- and anticoagulants in healthy individuals and by pharmacologic interventions which influence thrombotic pathology. Therefore, it is our suspicion that the performance of the entire process of thrombin generation from initiation through propagation and termination phases of the reaction are relevant with respect to both hemorrhagic and thrombotic pathology.


Interactions of pH and K+ conductance in cultured bovine retinal pigment epithelial cells.
Passive ion transport properties were studied in confluent monolayers of cultured bovine retinal pigment epithelial cells using intracellular microelectrode technique. The mean stable intracellular (designated by subscript i) potential was -59.1 +/- 0.8 (SE) mV. Extracellular (designated by subscript o) acidification induced a depolarization, whereas alkalinization induced a hyperpolarization. These effects were observed both in bicarbonate-free as well as in HCO3- Ringer (pHo changed by varying [HCO3-]o at constant pCO2). Acidification of pHi (changed by addition and removal of butyrate, CO2 or NH3) also caused a depolarization. Complete removal of HCO3-/CO2 at constant pHo caused a hyperpolarization. K+ transference, checked by applying high K+o, increased with K+o. It decreased with both extra and intracellular acidification and increased with alkalinization. In the presence of Ba2+, voltage reactions to changes in either pHo or pHi were greatly reduced. Depolarization by 40 mM K+ caused a similar reduction. It is suggested that K+ conductance of bovine retinal pigment epithelial cells is reduced by either intra- or extracellular acidification at normal [K+]o. Depolarization by high K+ induces an increase in K+ transference and reduces pH sensitivity.


Autosomal dominant osteosclerosis associated with familial spinal canal stenosis.
We studied a family with autosomal dominant osteosclerosis associated with familial spinal canal stenosis. The propositus, a 44-year-old Japanese woman, had a 9-month history of occipitalgia and left tinnitus, and also had a 2-month history of pain and numbness of the right upper limb. Radiographic skeletal survey showed osteosclerotic changes in the neurocranium, diaphysis of the long bone, mandible, shoulder, clavicle, and ribs. Serum alkaline phosphatase was normal, and no periosteal excrescences were seen. The inheritance pattern was autosomal dominant. The propositus and her daughter, both with severe osteosclerosis, showed spinal canal stenosis, but her son, whose osteosclerosis was moderate, did not. This is the first report of autosomal dominant osteosclerosis associated with familial spinal canal stenosis.


Pain assessment in animal models: do we need further studies?
In the last two decades, animal models have become important tools in understanding and treating pain, and in predicting analgesic efficacy. Although rodent models retain a dominant role in the study of pain mechanisms, large animal models may predict human biology and pharmacology in certain pain conditions more accurately. Taking into consideration the anatomical and physiological characteristics common to man and pigs (median body size, digestive apparatus, number, size, distribution and communication of vessels in dermal skin, epidermal-dermal junctions, the immunoreactivity of peptide nerve fibers, distribution of nociceptive and non-nociceptive fiber classes, and changes in axonal excitability), swines seem to provide the most suitable animal model for pain assessment. Locomotor function, clinical signs, and measurements (respiratory rate, heart rate, blood pressure, temperature, electromyography), behavior (bright/quiet, alert, responsive, depressed, unresponsive), plasma concentration of substance P and cortisol, vocalization, lameness, and axon reflex vasodilatation by laser Doppler imaging have been used to assess pain, but none of these evaluations have proved entirely satisfactory. It is necessary to identify new methods for evaluating pain in large animals (particularly pigs), because of their similarities to humans. This could lead to improved assessment of pain and improved analgesic treatment for both humans and laboratory animals.


Lessons to be learned from Pierre Fauchard.
Pierre Fauchard possessed the following attributes: Curiosity, intelligence, courage, perseverance, honesty, dexterity, and social responsibility. I suspect that these qualities would have assured him success in whatever career he might have followed. He chose surgery, and through his experience as a ship's doctor in the French navy, he specialized in diseases of the mouth and teeth (common among seamen), qualifying as a surgeon-dentist prior to establishing his highly successful private practice in Paris. In his long career he stood for excellence, the scientific approach, comprehensive care, high ethical standards, and technical innovation. Although we know almost nothing of his private life, his book does reveal a good part of his character. Based solely on this source it is possible to conclude that Fauchard was a true son of the enlightenment, who justifiably holds the title of Father of Modern Dentistry. His treatise contains lessons to be emulated even into the 21st century.


Targeted activation of RNA helicase retinoic acid-inducible gene-I induces proimmunogenic apoptosis of human ovarian cancer cells.
Most malignant cells are poorly immunogenic and fail to elicit an effective antitumor immune response. In contrast, viral infections of cells are promptly detected and eliminated by the immune system. Viral recognition critically hinges on cytosolic nucleic acid receptors that include the proinflammatory RNA helicase retinoic acid-inducible gene-I (RIG-I). Here, we show that targeted delivery of RIG-I agonists induced ovarian cancer cells to upregulate HLA class I and to secrete the proinflammatory cytokines CXCL10, CCL5, interleukin-6, tumor necrosis factor-alpha, and IFN-beta. Ovarian cancer cells stimulated via RIG-I became apoptotic and were readily phagocytosed by monocytes and monocyte-derived dendritic cells, which in turn upregulated HLA class I/II and costimulatory molecules and released CXCL10 and IFN-alpha. Our findings offer proof of principle that mimicking viral infection in ovarian cancer cells triggers an immunogenic form of tumor cell apoptosis that may enhance immunotherapy of ovarian cancer.


Impact of the indexed effective orifice area on mid-term cardiac-related mortality after aortic valve replacement.
There has been ongoing controversy as to whether prosthesis-patient mismatch (PPM, defined as indexed effective orifice area (EOAI) <0.85 m(2)/cm(2)) influences mortality after aortic valve replacement (AVR). In most studies, PPM is anticipated by reference tables based on mean EOAs as opposed to individual assessment. These reference values may not reflect the actual in vivo EOAI and hence, the presence or absence of PPM may be based on false assumptions. To assess the impact of small prosthesis EOA on survival after aortic valve replacement AVR. 645 patients had undergone an AVR between 2000 and 2007 entered the study. All patients underwent transthoracic echocardiography for determination of the actual EOAI within 6 months postoperatively. In order to predict time from surgery to death a proportional hazards model for competing risks (cardiac death vs death from other causes) was used. EOAI was entered as a continuous variable. PPM occurred in 40% of the patients. After a median follow-up of 2.35 years, 92.1% of the patients were alive. The final Cox regression model showed a significantly increased risk for cardiac death among patients with a smaller EOAI (HR=0.32, p=0.022). The effect of EOAI on the 2-5 year mortality risk was demonstrated by risk plots. In contrast to previous studies these EOAI values were obtained through postoperative echocardiography, substantially improving the accuracy of measurement, and the EOAI was modelled as a continuous variable. There was a significantly improved survival for larger EOAIs following AVR. Strategies to avoid PPM should become paramount during AVR.


Cortical silent period duration and its implications for surround inhibition of a hand muscle.
Surround inhibition is a neural mechanism that assists in the focusing of excitatory drive to muscles responsible for a given movement (agonist muscles) by suppressing unwanted activity in muscles not relevant to the movement (surround muscles). The purpose of the study was to determine the contribution of γ-aminobutyric acid(B) receptor-mediated intracortical inhibition, as assessed by the cortical silent period (CSP), to the generation of surround inhibition in the motor system. Eight healthy adults (five women and three men, 29.8 ± 9 years) performed isometric contractions with the abductor digiti minimi (ADM) muscle in separate conditions with and without an index finger flexion movement. The ADM motor evoked potential amplitude and CSP duration elicited by transcranial magnetic stimulation were compared between a control condition in which the ADM was activated independently and during conditions involving three phases (pre-motor, phasic, and tonic) of the index finger flexion movement. The motor evoked potential amplitude of the ADM was greater during the control condition compared with the phasic condition. Thus, the presence of surround inhibition was confirmed in the present study. Most critically, the CSP duration of the ADM decreased during the phasic stage of finger flexion compared with the control condition, which indicated a reduction of this type of intracortical inhibition during the phasic condition. These findings indicate that γ-aminobutyric acid(B) receptor-mediated intracortical inhibition, as measured by the duration of the CSP, does not contribute to the generation of surround inhibition in hand muscles.


Information environments for supporting consistent registrar medical handover.
This study was two-fold in nature. Initially, it examined the information environment and the use of customary information tools to support medical handovers in a large metropolitan teaching hospital on four weekends (i.e. Friday night to Monday morning). Weekend medical handovers were found to involve sequences of handovers where patients were discussed at the discretion of the doctor handing over; no reliable discussion of all patients of concern occurred at any one handover, with few information tools being used; and after a set of weekend handovers, there was no complete picture on a Monday morning without an analysis of all patient progress notes. In a subsequent case study, three information tools specifically designed as intervention that attempted to enrich the information environment were evaluated. Results indicate that these tools did support greater continuity in who was discussed but not in what was discussed at handover. After the intervention, if a doctor discussed a patient at handover, that patient was more likely to be discussed at subsequent handovers. However, the picture at Monday morning remained fragmentary. The results are discussed in terms of the complexities inherent in the handover process.


Rapid fluorescent detection of (anti)androgens with spiggin-gfp medaka.
Widespread environmental antiandrogen contamination has been associated with negative impacts on biodiversity and human health. In particular, many pesticides are antiandrogenic, creating a need for robust and sensitive environmental monitoring. Our aim was to develop a sensitive and specific transgenic medaka (Oryzias latipes) model bearing an androgen responsive fluorescent reporter construct for whole organism-based environmental screening of pro- and antiandrogens. We analyzed the 5' regions of the androgen responsive three-spined stickleback (Gasterosteus aculeatus) spiggin genes in silico, revealing conserved blocks of sequence harboring androgen response elements. Identified putative promoters were cloned upstream of GFP. Germinal transgenesis with spg1-gfp led to stable medaka lines. GFP induction was exclusive to the kidney, the site of spiggin protein production in sticklebacks. Significant GFP expression was induced by three or four-day androgen treatment of newly hatched fry, but not by estrogens, mineralocorticoids, glucocorticoids or progestogens. The model responded dose-dependently to androgens, with highest sensitivity to 17MT (1.5 μg/L). In addition to flutamide, the biocides fenitrothion, vinclozolin and linuron significantly inhibited 17MT-induced GFP induction, validating the model for detection of antiandrogens. The spg1-gfp medaka model provides a sensitive, specific, and physiologically pertinent biosensor system for analyzing environmental androgen activity.


CFAP300 mutation causing primary ciliary dyskinesia in Finland.
Primary ciliary dyskinesia (PCD) is a rare genetic condition characterized by chronic respiratory tract infections and in some cases laterality defects and infertility. The symptoms of PCD are caused by malfunction of motile cilia, hair-like organelles protruding out of the cell that are responsible for removal of mucus from the airways and organizing internal organ positioning during embryonic development. PCD is caused by mutations in genes coding for structural or assembly proteins in motile cilia. Thus far mutations in over 50 genes have been identified and these variants explain around 70% of all known cases. Population specific genetics underlying PCD has been reported, thus highlighting the importance of characterizing gene variants in different populations for development of gene-based diagnostics. In this study, we identified a recurrent loss-of-function mutation c.198_200delinsCC in CFAP300 causing lack of the protein product. PCD patients homozygous for the identified CFAP300 mutation have immotile airway epithelial cilia associated with missing dynein arms in their ciliary axonemes. Furthermore, using super resolution microscopy we demonstrate that CFAP300 is transported along cilia in normal human airway epithelial cells suggesting a role for CFAP300 in dynein complex transport in addition to preassembly in the cytoplasm. Our results highlight the importance of CFAP300 in dynein arm assembly and improve diagnostics of PCD in Finland.


New graduate nurses' perceptions of the effects of clinical simulation on their critical thinking, learning, and confidence.
Critical thinking has been a crucial outcome of nursing educational programs. Effective nurses should be knowledgeable about complex patient situations and confident in their skills. One teaching strategy recently adopted by some educators to develop nurses' critical thinking, learning, and confidence is simulation. Simulation incorporates scenarios and case studies developed to replicate real-life clinical situations. Learners are asked to solve clinical problems and make critical decisions based on the information provided. Little research has been done on how simulation experiences promote critical thinking, learning, and confidence, especially in new graduate nurses. This study explored the perceptions of new graduate nurses of how clinical simulation developed their critical thinking skills, learning, and confidence throughout their hospital clinical training. Ten new baccalaureate nursing graduates voluntarily participated in this study, which used an exploratory descriptive design. Data were collected by demographic questionnaires and semi-structured interviews and were analyzed using content analysis. Participants reported that simulation prepared them well to care confidently for critically ill patients. Simulation also helped them learn to make sound clinical decisions to improve patient outcomes. The findings have crucial implications for nursing education, practice, and research. They provide evidence to support the use of simulation as a teaching strategy to promote critical thinking skills, learning, and confidence.


[Role of the tumor suppressor ARF in oncogenesis].
The paper reviews the data available in the literature on a role of the tumor suppressor ARF in oncogenesis and considers the structure of a gene encoding ARF protein. The p53-dependent and p-53-independent functions of this protein are under many studies. There is evidence for the implication of ARF in angiogenesis. There is more and more information on the role of ARF in the regulation of a cell cycle, apoptosis, and autophagy. The importance of this tumor suppressor in the mechanisms of carcinogenesis is beyond question as the inactivation of ARF suppressor activity leads to the rapid growth of neoplasia. However, the exact mechanisms of ARF action yet remain unclear and require further studies by different specialists at both the molecular genetic and other levels of investigation.


Yeast transfer RNA: a small-angle x-ray study.
The intensity of x-ray scattering and the radius of gyration were measured for a mixture of yeast transfer RNA's in tenth molar potassium chloride. The experimentally observed radius of gyration eliminates single-stranded, hairpin, singly folded hairpin, triple-stranded, and linked double-hairpin models of tRNA but allows certain folded cloverleaf models. The measured intensities at larger angles, beyond the radius of gyration region, lend some support to the Holley's cloverleaf model, in which three arms are folded up tightly together and the fourth arm is extended in the opposite direction.


Phytochemical and biological activity studies of the Bhutanese medicinal plant Corydalis crispa.
The chemical constituents and biological activities of Corydalis crispa (Fumariaceae) were investigated for the first time. The phytochemical study resulted in the isolation of nine known isoquinoline alkaloids: protopine (1), 13-oxoprotopine (2), 13-oxocryptopine (3), stylopine (4), coreximine (5), rheagenine (6), ochrobirine (7), sibiricine (8) and bicuculline (9), with complete NMR data for 2 and 3 provided here for the first time. Crude extracts exhibitedsignificant anti-inflammatory (p < 0.01) activity against TNF-alpha production in LPS activated THP-1 cells. The acetylcholinesterase inhibitory activity of compounds 2, 4 and 7 and the antiplasmodial activity of compound 5 against P. falciparum strains TM4/8.2 and K1CB1 (multidrug resistant strain) are reported here for the first time. Stylopine (4) did not show antimalarial activity against the K1CB1 strain in contrast to a previous report. This study generated a scientific basis for the use of this plant in Bhutanese traditional medicine, either individually or in combination with other medicinal ingredients to treat a broad range of disorders. This study also identified compound 5 as potential new antimalarial lead compound.


Secretion of prebeta HDL increases with the suppression of cholesteryl ester transfer protein in Hep G2 cells.
Prebeta HDL are small, protein rich lipoproteins that are predominantly composed of apo A-I, without apo A-II. Prebeta HDL are secreted from the liver as nascent HDL and/or are produced in the incubated plasma by cholesteryl ester transfer protein (CETP). However, the role of CETP in the secretion of HDL from the liver has yet to be determined. In the present study, we examined the effect of the suppression of hepatic CETP by antisense oligodeoxynucleotides (ODNs) against CETP targeted to the liver on the secretion of apo A-I using a Hep G2 cell culture. The ODNs against CETP were coupled to asialoglycoprotein (ASOR) carrier molecules, which serve as an important method for the regulation of liver gene expression. Hep G2 cells were cultured in DMEM supplemented with 10 FBS. After 2 days, the medium was changed to DMEM with EGF and the cells were divided into three groups. The control group received saline, while the sense group was mixed with the sense ODNs complex and the antisense group was mixed with the antisense ODNs complex, respectively, for 2 days. Both the hepatic CETP mRNA and the CETP mass in the medium in the antisense group decreased significantly more than in the sense and the control groups (CETP mass: 1.697 + /- 0.410 ng/mg cell protein vs. 2.367 + /- 0.22 and 2.360 + /- 0.139, n = 3 in each determination). In contrast, both the hepatic apo A-I mRNA and the apo A-I mass in the medium in the antisense group were significantly higher than those in the sense and the control groups (apo A-I mass; 1.877 + /- 0.215 micro/mg cell protein vs. 1.213 + /- 0.282 and 1.097 + /- 0.144, n = 3 in each determination). The increase in apo A-I was mainly due to the increase in prebeta apo A-I. These findings may partly explain why HDL and apo A-I increase in patients with CETP deficiency, while also indicating the possibility that the original level of prebeta HDL is sufficient in such patients.


Osmotic regulation of transcription: induction of the proU betaine transport gene is dependent on accumulation of intracellular potassium.
The proU locus, which encodes a high-affinity betaine transport system, and the kdp operon, which encodes a potassium transport system, are the principal osmoresponsive genes in Escherichia coli and Salmonella typhimurium. The kdp operon is known to be induced in response to changes in cell turgor. We have investigated the control of proU expression and shown that it differs from that of kdp in a number of fundamental ways. Rather than responding to changes in turgor, proU expression is principally determined by the intracellular accumulation of potassium ions. Potassium and betaine were shown to play distinct osmoprotective roles. Potassium serves as the principal osmoprotectant and is accumulated in response to low-level osmotic stress to restore turgor. As external osmolarity is increased to a level at which the corresponding increase in internal potassium concentrations is potentially deleterious to enzyme function, betaine (when available) is accumulated in preference to potassium. The different mechanisms of proU and kdp regulation reflect the different physiological roles of these two osmoprotectants.


Brain atrophy in dementia judged by CT scan ranking.
In order to further investigate the relationships of brain atrophy, age, and dementia, computed tomographic (CT) scans on two groups of subjects aged 65-80 years were rank-ordered according to ventricular and sulcal size. Each group contained subjects with mild senile dementia of the Alzheimer type (SDAT) and matched controls. In both experiments there was a strong correlation between these rank orders, ventricular size, and the clinical dementia rating. There was also a weaker but significant correlation between rank ordering and sulcal size. This suggests that brain atrophy in excess of that normal for age alone occurs in early SDAT.


Natural and Recombinant SARS-CoV-2 Isolates Rapidly Evolve In Vitro to Higher Infectivity through More Efficient Binding to Heparan Sulfate and Reduced S1/S2 Cleavage.
One of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virulence factors is the ability to interact with high affinity to the ACE2 receptor, which mediates viral entry into cells. The results of our study demonstrate that within a few passages in cell culture, both the natural isolate of SARS-CoV-2 and the recombinant cDNA-derived variant acquire an additional ability to bind to heparan sulfate (HS). This promotes a primary attachment of viral particles to cells before their further interactions with the ACE2. Interaction with HS is acquired through multiple mechanisms. These include (i) accumulation of point mutations in the N-terminal domain (NTD) of the S protein, which increases the positive charge of the surface of this domain, (ii) insertions into the NTD of heterologous peptides containing positively charged amino acids, and (iii) mutation of the first amino acid downstream of the furin cleavage site. This last mutation affects S protein processing, transforms the unprocessed furin cleavage site into the heparin-binding peptide, and makes viruses less capable of syncytium formation. These viral adaptations result in higher affinity of viral particles to heparin, dramatic increase in plaque sizes, more efficient viral spread, higher infectious titers, and 2 orders of magnitude higher infectivity. The detected adaptations also suggest an active role of NTD in virus attachment and entry. As in the case of other RNA-positive (RNA+) viruses, evolution to HS binding may result in virus attenuation in vivo. IMPORTANCE The spike protein of SARS-CoV-2 is a major determinant of viral pathogenesis. It mediates binding to the ACE2 receptor and, later, fusion of viral envelope and cellular membranes. The results of our study demonstrate that SARS-CoV-2 rapidly evolves during propagation in cultured cells. Its spike protein acquires mutations in the NTD and in the P1' position of the furin cleavage site (FCS). The amino acid substitutions or insertions of short peptides in NTD are closely located on the protein surface and increase its positive charge. They strongly increase affinity of the virus to heparan sulfate, make it dramatically more infectious for the cultured cells, and decrease the genome equivalent to PFU (GE/PFU) ratio by orders of magnitude. The S686G mutation also transforms the FCS into the heparin-binding peptide. Thus, the evolved SARS-CoV-2 variants efficiently use glycosaminoglycans on the cell surface for primary attachment before the high-affinity interaction of the spikes with the ACE2 receptor.


Morphological image analysis of quantum motion in billiards.
Morphological image analysis is applied to the time evolution of the probability distribution of a quantum particle moving in two- and three-dimensional billiards. It is shown that the time-averaged Euler characteristic of the probability distribution provides a well defined quantity to distinguish between classically integrable and nonintegrable billiards. In three dimensions the time-averaged mean breadth of the probability distribution may also be used for this purpose.


Release of the National Scheme's Aboriginal and Torres Strait Islander Health and Cultural Safety Strategy 2020-2025; the impacts for podiatry in Australia: a commentary.
Developing since colonisation, Australia's healthcare system has dismissed an ongoing and successful First Nations health paradigm in place for 60,000 years. From Captain James Cook documenting 'very old' First Nations Peoples being 'far more happier than we Europeans' and Governor Arthur Phillip naming Manly in admiration of the physical health of Gadigal men of the Eora Nation, to anthropologist Daisy Bates' observation of First Nations Peoples living 'into their eighties' and having a higher life expectancy than Europeans; our healthcare system's shameful cultural safety deficit has allowed for an Aboriginal and Torres Strait Islander child born in Australia today to expect to live 9 years less than a non-Indigenous child. Disproportionately negative healthcare outcomes including early onset diabetes-related foot disease and high rates of lower limb amputation in Aboriginal and Torres Strait Islander Peoples contribute to this gross inequity. In 2020, the Australian Health Practitioner Regulation Authority released the National Scheme's Aboriginal and Torres Strait Islander Health and Cultural Safety Strategy 2020-2025 - empowering all registered health practitioners within Australia to provide health care to Aboriginal and Torres Strait Islander Peoples that is inclusive, respectful and safe, as judged by the recipient of care. This recently released strategy is critically important to the podiatry profession in Australia. As clinicians, researchers and educators we have a collective responsibility to engage with this strategy of cultural safety. This commentary defines cultural safety for podiatry and outlines the components of the strategy in the context of our profession. Discussion considers the impact of the strategy on podiatry. It identifies mechanisms for podiatrists in all settings to facilitate safer practice, thereby advancing healthcare to produce more equitable outcomes. Aboriginal and Torres Strait Islander Peoples access health services more frequently and have better health outcomes where provision of care is culturally safe. By engaging with the National Scheme's Aboriginal and Torres Strait Islander Health and Cultural Safety Strategy, all registered podiatrists in Australia can contribute to achieving equity in health outcomes for Aboriginal and Torres Strait Islander Peoples.


Human Cytomegalovirus Interleukin 10 Homologs: Facing the Immune System.
Human Cytomegalovirus (HCMV) can cause a variety of health disorders that can lead to death in immunocompromised individuals and neonates. The HCMV lifecycle comprises both a lytic (productive) and a latent (non-productive) phase. HCMV lytic infection occurs in a wide range of terminally differentiated cell types. HCMV latency has been less well-studied, but one characterized site of latency is in precursor cells of the myeloid lineage. All known viral genes are expressed during a lytic infection and a subset of these are also transcribed during latency. The UL111A gene which encodes the viral IL-10, a homolog of the human IL-10, is one of these genes. During infection, different transcript isoforms of UL111A are generated by alternative splicing. The most studied of the UL111A isoforms are cmvIL-10 (also termed the "A" transcript) and LAcmvIL-10 (also termed the "B" transcript), the latter being a well-characterized latency associated transcript. Both isoforms can downregulate MHC class II, however they differ in a number of other immunomodulatory properties, such as the ability to bind the IL10 receptor and induce signaling through STAT3. There are also a number of other isoforms which have been identified which are expressed by differential splicing during lytic infection termed C, D, E, F, and G, although these have been less extensively studied. HCMV uses the viral IL-10 proteins to manipulate the immune system during lytic and latent phases of infection. In this review, we will discuss the literature on the viral IL-10 transcripts identified to date, their encoded proteins and the structures of these proteins as well as the functional properties of all the different isoforms of viral IL-10.


The P2Y-like receptor GPR17 as a sensor of damage and a new potential target in spinal cord injury.
Upon central nervous system injury, the extracellular concentrations of nucleotides and cysteinyl-leukotrienes, two unrelated families of endogenous signalling molecules, are markedly increased at the site of damage, suggesting that they may act as 'danger signals' to alert responses to tissue damage and start repair. Here we show that, in non-injured spinal cord parenchyma, GPR17, a P2Y-like receptor responding to both uracil nucleotides (e.g. UDP-glucose) and cysteinyl-leukotrienes (e.g. LTD4 and LTC4), is present on a subset of neurons and of oligodendrocytes at different stages of maturation, whereas it is not expressed by astrocytes. GPR17 immunoreactivity was also found on ependymal cells lining the central canal that still retain some of the characteristics of stem/progenitor cells during adulthood. Induction of spinal cord injury (SCI) by acute compression resulted in marked cell death of GPR17+ neurons and oligodendrocytes inside the lesion followed by the appearance of proliferating GPR17+ microglia/macrophages migrating to and infiltrating into the lesioned area. Moreover, 72 h after SCI, GPR17+ ependymal cells started to proliferate and to express GFAP, suggesting their activation and 'de-differentiation' to pluripotent progenitor cells. The in vivo knock down of GPR17 by an antisense oligonucleotide strategy during SCI induction markedly reduced tissue damage and related histological and motor deficits, thus confirming the crucial role played by this receptor in the early phases of tissue damage development. Taken together, our findings suggest a dual and spatiotemporal-dependent role for GPR17 in SCI. At very early times after injury, GPR17 mediates neuronal and oligodendrocyte death inside the lesioned area. At later times, GPR17+ microglia/macrophages are recruited from distal parenchymal areas and move toward the lesioned zone, to suggest a role in orchestrating local remodelling responses. At the same time, the induction of the stem cell marker GFAP in GPR17+ ependymal cells suggests initiation of repair mechanisms. Thus, GPR17 may act as a 'sensor' of damage that is activated by nucleotides and cysteinyl-leukotrienes released in the lesioned area, and could also participate in post-injury responses. Moreover, its presence on spinal cord pre-oligodendrocytes and precursor-like cells suggests GPR17 as a novel target for therapeutic manipulation to foster remyelination and functional repair in SCI.


The collagen V homotrimer [alpha1(V)](3) production is unexpectedly favored over the heterotrimer [alpha1(V)](2)alpha2(V) in recombinant expression systems.
Collagen V, a fibrillar collagen with important functions in tissues, assembles into distinct chain associations. The most abundant and ubiquitous molecular form is the heterotrimer [alpha1(V)](2)alpha2(V). In the attempt to produce high levels of recombinant collagen V heterotrimer for biomedical device uses, and to identify key factors that drive heterotrimeric chain association, several cell expression systems (yeast, insect, and mammalian cells) have been assayed by cotransfecting the human proalpha1(V) and proalpha2(V) chain cDNAs. Suprisingly, in all recombinant expression systems, the formation of [alpha1(V)](3) homotrimers was considerably favored over the heterotrimer. In addition, pepsin-sensitive proalpha2(V) chains were found in HEK-293 cell media indicating that these cells lack quality control proteins preventing collagen monomer secretion. Additional transfection with Hsp47 cDNA, encoding the collagen-specific chaperone Hsp47, did not increase heterotrimer production. Double immunofluorescence with antibodies against collagen V alpha-chains showed that, contrary to fibroblasts, collagen V alpha-chains did not colocalized intracellularly in transfected cells. Monensin treatment had no effect on the heterotrimer production. The heterotrimer production seems to require specific machinery proteins, which are not endogenously expressed in the expression systems. The different constructs and transfected cells we have generated represent useful tools to further investigate the mechanisms of collagen trimer assembly.


A comparitive study to evaluate the role of serum lipid levels in aetiology of carcinoma breast.
Breast cancer is the most common occurring cancer in women worldwide. Various factors that have lead to change in lifestyle are thought to be associated with increase in breast cancer incidence in Asian countries. Several clinical studies suggest the prognostic significance of serum lipid levels in breast cancer. Present study was planned to confirm the association of lipids levels with breast cancer. Lipid analyses was carried out on serum samples from 100 breast cancer patients and 100 control women, with a age group of 25 y and above. The case & control group included 50 premenopausal women and 50 postmenopausal women. Patients of study group had high Low density lipoproteins (LDL) (p-value - 0.00), Triglycerides (TG) (p-value -0.001) and Total cholesterol (TC) (p-value -0.00) as compared to control group. There was no association of VLDL (p-value -0.436) and HDL (p-value -0.797) among study group and control group. There was no association of lipid levels in different grades of carcinoma. High levels of LDL, TG and TC were associated with breast cancer. However, no association was there in different grades of carcinoma. So it may be concluded that, LDL, TG and TC may be having some role in aetiology of breast cancer. Therefore, early detection and control of these factors may help in reducing the incidence of breast cancer and ultimately social and economic burden on society.


Electrospray ionization (ESI) fragmentations and dimethyldioxirane reactivities of three diverse lactams having full, half, and zero resonance energies.
Three lactams having, respectively, ~20, ~10, and 0 kcal/mol of resonance energy have been subjected to electrospray ionization mass spectrometry (ESI/MS) as well as to attempted reaction with dimethyldioxirane (DMDO). The ESI/MS for all three lactams are consistent with fragmentation from the N-protonated, rather than the O-protonated tautomer. Each exhibits a unique fragmentation pathway. DFT calculations are employed to provide insights concerning these pathways. N-Ethyl-2-pyrrolidinone and 1-azabicyclo[3.3.1]nonan-2-one, the full- and half-resonance lactams, are unreactive with DMDO. The "Kirby lactam" (3,5,7-trimethyl-1-azaadamantan-2-one) has zero resonance energy and reacts rapidly with DMDO to generate a mixture of reaction products. The structure assigned to one of these is the 2,2-dihydroxy-N-oxide, thought to be stabilized by intramolecular hydrogen bonding and buttressing by the methyl substituents. A reasonable pathway to this derivative might involve formation of an extremely labile N-oxide, in a purely formal sense, an example of the hitherto-unknown amide N-oxides, followed by hydration with traces of moisture.


Impact of concentrations of camphor on the in vitro mixed ruminal microorganism fermentation from goats selected for consumption of low and high levels of Juniperus spp.-2.
The microbial population in the gastrointestinal tract of ruminant animals aids in the utilization of forages with high levels of secondary plant compounds. Two divergent bloodlines of meat goats have been selected by screening fecal samples with near-infrared reflectance spectroscopy to assess the goat's consumption of high or low levels of Juniperus sp. leaves containing several monoterpenes, including camphor. The mechanism by which these goats can consume greater concentrations of Juniperus spp. leaves than their counterparts is unclear, and therefore, this study was designed to determine if differences existed between the ruminal microbial populations of the low and high juniper-consuming bloodlines (LJC vs. HJC) by analyzing their ruminal microbiota and fermentation end products. In the present study, concentrations (0.00, 0.5, 0.99, 1.97, or 5.91 mM) of camphor were added to mixed ruminal microorganism fermentation. Five LJC and five HJC goats were fed a juniper-free diet (n = 10), and five LJC and five HJC goats (n = 10) were fed a diet that contained 30% fresh Juniperus ashei leaves for 21 d prior to ruminal fluid collection. In vitro fermentations used LJC and HJC, ruminal fluid inoculum added (33% v/v) to anoxic media in sealed Balch tubes. Camphor increased (P < 0.05) total short-chain fatty acid (SCFA) concentrations for all but one experimental group. Between the main dietary and bloodline goat effects, the diet was significant for all SCFA results except butyrate. In contrast, bloodline was only significant for acetate and butyrate molar proportions. Rumen fluid from juniper-free-fed goats exhibited greater concentrations of Ruminococcaceae, whereas juniper-fed goats contained more Coriobacteriaceae. Results demonstrated that mixed ruminal microorganisms fermentations from HJC goats did not produce greater concentrations of SCFAs or have the ability to degrade camphor at a higher rate than did that from LJC goats. Results suggest that camphor tolerance from J. ashei, was related to hepatic catabolic mechanisms instead of ruminal microbial degradation; however, further in vivo work is warranted.


Lyme disease -- United States, 1996.
Lyme disease (LD) is caused by the tickborne spirochete Borrelia burgdorferi sensu lato and is the most common vectorborne disease in the United States. Surveillance for LD was initiated by CDC in 1982, and the Council of State and Territorial Epidemiologists designated it a nationally notifiable disease in January 1991. For surveillance purposes, LD is defined as the presence of an erythema migrans rash > or =5 cm in diameter or laboratory confirmation of infection with evidence of at least one manifestation of musculoskeletal, neurologic, or cardiovascular disease (1). This report summarizes the provisional number of cases of LD reported to CDC during 1996 and indicates that the number of cases reported to CDC was a record high.


Comparison of coronary revascularization procedures in octogenarians: a systematic review and meta-analysis.
Elderly patients are the fastest growing population in the US healthcare system and more patients aged 80 years and older require CABG or percutaneous coronary intervention (PCI) for coronary revascularization than ever before. Because octogenarian patients have not been adequately represented in randomized trials comparing CABG and PCI, the most appropriate method of revascularization for this group of patients has not been determined. We performed a systematic review and a meta-analysis of 66 studies of coronary revascularization in patients aged over 80 years. The primary endpoints included 30 day mortality and long-term survival. Subgroup analyses stratified by revascularization type (PCI versus CABG) were also performed. Pooled estimate of 30 day mortality was 6.3% (95% CI 5.3%-7.5%), and for survival at 1, 3 and 5 years, 86% (84%-88%), 78% (74%-81%) and 67% (61%-72%), respectively. A greater number of men (P <0.001) and patients with multivessel disease (P = 0.004) were treated with CABG than with PCI. Pooled estimates, based on type of revascularization, of 30 day mortality and 1 year survival were similar (7.3% [6.3%-8.2%] for CABG vs 5.4% [4.4%-6.4%] for PCI and 86% [83%-88%] for CABG vs 87% [84%-91%] for PCI, respectively). Available data indicate that revascularization can be performed in octogenarians with acceptable short-term and long-term outcomes; most of the evidence is, however, low level. Furthermore, it is unclear whether octogenarians derive greater survival benefit from CABG or from PCI because preprocedural risk profiles differ between intervention types. Periprocedural and long-term outcomes are, however, equivalent, and randomized, controlled trials of high-risk octogenarians are needed.


Upscaling Participatory Action and Videos for Agriculture and Nutrition (UPAVAN) trial comparing three variants of a nutrition-sensitive agricultural extension intervention to improve maternal and child nutritional outcomes in rural Odisha, India: study protocol for a cluster randomised controlled trial.
Maternal and child undernutrition have adverse consequences for pregnancy outcomes and child morbidity and mortality, and they are associated with low educational attainment, economic productivity as an adult, and human wellbeing. 'Nutrition-sensitive' agriculture programs could tackle the underlying causes of undernutrition. This study is a four-arm cluster randomised controlled trial in Odisha, India. Interventions are as follows: (1) an agricultural extension platform of women's groups viewing and discussing videos on nutrition-sensitive agriculture (NSA) practices, and follow-up visits to women at home to encourage the adoption of new practices shown in the videos; (2) women's groups viewing and discussing videos on NSA and nutrition-specific practices, with follow-up visits; and (3) women's groups viewing and discussing videos on NSA and nutrition-specific practices combined with a cycle of Participatory Learning and Action meetings, with follow-up visits. All arms, including the control, receive basic nutrition training from government community frontline workers. Primary outcomes, assessed at baseline and 32 months after the start of the interventions, are (1) percentage of children aged 6-23 months consuming ≥ 4 out of 7 food groups per day and (2) mean body mass index (BMI) (kg/m2) of non-pregnant, non-postpartum (gave birth > 42 days ago) mothers or female primary caregivers of children aged 0-23 months. Secondary outcomes are percentage of mothers consuming ≥ 5 out of 10 food groups per day and percentage of children's weight-for-height z-score < -2 standard deviations (SD). The unit of randomisation is a cluster, defined as one or more villages with a combined minimum population of 800 residents. There are 37 clusters per arm, and outcomes will be assessed in an average of 32 eligible households per cluster. For randomisation, clusters are stratified by distance to nearest town (< 10 km or ≥ 10 km), and low (< 30%), medium (30-70%), or high (> 70%) proportion of Scheduled Tribe or Scheduled Caste (disadvantaged) households. A process evaluation will assess the quality of implementation and mechanisms behind the intervention effects. A cost-consequence analysis will compare incremental costs and outcomes of the interventions. This trial will contribute evidence on the impacts of NSA extension through participatory, low-cost, video-based approaches on maternal and child nutrition and on whether integration with nutrition-specific goals and enhanced participatory approaches can increase these impacts. ISRCTN , ISRCTN65922679 . Registered on 21 December 2016.


Mid-sagittal plane detection for advanced physiological measurements in brain scans.
The process of diagnosing many neurodegenerative diseases, such as Parkinson's and progressive supranuclear palsy, involves the study of brain magnetic resonance imaging (MRI) scans in order to identify and locate morphological markers that can highlight the health status of the subject. A fundamental step in the pre-processing and analysis of MRI scans is the identification of the mid-sagittal plane, which corresponds to the mid-brain and allows a coordinate reference system for the whole MRI scan set. To improve the identification of the mid-sagittal plane we have developed an algorithm in Matlab® based on the k-means clustering function. The results have been compared with the evaluation of four experts who manually identified the mid-sagittal plane and whose performances have been combined with a cognitive decisional algorithm in order to define a gold standard. The comparison provided a mean percentage error of 1.84%. To further refine the automatic procedure we trained a machine learning system using the results from the proposed algorithm and the gold standard. We tested this machine learning system and obtained results comparable to medical raters with a mean absolute error of 1.86 slices. The system is promising and could be directly incorporated into broader diagnostic support systems.


How reliable are the results from functional magnetic resonance imaging?
Functional magnetic resonance imaging (fMRI) is one of the most important methods for in vivo investigation of cognitive processes in the human brain. Within the last two decades, an explosion of research has emerged using fMRI, revealing the underpinnings of everything from motor and sensory processes to the foundations of social cognition. While these results have revealed the potential of neuroimaging, important questions regarding the reliability of these results remain unanswered. In this paper, we take a close look at what is currently known about the reliability of fMRI findings. First, we examine the many factors that influence the quality of acquired fMRI data. We also conduct a review of the existing literature to determine if some measure of agreement has emerged regarding the reliability of fMRI. Finally, we provide commentary on ways to improve fMRI reliability and what questions remain unanswered. Reliability is the foundation on which scientific investigation is based. How reliable are the results from fMRI?


Precise Regulation of Juvenile Hormone III R-Stereoisomer Synthesis by Apis mellifera through Specifically Binding Methyl-(2E,6E)-farnesoate and Strictly Controlling Its Titer.
Juvenile hormone III (JH III) is a crucial hormone synthesized exclusively as R-stereoisomer in most insects. Herein, we established a mature Tris-HCl culture system for essential biochemical reactions and applied stable instrumental detection methods to analyze JH III, methyl farnesoate (MF) and juvenile hormone acid (JHA) using UPLC-MS/MS. Our results revealed that the R-JH III terminal synthesis pathway in Apis mellifera follows the "esterify then epoxidize" sequence, with precise methyl-(2E,6E)-farnesoate titer regulation and its spatial cis-trans isomerism, achieving selective R-JH III synthesis. Furthermore, we observed that the preferred generation of S/R-JH III chiral enantiomers varied depending on the spatial cis-trans isomerism of different MFs. Our results suggest that S-JH III could theoretically exist in insects, offering a novel perspective for understanding the synthesis mechanism of diverse complex juvenile hormones in different insect species.


Prevalence of central sleep apnea in people with tetraplegic spinal cord injury: a retrospective analysis of research and clinical data.
Over 80% of people with tetraplegia have sleep-disordered breathing (SDB), but whether this is predominantly obstructive or central is unclear. This study aimed to estimate the prevalence of central sleep apnea (CSA) in tetraplegia and the contributions of central, obstructive, and hypopnea respiratory events to SDB summary indices in tetraplegia. Research and clinical data from 606 individuals with tetraplegia and full overnight polysomnography were collated. The proportions of different respiratory event types were calculated; overall and for mild, moderate, and severe disease. The prevalence of Predominant CSA (Central Apnea Index [CAI] ≥ 5 and more central than obstructive apneas) and Any CSA (CAI ≥ 5) was estimated. Prevalence of sleep-related hypoventilation (SRH) was estimated in a clinical sub-cohort. Respiratory events were primarily hypopneas (71%), followed by obstructive (23%), central (4%), and mixed apneas (2%). As severity increased, the relative contribution of hypopneas and central apneas decreased, while that of obstructive apneas increased. The prevalence of Predominant CSA and Any CSA were 4.3% (26/606) and 8.4% (51/606) respectively. Being male, on opiates and having a high tetraplegic spinal cord injury were associated with CSA. SRH was identified in 26% (26/113) of the clinical sub-cohort. This is the largest study to characterize SDB in tetraplegia. It provides strong evidence that obstructive sleep apnea is the predominant SDB type; 9-18 times more prevalent than CSA. The prevalence of CSA was estimated to be 4%-8%, significantly lower than previously reported.


Protein and fatty acid composition of mesosomal vesicles and plasma membranes of Staphylococcus aureus.
Qualitatively, the protein and fatty acid composition of purified mesosomal vesicles and the plasma membrane isolated from Staphylococcus aureus ATCC 6538P are identical, the major difference between these two cellular components being only quantitative in nature. Mesosomal vesicles and plasma membranes, when subjected to acidic or neutral disk gel electrophoresis, exhibited more than 22 bands of protein. With urea-acetic acid gels, the plasma membrane had a higher concentration of "slower-migrating proteins" whereas "faster-migrating proteins" predominated in the mesosomal vesicles. With neutral disk gel electrophoresis, mesosomal vesicles exhibited one prominent protein band with an approximate molecular weight of 35,000 and which was four times greater than that found in the corresponding region on gels of the plasma membrane. Finally, fatty acid analyses by capillary column gas chromatography showed that although the fatty acid composition is the same, the fatty acid content in mesosomal vesicles is 48% greater than that of the plasma membrane. The dominant fatty acids in both of these cellular components are the iso and anteiso branched methyl C15, C17, and C19 fatty acids and comprise at least 85% of the total fatty acids extracted. These results show that distinct chemical differences exist between the mesosomal vesicles and the plasma membrane of Staphylococcus aureus.


Comprehensive analysis of the association between inflammation indexes and complications in patients undergoing pancreaticoduodenectomy.
During clinical practice, routine blood tests are commonly performed following pancreaticoduodenectomy (PD). However, the relationship between blood cell counts, inflammation-related indices, and postoperative complications remains unclear. We conducted a retrospective study, including patients who underwent PD from October 2018 to July 2023 at the First Hospital of Chongqing Medical University, and compared baseline characteristics and clinical outcomes among different groups. Neutrophil count (NC), platelet count (PLT), lymphocyte count (LC), systemic immune-inflammation index (SII), platelet-to-lymphocyte ratio (PLR), neutrophil-to-lymphocyte ratio (NLR), and the product of platelet count and neutrophil count (PPN) were derived from postoperative blood test results. We investigated the association between these indicators and outcomes using multivariable logistic regression and restricted cubic spline analysis. The predictive performance of these indicators was assessed by the area under the curve (AUC) of the receiver operating characteristic (ROC) curve and decision curve analysis (DCA). A total of 232 patients were included in this study. Multivariate logistic regression and restricted cubic spline analysis showed that all indicators, except for PLT, were associated with clinical postoperative pancreatic fistula (POPF). SII, NLR, and NC were linked to surgical site infection (SSI), while SII, NLR, and PLR were correlated with CD3 complication. PLT levels were related to postoperative hemorrhage. SII (AUC: 0.729), NLR (AUC: 0.713), and NC (AUC: 0.706) effectively predicted clinical POPF. In patients undergoing PD, postoperative inflammation-related indices and blood cell counts are associated with various complications. NLR and PLT can serve as primary indicators post-surgery for monitoring complications.


The Efficacy of Administering Fruit-Derived Polyphenols to Improve Health Biomarkers, Exercise Performance and Related Physiological Responses.
Polyphenols are secondary metabolites involved in a myriad of critical processes in plants. Over recent decades, special attention has been paid to the anti-oxidative role of fruit-derived polyphenols in the human diet, with evidence supporting the contribution of polyphenols in the prevention of numerous non-communicable disease outcomes. However, due to the low concentration in biological fluids in vivo, the antioxidant properties of polyphenols seem to be related to an enhanced endogenous antioxidant capacity induced via signaling through the nuclear respiratory factor 2 pathway. Polyphenols also seem to possess anti-inflammatory and antioxidant properties and have been shown to enhance vascular function via nitric oxide mediated mechanisms. Consequently, there is rationale to support fruit-derived polyphenol supplementation to enhance exercise performance, possibly via improved muscle perfusion. Fruit-derived polyphenol supplementation in exercise studies have included a variety of fruits, e.g., New Zealand blackcurrant, pomegranate, and cherry, in the form of extracts (multicomponent or purified), juices and infusions to varying degrees of benefit. For example, research has yet to link the health-related benefits of black elderberry (Sambucus nigra L.) ingestion to exercise performance in spite of the purported health benefits associated with black elderberry provision in vitro and in vivo models, which has been attributed to their high antioxidant capacity and polyphenol content. This review summarizes the existing evidence supporting a beneficial effect of fruit-derived polyphenols on various biological processes and outlines the potential for black elderberry ingestion to improve nitric oxide production, exercise performance, and the associated physiological responses before-, during- and post-exercise.


Pectin-Zeolite-Based Wound Dressings with Controlled Albumin Release.
Hypoalbuminemia can lead to poor and delayed wound healing, while it is also associated with acute myocardial infarction, heart failure, malignancies, and COVID-19. In elective surgery, patients with low albumin have high risks of postoperative wound complications. Here, we propose a novel cost-effective wound dressing material based on low-methoxy pectin and NaA-zeolite particles with controlled albumin release properties. We focused on both albumin adsorption and release phenomena for wounds with excess exudate. Firstly, we investigated albumin dynamics and calculated electrostatic surfaces at experimental pH values in water by using molecular dynamics methods. Then, we studied in detail pectin-zeolite hydrogels with both adsorption and diffusion into membrane methods using different pH values and albumin concentrations. To understand if uploaded albumin molecules preserved their secondary conformation in different formulations, we monitored the effect of pH and albumin concentration on the conformational changes in albumin after it was released from the hydrogels by using CD-UV spectroscopy analyses. Our results indicate that at pH 6.4, BSA-containing films preserved the protein's folded structure while the protein was being released to the external buffer solutions. In vitro wound healing assay indicated that albumin-loaded hydrogels showed no toxic effects on the fibroblast cells.


Stable transduction of quiescent T cells without induction of cycle progression by a novel lentiviral vector pseudotyped with measles virus glycoproteins.
A major limitation of current lentiviral vectors (LVs) is their inability to govern efficient gene transfer into quiescent cells such as primary T cells, which hampers their application for gene therapy. Here we generated high-titer LVs incorporating Edmonston measles virus (MV) glycoproteins H and F on their surface. They allowed efficient transduction through the MV receptors, SLAM and CD46, both present on blood T cells. Indeed, these H/F-displaying vectors outperformed by far VSV-G-LVs for the transduction of IL-7-prestimulated T cells. More importantly, a single exposure to these H/F-LVs allowed efficient gene transfer in quiescent T cells, which are not permissive for VSV-G-LVs that need cell-cycle entry into the G1b phase for efficient transduction. High-level transduction of resting memory (50%) and naive (11%) T cells with H/F-LVs, which seemed to occur mainly through SLAM, was not at cost of cell-cycle entry or of target T-cell activation. Finally, the naive or memory phenotypes of transduced resting T cells were maintained and no changes in cytokine profiles were detected, suggesting that T-cell populations were not skewed. Thus, H/F-LV transduction of resting T cells overcomes the limitation of current lentiviral vectors and may improve the efficacy of T cell-based gene therapy.


Treatment approach and survival from glioblastoma: results from a population-based retrospective cohort study from Western Norway.
To evaluate treatment and survival from glioblastoma in a real-world setting. A population-based retrospective cohort study from Western Norway. 363 patients aged 18 years or older diagnosed with glioblastoma between 1 January 2007 and 31 December 2014. Overall survival and survival rates determined by Kaplan-Meier method, groups compared by log-rank test. Associations between clinical characteristics and treatment approach assessed by logistic regression. Associations between treatment approach and outcome analysed by Cox regression. Median overall survival was 10.2 months (95% CI 9.1 to 11.3). Resection was performed in 221 patients (60.9%), and was inversely associated with age over 70 years, higher comorbidity burden, deep-seated tumour localisation and multifocality. Median survival was 13.7 months (95% CI 12.1 to 15.4) in patients undergoing tumour resection, 8.3 months (95% CI 6.6 to 9.9) in patients undergoing biopsy and 4.5 months (95% CI 4.0 to 5.1) in patients where no surgical intervention was performed. Chemoradiotherapy according to the Stupp protocol was given to 157 patients (43%). Age over 70 years, higher comorbidity burden and cognitive impairment were associated with less intensive chemoradiotherapy. Median survival was 16.3 months (95% CI 14.1 to 18.5), 7.9 months (95% CI 6.7 to 9.0) and 2.0 months (95% CI 0.9 to 3.2) in patients treated according to the Stupp protocol, with less intensive chemoradiotherapy and with best supportive care, respectively. Surgical resection (HR 0.61 (95% CI 0.47 to 0.79)) and chemoradiotherapy according to the Stupp protocol (HR 0.09 (95% CI 0.06 to 0.15)) were strongly associated with favourable overall survival, when adjusted for clinical variables. In a real-world setting, less than half of the patients received full-course chemoradiotherapy, with a median survival comparable to results from clinical trials. Survival was considerably worse in patients receiving less intensive treatment. Our results point out a substantial risk of undertreating glioblastoma, especially in elderly patients.


Suppression of Escherichia coli O157:H7 by Dung Beetles (Coleoptera: Scarabaeidae) using the lowbush blueberry agroecosystem as a model system.
Wildlife as a source of microbial contamination is a food safety concern. Deer feces (scat) have been determined as a point source for Escherichia coli O157:H7 contamination of fresh produce. The ecological role of the scooped scarab (Onthophagus hecate (Panzer)), a generalist dung beetle species common in Maine blueberry fields, was explored as a biological control agent and alternatively as a pathogen vector between deer scat and food. A large-scale field survey of wildlife scat indicated that pathogenic E. coli O157:H7 was present, albeit at a low prevalence (1.9% of samples, n = 318), in the Maine lowbush blueberry agroecosystem. A manipulative field experiment verified that, should contact occur between deer scat and blueberry plants and fruit during the summer, contamination with E. coli O157:H7 can occur and persist for more than 72 h. For both the positive control and an experimental scat inoculation treatment, the levels of the bacterial population decreased over time, but at different rates (treatment x time interaction: F (1.9,18.8) = 358.486, P < 0.0001). The positive control inoculation, which resulted in a higher initial E. coli level on fruit, decayed at a faster rate than inoculation of fruit via scat in the experimental treatment. We conducted 2 laboratory studies to elucidate aspects of dung beetle feeding ecology as it relates to suppression of E. coli O157:H7 from deer scat to lowbush blueberry fruit. In both experiments, dung beetles buried the same amount of scat whether or not the scat was inoculated with the pathogen (F(1,6) = 0.001; P = 0.999 and (F (2,17) = 4.10, P = 0.147). Beetles feeding on E. coli inoculated deer scat were not found to vector the pathogen to fruit. In two studies, beetles lowered the amount of pathogenic E. coli persisting in soils compared to soils without beetles (F (2,9) = 7.757; P = 0.05 and F (2,17) = 8.0621, P = 0.004). Our study suggests that the dung beetle species, Onthophagus hecate, has the potential to contribute to the suppression of E. coli O157:H7 in agricultural landscapes.


Incorporating β-cyclodextrin with ZnO nanorods: a potentiometric strategy for selectivity and detection of dopamine.
We describe a chemical sensor based on a simple synthesis of zinc oxide nanorods (ZNRs) for the detection of dopamine molecules by a potentiometric approach. The polar nature of dopamine leads to a change of surface charges on the ZNR surface via metal ligand bond formation which results in a measurable electrical signal. ZNRs were grown on a gold-coated glass substrate by a low temperature aqueous chemical growth (ACG) method. Polymeric membranes incorporating β-cyclodextrin (β-CD) and potassium tetrakis (4-chlorophenyl) borate was immobilized on the ZNR surface. The fabricated electrodes were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM) techniques. The grown ZNRs were well aligned and exhibited good crystal quality. The present sensor system displays a stable potential response for the detection of dopamine in 10(-2) mol·L(-1) acetic acid/sodium acetate buffer solution at pH 5.45 within a wide concentration range of 1 × 10(-6) M(-1) × 10(-1) M, with sensitivity of 49 mV/decade. The electrode shows a good response time (less than 10 s) and excellent repeatability. This finding can contribute to routine analysis in laboratories studying the neuropharmacology of catecholamines. Moreover, the metal-ligand bonds can be further exploited to detect DA receptors, and for bio-imaging applications.


The role of Pannexin-1 channels, ATP, and purinergic receptors in the pathogenesis of HIV and SARS-CoV-2.
Infectious agents such as human immune deficiency virus-1 (HIV) and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) use host proteins to infect, replicate, and induce inflammation within the host. A critical component of these diseases is the axis between pannexin-1 channels, extracellular ATP, and purinergic receptors. Here, we describe the potential therapeutic role of Pannexin-1/purinergic approaches to prevent or reduce the devastating consequences of these pathogens.


Containment, Health, and Social Policies in the Time of COVID-19 - Determinants and Outcomes of Initial Responses across 120 Countries.
The coronavirus disease 2019 (COVID-19) pandemic has triggered an unprecedented number of policy responses around the world across multiple policy domains. While governments have combined containment and health policies with social policies (CHSPs) during the initial phase of the pandemic in various ways, the current literature offers little knowledge of the patterns of these combinations and their determinants and outcomes. This paper fills this gap by investigating CHSP combinations across ≥120 countries. We further examined whether the CHSP response was determined by political regimes or compensation hypotheses-serving the purposes of responding to pre-existing economic downturns, inequality or social unrest. We also investigated the associations between CHSP responses and mobility, virus infection and unemployment. Using policy data from the Oxford COVID-19 Government Response Tracker, results from sequence analysis indicated that governments' CHSP responses could be clustered into five categories: high social policies (SPs), middle SPs, containment and health (CH) leading SPs, low SPs and gradual high SPs. We used multinomial regression models to investigate determinants of CHSP responses. We found that CHSP responses did not differ by political regimes, and CHSP combinations were not driven by compensation hypotheses. Instead, gross domestic product per capita and government effectiveness were the key drivers for high levels of policy responses. We also found that low SP responses were associated with fewer mobility changes. Taken together, our findings suggest that lower-income countries required more support and resources in order for them to adopt necessary CH and SP responses.


Osteoblast response to dimethacrylate composites varying in composition, conversion and roughness using a combinatorial approach.
Dimethacrylate polymers and composites are seeing increased usage in orthopedics. As these applications require the material to integrate with the surrounding tissues, direct contact cytotoxicity assays should be used to assess the biocompatibility. This study utilized a combinatorial testing platform to evaluate the cell response to dimethacrylate composites with a variety of properties on a single sample. MC3T3-E1 pre-osteoblasts were cultured directly on composites with varying filler content, filler type, degree of conversion (DC), and surface topography. Cell viability, density, and area depended on an interplay of the material properties, with low DC causing a reduction in cell area but having minimal effect on cell viability, high filler content causing an increase in cell density, and filler content/type altering the surface roughness as a function of DC. The combinatorial testing platform successfully quantified the effects of numerous material properties on several aspects of the osteoblast response.


Ciliation and gene expression distinguish between node and posterior notochord in the mammalian embryo.
The mammalian node, the functional equivalent of the frog dorsal blastoporal lip (Spemann's organizer), was originally described by Viktor Hensen in 1876 in the rabbit embryo as a mass of cells at the anterior end of the primitive streak. Today, the term "node" is commonly used to describe a bilaminar epithelial groove presenting itself as an indentation or "pit" at the distal tip of the mouse egg cylinder, and cilia on its ventral side are held responsible for molecular laterality (left-right) determination. We find that Hensen's node in the rabbit is devoid of cilia, and that ciliated cells are restricted to the notochordal plate, which emerges from the node rostrally. In a comparative approach, we use the organizer marker gene Goosecoid (Gsc) to show that a region of densely packed epithelium-like cells at the anterior end of the primitive streak represents the node in mouse and rabbit and is covered ventrally by a hypoblast (termed "visceral endoderm" in the mouse). Expression of Nodal, a gene intricately involved in the determination of vertebrate laterality, delineates the wide plate-like posterior segment of the notochord in the rabbit and mouse, which in the latter is represented by the indentation frequently termed "the node." Similarly characteristic ciliation and nodal expression exists in Xenopus neurula embryos in the gastrocoel roof plate (GRP), i.e., at the posterior end of the notochord anterior to the blastoporal lip. Our data suggest that (1) a posterior segment of the notochord, here termed PNC (for posterior notochord), is characterized by features known to be involved in laterality determination, (2) the GRP in Xenopus is equivalent to the mammalian PNC, and (3) the mammalian node as defined by organizer gene expression is devoid of cilia and most likely not directly involved in laterality determination.


Structure of the O-antigen of Salmonella O66 and the genetic basis for similarity and differences between the closely related O-antigens of Escherichia coli O166 and Salmonella O66.
O-antigen is a component of the outer membrane of Gram-negative bacteria and is one of the most variable cell surface constituents, leading to major antigenic variability. The O-antigen forms the basis for bacterial serotyping. In this study, the O-antigen structure of Salmonella O66 was established, which differs from the known O-antigen structure of Escherichia coli O166 only in one linkage (most likely the linkage between the O-units) and O-acetylation. The O-antigen gene clusters of Salmonella O66 and E. coli O166 were found to have similar organizations, the only exception being that in Salmonella O66, the wzy gene is replaced by a non-coding region. The function of the wzy gene in E. coli O166 was confirmed by the construction and analysis of deletion and trans-complementation mutants. It is proposed that a functional wzy gene located outside the O-antigen gene cluster is involved in Salmonella O66 O-antigen biosynthesis, as has been reported previously in Salmonella serogroups A, B and D1. The sequence identity for the corresponding genes between the O-antigen gene clusters of Salmonella O66 and E. coli O166 ranges from 64 to 70 %, indicating that they may originate from a common ancestor. It is likely that after the species divergence, Salmonella O66 got its specific O-antigen form by inactivation of the wzy gene located in the O-antigen gene cluster and acquisition of two new genes (a wzy gene and a prophage gene for O-acetyl modification) both residing outside the O-antigen gene cluster.


AB5075, a Highly Virulent Isolate of Acinetobacter baumannii, as a Model Strain for the Evaluation of Pathogenesis and Antimicrobial Treatments.
Acinetobacter baumannii is recognized as an emerging bacterial pathogen because of traits such as prolonged survival in a desiccated state, effective nosocomial transmission, and an inherent ability to acquire antibiotic resistance genes. A pressing need in the field of A. baumannii research is a suitable model strain that is representative of current clinical isolates, is highly virulent in established animal models, and can be genetically manipulated. To identify a suitable strain, a genetically diverse set of recent U.S. military clinical isolates was assessed. Pulsed-field gel electrophoresis and multiplex PCR determined the genetic diversity of 33 A. baumannii isolates. Subsequently, five representative isolates were tested in murine pulmonary and Galleria mellonella models of infection. Infections with one strain, AB5075, were considerably more severe in both animal models than those with other isolates, as there was a significant decrease in survival rates. AB5075 also caused osteomyelitis in a rat open fracture model, while another isolate did not. Additionally, a Tn5 transposon library was successfully generated in AB5075, and the insertion of exogenous genes into the AB5075 chromosome via Tn7 was completed, suggesting that this isolate may be genetically amenable for research purposes. Finally, proof-of-concept experiments with the antibiotic rifampin showed that this strain can be used in animal models to assess therapies under numerous parameters, including survival rates and lung bacterial burden. We propose that AB5075 can serve as a model strain for A. baumannii pathogenesis due to its relatively recent isolation, multidrug resistance, reproducible virulence in animal models, and genetic tractability. The incidence of A. baumannii infections has increased over the last decade, and unfortunately, so has antibiotic resistance in this bacterial species. A. baumannii is now responsible for more than 10% of all hospital-acquired infections in the United States and has a >50% mortality rate in patients with sepsis and pneumonia. Most research on the pathogenicity of A. baumannii focused on isolates that are not truly representative of current multidrug-resistant strains isolated from patients. After screening of a panel of isolates in different in vitro and in vivo assays, the strain AB5075 was selected as more suitable for research because of its antibiotic resistance profile and increased virulence in animal models. Moreover, AB5075 is susceptible to tetracycline and hygromycin, which makes it amenable to genetic manipulation. Taken together, these traits make AB5075 a good candidate for use in studying virulence and pathogenicity of this species and testing novel antimicrobials.


Guided plasmonic modes in nanorod assemblies: strong electromagnetic coupling regime.
We demonstrate that the coupling between plasmonic modes of oriented metallic nanorods results in the formation of an extended (guided) plasmonic mode of the nanorod array. The electromagnetic field distribution associated to this mode is found to be concentrated between the nanorods within the assembly and propagates normally to the nanorod long axes, similar to a photonic mode waveguided by an anisotropic slab. This collective plasmonic mode determines the optical properties of nanorod assemblies and can be tuned in a wide spectral range by changing the nanorod array geometry. This geometry represents a unique opportunity for light guiding applications and manipulation at the nanoscale as well as sensing applications and development of molecular plasmonic devices.