On the 3D point clouds-palm and coconut trees data set extraction and their usages.
Drone image data set can be utilized for field surveying and image data collection which can be useful for analytics. With the current drone mapping software, useful 3D object reconstruction is possible. This research aims to learn the 3D data set construction process for trees with open-source software along with their usage. Thus, we research the tools used for 3D data set construction, especially in the agriculture field. Due to the growing open-source community, we demonstrate the case study of our palm and coconut data sets against the open-source ones. The methodology for achieving the point cloud data set was based on the tools: OpenDroneMap, CloudCompare, and Open3D. As a result, 40 palm trees and 40 coconut tree point clouds were extracted. Examples of the usages are provided in the area of volume estimation and graph analytics.


[Evaluation of the tranquilizing action of neurotropic drugs].
A comprehensive analysis of the cat brain electrograms (EG) by computer has revealed that the derivatives of 1,4-benzodiazepine (chlorodiazepoxide, diazepam) selectivity increase the modules of brain transients within the ranges of 8-12 and 10-12 Hz largely on the EG of the proreal gyrus and paleocortical structures and raise in the same spectral region the level of coherence between biopotentials of the limbic structures, displace the coefficients of asymmetry and excess unidimensional distributions of the instantaneous values of the EG amplitudes towards negative magnitudes. The indicators in question may be used as differential criteria of the tranquilizing effect of neurotropic substances. It is suggested that the increase in the modules of brain transients within the ranges of 8-10, 10-12 Hz during exposure to neurotropic substances is likely to mirror their central GABA-ergic mechanisms and to be an integral electrophysiological correlate of tranquilization as a psychophysiological state.


Characterization of the hyperpolarization-activated chloride current in dissociated rat sympathetic neurons.
1. Dissociated rat superior cervical ganglion (SCG) neurons have been shown to possess a hyperpolarization-activated inwardly rectifying chloride current. The current was not altered by changes in external potassium concentration, replacing external cations with NMDG (N-methyl-D-glucamine) or by addition of 10 mM caesium or barium ions. 2. The reversal potential of the current was altered by changing external anions. The anion selectivity of the current was Cl- > Br- > I- > cyclamate. All substituted permeant anions also blocked the current. 3. The current was blocked by DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid), 9AC (anthracene-9-carboxylic acid) and NPPB (5-nitro-2-(3-phenylpropylamino)benzoic acid) but was unaffected by SITS (4-acetamido-4'-isothiocyanatostilbene- 2,2'-disulphonic acid) and niflumic acid. The effective blockers were voltage dependent; DIDS and NPPB were more effective at depolarized potentials while 9AC was more effective at hyperpolarized potentials. 4. The current was enhanced by extracellular acidification and reduced by extracellular alkalinization. Reducing external osmolarity was without effect in conventional whole-cell recording but enhanced current amplitude in those perforated-patch recordings where little current was evident in control external solution. 5. The current in SCG neurons was blocked by external cadmium and zinc. ClC-2 chloride currents expressed in Xenopus oocytes were also sensitive to block by these divalent ions and by DIDS but the sensitivity of ClC-2 to block by cadmium ions was lower than that of the current in SCG neurons. 6. Reverse transcriptase-polymerase chain reaction (RT-PCR) experiments showed the presence of mRNA for ClC-2 in SCG neurons but not in rat cerebellar granule cells which do not possess a hyperpolarization-activated Cl- current. 7. The data suggest that ClC-2 may be functionally expressed in rat SCG neurons. This current may play a role in regulating the internal chloride concentration in these neurons and hence their response to activation of GABAA receptors.


Microenvironment Modulation of Single-Atom Ru in ZrSBA-15 for CO2 Hydrogenation to Formic Acid.
The chemical microenvironment modulation of active sites holds promise for facilitating their catalytic performance. Herein, single-atom Ru anchored by ZrSBA-15 modified with diverse organic amine groups has been fabricated and enabled CO2 hydrogenation to produce formic acid (FA) under mild conditions. However, the reaction cannot be achieved without the modification of organic amines. The functional groups as the microenvironment around Ru active sites effectively regulated the activity, in which Ru encapsulated in ZrSBA-15 bearing -NH2 groups exhibited the highest activity, with turnover number (TON) and turnover frequency (TOF) values of 505 and 64 h-1, respectively. Both characterization and experimental results validated that the functional group manipulated the adsorption capacity of the reactant, the electronic state of Ru and hydrophilicity/hydrophobicity of the materials, and thus the catalytic performance.


Trends in opioid-related crime incidents and comparison with opioid overdose outcomes in the United States.
The opioid epidemic in the United States remains a critically important public health issue and continues to worsen. While healthcare data and outcomes are commonly used to characterize the state of the epidemic and evaluate the impact of policy changes, criminal justice data is under-utilized in research despite its high relevance and unique role in the opioid crisis. Our objective is to understand temporal trends in opioid-related crime incidents and the comparability with the dynamic patterns in health-related outcomes. We used incident-level crime data from the National Incident-Based Reporting System (NIBRS) during 2005-2018. We identified all incidents involving opioids, which were grouped by opioid type (illicit and prescription opioids), and by drug-related criminal activity (possession and distribution). We estimated annual opioid-related crime incident rates per 100,000 residents. Joinpoint analysis was performed to examine the significant changes in the temporal trends of crime incident rates. We examined the association between opioid-related crime incidents and health outcomes using state fixed effects regression models. Among the NIBRS covered population, incident rates of all opioid-related crimes increased significantly from 32.0 to 91.4 per 100,000 between 2005 and 2016, followed by a moderate decrease to 78.3 per 100,000 by 2018. The initial increase in incident rates was predominantly driven by prescription opioid-related incidents which increased by 19.6% per year from 2005 to 2010. Between 2010 and 2015, most of the increase came from illicit opioid-related incidents which accelerated to an increase of 21.6% per year. Opioid-related crime incident rates were found to be significantly and positively associated with rates of opioid-related emergency department visits, inpatient hospitalization, and overdose mortality. Crime data describe temporal trends and shifting patterns in the opioid epidemic that are highly consistent with health-related data. Criminal justice data could be a potentially powerful tool to understand the changing landscape of opioid and substance use.


Molecular diagnosis of tuberculosis: current clinical validity and future perspectives.
The rapid development and availability of a variety of new molecular genetic technologies present the clinician with an array of options for the accurate diagnosis of infectious diseases. This is particularly the case for tuberculosis, since molecular methods have been rapidly introduced into all working areas of the mycobacteriology laboratory. Nucleic acid amplification methods to detect Mycobacterium tuberculosis in clinical specimens are increasingly used as a tool to diagnose tuberculosis. The bulk of recently available data from clinical evaluations performed under routine laboratory conditions indicate that these molecular methods are rapid and sensitive, but yet inferior, to culture with regard to sensitivity and specificity. Therefore, until gene amplification tests have proved to be reliable and quality control procedures exist, their clinical validity remains controversial. Consequently, definition of selected clinical applications of gene amplification to routine diagnosis of tuberculosis is important and need to be discussed. This review will focus on the clinical role of molecular methods in the direct detection and diagnosis of M. tuberculosis in clinical samples. In addition, molecular genetic approaches designed to determine drug susceptibility and to discriminate strains below the species level will be outlined and discussed in terms of their current and future clinical applicability.


Use of inhaled corticosteroids and risk of acquiring Pseudomonas aeruginosa in patients with chronic obstructive pulmonary disease.
Inhaled corticosteroids (ICS) are commonly used to treat COPD and are associated with increased risk of pneumonia. The aim of this study was to assess if accumulated use of ICS is associated with a dose-dependent risk of a positive airway culture with Pseudomonas aeruginosa in patients with COPD. We conducted a multiregional epidemiological cohort study including Danish COPD patients followed in outpatient clinics during 2010-2017. ICS use was categorised based on accumulated prescriptions redeemed 365 days prior to cohort entry. Cox proportional hazard regression model was used to estimate the risk of acquiring P. aeruginosa. Propensity score matched models were used as sensitivity analyses. A total of 21 408 patients were included in the study, of which 763 (3.6%) acquired P. aeruginosa during follow-up. ICS use was associated with a dose-dependent risk of P. aeruginosa (low ICS dose: HR 1.38, 95% CI 1.03 to 1.84, p=0.03; moderate ICS dose: HR 2.16, 95% CI 1.63 to 2.85, p<0.0001; high ICS dose: HR 3.58, 95% CI 2.75 to 4.65, p<0.0001; reference: no ICS use). A propensity matched model confirmed the results (high ICS dose compared with no/low/moderate ICS dose: HR 2.05, 95% CI 1.76 to 2.39, p p<0.0001). Use of ICS in patients with COPD followed in Danish outpatient clinics was associated with a substantially increased and dose-dependent risk of acquiring P. aeruginosa. Caution should be taken when administering high doses of ICS in severely ill patients with COPD. These results should be confirmed in comparable cohorts and other settings.


Inositol polyphosphates and target of rapamycin kinase signalling govern photosystem II protein phosphorylation and photosynthetic function under light stress in Chlamydomonas.
Stress and nutrient availability influence cell proliferation through complex intracellular signalling networks. In a previous study it was found that pyro-inositol polyphosphates (InsP7 and InsP8 ) produced by VIP1 kinase, and target of rapamycin (TOR) kinase signalling interacted synergistically to control cell growth and lipid metabolism in the green alga Chlamydomonas reinhardtii. However, the relationship between InsPs and TOR was not completely elucidated. We used an in vivo assay for TOR activity together with global proteomic and phosphoproteomic analyses to assess differences between wild-type and vip1-1 in the presence and absence of rapamycin. We found that TOR signalling is more severely affected by the inhibitor rapamycin in a vip1-1 mutant compared with wild-type, indicating that InsP7 and InsP8 produced by VIP1 act independently but also coordinately with TOR. Additionally, among hundreds of differentially phosphorylated peptides detected, an enrichment for photosynthesis-related proteins was observed, particularly photosystem II proteins. The significance of these results was underscored by the finding that vip1-1 strains show multiple defects in photosynthetic physiology that were exacerbated under high light conditions. These results suggest a novel role for inositol pyrophosphates and TOR signalling in coordinating photosystem phosphorylation patterns in Chlamydomonas cells in response to light stress and possibly other stresses.


Older adults and hearing help-seeking behaviors.
To review the current literature on help seeking for hearing health care among older adults. The authors conducted a literature review regarding help seeking for hearing-related communication difficulties as well as for other chronic medical conditions. Untreated hearing loss can lead to numerous negative secondary consequences; uptake and use of hearing aids remain low, despite the fact that hearing aids provide an effective treatment option for older adults with hearing loss. The authors describe models relevant to understanding the help-seeking and decision-making behaviors of older adults with hearing loss and discuss recommendations for future research. Because of the considerable overlap in factors associated with help-seeking behaviors across chronic medical conditions and because help-seeking behaviors are complex, help seeking should be examined within the framework of a multifactorial model, such as the health belief model or the transtheoretical stages of change model.


Multi-omics analyses demonstrate the modulating role of gut microbiota on the associations of unbalanced dietary intake with gastrointestinal symptoms in children with autism spectrum disorder.
Our previous work revealed that unbalanced dietary intake was an important independent factor associated with constipation and gastrointestinal (GI) symptoms in children with autism spectrum disorder (ASD). Growing evidence has shown the alterations in the gut microbiota and gut microbiota-derived metabolites in ASD. However, how the altered microbiota might affect the associations between unbalanced diets and GI symptoms in ASD remains unknown. We analyzed microbiome and metabolomics data in 90 ASD and 90 typically developing (TD) children based on 16S rRNA and untargeted metabolomics, together with dietary intake and GI symptoms assessment. We found that there existed 11 altered gut microbiota (FDR-corrected P-value <0.05) and 397 altered metabolites (P-value <0.05) in children with ASD compared with TD children. Among the 11 altered microbiota, the Turicibacter, Coprococcus 1, and Lachnospiraceae FCS020 group were positively correlated with constipation (FDR-corrected P-value <0.25). The Eggerthellaceae was positively correlated with total GI symptoms (FDR-corrected P-value <0.25). More importantly, three increased microbiota including Turicibacter, Coprococcus 1, and Eggerthellaceae positively modulated the associations of unbalanced dietary intake with constipation and total GI symptoms, and the decreased Clostridium sp. BR31 negatively modulated their associations in ASD children (P-value <0.05). Together, the altered microbiota strengthens the relationship between unbalanced dietary intake and GI symptoms. Among the altered metabolites, ten metabolites derived from microbiota (Turicibacter, Coprococcus 1, Eggerthellaceae, and Clostridium sp. BR31) were screened out, enriched in eight metabolic pathways, and were identified to correlate with constipation and total GI symptoms in ASD children (FDR-corrected P-value <0.25). These metabolomics findings further support the modulating role of gut microbiota on the associations of unbalanced dietary intake with GI symptoms. Collectively, our research provides insights into the relationship between diet, the gut microbiota, and GI symptoms in children with ASD.


miR-93 suppresses proliferation and colony formation of human colon cancer stem cells.
To identify differentially expressed microRNAs (miRNAs) in human colon cancer stem cells (SW1116csc) and study their function in SW1116csc proliferation. SW1116csc were isolated from the human colon cancer cell line, SW1116 and cultured in serum-free medium. A miRNA microarray was used to detect differential expression profiles of miRNAs in SW1116csc and SW1116 cells. Real-time quantitative polymerase chain reaction (PCR) was performed to verify the differential expression of candidate miRNAs obtained from the microarray. Target mRNAs of differentially expressed miRNAs were predicted with target prediction tools. miRNA expression plasmids were transfected into SW1116csc using Lipofectamine 2000 reagent. Cell proliferation curves were generated with trypan blue staining, and the colony formation rate of transfected cells was measured with the soft agar colony formation assay. Expression of target mRNAs and proteins from differentially expressed miRNAs were detected using reverse transcription (RT)-PCR and western blotting. Compared with expression in SW1116 cells, 35 miRNAs (including hsa-miR-192, hsa-miR-29b, hsa-miR-215, hsa-miR-194, hsa-miR-33a and hsa-miR-32) were upregulated more than 1.5-fold, and 11 miRNAs (including hsa-miR-93, hsa-miR-1231, hsa-miRPlus-F1080, hsa-miR-524-3p, hsa-miR-886-3p and hsa-miR-561) were downregulated in SW1116csc. The miRNA microarray results were further validated with quantitative RT-PCR. miR-93 was downregulated, and its predicted mRNA targets included BAMBI, CCND2, CDKN1A, HDAC8, KIF23, MAP3K9, MAP3K11, MYCN, PPARD, TLE4 and ZDHHC1. Overexpressed miR-93 significantly inhibited cell proliferation and colony formation by SW1116csc. Furthermore, miR-93 negatively regulated the mRNA and protein levels of HDAC8 and TLE4. Some miRNAs were differentially expressed during differentiation of SW1116csc into SW1116 cells. miR-93 may inhibit SW1116csc proliferation and colony formation.


Cytolysins augment superantigen penetration of stratified mucosa.
Staphylococcus aureus and Streptococcus pyogenes colonize mucosal surfaces of the human body to cause disease. A group of virulence factors known as superantigens are produced by both of these organisms that allows them to cause serious diseases from the vaginal (staphylococci) or oral mucosa (streptococci) of the body. Superantigens interact with T cells and APCs to cause massive cytokine release to mediate the symptoms collectively known as toxic shock syndrome. In this study we demonstrate that another group of virulence factors, cytolysins, aid in the penetration of superantigens across vaginal mucosa as a representative nonkeratinized stratified squamous epithelial surface. The staphylococcal cytolysin alpha-toxin and the streptococcal cytolysin streptolysin O enhanced penetration of toxic shock syndrome toxin-1 and streptococcal pyrogenic exotoxin A, respectively, across porcine vaginal mucosa in an ex vivo model of superantigen penetration. Upon histological examination, both cytolysins caused damage to the uppermost layers of the vaginal tissue. In vitro evidence using immortalized human vaginal epithelial cells demonstrated that although both superantigens were proinflammatory, only the staphylococcal cytolysin alpha-toxin induced a strong immune response from the cells. Streptolysin O damaged and killed the cells quickly, allowing only a small release of IL-1beta. Two separate models of superantigen penetration are proposed: staphylococcal alpha-toxin induces a strong proinflammatory response from epithelial cells to disrupt the mucosa enough to allow for enhanced penetration of toxic shock syndrome toxin-1, whereas streptolysin O directly damages the mucosa to allow for penetration of streptococcal pyrogenic exotoxin A and possibly viable streptococci.


Combination therapy for carbapenem-resistant Gram-negative bacteria.
Carbapenem-resistant Gram-negative bacteria (CR-GNB) represent an increasing hazard in healthcare settings. A central question concerning the treatment of invasive infections caused by CR-GNB involves the use of combination therapy. Potential advantages of combination therapy include improved efficacy due to synergy, while the disadvantages include adverse events and increased antibiotic use with a potential drive towards resistance. Several observational studies have examined whether combination therapy offers an advantage over colistin/polymyxin monotherapy. We highlight the inherent limitations of these studies related to their observational design and sample size to show why they do not at present provide an answer to the question of combination versus monotherapy. This distinction is important to guide clinical practice until solid evidence has been obtained and to enable the recruitment of patients into randomized controlled trials. A few randomized controlled trials examining specific combinations have recently been completed or are ongoing. Currently, however, there is no evidence-based support for most combination therapies against CR-GNB, including colistin/carbapenem combination therapy.


[Relationship between the insulin-stimulated disappearance of inorganic phosphate in vitro and the effect of insulin on creatine accumulation in rat skeletal muscle plasma membranes].
Total content of creatine was increased in unfractionated membrane fractions within 5-30 min after incubation at 30 degrees of the repeatedly washed and suspended in 0.25 M sucrose plasmatic membranes, isolated from rat skeletal muscle, using a medium containing tris-HCl buffer, rH 7.5, AMP and ADP, Mg2+, creatine, inorganic phosphate, NaF, NAD or NAD-dependent system of lactate oxidation (NAD, lactate, cytochrome c and O2) in presence of insulin (4 mg/ml). This increase in total content of creatine, stimulated by insulin, was accompanied by rapid diappearance of inorganic phosphate, which was especially distinct within 1 min of inculation at 30 degrees. Nonactive derivative of insulin (desoctapeptide insulin) did not affect the accumulation of creatine. The effect of inculin on accumulation of creatine was abolished by means of addition of EGTA into the medium or by excluding of Mg2+-AMP and of inorganic phosphate from the medium. The phenomenon observed might be related to synthesis of ATP, required for the physiological activity of insulin during transport of creatine across the biomembranes.


EUS-guided versus percutaneous liver biopsy: A prospective randomized clinical trial.
Prospective studies comparing EUS-guided liver biopsy (EUS-LB) to percutaneous LB (PC-LB) are scarce. We compared the efficacy and safety of EUS-LB with those of PC-LB in a prospective randomized clinical trial. Between 2020 and 2021, patients were enrolled and randomized (1:1 ratio). The primary outcome was defined as the proportion of patients with ≥11 complete portal tracts (CPTs). The sample size (n = 80) was calculated based on the assumption that 60% of those in the EUS-LB and 90% of those in the PC-LB group will have LB with ≥11 CPTs. The secondary outcomes included proportion of patients in whom a diagnosis was established, number of CPTs, pain severity (Numeric Rating Scale-Pain Intensity), duration of hospital stay, and adverse events. Eighty patients were enrolled (median age, 53 years); 67.5% were female. Sixty percent of those in the EUS-LB and 75.0% of those in the PC-LB group met the primary outcome (P = 0.232). The median number of CPTs was higher in the PC-LB (17 vs 13; P = 0.031). The proportion of patients in whom a diagnosis was established was similar between the groups (92.5% [EUS-LB] vs 95.0% [PC-LB]; P = 1.0). Patients in the EUS-LB group had less pain severity (median Numeric Rating Scale-Pain Intensity, 2.0 vs 3.0; P = 0.003) and shorter hospital stay (2.0 vs 4.0 hours; P < 0.0001) compared with the PC-LB group. No patient experienced a serious adverse event. EUS-guided liver biopsy was safe, effective, better tolerated, and associated with a shorter hospital stay.


Diverse roles for the Notch receptor in the development of D. melanogaster.
Notch proteins appear to be involved in cell fate commitments with deep evolutionary roots. Homologues have been shown to play key roles in the development of nematodes, insects, amphibia, and mammals. Activity of the Notch receptor has been observed in the patterning of ectoderm, mesoderm, and endoderm, indicating an origin prior to the functional differentiation of these germ layers. To understand how a single receptor can participate so widely in development, we have been examining the role of specific extracellular segments of Notch. Early studies of mutations affecting widely separated EGF-like elements of Notch first raised the possibility for interaction with multiple ligands. Biochemical approaches, and exhaustive structure function studies in transgenic Drosophila are beginning to reveal how this receptor is activated, and point to a range of physical interactions with other proteins.


Development of a Zulu speech reception threshold test for Zulu first language speakers in Kwa Zulu-Natal.
The measurement of speech reception threshold (SRT) is best evaluated in an individual's first language. The present study focused on the development of a Zulu SRT word list, according to adapted criteria for SRT in Zulu. The aim of this paper is to present the process involved in the development of the Zulu word list. In acquiring the data to realize this aim, 131 common bisyllabic Zulu words were identified by two Zulu speaking language interpreters and two tertiary level educators. Eighty two percent of these words were described as bisyllabic verbs. Thereafter using a three point Likert scale, 58 bisyllabic verbs were rated by 5 linguistic experts as being familiar, phonetically dissimilar and being low tone verbs. According to the Kendall's co-efficient of concordance at 95% level of confidence the agreement among the raters was good for each criterion. The results highlighted the importance of adapting the criteria for SRT to suit the structure of the language. An important research implication emerging from the study is the theoretical guidelines proposed for the development of SRT material in other African Languages. Furthermore, the importance of using speech material appropriate to the language has also being highlighted. The developed SRT word list in Zulu is applicable to the adult Zulu First Language Speaker in KZN.


Drowning in disinfection byproducts? Assessing swimming pool water.
Disinfection is mandatory for swimming pools: public pools are usually disinfected by gaseous chlorine or sodium hypochlorite and cartridge filters; home pools typically use stabilized chlorine. These methods produce a variety of disinfection byproducts (DBPs), such as trihalomethanes (THMs), which are regulated carcinogenic DBPs in drinking water that have been detected in the blood and breath of swimmers and of nonswimmers at indoor pools. Also produced are halogenated acetic acids (HAAs) and haloketones, which irritate the eyes, skin, and mucous membranes; trichloramine, which is linked with swimming-pool-associated asthma; and halogenated derivatives of UV sun screens, some of which show endocrine effects. Precursors of DBPs include human body substances, chemicals used in cosmetics and sun screens, and natural organic matter. Analytical research has focused also on the identification of an additional portion of unknown DBPs using gas chromatography (GC)/mass spectrometry (MS) and liquid chromatography (LC)/MS/MS with derivatization. Children swimmers have an increased risk of developing asthma and infections of the respiratory tract and ear. A 1.6-2.0-fold increased risk for bladder cancer has been associated with swimming or showering/bathing with chlorinated water. Bladder cancer risk from THM exposure (all routes combined) was greatest among those with the GSTT1-1 gene. This suggests a mechanism involving distribution of THMs to the bladder by dermal/inhalation exposure and activation there by GSTT1-1 to mutagens. DBPs may be reduced by engineering and behavioral means, such as applying new oxidation and filtration methods, reducing bromide and iodide in the source water, increasing air circulation in indoor pools, and assuring the cleanliness of swimmers. The positive health effects gained by swimming can be increased by reducing the potential adverse health risks.


A docking model based on mass spectrometric and biochemical data describes phage packaging motor incorporation.
The molecular mechanism of scaffolding protein-mediated incorporation of one and only one DNA packaging motor/connector dodecamer at a unique vertex during lambdoid phage assembly has remained elusive because of the lack of structural information on how the connector and scaffolding proteins interact. We assembled and characterized a phi29 connector-scaffolding complex, which can be incorporated into procapsids during in vitro assembly. Native mass spectrometry revealed that the connector binds at most 12 scaffolding molecules, likely organized as six dimers. A data-driven docking model, using input from chemical cross-linking and mutagenesis data, suggested an interaction between the scaffolding protein and the exterior of the wide domain of the connector dodecamer. The connector binding region of the scaffolding protein lies upstream of the capsid binding region located at the C terminus. This arrangement allows the C terminus of scaffolding protein within the complex to both recruit capsid subunits and mediate the incorporation of the single connector vertex.


Immobilized bacteria with pH-response hydrogel for self-healing of concrete.
Concrete is significant for construction. A problem in application is the appearance of cracks that will damage its strength. An autogenous crack-healing mechanism based on bacteria receives increasing attention in recent years. The bacteria are able to form calcium carbonate (CaCO3) precipitations in suitable conditions to protect and reinforce the concrete. However, a large number of spores are crushed in aged specimens, resulting in a loss of viability. A new kind of hydrogel crosslinked by alginate, chitosan and calcium ions was introduced in this study. It was observed that the addition of chitosan improved the swelling properties of calcium alginate. Opposite pH response to calcium alginate was observed when the chitosan content in the solution reached 1.0%. With an addition of 1.0% chitosan in hydrogel beads, 10.28% increase of compressive strength and 13.79% increase of flexural strength to the control were observed. The results reveal self-healing properties of concretes. A healing crack of 4 cm length and 1 mm width was observed when using cement PO325, with the addition of bacterial spores (2.54-3.07 × 105/cm3 concrete) encapsulated by hydrogel containing no chitosan.


Direct comparison of electron transfer properties of two distinct semisynthetic triads with non-protein based triad: unambiguous experimental evidences on protein matrix effects.
In order to understand the roles of protein matrix in electron transfer processes (ET) within biological systems, a heme-based donor (Zn-heme: ZnPP)-sensitizer (Ru2+(bpy)3)-acceptor (cyclic viologen: BXV4+) triad 1 was used as a probe molecule. Two semi-synthetic systems, Cyt-b562(1) and Mb(1), in which the triad is incorporated into cytochrome b562 (Cyt-b562) or into myoglobin (Mb), were constructed by cofactor reconstitution. These two semi-synthetic proteins were compared with the triad itself (i.e., without the protein matrix) using absorption spectroscopy, steady state emission and excitation studies, laser flash photolysis experiments, and molecular modeling. Photoexcitation of the ZnPP moiety of Cyt-b562(1) or Mb(1) leads to a direct ET from the triplet state of ZnPP state (3ZnPP) to BXV4+ to generate a final charge-separated (CS) state, Cyt-b562(Zn+)-Ru2+-BXV3+* or Mb(Zn+)-Ru2+-BXV3+*. On the other hand, direct ET from the excited ZnPP moiety to the BXV4+ moiety is also involved in 1 in the absence of the protein matrix, but the excited state of ZnPP involved is not 3ZnPP, but the singlet excited state (1ZnPP) in this pathway. When the Ru2+(bpy)3 moiety of Cyt-b562(1) or Mb(1) is excited, a stepwise ET relay occurs with the ion-pair, Cyt-b562(Zn)-Ru3+-BXV3+* or Mb(Zn)-Ru3*-BXV3+*, as an intermediate, leading to the same final CS state as that generated in the direct ET pathway. The lifetimes of the corresponding final CS states were determined to be 300 ns for 1 in the absence of the protein matrix, 600-900 ns for Cyt-b562(1) and 1.1-18 micros for Mb(1), the values of which are greatly affected by the protein matrix. Molecular modeling study of the three systems consistently explained the differences of their photophysical behavior.


Impaired insight in Alzheimer disease: association with cognitive deficits, psychiatric symptoms, and behavioral disturbances.
The purpose of this study was to evaluate symptoms associated with impaired insight in patients with Alzheimer disease (AD). Although unawareness of deficits is common in AD, the relation of awareness to psychiatric and behavioral disturbances has not been extensively studied. We conducted a cross-sectional investigation of 91 patients with probable AD according to the criteria of the National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association. Awareness of cognitive and functional deficits was measured with the Inaccurate Insight item from the Neurobehavioral Rating Scale. Psychiatric and behavioral symptoms were measured using factor scores and individual items from the Neurobehavioral Rating Scale. Global cognitive deficits were measured using the Mini-Mental State Examination (MMSE). Stepwise regression analysis showed that insight was associated with MMSE score, depression/anxiety factor score, and agitation/disinhibition factor score. Variables not associated with awareness of deficits included patient age, behavioral retardation factor score, verbal output disturbance factor score, and psychosis factor score. Post hoc analyses showed a positive relation (i.e., greater insight, more symptomatology) between deficit awareness and symptoms of depressed mood and anxiety. There was a negative relation (i.e., greater insight, less symptomatology) between insight and symptoms of hostility, agitation, inattention, and tension. In a follow-up stepwise regression analysis, increased deficit awareness was associated with a higher MMSE score, greater depressed mood, and decreased agitation. These findings suggest that patients with AD may experience symptoms of depressed mood in relation to increased awareness of decrements in functioning. The data also indicate that patients with poor insight demonstrate greater agitated behavior. Consistent with previous research, impaired insight was higher in the later stages of the illness.


Use of clinical and impairment-based tests to predict falls by community-dwelling older adults.
Few tests have been found to be strongly predictive of falls in community-dwelling older adults. The purpose of this study was to determine whether data from 5 balance tests-combined with data regarding fall history, number of medications, dizziness, visual problems, use of an assistive device, physical activity level, sex, and age-could predict falls in community-dwelling older adults who were independent. Ninety-nine community-dwelling older adults aged 65 to 90 years (X =74.02, SD=5.64) were tested. Subjects were tracked for falls over a 1-year period following testing. Impairment-based tests, which are tests that attempt to specifically identify which sensory systems are impaired or how motor control is impaired (eg, speed, accuracy of movement), were the Modified Clinical Tests of Sensory Interaction for Balance (Modified CTSIB) and the 100% Limits of Stability Test, both of which were done on the Balance Master 6.1. Performance-based tests, which are functional tests that identify functional limitations without necessarily identifying their causes, were the Berg Balance Scale, the Timed "Up Go" Test, and the Dynamic Gait Index. Demographic and health data included age, sex, number of medications, physical activity level, presence of dizziness, vision problems, and history of falls over the previous year. Logistic regression was used to determine which combinations of data from balance tests, demographics, and health factors were predictive of falls. Two models-(1) the "standing on a firm surface with eyes closed" (FEC) condition of the Modified CTSIB and (2) the FEC combined with age and sex-were predictive of falls, but predicted only 1 and 2 subjects who were at risk for falling, respectively, out of 20 people who were at risk for falling. Five balance tests combined with health and demographic factors did not predict falls in a sample of community-dwelling older adults who were active and independent.


Non-technical skills of surgeons and anaesthetists in simulated operating theatre crises.
Deficiencies in non-technical skills (NTS) have been increasingly implicated in avoidable operating theatre errors. Accordingly, this study sought to characterize the impact of surgeon and anaesthetist non-technical skills on time to crisis resolution in a simulated operating theatre. Non-technical skills were assessed during 26 simulated crises (haemorrhage and airway emergency) performed by surgical teams. Teams consisted of surgeons, anaesthetists and nurses. Behaviour was assessed by four trained raters using the Non-Technical Skills for Surgeons (NOTSS) and Anaesthetists' Non-Technical Skills (ANTS) rating scales before and during the crisis phase of each scenario. The primary endpoint was time to crisis resolution; secondary endpoints included NTS scores before and during the crisis. A cross-classified linear mixed-effects model was used for the final analysis. Thirteen different surgical teams were assessed. Higher NTS ratings resulted in significantly faster crisis resolution. For anaesthetists, every 1-point increase in ANTS score was associated with a decrease of 53·50 (95 per cent c.i. 31·13 to 75·87) s in time to crisis resolution (P < 0·001). Similarly, for surgeons, every 1-point increase in NOTSS score was associated with a decrease of 64·81 (26·01 to 103·60) s in time to crisis resolution in the haemorrhage scenario (P = 0·001); however, this did not apply to the difficult airway scenario. Non-technical skills scores were lower during the crisis phase of the scenarios than those measured before the crisis for both surgeons and anaesthetists. A higher level of NTS of surgeons and anaesthetists led to quicker crisis resolution in a simulated operating theatre environment.


High yield expression of lipase A from Candida antarctica in the methylotrophic yeast Pichia pastoris and its purification and characterisation.
The current investigation focuses on shedding further light on the characteristics of lipase A from Candida antarctica (CalA), which has attracted growing attention in its suitability for industrial applications. CalA was functionally expressed in the methylotrophic yeast Pichia pastoris, purified and characterised. A classical fed-batch process and a semi-continuous process were developed and tested with regard to their yield capacity. Lipase concentrations of 0.88 and 0.55 g l(-1) were obtained using the fed-batch and semi-continuous processes, respectively. The semi-continuous process reaches a total activity of 10,233,000 U and so surpasses the fed-batch process reaching 7,530,000 U. The purified enzyme showed highest activity between 50 and 70 degrees C at pH 7.0 and a preference for short-chain triglycerides (C4-C8). Significantly reduced activity was observed in the presence of hydrophilic esters.


Expression and characterization of Drosophila signal peptide peptidase-like (sppL), a gene that encodes an intramembrane protease.
Intramembrane proteases of the Signal Peptide Peptidase (SPP) family play important roles in developmental, metabolic and signaling pathways. Although vertebrates have one SPP and four SPP-like (SPPL) genes, we found that insect genomes encode one Spp and one SppL. Characterization of the Drosophila sppL gene revealed that the predicted SppL protein is a highly conserved structural homolog of the vertebrate SPPL3 proteases, with a predicted nine-transmembrane topology, an active site containing aspartyl residues within a transmembrane region, and a carboxy-terminal PAL domain. SppL protein localized to both the Golgi and ER. Whereas spp is an essential gene that is required during early larval stages and whereas spp loss-of-function reduced the unfolded protein response (UPR), sppL loss of function had no apparent phenotype. This was unexpected given that genetic knockdown phenotypes in other organisms suggested significant roles for Spp-related proteases.


The blood-CSF-brain route of neurological disease: The indirect pathway into the brain.
The brain is protected by the endothelial blood-brain barrier (BBB) that limits the access of micro-organisms, tumour cells, immune cells and autoantibodies to the parenchyma. However, the classic model of disease spread across a disrupted BBB does not explain the focal distribution of lesions seen in a variety of neurological diseases and why lesions are frequently adjacent to the cerebrospinal fluid (CSF) spaces. We have critically reviewed the possible role of a blood-CSF-brain route as a disease entry pathway into the brain parenchyma. The initial step of this pathway is the transfer of pathogens or immune components from the blood into the CSF at the choroid plexuses, where the blood-CSF barrier (BCSFB) is located. The flow of CSF results in disease dissemination throughout the CSF spaces. Access to the brain parenchyma from the CSF can then occur across the ependymal layer at the ventricular surface or across the pial-glial barrier of the subarachnoid space and the Virchow-Robin spaces. We have reviewed the anatomy and physiology of the blood-CSF-brain pathway and the brain barriers controlling this process. We then summarised the evidence supporting this brain entry route in a cross-section of neurological diseases including neuromyelitis optica, multiple sclerosis, neurosarcoidosis, neuropsychiatric lupus, cryptococcal infection and both solid and haematological tumours. This summary highlights the conditions that share the blood-CSF-brain pathway as a pathogenetic mechanism. These include the characteristic proximity of lesions to CSF, evidence of disruption of the brain barriers and the identification of significant pathology within the CSF. An improved understanding of pathological transfer through the CSF and across all brain barriers will inform on more effective and targeted treatments of primary and secondary diseases of the central nervous system.


Simultaneous tensor decomposition and completion using factor priors.
The success of research on matrix completion is evident in a variety of real-world applications. Tensor completion, which is a high-order extension of matrix completion, has also generated a great deal of research interest in recent years. Given a tensor with incomplete entries, existing methods use either factorization or completion schemes to recover the missing parts. However, as the number of missing entries increases, factorization schemes may overfit the model because of incorrectly predefined ranks, while completion schemes may fail to interpret the model factors. In this paper, we introduce a novel concept: complete the missing entries and simultaneously capture the underlying model structure. To this end, we propose a method called simultaneous tensor decomposition and completion (STDC) that combines a rank minimization technique with Tucker model decomposition. Moreover, as the model structure is implicitly included in the Tucker model, we use factor priors, which are usually known a priori in real-world tensor objects, to characterize the underlying joint-manifold drawn from the model factors. By exploiting this auxiliary information, our method leverages two classic schemes and accurately estimates the model factors and missing entries. We conducted experiments to empirically verify the convergence of our algorithm on synthetic data and evaluate its effectiveness on various kinds of real-world data. The results demonstrate the efficacy of the proposed method and its potential usage in tensor-based applications. It also outperforms state-of-the-art methods on multilinear model analysis and visual data completion tasks.


A Shipping Container-Based Sterile Processing Unit for Low Resources Settings.
Deficiencies in the sterile processing of medical instruments contribute to poor outcomes for patients, such as surgical site infections, longer hospital stays, and deaths. In low resources settings, such as some rural and semi-rural areas and secondary and tertiary cities of developing countries, deficiencies in sterile processing are accentuated due to the lack of access to sterilization equipment, improperly maintained and malfunctioning equipment, lack of power to operate equipment, poor protocols, and inadequate quality control over inventory. Inspired by our sterile processing fieldwork at a district hospital in Sierra Leone in 2013, we built an autonomous, shipping-container-based sterile processing unit to address these deficiencies. The sterile processing unit, dubbed "the sterile box," is a full suite capable of handling instruments from the moment they leave the operating room to the point they are sterile and ready to be reused for the next surgery. The sterile processing unit is self-sufficient in power and water and features an intake for contaminated instruments, decontamination, sterilization via non-electric steam sterilizers, and secure inventory storage. To validate efficacy, we ran tests of decontamination and sterilization performance. Results of 61 trials validate convincingly that our sterile processing unit achieves satisfactory outcomes for decontamination and sterilization and as such holds promise to support healthcare facilities in low resources settings.


Effect of N-terminal iodination on the biological, immunological and receptor binding properties of secretion. A role for the alpha-amino group of histidine in stabilizing hormone-receptor interactions.
Both radiotrace labeled and high specific activity 125I-labeled derivatives of secretin were prepared by direction iodination of the histidyl residue with chloramine T [( 125I]secretin) and by conjugation of a preiodinated Bolton-Hunter group (iodo-3-(4-hydroxyphenyl)propionate) to the free alpha-amino group at the N-terminus [( 125I]BH-secretin). Following purification, the biological, immunological and receptor binding properties of both secretin derivatives were compared. [125I]secretin and [125I]BH-secretin were equally effective in a sensitive radioimmunoassay that detected secretin and secretin (5-27) but not CCK-8, VIP and glucagon. Although both derivatives retained 60% of the biological potency of secretin as measured by cAMP accumulation in pancreatic acinar cells, only the directly iodinated peptide [( 125I]secretin) could be used to characterize specific binding sites on rat pancreatic membranes. The N-terminal blocked derivative [( 125I]BH-secretin) in contrast dissociated rapidly from pancreatic membranes reflecting an unstable hormone-receptor complex. These results suggest that a free alpha-amino group at the N-terminus may be essential for an optimal interaction of secretin with its pancreatic receptor.


Argonaute 2 is up-regulated in tissues of urothelial carcinoma of bladder.
Argonaute 2 proteins (Ago2) have been demonstrated to be widely expressed and involved in post-transcriptional gene silencing and play key roles in carcinogenesis. However, its expression profile and prognostic value in urothelial carcinoma of the bladder (UCB) have not been investigated. Real-time quantitative PCR (qRT-PCR) and Western blot were used to explore Ago2 expression in UCBs and normal bladder tissues. Moreover immunohistochemistry (ICH) was used to detect the expression of Ago2 in UCBs. Spearman's rank correlation, Kaplan-Meier plots and Cox proportional hazards regression model were used to analyze the data. Up-regulated expression of Ago2 mRNA and protein was observed in the majority of UCBs by qRT-PCR and Western blot when compared with their paired normal bladder tissues. Clinic pathological analysis was showed a significant correlation existed between the higher expression of Ago2 protein with the Histological grade, lymph node metastasis and Distant metastasis (P<0.05); Survival analysis by Kaplan-Meier survival curve and log-rank test demonstrated that elevated Ago2 expression in cancer tissue predicted poorer overall survival (OS) compared with group in lower expression (62.2% VS 86.3%, P<0.05). Notably, multivariate analyses by Cox's proportional hazard model revealed that expression of Ago2 was an independent prognostic factor in UCB. These results suggest that the aberrant expression of Ago2 in human UCB is possibly involved with tumorigenesis and development, and the Ago2 protein could act as a potential biomarker for prognosis assessment of bladder cancer. Further studies on the cellular functions of Ago2 need to address these issues.


Prostaglandin E2 Induces Expression of Mineralization Genes by Undifferentiated Dental Pulp Cells.
Prostaglandin E2 (PGE2) is a lipid mediator usually released during inflammation. This study aimed to investigate the potential of soluble or microsphere-loaded PGE2 on inducing differentiation of dental pulp stem cells. PGE2-loaded microspheres (MS) were prepared using an oil-in-water emulsion solvent extraction-evaporation process and were characterized. Mouse dental pulp stem cells (OD-21) were stimulated with soluble or PGE2-loaded MS (0.01 and 0.1 µM). Cell viability was determined by MTT colorimetric assay. Ibsp, Bmp2 and Runx2 expression was measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) after 3, 6, and 24 h. The results showed that the soluble PGE2 reduced dental pulp stem cells viability after 24 h of stimulation whereas PGE2-loaded MS did not. Soluble PGE2 up-regulated Ibsp and Bmp2 at 3 h, differently from PGE2-loaded MS. On the other hand, PGE2-MS induced Bmp2 and Runx2 at 6 h and Ibsp at 24 h. In conclusion, our in vitro results show that PGE2, soluble or loaded in MS are not cytotoxic and modulateIbsp,Bmp2, andRunx2gene expression in cultured OD-21 cells.


Telephone interviews vs. workstation sessions for acquiring quality of life data.
Patient quality of life data can be acquired in a variety of ways, including over the telephone and through computerized questionnaires. However, if the method of collection produces different results, medical decisions regarding appropriate and cost-effective care may be influenced by collection method. We conducted an experiment where subjects had two quality of life measures, the time trade-off and rating scale utilities, assessed both in telephone interivews and via computer touchscreens. The order of telephone and touchscreen was randomized. We found that rating scale utilities were similar whether obtained via the telephone or via touchscreen regardless of which was done first. However, patients who had their time trade-off utilities assessed over the telephone first did not provide as consistent responses as those elicited first via touchscreen (p = 0.01). Caution is suggested when considering eliciting time trade-off over the telephone with subjects who have not had time trade-off elicited previously.


Positive impact of preventative chemotherapy during a national helminth control program: Perception and KAP.
Helminth control at the national level is currently based on mass drug administration (MDA) programs. Perception of the MDA programs for helminth control by the affected populations influences compliance and future designs of the programs. We determined the perception of Zimbabwe's National Helminth Control Program (2012-2017) with a specific focus on schistosomiasis in the school children treated with praziquantel, schoolteachers and village health workers (VHW). The study enrolled 409 children from Grades 6 and 7 who had the full benefit of the 6 years of MDA from 2012 to 2017. Thirty-six schoolteachers and 22 VHW serving the schools were also recruited. A structured questionnaire developed in English, translated into the local language Shona, and validated prior to the study was administered to the children and the adults. The questions focused on the perceived impact on health, school attendance and performance and Knowledge Attitudes and Practice (KAP) among the school children. Data were captured electronically on android platforms using the Open Data Kit. Overall, 84% of the children responded that their awareness of schistosomiasis (transmission, disease, treatment and infection avoidance) had improved because of participating in the MDAs. Of the 151 children self-diagnosed with schistosomiasis, 74% reported that their health had improved following treatment with praziquantel. This included resolution of haematuria, painful urination, sore stomach, tiredness and falling asleep during class lessons. The children and teachers reported improvements in both pupil school attendance and performance at school while the VHW and teachers reported an increase in health-seeking behaviour amongst the school children for schistosomiasis treatment in-between MDAs. The majority of VHW (96%) reported improvement in handwashing behaviour, schistosomiasis awareness (96%) and treatment uptake (91%) within the communities where the school children belonged. However, only 59% of the VHW reported improvement in toilet use while only 50% of the VHW reported improvement in clean water use within their communities. This study indicated that the surveyed children perceived the MDA program had improved their health, school attendance, school performance and awareness of schistosomiasis. The VHW also perceived that the MDA program had improved the community KAP.


Geometry and mechanics of thin growing bilayers.
We investigate how thin sheets of arbitrary shapes morph under the isotropic in-plane expansion of their top surface, which may represent several stimuli such as nonuniform heating, local swelling and differential growth. Inspired by geometry, an analytical model is presented that rationalizes how the shape of the disk influences morphing, from the initial spherical bending to the final isometric limit. We introduce a new measure of slenderness that describes a sheet in terms of both thickness and plate shape. We find that the mean curvature of the isometric state is three fourths the natural curvature, which we verify by numerics and experiments. We finally investigate the emergence of a preferred direction of bending in the isometric state, guided by numerical analyses. The scalability of our model suggests that it is suitable to describe the morphing of sheets spanning several orders of magnitude.


Mixtures of similarly acting compounds in Daphnia magna: from gene to metabolite and beyond.
Daphnia are an important and widely studied model species in ecological and toxicological studies throughout the world and an official (OECD) recommended test organism. Their small size, wide distribution and easy growth conditions make this organism ideal for functional genomics based studies, including metabolic profiling and transcriptomics. In this study we used an integrated systems approach in which transcriptomic, metabolomic and energetic responses of juvenile (4days old) daphnids were evaluated in response to exposure to two poly aromatic hydrocarbons (pyrene and fluoranthene) and binary mixtures thereof. In addition, these responses were linked to responses measured during chronic experiments (21days) assessing survival, growth and reproductive traits. Custom Daphnia magna microarrays were used to assess transcriptomic changes. Hierarchical cluster analysis did not result in a clear distinction between the single compounds suggesting similar molecular modes of action. Cluster analysis with both the single compounds and the binary mixture treatments resulted in a separation of treatments based on differences in toxic ratios rather than component differences. Changes in the metabolic profiles of the organisms were investigated using Nuclear Magnetic Resonance Spectroscopy and Gas and Liquid Chromatography Mass Spectrometry. These multivariate metabolomic datasets were analyzed with Principal Components Analysis and Partial Least Squares Discriminant Analysis. The major metabolite changes responsible for the differences observed indicated a disturbance in aminosugar metabolism in all cases. The study demonstrates the potential of 'omics' to provide screening tools for monitoring of the freshwater environment--in invertebrate species--which is reasonably rapid, cost-effective and has the potential to greatly increase the amount of information obtained from aquatic toxicology testing.


The mycotoxin patulin: An updated short review on occurrence, toxicity and analytical challenges.
Patulin (PAT) is a common mycotoxin in fruit products, especially in apples and apple-based products. The European Commission has set maximum levels for PAT in food. Nevertheless, worrying PAT levels were recently recorded in diverse foods across the world. Therefore, a worldwide follow-up of PAT-levels in foods should be considered. Because of PAT's high probability in food products, the toxicological implications for humans need to be addressed as well. Recent studies proved adverse health effects of PAT, such as hepatotoxicity, gastrointestinal alterations and inmunotoxicity. In comparison to the toxicity of other mycotoxins such as ochratoxin A, PAT's immunotoxicity can be even more outspoken destructive. In addition, PAT is a low-molecular-weight and highly polar molecule, resulting in many analytical challenges for its detection. As the analytical techniques are continuously improving, PAT determination in multi-mycotoxin analysis has advanced using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) during the last year. Finally, the presence and toxicity of PAT requires a biomarker method to assess its exposure among the population. To date, however, there is no information regarding PAT biomarkers in biological samples. This short review highlights the PAT-occurrence profile, toxicological discoveries and analytical challenges of 2014 until to date.


Coalescence-Induced Droplet Jumping for Electro-Thermal Sensing.
Jumping droplet condensation, whereby microdroplets (ca. 1-100 μm) coalescing on suitably designed superhydrophobic surfaces jump away from the surface, has recently been shown to have a 10× heat transfer enhancement compared to filmwise condensing surfaces. However, accurate measurements of the condensation heat flux remain a challenge due to the need for low supersaturations (<1.1) to avoid flooding. The low corresponding heat fluxes (<5 W/cm2) can result in temperature noise that exceeds the resolution of the measurement devices. Furthermore, difficulties in electro-thermal measurements such as droplet and surface electrostatic charge arise in applications where direct access to the condensing surface, such as in isolated chambers and small integrated devices, is not possible. Here, we present an optical technique that can determine the experimental electro-thermal parameters of the jumping droplet condensation process with high fidelity through the analysis of jumping droplet trajectories. To measure the heat flux, we observed the experimental trajectories of condensate droplets on superhydrophobic nanostructures and simultaneously matched them in space and time with simulated trajectories using the droplet dynamic equations of motion. Two independent approaches yielded mean heat fluxes of approximately 0.13 W/cm2 with standard deviations ranging from 0.047 to 0.095 W/cm2, a 79% reduction in error when compared with classical energy balance-based heat flux measurements. In addition, we analyzed the trajectories of electrostatically interacting droplets during flight and fitted the simulated and experimental results to achieve spatial and temporal agreement. The effect of image charges on a jumping droplet as it approaches the surface was analyzed, and the observed acceleration has been numerically quantified. Our work presents a sensing methodology of electro-thermal parameters governing jumping droplet condensation.


Potential role of the peroxynitrate-poly(ADP-ribose) synthetase pathway in a rat model of severe hemorrhagic shock.
Peroxynitrite (a potent oxidant produced by nitric oxide and superoxide) and hydroxyl radical, reactive oxidants produced during hemorrhagic shock, are potent triggers of DNA single-strand breakage. DNA injury triggers the activation of the nuclear enzyme poly(ADP-ribose) synthetase (PARS), which contributes to cellular injury. Hemorrhagic shock is associated with early vasomotor paralysis as well as with early derangements in the cellular metabolic status. Here we have tested whether activation of PARS contributes to the vasodilatation and early mortality in a rat model of severe hemorrhagic shock. In anesthetized rats hemorrhaged to a mean arterial blood pressure of 35 mmHg, pretreatment with the PARS inhibitor 5-iodo-6-amino-1,2-benzopyrone significantly improved survival rate. Furthermore, an inhibitor of nitric oxide biosynthesis (NG-methyl-L-arginine) and the cell-permeable superoxide dismutase mimetic Mn(III)tetrakis(4-benzoic acid) porphyrin also offered a significant protection in terms of hypotension and acute mortality. However, the selective inhibitor of the inducible nitric oxide synthase, mercaptoethylguanidine, failed to affect blood pressure or mortality. The present data suggest that PARS activation plays a role in the pathophysiology of hemorrhagic shock. Early production of peroxynitrite (produced by nitric oxide from constitutive nitric oxide synthase) and hydroxyl radical may induce DNA single-strand breakage, which is the immediate trigger of PARS activation.


Use of Mental Health Apps by Patients With Breast Cancer in the United States: Pilot Pre-Post Study.
Nearly half of the patients with breast cancer experience clinically significant mental distress within the first year of receiving their cancer diagnosis. There is an urgent need to identify scalable and cost-efficient ways of delivering empirically supported mental health interventions to patients with breast cancer. The aim of this study was to evaluate the feasibility of in-clinic recruitment for a mobile phone app study and to evaluate the usability and preliminary impact of a suite of mental health apps (IntelliCare) with phone coaching on psychosocial distress symptoms in patients recently diagnosed with breast cancer. This pilot study adopted a within-subject, 7-week pre-post study design. A total of 40 patients with breast cancer were recruited at a US National Cancer Institute-designated clinical cancer center. Self-reported distress (Patient Health Questionnaire-4) and mood symptoms (Patient-Reported Outcomes Measurement Information System depression and anxiety scales) were assessed at baseline and postintervention. App usability was assessed at postintervention. The minimum recruitment threshold was met. There was a significant decrease in general distress symptoms, as well as symptoms of depression and anxiety, from baseline to postintervention. Overall, participants reported high levels of ease of app use and learning. Scores for app usefulness and satisfaction were reinforced by some qualitative feedback suggesting that tailoring the apps more for patients with breast cancer could enhance engagement. There is a dire need for scalable, supportive interventions in cancer. The results from this study inform how scalable mobile phone-delivered programs with additional phone support can be used to support patients with breast cancer. RR2-10.2196/11452.


Effect of ultrasound on development. Part 2: Studies in mammalian species and overview.
This report reviews and establishes patterns from the literature on experimental ultrasound exposures of inframammalian embryos and prenatal laboratory mammals. Exposure to ultrasound can produce adverse affects on development, and the sensitivity and responses of the test systems vary with the stage of development. Generally, however, deleterious effects are demonstrable only with exposure parameters that far exceed those used in clinical practice, and the few reports of mammalian embryotoxicity under clinically relevant exposure conditions have not been repeatable. Although it may be impossible to unequivocally establish absolute safety, there is a need for further studies to evaluate subtle and delayed indicators of developmental effects, potential mechanisms, and to attempt to estimate threshold exposure conditions.


Effects of Remineralizing Agents Based on Calcium Phosphate, Sodium Phosphate, or Sodium Fluoride on Eroded Cervical Dentin.
To evaluate the effect of remineralizing agents on collagen matrix pattern, precipitate formation, and dentinal tubule obliteration in eroded cervical dentin. One hundred bovine cervical dentin specimens were previously eroded (0.6% hydrochloric acid, pH 2.3, 5 minutes) and then randomized into five groups (n=20): G1, control (without treatment); G2, Desensibilize Nano P (FGM); G3, MI Paste Plus (Recaldent); G4, Regenerate (NR-5); and G5, Desensibilize KF 2% (FGM). These treatments were applied in four sessions with 7-day intervals. During this period, the samples were subjected to an erosive challenge with orange juice (pH 3.8, 5 minutes). The specimens were analyzed by polarized light microscopy with picrosirius red staining, scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX). The G3 showed a higher concentration of type I collagen than G2 and G5 (p<0.05). The G3 showed greater formation of surface precipitates than that of G1 and G5 (p<0.05). In addition, G4 and G5 showed a greater number of open dentinal tubules than that of G3 (p<0.05). Calcium phosphate-based remineralizing agents have shown to be a promising alternative treatment for preventing deleterious effects on the eroded dentin collagen matrix. In addition, they promoted precipitate formation and dentinal tubule obliteration on the eroded dentin.


Ankyrin-B regulates Kir6.2 membrane expression and function in heart.
Ankyrin polypeptides are critical for normal membrane protein expression in diverse cell types, including neurons, myocytes, epithelia, and erythrocytes. Ankyrin dysfunction results in defects in membrane expression of ankyrin-binding partners (including ion channels, transporters, and cell adhesion molecules), resulting in aberrant cellular function and disease. Here, we identify a new role for ankyrin-B in cardiac cell biology. We demonstrate that cardiac sarcolemmal K(ATP) channels directly associate with ankyrin-B in heart via the K(ATP) channel alpha-subunit Kir6.2. We demonstrate that primary myocytes lacking ankyrin-B display defects in Kir6.2 protein expression, membrane expression, and function. Moreover, we demonstrate a secondary role for ankyrin-B in regulating K(ATP) channel gating. Finally, we demonstrate that ankyrin-B forms a membrane complex with K(ATP) channels and the cardiac Na/K-ATPase, a second key membrane transporter involved in the cardiac ischemia response. Collectively, our new findings define a new role for cardiac ankyrin polypeptides in regulation of ion channel membrane expression in heart.


First person account: schizophrenia and motherhood.
With this article I want to give insight into what having children and dealing with schizophrenia is like. I go from a young woman wanting to experience motherhood, to being diagnosed with schizophrenia, to marrying and having two children. It is not easy dealing with schizophrenia and mothering, but the rewards are wonderful. In my case, I had one very serious schizophrenia episode, and I became homeless and my children went to live with their father. I eventually got better and reunited with my children, but our lives had changed. In this article I describe the adjustments we all had to make, the heartaches, the struggles, and finally the love that has prevailed. Today, especially with the newer medications, more persons suffering with mental illness are marrying, and the possibility of starting families is very real. Perhaps a young person with a diagnosis of mental illness will read my factual account, and it will help her to make a decision that will change the rest of her life. In closing the article, I raise the question, "If I had to do it over again, would I?" I also share the reason for my decision.


[What is a medication guide suits patients' needs?].
In order for a patient to read a medication guide and develop appropriate behavior regarding use of the medication, the guide should suit patients' needs. In medical care, the primary needs of patients are preventing/curing disease and/or relieving symptoms. Certainly, patients would like knowledge about what can be expected after taking a medication. However, current "Drug Guides for Patients" are based on drug labeling, which is essentially a medically sophisticated instruction manual for medical professionals who have existing knowledge about the medical treatment of the disease. Thus, there seems to be a gap in patients' needs and the contents of existing drug guides. Consequently, this disconnect may be part of the reason Drug Guides for Patients have been underused. If a patient treatment guide, which gives an overview of the disease and possible treatment strategies, is provided in conjunction with a drug guide, this combination may be useful for satisfying patients' needs. In addition, patients generally prefer detailed drug information. Consistently, surveys have revealed that many patients would like to get more information about prescribed medicine than what is frequently provided in medical practice. Furthermore, one survey reported that detailed information about possible side effects resulted in improved compliance. The need to provide patients with drug information can be considered from three points of view: patients' rights, best decision-making by the patient, and minimizing risks. Although in daily practice doctors and pharmacists may have some difficulty providing detailed medication information that includes all possible risks, more effective ways to communicate this information to patients have been suggested.


Avian and murine LR8B and human apolipoprotein E receptor 2: differentially spliced products from corresponding genes.
Apolipoprotein E-mediated lipid metabolism in the central nervous system plays an important role in cholesterol and phospholipid homeostasis of this organ, which is separated from the circulation by the blood-brain barrier. Moreover, in late-onset familial Alzheimer disease the frequency of the apolipoprotein E4 allele is significantly increased and the apoprotein is localized to extracellular plaques, one of the histological hallmarks of this disease. Recently, two distinct novel members of the low-density lipoprotein (LDL) receptor family, with the potential to bind apolipoprotein E and preferentially expressed in brain, have been characterized from human (D. Kim et al., 1996, J. Biol. Chem. 271: 8373-8380) and chicken and mouse (S. Novak, et al., 1996, J. Biol. Chem. 271: 11732-11736). The human receptor, termed "apolipoprotein E receptor 2," is a seven ligand-binding repeat receptor harboring a unique insertion in the cytoplasmic domain of the protein. The novel receptor characterized in chicken and mouse was found to have eight binding repeats without such a cytoplasmic insertion. Despite the overall identity of more than 73%, based upon their structural differences (seven versus eight ligand-binding repeats) these receptors have been considered independent entities. However, here we demonstrate that both receptors in fact are encoded by corresponding genes and that differential splicing gives rise to structurally and possibly functionally distinct variants of this brain-specific member of the LDL receptor family.


Ruthenium complexes as nitric oxide scavengers: a potential therapeutic approach to nitric oxide-mediated diseases.
1. Ruthenium(III) reacts with nitric oxide (NO) to form stable ruthenium(II) mononitrosyls. Several Ru(III) complexes were synthesized and a study made of their ability to bind NO, in vitro and also in several biological systems following expression of the inducible isoform of nitric oxide synthase (iNOS). Here we report on the properties of two, related polyaminocarboxylate-ruthenium complexes: potassium chloro[hydrogen(ethylenedinitrilo)tetraacetato]ruthenate+ ++ (=JM1226; CAS no.14741-19-6) and aqua[hydrogen(ethylenedinitrilo)tetraacetato]ruthenium (=JM6245; CAS no.15282-93-6). 2. Binding of authentic NO by aqueous solutions of JM1226 yielded a product with an infrared (IR) spectrum characteristic of an Ru(II)-NO adduct. A compound with a similar IR spectrum was obtained after reacting JM1226 with S-nitroso-N-acetylpenicillamine (SNAP). 3. The effect of JM1226 or JM6245 on nitrite (NO2-) accumulation in cultures of macrophages (RAW 264 line) 18 h after stimulating cells with lipolysaccharide (LPS) and interferon-gamma (IFNgamma) was studied. Activation of RAW264 cells increased NO2- levels in the growth medium from (mean+/-1 s.e.mean) 4.9+/-0.5 microM to 20.9+/-0.4 microM. This was blocked by actinomycin D (10 microM) or cycloheximide (5 microM). The addition of JM1226 or JM6245 (both 100 microM) to activated RAW264 cells reduced NO2- levels to 7.6+/-0.2 microM and 8.8+/-0.6 microM, respectively. N(G)-methyl-L-arginine (L-NMMA; 250 microM) similarly reduced NO2- levels, to 6.1+/-0.2 microM. 4. The effect of JM1226 or JM6245 on NO-mediated tumour cell killing by LPS+IFNgamma-activated macrophages (RAW 264) was studied in a co-culture system, using a non-adherent murine mastocytoma (P815) line as the 'target' cell. Addition of JM1226 or JM6245 (both 100 microM) to the culture medium afforded some protection from macrophage-mediated cell killing: target cell viability increased from 54.5+/-3.3% to 93.2+/-7.1% and 80.0+/-4.6%, respectively (n=6). 5. Vasodilator responses of isolated, perfused, pre-contracted rat tail arteries elicited by bolus injections (10 microl) of SNAP were attenuated by the addition of JM1226 or JM6245 (10(-4) M) to the perfusate: the ED50 increased from 6.0 microM (Krebs only) to 1.8 mM (Krebs + JM6245) and from 7 microM (Krebs only) to 132 microM (Krebs + JM1226). Oxyhaemoglobin (5 microM) increased the ED50 value for SNAP from 8 microM to 200 microM. 6. Male Wistar rats were injected with bacterial LPS (4 mg kg(-1); i.p.) to induce endotoxaemia. JM1226 and JM6245 (both 100 microM) fully reversed the hyporesponsiveness to phenylephrine of tail arteries isolated from animals previously (24 h earlier) injected with LPS. Blood pressure recordings were made in conscious LPS-treated rats using a tail cuff apparatus. A single injection of JM1226 (100 mg kg(-1), i.p.) administered 20 h after LPS (4 mg kg(-1), i.p.) reversed the hypotension associated with endotoxaemia. 7. The results show that JM1226 and JM6245 are able to scavenge NO in biological systems and suggest a role for these compounds in novel therapeutic strategies aimed at alleviating NO-mediated disease states.


Changing etiology of sympathetic ophthalmia: A 10-year study from a tertiary referral center in Taiwan.
In this study, we describe our experience of sympathetic ophthalmia (SO) at a tertiary referral center in Taiwan. We retrospectively analyzed records of patients diagnosed with SO from January 2011 to December 2020. We collected data of 15 patients diagnosed with SO (eight males, seven females). Six patients developed SO after ocular penetrating trauma, and nine developed SO after ocular surgery, including seven with vitrectomy, one with penetrating keratoplasty, and one with cataract surgery. Penetrating ocular trauma was the primary cause of SO in the first 5 years (four of six cases), but the proportion was much lower in the last 5 years (two of nine cases). The interval between trigger events and SO ranged from 0.2 to 120 months. Nine patients received oral steroids alone, five patients received methylprednisolone pulse therapy followed by oral steroids, and immunosuppressants were added in four cases. Visual acuity of sympathetic eyes and exciting eyes improved after treatment. The initial visual acuity of sympathetic eyes in trauma related and ocular surgery related revealed no significant difference, but the final visual acuity of sympathetic eyes was better in ocular surgery-related cases than in the trauma-related cases (mean ± standard deviation, 1.01 ± 1.33 versus 0.49 ± 0.68 logarithm of the minimum angle of resolution, P = 0.021). With the increased number of eye surgeries performed in recent years, eye surgery has emerged as the dominant etiology of SO in our 10-year study. Early detection and proper treatment help to maintain vision in most of the SO patients.


Genetic associations among average annual productivity, growth traits, and stayability: a parallel between Nelore and composite beef cattle.
This study was conducted to examine the relationship among average annual productivity of the cow (PRODAM), yearling weight (YW), postweaning BW gain (PWG), scrotal circumference (SC), and stayability in the herd for at least 6 yr (STAY) of Nelore and composite beef cattle. Measurements were taken on animals born between 1980 and 2010 on 70 farms located in 7 Brazilian states. Estimates of heritability and genetic and environmental correlations were obtained by Bayesian approach with 5-trait animal models. Genetic trends were estimated by regressing means of estimated breeding values by year of birth. The heritability estimates were between 0.14 and 0.47. Estimates of genetic correlation among female traits (PRODAM and STAY) and growth traits ranged from -0.02 to 0.30. Estimates of genetic correlations ranged from 0.23 to 0.94 among growth traits indicating that selection for these traits could be successful in tropical breeding programs. Genetic correlations among all traits were favorable and simultaneous selection for growth, productivity, and stayability is therefore possible. Genetic correlation between PRODAM and STAY was 0.99 and 0.85 for Nelore and composite cattle, respectively. Therefore, PRODAM and STAY might be influenced by many of the same genes. The inclusion of PRODAM instead of STAY as a selection criterion seems to be more advantageous for tropical breeding programs because the generation interval required to obtain accurate estimates of genetic merit for PRODAM is shorter. Average annual genetic changes were greater in Nelore than in composite cattle. This was not unexpected because the breeding program of composite cattle included a large number of farms, different production environments, and genetic level of the herds and breeds. Thus, the selection process has become more difficult in this population.


Contactless Monitoring Of Human Vitals: A Study With Simultaneous Measurements Using FMCW Radar And Thermal Camera.
This paper proposes contactless monitoring of Heart Rate (HR) and Breath Rate (BR) with simultaneous measurements using Frequency Modulated Continuous Wave (FMCW) radar and thermal camera. The radar collects the body movement signals which include Random Body Movements (RBMs). Non-negative Matrix Factorization (NMF) and Wavelet analysis were used on this signal to get the accurate values of HR and BR. Similarly, with thermal imaging, nostril and forehead regions are tracked to estimate the values of BR as well as HR. We conducted an experiment with 50 subjects to find similarities in the performance of radar and thermal camera while measuring HR and BR. Simultaneously, these two methods have been validated with pulse oximeter and visual camera. From the visual camera, we can get the abdominal movements on which the BR can be ascertained whereas pulse oximeter gives us the HR. Radar signals are degraded because of large RBMs whereas thermal signals get distorted because of sudden temperature changes in the surroundings, sweating, and occlusion. We used a Signal Quality Metric (SQM) to ascertain the measurement quality of the vital signs. This SQM-based approach can further be used for sensor fusion to build a robust contactless system to monitor vital signs.Clinical relevance- Contactless and accurate measurement of HR and BR is very essential for continuous and comfortable monitoring of vitals. In this paper, we combine both FMCW radar and thermal camera so that one can complement the other in adverse scenarios on the basis of signal quality.


[Modern hygiene products impact on oral microbial, pH and mineral balance].
Several toothpastes are compared in the study: "Zhemchuzhnaya-complex protection" containing as abrasive substance finely dispersed dicalcium phosphate phosphathydrate, "Noviy zhemchug ftor" and "Zhemchug svezhaya myata" with calcium carbonate. "Zhemchuzhnaya-complex protection" and "Noviy zhemchug ftor" both contain sodium monophosphate as active substance. Impact of these toothpastes on oral microbial, pH and mineral balance was assessed in the study.


[The nature of "hydropic degeneration" of hepatocytes in severe viral hepatitis].
Specimens were fixed with a mixed fluid of osmium acid potassium ferrocyanide, and embedded with Epon--812. Totally, biopsies from 21 cases of severe chronic viral hepatitis were investigated. When hydropic degeneration of hepatocytes occurred, the clear field in cytoplasm were actually places for accumulating massive glycogen. The granules around the nuclei were convergent mitochondria, endoplasm reticulum and other organelles. Glycogen was best shown in this experiment because of adoption of the mixed fluid as the fixative, which successfully prevented the loss of glycogen during dehydration and the process of uranyl acetate staining. In our opinion this change in hepatocytes might better be termed "glycogen degeneration". It is probably related to infusion of large amount of glucose fluid or due to damage of certain glycogenolytic enzymes in the liver.


The delta subunit of AP-3 is required for efficient transport of VSV-G from the trans-Golgi network to the cell surface.
Vesicular stomatitis virus glycoprotein (VSV-G) is a transmembrane protein that functions as the surface coat of enveloped viral particles. We report the surprising result that VSV-G uses the tyrosine-based di-acidic motif (-YTDIE-) found in its cytoplasmic tail to recruit adaptor protein complex 3 for export from the trans-Golgi network. The same sorting code is used to recruit coat complex II to direct efficient transport from the endoplasmic reticulum to the Golgi apparatus. These results demonstrate that a single sorting sequence can interact with sequential coat machineries to direct transport through the secretory pathway. We propose that use of this compact sorting domain reflects a need for both efficient endoplasmic reticulum export and concentration of VSV-G into specialized post-trans-Golgi network secretory-lysosome type transport containers to facilitate formation of viral coats at the cell surface.


The effect of intravenous tranexamic acid on blood loss and early post-operative pain in total shoulder arthroplasty.
The purpose of the present study was to evaluate the impact of intravenous tranexamic acid on the reduction of blood loss, transfusion rate, and early post-operative clinical outcome in total shoulder arthroplasty. A randomised, placebo-controlled trial which included 54 patients undergoing unilateral primary stemless anatomical or stemmed reverse total shoulder arthroplasty was undertaken. Patients received either 100 ml saline (placebo, n = 27), or 100 ml saline together with 1000 mg of tranexamic acid (TXA, n = 27) intravenously prior to skin incision and during wound closure. Peri-operative blood loss via an intra-articular drain was recorded and total blood loss was calculated. The post-operative transfusion rate was documented. Assessment of early clinical parameters included the visual analogue scale for pain (VAS), documentation of haematoma formation and adverse events. Mean peri-operative blood drainage (placebo: 170 ml versus TXA: 50 ml, p = 0.001) and calculated mean total blood loss (placebo: 1248.2 ml versus TXA: 871.0 ml, p = 0.009) were significantly lower in the TXA group. No transfusions were necessary during the study period in either group. Mean VAS for pain significantly decreased from pre-operative (VAS 7) to the early post-operative period (VAS 1.7, p < 0.001). Significant differences regarding mean post-operative pain between placebo (VAS 2.0) and TXA (VAS 1.3) were detected (p = 0.05). The occurrence of haematomas was significantly more frequent in the placebo (59.3%, n = 16) than in the TXA group (25.9%, n = 6, p = 0.027). Whereas only mild haematomas developed in the TXA group, in the placebo group a total of 22.2% (n = 6) developed either moderate or severe haematomas. No adverse events associated with administration of TXA occurred. Intravenous administration of TXA successfully reduced mean peri-operative blood drainage, total estimated blood loss, pain during the first post-operative days, and haematoma formation in total shoulder arthroplasty. Cite this article: Bone Joint J 2017;99-B:1073-9.


Molecular characterization of a glycosylphosphatidylinositol-linked ADP-ribosyltransferase from lymphocytes.
Mono ADP-ribosyltransferases catalyze the transfer of the ADP-ribose moiety of nicotinamide adenine dinucleotide (NAD) to proteins. It was reported by Wang et al (J Immunol 153:4048, 1994) that incubation of mouse cytotoxic T lymphocytes (CTL) with NAD resulted in the ADP-ribosylation of membrane proteins and inhibition of cell proliferation and cytotoxicity. Treatment of CTL with phosphatidylinositol-specific phospholipase C (PI-PLC) before incubation with NAD prevented the inhibitory effects of NAD on the cells, consistent with the removal of a glycosylphosphatidylinositol (GPI)-anchored ADP-ribosyltransferase on the lymphocyte surface. We have identified and cloned a GPI-linked ADP-ribosyltransferase from Yac-1 mouse T-cell lymphoma cells. The deduced amino acid sequence of the Yac-1 transferase was 70% and 41% identical to those of the rabbit skeletal muscle and chicken heterophil, respectively. It contained three noncontiguous sequences similar to those found in several of the bacterial toxin and vertebrate ADP-ribosyltransferases. Based on crystallography of the bacterial toxins, these regions are believed to form, in part, the catalytic site consistent with a common mechanism for the ADP-ribose transfer reaction. In rat mammary adenocarcinoma (NMU) cells transformed with the Yac-1 transferase cDNA, transferase activity was present on the cell surface and was released into the medium by treatment of cells with PI-PLC. Thus, we have cloned a novel gene that has properties identical to the transferase detected in CTL, and may be involved in the NAD-dependent regulation of proliferation and cytotoxicity.


Level of AIDS and HIV knowledge and sexual practices among sexually transmitted disease patients in China.
Little is known about sex practices that increase the risk of contracting HIV infection or the level of HIV and AIDS knowledge among sexually transmitted disease (STD) patients in China. To describe AIDS and HIV knowledge, sexual practices, and factors associated with never using condoms among patients at an STD clinic in Jinan, China. Clinic patients (n = 498) were randomly sampled to answer AIDS and HIV knowledge questions and to report sexual practices, including condom use. Patients had low levels of AIDS and HIV knowledge and engaged in high-risk sex behaviors. The majority of patients reported having had multiple sex partners. When having sex, few men and no women reported always using condoms. Gender, age, residence, AIDS and HIV knowledge, and having multiple sex partners were significantly associated with never using condoms. STD clinic patients report having engaged in high-risk sex behaviors. More research is needed to better understand the factors relevant to developing risk-reduction interventions for these patients in China.


Naturally processed and presented epitopes of the islet cell autoantigen IA-2 eluted from HLA-DR4.
During immune responses, antigen-presenting cells (APCs) process antigens and present peptide epitopes complexed with human leukocyte antigen (HLA) molecules. CD4 cells recognize these naturally processed and presented epitopes (NPPEs) bound to HLA class II molecules. Epitope identification is important for developing diagnostic and therapeutic tools for immune-mediated diseases and providing insight into their etiology, but current approaches overlook effects of natural processing on epitope selection. We have developed a technique to identify NPPEs using mass spectrometry (MS) after antigen is targeted onto APCs using a lectin-based antigen delivery system (ADS). We applied the technique to identify NPPEs of the intracellular domain of the type 1 diabetes mellitus-associated (type 1 DM-associated) autoantigen insulinoma-associated-2 (IA-2ic), presented by HLA-DR4 (0401). IA-2ic-derived NPPEs eluted from HLA-DR4 constitute 6 sets of peptides nested around distinct core regions. Synthetic peptides based on these regions bind HLA-DR4 and elicit primary T-cell proliferation frequently in HLA-DR4-positive type 1 DM patients, but rarely in non-HLA-DR4 patients, and in none of the HLA-DR4 nondiabetic controls we tested. This flexible, direct approach identifies an HLA allele-specific map of NPPEs for any antigen, presented by any HLA class II molecule. This method should enable a greater understanding of epitope selection and lead to the generation of sensitive and specific reagents for detecting autoreactive T cells.


The control of ascorbic acid synthesis and turnover in pea seedlings.
The rate of ascorbate synthesis and turnover in pea seedling embryonic axes was investigated in relation to its pool size. Ascorbate accumulated in embryonic axes of germinating pea seeds which has been supplied with ascorbate. Incorporation of [U-14C]glucose into ascorbate after a 2 h labelling period was reduced by ascorbate loading for 3 h and 20 h, providing evidence that ascorbate biosynthesis is inhibited by endogenous ascorbate. Ascorbate turnover was estimated by following the metabolism of [1-14C]ascorbate over 2 h after ascorbate loading and by the rate of decrease of the ascorbate pool size after ascorbate loading. Ascorbate turnover rate, determined by [1-14C]ascorbate metabolism, increased as a linear function of pool size. The absolute turnover rate was higher in ascorbate-loaded embryonic axes but was always about 13% of the pool per hour. The initial rate of ascorbate turnover, estimated from the net decrease in pool size after ascorbate loading, also showed a similar turnover rate to that estimated from [1-14C]ascorbate metabolism. Ascorbate loading had no effect on ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase or glutathione reductase activity. Ascorbate oxidase activity decreased after ascorbate loading.


Mitigation of oxygen-induced retinopathy in α2β1 integrin-deficient mice.
The α2β1 integrin plays an important but complex role in angiogenesis and vasculopathies. Published GWAS studies established a correlation between genetic polymorphisms of the α2β1 integrin gene and incidence of diabetic retinopathy. Recent studies indicated that α2-null mice demonstrate superior vascularization in both the wound and diabetic microenvironments. The goal of this study was to determine whether the vasculoprotective effects of α2-integrin deficiency extended to the retina, using the oxygen-induced retinopathy (OIR) model for retinopathy of prematurity (ROP). In the OIR model, wild-type (WT) and α2-null mice were exposed to 75% oxygen for 5 days (postnatal day [P] 7 to P12) and subsequently returned to room air for 6 days (P12-P18). Retinas were collected at postnatal day 7, day 13, and day 18 and examined via hematoxylin and eosin and Lectin staining. Retinas were analyzed for retinal vascular area, neovascularization, VEGF expression, and Müller cell activation. Primary Müller cell cultures from WT and α2-null mice were isolated and analyzed for hypoxia-induced VEGF-A expression. In the retina, the α2β1 integrin was minimally expressed in endothelial cells and strongly expressed in activated Müller cells. Isolated α2-null primary Müller cells demonstrated decreased hypoxia-induced VEGF-A expression. In the OIR model, α2-null mice displayed reduced hyperoxia-induced vaso-attenuation, reduced pathological retinal neovascularization, and decreased VEGF expression as compared to WT counterparts. Our data suggest that the α2β1 integrin contributes to the pathogenesis of retinopathy. We describe a newly identified role for α2β1 integrin in mediating hypoxia-induced Müller cell VEGF-A production.


Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein.
In order to deliver low-cost viral capsomeres from a large amount of soluble viral VP6 protein from human rotavirus, we developed and optimized a biotechnological platform in Escherichia coli. Specifically, three different expression protocols were compared, differing in their genetic constructs, ie, a simple native histidine-tagged VP6 sequence, VP6 fused to thioredoxin, and VP6 obtained with the newly described small ubiquitin-like modifier (SUMO) fusion system. Our results demonstrate that the histidine-tagged protein does not escape the accumulation in the inclusion bodies, and that SUMO is largely superior to the thioredoxin-fusion tag in enhancing the expression and solubility of VP6 protein. Moreover, the VP6 protein produced according to the SUMO fusion tag displays well-known assembly properties, as observed in both transmission electron microscopy and atomic force microscopy images, giving rise to either VP6 trimers, 60 nm spherical virus-like particles, or nanotubes a few microns long. This different quaternary organization of VP6 shows a higher level of immunogenicity for the elongated structures with respect to the spheres or the protein trimers. Therefore, the expression and purification strategy presented here - providing a large amount of the viral capsid protein in the native form with relatively simple, rapid, and economical procedures - opens a new route toward large-scale production of a more efficient antigenic compound to be used as a vaccination tool or as an adjuvant, and also represents a top-quality biomaterial to be further modified for biotechnological purposes.


Metallothionein is a crucial protective factor against Helicobacter pylori-induced gastric erosive lesions in a mouse model.
Infection with the gastric pathogen Helicobacter pylori (H. pylori) causes chronic gastritis, peptic ulcer, and gastric adenocarcinoma. These diseases are associated with production of reactive oxygen species (ROS) from infiltrated macrophages and neutrophiles in inflammatory sites. Metallothionein (MT) is a low-molecular-weight, cysteine-rich protein that can act not only as a metal-binding protein, but also as a ROS scavenger. In the present study, we examined the role of MT in the protection against H. pylori-induced gastric injury using MT-null mice. Female MT-null and wild-type mice were challenged with H. pylori SS1 strain, and then histological changes were evaluated with the updated Sydney grading system at 17 and 21 wk after challenge. Although the colonization efficiency of H. pylori was essentially the same for MT-null and wild-type mice, the scores of activity of inflammatory cells were significantly higher in MT-null mice than in wild-type mice at 17 wk after challenge. Histopathological examination revealed erosive lesions accompanied by infiltration of inflammatory cells in the infected MT-null mice but not in wild-type mice. Furthermore, activation of NF-kappaB and expression of NF-kappaB-mediated chemokines such as macrophage inflammatory protein-1alpha and monocytes chemoattractant protein-1 in gastric cells were markedly higher in MT-null mice than in wild-type mice. These results suggest that MT in the gastric mucosa might play an important role in the protection against H. pylori-induced gastric ulceration.


Enrichment of prostate cancer stem-like cells from human prostate cancer cell lines by culture in serum-free medium and chemoradiotherapy.
The discovery of rare subpopulations of cancer stem cells (CSCs) has created a new focus in cancer research. As CSCs demonstrate resistance to chemoradiation therapy relative to other cancer cells, this allows the enrichment of CSC populations by killing apoptosis-susceptible cancer cells. In this study, three commonly used human prostate cancer (PCa) cell lines (DU145, PC-3 and LNCaP) were examined for their expression of the putative stem cell markers CD133 and CD44 via flow cytometric analysis. Under normal culture conditions, CD133(+)/CD44(+) cells were only present in the DU145 cell line, and comprised only a minor percentage (0.1% ± 0.01%) of the total population. However, the proportion of these CD133(+)/CD44(+) prostate CSCs could be increased in these cell lines via culture in serum-free medium (SFM), or through chemotherapy or radiotherapy. Indeed, after culture in SFM, the proportion of CD133(+)/CD44(+) cells in DU145 and PC-3 had increased to 10.3% and 3.0%, respectively. Moreover, the proportion had increased to 9.8% enriched by chemotherapy and 3.5% by radiotherapy in DU145. Colony-formation tests, cell invasion assays, and tumor xenografts in BALB/c nude mice were used to evaluate the stem cell properties of CD133(+)/CD44(+) PCa cells that were isolated via fluorescence-activated cell sorting (FACS). CD133(+)/CD44(+) cells had an enhanced colony-formation capability and invasive ability in vitro, and displayed greater tumorigenic properties in vivo. These results demonstrate the presence of CD133(+)/CD44(+) prostate CSCs in established PCa cell lines and that populations of these cells can be enriched by culture in SFM or chemoradiotherapy. Finding novel therapies to override chemoradiation resistance in the prostate CSCs is the key to improve long-term results in PCa management.