Caging Cationic Polymer Brush-Coated Plasmonic Nanostructures for Traceable Selective Antimicrobial Activities.
Cationic polymers are under intense research to achieve prominent antimicrobial activity. However, the cellular and in vivo toxicity caused by nonspecific electrostatic interaction has become a major challenge for their practical applications. Here, the development of a "caging" strategy based on the use of a block copolymer consisting of a stealth block and an anionic block that undergoes degradation in presence of enzymes secreted by selective bacterial pathogens of interest is reported. The results have shown that antimicrobial cationic polymer brushes-coated gold nanorods (AuNRs) can be caged by the block polymer of poly(ethylene glycol) and anionic, lipase-degradable block of ε-caprolactone and methacrylic acid copolymer to afford neutrally charged surfaces. The caged AuNRs are activated by lipase released by bacteria of interest to endow an excellent bactericidal effect but show minimal binding and toxicity against mammalian cells and nonspecific bacteria that do not produce lipase. In this design, AuNRs play multifunctional roles as the scaffolds for polymer brushes, photothermal transducers, and imaging probes for traceable delivery of the activation and delivery of bactericidal cationic polymer brushes. The caging strategy opens new opportunities for the safe delivery of antimicrobial materials for the treatment of bacterial infections.


Anti-B-50 (GAP-43) antibodies decrease exocytosis of glutamate in permeated synaptosomes.
The involvement of the protein kinase C substrate, B-50 (GAP-43), in the release of glutamate from small clear-cored vesicles in streptolysin-O-permeated synaptosomes was studied by using anti-B-50 antibodies. Glutamate release was induced from endogenous as well as 3H-labelled pools in a [Ca(2+)]-dependent manner. This Ca(2+)-induced release was partially ATP dependent and blocked by the light-chain fragment of tetanus toxin, demonstrating its vesicular nature. Comparison of the effects of anti-B-50 antibodies on glutamate and noradrenaline release from permeated synaptosomes revealed two major differences. Firstly, Ca(2+)-induced glutamate release was decreased only partially by anti-B-50 antibodies, whereas Ca(2+)-induced noradrenaline release was inhibited almost completely. Secondly, anti-B-50 antibodies significantly reduced basal glutamate release, but did not affect basal noradrenaline release. In view of the differences in exocytotic mechanisms of small clear-cored vesicles and large dense-cored vesicles, these data indicate that B-50 is important in the regulation of exocytosis of both types of neurotransmitters, probably at stages of vesicle recycling and/or vesicle recruitment, rather than in the Ca(2+)-induced fusion step.


Salt effects on the conformational stability of the visual G-protein-coupled receptor rhodopsin.
Membrane protein stability is a key parameter with important physiological and practical implications. Inorganic salts affect protein stability, but the mechanisms of their interactions with membrane proteins are not completely understood. We have undertaken the study of a prototypical G-protein-coupled receptor, the α-helical membrane protein rhodopsin from vertebrate retina, and explored the effects of inorganic salts on the thermal decay properties of both its inactive and photoactivated states. Under high salt concentrations, rhodopsin significantly increased its activation enthalpy change for thermal bleaching, whereas acid denaturation affected the formation of a denatured loose-bundle state for both the active and inactive conformations. This behavior seems to correlate with changes in protonated Schiff-base hydrolysis. However, chromophore regeneration with the 11-cis-retinal chromophore and MetarhodopsinII decay kinetics were slower only in the presence of sodium chloride, suggesting that in this case, the underlying phenomenon may be linked to the activation of rhodopsin and the retinal release processes. Furthermore, the melting temperature, determined by means of circular dichroism and differential scanning calorimetry measurements, was increased in the presence of high salt concentrations. The observed effects on rhodopsin could indicate that salts favor electrostatic interactions in the retinal binding pocket and indirectly favor hydrophobic interactions at the membrane protein receptor core. These effects can be exploited in applications where the stability of membrane proteins in solution is highly desirable.


Extracellular vesicle-embedded materials.
Extracellular vesicles (EVs) are small membrane-bound vesicles released by cells. EVs are emerging as a promising class of therapeutic entity that could be adapted in formulation due to their lack of immunogenicity and targeting capabilities. EVs have been shown to have similar regenerative and therapeutic effects to their parental cells and also have potential in disease diagnosis. To improve the therapeutic potential of EVs, researchers have developed various strategies for modifying them, including genetic engineering and chemical modifications which have been examined to confer target specificity and prevent rapid clearance after systematic injection. Formulation efforts have focused on utilising hydrogel and nano-formulation strategies to increase the persistence of EV localisation in a specific tissue or organ. Researchers have also used biomaterials or bioscaffolds to deliver EVs directly to disease sites and prolong EV release and exposure. This review provides an in-depth examination of the material design of EV delivery systems, highlighting the impact of the material properties on the molecular interactions and the maintenance of EV stability and function. The various characteristics of materials designed to regulate the stability, release rate and biodistribution of EVs are described. Other aspects of material design, including modification methods to improve the targeting of EVs, are also discussed. This review aims to offer an understanding of the strategies for designing EV delivery systems, and how they can be formulated to make the transition from laboratory research to clinical use.


Heterogeneity of astrocytes: Electrophysiological properties of juxtavascular astrocytes before and after brain injury.
Astrocyte heterogeneity is increasingly recognized, but still little is known about juxtavascular astrocytes with their somata directly adjacent to blood vessels, despite their importance after brain injury. As juxtavascular astrocytes originate from common progenitor cells, that is, have a clonal origin, they may intrinsically differ from other, non-juxtavascular astrocytes. To explore this, we examined the electrophysiological properties of these groups of astrocytes and the underlying ion channels. Using brain slices of BAC Aldh1l1-eGFP transgenic mice with astrocytes labeled by GFP expression, we compared juxtavascular and non-juxtavascular astrocytes in the somatosensory cortex by means of whole-cell patch-clamp recordings and immunohistochemical staining. Prior to injury, juxta- and non-juxtavascular astrocytes exhibit comparable electrophysiological properties with characteristic mostly passive conductance and a typical negative resting membrane potential. Immunohistochemical analysis of K+ channels showed that all astrocytes were Kir 4.1+ , but revealed an intriguing difference for Kv 4.3. The expression of Kv 4.3 in sibling astrocytes (non-juxtavascular, juxtavascular and pial) was dependent on their ontogenetic origin with lowest levels in juxtavascular astrocytes located in upper cortical layers. After traumatic brain injury (TBI), we found profound changes in the electrophysiological type of astrocytes with a predominance of non-passive properties and this pattern was significantly enriched in juxtavascular astrocytes. This was accompanied by pronounced down-regulation of Kir 4.1 in proliferating astrocytes, which was significantly more in juxtavascular compared to non-juxtavascular astrocytes. Taken together, TBI induces profound differences in electrophysiological properties between juxtavascular and non-juxtavascular astrocytes that might be related to the preponderance of juxtavascular astrocyte proliferation.


3-O-Laurylglyceryl ascorbate reinforces skin barrier function through not only the reduction of oxidative stress but also the activation of ceramide synthesis.
A higher trans-epidermal water loss (TEWL) occurs in rough skin, in elder skin and also in atopic dermatitis. An impaired skin barrier function is considered to be caused by an incomplete construction of the intercellular lamellar structure due to the quantitative reduction of ceramides. Since these symptoms coexist with oxidative stress, we hypothesized that impairment of the skin barrier function is accelerated by oxidative stress. Thus, the purpose of this study was to clarify the effect of oxidative stress on ceramide synthesis and to characterize whether antioxidants can improve skin barrier function. 3-O-Laurylglyceryl ascorbate (VC-3LG), which is a newly amphipathic derivative of ascorbic acid, was evaluated as a candidate antioxidant. We characterized the mRNA expression levels of serine palmitoyltransferase (SPT) in normal human epidermal keratinocytes (NHEKs) treated with H2 O2 using real-time PCR analysis. In order to evaluate the effect of VC-3LG on skin barrier function, we used several assays with reconstructed human epidermis equivalents (RHEEs). Ceramide synthesis was down-regulated in NHEKs by oxidative stress. Treatment with VC-3LG abrogated the down-regulation of SPT mRNA in NHEKs caused by oxidative stress, and stimulated SPT mRNA expression levels. In experiments characterizing the antioxidative properties of VC-3LG, VC-3LG reduced oxidative stress in NHEKs by up-regulating catalase mRNA expression. In addition, VC-3LG stimulated the skin barrier function in RHEEs, which had lower TEWL values compared with untreated RHEEs. Furthermore, VC-3LG increased the quantity of ceramide in RHEEs. Taken together, we conclude that VC-3LG reinforces the skin barrier function due to its reduction of oxidative stress and its promotion of ceramide synthesis.


Inhibition of β-amyloid peptide self-assembly and cytotoxicity by poly(LVFF-co-β-amino ester).
The β-amyloid (Aβ) peptide plays an important role in the onset and progress of Alzheimer's disease (AD). Therefore, studies on inhibiting fibril formation and thus neurotoxicity of Aβ could be beneficial for the prevention and treatment of AD. We report a convenient synthetic approach for one-pot preparation of poly-(β-amino ester) copolymerized with the GGLVFF peptide, which is based on the frequently used inhibitor LVFF. The copolymer was found to efficiently inhibit the aggregation and fibrillation of Aβ42 by using fluorescence assay and atomic force microscopy. Reduced β-sheet formation of Aβ42 peptide after addition of copolymer was observed by circular dichroism. Furthermore, the cell viability assay confirmed that the toxicity of Aβ42 for SH-SY5Y cells was markedly reduced in the presence of copolymer. This could be beneficial for AD prevention and treatment and also for the molecular design of inhibitory agents for other amyloidoses.


Cost-utility analysis of total knee arthroplasty using 10-year data from a randomised controlled trial: Implant design influences quality-adjusted life year gain.
The aim of this study was to perform a cost-utility analysis for total knee arthroplasty (TKA) over a 10-year follow up period. Two-hundred and twelve patients were randomised to receive either a Kinemax or a Triathlon TKA. Patients were assessed pre-operatively, and at 6 months, and 1, 3, 7 and 10 years postoperatively. The costs of the primary and revision surgery were accounted for. One-year quality-adjusted life year (QALY) gain was used to estimate 10-year gains using the established annual health gain discounts. Forty-eight patients died and eight were revised during the follow up period. Overall QALY gain per patient over the 10-year period was 2.594 and the cost per patient was £6559, which resulted in a cost per QALY of £2761 at 10 years. The Triathlon group had a significantly greater QALY gain compared with the Kinemax (mean difference (MD) 0.53, 95% CI 0.03-1.03, P = 0.02), which resulted in a cost per QALY for the Triathlon group of £2521 compared with £3107 for the Kinemax group at 10 years. The 5% annual discount resulted in a significantly lower QALY gain (MD 0.135, 95% CI 0.201-0.354, P = 0.002), whereas the 3.5% annual discount resulted in non-significant difference in QALY gain compared with the actual gain (MD 0.021, 95% CI -0.084 to 0.077, P = 0.292). TKA was a cost-effective intervention, and the Triathlon was associated with a greater cost effectiveness at 10 years. The 3.5% annual discounts for QALY gain would seem to be the most accurate, with an underestimation being observed with the 5% discount.


Hepatic gluconeogenesis and Krebs cycle fluxes in a CCl4 model of acute liver failure.
Acute liver failure was induced in rats by CCl4 administration and its effects on the hepatic Krebs cycle and gluconeogenic fluxes were evaluated in situ by 13C NMR isotopomer analysis of hepatic glucose following infusion of [U-13C]propionate. In fed animals, CCl4 injury caused a significant increase in relative gluconeogenic flux from 0.80+/-0.10 to 1.34 +/-0.24 times the flux through citrate synthase (p<0.01). In 24-h fasted animals, CCl4-injury also significantly increased relative gluconeogenic flux from 1.36+/-0.16 to 1.80+/-0.22 times the flux through citrate synthase (p<0.01). Recycling of PEP via pyruvate and oxaloacetate was extensive under all conditions and was not significantly altered by CCl4 injury. CCl4 injury significantly reduced hepatic glucose output by 26% (42.8+/-7.3 vs 58.1+/-2.4 micromol/kg/min, p=0.005), which was attributed to a 26% decrease in absolute gluconeogenic flux from PEP (85.6+/-14.6 vs 116+/-4.8 micromol/kg/min, p<0.01). These changes were accompanied by a 47% reduction in absolute citrate synthase flux (90.6+/-8.0 to 47.6+/-8.0 micromol/kg/min, p<0.005), indicating that oxidative Krebs cycle flux was more susceptible to CCl4 injury. The reduction in absolute fluxes indicate a significant loss of hepatic metabolic capacity, while the significant increases in relative gluconeogenic fluxes suggest a reorganization of metabolic activity towards preserving hepatic glucose output.


Patient-specific modeling of cardiovascular and respiratory dynamics during hypercapnia.
This study develops a lumped cardiovascular-respiratory system-level model that incorporates patient-specific data to predict cardiorespiratory response to hypercapnia (increased CO(2) partial pressure) for a patient with congestive heart failure (CHF). In particular, the study focuses on predicting cerebral CO(2) reactivity, which can be defined as the ability of vessels in the cerebral vasculature to expand or contract in response CO(2) induced challenges. It is difficult to characterize cerebral CO(2) reactivity directly from measurements, since no methods exist to dynamically measure vasomotion of vessels in the cerebral vasculature. In this study we show how mathematical modeling can be combined with available data to predict cerebral CO(2) reactivity via dynamic predictions of cerebral vascular resistance, which can be directly related to vasomotion of vessels in the cerebral vasculature. To this end we have developed a coupled cardiovascular and respiratory model that predicts blood pressure, flow, and concentration of gasses (CO(2) and O(2)) in the systemic, cerebral, and pulmonary arteries and veins. Cerebral vascular resistance is incorporated via a model parameter separating cerebral arteries and veins. The model was adapted to a specific patient using parameter estimation combined with sensitivity analysis and subset selection. These techniques allowed estimation of cerebral vascular resistance along with other cardiovascular and respiratory parameters. Parameter estimation was carried out during eucapnia (breathing room air), first for the cardiovascular model and then for the respiratory model. Then, hypercapnia was introduced by increasing inspired CO(2) partial pressure. During eucapnia, seven cardiovascular parameters and four respiratory parameters was be identified and estimated, including cerebral and systemic resistance. During the transition from eucapnia to hypercapnia, the model predicted a drop in cerebral vascular resistance consistent with cerebral vasodilation.


Phase angle is related to physical function and quality of life in preoperative patients with lumbar spinal stenosis.
Lumbar spinal stenosis (LSS) can interfere with daily life and quality of life (QOL). Evaluating physical function and QOL and helping patients to improve is the focus of rehabilitation. Phase angle (PhA) assessment is widely used to measure body composition and is considered an indicator of physical function and QOL. This study investigated the relationship between PhA and physical function, physical activity, and QOL in patients with LSS. PhA, handgrip strength, walking speed, Timed Up and Go test (TUG), Life Space Assessment (LSA), Prognostic Nutritional Index (PNI), Japanese Orthopaedic Association Back Pain Evaluation Questionnaire (JOABPEQ), and EQ-5D were assessed and statistically analyzed. The study included 133 patients with LSS. Multiple regression analysis of PhA adjusted for age, sex, and body mass index (Model 1) and for Model 1 + PNI (Model 2) showed significant correlations (P < 0.05) with handgrip strength, walking speed, TUG, and LSA. Regarding QOL, PhA was significantly correlated (P < 0.05) with lumbar function in JOABPEQ. PhA was associated with physical function and QOL in patients with LSS and might be a new clinical indicator in this population.


Electrocardiographic changes in thyrotoxic periodic paralysis.
The electrocardiograms (ECGs) of 30 patients with hypokalaemic thyrotoxic periodic paralysis during and after paralysis were studied. During paralysis, typical features of hypokalaemia were seen in all patients with serum potassium levels of 2.8 mmol/l or less; above this level, the ECGs varied from non-diagnostic to those showing typical features of hypokalaemia. It was not possible to accurately predict the serum potassium level from the ECG except when either sinus arrest or heart block was present. Although extrasystoles have been reported to be common in hypokalaemia, none of the patients in this study had extrasystoles. Sinus arrest occurred in two patients and second degree atrio-ventricular block occurred in three patients, a finding which has not been reported in hypokalaemia.


Prealbumin improves death risk prediction of BNP-added Seattle Heart Failure Model: results from a pilot study in elderly chronic heart failure patients.
An accurate prognosis prediction represents a key element in chronic heart failure (CHF) management. Seattle Heart Failure Model (SHFM) prognostic power, a validated risk score for predicting mortality in CHF, is improved by adding B-type natriuretic peptide (BNP). We evaluated in a prospective study the incremental value of several biomarkers, linked to different biological domains, on death risk prediction of BNP-added SHFM. Troponin I (cTnI), norepinephrine, plasma renin activity, aldosterone, high sensitivity-C reactive protein (hs-CRP), tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), interleukin 2 soluble receptor, leptin, prealbumin, free malondialdehyde, and 15-F2t-isoprostane were measured in plasma from 142 consecutive ambulatory, non-diabetic stable CHF (mean NYHA-class 2.6) patients (mean age 75±8years). Calibration, discrimination, and risk reclassification of BNP-added SHFM were evaluated after individual biomarker addition. Individual addition of biomarkers to BNP-added SHFM did not improve death prediction, except for prealbumin (HR 0.49 CI: (0.31-0.76) p=0.002) and cTnI (HR 2.03 CI: (1.20-3.45) p=0.009). In fact, with respect to BNP-added SHFM (Harrell's C-statistic 0.702), prealbumin emerged as a stronger predictor of death showing the highest improvement in model discrimination (+0.021, p=0.033) and only a trend was observed for cTn I (+0.023, p=0.063). These biomarkers showed also the best reclassification statistic (Integrated Discrimination Improvement-IDI) at 1-year (IDI: cTnI, p=0.002; prealbumin, p=0.020), 2-years (IDI: cTnI, p=0.018; prealbumin: p=0.006) and 3-years of follow-up (IDI: cTnI p=0.024; prealbumin: p=0.012). Individual addition of prealbumin allows a more accurate prediction of mortality of BNP enriched SHFM in ambulatory elderly CHF suggesting its potential use in identifying those at high-risk that need nutritional surveillance.


Early rheumatoid-like synovial lesions in rabbits drinking cow's milk. I. Joint pathology.
Rheumatoid-like lesions developed in 9 of 25 (36%) Old English rabbits drinking cow's milk for 12 weeks. The incidence of lesions in male and female animals was similar. The majority of rabbits drinking cow's milk developed increased numbers of nucleated cells and raised percentages of T lymphocytes in their synovial fluids, compared to control rabbits. The cell counts and T cell percentages correlated with the severity of the histological lesions. No evidence of glomerulonephritis was observed in any of the experimental animals.


A response to readers' comments.
This is a response to readers' comments on our paper entitled "Critical role of NLRP3-caspase-1 pathway in age-dependent isoflurane-induced microglial inflammatory response and cognitive impairment" published in the Journal of Neuroinflammation this year.


Native valve infective endocarditis in a tertiary care center in a developing country (Tunisia).
The aim of the study was to describe the epidemiological and clinical aspects of native valve infective endocarditis (IE) in a Tunisian high-volume tertiary care center and to identify the predictors of outcome. Demographic, clinical, laboratory, and echocardiographic characteristics were examined in 134 patients who fulfilled the modified Duke criteria for native valve IE between January 1997 and December 2006. Logistic regression analysis was used to identify prognostic factors for death. Mean age was 34.22 years. Diagnosis was definite in 93% of cases. Median time to diagnosis was 21 days. Rheumatic heart disease (RHD) was the predominant (45%) underlying heart condition. One or more vegetations were detected in more than 93% of cases. The median size of vegetation was >15 mm in 28% of cases. In 66 cases (49%), cultures remained negative. Serology was positive in 15 cases, and in 4 cases leaflet culture identified the agent. The infective agent was identified in 87 cases (65%), causative microorganisms were mainly Staphylococci (n = 30, including 6 coagulase-negative Staphylococcus), and Streptococci (n = 32). Overall mortality was 19%. On multivariate analysis, congestive heart failure (hazard ratio = 5.34, 95% confidence interval 1.67 to 17.15, p = 0.005) and large vegetations (>15 mm; hazard ratio = 5.78, 95% confidence interval 1.84 to 18.32, p = 0.002) were predictive of in-hospital mortality but not neurological complications or staphylococcus IE. In conclusion, IE remains a serious disease affecting a young population in Tunisia, with RHD as still the most common underlying heart disease, and it is associated with a high mortality.


Small RNA transcriptomes of mangroves evolve adaptively in extreme environments.
MicroRNAs (miRNAs) and endogenous small interfering RNAs (siRNAs) are key players in plant stress responses. Here, we present the sRNA transcriptomes of mangroves Bruguiera gymnorrhiza and Kandelia candel. Comparative computational analyses and target predictions revealed that mangroves exhibit distinct sRNA regulatory networks that differ from those of glycophytes. A total of 32 known and three novel miRNA families were identified. Conserved and mangrove-specific miRNA targets were predicted; the latter were widely involved in stress responses. The known miRNAs showed differential expression between the mangroves and glycophytes, reminiscent of the adaptive stress-responsive changes in Arabidopsis. B. gymnorrhiza possessed highly abundant but less conserved TAS3 trans-acting siRNAs (tasiRNAs) in addition to tasiR-ARFs, with expanded potential targets. Our results indicate that the evolutionary alteration of sRNA expression levels and the rewiring of sRNA-regulatory networks are important mechanisms underlying stress adaptation. We also identified sRNAs that are involved in salt and/or drought tolerance and nutrient homeostasis as possible contributors to mangrove success in stressful environments.


Sex chromosome pairing patterns in male mice of novel Sxr genotypes.
The influence of the sex-reversal factor (Sxr) on X and Y chromosome pairing was examined by comparing males with "novel" and "standard" Sxr genotypes. The novel Sxr males were exceptional in carrying Sxr on their X rather than their Y chromosome, or homozygously on both their X and Y chromosomes, or on a Y chromosome of different origin to that on which the factor arose. Regardless of its chromosomal location, Sxr was found to elevate the frequency of X-Y separation. Univalent X and Y chromosomes were observed to undergo self-association in a variable proportion of spermatocytes of all Sxr-carrying males. There was a suggestion that chromosomal location of the factor could influence the frequency of univalent self-association. Our observations do not support the published hypothesis of Y self-pairing as the cause of the elevated rate of X-Y separation at pachytene in Sxr-carrying males. Rather, they suggest that heterozygosity due to the presence of Sxr in the XY pairing region may be sufficient to disrupt pairing and cause univalence, or alternatively, that Sxr is an inefficient promoter of X-Y pairing initiation.


Bioinformatics Tools for Bulk Gene Expression Deconvolution in Diabetic Retinopathy.
Retinal neovascularization is one of the leading causes of vision loss and a hallmark of proliferative diabetic retinopathy (PDR). The immune system is observed to be involved in the pathogenesis of diabetic retinopathy (DR). The specific immune cell type that contributes to retinal neovascularization can be identified via a bioinformatics analysis of RNA sequencing (RNA-seq) data, known as deconvolution analysis. Previous study has identified the infiltration of macrophages in the retina of rats with hypoxia-induced retinal neovascularization and patients with PDR through a deconvolution algorithm, known as CIBERSORTx. Here, we describe the protocols of using CIBERSORTx to perform the deconvolution analysis and downstream analysis of RNA-seq data.


Duodenal atresia--60 years of follow-up (case report).
The first surviving patient with duodenal atresia operated upon by Dr. Ernst Copenhagen has been followed up for 60 years and is perfectly well.


Abdominal compartment syndrome in traumatic hemorrhagic shock: is there a fluid resuscitation inflection point associated with increased risk?
The volume of fluid administered during trauma resuscitation correlates with the risk of abdominal compartment syndrome (ACS). The exact volume at which this risk rises is uncertain. We established the inflection point for ACS risk during shock resuscitation. Using the Glue Grant database, patients aged ≥16 years with ACS were compared with those without ACS (no-ACS). Stepwise analysis of the sum or difference of the mean total fluid volume (TV)/kg, TV and/or body weight, (μ) and standard deviations (σ) vs % ACS at each point was used to determine the fluid inflection point. A total of 1,976 patients were included, of which 122 (6.2%) had ACS. Compared with no-ACS, ACS patients had a higher emergency room lactate (5.8 ± 3.0 vs 4.5 ± 2.8, P < .001), international normalized ratio (1.8 ± 1.5 vs 1.4 ± .8, P < .001), and mortality (37.7% vs 14.6%, P < .001). ACS group received a higher TV/kg (498 ± 268 mL/kg vs 293 ± 171 mL/kg, P < .001) than no-ACS. The % ACS increased exponentially with the sum of μ and incremental σ, with the sharpest increase occurring at TV and/or body weight = μ + 3σ or 1,302 mL/kg. There is a dramatic rise in ACS risk after 1,302 mL/kg of fluid is administered. This plot could serve as a guide in limiting the ACS risk during resuscitation.


A data-driven respiratory motion estimation approach for PET based on time-of-flight weighted positron emission particle tracking.
Respiratory motion of patients during positron emission tomography (PET)/computed tomography (CT) imaging affects both image quality and quantitative accuracy. Hardware-based motion estimation, which is the current clinical standard, requires initial setup, maintenance, and calibration of the equipment, and can be associated with patient discomfort. Data-driven techniques are an active area of research with limited exploration into lesion-specific motion estimation. This paper introduces a time-of-flight (TOF)-weighted positron emission particle tracking (PEPT) algorithm that facilitates lesion-specific respiratory motion estimation from raw listmode PET data. The TOF-PEPT algorithm was implemented and investigated under different scenarios: (a) a phantom study with a point source and an Anzai band for respiratory motion tracking; (b) a phantom study with a point source only, no Anzai band; (c) two clinical studies with point sources and the Anzai band; (d) two clinical studies with point sources only, no Anzai band; and (e) two clinical studies using lesions/internal regions instead of point sources and no Anzai band. For studies with radioactive point sources, they were placed on patients during PET/CT imaging. The motion tracking was performed using a preselected region of interest (ROI), manually drawn around point sources or lesions on reconstructed images. The extracted motion signals were compared with the Anzai band when applicable. For the purposes of additional comparison, a center-of-mass (COM) algorithm was implemented both with and without the use of TOF information. Using the motion estimate from each method, amplitude-based gating was applied, and gated images were reconstructed. The TOF-PEPT algorithm is shown to successfully determine the respiratory motion for both phantom and clinical studies. The derived motion signals correlated well with the Anzai band; correlation coefficients of 0.99 and 0.94-0.97 were obtained for the phantom study and the clinical studies, respectively. TOF-PEPT was found to be 13-38% better correlated with the Anzai results than the COM methods. Maximum Standardized Uptake Values (SUVs) were used to quantitatively compare the reconstructed-gated images. In comparison with the ungated image, a 14-39% increase in the max SUV across several lesion areas and an 8.7% increase in the max SUV on the tracked lesion area were observed in the gated images based on TOF-PEPT. The distinct presence of lesions with reduced blurring effect and generally sharper images were readily apparent in all clinical studies. In addition, max SUVs were found to be 4-10% higher in the TOF-PEPT-based gated images than in those based on Anzai and COM methods. A PEPT- based algorithm has been presented for determining movement due to respiratory motion during PET/CT imaging. Gating based on the motion estimate is shown to quantifiably improve the image quality in both a controlled point source phantom study and in clinical data patient studies. The algorithm has the potential to facilitate true motion correction where the reconstruction algorithm can use all data available.


'Between the demands of truth and government': health practitioners, trust and immunisation work.
Applied and theoretical work on health governance and governmentality has often not engaged with the perspectives of practitioners and, in particular, with their understandings about trust. In this paper, we address these absences through looking at a critical technology of governmentality--child immunisation. We do this, first, by examining theoretical links between risk, trust and knowledge in relation to the governance of health. We then draw on findings from our secondary analysis of qualitative data based on interviews with key actors in primary care in Scotland charged with the delivery of a particular immunisation--the MMR vaccine. While many practitioners, like parents, have typically perceived immunisation--along with other public health initiatives-as obligatory and as part of 'good citizenship', we argue that in relation to MMR and in the context of clinical governance, practitioners are having to engage in complex negotiations about knowledge and trust--negotiations that are at the heart of governing health at a distance.


Thrombocyte functions, thrombelastograms and fibrinogen of healthy children in different age groups.
Platelet counts, adhesiveness, aggregation and spreading capacity, as well as the thrombopoiesis index, thrombelastogram and fibrinogen were examined in 74 healthy children aged from 2 to 14 years. Sex-linked differences were only found in the thrombopoiesis index. Age-linked differences showed in practically all the parameters, especially in the platelet counts, fibrinogen and thrombelastograms.


GlycA, a novel marker for low grade inflammation, reflects gut microbiome diversity and is more accurate than high sensitive CRP in reflecting metabolomic profile.
Gut microbiota is, along with adipose tissue, recognized as a source for many metabolic and inflammatory disturbances that may contribute to the individual's state of health. We investigated in cross-sectional setting the feasibility of utilizing GlycA, a novel low grade inflammatory marker, and traditional low grade inflammatory marker, high sensitivity CRP (hsCRP), in reflecting serum metabolomics status and gut microbiome diversity. Fasting serum samples of overweight/obese pregnant women (n = 335, gestational weeks: mean 13.8) were analysed for hsCRP by immunoassay, GlycA and metabolomics status by NMR metabolomics and faecal samples for gut microbiome diversity by metagenomics. The benefits of GlycA as a metabolic marker were investigated against hsCRP. The GlycA concentration correlated with more of the metabolomics markers (144 out of 157), than hsCRP (55 out of 157) (FDR < 0.05). The results remained essentially the same when potential confounding factors known to associate with GlycA and hsCRP levels were taken into account (P < 0.05). This was attributable to the detected correlations between GlycA and the constituents and concentrations of several sized VLDL-particles and branched chain amino acids, which were statistically non-significant with regard to hsCRP. GlycA, but not hsCRP, correlated inversely with gut microbiome diversity. GlycA is a superior marker than hsCRP in assessing the metabolomic profile and gut microbiome diversity. It is proposed that GlycA may act as a novel marker that reflects both the gut microbiome and adipose tissue originated metabolic aberrations; this proposal will need to be verified with regard to clinical outcomes. ClinicalTrials.gov, NCT01922791, August 14, 2013.


The detection of superoxide anion from the reaction of oxyhemoglobin and phenylhydrazine using EPR spectroscopy.
The low temperature EPR spectrum of a quickly reacted mixture of oxyhemoglobin and phenylhydrazine was studied. With the use of a computer, the spectral contribution of methemoglobin in the region of g = 2 was subtracted. The remaining spectrum was that of an axial free radical (g perpendicular = 2.00, g parallel = 2.06) having the magnetic parameters of superoxide anion. In the presence of superoxide dismutase, this axial radical was not seen, confirming that superoxide anion is indeed generated by the reaction.


Emission of volatile organic compounds from residential biomass burning and their rapid chemical transformations.
Biomass combustion releases a complex array of Volatile Organic Compounds (VOCs) that pose significant challenges to air quality and human health. Although biomass burning has been extensively studied at ecosystem levels, understanding the atmospheric transformation and impact on air quality of emissions in urban environments remains challenging due to complex sources and burning materials. In this study, we investigate the VOC emission rates and atmospheric chemical processing of predominantly wood burning emissions in a small urban centre in Greece. Ioannina is situated in a valley within the Dinaric Alps and experiences intense atmospheric pollution accumulation during winter due to its topography and high wood burning activity. During pollution event days, the ambient mixing ratios of key VOC species were found to be similar to those reported for major urban centres worldwide. Positive matrix factorisation (PMF) analysis revealed that biomass burning was the dominant emission source (>50 %), representing two thirds of OH reactivity, which indicates a highly reactive atmospheric mixture. Calculated OH reactivity ranges from 5 s-1 to an unprecedented 278 s-1, and averages at 93 ± 66 s-1 at 9 PM, indicating the presence of exceptionally reactive VOCs. The highly pronounced photochemical formation of organic acids coincided with the formation of ozone, highlighting the significance of secondary formation of pollutants in poorly ventilated urban areas. Our findings underscore the pressing need to transition from wood burning to environmentally friendly sources of energy in poorly ventilated urban areas, in order to improve air quality and safeguard public health.


Schistosoma japonicum: inhibition of Mago nashi gene expression by shRNA-mediated RNA interference.
RNA interference (RNAi) mediated by short interfering RNA (siRNA) is a powerful reverse genetics tool and holds enormous therapeutic potential for various diseases, including parasite infections. siRNAs bind their complementary mRNA and lead to degradation of their specific mRNA targets. RNAi has been widely used for functional analysis of specific genes in various cells and organisms. In this paper, we tested the potential of silencing the expression of the Mago nashi gene in Schistosoma japonicum by siRNAs derived from shRNA expressed by mammalian Pol III promoter H1. Schistosomula, transformed from cercariae by mechanical shearing of the tails, were electroporated with Mago nashi shRNA expression vector. Aliquots of parasites were harvested at days 1, 3, and 5 after electroporation, respectively. Levels of Mago nashi mRNA and protein were determined by RT-PCR and Western blotting analysis. The results showed that shRNA expressed from mammalian Pol III promoter H1 specifically reduced the levels of Mago nashi mRNA and proteins in S. japonicum. Changes in testicular lobes were apparent when parasites were introduced into mammalian hosts. Thus, vector-mediated gene silencing is applicable to S. japonicum, which provides a means for the functional analysis of genes in this organism.


[Effects of acupuncture on serum insulin antibody and tumor necrosis factor alpha in the experimental rat with insulin resistance].
To observe effects of acupuncture and diet on insulin resistance (IR) and to probe the mechanism. Forty SID rats were equally divided into 5 groups: blank group (group I), model group I (group II), model group II (group III), acupuncture group I (group IV) and acupuncture group II (group V). The groups II, III, N and V were fed with high-fat-sugar-salt forage to made IR model, then the groups I, III and V were fed with normal forage, and the groups II and IV with the high-fat-sugar-salt forage, and the acupuncture groups IV and V received acupuncture treatment. Two weeks later, the fasting blood glucose (FBG), plasma insulin (INS), insulin sensitivity index (ISI), INS antibody and tumor necrosis factor alpha (TNF-alpha) were detected. As compared with group I, FBG and INS increased, ISI decreased in the group II (all P < 0.01); as compared with the group II , FBG and INS decreased (all P < 0.01) and ISI increased (P < 0.05, P < 0.01) in the group [II, IV, V; no case with INS antibody (+) in all groups; TNF-alpha in the group II increased compared with that of the group I (P < 0.01), and TNF-alpha in the group III, IV, V decreased compared with that of group II (P < 0.01). Acupuncture exerts a reversal effect on insulin resistance, and diet can promotes this effect. The mechanism is carried out possibly through decreasing the secretion of TNF-alpha.


Decreased gray matter concentration in the insular, orbitofrontal, cingulate, and temporal cortices of cocaine patients.
Structural deficiencies within limbic and prefrontal regions may contribute to the characteristic drug-seeking and drug-taking behaviors that prevail in persons dependent on cocaine. To date, a focal structural analysis of the brains of cocaine patients has not been undertaken. We used voxel based morphometry in conjunction with statistical parametric mapping on the structural magnetic resonance images of cocaine-dependent (n = 13) and cocaine-naive individuals (n = 16) to assess differences between the two groups in gray and white matter concentration. We report a decrease in gray matter concentration in the ventromedial orbitofrontal, anterior cingulate, anteroventral insular, and superior temporal cortices of cocaine patients in comparison to controls (p <.01 corrected for multiple comparisons). The average percentage decrease in gray matter concentration within a region ranged from 5% to 11%. White matter concentration did not differ between groups. We conclude that the brains of cocaine patients are structurally dissimilar from those of nondrug-using controls. The differences were detected in regions involved in decision-making, behavioral inhibition and assignation of emotional valence to environmental stimuli and, hence, may contribute to some of the behavioral deficits characteristic of chronic cocaine users.


cDNA cloning and differential expression patterns of ascorbate peroxidase during post-harvest in Brassica rapa L.
Ascorbate is an antioxidant and a cofactor of many dioxygenases in plant and animal cell metabolism. A well-recognized enzyme consuming ascorbate is ascorbate peroxidase (APX), which catalyses the reduction of hydrogen peroxide to water with the simultaneous oxidation of ascorbate with a high specificity. The isolation and characterisation of new Apx cDNAs, could provide new insights about the physiological roles and regulation of these enzymes. In this work chloroplastic (Br-chlApx) and cytosolic (Br-cApx) isoform transcripts were isolated by RT-PCR in Brassica rapa and expression changes were analysed by semi-quantitative RT-PCR performed in different tissues (layer, stalk and florets) at different days (0, 4 and 14 day). The result showed that BrApx isoforms were differentially expressed and the Br-chlApx, in particular in the layer, had the highest expression level and remained unchanged also after 14 day after harvest. In addition, expression changes were compared with total BrAPX activity and the results showed that the activity decreased in all tissues at 14 day after harvest, independently of transcripts. Finally, additional solutes as the substrate of APX ascorbate and its oxidized form, dehydroascorbate, as well as α-tocopherol, the major vitamin E compound that prevents the propagation of lipid peroxidation in thylakoid membranes, were followed. The changes in the BrApx expression, BrAPX activity and metabolites can provide further evidence of the close relationships that exist between antioxidants which compensate for each other and suggest that there are multiple sites of reciprocal control.


Effect of diluent sugars on capacitation status and acrosome reaction of spermatozoa in buck semen at refrigerated temperature.
The aim of the study was to explore the possibility of a better sugar suitable for storage of goat semen at refrigerated temperature. For this experiment, semen was collected from eight Jakhrana bucks maintained at Jakhrana unit, ICAR-CIRG, at twice a week interval using artificial vagina. Collected semen was preliminary evaluated, and better semen samples were pooled and divided into two parts. One part of the pooled semen was diluted in egg yolk, Tris, citric acid, and fructose diluter, whereas second part was diluted in egg yolk, Tris, citric acid, and glucose diluter. Then semen samples were kept in equilibration chamber for 4 h at 5 °C after proper dilution. Both the semen samples were evaluated for viability, motility, plasma membrane integrity, sperm abnormality, lipid peroxidation, acrosomal integrity, and capacitation status at 0 h, 24 h, 48 h, and 72 h after dilution. Significantly (P < 0.05) higher motility was observed at 24 h in extender containing glucose as compared with extender containing fructose but motility was decreased at 48 h and 72 h. Number of capacitated sperm increased significantly (P < 0.05) and acrosomal integrity was decreased significantly (P < 0.05) at 72 h in extender containing glucose. The other parameters like viability and plasma membrane integrity were decreased significantly (P < 0.05) at 72 h and lipid peroxidation as well as sperm abnormality increased significantly (P < 0.05) in extender containing glucose. From this study, it can be concluded that fructose is better diluent sugar for refrigerated storage of buck semen.


Enhanced registered nurse care coordination with sensor technology: Impact on length of stay and cost in aging in place housing.
When planning the Aging in Place Initiative at TigerPlace, it was envisioned that advances in technology research had the potential to enable early intervention in health changes that could assist in proactive management of health for older adults and potentially reduce costs. The purpose of this study was to compare length of stay (LOS) of residents living with environmentally embedded sensor systems since the development and implementation of automated health alerts at TigerPlace to LOS of those who are not living with sensor systems. Estimate potential savings of living with sensor systems. LOS for residents living with and without sensors was measured over a span of 4.8 years since the implementation of sensor-generated health alerts. The group living with sensors (n = 52) had an average LOS of 1,557 days (4.3 years); the comparison group without sensors (n = 81) was 936 days (2.6 years); p = .0006. Groups were comparable based on admission age, gender, number of chronic illnesses, SF12 physical health, SF12 mental health, Geriatric Depression Scale (GDS), activities of daily living, independent activities of daily living, and mini-mental status examination scores. Both groups, all residents living at TigerPlace since the implementation of health alerts, receive registered nurse (RN) care coordination as the standard of care. Results indicate that residents living with sensors were able to reside at TigerPlace 1.7 years longer than residents living without sensors, suggesting that proactive use of health alerts facilitates successful aging in place. Health alerts, generated by automated algorithms interpreting environmentally embedded sensor data, may enable care coordinators to assess and intervene on health status changes earlier than is possible in the absence of sensor-generated alerts. Comparison of LOS without sensors TigerPlace (2.6 years) with the national median in residential senior housing (1.8 years) may be attributable to the RN care coordination model at TigerPlace. Cost estimates comparing cost of living at TigerPlace with the sensor technology vs. nursing home reveal potential saving of about $30,000 per person. Potential cost savings to Medicaid funded nursing home (assuming the technology and care coordination were reimbursed) are estimated to be about $87,000 per person. Early alerts for potential health problems appear to enhance the current RN care coordination care delivery model at TigerPlace, increasing LOS for those living with sensors to nearly twice that of those who did not. Sensor technology with care coordination has cost saving potential for consumers and Medicaid.


Time- and cell-resolved dynamics of redox-sensitive Nrf2, HIF and NF-κB activities in 3D spheroids enriched for cancer stem cells.
Cancer cells have an altered redox status, with changes in intracellular signaling pathways. The knowledge of how such processes are regulated in 3D spheroids, being well-established tumor models, is limited. To approach this question we stably transfected HCT116 cells with a pTRAF reporter that enabled time- and cell-resolved activity monitoring of three redox-regulated transcription factors Nrf2, HIF and NF-κB in spheroids enriched for cancer stem cells. At the first day of spheroid formation, these transcription factors were activated and thereafter became repressed. After about a week, both HIF and Nrf2 were reactivated within the spheroid cores. Further amplifying HIF activation in spheroids by treatment with DMOG resulted in a dominant quiescent stem-cell-like phenotype, with high resistance to stress-inducing treatments. Auranofin, triggering oxidative stress and Nrf2 activation, had opposite effects with increased differentiation and proliferation. These novel high-resolution insights into spatiotemporal activation patterns demonstrate a striking coordination of redox regulated transcription factors within spheroids not occurring in conventional cell culture models.


[Experience in the removal of difficult and high risk tracheobronchial foreign body by bronchoscopy].
To explore the best methods and skill for the removal of difficult and high risk tracheobronchial foreign body under bronchoscope. A retrospective review was performed between August 1995 to August 2012. There were 4217 children with tracheobronchial foreign body, among them, 272 were diagnosed as high-risk, highly difficult tracheobronchial foreign bodies confirmed by clinical manifestations, foreign body type and bronchoscopy. In 271 children, the tracheobronchial foreign body was removed under bronchoscope, the success rate was 99.6%; only one child with a pen cap blocking the left lower lobe bronchus was transferred to the department of thoracic surgery, and the foreign body was finally removed by thoracotomy. Eighty-five children (among them, 82 children were under 1 year of age) had II-II degree laryngeal obstruction, the emergency surgery was performed to remove the foreign body and to relieve the laryngeal obstruction. Twenty-six children had lung infection and 27 children had failed foreign body removal surgery before, in all these children, the foreign body was removed after infection control. There were 17 children with the pen cap as the tracheobronchial foreign body, direct removal was successful in 12 children with the history less than two weeks; in 4 children, the foreign body was removed after 0.1% epinephrine saline flush, and 1 case with the homemade bronchial foreign body hook remove. There were 26 children with the whistle as the foreign body, and 32 children had large and sharp foreign bodies. In these cases, the foreign bodies were removed together with the bronchoscope. Forty-two children had multiple or fragile foreign bodies, and 16 children had subsegmental bronchial foreign bodies. In these cases, the foreign bodies were removed with forceps under direct vision and intraoperative bronchial lavage.In This series, 129 children received intraoperative bronchial lavage, among them, 127 children showed normal X-ray changes one week after operation. Two children with a history of more than 1 month complicated with pulmonary consolidation. After bronchial lavage, pneumothorax and subcutaneous emphysema occurred, which recovered after treatment. No glottic edema, asphyxia, and other complications were found, the complication rate of surgery was 0.7%. For the removal of highly difficult and high risk tracheobronchial foreign bodies, preoperative analysis and discussion should be sufficient, appropriate surgical skill and surgical instruments may improve the success rate of the surgery and prevent the operation complications.


Mortality among active-duty male French Armed Forces, 2006-10.
In the Armed Forces, knowledge about the causes of deaths is required in order to develop prevention strategies. This study presents the main characteristics of causes of deaths among male active-duty personnel in the French Armed Forces during the 2006-10 period and compares them with the general French male population. The data are provided by military public health surveillance. Comparisons of the specific mortality rates (MR) were performed using a Poisson regression. Standardized mortality ratios (SMRs) were calculated to compare mortality with the general French male population. There were 1455 deaths among male active-duty personnel during the study period [MR: 100.9 per 100,000 person-years (PY); 95% confidence interval 95.7-106.1]. The 17-24 age group was characterized by violent deaths: transport accident (MR: 45.9 per 100,000 PY) and suicide (18.8 per 100 000 PY). Overall SMRs show significantly lower MR compared with the French national MR with the exception of SMR for transport accident and suicide in the 17-24 age group. There is a significantly lower deficit of mortality compared with the French male general population, reflecting a strong healthy worker effect. However, health promotion programmes should continue to put emphasis on transport accident especially among the 17-24 age group.


Aflatoxin production by entomopathogenic isolates of Aspergillus parasiticus and Aspergillus flavus.
Fourteen isolates of Aspergillus parasiticus and 2 isolates of Aspergillus flavus isolated from the mealybug Saccharicoccus sacchari were analyzed for production of aflatoxins B1, B2, G1, and G2 in liquid culture over a 20-day period. Twelve Aspergillus isolates including 11 A. parasiticus and 1 A. flavus produced aflatoxins which were extracted from both the mycelium and culture filtrate. Aflatoxin production was detected at day 3 and was detected continually for up to day 20. Aflatoxin B1 production was greatest between 7 and 10 days and significantly higher quantities were produced by A. flavus compared to A. parasiticus. Aflatoxin production was not a stable trait in 1 A. parasiticus isolate passaged 50 times on agar. In addition to loss of aflatoxin production, an associated loss in sporulation ability was also observed in this passaged isolate, although it did maintain pathogenicity against S. sacchari. An aflatoxin B1 concentration of 0.16 micrograms/mealybug (14.2 micrograms/g wet wt) was detected within the tissues of infected mealybugs 7 days after inoculation. In conclusion, the ability of Aspergillus isolates to produce aflatoxins was not essential to the entomopathogenic activity of this fungus against its host S. sacchari.


Lectin-based impedimetric biosensor for differentiation of pathogenic candida species.
Fungi stand out as primary pathogens present in healthcare-acquired infections, presenting an increased number of cases even using appropriate antifungal therapy. Candida spp. is a predominant microorganism among several fungal pathogens present in the healthcare setting. Candidemia and candidiasis are fungal infections responsible for high morbidity and mortality among ill patients in hospitals. It is noticeable that prolonged hospital stays lead to a higher economic impact and increased risk for developing secondary fungal or even bacterial infections. New fast and sensitive approaches for the detection of Candida species is highly required. Electrochemical biosensors are an excellent alternative to conventional techniques by combining fast analyte detection, low cost, and the possibility of miniaturization. Lectins are carbohydrate-binding proteins with the capability to reach out to the microorganism cell wall. In this work, we proposed the development of an impedimetric biosensor for Candida spp. based on Concanavalin A (ConA) and wheat germ agglutinin (WGA) as recognition agents of the yeast cells. Atomic force microscopy images indicate changes in the biosensor surface after assembly of the molecules and exposure to fungal samples. Electrochemical impedance spectroscopy results revealed a proportional increase of charge transfer resistance (RCT) as fungal CFU increased, where four Candida species were evaluated (Candida krusei, Candida tropicalis, Candida parapsilosis and Candida albicans). The biosensor is useful to differentiate Candida spp. with a detection limit between 102 to 106 CFU mL-1. The obtained biosensor appears as an innovative candidate for the detection and differentiation of pathogenic Candida spp.


Correction to: Use of genomic information to exploit genotype-by-environment interactions for body weight of broiler chicken in bio-secure and production environments.
After publication of this work [1], we noticed that there was an error: the formula to calculate the standard error of the estimated correlation.


Dietary plant pigment on blood-digestive physiology, antioxidant-immune response, and inflammatory gene transcriptional regulation in spotted snakehead (Channa punctata) infected with Pseudomonas aeruginosa.
The current study addressed to investigate the effect of lycopene (LYC) on blood physiology, digestive-antioxidant enzyme activity, specific-nonspecific immune response, and inflammatory gene transcriptional regulation (cytokines, heat shock proteins, vitellogenins) in spotted snakehead (Channa punctata) against Pseudomonas aeruginosa. In unchallenged and challenged fish treated with 200 mg LYC enriched diet the growth performance and digestive-antioxidant enzymes increased after 30 days, whereas with inclusion of 100 or 400 mg LYC in the diets, the increase manifested on or after 45 days. No mortality in fish treated with any LYC diet against P. aeruginosa was revealed. In the unchallenged and challenged fish the phagocytic (PC) activity in head kidney (HK) and spleen were significantly enhanced when fed the control diet or other LYC diets, whereas the respiratory burst (RB) activity and nitric oxide (NO) production significantly increased when fed the 200 mg diet for 45 and 60 days. Similarly, the lysozyme (Lyz) activity in the HK and spleen, and total Ig content in serum were significantly higher in both groups fed the 200 mg LYC diet for 15, 45, and 60 days. Heat shock protein (Hsp 70) was significantly improved in the uninfected group fed the 200 mg LYC diet for 45 and 60 days, but Hsp27 did not significantly change among the experimental groups at any time points. TNF-α and IL-6 mRNA pro-inflammatory cytokine expression significantly increased in both groups fed the 200 mg LYC diet after 45 and 60 days, while the IL-12 mRNA expression was moderate in both groups fed the same diet for 60 days. The IL-10 did not significant mRNA expression between groups at any sampling. The iNOS and NF-κB mRNA expression was pointedly high in both groups fed the 200 mg LYC diet on day 45 and 60. Vitellogenin A (VgA) mRNA was significantly higher in the uninfected fish fed the 100 and 200 mg LYC diets for 45 and 60 days, but VgB did not reveal significant difference between the treatment groups at any time points. The present results suggest that supplementation of LYC at 200 mg significantly modulate the blood physiology, digestive-antioxidant enzymes, specific-nonspecific immune parameters, and cytokines, Hsp, and vitellogenins in spotted snakehead against P. aeruginosa.


White matter abnormalities in patients with treated hyperphenylalaninaemia: magnetic resonance relaxometry and proton spectroscopy findings.
In order to further clarify the pathogenesis and clinical significance of MRI white matter abnormalities in treated hyperphenylalaninaemia (HPA), ten patients (seven type I HPA, two type II and one type III) underwent T2 relaxometry (n = 8) and/or 1H spectroscopy (n = 7) in addition to conventional MR spin-echo imaging at 1.5 T. Two patients with severe MRI abnormalities had repeat examinations during and after a 6- to 8-month period of strict diet control. The clinical evaluation included a detailed neurological examination. In nine out of ten patients visual evoked potentials (VEP) were obtained parallel to the MR examination. MR imaging demonstrated typical symmetrical areas of prolonged T2 relaxation time predominantly in the posterior periventricular white matter in all but one of type I and II patients. There was no consistent relationship between MRI findings and time of diagnosis/initiation of therapy, IQ or visual evoked potential changes. MRI abnormalities tended to be more severe in patients with poor dietary control and high current plasma phenylalanine levels, whereas a normal MRI was found only in patients with plasma phenylalanine levels continuously below 0.36 mmol/l. There was marked regression of MRI abnormalities already after 3 months of strict diet control. T2 relaxometry showed a bi-exponential behaviour of T2 in the affected white matter, with a slow component of about 200-450 ms, indicating an increase in free (extracellular) water. 1H spectroscopy revealed no signs of severe neuronal damage. We conclude, that the observed white matter changes in treated HPA probably represent reversible structural myelin changes rather than permanent demyelination.


Plant immunity: AvrPto targets the frontline.
Bacterial pathogens must suppress host defences to cause disease. New research shows that the Pseudomonas effector protein AvrPto does so by directly targeting plant transmembrane receptor kinases involved in bacterial perception.


Upregulation of heart PFK-2/FBPase-2 isozyme in skeletal muscle after persistent contraction.
Fructose-2,6-bisphosphate (Fru-2,6-P(2)) is the most potent allosteric activator of liver 6-phosphofructo-1-kinase enzyme, which is crucial for glycolysis. It is present in skeletal muscle but its importance is controversial as a regulator of muscle glycolysis. This study aims to determine the role of Fru-2,6-P(2) in the control of muscle glycolysis during contraction. Muscle contraction was produced by chronic low-frequency stimulation of rabbit tibialis anterior for 24 h, followed by a rest period of 48 h. To determine muscle glycolysis adaptation, we applied a short functional electrostimulation test using the same system of low-frequency stimulation for 1, 3, and 10 s. The variation in concentration of lactate and pyruvate was used to calculate the flux along the glycolysis pathway and the Fru-1,6-P(2)/Fru-6-P ratio permitted to analyze the 6-phosphofructo-1-kinase activation. Fru-2,6-P(2) levels increased over the 24 h of stimulation and remained elevated after the rest period, this being the only metabolite that kept the changes produced by chronic low-frequency stimulation during the rest. During the short functional electrostimulation test, the glycolytic pathway in stimulated and rested muscle was more active than in control muscle, which coincided with higher kinase activity of the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2/FBPase-2) enzyme. Furthermore, we found a decrease in muscle, liver, and ubiquitous PFK-2/FBPase-2 isoform expression and an increase in heart isoform expression. For the first time, we demonstrate that a persistent increase in Fru-2,6-P(2) produced by a change in PFK-2/FBPase-2 isoform expression may play an important role in the regulation of muscle glycolysis during the first moments of exercise.


Surface nanoscale imaging of collagen thin films by Atomic Force Microscopy.
Collagen, the most abundant protein in mammals, due to its unique properties is widely used as biomaterial, scaffold and culture substrate for cell and tissue regeneration studies. Since the majority of biological reactions occur on surfaces and structures at the nanoscale level it is of great importance to image the nanostructural surface of collagen based materials. The aim of this paper was to characterize, with Atomic Force Microscopy (AFM), collagen thin films formed on different substrates (glass, mica, polystyrene latex particle surfaces) and correlate their morphology with the used substrates, formation methodologies (spin coating, hydrodynamic flow) and original collagen solution. The results demonstrated that, by altering a number of parameters, it was possible to control the formation of collagen nanostructured films consisting of naturally occurring fibrils. The spin coating procedure enabled the formation of films with random oriented fibrils, while substrates influenced the fibril packing and surface roughness. The hydrodynamic flow was used for guiding fibril major orientation, while adsorption time, rinsing with buffer and solution concentration influenced the fibril orientation. The clarification of the contribution that different parameters had on thin film formation will enable the design and control of collagen nanobiomaterials with pre-determined characteristics.


Asymmetric synthesis of cis- and trans-2,5-disubstituted pyrrolidines from 3-oxo pyrrolidine 2-phosphonates: synthesis of (+)-preussin and analogs.
Pyrrolidine enones, derived from 3-oxo pyrrolidine 2-phosphonates and a HWE reaction with aldehydes, on Luche reduction give pyrrolidine allylic alcohols. The alcohols on hydrogenation (Pd/H2) give cis-2,5-disubstituted pyrrolidines and on treatment with TFA-NaBH3CN undergo a hydroxy directed reduction to trans-2,5-disubstituted pyrrolidines.


Negative evidence of difference between right- and left-handers in interhemispheric transfer of information.
The present study has been designed to investigate the relationship between handedness and callosal function. Based on the previous finding of a greater anatomical connection between the hemispheres in left-handers than in right-handers and in order to test the possibility of greater functional communication, the efficiency of interhemispheric transfer of information has been measured using a finger localization task. Comparison of two groups of male right-handed and left-handed subjects shows no difference in the efficiency of interhemispheric transfer of information.


The enhancement of specific gene transcription in isolated nuclei by added HeLa whole cell extract.
Transcription was carried out in isolated rat liver nuclei by endogenous RNA polymerase with the addition of HeLa whole cell extract (HWCE) in the presence of nucleoside 5'-[gamma-S]triphosphate. The resulting 5'-gamma-thiophosphate on the synthesized RNA allows separation of in vitro initiated RNA from bulk RNA by mercury-agarose chromatography. HWCE not only increased initiation of new RNA chains greater than 3 times but also had no effect on the RNA synthesis by RNA polymerase II. The initiation of transcription of albumin and alpha-fetoprotein genes in an isolated nuclei system was selectively enhanced by HWCE. Using this system, we studied the effect of glucocorticoid on albumin and alpha-fetoprotein gene expression in vitro.


Inflammasome signalling in brain function and neurodegenerative disease.
The mammalian CNS is an intricate and fragile structure, which on one hand is open to change in order to store information, but on the other hand is vulnerable to damage from injury, pathogen invasion or neurodegeneration. During senescence and neurodegeneration, activation of the innate immune system can occur. Inflammasomes are signalling complexes that regulate cells of the immune system, which in the brain mainly includes microglial cells. In microglia, the NLRP3 (NOD-, LRR- and pyrin domain-containing 3) inflammasome becomes activated when these cells sense proteins such as misfolded or aggregated amyloid-β, α-synuclein and prion protein or superoxide dismutase, ATP and members of the complement pathway. Several other inflammasomes have been described in microglia and the other cells of the brain, including astrocytes and neurons, where their activation and subsequent caspase 1 cleavage contribute to disease development and progression.


Heterogeneity of the rhamnomannans from one strain of the human pathogen Sporothrix schenckii determined by 13C nuclear magnetic resonance spectroscopy.
The synthesis of different rhamnomannans in a strain of Sporothrix schenckii (1099.12) was shown by use of 13C nuclear magnetic resonance spectroscopy. Fractionation of a polysaccharide preparation from cells grown at 25 degrees C provided a neutral monorhamnosyl rhamnomannan and an acidic rhamnomannan containing 4-O-substituted glucuronic acid units and also (1 leads to 2)-linked dirhamnosyl side chains.


Entry of feline calicivirus is dependent on clathrin-mediated endocytosis and acidification in endosomes.
Feline calicivirus is a major causative agent of respiratory disease in cats. It is also one of the few cultivatable members of Caliciviridae. We have examined the entry process of feline calicivirus (FCV). An earlier study demonstrated that acidification in endosomes may be required. We have confirmed this observation and expanded upon it, demonstrating, using drugs to inhibit the various endocytic pathways and dominant-negative mutants, that FCV infects cells via clathrin-mediated endocytosis. We have also observed that FCV permeabilizes cell membranes early during infection to allow the co-entry of toxins such as alpha-sarcin. Inhibitors of endosome acidification such as chloroquine and bafilomycin A1 blocked this permeabilization event, demonstrating that acidification is required for uncoating of the genome and access to the cytoplasm.


Organophosphate ester flame retardants in Nepalese soil: Spatial distribution, source apportionment and air-soil exchange assessment.
Despite soil being the major terrestrial environmental reservoir and one of the significant sinks for many hydrophobic organic compounds including organophosphate ester flame retardants (OPFRs), limited information is available about concentration and fate of OPFRs contamination in urban soil in general and especially in case of Nepal. This study investigates the environmental concentration, spatial distribution and source apportionment of eight OPFRs in surface soil (n = 28) from four major cities of Nepal with special interest on air-soil exchange. Overall, significantly high concentrations of ∑8OPFR were measured in soil ranging from 25-27,900 ng/g dw (median 248 ng/g dw). In terms of compositional pattern, tris(methyl phenyl) phosphate (TMPP) was the most abundant phosphorus chemical in soil, followed by tris(2-chloroisopropyl) phosphate (TCIPP), and accounted for 35-49% and 8-25% of ∑8OPFRs, respectively. The high level of these OPFRs was attributed to local sources as opposed to transboundary influence from remote areas. A Spearman's rank correlation analysis exhibited weak correlation of ∑8OPFRs with TOC (Rho = 0.117, p < 0.05) and BC (Rho = 0.007, p < 0.05), suggesting little or no influence of TOC and BC on the concentration of ∑8OPFRs. The fugacity fraction (ff) results indicated a strong influence of soil contamination on atmospheric level of OPFRs via volatilization.


The computer in clinical research. Prevalence of glaucoma.
The prevalence of glaucoma in a rural population of 6232 people over the age of 40 years, including patients with previously diagnosed glaucoma, was found to be 1.21%. There were 41 cases of open-angle glaucoma compared to 32 cases of angle-closure glaucoma. The prevalence of glaucoma among subjects over the age of 60 years was 3.07%.


Effectiveness of steam autoclaving on the contents of sharps containers.
The effectiveness of steam autoclaving on bacterial endospores placed within five types of sharps containers was tested. A variety of container physical orientations within the autoclave were evaluated. Spores were present on commercial spore strips or placed onto capped and uncapped dental needles. All strips and needles present in empty or filled containers could be sterilized within 15 minutes when the containers were placed on their sides and their vents left open. The contents of containers processed in an upward position required between 30-60 minutes of autoclaving before being sterilized. The size and shape of the containers influenced ease of sterilization.


Mid-season gas exchange of an alpine grassland under elevated CO2.
Ecosystem net CO2 uptake, evapotranspiration (ET) and night-time CO2 efflux were measured in an alpine grassland dominated by Carex curvula, treated with doubled ambient partial pressure of CO2 via open-top chambers. One quarter of the plots were treated with mineral nutrients to simulate the effect of lowland nitrogen deposition rates. Depending upon fertilizer supply, ecosystem net CO2 uptake per ground area in full sunlight (NCEmax) was 41-81% higher in open-top chambers supplied with doubled ambient partial pressure (p a) of CO2 than in plots receiving ambient CO2. Short-term reversals of the CO2 level suggest that the extent of downward adjustment of canopy photosynthesis under elevated CO2 was 30-40%. ET tended to decline, while water use efficiency (WUE), expressed as the NCEmax:ET ratio, increased more than twofold under elevated CO2. Night-time ecosystem CO2 efflux did not respond to changes in CO2 p a. NCEmax and night-time CO2 efflux were more responsive to mineral fertilizer than the doubling of CO2. This suggests that in these alpine plant communities, atmospheric nutrient input may induce equal or greater effects on gas exchange than increased CO2.


Risperidone-associated adverse drug reactions and CYP2D6 polymorphisms in a South African cohort.
Contradictory information exists regarding the influence of CYP2D6 polymorphisms on adverse drug reactions (ADRs) (extrapyramidal symptoms (EPS) and weight gain) related to risperidone treatment. This prompted us to evaluate the influence of CYP2D6 genetic variation in a cohort of South African patients who presented with marked movement disorders and/or weight gain while on risperidone treatment. Patients who were experiencing marked risperidone ADRs were recruited from Weskoppies Public Psychiatric Hospital. As poor or intermediate metabolism was expected, comprehensive CYP2D6 sequence variations were evaluated using XL-PCR + Sequencing. No statistically significant association was found between CYP2D6 poor metabolism and risperidone ADRs. An inverse relationship between EPS and weight gain was however identified. A novel CYP2D6 allele was identified which is unlikely to affect metabolism based on in silico evaluation. CYP2D6 variation appeared not to be a good pharmacogenetic marker for predicting risperidone-related ADRs in this naturalistic South African cohort. Evaluation of a larger cohort would be needed to confirm these observations, including an examination of the role of potential intermediaries between the hypothesised genetic and clinical phenotypes.


A comparative study of the relative sensitivity and specificity of radiolabelled monoclonal antibody and computerised tomography in the detection of sites of disease in human malignant melanoma.
A monoclonal antibody raised against the high molecular weight melanoma antigen was labelled with indium-111 and injected intravenously into 25 patients with malignant melanoma. The results obtained from images at 24 and 96 h post i.v. administration of the antibody were compared with results obtained from computerised tomography studies with regard to detection of previously unrecognised sites of metastatic disease and apparent false positive localisation. Detailed study of the patients' clinical condition and detection rates using the two methods suggest that both methods detect approximately 80% of clinically and pathologically confirmed metastases. Of 62 known metastases, the antibody detected 50 (81%), with 17 false positive results. False negatives were most common in the lung. In eight patients the two methods were considered of equal value, in 10 the monoclonal gave a greater amount of clinically relevant information, and in seven the CT was superior. In three patients clinically significant metastatic lesions were detected by the radiolabelled monoclonal and had not been previously recognised either by CT scanning or on clinical grounds. No patients had any adverse reaction to the antibody and in the course of our study the dose of antibody was reduced from 20 mg to 200 micrograms with no apparent loss of sensitivity. In at least two patients uptake of the labelled monoclonal into tumour sites would have been adequate for effective targeted radiotherapy.


Duodenal-jejunal bypass liners: outcomes in glycaemic control and weight loss.
Duodenal-jejunal bypass liners (DJBLs) are a novel therapy with potential treating a range of metabolic diseases. This review discusses the weight loss and glycaemic improvements observed following the use of the DJBL. We summarize the evidence for the clinical application of DJBL to date. Within this, we outline the evidence available on the mechanisms of the observed effects. Based on the most recent trials, we cannot fully mimic the clinical or physiological effects of bariatric surgery with this device, but we are getting closer.


Heterogeneity of atherosclerotic plaque characteristics in human coronary artery disease: a three-dimensional intravascular ultrasound study.
The objective of this study was to describe the intraplaque variability of coronary atherosclerosis in humans. Atherosclerosis is a heterogeneous process. The degree and patterns of intraplaque heterogeneity are not well described. This study uses 3D intravascular ultrasound (IVUS) to examine variability in individual atherosclerotic plaques in human coronary arteries. IVUS images of 170 coronary plaques in 98 patients were evaluated. Each plaque was divided into proximal, middle, and distal sections. Quantitative and qualitative analyses were performed for each section using a dedicated 3D IVUS protocol. Intralesion heterogeneity was assessed between sections. Heterogeneity in composition was observed in most plaques (89%). The pattern of remodeling was heterogeneous in 23% of lesions. External elastic membrane (EEM) areas demonstrated an average percent deviation of 28.9% +/- 15.5%. Positive remodeling was associated with longer lesions (>> median length of 12.7 mm) (P = 0.031). Soft and calcific sections had a smaller mean EEM area (P = 0.034). Calcific lesions had a smaller mean lumen area (P = 0.027) and a greater percent plaque burden (PPB) (P = 0.001). Neither the location within the vessel or within the plaque was associated with plaque morphology. Greater qualitative heterogeneity was found in patients presenting with acute coronary syndrome (P < 0.001). Our results demonstrate a high degree of heterogeneity in composition and morphological features within individual atherosclerotic plaques in human coronary arteries. Intraplaque heterogeneity represents a challenge for imaging protocols correlating plaque features with cardiovascular events and for the development of future therapeutic options.


Pharmacokinetics of sulfaethidole in the rat: nonlinear multicompartment solution.
Sulfaethidole distribution and elimination in the rat was studied over a 90-fold dose range. This experimental design produced marked nonlinearity in the binding of sulfaethidole to proteins in both interstitial fluid an plasma. Using a multicompartmental model consisting of binding of sulfaethidole to plasma and interstitial fluid proteins, sulfaethidole distribution in the body could be simulated. Urinary and biliary elimination of sulfaethidole depended on the unbound drug mass in the plasma and urine flow. The results confirm the central role of the unbound species in the distribution and elimination of drugs with marked binding to plasma proteins.


Recent findings about the Yersinia enterocolitica phage shock protein response.
The phage shock protein (Psp) system is a conserved extracytoplasmic stress response in bacteria that is essential for virulence of the human pathogen Yersinia enterocolitica. This article summarizes some recent findings about Y. enterocolitica Psp system function. Increased psp gene expression requires the transcription factor PspF, but under non-inducing conditions PspF is inhibited by an interaction with another protein, PspA, in the cytoplasm. A Psp-inducing stimulus causes PspA to relocate to the cytoplasmic membrane, freeing PspF to induce psp gene expression. This PspA relocation requires the integral cytoplasmic membrane proteins, PspB and PspC, which might sense an inducing trigger and sequester PspA by direct interaction. The subsequent induction of psp gene expression increases the PspA concentration, which also allows it to contact the membrane directly, perhaps for its physiological function. Mutational analysis of the PspB and PspC proteins has revealed that they both positively and negatively regulate psp gene expression and has also identified PspC domains associated with each function. We also compare the contrasting physiological roles of the Psp system in the virulence of Y. enterocolitica and Salmonella enterica sv. Typhimurium (S. Typhimurium). In S. Typhimurium, PspA maintains the proton motive force, which provides the energy needed to drive ion importers required for survival within macrophages. In contrast, in the extracellular pathogen Y. enterocolitica, PspB and PspC, but not PspA, are the Psp components needed for virulence. PspBC protect Y. enterocolitica from damage caused by the secretin component of its type 3 secretion system, an essential virulence factor.


Total body wash-out in patients with fulminant hepatitis.
Two patients with fulminant hepatic necrosis and in stage IV coma were treated with a rapid infusion of refrigerated buffered albumin electrolyte solution combined with simultaneous total body wash-out in order to remove the circulating neurotoxins and to provide additional time for liver regeneration. Both patients awoke from the coma following treatment. One patient recovered completely, the other died on the second day as a result of massive gastro-intestinal bleeding. The clinical value and the technical problems of the procedure are investigated.


Maintaining Superior Follow-Up Rates in a Longitudinal Study: Experiences from the College Life Study.
Longitudinal studies are often considered to be a gold standard for research, but the operational management of such studies is not often discussed in detail; this paper describes strategies used to track and maintain high levels of participation in a longitudinal study involving annual personal interviews with a cohort of 1,253 undergraduates (first-time, first-year students at time of enrollment) at a large public mid-Atlantic university.


Interaction mechanism of Trp-Arg dipeptide with calf thymus DNA.
The interaction between Trp-Arg dipetide (WR) and calf thymus DNA (ctDNA) in pH 7.4 Tris-HCl buffer was investigated by multi-spectroscopic techniques and molecular modeling. The fluorescence spectroscopy and UV absorption spectroscopy indicated that WR interacted with ctDNA in a minor groove binding mode and the binding constant was 4.1 × 10(3). The ionic strength effect and single-stranded DNA (ssDNA) quenching effect further verified the minor groove binding mode. Circular dichroism spectroscopy (CD) was employed to measure the conformation change of ctDNA in the presence of WR. The molecular modeling results illustrated that electrostatic interaction and groove binding coexisted between them and the hydrogen bond and Van der Waals were main acting forces. All the above methods can be widely used to investigate the interaction of peptide with nucleic acids, which contributes to design the structure of new and efficient drugs.


Effect of enoxaparin and aspirin on hemodynamic disturbances after global cerebral ischemia in rats.
Whether patients benefit from anticoagulation or anti-platelet agents when subjected to global ischemia is controversial. In this study we assessed the effect of enoxaparin and aspirin on hemodynamic disturbances following global ischemia in rats. Twenty-minute global ischemia was induced by four-vessel occlusion in Sprague-Dawley rats. Enoxaparin (1.5 mg/kg, i.v.) was administered 4 min before or after ischemia, and aspirin (40 mg/kg, i.p.) was administered twice, 18 and 1.5 h before ischemia. Parietal cortical cerebral blood flow was measured in real time with a laser Doppler flowmeter, and activated partial thromboplastin time (APTT) and prothrombin time (PT) were measured. Three phenomena of hemodynamic disturbance: no-reflow, hyperperfusion, and hypoperfusion, were assessed after 20-min global ischemia. No significant differences in PT values were found; in contrast, the APTT values were shortened markedly by 20-min ischemia, which indicated activation of the endogenous coagulation system. The shortened APTT was markedly prevented by enoxaparin administered before ischemia. No-reflow was partly mitigated by administration of enoxaparin or aspirin before ischemia. Hypoperfusion was almost fully blocked by aspirin, and the hypoperfusion value was increased and hypoperfusion duration was shortened by enoxaparin administered 4 min before or after ischemia. These results indicate that enoxaparin and aspirin have protective effects against cerebral hemodynamic disturbances induced by global ischemia in rats.


Local administration of flurazepam has different effects on dopamine release in striatum and nucleus accumbens: a microdialysis study.
The action of local administration of flurazepam on extracellular levels of dopamine (DA) and its metabolites 3,4 dihydroxyphenyl acetic acid (DOPAC) and homovanillic acid (HVA) in the anterior striatum and medial nucleus accumbens have been investigated using microdialysis. Flurazepam (10 microM), administered through the perfusion medium for 20 min, reduced levels of DA in dialysates from the nucleus accumbens by 60% while the same concentration of the drug had no effect on levels of DA in perfusates from the striatum. Pretreatment with the benzodiazepine antagonist Ro 15-1788 (flumazenil) or with picrotoxin, a drug which blocks the GABAA receptor-associated chloride channel, inhibited the effect of flurazepam on levels of DA in the nucleus accumbens, which suggests that the effect was mediated by the multimolecular GABAA/benzodiazepine receptor complex. Administration of flurazepam had little effect on the two metabolites of DA, DOPAC and HVA, in either the nucleus accumbens or striatum. The inverse partial benzodiazepine agonist, FG 7142, had no significant action on the release of DA in the nucleus accumbens. These results suggest that the dopaminergic projection to the nucleus accumbens is more sensitive to benzodiazepine-induced inhibition than the projection to the striatum.


Prognostic Impact of Using Combined Plasma Fibrinogen Level and Neutrophil-to-Lymphocyte Ratio in Resectable Non-small Cell Lung Cancer.
Both plasma fibrinogen level and neutrophil-to-lymphocyte ratio (NLR) are associated with malignant potential in different cancer types. The current study evaluated the use of combined plasma fibrinogen level and NLR (F-NLR) as a prognostic predictor in patients with non-small cell lung cancer (NSCLC). Data collected from 279 patients with resectable NSCLC were retrospectively reviewed. Patients were divided into three groups based on the F-NLR score: score 2, high fibrinogen level (≥350 mg/dL) and high NLR (≥2.5); score 1, either high fibrinogen level or high NLR; and score 0, neither abnormal. Overall survival (OS) and relapse-free survival (RFS) were evaluated using the Kaplan-Meier method and log-rank test. Cox proportional hazard model was used to assess prognostic factors. Numbers of patients with F-NLR score of 0, 1, and 2 were 122 (43.7%), 105 (37.6%), and 52 (18.6%), respectively. The F-NLR was found to be significantly associated with age, male sex, heavy smoking history, high pT status and pathological stage, and nonadenocarcinoma. Moreover, the OS and RFS significantly differed according to the F-NLR score (P < 0.001, P = 0.003). A multivariate analysis revealed that a high F-NLR score (≥1) was an independent poor prognostic factor for OS (P = 0.027). In subgroup analyses, an adverse prognostic impact of the F-NLR score on OS was identified regardless of nodal involvement or pathological stage. The F-NLR score, which is based on histological inflammation and coagulability, could be a potential prognostic indicator in patients with resectable NSCLC.


Urinary high density lipoprotein in minimal change glomerular disease and chronic glomerulopathies.
Serum lipids and lipoproteins and urinary apolipoprotein A (Apo A) were determined in two groups of patients. One group consisted of 11 children (ages ranging from 4 to 14 years) with minimal change glomerular disease. The other group consisted of 13 patients, eight less than 19 years old five adults, with different types of chronic glomerulopathy. Elimination of urinary lysozyme was a feature of chronic glomerulopathies, and creatinine clearances were also significantly lower in this group. Patients with chronic glomerulopathies had significantly lower HDL cholesterol and Apo A concentrations in their sera. In contrast, urinary Apo A concentrations were significantly higher in patients with chronic glomerulopathies, who also showed significantly lower urinary protein selectivities. Lipoprotein electrophoresis of urines containing Apo A showed distinct high-density lipoprotein (HDL) fractions, suggesting that HDL is eliminated in the urine as a result of increased glomerular permeability. This is also supported by a correlation coefficient of 0.77 between the selectivity indices and the ratio of urinary Apo A to total proteinuria. The determination of urinary Apo A appears to give valuable diagnostic information in patients with glomerular disease. According to our results the absence of urinary Apo A is very suggestive of minimal change glomerular disease.


Optimal HSF1 activation in response to acute cold stress in BAT requires nuclear TXNIP.
While TXNIP (thioredoxin interacting protein) in the plasma membrane and vesicular location is known to negatively regulate cellular glucose uptake by facilitating glucose transporter endocytosis, the function of TXNIP in the nucleus is far less understood. Herein, we sought to determine the function of nuclear TXNIP in vivo, using a new HA-tagged TXNIP knock-in mouse model. We observed that TXNIP can be found in the nucleus of a variety of cells from different tissues including hepatocytes (liver), enterocytes (small intestine), exocrine cells (pancreas), and brown adipocytes (BAT). Further investigations into the role of nuclear TXNIP in BAT revealed that cold stress rapidly and transiently activated HSF1 (heat shock factor 1). HSF1 interaction with TXNIP during its activation is required for optimal HSF1 directed cold shock response in BAT.


Preliminary proteomic analysis of radiation response markers in rectal cancer patients.
To provide biomarkers related to the radiation response of patients to avoid unnecessary side effects on those who were not sensitive to radiotherapy. In the present study, we compared the different four proteins (PDIA3, Vimentin, Galectin3, Dhe3) patterns in rectal tumor tissue before and after radiation therapy by using 2-D PAGE, mass spectrometry, and bioinformatics analysis. The protein level of Galectin3 and PDIA3 were downregulated in rectal cancer patients before and after radiotherapy (1.42 folds); while Dhe3 protein and Vimentin were upregulated (1-2 folds), and we also revealed Vimentin as its role in the negative regulation of the well-known transcription factor ATF4. Our study showed that four candidate proteins, including PIDA3, Galectin3, Dhe3, and Vimentin, might be the potential biomarkers in the identification of radiation response in rectal cancer.


Cost-effectiveness analysis of pharmacologic treatment of fibromyalgia in Mexico.
To identify, from the Mexican Public Health System perspective, which would be the most cost-effective treatment for patients with Fibromyalgia (FM). A Markov model including three health states, divided by pain intensity (absence or presence of mild, moderate or severe pain) and considering three-month cycles; costs and effectiveness were estimated for amitriptyline (50mg/day), fluoxetine (80 mg/day), duloxetine (120 mg/day), gabapentin (900 mg/day), pregabalin (450 mg/day), tramadol/acetaminophen (150 mg/1300 mg/día) and amitriptyline/fluoxetine (50mg/80 mg/día) for the treatment of FM. The clinical outcome considered was the annual rate of pain control. Probabilities assigned to the model were collected from published literature. Direct medical costs for FM treatment were retrieved from the 2006 data of the Mexican Institute of Social Security (IMSS) databases and were expressed in 2010 Mexican Pesos. Probabilistic Sensitivity Analyses were conducted. The best pain control rate was obtained with pregabalin (44.8%), followed by gabapentin (38.1%) and duloxetine (34.2%). The lowest treatment costs was for amitriptyline ($ 9047.01), followed by fluoxetine ($ 10,183.89) and amitriptyline/fluoxetine ($ 10,866.01). By comparing pregabalin vs amitriptyline, additional annual cost per patient for pain control would be around $ 50.000 and $ 75.000 and would result cost-effective in 70% and 80% of all cases. Among all treatment options for FM, pregabalin achieved the highest pain control and was cost-effective in 80% of patients of the Mexican Public Health System.


Approach to Diagnosis and Management of Treatment-Resistant Depression.
Major depressive disorder is a chronic and recurrent illness that affects 20% of adults during their lifetime and is one of the leading causes of suicide in the United States. A systematic measurement-based care approach is the essential first step in the diagnosis and management of treatment-resistant depression (TRD) by promptly identifying individuals with depression and avoiding delays in treatment initiation. As comorbidities may be associated with poorer outcomes to commonly used antidepressants and increase risk of drug-drug interactions, their recognition and treatment is an essential component of management of TRD.


Comparative antioxidant activity and untargeted metabolomic analyses of cherry extracts of two Chinese cherry species based on UPLC-QTOF/MS and machine learning algorithms.
P. pseudocerasus and P. tomentosa are the two native Chinese cherry species of high economic and ornamental worths. Little is known about the metabolic information of P. pseudocerasus and P. tomentosa. Effective means are lacking for distinguishing these two similar species. In this study, the differences in total phenolic content (TPC), total flavonoid content (TFC), and in vitro antioxidant activities in 21 batches of two species of cherries were compared. A comparative UPLC-QTOF/MS-based metabolomics coupled with three machine learning algorithms was established for differentiating the cherry species. The results demonstrated that P. tomentosa had higher TPC and TFC with average content differences of 12.07 times and 39.30 times, respectively, and depicted better antioxidant activity. Total of 104 differential compounds were identified by UPLC-QTOF/MS metabolomics. The major differential compounds were flavonoids, organooxygen compounds, and cinnamic acids and derivatives. Correlation analysis revealed differences in flavonoids content such as procyanidin B1 or isomer and (Epi)catechin. They could be responsible for differences in antioxidant activities between the two species. Among three machine learning algorithms, the prediction accuracy of support vector machine (SVM) was 85.7%, and those of random forest (RF) and back propagation neural network (BPNN) were 100%. BPNN exhibited better classification performance and higher prediction rate for all testing set samples than those of RF. The study herein found that P. tomentosa had higher nutritional value and biological functions, and thus considered for usage in health products. Machine models based on untargeted metabolomics can be effective tools for distinguishing these two species.


Odontogenic Infections.
The pathogenesis of odontogenic infection is polymicrobial, consisting of various facultative and strict anaerobes. The dominant isolates are strictly anaerobic gram-negative rods and gram-positive cocci. The periapical infection is the most common form of odontogenic infection. Although odontogenic infections are usually confined to the alveolar ridge vicinity, they can spread into deep fascial spaces. Cavernous sinus thrombosis, brain abscess, airway obstruction, and mediastinitis are possible complications of dental infections. The most important element in treating odontogenic infections is elimination of the primary source of the infection with antibiotics as adjunctive therapy.


Insights into the molecular interactions between aminopeptidase and amyloid beta peptide using molecular modeling techniques.
Amyloid beta (Aβ) peptides play a central role in the pathogenesis of Alzheimer's disease. The accumulation of Aβ peptides in AD brain was caused due to overproduction or insufficient clearance and defects in the proteolytic degradation of Aβ peptides. Hence, Aβ peptide degradation could be a promising therapeutic approach in AD treatment. Recent experimental report suggests that aminopeptidase from Streptomyces griseus KK565 (SGAK) can degrade Aβ peptides but the interactive residues are yet to be known in detail at the atomic level. Hence, we developed the three-dimensional model of aminopeptidase (SGAK) using SWISS-MODEL, Geno3D and MODELLER. Model built by MODELLER was used for further studies. Molecular docking was performed between aminopeptidase (SGAK) with wild-type and mutated Aβ peptides. The docked complex of aminopeptidase (SGAK) and wild-type Aβ peptide (1IYT.pdb) shows more stability than the other complexes. Molecular docking and MD simulation results revealed that the residues His93, Asp105, Glu139, Glu140, Asp168 and His255 are involved in the hydrogen bonding with Aβ peptide and zinc ions. The interactions between carboxyl oxygen atoms of Glu139 of aminopeptidase (SGAK) with water molecule suggest that the Glu139 may be involved in the nucleophilic attack on Ala2-Glu3 peptide bond of Aβ peptide. Hence, amino acid Glu139 of aminopeptidase (SGAK) might play an important role to degrade Aβ peptides, a causative agent of Alzheimer's disease.