Allergen-specific regulation of allergic rhinitis in mice by intranasal exposure to IgG1 monoclonal antibody Fab fragments against pathogenic allergen.
Fab fragments (Fabs) have the ability to bind to specific antigens but lack the Fc portion for binding to receptors on immune and inflammatory cells that play a critical role in allergic diseases. In the present study, we investigated whether Fabs of an allergen-specific IgG1 monoclonal antibody (mAb) inhibited allergic rhinitis in mice. BALB/c mice sensitized by intraperitoneal injections of ovalbumin (OVA) plus alum on days 0 and 14 were intranasally challenged with OVA on days 28-30, and 35. Fabs prepared by the digestion of an anti-OVA IgG1 mAb (O1-10) with papain were also intranasally administered 15min before each OVA challenge. The results showed that treatment with O1-10 Fabs significantly suppressed the sneezing frequency, associated with decrease of OVA-specific IgE in the serum and infiltration by mast cells in the nasal mucosa seen following the fourth antigenic challenge; additionally, the level of mouse mast cell protease-1, a marker of mast cell activation, in serum was decreased. Furthermore, infiltration of eosinophils and goblet cell hyperplasia in the nasal mucosa at the fourth challenge were inhibited by treatment with O1-10 Fabs. In conclusion, these results suggest that intranasal exposure to Fabs of a pathogenic antigen-specific IgG1 mAb may be effective in regulating allergic rhinitis through allergen capture by Fabs in the nasal mucosa before the interaction of the intact antibody and allergen.


Topical Application of Keratinocyte Growth Factor Conjugated Gold Nanoparticles Accelerate Wound Healing.
Keratinocyte growth factor (KGF) has been demonstrated to specifically stimulate the multiplication and migration of keratinocytes. However, due to rapid degradation, the results of topical application of growth factors on wounds are unsatisfactory. In this study, we cross-linked KGF to the surface of gold nanoparticles (GNPs) and explored their effects on wound healing. The as-synthesized nanocomposite (KGF-GNPs) displayed good colloidal stability, decent biocompatibility as well as negligible cellular cytotoxicity. The in vitro cellular experimental results demonstrated that KGF-GNPs could effectively promote the proliferation of keratinocytes in contrast to bare GNPs or KGF. Furthermore, in animal full-thickness wound model, KGF-GNPs are more conducive to wound healing than bare GNPs or KGF. KGF-GNPs enhanced wound healing by promoting wound re-epithelialization rather than granulation. The superior biocompatibility, colloidal depressiveness and biological activity of this nanocomposite indicate that it could be utilized as a promising wound healing drug for clinical application in the future.


Reuse of effluent water from a municipal wastewater treatment plant in microalgae cultivation for biofuel production.
This study assessed the usability of effluent water discharged from a secondary municipal wastewater treatment plant for mass cultivation of microalgae for biofuel production. It was observed that bacteria and protozoa in the effluent water exerted a negative impact on the growth of Chlorella sp. 227. To reduce the effect, filtration or UV-radiation were applied on the effluent water as pre-treatment methods. Of all the pretreatment options tested, the filtration (by 0.2 μm) resulted in the highest biomass and lipid productivity. To be comparable with the growth in the autoclaved effluent water, the filtration with a proper pore size filter (less than 0.45 μm) or UV-B radiation of a proper dose (over 1620 mJ cm(-2)) are proposed. These findings led us to conclude that the utilization can be realized only when bacteria and other microorganisms are greatly reduced or eliminated from the effluent prior to its use.


A two-step hybrid technique for accurately localizing acoustic source in anisotropic structures without knowing their material properties.
Acoustic source localization techniques generally assume straight line propagation of waves from the acoustic source to the sensor. However, it is well-known that in anisotropic plates the acoustic energy does not always propagate in straight lines. Even for isotropic plates containing a cavity or an inclusion between the acoustic source and the sensor the straight line propagation assumption is violated. In such cases only options available in localizing acoustic source is to use relatively expensive distributed sensor systems, or to follow time reversal techniques based on the impulse response functions which is labor intensive and computationally demanding. A two-step hybrid technique is proposed in this paper for predicting acoustic source in anisotropic plates. During the first step it was assumed that the waves propagated along straight lines from the acoustic source to the sensor. The source was localized with this simplifying assumption. Then this first prediction was improved in the second step by solving an optimization problem. Experimental results showed that the second step always moved the estimates towards the actual source location. Thus it always reduced the prediction error irrespective of whether the final prediction coincided with the actual source location or not.


No unleashed expression without language.
While the metarepresentational structure of ostensive communication may explain the unleashing of human expression, it neither explains the open-endedness of the thoughts expressed/communicated, nor how the multiply embedded nature of the metarepresentational structure invoked arose. These both require the recursivity of human language, a capacity which must be distinguished from external (public) languages and their use in communication.


Analysis of the conformational space and dynamics of RNA helicases by single-molecule FRET in solution and on surfaces.
RNA helicases are a diverse group of enzymes that catalyze the unwinding of RNA duplex regions in an ATP-dependent reaction. Both the helicase itself and its RNA substrate undergo conformational changes during the reaction, which are amenable to Förster resonance energy transfer (FRET) studies. Single-molecule FRET studies in solution by confocal microscopy and on surfaces by total internal reflection microscopy provide information on different conformers present, their fractional populations in equilibrium, and the rate constants of their inter-conversion. Collectively, the information gained can be integrated into a kinetic and thermodynamic framework that quantitatively describes the conformational dynamics of the helicase studied. FRET experiments also provide distance information to map and model the structures of individual conformational states. The integrated model provides a comprehensive description of the structure and dynamics of the helicase, which can be linked to its biological function. Single-molecule FRET studies have tremendous potential to define the relationship between structure, function and dynamics of RNA helicases and to understand the mechanistic basis for their broad range of biological functions. The focus of this chapter is on providing guidance in the design of single-molecule FRET experiments and on the interpretation of the data obtained. Selected examples illustrate important considerations when analyzing single-molecule experiments, as well as their limitations and possible pitfalls.


Biology of basal cell carcinoma (Part II).
Host-tumor relationships involve several factors that can enhance or suppress neoplastic growth. This second part of a review of basal cell carcinoma biology examines the role that hormones, cytokines, local and systemic immunity, congenital and genetic syndromes, and environmental factors play in the development of this neoplasm. Theories of etiology and pathogenesis are discussed, and transplantation and cell culture techniques used to study this cancer are explored. Valuable second-line therapies for treatment of multiple tumors are reviewed, and important areas of present and future research are emphasized.


Fatty acids differentially influence phosphatidylinositol 3-kinase signal transduction in endothelial cells: impact on adhesion and apoptosis.
In contrast to n-6 fatty acids like arachidonic acid (AA), the anti-inflammatory potential of n-3 fatty acids such as docosahexaenoic acid (DHA) has been demonstrated. We examined the phosphatidylinositol (PI)3-kinase dependent effects of AA versus DHA on monocyte rolling, adhesion and transmigration through inflammatory activated human umbilical venous endothelial cells (HUVEC) as well as on apoptosis, to investigate the impact on vascular inflammation. HUVEC were pre-incubated with AA, DHA or sham, and stimulated with VEGF, TNF-alpha or staurosporine. Rolling and adhesion were investigated by means of a parallel flow chamber; transmigration was performed in a static assay. Activation of PI3-kinase was measured as phosphorylation of protein kinase B (Akt). Apoptosis was determined by caspase-3 activity and annexin-V analysis. Pre-incubation of HUVEC with DHA markedly decreased TNF-alpha-induced monocyte rolling, adhesion, and transmigration, although expression of endothelial adhesion molecules was unchanged. In contrast, AA increased TNF-alpha-induced rolling. Both fatty acids did not alter TNF-alpha-mediated upregulation of the adhesion molecules ICAM-1, VCAM-1, and E-selectin. The divergent effects of AA and DHA were abrogated with PI3-kinase inhibitors. After pre-incubation with DHA, VEGF-, TNF-alpha- and staurosporine-induced phosphorylation of Akt was decreased when compared to AA. DHA pre-incubation significantly increased staurosporine-induced apoptosis. In addition, DHA in comparison to AA augmented staurosporine-mediated increase in caspase-3 activity. In conclusion, DHA-induced a reduction in rolling, adhesion and transmigration of monocytes through inflammatory activated HUVEC that is in part PI3-kinase dependent. PI3-kinase driven phosphorylation of Akt and apoptosis of HUVEC as contribution to the resolution of inflammation is differentially modulated by DHA versus AA.


A complex interaction between glycine/NMDA receptors and serotonergic/noradrenergic antidepressants in the forced swim test in mice.
Both clinical and preclinical studies demonstrate the antidepressant activity of the functional NMDA receptor antagonists. In this study, we assessed the effects of two glycine/NMDA receptor ligands, namely L-701,324 (antagonist) and D: -cycloserine (a partial agonist) on the action of antidepressant drugs with different pharmacological profiles in the forced swim test in mice. Swim sessions were conducted by placing mice individually in glass cylinders filled with warmed water for 6 min. The duration of behavioral immobility during the last 4 min of the test was evaluated. The locomotor activity of mice was measured with photoresistor actimeters. L-701,324 and D: -cycloserine given with reboxetine (administered in subeffective doses) did not change the behavior of animals in the forced swim test. A potentiating effect was seen when both tested glycine site ligands were given concomitantly with imipramine or fluoxetine in this test. The lesion of noradrenaline nerve terminals produced by DSP-4 neither altered the baseline activity nor influenced the antidepressant-like action of L-701,324 or D: -cycloserine. The depletion of serotonin by p-CPA did not alter baseline activity in the forced swim test. However, it completely antagonized the antidepressant-like action produced by L-701,324 and D: -cycloserine. Moreover, the antidepressant-like effects of imipramine, fluoxetine and reboxetine were abolished by D: -serine, a full agonist of glycine/NMDA receptors. The present study demonstrates that glycine/NMDA receptor functional antagonists enhance the antidepressant-like action of serotonin, but not noradrenaline-based antidepressants and such their activity seems to depend on serotonin rather than noradrenaline pathway.


Effect of influent feeding pattern on municipal tailwater treatment during a sulfur-based denitrification constructed wetland.
This work studied three parallel pilot-scale constructed wetlands based on sulfur-based autotrophic denitrification (SAD-CWs) with horizontal, vertical-horizontal and integrated vertical inflow for nitrogen removal of municipal tailwater. SAD system played the predominant role for nitrate removal and the integrated vertical inflow pattern was the most efficient pattern with 96.1% NO3--N and 44.3% total phosphorus (TP) removal efficiency, respectively, at the condition of 3.5 h hydraulic retention time (HRT) and 18.5-23.5 °C. Although no great and serious change for microbial community structure was observed among these systems, the diversity in term of abundance of microbes and certain function species was observed. Proteobacteria, Ignavibacterae and Chloroflexi were the dominant phyla and accounted for over 59.1%, 7.5%, and 6.0% in SAD-CWs, respectively. Moreover, the richness and diversity of denitrifies in SAD-CWs with integrated vertical inflow were both higher than that in the other two reactors, especially sulfur autotrophic denitrifying bacteria.


Neuro-imaging and positron emission tomography of congenital homonymous hemianopsia.
Congenital homonymous hemianopsia is an uncommon asymptomatic visual field defect discovered typically in young adult life that is caused by a diverse group of insults to the retrochiasmal afferent visual system occurring prenatally, at birth, or during early childhood. We treated eight patients with congenital homonymous hemianopsia; seven with damage involving the optic radiations and one with an abnormality of the optic tract. We performed positron emission tomography using 18F-fluoro-2-deoxyglucose on two patients with dense homonymous hemianopsias, lesions of the contralateral optic radiations, and largely intact occipital cortex. These studies showed minimal abnormalities in resting visual cortex glucose metabolism of the affected visual cortex.


The Use of Wearable Technology to Objectively Measure Sleep Quality and Physical Activity Among Pregnant Women in Urban Lima, Peru: A Pilot Feasibility Study.
Sleep quality and physical activity can affect the mental and physical health of pregnant women and their babies in utero. We investigated the feasibility of objectively assessing sleep quality and physical activity among resource-constrained, pregnant women in urban Lima, Peru. Twenty pregnant women were asked to complete written sleep logs and wear ActiSleep, a wristwatch-like device that records sleep quality (consecutive minutes of uninterrupted sleep) and physical activity (steps), for seven consecutive days. Sociodemographic data and pregnancy characteristics were also collected. Of twenty women, 13 (65%) had sufficient data collected for analysis. The mean age of study participants was 26.3 years (SD = 3.9), with a mean sleep duration of 6.9 h (SD = 1.4). The median time for sleep onset was 21:15. The mean time for sleep latency was 17.3 min; and wake after sleep onset was 116 min. The mean number of awakenings was 20.4 (SD = 6.7); and sleep efficiency was 77.9%. For physical activity, participants averaged of 6,029 steps per day (SD = 3,087). Objective assessment of sleep quality and physical activity among pregnant women in a resource-constrained setting was promising, despite modest data collection completeness. Wearable technology could be used in health interventions to improve sleep quality and physical activity among this population.


PET imaging of HSV1-tk mutants with acquired specificity toward pyrimidine- and acycloguanosine-based radiotracers.
The aim of this study was to create an alternative mutant of the herpes simplex virus type 1 thymidine kinase (HSV1-tk) reporter gene with reduced phosphorylation capacity for acycloguanosine derivatives, but not pyrimidine-based compounds that will allow for successful PET imaging. A new mutant of HSV1-tk reporter gene, suitable for PET imaging using pyrimidine-based radiotracers, was developed. The HSV1-tk mutant contains an arginine-to-glutamine substitution at position 176 (HSV1-R176Qtk) of the nucleoside binding region of the enzyme. The mutant-gene product showed favorable enzymatic characteristics toward pyrimidine-based nucleosides, while exhibiting reduced activity with acycloguanosine derivatives. In order to enhance HSV1-R176Qtk reporter activity with pyrimidine-based radiotracers, we introduced the R176Q substitution into the more active HSV1-sr39tk mutant. U87 human glioma cells transduced with the HSV1-R176Qsr39tk double mutant reporter gene showed high (3)H-FEAU pyrimidine nucleoside and low (3)H-penciclovir acycloguanosine analog uptake in vitro. PET imaging also demonstrated high (18)F-FEAU and low (18)F-FHBG accumulation in HSV1-R176Qsr39tk+ xenografts. The feasibility of imaging two independent nucleoside-specific HSV1-tk mutants in the same animal with PET was demonstrated. Two opposite xenografts expressing the HSV1-R176Qsr39tk reporter gene and the previously described acycloguanosine-specific mutant of HSV1-tk, HSV1-A167Ysr39tk reporter gene, were imaged using a short-lived pyrimidine-based (18)F-FEAU and an acycloguanosine-based (18)F-FHBG radiotracer, respectively, administered on 2 consecutive days. We conclude that in combination with acycloguanosine-specific HSV1-A167Ysr39tk reporter gene, a HSV1-tk mutant containing the R176Q substitution could be used for PET imaging of two different cell populations or concurrent molecular biological processes in the same living subject.


Fecal pyruvate kinase: a potential new marker for intestinal inflammation in children with inflammatory bowel disease.
Inflammatory bowel disease (IBD) in children creates diagnostic and clinical challenges. Clinical data, endoscopic appearance and the histopathological assessment of biopsies are essential for diagnosis. However, new methods are required for non-invasive follow-up. Recently, we demonstrated that the dimeric isoform of pyruvate kinase (PK) detected in stool might serve as a potential non-invasive screening tool in inflamed pouch mucosa. The aim of this study was to investigate whether this test could be used to detect intestinal inflammation in pediatric IBD patients. Fecal PK immunoreactivity was assessed in 75 patients with proven ulcerative colitis (UC) and 32 with Crohn's disease (CD). Pediatric Crohn Disease Activity Index (PCDAI) and Truelove-Witts scores were determined in CD and UC patients, respectively. Thirty-five healthy subjects (HS) served as a control group. Increased PK levels were documented in 94.1% and 100% active CD patients with a cut-off level of 5 U/g and a cut-off level of 4 U/g, respectively, and in 94.3% of active UC patients regardless of cut-off level. Enzyme immunoreactivity was significantly higher in all IBD patients than in HS. Abnormal PK results were documented in 71.7% of all IBD patients (65.3% and 84.4 for UC and CD patients, respectively). Enzyme levels in UC remission were significantly lower than in the active phase. Enzyme immunoreactivity significantly correlated to both scoring systems. The measurement of stool PK could be a potentially useful marker of IBD activity in children. However, its clinical value demands further studies for comparison with other tests.


Agricultural land-use change and rotation system exert considerable influences on the soil antibiotic resistome in Lake Tai Basin.
In this study, we use high-throughput quantitative polymerase chain reaction approaches to comprehensively assess the effects of agricultural land-use change on the antibiotic resistome of agricultural runoffs after rainfalls in Lake Tai Basin. For the first time in this region, our findings show that orchard runoffs harbored more diverse and abundant antibiotic resistance genes (ARGs) than traditional cropland runoffs. Network analysis demonstrated that orchard runoffs possessed a strong ability for ARG dissemination via horizontal gene transfer. These results suggest that residents might be exposed to a higher public health threat than before. Moreover, the present study confirmed that the rice-wheat rotation system plays a key role in regulating the soil antibiotic resistome profile. Using 16S rRNA high-throughput sequencing technology, this study clarified the relationships between the antibiotic resistome and soil microbiome composition. Finally, we discuss the key environmental factors driving changes in the soil antibiotic resistome. In summary, this study gives insight into the dissemination of environmental ARGs to the people living in the Lake Tai Basin.


Effect of physical and chemical properties of vanadium slag from stone coal on the form of vanadium.
Vanadium mining and smelting activities were increasing extensively and causing serious vanadium pollution in soil around the mining area. Different existing forms of vanadium had different biological effects and the exchangeable state had been recognized as a severe threat to biodiversity and ecosystem functioning. At present, the research on vanadium morphology had not received much attention. In this study, the area that we researched had been severely polluted with vanadium due to mining and smelting activities. The changes in the morphology of vanadium in soil were studied by adjusting the organic matter content, clay mineral content, pH value, and Eh value. The results showed that at pH 8 and for 1% of humic acid added, the exchangeable fraction of vanadium in the slag was 10% and 9%, respectively, which was 5% and 6% lower than the control group. The addition of kaolin and the redox change had little effect on the exchangeable fraction of vanadium, with a change of only about 2%. To control the soil pollution caused by slag and to repair its ecological characteristics, kaolin and humic acid were used for the repair test. The results showed that after 1% humic acid mixed with 8% kaolin was added in soil, the germination rate of ryegrass reached 95% and grew flourishingly which is significantly better than other treatment groups. Our research can provide a reference for future vanadium pollution control, especially in the morphology of vanadium research.


[Cardiovascular disorder syndrome in sepsis].
The authors have observed 126 patients with sepsis and made a conclusion that all of them had cardio-vascular insufficiency. The use of hyperbaric oxygenation resulted in better activity of the cardio-vascular system.


Corona, Climate Change, and Evolved Human Behavior.
Most scientists agree that we have to restrict climate change, but there is much frustration that we are failing. The Corona Crisis exemplifies how human behavior is constrained by its evolution, cognition, and resource availability, explaining why we do not act to avoid climate change for the benefit of future generations.


Hand Function in 8- to 12-Year-Old Children with Bilateral Cerebral Palsy and Interpretability of the Both Hands Assessment.
To describe bimanual performance in a sample of Australian children with bilateral cerebral palsy (CP) and, examine the qualitative meaning (or interpretability) of scores on the Both Hands Assessment (BoHA). Children with bilateral CP aged 8-12 years (n = 54) classified Manual Ability Classification System (MACS) level I = 20, II = 18, III = 16 were examined using the BoHA. Bimanual performance was significantly different across MACS levels I-III (p < 0.001). Mean (95%CI) BoHA-unit for each MACS level were I = 85 (81-89), II = 72 (68-76) and III = 53 (49-56). Children with asymmetrical hand use (≥ 20% difference between upper limbs, n = 10) were classified MACS levels II and III and had a mean (95%CI) BoHA-unit of 56 (51-62). Children with symmetrical hand use were classified in MACS level I-III and had a mean (95%CI) BoHA-unit of 74 (70-79). The BoHA quantified observations of bimanual performance for children with bilateral CP, differentiated between MACS levels I-III and provided clinically meaningful information. The BoHA may facilitate tailoring of upper limb intervention. Future research is recommended to examine inter-rater and intra-rater reliability and responsiveness of the BoHA, as well as longitudinal studies of bimanual hand skill development in children with bilateral CP.


[Susac syndrome: an interdisciplinary challenge].
Susac syndrome, named after John Susac, the first to describe this condition, is characterized by the clinical triad of encephalopathy, branch retinal artery occlusion, and sensorineural hearing loss. Although certainly a rare disease, Susac syndrome needs to be considered in the differential diagnosis of a broad variety of diseases. The pathogenesis is not yet clear. Autoimmune processes leading to damage and inflammation-related occlusion of the microvessels in brain, retina, and inner ear are thought to play a causal role. The diagnosis is based primarily on the clinical presentation, the documentation of branch retinal artery occlusion by fluorescence angiography, and characteristic findings on cerebral MRI. Usually, immunosuppressive therapy is required, though controlled therapy trials are missing so far. The intention of this review article is to raise awareness of this disease among neurologists, psychiatrists, ophthalmologists, and ENT specialists as a high number of unreported cases probably exists. Accordingly, the focus is on the clinical presentation and the diagnostic approach.


A model integrating multiple processes of synchronization and coherence for information instantiation within a cortical area.
What is the form of dynamic, e.g., sensory, information in the mammalian cortex? Information in the cortex is modeled as a coherence map of a mixed chimera state of synchronous, phasic, and disordered minicolumns. The theoretical model is built on neurophysiological evidence. Complex spatiotemporal information is instantiated through a system of interacting biological processes that generate a synchronized cortical area, a coherent aperture. Minicolumn elements are grouped in macrocolumns in an array analogous to a phased-array radar, modeled as an aperture, a "hole through which radiant energy flows." Coherence maps in a cortical area transform inputs from multiple sources into outputs to multiple targets, while reducing complexity and entropy. Coherent apertures can assume extremely large numbers of different information states as coherence maps, which can be communicated among apertures with corresponding very large bandwidths. The coherent aperture model incorporates considerable reported research, integrating five conceptually and mathematically independent processes: 1) a damped Kuramoto network model, 2) a pumped area field potential, 3) the gating of nearly coincident spikes, 4) the coherence of activity across cortical lamina, and 5) complex information formed through functions in macrocolumns. Biological processes and their interactions are described in equations and a functional circuit such that the mathematical pieces can be assembled the same way the neurophysiological ones are. The model can be conceptually convolved over the specifics of local cortical areas within and across species. A coherent aperture becomes a node in a graph of cortical areas with a corresponding distribution of information.


Dental pulp stem cell migration.
The purpose of this in vitro study was to investigate the migration of dental pulp stem cells (DPSCs) in response to chemotactants and extracellular matrix proteins (EMPs). This DPSC signaling information is needed to help understand tooth regeneration after injury and to develop some future regenerative endodontic therapies. DPSCs were released by trypsinization and plated on transwell filters. The chemotactants were recombinant sphingosine-1-phosphate (S1P), fibroblast growth factor (FGF), epidermal growth factor (EGF), or transforming growth factor beta-1 (TGF-β1), and the EMPs were collagen-1, collagen-IV, laminin, and fibronectin. Data were analyzed by using analysis of variance (ANOVA) statistical tests for cell migration. S1P induced more vigorous DPSC migration in comparison with the other TGF- β1, FGF, or EFG chemotactants (ANOVA, P < .05). Laminin induced more vigorous DPSC migration in comparison with the other EMPs (ANOVA, P < .05). The EMPs, particularly laminin, and chemotactants, particularly S1P and TGF-β1, were found to be important promoters of DPSC migration. The interplay between the EMPs, blood lipid, serum, and chemotactants suggests that the migration of DPSC is highly regulated. Specific chemotactants and EMPs might mediate the process of pulp-dentin regeneration after tooth injury, and they could be used as part of regenerative endodontic therapy.


Evidence for four-component close-range sex pheromone in the parasitic wasp Glyptapanteles flavicoxis.
Females of the parasitic wasp Glyptapanteles flavicoxis (Hymenoptera: Braconidae) deposit a close-range sex pheromone from their abdominal tip that attracts conspecific males and elicits wing-fanning behavior. In this study, we isolated the pheromone components and determined their role in the males' behavior. In coupled gas chromatographic-electroantennographic detection (GC-EAD) analyses of the females' body extract, four components (below GC detection) elicited strong responses from male antennae. Monitored by GC-EAD, the components were separated by flash silica gel and high-performance liquid chromatography. Y-tube olfactometer experiments with one or more components revealed that all are necessary to elicit short-range attraction and wing-fanning responses by males. These components remained below detection threshold of the mass spectrometer (approximately 10 pg) even when 4500 female equivalents were analyzed in a single injection, which attests to the potency of the pheromone and the insects' sensitivity to it.


Deposition of contaminant aerosol on human skin.
Over recent years, it has been established that deposition of various types of pollutant aerosols (e.g., radioactive) on human skin can have serious deleterious effects on health. However, only few investigations in the past have been devoted to measurement of deposition velocities on skin of particles of the potentially problematic sizes. An experimental programme has shown the deposition velocities on skin of particles in the ca. 0.5-5 microm AMAD range to be high and generally associated with great variations. A series of investigations have been made to identify some of the factors that lead to this variation. Part of the variation was found to be caused by differences between individuals, whereas another part was found to be related to environmental factors. The identification of major influences on skin contaminant deposition is important in estimating health effects as well as in identifying means for their reduction.


Nuclear magnetic resonance structural studies of membrane proteins in micelles and bilayers.
Nuclear magnetic resonance (NMR) spectroscopy enables determination of membrane protein structures in lipid environments, such as micelles and bilayers. This chapter outlines the steps for membrane-protein structure determination using solution NMR with micelle samples, and solid-state NMR with oriented lipid-bilayer samples. The methods for protein expression and purification, sample preparation, and NMR experiments are described and illustrated with examples from gamma and CHIF, two membrane proteins that function as regulatory subunits of the Na+- and K+-ATPase.


Differential expression of inducible nitric oxide synthase and cytokine mRNA in chicken lines divergent for cutaneous hypersensitivity response.
Phytohemagglutinin (PHA)-induced delayed-type hypersensitivity is an immunocompetent trait considered an indicator of cell-mediated immune or T-cell responses. Divergent selection was performed to generate high and low lines for response to PHA-P. Extreme-responder birds of the F2 generation in each line were used to study possible differences in macrophage activity and the associated functional genes. To evaluate macrophage activity, nitric oxide (NO) was estimated both systemically in serum and in in vitro monocyte culture. Semi-quantitative RT-PCR was used to detect the differential mRNA expression patterns of iNOS and MIP-1beta in monocyte culture, whereas T(H)1 cytokines (IL-2 and IFN-gamma) were studied in peripheral blood mononuclear cells (PBMC) at different time intervals after lipopolysaccharide (LPS) induction. The high line showed strong systemic, as well as in vitro NO production, compared to the low line, upon stimulation with NDV and LPS, similar to early and high iNOS mRNA expression. Following the pattern of iNOS gene expression, an early strong expression of cytokines with powerful iNOS-inducing action, such as IFN-gamma and the chemokine MIP-1beta, was observed in the high line. In contrast, for response to PHA-P, low expression of IL-2 was observed in the high compared to the low line. In conclusion, the study revealed that divergent selection for response to PHA-P resulted in a divergent effect on T(H)1 cell activity, resulting in altered macrophage function in chickens. Selection, based on response to PHA-P, could lead to more resistant birds or birds with an enhanced immune response.


Integrated metabolomics and mechanism to reveal the protective effect of kaempferol on pulmonary arterial hypertension.
Currently, there have been breakthroughs in the study of the underlying mechanisms of pulmonary arterial hypertension, but treatment is still challenging. The aim of this study was explore the effect of kaempferol in PAH and discover the mechanism of this process. First, we assessed the effect of kaempferol in animal models of PAH. The echocardiography and pressure-volume admittance catheter data showed that kaempferol could improve right ventricular function and alleviate the progression of PAH. Moreover, morphometric analysis of the lung vasculature indicated that pulmonary vascular remodeling could be reduced by kaempferol. The expression of autophagy-related proteins and autophagic processes were evaluated by western blotting and mRFP-GFP-LC3 fluorescence microscopy, the results indicated that autophagy played an important role in the process by which kaempferol prevents PAH. Moreover, we performed untargeted and targeted plasma metabolomics analysis on rat models of PAH and used multivariate analysis to reveal multiple pathways and targets. Consequently, we identified 247 plasma biomarkers that were tightly linked to the preventive effects of kaempferol, which were mainly related to amino acid metabolism and arachidonic acid metabolism. In conclusion, kaempferol could effectively alleviate the development of PAH by regulating abnormal autophagy and metabolic disorders, providing a novel perspective on the treatment of PAH.


Rat CNS in experimental chronic serum sickness: integrity of the zonulae occludentes of the choroid plexus epithelium and brain endothelium in experimental chronic serum sickness.
The present study is an extensive systematic immunofluorescence and labelled electron microscopic investigation of chronic rat serum sickness brains. It is specifically directed at discerning effects of chronic serum sickness upon the zonulae occludentes of both the choroid plexus epithelium and the intra-cerebral endothelium. Although the choroid plexus is a known site of complex entrapment in systemic immune complex disorders, comprehensive immune studies of the CNS parenchymal vasculature have not yet been reported in either experimental or spontaneous immune complex disease. We examined by direct immunofluorescence techniques over 70 blocks of experimental chronic serum sickness brains taken from 16 animals with severe glomerular disease and found them to be uniformly negative for the presence of immune deposits. No horseradish peroxidase was seen beyond the restricting zonulae occludentes of either the choroid plexus or the cerebral endothelium in tissue from three serious affected animals. These structural barriers retained their integrity despite the extensive circulation and deposition of complexes and complement induced in the experimental model and the associated release of vasoactive substances.


Identification of mucin degraders of the human gut microbiota.
Mucins are large glycoproteins consisting of approximately 80% of hetero-oligosaccharides. Gut mucin degraders of healthy subjects were investigated, through a culture dependent and independent approach. The faeces of five healthy adults were subjected to three steps of anaerobic enrichment in a medium with sole mucins as carbon and nitrogen sources. The bacterial community was compared before and after the enrichment by 16S rRNA gene profiling. Bacteria capable of fermenting sugars, such as Anaerotruncus, Holdemania, and Enterococcaceae likely took advantage of the carbohydrate chains. Escherichia coli and Enterobacteriaceae, Peptococcales, the Coriobacteriale Eggerthella, and a variety of Clostridia such as Oscillospiraceae, Anaerotruncus, and Lachnoclostridium, significantly increased and likely participated to the degradation of the protein backbone of mucin. The affinity of E. coli and Enterobacteriaceae for mucin may facilitate the access to the gut mucosa, promoting gut barrier damage and triggering systemic inflammatory responses. Only three species of strict anaerobes able to grow on mucin were isolated from the enrichments of five different microbiota: Clostridium disporicum, Clostridium tertium, and Paraclostridium benzoelyticum. The limited number of species isolated confirms that in the gut the degradation of these glycoproteins results from cooperation and cross-feeding among several species exhibiting different metabolic capabilities.


Ultrastructural study of differentiation processes during aggregation of purified sponge archaeocytes.
Archaeocytes from the spongeEphydatia fluviatilis were dissociated and then isolated on Ficoll density gradients. Their aggregation and reconstitution processes were studied by transmission electron microscopy to determine their capabilities for differentiation.Archaeocyte aggregates follow a well defined sequence of differentiation to generate the characteristic structures of a sponge. Pinacoderm is the first structure to be regenerated and appears progressively at the surface of the 12 h aggregates. Pinacocytes which have differentiated in archaeocyte aggregates are identical to native ones except that the nucleolus remains in most cells. The choanocytes appear only after 24 h by a two step process. First, small cells (choanoblasts) are formed from archaeocytes by mitosis. These cells then transform into fully differentiated choanocytes possessing collars and flagella. The early choanocyte chambers are small, irregular and randomly dispersed in the aggregates. Finally, collencytes and sclerocytes begin to appear just before the aggregates spread on the substrate.The differentiation of a suspension of pure archaeocytes is a unique model system to study sponge cell differentiation and has allowed us to demonstrate that archaeocytes isolated from developed sponges maintain the capacity to differentiate even though this capacity is not usually expressed.


Long-Term Outcomes of the Maze Procedure Combined with Endoatriectomy for Patients with Calcified Left Atria.
Left atrial calcification (LAC) is found in long-lasting rheumatic valvular disease and is almost always accompanied by atrial fibrillation (AF). In the presence of LAC, endoatriectomy is required when performing the maze procedure. However, the technical feasibility of endoatriectomy and the long-term outcomes of the maze procedure in patients with LAC are uncertain. The medical records of 18 consecutive patients who underwent combined endoatriectomy and maze procedure were analyzed retrospectively. Accompanying operations were mitral valve replacement (n = 16) and commissurotomy (n = 2). There was 1 operative death from sepsis following mediastinitis. When patients were divided into "broad" (n = 11) and "limited" (n = 7) groups, with the extent of LAC either greater or less than half of the left atrium, respectively, there was no intergroup difference in postoperative complications. During follow-up (median, 11.4 years), AF recurred in 11 patients. At the last follow-up visits, electrocardiography revealed significantly fewer patients in the broad group maintaining sinus rhythm (1/11 vs 4/6, P = 0.03). The 10-year AF recurrence-free survival rates were 13.9% and 66.7% in the broad and limited groups, respectively (P = 0.01). The maze procedure combined with endoatriectomy seems technically feasible with acceptable long-term rhythm outcomes if the LAC extent is limited.


Love me Tinder: Body image and psychosocial functioning among men and women.
Based on objectification theory, we examined the main effects of Tinder use, and its interaction with gender, in relation to men's and women's body image concerns, internalization processes, and self-esteem. Tinder users (men=31; women=69) and non-users (men=203; women=844) anonymously completed measures via an online survey. Through a series of ANCOVAs, with BMI and age as covariates, Tinder users, regardless of gender, reported significantly lower levels of satisfaction with face and body and higher levels of internalization, appearance comparisons, and body shame and surveillance than non-users. For self-esteem, male Tinder users scored significantly lower than either male or female non-users. Our results suggest that Tinder represents a contemporary medium for appearance pressures and its use is associated with a variety of negative perceptions about body and self and with increases in individuals' likelihood to internalize appearance ideals and make comparisons to others.


Human clinical trials of plasmid DNA vaccines.
This article gives an overview of DNA vaccines with specific emphasis on the development of DNA vaccines for clinical trials and an overview of those trials. It describes the preclinical research that demonstrated the efficacy of DNA vaccines as well as an explication of the immunologic mechanisms of action. These include the induction of cognate immune responses, such as the generation of cytolytic T lymphocytes (CTL) as well as the effect of the plasmid DNA upon the innate immune system. Specific issues related to the development of DNA as a product candidate are then discussed, including the manufacture of plasmid, the qualification of the plasmid DNA product, and the safety testing necessary for initiating clinical trials. Various human clinical trials for infectious diseases and cancer have been initiated or completed, and an overview of these trials is given. Finally, because the early clinical trials have shown less than optimal immunogenicity, methods to increase the potency of the vaccines are described.


Other manifestations of mixed connective tissue disease.
This article describes other manifestations of mixed connective tissue disease (MCTD). We focus on inflammatory arthritis, the gastrointestinal tract (the esophagus in particular), the kidney, skin, and hematologic changes such as thrombocytopenia. Due to the complexity of potential organ involvements in MCTD, vigilance with appropriate diagnosis and treatment is warranted.


Recombinant gp160 vaccination schedule and MHC HLA type as factors influencing cellular responses to HIV-1 envelope glycoprotein. NIAID AIDS Vaccine Clinical Trials Network.
This study assessed (1) T-cell responses following vaccination of HIV-1 negative volunteers with HIV-1LAI recombinant gp160 (rgp160) vaccine, and (2) effects of vaccine injection schedule and MHC HLA type on those responses. In one trial, volunteers received rgp160 in two accelerated schedules (Groups 1 and 2). In the other trial, volunteers received rgp160 vaccine at 0, 1, 6, and 12 months and some received vaccine at 18 months (Groups 3 and 4). Peripheral blood mononuclear cells (PBMC) from vaccinees in Groups 3 and 4 were stimulated by more peptides representing envelope glycoprotein T-cell epitopes than Groups 1 and 2 (p = 0.01). Transient anti-envelope glycoprotein CTL activity was more frequently detectable in Groups 1 and 2 (p = 0.03). MHC HLA-Cw7, DR1 and DQw1 alleles appeared to be associated with PBMC responses to envelope glycoprotein.


Many steps can be taken to enhance recovery after thoracic surgery.
Our letter to the editor comments on issues raised in the May 14, 2020, article by Budacan et al. addressing the development of enhanced recovery after thoracic surgery. In the United Kingdom and Ireland, a nationwide survey identified issues. Here, we expand on the authors' findings.


In vivo absolute quantification of carnosine in the vastus lateralis muscle with 1H MRS using a surface coil and water as internal reference.
To standardize a method for 1H MRS intramuscular absolute quantification of carnosine in the thigh, using a surface coil and water as internal reference. Carnosine spectra were acquired in phantoms (5, 10, and 15 mM) as well as in the right gastrocnemius medialis (GM) and right vastus lateralis (VLM) muscles of young team sports athletes, using volume (VC) and surface (SC) coils on a 3 T scanner, with the same receiver gain. Water spectra were used as internal reference for the absolute quantification of carnosine. Phantom's experiments showed a maximum error of 7%, highlighting the validity of the measurements in the study setup. The carnosine concentrations (mmol/kg ww, mean ± SD) measured in the GM were 6.8 ± 2.2 with the VC (CcarVC) and 10.2 ± 3.0 with the SC (CcarSC) (P = 0.013; n = 9). Therefore, a correction was applied to these measurements (CcarVC = 0.6582*CcarSC), to make coils performance comparable (6.8 ± 2.2 for VC and 6.7 ± 2.0 for SC, P = 0.97). After that, only the SC was used to quantify carnosine in the VLM, where a concentration of 5.4 ± 1.5 (n = 30) was found, with significant differences between men (6.2 ± 1.3; n = 15) and women (4.6 ± 1.2; n = 15). The error in quantitation was 5.3-5.5% with both coils. The method using the SC and water as internal reference can be used to quantify carnosine in voluminous muscles and regions of the body in humans, where the VC is not suitable, such as the VLM.


Multimodal Imaging in an Unusual Cluster of Multiple Evanescent White Dot Syndrome.
To describe an unusual cluster of multiple evanescent white dot syndrome (MEWDS) encountered within a 3-month period. This retrospective observation study is comprised of seven patients who presented with MEWDS in a 3-month period in central Israel. Data were collected from patients' medical records on clinical, multimodal imaging, and viral serology findings. Six women and one man of mean age 31.5 ± 7.2 years. Three reported a precedent viral infection. All had unilateral decreased vision. Funduscopy revealed foveal granularity. Hyperfluorescent spots on blue autofluorescence (BAF), hypofluorescent spots on indocyanine green angiography, dark lesions on infrared photos, and ellipsoid zone irregularities on spectral domain optical coherence tomography (SD-OCT). Resolution of the spots on BAF correlated with anatomic (SD-OCT) and visual recovery. OCT angiography performed following the convalescence stage demonstrated intact retinal and choroidal flow. Serologic findings were inconclusive. We report a unique cluster of MEWDS patients presented in a short period of time. SD-OCT findings of ellipsoid zone disruption in combination with other multimodal imaging modalities are outlined meticulously. Recognizing these imaging features along with high index of clinical suspicion is important for the diagnosis of MEWDS. Serologic testing might be considered in future patients.


Current management of limited-stage SCLC and CONVERT trial impact: Results of the EORTC Lung Cancer Group survey.
The CONVERT trial showed that twice-daily (BD) concurrent chemoradiotherapy should continue to be considered the standard of care in localised LS-SCLC. A survey was conducted to assess the impact of the CONVERT trial in clinical practice and to identify any relevant research questions for future trials in this setting. An EORTC Group online survey of LS-SCLC practice was distributed to the EORTC LCG and to members of several European thoracic oncology societies between April and December 2018. 198 responses were analysed. The majority of respondents (88%, n = 174) were aware of the CONVERT trial. Radiation oncologists comprised 56% of all respondents. Once-daily (OD) radiotherapy is still the most commonly used regimen, however the use of concurrent BD radiotherapy increased after the publication of CONVERT (n = 59/186, 32% prior to and n = 78/187, 42% after the publication, p = 0.053). The main reasons for not implementing BD after the CONVERT publication were logistical issues (n = 88, 44%), inconvenience for patients (n = 56, 28%), and the absence of a statistical survival difference between the two arms in CONVERT (n = 38, 19%). Brain MRI was used by 28% during staging but more than half (60%) of the respondents did not routinely image the brain during follow-up. The main research questions of interest in LS-SCLC were 1) integrating novel targeted therapies-immunotherapies (n = 160, 81%), 2) PCI (+/- hippocampal sparing) vs. MRI surveillance (n = 140, 71%) and, 3) biomarker driven trials (n = 92, 46%). Once daily radiotherapy (60-66 Gy in 30-33 fractions) remains the most prescribed radiotherapy fractionation, despite the findings suggested by the CONVERT trial.


Comparing the Beck Depression Inventory-II (BDI-II) and Patient Health Questionnaire (PHQ-9) depression measures in an integrated mood disorders practice.
Patient self-assessment instruments are useful in screening, tracking, and documenting the course of depressive symptoms with minimal clinician time. Two popular instruments are the Beck Depression Inventory (BDI-II) and the Patient Health Questionnaire (PHQ-9). We compared the performance of these two instruments in a mood disorders setting. A retrospective study of 625 patients who completed a PHQ-9 and BDI-II as part of routine clinical care (1) during initial outpatient evaluation between 2008 and 2009, and (2) on admission to an inpatient mood disorders unit between 2006 and 2009. Pearson correlation coefficients for total PHQ-9 and BDI-II scores were calculated for all patients, the outpatients, and the inpatients. Overall r=0.77, indicating strong correlation, more in the outpatients (n=287, r=0.81) than the inpatients (n=338, r=0.67). Mean PHQ-9 and BDI-II scores for the outpatients were 15.1 (SD 7.4) and 27.8 (SD 14.1) corresponding to "moderately severe" and "moderate" categories respectively; for inpatients, 18.9 (SD 5.7) and 33.8 (SD 11.5) corresponding to "moderately severe" and "severe." Retrospective design and no monitoring of which instrument was completed first in case that influenced patient response to the second instrument PHQ-9 and BDI-II scores, as continuous but not categorical variables, in a mood disorders subspeciality setting are closely correlated and essentially interchangeable. There are practical applications to our findings, as the PHQ-9 is shorter and free.


Effect of sex and age on serum aldosterone and thyroid hormones in the laboratory rat.
Serum levels of aldosterone, tri-iodo thyronine (T3) and thyroxine (T4) were measured in male and female rats aged 3 months, 12 months, and 18 months. Female rats were found to have higher aldosterone and T3 levels, and lower T4 level than the male. No age-related change was observed in serum aldosterone in either sex. In contrast, serum T4 were found to decrease with age in both sexes while serum T3 showed an age-associated diminution in the male only. Serum testosterone was also measured in the male rats and was found to decline with age.


Probing the dioxygen route in Melanocarpus albomyces laccase with pressurized xenon gas.
Laccases catalyze the oxidation of phenolic substrates and the concominant reduction of dioxygen to water. We used xenon as an oxygen probe in search of routes for the entry of dioxygen into the catalytic center. Two xenon-pressurized crystal structures of recombinant Melanocarpus albomyces laccase were determined, showing three hydrophobic Xe-binding sites located in domain C. The analysis of hydrophobic cavities in other laccase structures further suggested the preference of domain C for binding of hydrophobic species such as dioxygen, thus suggesting that the hydrophobic core of domain C could function as a channel through which dioxygen can enter the trinuclear copper center.


Potassium channels, memory T cells, and multiple sclerosis.
Multiple sclerosis is a chronic inflammatory autoimmune disease of the central nervous system characterized by demyelination and axonal damage that result in disabling neurological deficits. Here the authors explain the rationale for the use of inhibitors of the Kv1.3 K+ channel in immune cells as a therapy for multiple sclerosis and other autoimmune disorders.


[The problem of residual cavity and local recurrences in surgical treatment of hepatic echinococcosis].
Results of examination and treatment of 326 patients, suffering hepatic echinococcosis, in whom 449 surgical interventions were performed, were analyzed. In 123 patients the disease recurrence was diagnosed. In 80 (17.8%) patients laparoscopic access was applied, in 261 (58.1%)--laparotomic, in 108 (24.1%)--the puncture treatment of parasitic cysts under ultrasonographic investigation control. There was established, that the residual cavity walls after echinococcectomy, if the condition of their chemical and thermal processing do not constitute a danger for the disease recurrence occurrence, but the residual cavity suppuration or formation of durably persisting residual cavities in liver is possible. So the optimal operative intervention must be echinococcectomy with partial resection of fibrose capsule with the thinned hepatic parenchyma.


Branched-chain amino acids protect the liver from cirrhotic injury via suppression of activation of lipopolysaccharide-binding protein, toll-like receptor 4, and signal transducer and activator of transcription 3, as well as Enterococcus faecalis translocation.
Branched-chain amino acids (BCAAs) are used as nutritional support and for improving prognosis in liver cirrhosis. Here we investigate the molecular mechanisms of BCAA treatment and liver damage focused on pathways related to lipopolysaccharide-binding protein (LBP). Serum LBP levels were measured in cirrhotic patients and in cirrhotic rats treated with BCAA to examine the correlation between liver function and survival. In cirrhotic rats, liver damage, Enterococcus faecalis translocation, serum capsular polysaccharide, and intestinal tight junction levels were assessed. Damaged HepG2 cells were cultured with BCAA-supplemented, BCAA-deficient, or control amino acid medium, followed by examination of LBP expression. Serum LBP levels were significantly increased in deceased patients individuals with liver cirrhosis. The survival rate in patients with lower serum LBP (<3.48 μg/mL) was significantly improved. In BCAA-treated rat liver samples, protein expression of LBP, toll-like receptor 4 (TLR4), and phosphorylated signal transduction and activator of transcription 3 (STAT3) were significantly reduced. Also in BCAA-treated rats, intestinal zonula occludens gene expression was increased, whereas hepatic translocation of E. faecalis and serum capsular polysaccharide levels were reduced. In damaged HepG2 cells, lipopolysaccharide-induced elevation of LBP expression was rapidly and strongly repressed in BCAA-enriched medium. Serum LBP level is a prognostic biomarker in liver cirrhosis. BCAA treatment reduced translocation of E. faecalis through intestinal tight junction recovery and reduced LBP expression in the liver, which repressed activation of LBP, toll-like receptor 4, and signal transduction and activator of transcription 3. Our findings suggest that BCAA supplementation protects the liver from damage via multiple pathways.


Homozygous PLA2G6 (PARK 14) gene mutation associated neuropsychiatric phenotypes from southern India.
PLA2G6 gene associated neurodegenerative disorders resulting from homozygous c. 2222G > A (p.Arg741Gln) mutation were detected in two cases having variable neuropsychiatric phenotypic and imaging findings. Exome analysis helped identification of rare alleles, reinforcing ethnographic antecedents to geographical clustering of rare mutations and, essential to understanding biology of neurodegenerative disorders.


The prevalence of ADHD-like symptoms in a community sample.
The aim of the present study was to explore the prevalence of ADHD symptoms in a community sample of children in the United Kingdom. There were 964 ten year olds (55% boys; M = 10.4 years; SD = 0.6) from mainstream schools in the United Kingdom in this study. The ADHD Rating Scale-IV, which provides ratings on the frequency of ADHD symptoms drawn from DSM-IV criteria, was administered to class teachers. The findings indicated an overall 8% prevalence rate, with the majority of children identified as the Hyperactive/Impulsive subtype (5%). Almost half of these children were receiving additional support in the classroom as a result of learning difficulties. It is therefore of value for educators to be able to conduct early screening to support these children before problems worsen.


Long-term cimetidine in duodenal ulcer disease.
Forty patients with chronic duodenal ulcer who had healed endoscopically with a 6-week course of cimetidine were randomized double blind to 1 year of either placebo or cimetidine tablets 400 mg bid (20 patients in each group). Patients were seen at monthly intervals, and endoscopy was performed at clinical relapse or on completion of 1 year. One of 20 patients on active cimetidine relapsed clinically and endoscopically at 3 months; 16 of 20 patients on placebo relapsed clinically and endoscopically within 9 months, the majority within 3 months, and 2 were shown to have asymptomatic chronic ulcers at routine 12-month endoscopy. None of the 19 patients on active cimetidine routinely endoscoped at 12 months showed evidence of ulceration. This study confirms a high relapse rate when short-term cimetidine is ceased and indicates that maintenance treatment with cimetidine prevents relapse.


Antimicrobial defenses of table eggs: Importance of antibacterial proteins in egg white as a function of hen age in an extended production cycle.
The importance of egg natural defences to prevent bacterial contamination and their relation with hen age in extended production cycles were evaluated. Egg-white from eggs of different hen age groups (up 100-weeks-old) and lines (Hy-Line white and brown) were inoculated with Gram-positive Staphylococcus aureus or Gram-negative Salmonella Typhimurium, ranging from 103-106 CFU/mL. Our results show that concentrations of egg-white lysozyme and, particularly, ovotransferrin are important to modulate bacterial survival in a dose-dependent matter. Depending on protein concentration, their effect ranges from bactericidal to bacteriostatic, with a threshold for bacterial contamination that depends also on hen age and line. The concentrations of lysozyme and ovotransferrin increased with hen age (up to 2 and 22 w/w% of total protein, respectively), and eggs laid by older hens exhibited the greatest potential to prevent the growth of the highest Salmonella inoculum (106 CFU/mL). Salmonella-penetration experiments demonstrated that non-contaminated eggs display significantly higher concentrations of antimicrobial proteins. However, eggs from older hens needed a higher concentration of these proteins (>20% ovotransferrin) to prevent bacterial contamination, showing that antimicrobial protein concentrations in egg-whites was not the only factor influencing bacterial contamination. Finally, this study demonstrated that egg-white of eggs produced by old hens are less prone to contamination by Salmonella.


Cirrhotic Patients with Bacterial Infection and Negative Cultures Have a More Advanced Disease and an Increased Short-Term Mortality Rate.
The negative clinical impact of bacterial infections (BI) in patients with cirrhosis is well documented. In cirrhotic patients, failure to isolate the pathogen is a frequent event, occurring in 30-40% of cases. The aim of this study was to compare the clinical characteristics, early (30-day) and short-term (90-day) mortality rates, in a cohort of cirrhotic patients with BI, between those with positive (C-pos) and those with negative (C-neg) microbiological cultures. We retrospectively enrolled 279 consecutive hospitalized cirrhotic patients with BI. Survival and predictors of 30-day and 90-day mortality were assessed by Kaplan-Meier curves and logistic regression analysis, respectively. Cultures tested negative in 108/279 (38.7%) patients. C-neg patients were more frequently males (p = 0.035), had higher Child-Pugh-Turcotte (CPT; p = 0.007) and model for end-stage liver disease-sodium (MELD-Na; p = 0.043) scores, and had more frequently decompensated liver disease (p = 0.04). Mortality rate was higher in C-neg than in C-pos patients, both at 30 days (22.2% versus 11.7%, p = 0.024) and 90 days (46.3% versus 33.3%, p = 0.030). MELD-Na score and non-selective beta-blockers (NSBBs) were independent risk factors for 30-day and 90-day mortality. In particular, the use of NSBBs was independently associated with a lower 30-day and 90-day mortality risk (OR 0.41, CI95% 0.17-0.94, p = 0.040; and OR 0.43, CI95% 0.25-0.75, p = 0.003, respectively). Cirrhotic patients with BI and negative microbiological cultures have significantly higher mortality compared to those with positive cultures. Early mortality and short-term mortality are mainly influenced by the underlying severity of liver disease. In this contest, therapy with NSBBs has a positive impact on short-term survival.


Health-related quality of life assessment in prospective trials of systemic cytotoxic chemotherapy for metastatic castration-resistant prostate cancer: which instrument we need?
Knowledge about psychological health of men with prostate cancer is still limited. HRQoL assessment adds value in symptom management by allowing a broader understanding of the impact of symptom management beyond the targeted symptom, on functioning, and on overall QoL. In this paper, the results of the commonly used HRQoL questionnaires in phase III randomized clinical trials of chemotherapy in metastatic castration-resistant prostate cancer has been discussed. An overview about symptom burden, treatments and HRQoL domains, a description of available HRQoL instruments used for patients with metastatic castration-resistant prostate cancer were reported. Finally, the characteristics of most commonly used HRQoL instruments were identified and compared. To provide better empirical justification for the selection of HRQoL instruments, head-to-head comparisons of them within the same studies are needed. Estimating a minimal important difference could be significant when interpreting trial results. The impact of HRQoL scores in clinical practice remains unclear; poor communication of clinical significance of the results and limited training of clinicians are the most important barriers to a widespread use of HRQoL questionnaires.


Helium-Surface Interaction and Electronic Corrugation of Bi2Se3(111).
We present a study of the atom-surface interaction potential for the He-Bi2Se3(111) system. Using selective adsorption resonances, we are able to obtain the complete experimental band structure of atoms in the corrugated surface potential of the topological insulator Bi2Se3. He atom scattering spectra show several selective adsorption resonance features that are analyzed, starting with the free-atom approximation and a laterally averaged atom-surface interaction potential. Based on quantum mechanical calculations of the He-surface scattering intensities and resonance processes, we are then considering the three-dimensional atom-surface interaction potential, which is further refined to reproduce the experimental data. Following this analysis, the He-Bi2Se3(111) interaction potential is best represented by a corrugated Morse potential with a well depth of D = (6.54 ± 0.05) meV, a stiffness of κ = (0.58 ± 0.02) Å-1, and a surface electronic corrugation of (5.8 ± 0.2)% of the lattice constant. The experimental data may also be used as a challenging benchmark system to analyze the suitability of several van der Waals approaches: the He-Bi2Se3(111) interaction captures the fundamentals of weak adsorption systems where the binding is governed by long-range electronic correlations.


Oxolinic acid--trimethoprim combination: effects on DNA synthesis and on viability of Escherichia coli.
The synergistic bactericidal effect of the oxolinic acid-trimethoprim combination has been confirmed on an exponentially growing strain of Escherichia coli, and is paralleled by a synergistic inhibition of 14C-uracil incorporation into the bacterial DNA. These data provide experimental support for the postulate that the synergistic effect of quinolone-trimethoprim combinations, observed both in vitro and in vivo, is due to a complementary blockade of bacterial DNA synthesis.


Molecular modeling of CD28 and three-dimensional analysis of residue conservation in the CD28/CD152 family.
CD28/CD152-CD80/CD86 receptor-ligand interactions result in costimulatory signals critical for optimal T cell activation. CD28/CD152 and CD80/CD86 are members of the immunoglobulin superfamily (IgSF). Despite common receptor-ligand interactions, both receptor and ligand pairs share only limited sequence identity. A detailed molecular model of the extracellular Ig-like domain of human CD28 was constructed using a combination of different modeling methods. The model was based on the solution structure of CD152 and sequence comparison of the CD28/CD152 family. Assessment of the model revealed good stereochemical quality and sequence-structure compatibility. The CD28 model was used to map surface residues, N-linked glycosylation sites, and to compare residue conservation in CD28 and CD152. The location of N-linked glycosylation sites in CD28/CD152 restricts the surface area available for binding. Rigorous sequence conservation in CD28 and CD152 is limited to core IgSF consensus positions and surface residues implicated in ligand binding. Other surface residues vary greatly in CD28/CD152. Residues critical for ligand binding are surrounded by surface patches conserved only in either CD28 or CD152.


Bone mineral density distribution in the proximal femur and its relationship to morphologic factors in progressed unilateral hip osteoarthritis.
Although an adverse relationship between osteoporosis and osteoarthritis (OA) has been reported, it remains controversial. In most previous reports of OA, bone mineral density (BMD) changes in the subtrochanteric region have not been clarified, whilst BMD of the femoral neck and trochanteric region has been well investigated. In our current study, we investigated the BMD ratio compared to the contralateral side in the whole proximal femurs of hip OA patients. We aimed to clarify the morphologic factor that may influence these BMD ratios. We performed dual energy X-ray absorptiometry (DEXA) analysis of 69 hip joints from unilateral progressed OA cases. The minimum joint space, center edge angle, Sharp angle, acetabular head index, neck-shaft angle, and leg length discrepancy were also measured as radiographic factors. The correlation between BMD ratio and radiographic morphologic factors was then evaluated by logistic regression. The BMD ratio was higher in the femoral neck than in the distal region. In terms of radiographic factors, the neck-shaft angle was revealed to influence the decreased BMD ratio in the distal subtrochanteric part, whilst the leg length discrepancy and Sharp angle showed a relationship with the increased BMD ratio in the proximal neck region. The discrepancy in the BMD ratio between the femoral neck and the distal subtrochanteric region in the proximal femur is influenced by several morphologic factors.


A common pathway for genetic events leading to pheochromocytoma.
Mutations in VHL, RET, NF1, SDHB, SDHC, and SDHD can give rise to pheochromocytoma/paraganglioma. These different genetic lesions may all act by decreasing the activity of a 2-oxoglutarate-dependent oxygenase, SM-20/EglN3/PHD3, resulting in reduced apoptosis of neural crest cells during development.


Systemic administration of olygodeoxynucleotides with CpG motifs at priming phase reduces local Th2 response and late allergic rhinitis in BALB/c mice.
Oligodeoxynucleotides (ODN) with CpG motifs (CpG ODN) induce T helper (Th)1-type reaction. We aimed to evaluate the therapeutic effect of CpG ODN in the development of late allergic rhinitis induced by ovalbumin (OVA), which is one of Th2 diseaes, in BALB/c mice. Effects of a single dose of synthetic CpG-ODN (50 microg) intraperitoneally (i.p.) at the priming phase (on day 0) by OVA on the development of late eosinophilic rhinitis at respiratory areas were compared to the control mice treated with its vehicle (ODN without CpG motifs; 50 microg). Animals were again sensitized by OVA (on day 10) i.p., and 4 days after second sensitization animals were challenged by OVA intranasally (on day 14). Four days after challenge, eosinophilic reactions, nasal lesions and local cytokine values were examined. Compared to the control group, the CpG ODN-administration increased production of OVA-specific Th1 cytokine (interferon-gamma) and decreased productions of ovalubmin-specific Th2 cytokines [interleukin (IL)-5 and IL-13] in nasal cavity fluids, supernatants of splenocytes and/or sera. Also, eosinophilia and increased total IgE values were decreased in mice treated with the CpG ODN compared to the control group. Moreover, nasal lesions with infiltration of eosinophils were prominently reduced by the CpG ODN-treatment compared to the control mice. The present study suggests that the systemic administration of CpG ODN at the priming phase may reduce local OVA-specific Th2 responses, resulting in decreased nasal pathology in the late allergic eosinophilic rhinitis.


Bilayer Membrane Composed of Mineralized Collagen and Chitosan Cast Film Coated With Berberine-Loaded PCL/PVP Electrospun Nanofiber Promotes Bone Regeneration.
Bone defects are difficult to repair and reconstruct as bone regeneration remains technically challenging, with exogenous factors required to accelerate this process. Biodegradable synthetic scaffolds are promising materials for stimulating bone tissue repair. In this study, we investigated whether a bilayer membrane that includes mineralized collagen (MC) and chitosan (CS) delivering berberine (BER)-a typical Chinese herbal monomer-could promote bone healing in a rat model. An MC/CS cast film was coated with polycaprolactone (PCL)/polyvinylpyrrolidone (PVP) electrospun nanofibers loaded with BER, yielding the BER@PCL/PVP-MC/CS bilayer membrane. The 3-dimensional structure had nanofibers of uniform diameter and showed good hydrophilicity; the bilayer membrane showed favorable mechanical properties. BER@PCL/PVP-MC/CS enhanced the proliferation and attachment of MC3T3-E1 cells in vitro and induced bone regeneration when implanted into a rat femoral bone defect. These findings provide evidence that BER@PCL/PVP-MC/CS has clinical potential for effective bone repair.


ABA 9'-hydroxylation is catalyzed by CYP707A in Arabidopsis.
Abscisic acid (ABA) catabolism is important for regulating endogenous ABA levels. To date, most effort has focused on catabolism of ABA to phaseic acid (PA), which is generated spontaneously after 8'-hydroxylation of ABA by cytochrome P450s in the CYP707A subfamily. Neophaseic acid (neoPA) is another well-documented ABA catabolite that is produced via ABA 9'-hydroxylation, but the 9'-hydroxylase has not yet been defined. Here, we show that endogenous neoPA levels are reduced in loss-of-function mutants defective in CYP707A genes. In addition, in planta levels of both neoPA and PA are reduced after treatment of plants with uniconazole-P, a P450 inhibitor. These lines of evidence suggest that CYP707A genes also encode the 9'-hydroxylase required for neoPA synthesis. To test this, in vitro enzyme assays using microsomal fractions from CYP707A-expressing yeast strains were conducted and these showed that all four Arabidopsis CYP707As are 9'-hydroxylases, although this activity is minor. Collectively, our results demonstrate that ABA 9'-hydroxylation is catalyzed by CYP707As as a side reaction.


Manifestation of estrous behavior and subsequent progesterone concentration at timed-embryo transfer in cattle are positively associated with pregnancy success of recipients.
Plasma estradiol and progesterone (P4) concentrations during the peri-ovulatory period are positively correlated with pregnancy success in cattle. The aims of this study were to assess the effects of estrus occurrence and early diestrus P4 concentrations on pregnancy per timed-embryo transfer (P/TET). A total of 267 crossbred beef heifers [222 with corpus luteum (CL) and 45 without a CL but with a follicle >8mm at beginning of the estrous synchronization protocol) received an intra-vaginal P4 device and intramuscular administration of estradiol benzoate. Progesterone devices were removed 8 days later (Day 0), and heifers received d-cloprostenol, eCG and estradiol cypionate. Estrous behavior was monitored twice daily for 3 days after P4 device removal. Plasma P4 concentration was measured by radioimmunoassay at Day 7 and Day 9. At Day 9, heifers with a CL (n=236; i.e. submission rate of 85.5%; 236/276) undergoing TET received an in vitro-produced embryo. Heifers expressing a standing behavioral estrus had a greater P/TET than heifers that did not express a standing estrus [62.4% (106/170) compared with 47.0% (31/66)]. The probability of pregnancy was positively correlated with plasma P4 concentration at TET. When heifers were grouped by quartiles of P4 concentration at TET (Q1=0.64±0.16, Q2=1.70±0.04, Q3=2.90±0.07 and Q4=5.52±0.27ng/mL) the P/TET were 45.8% (Q1; 27/59)(c), 52.25% (Q2; 31/59)(bc), 66.1% (Q3; 39/59)(ab) and 67.8% (Q4; 40/59)(a). Additionally, heifers that became pregnant had greater P4 concentrations at TET (2.87±0.16ng/mL; n=137) than heifers that did not become pregnant (2.45±0.24ng/mL; n=99). No statistical difference was observed regarding P4 concentrations on Day 7, regardless of standing estrus or pregnancy status. In cattle, manifestation of estrous behavior and plasma P4 concentration at TET increase the probability of pregnancy in in vitro-produced embryo recipients.


Social interaction improves experimental stroke outcome.
Social interaction can have a profound effect on health. The purpose of the present study was to determine whether affiliative social interactions before and after stroke improve ischemic outcomes as assessed through histological analysis and behavioral assays. Male and female C57BL/6 mice were housed individually or with an ovariectomized female. Behavioral assessments were made 24 hours before 60 or 90 minutes of transient intraluminal middle cerebral artery occlusion (MCAO) or SHAM surgery and after 7 days of reperfusion. Two hours after behavioral testing on day 7, infarct size was determined by 2,3,5-triphenyltetrazolium histology, and blood samples were collected for assessment of corticosterone and C-reactive protein (CRP) concentrations. Pair housing significantly decreased infarct size and improved contralateral paw use in 60-minute MCAO males and 90-minute MCAO females compared with socially isolated cohorts. Housing condition had no significant effect on infarct size in females that underwent 60 minutes of MCAO, but pair housing was associated with improved contralateral paw use relative to socially isolated mice. In a separate cohort of males, intraischemic CRP concentration was significantly reduced in pair-housed males relative to isolated males. Affiliative interaction during the peri-ischemic period reduces intraischemic CRP concentration, decreases ischemic damage in male and female mice, and improves behavioral outcome.


The human lymphocyte micronucleus assay: a comparison between whole-blood and separated-lymphocyte cultures.
The induction of micronuclei (MN) by vincristine, mitomycin C and cyclophosphamide was compared in purified lymphocytes and in whole-blood cultures. With both assays, cytokinesis was blocked by cytochalasin B and MN were only scored in binucleate cells. The data suggest that whole-blood cultures may be considered a better experimental condition for the detection of MN induced by chemicals in vitro.


[Post-corona fatigue-a familiar picture in a new guise?]
Myalgic encephalitis or chronic fatigue syndrome (ME/CFS) has again come into focus as a result of coronavirus disease 2019 (COVID-19). Fundamentally problematic is the fact that ME/CFS is considered a separate entity; however, extreme fatigue is also a common symptom of an underlying disease. Our article aims to increase the acceptance of ME/CFS and extreme fatigue facing a symptomatology that is not fully understood, and to highlight the need for research, orientation for physicians, and counselling services for patients. Orientative research by focused information gathering. In various research projects, the hypothesis of post-infectious ME/CFS as an autoimmune disease could be confirmed. In general, the heterogeneity of diagnostic criteria as well as the variety of formulations to describe the symptomatology and different coding options make it difficult to clearly assign symptoms to a clinical picture. Exertion intolerance has been identified as a severe symptom of post-COVID-19 disorder. For this reason, recommendations in international guidelines are currently being revised, especially with regard to pacing. The implications for recommendations in tumor-related fatigue or due to multiple sclerosis are unclear. Against the background of a decreasing burden of disease due to increasing vaccination rates, research on fatigue should not only include viral diseases.


Generation of multi-scroll attractors without equilibria via piecewise linear systems.
In this paper, we present a new class of dynamical system without equilibria which possesses a multiscroll attractor. It is a piecewise-linear system which is simple, stable, displays chaotic behavior and serves as a model for analogous non-linear systems. We test for chaos using the 0-1 Test for Chaos from Gottwald and Melbourne [SIAM J. Appl. Dyn. Syst. 8(1), 129-145 (2009)].


Expression of ATP-sensitive potassium channels in human pregnant myometrium.
Potassium channels play critical roles in the regulation of cell membrane potential, which is central to the excitability of myometrium. The ATP-sensitive potassium (KATP) channel is one of the most abundant potassium channels in myometrium. The objectives of this study were to investigate the protein expression of KATP channel in human myometrium and determine the levels of KATP channel in lower and upper segmental myometrium before and after onset of labour. Both lower segmental (LS) and upper segmental (US) myometrial biopsies were collected at cesarean section from pregnant women not-in-labour (TNL) or in-labour (TL) at term. Protein expression level and cellular localization of four KATP channel subunits in US and LS myometrium were determined by Western blot analysis and immunohistochemistry, respectively. The contractile activity of myometrial strip was measured under isometric conditions. Four KATP channel subunits, namely Kir6.1, Kir6.2, SUR1 and SUR2B were identified in pregnant myometrium. While found in vascular myocytes, these subunits appear to be preferentially expressed in myometrial myocytes. Diazoxide, a KATP channel opener, inhibited the spontaneous contractility of pregnant myometrium, suggesting that the KATP channels are functional in human pregnant myometrium. Diazoxide was less potent in TL strips than that in TNL strips. Interestingly, expression of SUR1 was greater in TL than TNL tissues, although no differences were found for SUR2B in these two tissues. For both lower and upper segmental myometrium, Kir6.1 and Kir6.2 were less in TL compared with TNL tissues. Functional KATP channels are expressed in human pregnant myometrium. Down-regulation of Kir6.1 and Kir6.2 expression in myometrium may contribute to the enhanced uterine contractility associated with the onset of labour.


Semiautomated turbidimetric bioassay for the ionophore A23187.
The inhibitory effect of A23187 against Staphylococcus aureus was used to quantitate the activity of this ionophore in fermentation samples and in isolation and crystallization samples. The assay was shown to be rapid and reproducible.


Prevalence and Recovery from Newly Onset Anosmia and Ageusia in Covid 19 Patients at our Teritary Care Centre.
Anosmia (loss of smell) and Ageusia (loss of taste) are newly presenting independent symptoms in association with multiple symptoms such as Fever, Dry cough and Breathlessness in COVID 19 Patients.The Study of aims is to estimate Prevalence of Anosmia & Ageusia in Confirmed Covid 19 cases and to assess their Recovery rates. A Study conducted between June 2020 and September 2020 at a tertiary care COVID Dedicated hospital. Total 200 patients with age group 12-70 years Confirmed COVID 19 Positive Patients using RTPCR having mild to moderate symptoms were included. Patients were examined Clinically alongwith all safety measures to analyse prevalence & estimate their recoveries from sensory impairement using 10 item based DyNaCHRON questionnaire(concerned with taste and smell) at 10th, 14th & 21st Day. Out of 200 patients, Prevalence of Isolated Ageusia is 7%, Isolated Anosmia 4.5% and with Anosmia and Ageusia 4%. Complete Recovery regained in Ageusia within 14 days, while Rest all patients of Anosmia and both (Anosmia and Ageusia) within 21 days, Except 2 patients where long term Anosmia persists. Newly onset Anosmia and Ageusia are common in early stages of Covid 19 disease. They are Prevalent in mild to moderate symptomatic form of Diseases. Recovery in most cases is rapid and complete.Early Screening tests performed in suspected COVID 19 patients with loss of taste and smell sensation allows early diagnosis and treatment.


An immunochromatography assay for rapid antemortem diagnosis of dogs suspected to have canine distemper.
A new assay was developed for rapid and antemortem diagnosis of canine distemper (CD). This immunochromatography (IC)-based assay, which employs two monoclonal anti-CDV antibodies, was compared with nested PCR. When serial dilutions of purified CDV were tested, the CDV detection limits of the nested PCR and IC assays were 2 x 10(2)TCID(50)/ml and 5 x 10(2)TCID(50)/ml, respectively. Nasal irrigation fluid, conjunctival swabs, and blood lymphocytes from 66 dogs suspected to have CD were tested. Preliminary IC experiments revealed that the optimal sample volume and reaction time were 100 microl and 5 min, respectively. Relative to nested PCR, the sensitivity and specificity of the IC assay was maximal (100% and 100%, respectively) when conjunctival swabs were tested. This is significant because conjunctival swab specimens are easy to obtain in the early phase of CD infection. However, with blood lymphocytes and nasal samples, the IC assay was slightly less sensitive (89.7% and 85.7%, respectively) and specific (94.6% and 100%, respectively) than nested PCR. Since this novel IC assay does not require special instruments, it is a simple enough for dog owners to use. Since early detection of CD would allow appropriate treatment and quarantine to be instituted quickly, such a test would help reduce the morbidity and mortality associated with CD help to prevent its spread to other animals.


Clinical impact of intraoperative cone beam tomography and navigation for displaced acetabular fractures: a comparative study at medium-term follow-up.
The use of per-operative cone beam tomography imaging for displaced acetabular fractures yields increased post-operative articular reduction accuracy. This study evaluates the need for total hip replacement (THR) and hip-related functional outcomes in patients with displaced acetabular fractures treated with O-ARM guidance compared to those treated under C-ARM guidance. This is a prospective matched cohort study. Adult patients (35) with acetabular fractures operated under O-ARM guidance were included. These were matched (age, fracture type) to classically treated patients (35) from our data base. The primary outcome was the need for THR during three year follow-up period. Secondary outcomes were functional scores [Harris Hip score (HHS), Postel-Merle d'Aubigné (PMA)] and hip osteoarthritis grade at three year follow-up. Correlation between reduction gap and THR was evaluated. At three years, five patients were lost to follow-up in O-ARM group and four in control group. Two patients (6.66%) in the O-ARM group needed THR compared to eight patients in controls (25.80%) (p = 0.046). Hip X-ray osteoarthritis grade averaged 0.00 in patients without THR in O-ARM group compared to 0.22 in patients without THR in controls (p = 0.008). HHS averaged 95.79 in patients without THR in O-ARM group, compared to 93.82 in patients without THR in the control group (p = 0.41%). PMA averaged 17.25 in patients without THR in the O-ARM group compared to 17.04 in patients without THR in group 2 (p = 0.37). Evaluation of correlation between reduction gap and THR rate yielded OR = 1.22 (1.06-1.45). Increased accuracy in articular reduction, with per-operative three-dimensional control of impaction, in acetabular fractures led to significantly less need for THR in patients treated under O-ARM. Patients in both groups are comparable for functional outcomes because those with the lowest scores were offered THR. Per-operative cone beam guidance and navigation use are recommended in tertiary referral centres for acetabular trauma.


Assaying for Arginyltransferase Activity and Specificity by Peptide Arrays.
Here, we describe arginylation assays performed on peptide arrays immobilized on cellulose membranes via chemical synthesis. In this assay, it is possible to simultaneously compare arginylation activity on hundreds of peptide substrates to analyze the specificity of arginyltransferase ATE1 toward its target site(s) and the amino acid sequence context. This assay was successfully employed in prior studies to dissect the arginylation consensus site and enable predictions of arginylated proteins encoded in eukaryotic genomes.


Tumor necrosis factor-α enhances the transcription of Smad ubiquitination regulatory factor 1 in an activating protein-1- and Runx2-dependent manner.
Smad ubiquitination regulatory factor 1 (Smurf1) is an E3 ubiquitin ligase that negatively regulates osteoblast differentiation. Although tumor necrosis factor-α (TNF-α) has been shown to increase Smurf1 expression, the details of the regulatory mechanisms remain unclear. Here, we investigated the molecular mechanism by which TNF-α stimulates Smurf1 expression in C2C12 and primary cultured mouse calvarial cells. TNF-α treatment rapidly induced the activation of NF-κB and MAPKs. Smurf1 induction by TNF-α was blocked by the inhibition of JNK or ERK, while the inhibition of NF-κB and p38 MAPK had no effect on Smurf1 induction. TNF-α treatment or c-Jun overexpression enhanced the activity of a luciferase reporter that contained a 2.7 kb mouse Smurf1 promoter sequence. Site-directed mutagenesis of the Smurf1 reporter and chromatin immunoprecipitation analysis demonstrated that the activating protein-1 (AP-1) binding motif at -922 bp on the mouse Smurf1 promoter mediated TNF-α/JNK/AP-1-stimulated Smurf1 transcription. Interestingly, Smurf1 expression was not observed in Runx2-null mouse calvarial cells. When Runx2 was ectopically expressed in these cells, the basal and TNF-α-induced expression of Smurf1 was restored. Overexpression of Runx2 transactivated the Smurf1 promoter in a dose-dependent manner. Reporter and chromatin immunoprecipitation assays demonstrated that the Runx2-binding motif at -202 bp functioned in Runx2-mediated Smurf1 expression. ERK activation by TNF-α treatment or constitutively active MEK1 overexpression increased Smurf1 expression in a Runx2-dependent manner. These results suggest that the JNK/AP-1 and ERK/Runx2 signaling pathways mediate TNF-α-dependent Smurf1 transcription.


Methodological issues in estimating survival in patients with multiple primary cancers: an application to women with breast cancer as a first tumour.
Comparing survival of patients with a single tumour and patients with multiple primaries poses different methodological problems. In population based studies, where we cannot rely on detailed clinical information, the issue is disentangling the share of survival probability from the first and second cancer, and their compounded effect. We examined three hypotheses: A) the survival probability since the first tumour does not change with the occurrence of a second tumour; B) the probability of surviving a tumour does not change with the presence of a previous primary; C) the probabilities of surviving two subsequent primary tumours are independent (additivity hypothesis on mortality rates). We studied the survival probabilities modelling mortality rates according to hypotheses A), B) and C). Mortality rates were calculated using Aalen-Johansen estimators which allowed to discount for the lag-time survival before developing a second tumour. We applied this approach to a cohort of 436 women with breast cancer (BC) and a subsequent tumour in the resident population of Turin, Italy, between 1985 and 2002. We presented our results in term of a Standardised Mortality Ratio calculated (SMRAJ) after 10 years of follow-up. For hypothesis A we observed a significant excess mortality of 2.21 (95% C.I. 1.94 - 2.45). Concerning hypothesis B we found a not significant SMRAJ of 0.98 (95% C.I. 0.87 - 1.10). The additivity hypothesis (C) was not confirmed as it overestimated the risk of death, in fact SMRsAJ were all below 1: 0.75 (95% C.I. 0.66 - 0.84) for BC and all subsequent cancers, 0.72 (95% C.I. 0.55 - 0.94) for BC and colon-rectum cancer, 0.76 (95% C.I. 0.48 - 1.14) for BC and corpus uteri cancer (not significant). This method proved to be useful in disentangling the effect of different subsequent cancers on mortality. In our application it shows a worse long-term mortality for women with two cancers than that with BC only. However, the increase in mortality was lower than expected under the additivity assumption.


TDP43 proteinopathy is associated with aberrant DNA methylation in human amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by motor neurone (MN) degeneration and death. ALS can be sporadic (sALS) or familial, with a number of associated gene mutations, including C9orf72 (C9ALS). DNA methylation is an epigenetic mechanism whereby a methyl group is attached to a cytosine (5mC), resulting in gene expression repression. 5mC can be further oxidized to 5-hydroxymethylcytosine (5hmC). DNA methylation has been studied in other neurodegenerative diseases, but little work has been conducted in ALS. To assess differences in DNA methylation in individuals with ALS and the relationship between DNA methylation and TDP43 pathology. Post mortem tissue from controls, sALS cases and C9ALS cases were assessed by immunohistochemistry for 5mC and 5hmC in spinal cord, motor cortex and prefrontal cortex. LMNs were extracted from a subset of cases using laser capture microdissection. DNA from these underwent analysis using the MethylationEPIC array to determine which molecular processes were most affected. There were higher levels of 5mC and 5hmC in sALS and C9ALS in the residual lower motor neurones (LMNs) of the spinal cord. Importantly, in LMNs with TDP43 pathology there was less nuclear 5mC and 5hmC compared to the majority of residual LMNs that lacked TDP43 pathology. Enrichment analysis of the array data suggested RNA metabolism was particularly affected. DNA methylation is a contributory factor in ALS LMN pathology. This is not so for glia or neocortical neurones.


Redundant Fault Diagnosis for Photovoltaic Systems Based on an IRT Low-Cost Sensor.
In large solar farms, supervision is an exhaustive task, often carried out manually by field technicians. Over time, automated or semi-automated fault detection and prevention methods in large photovoltaic plants are becoming increasingly common. The same does not apply when talking about small or medium-sized installations, where the cost of supervision at such level would mean total economic infeasibility. Although there are prevention protocols by suppliers, periodic inspections of the facilities by technicians do not ensure that faults such as the appearance of hot-spots are detected in time. That is why, nowadays, the only way of continuous supervision of a small or medium installation is often carried out by unqualified people and in a purely visual way. In this work, the development of a low-cost system prototype is proposed for the supervision of a medium or small photovoltaic installation based on the acquisition and treatment of thermographic images, with the aim of investigating the feasibility of an actual implementation. The work focuses on the system's ability to detect hot-spots in supervised panels and successfully report detected faults. To achieve this goal, a low-cost thermal imaging camera is used for development, applying common image processing techniques, operating with OpenCV and MATLAB R2021b libraries. In this way, it is possible to demonstrate that it is achievable to successfully detect the hottest points of a photovoltaic (PV) installation with a much cheaper camera than the cameras used in today's thermographic inspections, opening up the possibilities of creating a fully developed low-cost thermographic surveillance system.


Segregation of Nogo66 receptors into lipid rafts in rat brain and inhibition of Nogo66 signaling by cholesterol depletion.
NogoA, a myelin-associated component, inhibits neurite outgrowth. Nogo66, a portion of NogoA, binds to Nogo66 receptor (NgR) and induces the inhibitory signaling. LINGO-1 and p75 neurotrophin receptor (p75), the low-affinity nerve growth factor receptor, are also required for NogoA signaling. However, signaling mechanisms downstream to Nogo receptor remain poorly understood. Here, we observed that NgR and p75 were colocalized in low-density membrane raft fractions derived from forebrains and cerebella as well as from cerebellar granule cells. NgR interacted with p75 in lipid rafts. In addition, disruption of lipid rafts by beta-methylcyclodextrin, a cholesterol-binding reagent, reduced the Nogo66 signaling. Our results suggest an important role of lipid rafts in facilitating the interaction between NgRs and provide insight into mechanisms underlying the inhibition of neurite outgrowth by NogoA.


Systemic Alterations of Cancer Cells and Their Boost by Polyploidization: Unicellular Attractor (UCA) Model.
Using meta-analyses, we introduce a unicellular attractor (UCA) model integrating essential features of the 'atavistic reversal', 'cancer attractor', 'somatic mutation', 'genome chaos', and 'tissue organization field' theories. The 'atavistic reversal' theory is taken as a keystone. We propose a possible mechanism of this reversal, its refinement called 'gradual atavism', and evidence for the 'serial atavism' model. We showed the gradual core-to-periphery evolutionary growth of the human interactome resulting in the higher protein interaction density and global interactome centrality in the UC center. In addition, we revealed that UC genes are more actively expressed even in normal cells. The modeling of random walk along protein interaction trajectories demonstrated that random alterations in cellular networks, caused by genetic and epigenetic changes, can result in a further gradual activation of the UC center. These changes can be induced and accelerated by cellular stress that additionally activates UC genes (especially during cell proliferation), because the genes involved in cellular stress response and cell cycle are mostly of UC origin. The functional enrichment analysis showed that cancer cells demonstrate the hyperactivation of energetics and the suppression of multicellular genes involved in communication with the extracellular environment (especially immune surveillance). Collectively, these events can unleash selfish cell behavior aimed at survival at all means. All these changes are boosted by polyploidization. The UCA model may facilitate an understanding of oncogenesis and promote the development of therapeutic strategies.


Topography-Guided Femto-LASIK in Virgin Eyes: Treating Manifest versus Measured Astigmatism.
To assess the stability, safety, predictability, and efficacy of topography-guided myopic Femto-LASIK with two different treatment protocols. Ebsar Eye center, Benha, Qalyopia, Egypt. Single-center, retrospective, COHORT control study. A total of 330 eyes enrolled in the study in group A and 322 eyes enrolled in group B underwent uncomplicated primary bilateral topography-guided Femto-LASIK. Group A was treated with the subjective clinical refraction; however, group B was treated with the modified refraction according to ALCON protocol. The mean preoperative refractive spherical equivalent (MRSE) was -4.85±1.90D and -5.0±1.93D in group A and B, respectively (P = 0.86), and a cylinder of -0.95±0.80 D and -0.92±0.81D, respectively. At the 12 months' postoperatively, the residual manifest SE within ± 0.5D was achieved by 82.86% of eyes in group A compared to 83.93% in group B. Of eyes, 92.06% had ≤0.5 astigmatism dioptre, while 100% of eyes had ≤1.0 astigmatism dioptre in group A (315 eyes); however, 91.80% of eyes had ≤0.5 astigmatism dioptre, while 100% of eyes had ≤1.0 astigmatism dioptre in group B. Topographic modification of the magnitude and axis of astigmatism treated using ALCON protocol when different from the clinical refraction may offer good refractive outcomes when we apply the Alcon precalculation considerations.


Intramolecular Friedel-Crafts alkylation with a silylium-ion-activated cyclopropyl group: formation of tricyclic ring systems from benzyl-substituted vinylcyclopropanes and hydrosilanes.
A trityl-cation-initiated annulation of benzyl-substituted vinylcyclopropanes (VCPs) with hydrosilanes is reported. Two Si-C(sp3) bonds and one C(sp2)-C(sp3) bond are formed in this process where an intramolecular 6-endo-tet Friedel-Crafts alkylation of a silylium-ion-activated cyclopropane ring is the rate-determining key step. The reaction mechanism is proposed based on computations and is in agreement with experimental observations. The new reaction leads to an unprecedented silicon-containing 6/6/5-fused ring system. A phenethyl-substituted VCP derivative yields another unknown tricycle having 6/6/6 ring fusion by reacting in a related but different way involving a 6-exo-tet ring closure.