The Effects of Source Water Quality on Drinking Water Treatment Costs: A Review and Synthesis of Empirical Literature.
Watershed protection, and associated in situ water quality improvements, has received considerable attention as a means for mitigating health risks and avoiding expenditures at drinking water treatment plants (DWTPs). This study reviews the literature linking source water quality to DWTP expenditures. For each study, we report information on the modeling approach, data structure, definition of treatment costs and water quality, and statistical methods. We then extract elasticities indicating the percentage change in drinking water treatment costs resulting from a 1% change in water quality. Forty-six elasticities are obtained for various water quality parameters, such as turbidity, total organic carbon (TOC), nitrogen, sediment loading, and phosphorus loading. An additional 29 elasticities are obtained for land use classification (e.g., forest, agricultural, urban), which often proxy source water quality. Findings indicate relatively large ranges in the estimated elasticities of most parameters and land use classifications. However, average elasticities are smaller and ranges typically narrower for studies that incorporated control variables consistent with economic theory in their models. We discuss the implications of these findings for a DWTP's incentive to engage in source water protection and highlight gaps in the literature.


Mitigation of harmful chemical formation from pyrolysis of tobacco waste using CO2.
Global consumption of tobacco has been continuously increasing. This results in the considerable generation of toxic waste materials from the tobacco industry and daily life. Conventional disposal methods for them (incineration and landfilling) could be a potential hazard for releasing carcinogens and toxins into our eco-system. Accordingly, an eco-friendly disposal platform for converting tobacco waste (TW) into syngas was mainly studied in this present work. To realize this, pyrolysis of two commercial cigarette products (Marlboro and HEETS (electronic cigarette)) was done under the CO2/N2 conditions. One of the main findings from the present study was that CO2 reacted with volatile matters (VMs) obtained from the thermolysis of TW through the gas phase reactions (GPRs), which provided a strategic measure to manipulate carbon rearrangement of all pyrolysates. In particular, the GPRs expedited the carbon rearrangement of harmful chemical species, converting toxic chemicals into syngas. When the fraction of VMs in TWs increased, the GPR were more effective. Therefore, the introduced eco-friendly method using CO2-mediated thermochemical process could be beneficial for energy recovery from TWs while mitigating the formations of harmful chemical species.


Evidence of both extra- and intracellular cysteine targets of protein modification for activation of RET kinase.
By use of a specifically sulfhydryl group-reactive chemical, 1,4-butanediyl-bismethanethiosulfonate (BMTS), we studied the localization of oxidative stress-responsive target cysteines for activation of a receptor-type protein tyrosine kinase, c-RET. The chemical, which reacted with RET proteins on the cell surface for sulfhydryl-linked aggregation, induced autophosphorylation and activation of RET kinase. When extracellular domain-deleted RET mutant (RET-PTC-1) cells were exposed to BMTS, neither the molecular status nor the activity of the enzyme was affected, suggesting that the target cysteines of BMTS to which cells were exposed for reaction are located in the cysteine-rich region of the extracellular domain of RET kinase. Despite this result, the exposure of a subcellular form of c-RET or RET-PTC-1 kinase isolated by immunoprecipitation to BMTS did induce activation of the enzyme. These results suggest that cysteines in both the extracellular and the intracellular domains of RET can work as target sites of accessible BMTS and possibly other oxidative elements for structural modification and activation of RET kinase.


Hypoxia in Gliomas: Opening Therapeutical Opportunities Using a Mathematical-Based Approach.
This chapter explores the use of mathematical models as promising and powerful tools to understand the complexity of tumors and their, frequently, hypoxic environment. We focus on gliomas, which are primary brain tumors derived from glial cells, mainly astrocytes and/or oligodendrocytes. A variety of mathematical models, based on ordinary and/or partial differential equations, have been developed both at the micro and macroscopic levels. The aim here is to describe in a quantitative way key physiopathological mechanisms relevant in these types of malignancies and to suggest optimal therapeutical strategies. More specifically, we consider novel therapies targeting thromboembolic phenomena to decrease cell invasion in high grade glioma or to delay the malignant transformation in low grade gliomas. This study has been the basis of a multidisciplinary collaboration involving, among others, neuro-oncologists, radiation oncologists, pathologists, cancer biologists, surgeons and mathematicians.


Implementation of long-wavelength cut-off filters based on critical angle.
Our first attempts at the fabrication of long-wavelength infrared cut-off filters with extended transmission and rejection regions that are based on the use of the critical angle, the dispersion of refractive indices, and on thin-film interference were not very successful. The design of the filter consisted of layers placed at the interface between two high-index prisms. Using the available deposition equipment, the layers produced were porous and very rough. The pores adsorbed water vapor, which resulted in absorption. The roughness made the process of optical contacting very difficult. In this paper we describe the adjustments in the design and deposition processes that allowed us to obtain filters with a better and more stable performance.


Ambulatory foot temperature measurement: a new technique in polyneuropathy evaluation.
Complaints of abnormal foot temperature are common among patients with polyneuropathy. However, there is no published method of ambulatory foot temperature measurement to identify possible thermoregulatory disturbances in these patients. We configured a digital electronic thermometer and thermocouple to measure and record distal foot and ambient temperatures simultaneously every minute for 24 to 48 h. Sixteen patients with polyneuropathy and 5 normal subjects were studied; 12 patients with polyneuropathy and 4 normal subjects had at least 24 h of successful recording. The data obtained from these patients were consistent and easily summarized by standard statistical methods. In the others, technical difficulties produced nonphysiological readings. In the patients with polyneuropathy, changes in foot temperature mirrored ambient temperature fluctuations more closely than in normal subjects. This technique shows promise in studying temperature regulation in the feet and may provide new insights into neuropathy-associated pain and the pathogenesis of polyneuropathy.


Diagnosis-independent loss of T-cell costimulatory molecules in individuals with cytomegalovirus infection.
Major depressive disorder (MDD) is associated with physiological changes commonly observed with increasing age, such as inflammation and impaired immune function. Age-related impaired adaptive immunity is characterized by the loss of naive T-cells and the reciprocal accumulation of memory T-cells together with the loss of T-cell co-stimulatory molecules. Additionally, the presence and activity of cytomegalovirus (CMV) alters the architecture of the T-cell compartment in a manner consistent with premature aging. Because CMV is also thought to reactivate with psychological stress, this study tested whether MDD influences age-related phenotypes of T-cell populations in the context of CMV infection in young and middle-aged adults. Morning blood samples from volunteers with a DSM-IV diagnosis of MDD (n = 98, mean age(SD) = 36(10) years, 74.5% female, 57.1% CMV+) and comparison controls (n = 98, mean age(SD) = 34(10) years, 68.4% female, 51.0% CMV+) were evaluated for CMV IgG antibody status and the distribution of late differentiated (CD27-CD28-) cells within CD4+ and CD8+ T-cell subsets, i.e. naive (CCR7+CD45RA+), effector memory (EM, CCR7-CD45RA-), central memory (CM, CCR7+CD45RA-) and effector memory cells re-expressing CD45RA (EMRA, CCR7-CD45RA+). Mixed linear regression models controlling for age, sex, ethnicity and flow cytometry batch showed that CMV seropositivity was associated with a reduction in naive T-cells, expansion of EMRA T-cells, and a greater percent distribution of CD27-CD28- cells within CD4+ and CD8+ memory T-cell subsets (p's < 0.004), but there was no significant effect of MDD, nor any significant interaction between CMV and diagnosis. Unexpectedly, depressed men were less likely to be CMV+ and depressed women were more likely to be CMV+ than sex-matched controls suggesting a possible interaction between sex and MDD on CMV susceptibility, but this three-way interaction did not significantly affect the T-cell subtypes. Our findings suggest that depression in young and middle-aged adults does not prematurely advance aging of the T-cell compartment independently of CMV, but there may be significant sex-specific effects on adaptive immunity that warrant further investigation.


Gender Differences in Personal and Situational Risk Factors for Traumatic Brain Injury Among Older Adults.
To investigate differences between older men and women in Taiwan in personal and situational risk factors for sustaining a traumatic brain injury (TBI) versus soft-tissue injury (STI) due to a fall. Matched case-control study. Cases were defined as patients with a primary diagnosis of TBI due to a fall and were identified from those 60 years or older who visited the emergency department (ED) of 3 university-affiliated hospitals in 2015. Matched by the same hospital ED, gender, and time of falling, 3 controls who had no TBI and who had sustained only soft-tissue injury (STI) due to falling were selected for comparison with each case. Personal factors and situational exposures were compared between the control and case groups. In total, 96 cases and 288 controls in men and 72 cases and 216 controls in women participated in this study. Personal factors (sociodemographic and lifestyle factors, medical characteristics, and functional abilities) and situational exposures (location, activities before the fall, center-of-mass change, type of fall, falling direction, protective response, and impact during the fall). In men, after adjusting for other variables, older age (odds ratio [OR] = 1.04), regular alcohol use (OR = 2.03), an indoor fall (OR = 1.92), activity of getting in/out of bed (OR = 2.56), a fall due to dizziness (OR = 4.09), and falling backward (OR = 2.95) were independently associated with a higher odds of TBI. In women, an older age (OR = 1.03), the presence of Parkinson disease (OR = 10.4), activities of toileting (OR = 2.50), getting in/out of bed (OR = 4.90), and negotiating stairs (OR = 7.13), a fall due to dizziness (OR = 5.05), and falling backward (OR = 2.61) were independently associated with a higher odds of TBI. Our results demonstrated similarities and differences in personal and situational risk factors for fall-related TBIs versus STIs between older men and women, and gender differences should be considered when developing intervention strategies.


Mapping 123 million neonatal, infant and child deaths between 2000 and 2017.
Since 2000, many countries have achieved considerable success in improving child survival, but localized progress remains unclear. To inform efforts towards United Nations Sustainable Development Goal 3.2-to end preventable child deaths by 2030-we need consistently estimated data at the subnational level regarding child mortality rates and trends. Here we quantified, for the period 2000-2017, the subnational variation in mortality rates and number of deaths of neonates, infants and children under 5 years of age within 99 low- and middle-income countries using a geostatistical survival model. We estimated that 32% of children under 5 in these countries lived in districts that had attained rates of 25 or fewer child deaths per 1,000 live births by 2017, and that 58% of child deaths between 2000 and 2017 in these countries could have been averted in the absence of geographical inequality. This study enables the identification of high-mortality clusters, patterns of progress and geographical inequalities to inform appropriate investments and implementations that will help to improve the health of all populations.


Typing of Legionella strains isolated from environmental samples in Crete, Greece, during the period 2004-2011.
In Greece, standard tests are performed in watering and cooling systems of hotels. A total of 1,494 water samples were collected during 2004-2011 from 124 hotels from four regions in Crete (Greece). Samples were tested for the presence of Legionella spp.; 103 isolates were identified and typed by polymerase chain reaction (PCR)-sequencing and sequence-based typing (SBT) (in case of L. pneumophila sg 1). Of those, 48 belonged to various serogroups of L. pneumophila (sg 1, 2, 3, 5, 6, 8, 12, 13, and 15), 32 were characterized as L. anisa, 17 as L. taurinensis and there was a single occurrence of L. quinlivanii, L. maceachernii, and L. oakridgensis. In the case of L. pneumophila SG1, one prevalent sequence type was revealed (ST37). The variability of Legionella spp. observed questions the existence of a single ST of the L. pneumophila sg1 species and leads towards the need for a genetic level investigation of all Legionnaires' disease cases.


Identification of genes involved in low aminoglycoside-induced SOS response in Vibrio cholerae: a role for transcription stalling and Mfd helicase.
Sub-inhibitory concentrations (sub-MIC) of antibiotics play a very important role in selection and development of resistances. Unlike Escherichia coli, Vibrio cholerae induces its SOS response in presence of sub-MIC aminoglycosides. A role for oxidized guanine residues was observed, but the mechanisms of this induction remained unclear. To select for V. cholerae mutants that do not induce low aminoglycoside-mediated SOS induction, we developed a genetic screen that renders induction of SOS lethal. We identified genes involved in this pathway using two strategies, inactivation by transposition and gene overexpression. Interestingly, we obtained mutants inactivated for the expression of proteins known to destabilize the RNA polymerase complex. Reconstruction of the corresponding mutants confirmed their specific involvement in induction of SOS by low aminoglycoside concentrations. We propose that DNA lesions formed on aminoglycoside treatment are repaired through the formation of single-stranded DNA intermediates, inducing SOS. Inactivation of functions that dislodge RNA polymerase leads to prolonged stalling on these lesions, which hampers SOS induction and repair and reduces viability under antibiotic stress. The importance of these mechanisms is illustrated by a reduction of aminoglycoside sub-MIC. Our results point to a central role for transcription blocking at DNA lesions in SOS induction, so far underestimated.


Long distance dispersal and vertical gene flow in the Caribbean brooding coral Porites astreoides.
To date, most assessments of coral connectivity have emphasized long-distance horizontal dispersal of propagules from one shallow reef to another. The extent of vertical connectivity, however, remains largely understudied. Here, we used newly-developed and existing DNA microsatellite loci for the brooding coral Porites astreoides to assess patterns of horizontal and vertical connectivity in 590 colonies collected from three depth zones (≤10 m, 15-20 m and ≥25 m) at sites in Florida, Bermuda and the U.S. Virgin Islands (USVI). We also tested whether maternal transmission of algal symbionts (Symbiodinium spp.) might limit effective vertical connectivity. Overall, shallow P. astreoides exhibited high gene flow between Florida and USVI, but limited gene flow between these locations and Bermuda. In contrast, there was significant genetic differentiation by depth in Florida (Upper Keys, Lower Keys and Dry Tortugas), but not in Bermuda or USVI, despite strong patterns of depth zonation in algal symbionts at two of these locations. Together, these findings suggest that P. astreoides is effective at dispersing both horizontally and vertically despite its brooding reproductive mode and maternal transmission of algal symbionts. In addition, these findings might help explain the ecological success reported for P. astreoides in the Caribbean in recent decades.


Integration of disability-related content in nursing curricula.
Despite evidence that health care professionals, including nurses, do not perceive persons with disabilities in a positive light and fail to attend to their health care needs, it is not known how nursing programs address the topic of disability. This descriptive study was conducted to examine the extent to which schools of nursing in the United States address disability-related issues and the strategies used to integrate disability-related content in their curricula. A 27-item investigator-developed survey was sent to a national stratified random sample of 1,000 schools of nursing. Questions pertained to curricular content, specific groups of persons with disabilities, and teaching methods and resources used to teach nursing students about disability. Respondents indicated that they included some content related to disability in their curricula. Nursing textbooks were the most common source of information used. Barriers to including disability-related content were lack of time and lack of faculty interest or expertise. These findings can serve as a stimulus to increase the extent and breadth of disability-related issues in nursing curricula.


Effects of various anesthetic agents on laryngeal motion during laryngoscopy in normal dogs.
To evaluate the effects of various drugs and drug combinations conventionally used for anesthesia on arytenoid cartilage motion during laryngoscopy in normal dogs. Experimental study. Six large breed healthy dogs with no previous history of respiratory dysfunction. Each dog was randomly assigned to a different injectable anesthetic protocol once weekly for 6 weeks, then in the 7th week all dogs were anesthetized with isoflurane. Videolaryngoscopy was performed and recorded starting immediately after induction until dogs could no longer be safely restrained for endoscopy. Video was digitized and 3 still images of maximal inspiration and expiration from the first 15 seconds (induction) and the last 15 seconds (recovery) were captured and imported into an image analysis software program. The height and area of the laryngeal ostium were measured in pixels. Normalization of the glottal gap area was performed using the formula (normalized glottal gap area (NGGA)=area in pixels/height(2)). ANOVA was performed on the NGGA of images collected at inspiration and expiration during induction and recovery. Fischer's exact test was performed when significance (P<.05) was found. Within each protocol, laryngeal motion (defined as change in NGGA) at induction was not significantly different from laryngeal motion measured at recovery. Additionally, no significant differences were found in arytenoid motion immediately after induction when anesthetic protocols were compared. Arytenoid motion before recovery was significantly greater with thiopental when compared with propofol (P=.046), ketamine+diazepam (P=.0098), acepromazine+thiopental (P=.0021), and acepromazine+propofol (P=.0065). No significant difference in arytenoid motion was seen immediately after induction or before recovery when acepromazine+butorphanol+ isoflurane and thiopental were compared. We concluded that intravenous thiopental given to effect is the best choice for assessing laryngeal function in dogs. Dogs premedicated with acepromazine with or without opioids that require further anesthetic restraint for laryngoscopy should be anesthetized with isoflurane administered by mask. Misdiagnosis of laryngeal paralysis during laryngoscopy can be avoided by selecting the anesthetic regimens with the least effect on arytenoid motion.


Dural laceration occurring with burst fractures and associated laminar fractures.
The cases of sixty patients in whom a burst fracture of a thoracic or lumbar vertebral body had been treated with posterior instrumentation and arthrodesis less than two weeks after the injury were retrospectively reviewed. Thirty of the patients had an associated laminar fracture. Eleven of the thirty, all of whom had a lumbar fracture and a preoperative neurological deficit, were noted at operation to have dural laceration. In four of the patients who had dural laceration, neural elements were entrapped between the fragments from the laminar fracture. None of the remaining thirty patients who did not have a laminar fracture had dural laceration (p = 0.0002). Univariate and multivariate statistical analysis revealed no significant association of the dural laceration with the patients' age or sex, or with the radiographic characteristics of the spine. There was a significant association between dural laceration and neurological deficit (p = 0.0001). In our series, the presence of a preoperative neurological deficit in a patient who had a burst fracture and an associated laminar fracture was a sensitive (100 per cent) and specific (74 per cent) predictor of dural laceration. The presence of this fracture pattern and an associated neurological deficit also predicted a risk of dural laceration with entrapped neural elements. This information may influence decisions as to whether an anterior or a posterior surgical approach should be used in such patients.


Secretion of a mammalian chondroitinase ABC aids glial integration at PNS/CNS boundaries.
Schwann cell grafts support axonal growth following spinal cord injury, but a boundary forms between the implanted cells and host astrocytes. Axons are reluctant to exit the graft tissue in large part due to the surrounding inhibitory environment containing chondroitin sulphate proteoglycans (CSPGs). We use a lentiviral chondroitinase ABC, capable of being secreted from mammalian cells (mChABC), to examine the repercussions of CSPG digestion upon Schwann cell behaviour in vitro. We show that mChABC transduced Schwann cells robustly secrete substantial quantities of the enzyme causing large-scale CSPG digestion, facilitating the migration and adhesion of Schwann cells on inhibitory aggrecan and astrocytic substrates. Importantly, we show that secretion of the engineered enzyme can aid the intermingling of cells at the Schwann cell-astrocyte boundary, enabling growth of neurites over the putative graft/host interface. These data were echoed in vivo. This study demonstrates the profound effect of the enzyme on cellular motility, growth and migration. This provides a cellular mechanism for mChABC induced functional and behavioural recovery shown in in vivo studies. Importantly, we provide in vitro evidence that mChABC gene therapy is equally or more effective at producing these effects as a one-time application of commercially available ChABC.


Spontaneous eyeblinks during breaking continuous flash suppression are associated with increased detection times.
An eyeblink has a clear effect on low-level information processing because it temporarily occludes all visual information. Recent evidence suggests that eyeblinks can also modulate higher level processes (e.g., attentional resources), and vice versa. Despite these putative effects on different levels of information processing, eyeblinks are typically neglected in vision and in consciousness research. The main aim of this study was to investigate the timing and the effect of eyeblinks in an increasingly popular paradigm in consciousness research, namely breaking continuous flash suppression (b-CFS). Results show that participants generally refrain from blinking during a trial, that is, when they need to detect a suppressed stimulus. However, when they do blink during a trial, we observed a sharp increase in suppression time. This suggests that one needs to control for blinking when comparing detection times between conditions that could elicit phasic changes in blinking.


Can the prostate risk calculator based on Western population be applied to Asian population?
We developed a korean prostate cancer risk calculator (KPCRC) for predicting the probability of a positive initial prostate biopsy using clinical and laboratory data from a Korean male population (http://pcrc.korea.ac.kr). We compared its performance to prostate-specific antigen (PSA) testing and the Prostate Risk Calculator 3 (PRC 3) based on data from the Dutch part of European Randomized Study of Screening for Prostate Cancer (ERSPC), which predicts biopsy results for previously unscreened men. Data were collected from 602 Korean men who were previously unscreened and underwent initial ten-core prostate biopsies. Multiple logistic regression analysis was performed to determine the significant predictors. Area under the receiver operating characteristic curve (AUC) and calibration plots of both calculators were evaluated. Prostate cancer (PCa) was detected in 172 (28.6%) men. Independent predictors of a positive biopsy included advanced age, elevated PSA levels, reduced volume of the transition zone, and abnormal digital rectal examination findings. The AUC of the KPCRC was higher than the PRC 3 and PSA alone on internal and external validation. Calibration plots of the KPCRC showed better performance than the other models on internal and external validation. Applying a cut-off of 10% of KPCRC implied that 251 of the 602 men (42%) would not have been biopsied and that 12 of the 172 PCa cases (7%) would not have been diagnosed. The KPCRC improves the performance of the PRC 3 and PSA testing in predicting Korean population's risk of PCa. It implies that Asian populations need their own risk calculators for PCa.


A multimarker model to predict outcome in tamoxifen-treated breast cancer patients.
This study was designed to produce a model to predict outcome in tamoxifen-treated breast cancer patients based on clinicopathologic features and multiple molecular markers. This was a retrospective study of 324 stage I to III female breast cancer patients treated with tamoxifen for whom standard clinicopathologic data and tumor tissue microarrays were available. Nine molecular markers were studied by semiquantitative immunohistochemistry and/or fluorescence in situ hybridization. Cox proportional hazards analysis was used to determine the contributions of each variable to disease-specific and overall survival, and machine learning was used to produce a model to predict patient outcome. On a univariate basis, the following features were significantly associated with worse survival: high pathologic tumor or nodal class, histologic grade, epidermal growth factor receptor, ERBB2, MYC, or TP53; absent estrogen receptor (ER) or progesterone receptor; and low BCL2. CCND1 and CDKN1B did not reach statistical significance. On a multivariate basis, nodal class, ER, and MYC were statistically significant as independent factors for survival. However, the benefit of ER-positive status was moderated by BCL2, ERBB2, and progesterone receptor. BCL2 and TP53 also interacted as an independent risk factor. A kernel partial least squares polynomial model was developed with an area under the receiver operating characteristic curve of 0.90. Our data show the predictive value of BCL2, ERBB2, MYC, and TP53 in addition to the standard hormone receptors and clinicopathologic features, and they show the importance of conditional interpretation of certain molecular markers. Our multimarker predictive model performed significantly better than standard guidelines.


Telehealth Interventions to Improve Obstetric and Gynecologic Health Outcomes: A Systematic Review.
To systematically review the effectiveness of telehealth interventions for improving obstetric and gynecologic health outcomes. We conducted a comprehensive search for primary literature in ClinicalTrials.gov, Cochrane Library, Cochrane Collaboration Registry of Controlled Trials, EMBASE, PubMed, and MEDLINE. Qualifying primary studies had a comparison group, were conducted in countries ranked very high on the United Nations Human Development Index, published in English, and evaluated obstetric and gynecologic health outcomes. Cochrane Collaboration's tool and ROBINS-I tool were used for assessing risk of bias. Summary of evidence tables were created using the United States Preventive Services Task Force Summary of Evidence Table for Evidence Reviews. Of the 3,926 published abstracts identified, 47 met criteria for inclusion and included 31,967 participants. Telehealth interventions overall improved obstetric outcomes related to smoking cessation and breastfeeding. Telehealth interventions decreased the need for high-risk obstetric monitoring office visits while maintaining maternal and fetal outcomes. One study found reductions in diagnosed preeclampsia among women with gestational hypertension. Telehealth interventions were effective for continuation of oral and injectable contraception; one text-based study found increased oral contraception rates at 6 months. Telehealth provision of medication abortion services had similar clinical outcomes compared with in-person care and improved access to early abortion. Few studies suggested utility for telehealth to improve notification of sexually transmitted infection test results and app-based intervention to improve urinary incontinence symptoms. Telehealth interventions were associated with improvements in obstetric outcomes, perinatal smoking cessation, breastfeeding, early access to medical abortion services, and schedule optimization for high-risk obstetrics. Further well-designed studies are needed to examine these interventions and others to generate evidence that can inform decisions about implementation of newer telehealth technologies into obstetrics and gynecology practice.


Genome-wide annotation and comparative analysis of cuticular protein genes in the noctuid pest Spodoptera litura.
Insect cuticle is considered an adaptable and versatile building material with roles in the construction and function of exoskeleton. Its physical properties are varied, as the biological requirements differ among diverse structures and change during the life cycle of the insect. Although the bulk of cuticle consists basically of cuticular proteins (CPs) associated with chitin, the degree of cuticular sclerotization is an important factor in determining its physical properties. Spodoptera litura, the tobacco cutworm, is an important agricultural pest in Asia. Compared to the domestic silkworm, Bombyx mori, another lepidopteran whose CP genes have been well annotated, S. litura has a shorter life cycle, hides in soil during daytime beginning in the 5th instar and is exposed to soil in the pupal stage without the protection of a cocoon. In order to understand how the CP genes may have been adapted to support the characteristic life style of S. litura, we searched its genome and found 287 putative cuticular proteins that can be classified into 9 CP families (CPR with three groups (RR-1, RR-2, RR-3), CPAP1, CPAP3, CPF, CPFL, CPT, CPG, CPCFC and CPLCA), and a collection of unclassified CPs named CPH. There were also 112 cuticular proteins enriched in Histidine residues with content varying from 6% to 30%, comprising many more His-rich cuticular proteins than B. mori. A phylogenetic analysis between S. litura, M. sexta and B. mori uncovered large expansions of RR-1 and RR-2 CPs, forming large gene clusters in different regions of S. litura chromosome 9. We used RNA-seq analysis to document the expression profiles of CPs in different developmental stages and tissues of S. litura. The comparative genomic analysis of CPs between S. litura and B. mori integrated with the unique behavior and life cycle of the two species offers new insights into their contrasting ecological adaptations.


Relationship between body mass index and renal function deterioration among the Taiwanese chronic kidney disease population.
This study investigated the characteristics of patients with different chronic kidney disease (CKD) stages according to various body mass index (BMI) categories and determined the influence of BMI in renal function deterioration. We conducted a multicenter, longitudinal cohort study based on the Epidemiology and Risk Factors Surveillance of CKD project (2008-2013) and National Health Insurance Research Database (2001-2013). A total of 7357 patients with CKD aged 20-85 years from 14 hospitals were included in the study. A higher male sex, diabetes mellitus (DM) and hypertension were noted among overweight and obese CKD patients, while more cancer prevalence was noted among underweight CKD patients. Charlson comorbidity index was significantly higher and correlated with BMI among late CKD patients. Patients with BMI < 18.5 kg/m2 exhibited non-significantly higher events of eGFR decline events in both early and late CKD stages than other BMI groups. BMI alone is not a determinant of CKD progression among our Taiwanese CKD patients. Obesity should be re-defined and body weight manipulation should be individualized in CKD patients.


A multiancestry study identifies novel genetic associations with CHRNA5 methylation in human brain and risk of nicotine dependence.
Nicotine dependence is influenced by chromosome 15q25.1 single nucleotide polymorphisms (SNPs), including the missense SNP rs16969968 that alters function of the α5 nicotinic acetylcholine receptor (CHRNA5) and noncoding SNPs that regulate CHRNA5 mRNA expression. We tested for cis-methylation quantitative trait loci (cis-meQTLs) using SNP genotypes and DNA methylation levels measured across the IREB2-HYKK-PSMA4-CHRNA5-CHRNA3-CHRNB4 genes on chromosome 15q25.1 in the BrainCloud and Brain QTL cohorts [total N = 175 European-Americans and 65 African-Americans (AAs)]. We identified eight SNPs that were significantly associated with CHRNA5 methylation in prefrontal cortex: P ranging from 6.0 × 10(-10) to 5.6 × 10(-5). These SNP-methylation associations were also significant in frontal cortex, temporal cortex and pons: P ranging from 4.8 × 10(-12) to 3.4 × 10(-3). Of the eight cis-meQTL SNPs, only the intronic CHRNB4 SNP rs11636753 was associated with CHRNA5 methylation independently of the known SNP effects in prefrontal cortex, and it was the most significantly associated SNP with nicotine dependence across five independent cohorts (total N = 7858 European ancestry and 3238 AA participants): P = 6.7 × 10(-4), odds ratio (OR) [95% confidence interval (CI)] = 1.11 (1.05-1.18). The rs11636753 major allele (G) was associated with lower CHRNA5 DNA methylation, lower CHRNA5 mRNA expression and increased nicotine dependence risk. Haplotype analyses showed that rs11636753-G and the functional rs16969968-A alleles together increased risk of nicotine dependence more than each variant alone: P = 3.1 × 10(-12), OR (95% CI) = 1.32 (1.22-1.43). Our findings identify a novel regulatory SNP association with nicotine dependence and connect, for the first time, previously observed differences in CHRNA5 mRNA expression and nicotine dependence risk to underlying DNA methylation differences.


Proteases, extracellular matrix, and cancer: a workshop of the path B study section.
The role of the extracellular matrix (ECM) in the tumor microenvironment is not limited to being a barrier against tumor invasion. The ECM is a reservoir of cell binding proteins and growth factors that affect tumor cell behavior. It is also substantially modified by proteases produced by tumor cells or stroma cells. As a result of the activity of these proteases, cell-cell and cell-ECM interactions are altered, new biologically active ECM molecules are generated, and the bioavailability and activity of many growth factors, growth factor receptors, and cytokines are modified. ECM-degrading proteases also play a critical role in angiogenesis, where they can act as positive as well as negative regulators of endothelial cell proliferation and vascular morphogenesis. This review article summarizes some of the most relevant findings made over the recent years that were discussed at a workshop organized by the Path B Study Section of the National Institutes of Health in October 2002.


Recrudescence of onchocerciasis in the Comoé valley in Southwest Burkina Faso.
Onchocerciasis control by vector control was instigated in southwest Burkina Faso in January 1969 by ORSTOM/OCCGE, and continued until operations were taken over by the WHO Onchocerciasis Control Programme (OCP) in February 1975, which itself ceased operations in the area in 1989 when onchocerciasis was judged to have been reduced to insignificant levels. Initially (1969-1975) vector immigration maintained unacceptably high levels of transmission, but OCP was much larger than the preceding campaign and in 1975 the Annual Transmission Potential (ATP) dropped below 100 at all sites in the Comoé river valley except Folonzo, which continued to be subject to reinvasion, along with the whole of the Léraba river valley. However, after the southern extension of the OCP in 1979, ATPs dropped below 100 everywhere in the Comoé basin (including the Léraba valley), and further dropped to insignificant levels after the western extension of the OCP in 1985. Thus transmission dropped more quickly in the Comoé river valley than the Léraba river valley (which had been subject to vector reinvasion), and this was also reflected in prevalence of microfilaraemia in the human population. After 1986 prevalence was less than 5% in all villages in the Comoé river valley (except for two, which subsequently dropped to 0% and 3.7% by 1999). However, in 2001 (12 years after the cessation of vector control) the prevalence in one village in the Comoé river valley had increased to 39.6%, and two more had increased above 5% by 2007. New epidemiological surveys in 2011 and 2012 showed that in 13 out of 30 villages in the Comoé river valley prevalence of microfilaraemia was above 5%, although this was not observed in the Léraba river valley where prevalence remained low. This is the first documented case of recrudescence of onchocerciasis in the old OCP area, and the reasons are not clear. It is possible that there has been immigration of parasites with humans or vectors from areas where there has been a shorter period of control, or that control has been less effective. It is possible that in spite of very low levels of transmission the local parasite population was never reduced to a level below the transmission breakpoint, or that there has been a local recrudescence due to stochastic population effects. In any case it is clear that the distribution of ivermectin against lymphatic filariasis in the area since 2004 has failed to prevent the recrudescence of onchocerciasis, and the Burkina Faso Programme National de Lutte contre l'Onchocercose (PNLO - Ministere de la Santé) has instigated a programme of Community Directed Treatment with Ivermectin specifically aimed at onchocerciasis in accordance with the strategy developed by APOC and recommended to governments by OCP when it was dissolved in 2002.


Localizing acute stroke-related EEG changes: assessing the effects of spatial undersampling.
Because of its sensitivity to metabolic and ionic disturbances related to ischemia, the EEG can be a potentially useful tool for acute stroke detection and for monitoring affected tissue. However, the clinical use of the EEG in detecting stroke is determined in part by how accurately the spatial information is characterized. The purpose of the current study was to determine the effects of spatial undersampling on the distribution and interpretation of the stroke-related topographic EEG. Using a 128-channel sensor montage, EEG was recorded from six stroke patients acutely (between 8 and 36 hours) after symptom onset. The EEG was submitted to a spectral analysis and was compared with patient symptoms and MRI and computed tomographic findings. To determine loss of spatial and clinical information resulting from spatial undersampling, the average-referenced data from the original 128-channel recording montage were subsampled into 64-, 32-, and 19-channel arrays. Furthermore, the analytical findings were compared with a board-certified electroencephalographer's review of the raw EEG using a conventional clinical montage. As predicted, the results showed that accurate description of stroke-related topographic EEG changes is dependent on adequate spatial sampling density. Accurate description of the spatial distribution of the stroke-related EEG was achieved only with the 64- and 128-channel EEG. As the recording density decreases to 32 channels, the distribution of the scalp EEG spectra is distorted, potentially resulting in mislocalization of the affected region. Results of the clinical review by an expert electroencephalographer corroborated the quantitative analyses, and the results also demonstrated the shortcomings of the conventional 10-20 recording density for capturing focal EEG abnormalities in several cases. The EEG provides useful information about the localization of acute cerebral ischemia, but recording densities of 64 channels or higher are required for accurate spatial characterization of focal stroke-related EEG changes.


Microbiological and geochemical heterogeneity in an in situ uranium bioremediation field site.
The geochemistry and microbiology of a uranium-contaminated subsurface environment that had undergone two seasons of acetate addition to stimulate microbial U(VI) reduction was examined. There were distinct horizontal and vertical geochemical gradients that could be attributed in large part to the manner in which acetate was distributed in the aquifer, with more reduction of Fe(III) and sulfate occurring at greater depths and closer to the point of acetate injection. Clone libraries of 16S rRNA genes derived from sediments and groundwater indicated an enrichment of sulfate-reducing bacteria in the order Desulfobacterales in sediment and groundwater samples. These samples were collected nearest the injection gallery where microbially reducible Fe(III) oxides were highly depleted, groundwater sulfate concentrations were low, and increases in acid volatile sulfide were observed in the sediment. Further down-gradient, metal-reducing conditions were present as indicated by intermediate Fe(II)/Fe(total) ratios, lower acid volatile sulfide values, and increased abundance of 16S rRNA gene sequences belonging to the dissimilatory Fe(III)- and U(VI)-reducing family Geobacteraceae. Maximal Fe(III) and U(VI) reduction correlated with maximal recovery of Geobacteraceae 16S rRNA gene sequences in both groundwater and sediment; however, the sites at which these maxima occurred were spatially separated within the aquifer. The substantial microbial and geochemical heterogeneity at this site demonstrates that attempts should be made to deliver acetate in a more uniform manner and that closely spaced sampling intervals, horizontally and vertically, in both sediment and groundwater are necessary in order to obtain a more in-depth understanding of microbial processes and the relative contribution of attached and planktonic populations to in situ uranium bioremediation.


Coping as a mediator in the relationships of spiritual well-being to mental health in black women with type 2 diabetes.
Examine coping as a mediator in the relationships of spiritual well-being to mental health in Black women with type 2 diabetes (T2DM). Using a cross-sectional design, data were collected from a convenience sample of 45 Black women with T2DM. Measures of coping strategies, spiritual well-being (religious and existential well-being), and mental health, as measured by diabetes-specific distress (DSED), were collected. Bivariate findings informed mediational, trivariate model development. Religious well-being was significantly related to cognitive reframing (CR) coping strategies (p = 0.026) but not DSED (p = 0.751). Existential well-being was significantly related to CR (beta = 0.575,p < 0.001), direct assistance (DA) coping (beta = 0.368, p = 0.006) and DSED (beta = -0.338, p = 0.023). Although CR (beta = -0.305, p = 0.021) and DA (beta = -0.262, p = 0.041) had significant bivariate associations with DSED, the relationships were not significant when existential well-being was controlled. However, the relationship of existential well-being to DSED was mediated by specific CR and DA strategies that were associated with DSED to varying degrees -"I came up with a couple different solutions to the problem" (beta = -0.301, p = 0.049); "I came out of the experience better than I went in" (beta = -0.308, p = 0.061); and "I talked to someone who could do something concrete about the problem" (beta = -0.272, p = 0.078). Findings indicate that diabetes care address spiritual well-being, both its religious and existential components, in Black women with T2DM.


Chromatin remodeling in cardiovascular development and physiology.
Chromatin regulation provides an important means for controlling cardiac gene expression under different physiological and pathological conditions. Processes that direct the development of normal embryonic hearts and pathology of stressed adult hearts may share general mechanisms that govern cardiac gene expression by chromatin-regulating factors. These common mechanisms may provide a framework for us to investigate the interactions among diverse chromatin remodelers/modifiers and various transcription factors in the fine regulation of gene expression, essential for all aspects of cardiovascular biology. Aberrant cardiac gene expression, triggered by a variety of pathological insults, can cause heart diseases in both animals and humans. The severity of cardiomyopathy and heart failure correlates strongly with abnormal cardiac gene expression. Therefore, controlling cardiac gene expression presents a promising approach to the treatment of human cardiomyopathy. This review focuses on the roles of ATP-dependent chromatin-remodeling factors and chromatin-modifying enzymes in the control of gene expression during cardiovascular development and disease.


The immunogenic SigA enterotoxin of Shigella flexneri 2a binds to HEp-2 cells and induces fodrin redistribution in intoxicated epithelial cells.
We have previously shown that the enterotoxin SigA which resides on the she pathogenicity island (PAI) of S. flexneri 2a is an autonomously secreted serine protease capable of degrading casein. We have also demonstrated that SigA is cytopathic for HEp-2 cells and plays a role in the intestinal fluid accumulation associated with S. flexneri infections. In this work we show that SigA binds specifically to HEp-2 cells and degrades recombinant human alphaII spectrin (alpha-fodrin) in vitro, suggesting that the cytotoxic and enterotoxic effects mediated by SigA are likely associated with the degradation of epithelial fodrin. Consistent with our data, this study also demonstrates that SigA cleaves intracellular fodrin in situ, causing its redistribution within cells. These results strongly implicate SigA in altering the cytoskeleton during the pathogenesis of shigellosis. On the basis of these findings, cleavage of fodrin is a novel mechanism of cellular intoxication for a Shigella toxin. Furthermore, information regarding immunogenicity to SigA in infected patients is lacking. We studied the immune response of SigA from day 28 post-challenge serum of one volunteer from S. flexneri 2a challenge studies. Our results demonstrate that SigA is immunogenic following infection with S. flexneri 2a. This work shows that SigA binds to epithelial HEp-2 cells as well as being able to induce fodrin degradation in vitro and in situ, further extending its documented role in the pathogenesis of Shigella infections.


Primary care of people with spinal cord injury: scoping review.
To perform a scoping review of the empirical evidence between 1980 and 2009 regarding primary care for adults with spinal cord injury (SCI). Peer-reviewed journals were searched from 1980 to 2009 using CINAHL, PubMed-MEDLINE, EMBASE, PsycINFO, Social Sciences Abstracts, and Social Work Abstracts. The key word-driven electronic search identified 42 articles on primary care and SCI. Inclusion criteria narrowed the set to 21 articles that were published in English, that had a sample size of greater than 3, and that offered empirical analysis. Approximately 90% of people with SCI identify family physicians as their regular doctors; 63% have SCI specialists. People with long-term SCI develop complex rubrics for navigating their personal health care systems. There is conflicting evidence about the effectiveness of outreach programs for maintaining health and preventing complications following SCI. Regular follow-up by specialized teams and annual comprehensive health examination are supported by the evidence. The research shows a high level of consistency in identifying the most common issues raised by people with SCI in primary care, most of which are related to disability-specifically, secondary complications such as bowel or bladder dysfunction and pain. There is also good evidence that many general health issues require attention in this population, such as bone density problems, depression, and sexual and reproductive health issues. There is level 4 and 5 evidence for unmet health needs among individuals living with SCI in the community. Despite patients with SCI being high users of primary care and health services in general, the evidence suggests that the information needs of these patients in particular are poorly met. A robust system of primary care is the best assurance of good health outcomes and reasonable health service use for people with SCI, including annual comprehensive examination, appropriate specialist use, and attention to accessibility and unmet needs.


Cerebral oxygenation changes in response to motor stimulation.
We studied cerebral hemodynamic response to a sequential motor task in 56 subjects to investigate the time course and distribution of blood oxygenation changes as monitored by near-infrared spectroscopy (NIRS). To address whether response is modulated by different performance velocities, a group of subjects (n = 12) was examined while performing the motor task at 1, 2, and 3 Hz. The results demonstrate that 1) the NIRS response reflects localized changes in cerebral hemodynamics, 2) the response, consisting of an increase in oxygenated hemoglobin concentration [oxy-Hb] and a decrease in deoxygenated hemoglobin concentration ([deoxy-Hb]), is lateralized and increases in amplitude with higher performance rates, and 3) changes in [oxy-Hb] and [deoxy-Hb] differ in time course. Changes in [oxy-Hb] are biphasic, with a fast initial increase and a pronounced poststimulus undershoot. The stimulus-associated decrease in [deoxy-Hb] is monophasic, and response latency is greater. We conclude that NIRS is able to detect even small changes in cerebral hemodynamic response to functional stimulation.


Public health response to two incidents of confirmed MERS-CoV cases travelling on flights through London Heathrow Airport in 2014 – lessons learnt.
In May 2014, Public Health England was alerted to two separate laboratory-confirmed cases of Middle East respiratory syndrome coronavirus (MERS-CoV) infection who transited through London Heathrow Airport while symptomatic on flights from Saudi Arabia to the United States of America. We present the rationale for the public health response to both incidents, and report results of contact tracing. Following a risk assessment, passengers seated two seats around the cases were prioritised for contact tracing and a proactive media approach was used to alert all passengers on the planes of their possible exposure in both incidents. In total, 64 United Kingdom (UK) residents were successfully contacted, 14 of whom were sat in the priority area two seats all around the case(s). Five passengers reported respiratory symptoms within 14 days of the flight, but all tested were negative for MERS-CoV. Details of non-UK residents were passed on to relevant World Health Organization International Health Regulation focal points for follow-up, and no further cases were reported back. Different approaches were used to manage contact tracing for each flight due to variations in the quality and timeliness of the passenger contact information provided by the airlines involved. No evidence of symptomatic onward transmission was found.


In vivo comparison of radiation exposure of dual-energy CT versus low-dose CT versus standard CT for imaging urinary calculi.
Dual-energy computed tomography (DECT) is an emerging imaging modality with the unique capability of determining urinary stone composition. This study compares radiation exposure of DECT, standard single-energy CT (SECT), and low-dose renal stone protocol single-energy CT (LDSECT) for the evaluation of nephrolithiasis in a single in vivo patient cohort. Following institutional review board (IRB) approval, we retrospectively reviewed 200 consecutive DECT examinations performed on patients with suspected urolithiasis over a 6-month period. Of these, 35 patients had undergone examination with our LDSECT protocol, and 30 patients had undergone examination of the abdomen and pelvis with our SECT imaging protocol within 2 years of the DECT examination. The CT dose index volume (CTDIvol) was used to compare radiation exposure between scans. Image quality was objectively evaluated by comparing image noise. Statistical evaluation was performed using a Student's t-test. DECT performed at 80/140 kVp and 100/140 kVp did not produce a significant difference in radiation exposure compared with LDSECT (p=0.09 and 0.18, respectively). DECT performed at 80/140 kVp and 100/140 kVp produced an average 40% and 31%, respectively, reduction in radiation exposure compared with SECT (p<0.001). For patients imaged with the 100/140 kVp protocol, average values for images noise were higher in the LDSECT images compared with DECT images (p<0.001) and there was no significant difference in image noise between DECT and SECT images in the same patient (p=0.88). Patients imaged with the 80/140 kVp protocol had equivocal image noise compared with LDSECT images (p=0.44), however, DECT images had greater noise compared with SECT images in the same patient (p<0.001). Of the 75 patients included in the study, stone material was available for 16; DECT analysis correctly predicted stone composition in 15/16 patients (93%). DECT provides knowledge of stone composition in addition to the anatomic information provided by LDSECT/SECT without increasing patient radiation exposure and with minimal impact on image noise.


When the fourth water and digital revolution encountered COVID-19.
The ongoing COVID-19 pandemic is, undeniably, a substantial shock to our civilization which has revealed the value of public services that relate to public health. Ensuring a safe and reliable water supply and maintaining water sanitation has become ever more critical during the pandemic. For this reason, researchers and practitioners have promptly investigated the impact associated with the spread of SARS-CoV-2 on water treatment processes, focusing specifically on water disinfection. However, the COVID-19 pandemic impacts multiple aspects of the urban water sector besides those related to the engineering processes, including sanitary, economic, and social consequences which can have significant effects in the near future. Furthermore, this outbreak appears at a time when the water sector was already experiencing a fourth revolution, transitioning toward the digitalisation of the sector, which redefines the Water-Human-Data Nexus. In this contribution, a product of collaboration between academics and practitioners from water utilities, we delve into the multiple impacts that the pandemic is currently causing and their possible consequences in the future. We show how the digitalisation of the water sector can provide useful approaches and tools to help address the impact of the pandemic. We expect this discussion to contribute not only to current challenges, but also to the conceptualization of new projects and the broader task of ameliorating climate change.


Patient-centered care and its effect on outcomes in the treatment of asthma.
Patient-centered care may be pivotal in improving health outcomes for patients with asthma. In addition to increased attention in both research and clinical forums, recent legislation also highlights the importance of patient-centered outcomes research in the Patient Protection and Affordable Care Act. However, whether patient-centered care has been shown to improve outcomes for this population is unclear. To answer this question, we performed a systematic review of the literature that aimed to define current patient-focused management issues, characterize important patient-defined outcomes in asthma control, and identify current and emerging treatments related to patient outcomes and perspectives. We used a parallel search strategy via Medline(®), Cochrane Central Register of Controlled Trials, CINAHL(®) (Cumulative Index to Nursing and Allied Health Literature), and PsycINFO(®), complemented with a reference review of key articles that resulted in a total of 133 articles; 58 were interventions that evaluated the effect on patient-centered outcomes, and 75 were descriptive studies. The majority of intervention studies demonstrated improved patient outcomes (44; "positive" results); none showed true harm (0; "negative"); and the remainder were equivocal (14; "neutral"). Key themes emerged relating to patients' desires for asthma knowledge, preferences for tailored management plans, and simplification of treatment regimens. We also found discordance between physicians and patients regarding patients' needs, beliefs, and expectations about asthma. Although some studies show promise regarding the benefits of patient-focused care, these methods require additional study on feasibility and strategies for implementation in real world settings. Further, it is imperative that future studies must be, themselves, patient-centered (eg, pragmatic comparative effectiveness studies) and applicable to a variety of patient populations and settings. Despite the need for further research, enough evidence exists that supports incorporating a patient-centered approach to asthma management, in order to achieve improved outcomes and patient health.


Binocular Vision, Visual Function, and Pupil Dynamics in People Living With Dementia and Their Relation to the Rate of Cognitive Decline and Structural Changes Within the Brain: Protocol for an Observational Study.
Visual impairment is a common comorbidity in people living with dementia. Addressing sources of visual difficulties can have a significant impact on the quality of life for people living with dementia and their caregivers. Depth perception problems are purportedly common in dementia and also contribute to falls, visuomotor task difficulties, and poorer psychosocial well-being. However, depth perception and binocular vision are rarely assessed in dementia research. Sleep fragmentation is also common for people living with dementia, and binocular cooperation for depth perception can be affected by fatigue. Pupillary responses under cognitive load also have the potential to be a risk marker for cognitive decline in people living with dementia and can be combined with the above measures for a comprehensive evaluation of clinical visual changes in people living with dementia and their relation to changes in cognitive status, sleep quality, and cortical structure or function. This study aims to characterize the nature of clinical visual changes and altered task-evoked pupillary responses that may occur in people living with dementia and evaluate whether these responses relate to changes in cognitive status (standardized Mini Mental State Examination [MMSE] score), Pittsburgh sleep quality index, and cortical structure or function. This proposed exploratory observational study will enroll ≤210 people with recently diagnosed dementia (within the last 24 months). The following parameters will be assessed on 3 occasions, 4 months apart (plus or minus 2 weeks): visual function (visual acuity and contrast sensitivity), binocular function (motor fusion and stereopsis), task-evoked pupillary responses (minimum and maximum pupil size, time to maximum dilation, and dilation velocity), cognitive status (MMSE score), and sleep quality (Pittsburgh Sleep Quality Index). A subset of patients (n=30) with Alzheimer disease will undergo structural and functional magnetic resonance imaging at first and third visits, completing a 10-day consensus sleep diary to monitor sleep quality, verified by sleep actimetry. This research was funded in February 2018 and received National Health Service Research Ethics Committee approval in September 2018. The data collection period was from October 1, 2018, to November 30, 2019. A total of 24 participants were recruited for the study. The data analysis is complete, with results expected to be published before the end of 2020. Findings will demonstrate how often people with dementia experience binocular vision problems. If frequent, diagnosing and treating them could improve quality of life by reducing the risk of falls and fine visuomotor task impairment and by relieving psychosocial anxiety. This research will also demonstrate whether changes in depth perception, pupillary responses, and quality of vision relate to changes in memory or sleep quality and brain structure or function. If related, these quick and noninvasive eye tests help monitor dementia. This would help justify whether binocular vision and pupillary response testing should be included in dementia-friendly eye-testing guidelines. RR1-10.2196/16089.


A Taenia crassiceps factor induces apoptosis of spleen CD4+T cells and TFG-β and Foxp3 gene expression in mice.
This study was undertaken to determine whether a parasite substance produces structural pathology in the mouse spleen. A low-molecular-weight Taenia crassiceps metacestode factor (MF) isolated from the peritoneal fluid of female mice infected with T. crassiceps metacestodes induced pathological and immunological changes in mouse spleen cells in vivo. Electron microscopy and confocal microscopy revealed severe changes in the spleen histoarchitecture of T. crassiceps-infected and MF-treated mice. Apoptotic degenerated spleen cells were observed in the white and red pulps and were more conspicuous in the white pulp of the spleen from the T. crassiceps-infected mice than in that of the MF-treated mice. Flow cytometry analysis revealed that the numbers of spleen CD4+T cells were significantly lower in both experimental groups than in control mice. The ex vivo expression of transforming growth factor (TGF)-β and factor Foxp3 were significantly higher in splenocytes of the experimental mice than the basal expression observed in the control cells. These findings may have potential applications for a better understanding of the host-parasite relationship in human neurocysticercosis.


IL-18 induction of osteopontin mediates cardiac fibrosis and diastolic dysfunction in mice.
Osteopontin (OPN), a key component of the extracellular matrix, is associated with the fibrotic process during tissue remodeling. OPN and the cytokine interleukin (IL)-18 have been shown to be overexpressed in an array of human cardiac pathologies. In the present study, we determined the role of IL-18 in the regulation of cardiac OPN expression and the subsequent interstitial fibrosis and diastolic dysfunction. We demonstrated parallel increases in IL-18, OPN expression, and interstitial fibrosis in murine models of left ventricular pressure and volume overload. Exogenous recombinant (r)IL-18 administered for 2 wk increased cardiac OPN expression, interstitial fibrosis, and diastolic dysfunction. Stimulation of the T helper (Th)1 lymphocyte phenotype with a selective toll-like receptor (TLR)9 agonist induced cardiac IL-18 and OPN expression, which was associated with increased cardiac fibrillar collagen concentrations and interstitial fibrosis resulting in diastolic dysfunction. rIL-18 induced OPN expression and protein levels in primary of cardiac fibroblast cultures. Conditioned media from TLR9-stimulated T lymphocyte cultures induced IL-18 and OPN expression in cardiac fibroblasts, while blockade of the IL-18 receptor with a neutralizing antibody abolished the increase in OPN expression. Furthermore, a mutation in the transcriptional factor interferon regulatory factor (IRF)1 or IRF1 small interfering RNA (siRNA) resulted in the decreased expression of IL-18 and OPN in cardiac fibroblasts. With pressure overload, IRF1-mutant mice showed downregulation of IL-18 and OPN expression in cardiac tissue, reduced cardiac fibrotic development, and increased left ventricular function compared with wild type. These results provide direct evidence that the induction of IL-18 regulates OPN-mediated cardiac fibrosis and diastolic dysfunction.


The effect of alcohol on cervical and ocular vestibular evoked myogenic potentials in healthy volunteers.
We investigated the effect of alcohol on the cervical and ocular vestibular evoked myogenic potentials (cVEMPs and oVEMPs). As alcohol produces gaze-evoked nystagmus (GEN), we also tested the effect of nystagmus independent of alcohol by recording oVEMPs during optokinetic stimulation (OKS). The effect of alcohol was tested in 14 subjects over multiple rounds of alcohol consumption up to a maximum breath alcohol concentration (BrAC) of 1.5‰ (mean 0.97‰). The effect of OKS was tested in 11 subjects at 5, 10 and 15deg/sec. oVEMP amplitude decreased from baseline to the highest BrAC level by 27% (range 5-50%, P<0.001), but there was no significant effect on oVEMP latency or cVEMP amplitude or latency. There was a significant negative effect of OKS on oVEMP amplitude (16%, P=0.006). We found a selective effect of alcohol on oVEMP amplitude, but no effect on the cVEMP. Vertical nystagmus elicited by OKS reduced oVEMP amplitude. Alcohol selectively affects oVEMP amplitude. Despite the effects of alcohol and nystagmus, both reflexes were reliably recorded in all subjects and conditions. An absent response in a patient affected by alcohol or nystagmus indicates a vestibular deficit.


Effects of Bay o 2752, a hypocholesterolemic agent, on intestinal taurocholate absorption and cholesterol esterification.
Bay o 2752 [N,N'-(1, 11-undecandiyl)bis(2,3-dihydro-2-methyl-1H-indole-1-carboxamide)] has been demonstrated in rats to inhibit intestinal cholesterol absorption. Studies were carried out in male Wistar rats to determine if the mechanism is inhibition of intestinal bile acid absorption or cholesterol esterification. Bay o 2752 did not alter intestinal bile acid absorption as measured by in vitro uptake of [14C]taurocholic acid into ileal everted sacs (0.01 and 1.0 mg/ml of Bay o 2752) or the biliary excretion of radioactivity after in vivo ileal perfusion of the bile acid and drug (1.0 mg/ml at 1.0 ml/min for 1 hr). Cholesterol esterification was determined by measurement of in vitro activity of acyl coenzyme A:cholesterol acyltransferase from hepatic microsomes and cholesterol ester hydrolase from pancreatic supernatant, and the in vivo lymphatic output of cholesteryl ester after intraduodenal cholesteryl infusion. Addition of Bay o 2752 (0.01-10 micrograms/ml) to hepatic microsomes produced a concentration-dependent decrease in acyl coenzyme A:cholesterol acyltransferase activity with an IC50 of 0.95 micrograms/ml. Cholesterol ester hydrolase activity was unaffected by the drug (1.0-100 micrograms/ml). Intraduodenal infusion of Bay o 2752 (10 mg/ml at 0.9 ml/hr for 8 hr) reduced markedly the flux of cholesterol from the intestinal lumen into the mesenteric lymph, especially the lymphatic output of the esterified form of both radioisotopically labeled and total cholesterol. These data suggest that Bay o 2752-induced reduction in intestinal cholesterol absorption results from its potent inhibitory effect on acyl coenzyme A:cholesterol acyltransferase activity.


Impaired ketogenesis and increased acetyl-CoA oxidation promote hyperglycemia in human fatty liver.
Non-alcoholic fatty liver disease (NAFLD) is a highly prevalent, and potentially morbid, disease that affects one-third of the U.S. population. Normal liver safely accommodates lipid excess during fasting or carbohydrate restriction by increasing their oxidation to acetyl-CoA and ketones, yet lipid excess during NAFLD leads to hyperglycemia and, in some, steatohepatitis. To examine potential mechanisms, flux through pathways of hepatic oxidative metabolism and gluconeogenesis were studied using five simultaneous stable isotope tracers in ketotic (24-hour fast) individuals with a wide range of hepatic triglyceride contents (0-52%). Ketogenesis was progressively impaired as hepatic steatosis and glycemia worsened. Conversely, the alternative pathway for acetyl-CoA metabolism, oxidation in the tricarboxylic (TCA) cycle, was upregulated in NAFLD as ketone production diminished and positively correlated with rates of gluconeogenesis and plasma glucose concentrations. Increased respiration and energy generation that occurred in liver when β-oxidation and TCA cycle activity were coupled may explain these findings, inasmuch as oxygen consumption was higher during fatty liver and highly correlated with gluconeogenesis. These findings demonstrate that increased glucose production and hyperglycemia in NAFLD is not a consequence of acetyl-CoA production per se, but how acetyl-CoA is further metabolized in liver.


Hypoglycemic Activity through a Novel Combination of Fruiting Body and Mycelia of Cordyceps militaris in High-Fat Diet-Induced Type 2 Diabetes Mellitus Mice.
Diabetes mellitus (DM) is currently ranked among leading causes of death worldwide in which type 2 DM is reaching an epidemic proportion. Hypoglycemic medications for type 2 DM have either proven inadequate or posed adverse effects; therefore, the Chinese herbal products are under investigation as an alternative treatment. In this study, a novel combination of fruiting body and mycelia powder of herbal Cordyceps militaris number 1 (CmNo1) was administered to evaluate their potential hypoglycemic effects in high-fat diet- (HFD-) induced type 2 DM in C57BL/6J mice. Body weight, fasting blood glucose (FBG), oral glucose tolerance test (OGTT), and blood biochemistry indexes were measured. Results indicated that CmNo1 lowered the blood glucose level by increasing insulin sensitivity, while no change in body weight was observed. Increased protein expression of IRS-1, pIRS-1, AKT, pAKT, and GLUT-4 in skeletal muscle and adipose tissue was found indicating restoration of insulin signaling. Additionally, PPAR-γ expression in adipose tissue restored the triglyceride and cholesterol levels. Finally, our results suggest that CmNo1 possesses strong hypoglycemic, anticholesterolemic, and antihypertriglyceridemic actions and is more economical alternate for DM treatment.


Taxol-induced microtubule asters in mitotic extracts of Xenopus eggs: requirement for phosphorylated factors and cytoplasmic dynein.
Taxol, a microtubule stabilizing drug, induces the formation of numerous microtubule asters in the cytoplasm of mitotic cells (De Brabander, M., G. Geuens, R. Nuydens, R. Willebrords, J. DeMey. 1981. Proc. Natl. Acad. Sci. USA. 78:5608-5612). The center of these asters share with spindle poles some characteristics such as the presence of centrosomal material and calmodulin. We have recently reproduced the assembly of taxol asters in a cell-free system (Buendia, B., C. Antony, F. Verde, M. Bornens, and E. Karsenti. 1990. J. Cell Sci. 97:259-271) using extracts of Xenopus eggs. In this paper, we show that taxol aster assembly requires phosphorylation, and that they do not grow from preformed centers, but rather by a reorganization of microtubules first crosslinked into bundles. This process seems to involve sliding of microtubules along each other and we show that cytoplasmic dynein is required for taxol aster assembly. This result provides a possible functional basis to the recent findings, that dynein is present in the spindle and enriched near spindle poles (Pfarr, C. M., M. Cove, P. M. Grissom, T. S. Hays, M. E. Porter, and J. R. McIntosh. 1990. Nature (Lond.). 345:263-265; Steuer, E. R., L. Wordeman, T. A. Schroer, and M. P. Sheetz. 1990. Nature (Lond.). 345:266-268).


Long-chain conversion of [13C]linoleic acid and alpha-linolenic acid in response to marked changes in their dietary intake in men.
We studied the long-chain conversion of [U-13C]alpha-linolenic acid (ALA) and linoleic acid (LA) and responses of erythrocyte phospholipid composition to variation in the dietary ratios of 18:3n-3 (ALA) and 18:2n-6 (LA) for 12 weeks in 38 moderately hyperlipidemic men. Diets were enriched with either flaxseed oil (FXO; 17 g/day ALA, n=21) or sunflower oil (SO; 17 g/day LA, n=17). The FXO diet induced increases in phospholipid ALA (>3-fold), 20:5n-3 [eicosapentaenoic acid (EPA), >2-fold], and 22:5n-3 [docosapentaenoic acid (DPA), 50%] but no change in 22:6n-3 [docosahexanoic acid (DHA)], LA, or 20:4n-6 [arachidonic acid (AA)]. The increases in EPA and DPA but not DHA were similar to those in subjects given the SO diet enriched with 3 g of EPA plus DHA from fish oil (n=19). The SO diet induced a small increase in LA but no change in AA. Long-chain conversion of [U-13C]ALA and [U-13C]LA, calculated from peak plasma 13C concentrations after simple modeling for tracer dilution in subsets from the FXO (n=6) and SO (n=5) diets, was similar but low for the two tracers (i.e., AA, 0.2%; EPA, 0.3%; and DPA, 0.02%) and varied directly with precursor concentrations and inversely with concentrations of fatty acids of the alternative series. [13C]DHA formation was very low (<0.01%) with no dietary influences.


Serotonin and dopamine play complementary roles in gambling to recover losses.
Continued gambling to recover losses--'loss chasing'--is a prominent feature of social and pathological gambling. However, little is known about the neuromodulators that influence this behavior. In three separate experiments, we investigated the role of serotonin activity, D(2)/D(3) receptor activity, and beta-adrenoceptor activity on the loss chasing of age and IQ-matched healthy adults randomized to treatment or an appropriate control/placebo. In Experiment 1, participants consumed amino-acid drinks that did or did not contain the serotonin precursor, tryptophan. In Experiment 2, participants received a single 176 μg dose of the D(2)/D(3) receptor agonist, pramipexole, or placebo. In Experiment 3, participants received a single 80 mg dose of the beta-adrenoceptor blocker, propranolol, or placebo. Following treatment, participants completed a computerized loss-chasing game. Mood and heart rate were measured at baseline and following treatment. Tryptophan depletion significantly reduced the number of decisions made to chase losses, and the number of consecutive decisions to chase, in the absence of marked changes in mood. By contrast, pramipexole significantly increased the value of losses chased and diminished the value of losses surrendered. Propranolol markedly reduced heart rate, but produced no significant changes in loss-chasing behavior. Loss chasing can be thought of as an aversively motivated escape behavior controlled, in part, by the marginal value of continued gambling relative to the value of already accumulated losses. Serotonin and dopamine appear to play dissociable roles in the tendency of individuals to gamble to recover, or to seek to 'escape' from, previous losses. Serotonergic activity seems to promote the availability of loss chasing as a behavioral option, whereas D(2)/D(3) receptor activity produces complex changes in the value of losses judged worth chasing. Sympathetic arousal, at least as mediated by beta-adrenoceptors, does not play a major role in laboratory-based loss-chasing choices.


Neonatal imitation in context: Sensorimotor development in the perinatal period.
More than 35 years ago, Meltzoff and Moore (1977) published their famous article, "Imitation of facial and manual gestures by human neonates." Their central conclusion, that neonates can imitate, was and continues to be controversial. Here, we focus on an often-neglected aspect of this debate, namely, neonatal spontaneous behaviors themselves. We present a case study of a paradigmatic orofacial "gesture," namely tongue protrusion and retraction (TP/R). Against the background of new research on mammalian aerodigestive development, we ask: How does the human aerodigestive system develop, and what role does TP/R play in the neonate's emerging system of aerodigestion? We show that mammalian aerodigestion develops in two phases: (1) from the onset of isolated orofacial movements in utero to the postnatal mastery of suckling at 4 months after birth; and (2) thereafter, from preparation to the mastery of mastication and deglutition of solid foods. Like other orofacial stereotypies, TP/R emerges in the first phase and vanishes prior to the second. Based upon recent advances in activity-driven early neural development, we suggest a sequence of three developmental events in which TP/R might participate: the acquisition of tongue control, the integration of the central pattern generator (CPG) for TP/R with other aerodigestive CPGs, and the formation of connections within the cortical maps of S1 and M1. If correct, orofacial stereotypies are crucial to the maturation of aerodigestion in the neonatal period but also unlikely to co-occur with imitative behavior.


Interaction of St John's wort with conventional drugs: systematic review of clinical trials.
To determine the methodological quality of clinical trials that examined possible interactions of St John's wort with conventional drugs, and to examine the results of these trials. Systematic review. Electronic databases from inception to April 2004, reference lists from published reports, and experts in the field. Eligible studies were prospective clinical trials evaluating the pharmacokinetic effect of St John's wort on the metabolism of conventional drugs. Two reviewers selected studies for inclusion and independently extracted data. 22 pharmacokinetic trials studied an average of 12 (SD 5) participants; 17 trials studied healthy volunteers and five studied patients. Most (17) studies used a "before and after" design; four studies used control groups other than the active group. Three studies randomised the sequence of administration or the participants to study arms or periods; three studies blinded participants or investigators. In 15 trials, investigators independently assayed the herb. Of 19 trials with available plasma data, three found no important interaction (change in area under the curve < 20%) and 17 found a decrease in systemic bioavailability of the conventional drug; in seven studies the 95% confidence interval excluded a decrease of < 20%. Clinicians and patients should beware of possible decreases in the systemic bioavailability of conventional drugs when taken concomitantly with St John's wort.


Direct genetic demonstration of G alpha 13 coupling to the orphan G protein-coupled receptor G2A leading to RhoA-dependent actin rearrangement.
G2A is an orphan G protein-coupled receptor (GPCR), expressed predominantly in T and B cells and homologous to a small group of GPCRs of unknown function expressed in lymphoid tissues. G2A is transcriptionally induced in response to diverse stimuli, and its ectopic expression suppresses transformation of B lymphoid precursors by BCR-ABL. G2A induces morphological transformation of NIH 3T3 fibroblasts. Microinjection of constructs encoding G2A into Swiss 3T3 fibroblasts induces actin reorganization into stress fibers that depends on RhoA, but not CDC42 or RAC. G2A elicits RhoA-dependent transcriptional activation of serum response factor. Direct evaluation of RhoA activity demonstrates elevated levels of RhoA-GTP in G2A-expressing cells. Microinjection of embryonic fibroblasts derived from various G alpha knockout mice establishes a requirement for G alpha 13 but not G alpha 12 or G alpha q/11 in G2A-induced actin rearrangement. In conclusion, G2A represents a family of GPCRs expressed in lymphocytes that may link diverse stimuli to cytoskeletal reorganization and transcriptional activation through a pathway involving G alpha 13 and RhoA.


[Higher brain dysfunction--from the administrative viewpoint].
The "higher brain dysfunction" illustrates various cognitive and behavioral consequences resulted from organic brain damage--attention deficits, memory disorders, executive dysfunction, and disorders of emotion and social communication. These deficits are hard to be visible externally, so patients with such "higher brain problems" are difficult to be appropriately evaluated and treated even in psychiatry service. The patients who have reached to a chronic phase and whose treatments are terminated have been able to live some stable life at home, but exhibit many difficulties in social life, so they remain unemployment. In Japan, the Ministry of Health, Labour and Welfare has started the project for the investigation and the support of patients with higher brain dysfunction since 2001. In this article, first, the diagnostic criterion for "higher brain dysfunction" is presented. Secondly, each cognitive and behavioral disorder is reviewed. Then, the present state and future issues are indicated. We would emphasize that for the patients with such higher brain dysfunctions, the appropriate treatments and services from the injury to the employment should be ensure, and that multi-disciplinary collaboration including medical doctors and paramedical staff--not only occupational therapist (OT), physical therapist (PT), speech therapist (ST), and clinical psychotherapist but also the experts at neuropsychology and at applied behavior analysis is crucial for overcome patients' problems.


Purification of monoclonal antibody against Ebola GP1 protein expressed in Nicotiana benthamiana.
Monoclonal antibodies (mAbs) are one of the fastest growing drug molecules targeting the treatment of diseases ranging from arthritis, immune disorders, and infectious diseases to cancer. Due to its unique application principle, antibodies are commonly produced in large quantities. Plants, such as Nicotiana benthamiana, offer a unique production platform for bio-therapeutics due to their ability to produce large amounts of biomolecules in a relatively quick manner. However, purification of a target protein from plant is an arduous task due to the presence of toxic compounds in ground plant tissue and the large quantities of plant tissues to be processed. Here, a process was developed prior to the chromatographic purification of a mAb against Ebola GP1 protein expressed in N. benthamiana. The process includes a diafiltration step and a charged polyelectrolyte precipitation. The diafiltration step significantly improved the precipitation efficiency, reducing the usage of polyelectrolyte by more than 2000 fold while improving the native plant protein removed from 60% to 80%. The mAb can then be purified to near homogeneity judging from SDS-PAGE by either Protein A affinity chromatography or a tandem of hydrophobic interaction chromatography and a hydrophobic charge induction chromatography. The purified mAbs were shown to retain their binding specificity to irradiated Ebola virus.


Fusarium Canker of Bitternut Hickory Caused by Fusarium solani in the North-Central and Northeastern United States.
Multiple annual cankers were observed on the upper main stems of bitternut hickory (Carya cordiformis) exhibiting top dieback in Indiana, Iowa, Minnesota, New York, Ohio, and Wisconsin during a 2006 to 2008 survey of declining hickory. The top-killed trees had normal-sized, green leaves below and the cankers were oval, sunken, and bounded by heavy callus that seemed to arrest further canker expansion. Fusarium solani was consistently isolated from the margins of inner bark lesions or discolored sapwood of the cankers. When cultured on potato dextrose agar, the isolates grew rapidly with abundant aerial mycelium. On carnation leaf agar, thick-walled macroconidia with 4 to 5 septa were produced in cream, blue-green, or blue sporodochia. Macroconidia were generally cylindrical with a blunt or rounded apical cell and a rounded or foot-shaped basal cell. Microconidia were oval to kidney shaped with 0 to 1 septa and were produced in false heads on elongate monophialides. Chlamydospores were formed singly or in pairs. These morphological characteristics are consistent with descriptions of F. solani (2). The identities of 42 representative isolates were confirmed by sequencing the translation elongation factor (tef) 1-α gene. BLAST analysis of the sequences from each isolate against the GenBank and FUSARIUM-ID database found 98 to 100% similarities to F. solani isolates (GenBank Accession Nos. DQ246841, DQ247025, DQ247282, and DQ247436 and FUSARIUM-ID isolate FD01041). Two haplotypes (BB and BC) were distinguished based on the tef 1-α gene sequences that differed by 10 bp. Pathogenicity tests were conducted with two isolates of each haplotype on asymptomatic C. cordiformis (12 to 21 cm in diameter) in forest stands. In May 2009 in Wabasha County, MN, 0.1-ml spore suspensions (1 × 104 macroconidia/ml) or sterile water was placed in one of three holes (0.6 cm in diameter) drilled to the cambium of 12 trees. The holes were sealed with moist cotton and moldable putty. A duplicate trial, but with BB and BC isolates from Wisconsin, was initiated in Chippewa County, WI in June 2009. The extent of inner bark necrosis was assessed 13 months after inoculation in both sites. Inoculations with F. solani in Minnesota resulted in inner bark lesions with average lengths of 20 and 30 mm for the BB and BC haplotypes, respectively. In Wisconsin, BB and BC haplotypes caused inner bark lesions with average lengths of 34 and 38 mm, respectively. While sunken or open cankers were found for all the BC isolate inoculations, relatively small and callus-bounded cankers were found for BB isolate inoculations. All control wounds were callus-closed with average wound lengths of 12 and 23 mm in Minnesota and Wisconsin, respectively. The same haplotype of F. solani used for inoculation was recovered from each canker as confirmed by analysis of tef 1-α gene sequences. F. solani was not obtained from control wounds. To our knowledge, this is the first report of a canker caused by F. solani on bitternut hickory (1). The same fungus has been previously reported to cause cankers on stems of other hardwood tree genera in the eastern United States and Canada. We hypothesize that numerous main-stem cankers caused by F. solani lead to top dieback of bitternut hickory. References: (1) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006.


Optimal achromatic wave retarders using two birefringent wave plates: reply.
This reply attempts to cast some more light on the achromatic systems composed by wave plates, in particular to the calculus of the overall retardation and the use of the Jones matrix equivalence theorem. An equivalent expression for the overall retardation of the system in terms of the trace is also given.


The immunological component of the cellular inflammatory infiltrate in bronchiectasis.
Immunohistological analysis of bronchial biopsy specimens from nine patients with bronchiectasis and four control subjects was performed with a panel of monoclonal antibodies selected to show lymphocyte and macrophage subsets and signs of cellular activation. The cells taking part in the inflammatory response in the bronchial wall of patients with bronchiectasis were almost exclusively mononuclear cells, most of them T lymphocytes. B lymphocytes were observed in biopsy specimens from only two out of nine patients. CD8+ T cells outnumbered CD4+ cells in all patients in a ratio ranging from 2:1 to 10:1. Most T lymphocytes also strongly expressed CD7 antigen and a proportion of them expressed HLA-DR. Most of the lymphocytic infiltration occurred just beneath the basement membrane of the epithelium, though intraepithelial and submucosal infiltration was also seen. Non-lymphoid mononuclear cells expressing the phenotype of dendritic cells and macrophages were found dispersed throughout the infiltrate, most of them expressing HLA-DR. These observations support the hypothesis that cell mediated immunological reactions contribute to the inflammation associated with bronchiectasis.


Strain features and distributions in pneumococci from children with invasive disease before and after 13-valent conjugate vaccine implementation in the USA.
The effect of second-generation pneumococcal conjugate vaccines on invasive pneumococcal disease (IPD) strain distributions have not yet been well described. We analysed IPD isolates recovered from children aged <5 years through Active Bacterial Core surveillance before (2008-2009; n = 828) and after (2011-2013; n = 600) 13-valent pneumococcal conjugate vaccine (PCV13) implementation. We employed conventional testing, PCR/electrospray ionization mass spectrometry and whole genome sequence (WGS) analysis to identify serotypes, resistance features, genotypes, and pilus types. PCV13, licensed in February 2010, effectively targeted all major 19A and 7F genotypes, and decreased antimicrobial resistance, primarily owing to removal of the 19A/ST320 complex. The strain complex contributing most to the remaining β-lactam resistance during 2011-2013 was 35B/ST558. Significant emergence of non-vaccine clonal complexes was not evident. Because of the removal of vaccine serotype strains, positivity for one or both pilus types (PI-1 and PI-2) decreased in the post-PCV13 years 2011-2013 relative to 2008-2009 (decreases of 32-55% for PI-1, and >95% for PI-2 and combined PI-1 + PI-2). β-Lactam susceptibility phenotypes correlated consistently with transpeptidase region sequence combinations of the three major penicillin-binding proteins (PBPs) determined through WGS analysis. Other major resistance features were predictable by DNA signatures from WGS analysis. Multilocus sequence data combined with PBP combinations identified progeny, serotype donors and recipient strains in serotype switch events. PCV13 decreased the frequency of all PCV13 serotype clones and concurrently decreased the frequency of strain subsets with resistance and/or adherence features conducive to successful carriage. Our results serve as a reference describing key features of current paediatric IPD strains in the USA after PCV13 implementation.


Light Microscopy of Mitochondria at the Nanoscale.
Mitochondria are essential for eukaryotic life. These double-membrane organelles often form highly dynamic tubular networks interacting with many cellular structures. Their highly convoluted contiguous inner membrane compartmentalizes the organelle, which is crucial for mitochondrial function. Since the diameter of the mitochondrial tubules is generally close to the diffraction limit of light microscopy, it is often challenging, if not impossible, to visualize submitochondrial structures or protein distributions using conventional light microscopy. This renders super-resolution microscopy particularly valuable, and attractive, for studying mitochondria. Super-resolution microscopy encompasses a diverse set of approaches that extend resolution, as well as nanoscopy techniques that can even overcome the diffraction limit. In this review, we provide an overview of recent studies using super-resolution microscopy to investigate mitochondria, discuss the strengths and opportunities of the various methods in addressing specific questions in mitochondrial biology, and highlight potential future developments.


Biomimetic composites with enhanced toughening using silk-inspired triblock proteins and aligned nanocellulose reinforcements.
Silk and cellulose are biopolymers that show strong potential as future sustainable materials. They also have complementary properties, suitable for combination in composite materials where cellulose would form the reinforcing component and silk the tough matrix. A major challenge concerns balancing structure and functional properties in the assembly process. We used recombinant proteins with triblock architecture, combining structurally modified spider silk with terminal cellulose affinity modules. Flow alignment of cellulose nanofibrils and triblock protein allowed continuous fiber production. Protein assembly involved phase separation into concentrated coacervates, with subsequent conformational switching from disordered structures into β sheets. This process gave the matrix a tough adhesiveness, forming a new composite material with high strength and stiffness combined with increased toughness. We show that versatile design possibilities in protein engineering enable new fully biological materials and emphasize the key role of controlled assembly at multiple length scales for realization.


Major medical outcomes with spinal augmentation vs conservative therapy.
The symptomatic benefits of spinal augmentation (vertebroplasty or kyphoplasty) for the treatment of osteoporotic vertebral compression fractures are controversial. Recent population-based studies using medical billing claims have reported significant reductions in mortality with spinal augmentation compared with conservative therapy, but in nonrandomized settings such as these, there is the potential for selection bias to influence results. To compare major medical outcomes following treatment of osteoporotic vertebral fractures with spinal augmentation or conservative therapy. Additionally, we evaluate the role of selection bias using preprocedure outcomes and propensity score analysis. Retrospective cohort analysis of Medicare claims for the 2002-2006 period. We compared 30-day and 1-year outcomes in patients with newly diagnosed vertebral fractures treated with spinal augmentation (n = 10,541) or conservative therapy (control group, n = 115,851). Outcomes were compared using traditional multivariate analyses adjusted for patient demographics and comorbid conditions. We also used propensity score matching to select 9017 pairs from the initial groups to compare the same outcomes. Spinal augmentation (vertebroplasty or kyphoplasty) or conservative therapy. Mortality, major complications, and health care utilization. Using traditional covariate adjustments, mortality was significantly lower in the augmented group than among controls (5.2% vs 6.7% at 1 year; hazard ratio, 0.83; 95% CI, 0.75-0.92). However, patients in the augmented group who had not yet undergone augmentation (preprocedure subgroup) had lower rates of medical complications 30 days post fracture than did controls (6.5% vs 9.5%; odds ratio, 0.66; 95% CI, 0.57-0.78), suggesting that the augmented group was less medically ill. After propensity score matching to better account for selection bias, 1-year mortality was not significantly different between the groups. Furthermore, 1-year major medical complications were also similar between the groups, and the augmented group had higher rates of health care utilization, including hospital and intensive care unit admissions and discharges to skilled nursing facilities. After accounting for selection bias, spinal augmentation did not improve mortality or major medical outcomes and was associated with greater health care utilization than conservative therapy. Our results also highlight how analyses of claims-based data that do not adequately account for unrecognized confounding can arrive at misleading conclusions.


Protective effect of the 1742(C/G) polymorphism of human cardiotrophin-1 against left ventricular hypertrophy in essential hypertension.
Experimental and clinical evidence supports a role of cardiotrophin-1 (CT-1) in the development of hypertensive left ventricular hypertrophy (LVH). The goal of this study was to investigate the relationship between human CT-1 genetic background and LVH in essential hypertension. A total of 900 individuals were genotyped for the 1742(C/G) polymorphism of the human CT-1 gene. Serum CT-1 levels were assessed by ELISA in 681 individuals. Left ventricular parameters were determined by two-dimensional echocardiography in 297 individuals. The prevalence of the GG genotype of the 1742(C/G) polymorphism was reduced in essential hypertension (8.4% in normotensive individuals, 4.9% in hypertensive patients, P = 0.046 versus CC/CG individuals) and in LVH (11.5% in nonhypertrophic normotensive individuals, 12.2% in nonhypertrophic hypertensive patients, 2.6% in hypertensive patients with LVH, P = 0.008 versus CC/CG individuals). Apart from this, GG individuals presented lower serum concentration of CT-1 (GG, 147.1 ± 10.5 fmol/ml; CC/CG, 187.1 ± 4.8 fmol/ml; P = 0.036) and left ventricular mass index (GG, 91 ± 6 g/m; CC/CG, 119 ± 3 g/m; P = 0.002). Multivariate analyses showed that the 1742(C/G) polymorphism was a significant determinant of both left ventricular mass index and serum CT-1, after adjusting for confounding factors. Finally, in-vitro studies supported the functionality of the 1742(C/G) polymorphism. Our results indicate that the 1742(C/G) polymorphism of the human CT-1 gene is associated with LVH in hypertension and that the GG genotype may have a protective role. It is suggested that CT-1 is one of the mediators of this association.