Does the interaction between partnership status and average progesterone level predict women's preferences for facial masculinity?
Many studies have attempted to identify biological factors that reliably predict individual differences in women's preferences for masculine male faces. Marcinkowska et al. (2018, Hormones & Behavior) recently reported that women's (N = 102) preferences for facial masculinity were predicted by the interaction between their relationship status (partnered versus unpartnered) and average progesterone level. Because previous findings for between-women differences in masculinity preferences have often not replicated well, we attempted to replicate Marcinkowska et al.'s result in an open data set from another recent study that had not tested this hypothesis (Jones et al., 2018, Psychological Science). In this sample of 316 women, we found that facial masculinity preferences were predicted by the interaction between women's relationship status and average progesterone level, consistent with Marcinkowska et al.'s results (data and analysis code are available at https://osf.io/q9szc). Together, these findings suggest that the combined effects of relationship status and average progesterone level may predict facial masculinity preferences relatively reliably.


Gamma-secretase inhibitor does not induce cytotoxicity in adult T-cell leukemia cell lines despite NOTCH1 expression.
Activated mutations in NOTCH1 are drivers of T-cell type acute lymphoblastic leukemia/lymphoma. The γ-secretase inhibitor (GSI), which suppresses the function of NOTCH1, is expected to be a molecular-targeted agent. NOTCH1 is also expressed in other malignant neoplasms. We aimed to determine the function of NOTCH1 expression and the effects of GSI on adult T-cell leukemia/lymphoma (ATL) caused by long-term human T-cell leukemia virus type I (HTLV-1) infection. We analyzed the expression of NOTCH1 in six ATL- and HTLV-1-infected cell lines and investigated the influence of activated NOTCH1 (i.e., the cleaved form of NOTCH1) together with GSI on cell proliferation. Activated NOTCH1 found in ATL- and HTLV-1-infected cell lines was undetectable after incubation with GSI, regardless of Tax expression (HTLV-1-coded protein). Whole-exome sequencing revealed that activated NOTCH1 mutations were undetectable in six ATL- and HTLV-1-infected cell lines, regardless of abundant NOTCH1 expression. Moreover, GSI did not suppress the growth of ATL cell lines. These findings suggested that NOTCH1 protein is constitutively activated but is likely a passenger during NOTCH1-mutation-negative ATL cell proliferation.


Changes in Local Public Health System Performance Before and After Attainment of National Accreditation Standards.
The aim of this study is to investigate the impact of Public Health Accreditation Board (PHAB) accreditation on the delivery of public health services and on participation from other sectors in the delivery of public health services in local public health systems. This study uses a longitudinal repeated measures design to identify differences between a cohort of public health systems containing PHAB-accredited local health departments and a cohort of public health systems containing unaccredited local health departments. It uses data spanning from 2006 to 2016. This study examines a cohort of local public health systems that serves large populations and contains unaccredited and PHAB-accredited local health departments. Data in this study were collected from the directors of health departments that include local public health systems followed in the National Longitudinal Study of Public Health Systems. The intervention examined is PHAB accreditation. The study focuses on 4 areas: the delivery of core public health services, local health department contribution toward these services, participation in the delivery of these services by other members of the public health system, and public health system makeup. Prior to the advent of accreditation, public health systems containing local health departments that were later accredited by PHAB appear quite similar to their unaccredited peers. Substantial differences between the 2 cohorts appear to manifest themselves after the advent of accreditation. Specifically, the accredited cohort seems to offer a broader array of public health services, involve more partners in the delivery of those services, and enjoy a higher percentage of comprehensive public health systems. The results of this study suggest that accreditation may yield significant benefits and may help public health systems develop the public health system capital necessary to protect and promote the public's health.


Genome sequence of the mesophilic Thermotogales bacterium Mesotoga prima MesG1.Ag.4.2 reveals the largest Thermotogales genome to date.
Here we describe the genome of Mesotoga prima MesG1.Ag4.2, the first genome of a mesophilic Thermotogales bacterium. Mesotoga prima was isolated from a polychlorinated biphenyl (PCB)-dechlorinating enrichment culture from Baltimore Harbor sediments. Its 2.97 Mb genome is considerably larger than any previously sequenced Thermotogales genomes, which range between 1.86 and 2.30 Mb. This larger size is due to both higher numbers of protein-coding genes and larger intergenic regions. In particular, the M. prima genome contains more genes for proteins involved in regulatory functions, for instance those involved in regulation of transcription. Together with its closest relative, Kosmotoga olearia, it also encodes different types of proteins involved in environmental and cell-cell interactions as compared with other Thermotogales bacteria. Amino acid composition analysis of M. prima proteins implies that this lineage has inhabited low-temperature environments for a long time. A large fraction of the M. prima genome has been acquired by lateral gene transfer (LGT): a DarkHorse analysis suggests that 766 (32%) of predicted protein-coding genes have been involved in LGT after Mesotoga diverged from the other Thermotogales lineages. A notable example of a lineage-specific LGT event is a reductive dehalogenase gene-a key enzyme in dehalorespiration, indicating M. prima may have a more active role in PCB dechlorination than was previously assumed.


Osseointegration of dental implants installed without mechanical engagement: a histometric analysis in dogs.
The purpose of this study was to elucidate the healing pattern of sand-blasted, large grid, acid-etched (SLA)-surfaced implants at two healing periods in a model that represents loosened implants (LIs) installed without mechanical engagement. Five mongrel dogs were used, in which 20 dental implants were prepared. The implants were divided into two groups according to the absence or presence of initial mechanical engagement: LIs) and control, respectively. An oversized drill was used to prepare the implant area for the LI group. The implants were allowed to heal for 4 or 8 weeks. After the healing period, the experimental animals were sacrificed and block sections were obtained for histological analysis and histometric measurements. All implants were in intimate contact with the host bone and were without any inflammation after both 4 and 8 weeks of healing. While the mean amount of bone-to-implant contact (BIC) was constant in the control group, it tended to increase in the LI group with increasing healing period. However, neither BIC nor bone density differed significantly between the groups or with the healing period. From the results of the study, it can be conjectured that the submerged and unloaded SLA-surfaced implants could result in successful osseointegration, even if the mechanical engagement was not obtained at placement of the implants.


Relationship between vision impairment and eye disease to vision-specific quality of life and function in rural India: the Aravind Comprehensive Eye Survey.
To determine the impact of vision impairment and eye diseases on vision-specific quality of life and visual function in an older population of rural southern India. Presenting and best-corrected visual acuity and burden of eye diseases were determined in a population aged 40 years and older, identified through a random cluster sampling strategy from 50 villages of rural south India. A questionnaire validated previously for use in this population was used to ascertain quality of life and visual function. Visual acuity measurements were obtained with illiterate E Early Treatment Diabetic Retinopathy Study (ETDRS) charts. Cataract was graded and defined based on the Lens Opacities Classification System (LOCS) III. Macular degeneration was defined based on the classification system proposed by the International ARM Epidemiologic Study Group. Glaucoma was defined based on results of clinical examinations including optic disc and visual fields. Analyses were performed to explore the relationship of overall and subscale quality-of-life and visual function scores with presenting acuity in the better-seeing eye, specific eye diseases, and demographic variables. Information on quality of life and visual function were available for 5119 (99.4%) of 5150 study subjects. The mean presenting visual acuity in the better eye was 0.76 +/- 0.53 logMAR (logarithm of the minimum angle of resolution) units. Age, education, occupation, presenting acuity in the better eye, and presence of a cataract, glaucoma, or refractive error were independently associated with overall quality-of-life and vision function scores. After adjustment for demographic variables and ocular disease, persons with vision impairment or bilateral blindness based on presenting visual acuity had lower scores across all domains of quality of life and vision function. Scores for subscales of quality-of-life and vision function domains were significantly lower among those with age-related cataract and glaucoma compared with persons without those eye diseases. Presenting vision in the better eye was associated with quality of life and vision function in this older population of rural south India. Subjects with glaucoma and age-related cataract had an associated decrease in quality of life and vision function, independent of presenting visual acuity in the better eye.


Retrospective Selection in Visual and Tactile Working Memory Is Mediated by Shared Control Mechanisms.
Selective attention regulates the activation of working memory (WM) representations. Retro-cues, presented after memory sample stimuli have been stored, modulate these activation states by triggering shifts of attention to task-relevant samples. Here, we investigated whether the control of such attention shifts is modality-specific or shared across sensory modalities. Participants memorized bilateral tactile and visual sample stimuli before an auditory retro-cue indicated which visual and tactile stimuli had to be retained. Critically, these cued samples were located on the same side or opposite sides, thus requiring spatially congruent or incongruent attention shifts in tactile and visual WM. To track the attentional selection of retro-cued samples, tactile and visual contralateral delay activities (tCDA and CDA components) were measured. Clear evidence for spatial synergy effects from attention shifts in visual WM on concurrent shifts in tactile WM were observed: Tactile WM performance was impaired, and tCDA components triggered by retro-cues were strongly attenuated on opposite-sides relative to same-side trials. These spatial congruency effects were eliminated when cued attention shifts in tactile WM occurred in the absence of simultaneous shifts within visual WM. Results show that, in contrast to other modality-specific aspects of WM control, concurrent attentional selection processes within tactile and visual WM are mediated by shared supramodal control processes.


A conservative triple antioxidant approach to the treatment of hepatitis C. Combination of alpha lipoic acid (thioctic acid), silymarin, and selenium: three case histories.
There has been an increase in the number of adults seeking liver transplantation for hepatitis C in the last few years and the count is going up rapidly. There is no reliable and effective therapy for chronic hepatitis C since interferon and antivirals work no more than 30% of the time, and liver transplant surgery is uncertain and tentative over the long run. This is because, ultimately, residual hepatitis C viremia infects the new liver. Furthermore, liver transplantation can be painful, disabling and extremely costly. The author describes a low cost and efficacious treatment program in 3 patients with cirrhosis, portal hypertension and esophageal varices secondary to chronic hepatitis C infection. This effective and conservative regimen combines 3 potent antioxidants (alpha-lipoic acid [thioctic acid], silymarin, and selenium) that possess antiviral, free radical quenching and immune boosting qualities. There are no remarkably effective treatments for chronic hepatitis C in general use. Interferon and antivirals have less than a 30% response rate and because of the residual viremia, a newly transplanted liver usually becomes infected again. The triple antioxidant combination of alpha-lipoic acid, silymarin and selenium was chosen for a conservative treatment of hepatitis C because these substances protect the liver from free radical damage, increase the levels of other fundamental antioxidants, and interfere with viral proliferation. The 3 patients presented in this paper followed the triple antioxidant program and recovered quickly and their laboratory values remarkably improved. Furthermore, liver transplantation was avoided and the patients are back at work, carrying out their normal activities, and feeling healthy. The author offers a more conservative approach to the treatment of hepatitis C, that is exceedingly less expensive. One year of the triple antioxidant therapy described in this paper costs less than $2,000, as compared to mor than $300,000 a year for liver transplant surgery. It appears reasonable, that prior to liver transplant surgery evaluation, or during the transplant evaluation process, the conservative triple antioxidant treatment approach should be considered. If these is a significant betterment in the patient's condition, liver transplant surgery may be avoided.


Targeting persisters for tuberculosis control.
Mycobacterial persisters, the survivors from antibiotic exposure, necessitate the lengthy treatment of tuberculosis (TB) and pose a significant challenge for our control of the disease. We suggest that persisters in TB are heterogeneous in nature and comprise various proportions of the population depending on the circumstances; the mechanisms of their formation are complex and may be related to those required for persistence in chronic infection. Results from recent studies implicate multiple pathways for persister formation, including energy production, the stringent response, global regulators, the trans-translation pathway, proteasomal protein degradation, toxin-antitoxin modules, and transporter or efflux mechanisms. A combination of specifically persister-targeted approaches, such as catching them when active and susceptible either by stimulating them to "wake up" or by intermittent drug dosing, the development of new drugs, the use of appropriate drug combinations, and combined chemotherapy and immunotherapy, may be needed for more effective elimination of persisters and better treatment of TB. Variations in levels of persister formation and in host genetics can play a role in the outcome of clinical treatment, and thus, these may entail personalized treatment regimens.


Nutritional intake and physical activity in leg ulcer patients.
The aim of the study was to describe the nutritional intake, meal patterns, physical activity and need for help in nine women living in their own homes and being treated for venous leg ulcers. Food habits were identified by use of interviews and food diaries completed by the women during a period of seven days. The intake of energy and nutrients from 304 eating events during seven days was calculated and meal patterns were evaluated using a qualitative system for meal classification. Physical activity and the degree of need were identified with the help of interviews. The intakes of energy and key nutrients for wound healing, such as protein, vitamin C and zinc, were not optimal according to the Swedish nutrition recommendations, although food habits were well organized. Most of the women had hardly any physical activities and the need of help and support varied, from daily visits to visits every second week.


Co-LncRNA: investigating the lncRNA combinatorial effects in GO annotations and KEGG pathways based on human RNA-Seq data.
Long non-coding RNAs (lncRNAs) are emerging as key regulators of diverse biological processes and diseases. However, the combinatorial effects of these molecules in a specific biological function are poorly understood. Identifying co-expressed protein-coding genes of lncRNAs would provide ample insight into lncRNA functions. To facilitate such an effort, we have developed Co-LncRNA, which is a web-based computational tool that allows users to identify GO annotations and KEGG pathways that may be affected by co-expressed protein-coding genes of a single or multiple lncRNAs. LncRNA co-expressed protein-coding genes were first identified in publicly available human RNA-Seq datasets, including 241 datasets across 6560 total individuals representing 28 tissue types/cell lines. Then, the lncRNA combinatorial effects in a given GO annotations or KEGG pathways are taken into account by the simultaneous analysis of multiple lncRNAs in user-selected individual or multiple datasets, which is realized by enrichment analysis. In addition, this software provides a graphical overview of pathways that are modulated by lncRNAs, as well as a specific tool to display the relevant networks between lncRNAs and their co-expressed protein-coding genes. Co-LncRNA also supports users in uploading their own lncRNA and protein-coding gene expression profiles to investigate the lncRNA combinatorial effects. It will be continuously updated with more human RNA-Seq datasets on an annual basis. Taken together, Co-LncRNA provides a web-based application for investigating lncRNA combinatorial effects, which could shed light on their biological roles and could be a valuable resource for this community. Database URL: http://www.bio-bigdata.com/Co-LncRNA/.


Modifying excitation transfer cross sections with an ac Stark effect.
We show that it is possible to manipulate electronic energy transfer collision cross sections between different atomic species by using a strong electromagnetic field close to resonance with a transition between two excited states to modify the energy levels (i.e., to create dressed states), which may be placed in or out of resonance with populated states (forming a population reservoir) in one of the species. We outline an estimate for a transfer cross section for a demonstration scheme and show that cross-section enhancements up to the order of 10(3) are possible.


Characterization of traffic-related PM concentration distribution and fluctuation patterns in near-highway urban residential street canyons.
Analyses of outdoor traffic-related particulate matter (PM) concentration distribution and fluctuation patterns in urban street canyons within a microscale distance of less than 500 m from a highway source are presented as part of the results from the Brooklyn Traffic Real-Time Ambient Pollutant Penetration and Environmental Dispersion (B-TRAPPED) study. Various patterns of spatial and temporal changes in the street canyon PM concentrations were investigated using time-series data of real-time PM concentrations measured during multiple monitoring periods. Concurrent time-series data of local street canyon wind conditions and wind data from the John F. Kennedy (JFK) International Airport National Weather Service (NWS) were used to characterize the effects of various wind conditions on the behavior of street canyon PM concentrations.Our results suggest that wind direction may strongly influence time-averaged mean PM concentration distribution patterns in near-highway urban street canyons. The rooftop-level wind speeds were found to be strongly correlated with the PM concentration fluctuation intensities in the middle sections of the street blocks. The ambient turbulence generated by shifting local wind directions (angles) showed a good correlation with the PM concentration fluctuation intensities along the entire distance of the first and second street blocks only when the wind angle standard deviations were larger than 30 degrees. Within-canyon turbulent shearing, caused by fluctuating local street canyon wind speeds, showed no correlation with PM concentration fluctuation intensities. The time-averaged mean PM concentration distribution along the longitudinal distances of the street blocks when wind direction was mostly constantly parallel to the street was found to be similar to the distribution pattern for the entire monitoring period when wind direction fluctuated wildly. Finally, we showed that two different PM concentration metrics-time-averaged mean concentration and number of concentration peaks above a certain threshold level-can possibly lead to different assessments of spatial concentration distribution patterns.


Cerebrospinal fluid secretory Ca2+-dependent phospholipase A2 activity is increased in Alzheimer disease.
The phospholipase A(2) (PLA2) family comprises multiple isoenzymes that vary in their physicochemical properties, cellular localizations, calcium sensitivities, and substrate specificities. Despite these differences, PLA2s share the ability to catalyze the synthesis of the precursors of the proinflammatory mediators. To investigate the potential of PLA2 as a biomarker in screening neuroinflammatory disorders in both clinical and research settings, we developed a PLA2 assay and determined the predominant types of PLA2 activity in cerebrospinal fluid (CSF). We used liposomes composed of a fluorescent probe (bis-Bodipy FL C11-PC [1,2-bis-(4,4- difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-undecanoyl)-sn-glycero-3-phosphocholine]) and 1,2-dioleoyl-l-alpha-phosphatidylcholine as a substrate to measure CSF PLA2 activity in a 96-well microtiter plate format. We established the type of CSF PLA2 activity using type-specific inhibitors of PLA2. Using 5 microL CSF per assay, our PLA2 activity assay was reproducible with CVs <15% in 2 CSF samples and for recombinant secretory Ca(2+)-dependent PLA2 (sPLA2) in concentrations ranging from 0.25 to 1 micromol/L. This PLA2 assay allowed identification of sPLA2 activity in lumbar CSF from healthy individuals 20-77 years old that did not depend on either sex or age. Additionally, CSF sPLA2 activity was found to be increased (P = 0.0008) in patients with Alzheimer disease. Adult human CSF has sPLA2 activity that can be measured reliably with the assay described. This enzyme activity in the CSF is independent of both sex and age and might serve as a valuable biomarker of neuroinflammation, as we demonstrated in Alzheimer disease.


An engineered pancreatic cancer model with intra-tumoral heterogeneity of driver mutations.
Pancreatic ductal adenocarcinoma (PDAC) is a complex disease with significant intra-tumoral heterogeneity (ITH). Currently, no reliable PDAC tumor model is available that can present ITH profiles in a controlled manner. We develop an in vitro microfluidic tumor model mimicking the heterogeneous accumulation of key driver mutations of human PDAC using cancer cells derived from genetically engineered mouse models. These murine pancreatic cancer cell lines have KPC (Kras and Trp53 mutations) and KIC genotypes (Kras mutation and Cdkn2a deletion). Also, the KIC genotypes have two distinct phenotypes - mesenchymal or epithelial. The tumor model mimics the ITH of human PDAC to study the effects of ITH on the gemcitabine response. The results show gemcitabine resistance induced by ITH. Remarkably, it shows that cancer cell-cell interactions induce the gemcitabine resistance potentially through epithelial-mesenchymal-transition. The tumor model can provide a useful testbed to study interaction mechanisms between heterogeneous cancer cell subpopulations.


Update on therapeutic developments for hereditary angioedema.
Remarkable progress has been made in understanding the molecular mechanisms underlying attacks of swelling in hereditary angioedema (HAE). Treatment options in the United States for this potentially life-threatening disease had remained essentially static, however, over the past 40 years. Prophylactic therapy had relied on attenuated androgens or antifibrinolytic agents. Although demonstrably effective, these drugs have been fraught with side effects. Acute therapy has been largely relegated to supportive care. In this article we discuss emerging treatments that have evolved from the recognition that kinin generation is the fundamental abnormality leading to attacks of angioedema. We will review the newly approved replacement therapy for prophylaxis of HAE attacks with C1 inhibitor (C1INH). Potential options for the acute treatment of HAE will be discussed including purified C1INH, recombinant C1INH, an inhibitor of plasma kallikrein, and a B2-receptor antagonist. The arrival of these novel therapies promises to transform the future management of HAE.


The role of staff in health promotion in community residences for people with intellectual disabilities: variation in views among managers and caregivers.
Managers and caregivers in community residences for people with intellectual disabilities are expected both to promote residents' health and to support their autonomy. The aim of this article was to explore variation in views among managers and caregivers on the role of staff in health promotion. A qualitative study was conducted using semi-structured interviews with six managers and six caregivers. The analysis used a phenomenographic approach to categorize variation in views. We identified five qualitatively different main categories of roles staff play in health promotion: the parent, the manipulator, the coach, the educator and the libertarian. In addition lifestyle-related risk factors for ill-health and barriers to a healthy lifestyle were analysed and described using qualitative content analysis. The results highlight the ethical conflict that faces staff trying to support a healthy lifestyle as well as the autonomy of the residents.


PrP genotypes of atypical scrapie cases in Great Britain.
Great Britain and elsewhere have detected atypical scrapie infection in sheep with PrP genotypes thought to be genetically resistant to the classical form of scrapie. DNA sequencing of the PrP gene of British atypical scrapie cases (n=69), classical scrapie cases (n=59) and scrapie-free controls (n=138) was undertaken to identify whether PrP variants, other than the three well-characterized polymorphic codons, influenced susceptibility to atypical scrapie infection. Four non-synonymous changes, M112T, M137T, L141F and P241S, were detected that are most probably associated with the A(136)R(154)Q(171) haplotype. Only the PrP variant containing a phenylalanine residue at amino acid position 141 was found to be associated more commonly with the atypical scrapie cases. In addition to the single nucleotide polymorphisms associated with the ARQ allele, two out of nine atypical scrapie cases with the ARR/ARR genotype were found to contain a 24 bp insertion, leading to an additional octapeptide repeat. In terms of PrP genetics, one classification of the GB scrapie cases examined in this study would place animals carrying any homozygous or heterozygous combination of ARR, AHQ or AF(141)RQ alleles, or any one of these alleles when paired with ARQ, as being susceptible to atypical scrapie infection, and animals heterozygous or homozygous for VRQ or homozygous for ARQ as being susceptible to classical scrapie disease. The AHQ PrP allele was associated with the highest incidence of atypical scrapie (263 per 100 000 alleles), whilst VRQ was associated with the lowest incidence (10 per 100 000 alleles).


Incidence rates of admissions associated with alcohol withdrawal syndrome in Spain: Analysis of minimum basic data set 1999-2010.
There are no data on the incidence of admissions associated with alcohol withdrawal syndrome (AWS) or about its trend over time in Spain. To analyze the characteristics, incidence rates and trends over time of hospital admissions associated with AWS in Spanish public hospitals. Analysis from the Spanish public hospitals minimum basic data set of hospital admissions with AWS (CIE9-MC 291.81), alcohol withdrawal delirium (CIE9-MC 291.0) and alcohol withdrawal hallucinosis (CIE9-MC 291.3), since 1999 to 2010. We identified 56,395 admissions associated with AWS. Mean age was 50.9 (SD 12.5) and 88% were male. The most frequent admission department was Internal Medicine (24.9%). The mean hospital stay was 12.6 days (SD 14.4) and mortality was 4.7%; 62.6% of cases developed AWS during an admission for another reason, mostly due to alcohol-related pathologies. Secondary diagnoses in patients hospitalized for AWS were related to alcohol consumption in more than half of the cases. The incidence rate of admissions associated with AWS in Spain remained stable from 1999 to 2010, with a small decline in the last 3 years of the period. The communities with the highest incidence were the Canary Islands, the Balearic Islands and Galicia. The incidence rate of admissions associated with AWS in Spanish public hospitals in the period 1999-2010 has remained stable with slight changes. There are differences in the incidence of AWS among the different autonomous communities.


Estimation of 10-Year Risk of Death from Coronary Heart Disease, Stroke, and Cardiovascular Disease in a Pooled Analysis of Japanese Cohorts: EPOCH-JAPAN.
We aimed to develop and validate risk prediction models to estimate the absolute 10-year risk of death from coronary heart disease (CHD), stroke, and cardiovascular disease (CVD). We evaluated a total of 44,869 individuals aged 40-79 years from eight Japanese prospective cohorts to derive coefficients of risk equations using cohort-stratified Cox proportional hazard regression models. Discrimination (C-index) of the equation was examined in each cohort and summarised using random-effect meta-analyses. Calibration of the equation was assessed using Hosmer-Lemeshow chi-squared statistic. Within a median follow-up of 12.7 years, we observed 765 deaths due to CVD (276 CHDs and 489 strokes). After backward selection, age, sex, current smoking, systolic blood pressure (SBP), proteinuria, prevalent diabetes mellitus, the ratio of total cholesterol to high-density lipoprotein cholesterol (TC/HDLC), interaction terms of age by SBP, and age by current smoking were retained as predictors for CHD. Sex was excluded in the stroke equation. We did not consider TC/HDLC as a risk factor for the stroke and CVD equations. The pooled C-indices for CHD, stroke, and CVD were 0.83, 0.80, and 0.81, respectively, and the corresponding p-values of the Hosmer-Lemeshow tests were 0.18, 0.003, and 0.25, respectively. Risk equations in the present study can adequately estimate the absolute 10-year risk of death from CHD, stroke, and CVD. Future work will evaluate the system as an education and risk communication tool for primary prevention of CHD and stroke.


[Genetics of osteoporosis].
Osteoporosis is a common disease with a strong genetic componenet characterized by reduced bone mass and increased risk of fragility fractures. Twins and family studies have shown that the heritability of bone mineral density (BMD) and other determinants of fracture risk - such as ultrasound properties of bone, skeletal geometry, and bone turnover - are high, although heritability of fracture is modest. In this article, some important gene identifications of the last years such as vitamin receptor, estrogen receptor alpha, alpha 1 chain of type I collagen, low density lipoprotein receptor - related protein 5, bone morphogenic proteins 2 and 4, transforming growth factor beta 1, receptor activator of NF-kappa-B, receptor activator of NF-kappa-B ligand, osteoprotegerin and their contribution to the understanding of the genetic regulation of bone mass and susceptibility to osteoporosis are present.


Specific PCR assays for the detection and quantification of DNA from the biocontrol strain Trichoderma harzianum 2413 in soil.
Strain identification in situ is an important factor in the monitoring of microorganisms used in the field. In this study, we demonstrated the use of sequence-characterized amplified region (SCAR) markers to detect genomic DNA from Trichoderma harzianum 2413 from soil. Two primers (SCAR A1/SCAR A1c) were tested against DNA of 27 isolates of Trichoderma spp. and amplified a 990-bp fragment from T. atroviride 11 and a 1.5-kb fragment from T. harzianum 2413, using an annealing temperature of 68 degrees C. These fragments showed no significant homology to any sequence deposited in the databases. The primer pair, BR1 and BR2, was designed to the 1.5-kb fragment amplified from T. harzianum 2413, generating a SCAR marker. To test the specificity of these primers, experiments were conducted using the DNA from 27 Trichoderma spp. strains and 22 field soil samples obtained from four different countries. PCR results showed that BR1 and BR2 amplified an 837-bp fragment unique to T. harzianum 2413. Assays in which total DNA was extracted from sterile and nonsterile soil samples, inoculated with spore or mycelium combinations of Trichoderma spp. strains, indicated that the BR1 and BR2 primers could specifically detect T. harzianum 2413 in a pool of mixed DNA. No other soil-microorganisms containing these sequences were amplified using these primers. To test whether the 837-bp SCAR marker of T. harzianum 2413 could be used in real-time PCR experiments, new primers (Q2413f and Q2413r) conjugated with a TaqMan fluorogenic probe were designed. Real-time PCR assays were applied using DNA from sterile and nonsterile soil samples inoculated with a known quantity of spores of Trichoderma spp. strains.


Effect of social organisation on interspecific differences in overmarking behaviour of foals in African equids.
Overmarking remains an unstudied topic in juvenile mammals. We have previously documented a very high rate of overmarking by foals in four captive African equid species: mountain zebra (Equus zebra), plains zebra (Equus quagga), Grévy's zebra (Equus grevyi), and African wild ass (Equus africanus). African equids vary interspecifically in their social organisation. Since differences in social organisation affect many mammalian behaviours, in this study we investigated interspecific differences in overmarking behaviour of foals, analysing only cases where elimination of any other individual was explored by a foal. We hypothesised that the pattern of overmarking by foals should reflect either differences in social organisation of the species or phylogenetic relations among them. We found that in all species very young foals explored mostly maternal eliminations, and this preference declined with increasing age of the foal and reflected the social organisation of the species; the highest overmarking rate was in species with high intragroup aggression (mountain zebra) and lowest in species with low intragroup aggression and which form crèches (African wild ass). Similarly, the rate of overmarking of the mother, as opposed to other herdmates, was associated with social organisation of the respective species. Thus, we found interspecific differences in overmarking by foals, which were associated with variability in social organisation. Since we also revealed differences between African wild ass and zebra behaviour in early stages of ontogeny, we cannot refute the effect of phylogeny on overmarking behaviour. Additionally, our results supported the identity sharing hypothesis as an explanation of overmarking.


Monoclonal antibodies for the treatment of refractory asthma.
A small proportion of patients with asthma have severe disease characterized by persistent airflow obstruction, airway hyperresponsiveness and eosinophilic airway inflammation. This review focuses on the clinical efficacy of inhibiting T helper 2-cytokine-mediated inflammatory responses using monoclonal antibodies directed against immunoglobulin E (IgE), interleukin (IL)-5, and IL-4/IL-13 in patients with severe refractory asthma. The heterogeneity of airway inflammation in severe asthma has led to the recognition of multiple pathophysiologically distinct severe asthma endotypes. Biomarkers are being developed and evaluated to identify these endotypes and to guide the use of specific biologics in the appropriate patients who remain uncontrolled on high doses of inhaled corticosteroids and long-acting bronchodilators or oral corticosteroids. Examples include the efficacy of omalizumab in patients with severe refractory atopic asthma characterized by raised serum total IgE, mepolizumab, reslizumab, and benralizumab in patients with recurrent eosinophilic exacerbations characterized by blood and sputum eosinophilia despite high doses of corticosteroids, and lebrikizumab, pitrakinra, dupilumab, and tralokinumab that target the IL-4/IL-13 signalling pathways in patients with eosinophilic asthma or raised serum periostin. In severe refractory asthma, both an understanding of the underlying pathophysiologic mechanisms driving airway inflammation and the identification of appropriate biomarkers in individual patients are critical in guiding the use of biologics and monoclonal antibodies that target the specific pathological processes.


Myoclonus from selective dentate nucleus degeneration in type 3 Gaucher disease.
To describe a case with a new genetic variant of type 3 Gaucher disease presenting with stimulus-sensitive and action myoclonus in the presence of selective dentate abnormalities. Clinical, pathologic, and molecular genetic studies. Medical school departments. A 6-year-old girl with type 3 Gaucher disease experienced progressively crippling generalized stimulus-sensitive and action myoclonus. Repeated electroencephalographic examination did not show cortical activity associated with the myoclonus, suggesting its subcortical origin. Neuropathological examination revealed selective degeneration of the cerebellar dentate nucleus and dentatorubrothalamic pathway in the face of essentially complete lack of storage in the brain. Mutation analysis identified the following 2 mutant alleles: one with a V394L mutation and the other with the lesion RecTL (D409H + L444P + A456P + V460V), which resulted from a recombination event, with the pseudogene located 16 kilobases downstream from the structural gene. Given the restricted abnormalities, this genetically unique case provides insight into the pathogenesis of myoclonus and suggests a prominent role for the cerebellar dentate nucleus in its genesis.


Temporally multiplexed imaging of dynamic signaling networks in living cells.
Molecular signals interact in networks to mediate biological processes. To analyze these networks, it would be useful to image many signals at once, in the same living cell, using standard microscopes and genetically encoded fluorescent reporters. Here, we report temporally multiplexed imaging (TMI), which uses genetically encoded fluorescent proteins with different clocklike properties-such as reversibly photoswitchable fluorescent proteins with different switching kinetics-to represent different cellular signals. We linearly decompose a brief (few-second-long) trace of the fluorescence fluctuations, at each point in a cell, into a weighted sum of the traces exhibited by each fluorophore expressed in the cell. The weights then represent the signal amplitudes. We use TMI to analyze relationships between different kinase activities in individual cells, as well as between different cell-cycle signals, pointing toward broad utility throughout biology in the analysis of signal transduction cascades in living systems.


Thermal properties measurement and laser operation of a new Yb: Gd0.68Lu0.32VO4 crystal.
A new series mixed vanadate crystals, Yb:Gd(x)Lu(1-x)VO(4) (x=0.2, 0.35, 0.53, 0.68 and 0.84), were grown by the Czochralski method. The thermal conductivities of these crystals were measured from room temperature to 280?C, and the thermal conductivities of Yb:Gd(0.68)Lu(0.32)VO(4) at room temperature are 6.4 and 6.9 W m(-1)K(-1) along a- and c-axis, respectively. The material constants of Yb:Gd(x)Lu(1-x)VO(4) series crystals were estimated. The properties of Yb:Gd(x)Lu(1-x)VO(4) series crystals were compared with those of Yb:LuVO(4) and Yb:GdVO(4) crystals. Laser output power of 1.25 W at 1031 nm was obtained with a slope efficiency of 26% by use of diode pumping for Yb:Gd(0.68)Lu(0.32)VO(4) crystal.


Structural racism, economic opportunity and racial health disparities: Evidence from U.S. counties.
In this study, we introduce the 'racial opportunity gap' as a place-based measure of structural racism for use in population health research. We first detail constructing the opportunity gap using race-sex specific estimates of intergenerational economic mobility outcomes for a recent cohort. We then illustrate its utility in examining spatial variation in the racial mortality gap. First we demonstrate a correlation between the racial opportunity gap and the racial mortality gap across U.S. counties; where the gap in the adult earnings of black and white children born to families at the same income level is greater so, too, is the gap in mortality. Second, we show in a multivariable framework that the racial opportunity gap is associated with the racial mortality gap net of differences in the socioeconomic composition of the two groups. In so doing, we aim to provide population health researchers with a new empirical tool and analytic framework for examining the role of structural racism in generating racial health disparities.


Antibody titres against canine papillomavirus 1 peak around clinical regression in naturally occurring oral papillomatosis.
Most forms of canine papillomatosis are believed to be associated with papillomavirus infections. Canine papillomavirus type 1 (CPV1) is considered to be responsible for most oral cases and several forms of cutaneous papillomatosis. The aim of this study was to evaluate cases of naturally occurring oral papillomatosis with regard to the type of virus involved, antibody induction and remission time. Forty dogs showing different degrees of classical oral papillomatosis were included as a single study group. Tissue and serum samples were acquired upon initial presentation; serum samples were collected again upon remission (n = 13) and after 3 months of convalescence (n = 4). None of the dogs underwent antiviral therapy. Tissue samples were tested by PCR to detect CPV DNA, while serum samples were tested using a specific enzyme-linked immunosorbent assay for antibodies against the L1 capsid protein of CPV1. All tissue samples were positive for CPV1 DNA, and 87.5% of all serum samples contained measurable levels of antibody against the virus (cut-off value 0.3). The average optical density measured in the enzyme-linked immunosorbent assay was 0.51 at initial presentation, 1.65 upon remission and 0.83 at 3 months postrecovery. Time to clinical regression varied between 1 month and 1 year. These data support existing evidence for a high prevalence of CPV1 in canine oral papillomatosis. The healing process seems to correlate with a strong antibody response, and antibody titres peaked around the time of clinical recovery. In contrast to previous data from laboratory settings, the variation in remission time was very high.


Partial purification and characterization of heat-stable enterotoxin produced by Yersinia enterocolitica.
By using a suckling mouse assay, heat-stable enterotoxin (ST) was purified from the culture filtrate of Yersinia enterocolitica isolated from a diarrheal patient. The purification procedures involve ultrafiltration with an Amicon HIP-10 hollow fiber, ethanol fractionation, protamine sulfate treatment, diethylaminoethyl-Sephacel and hydroxylapatite column chromatographies, and Sephacryl S-200 superfine gel filtration. About 408-fold purification was achieved, with a yield of 12.0%. The minimal effective dose of purified ST was about 110 ng in the suckling mouse assay. The molecular weight of purified ST was 9,000 by Sephadex G-100 superfine gel filtration. The purified ST was stable to heating (100 degrees C for 20 min, 121 degrees C for 20 min) and did not lose its toxicity after treatment with protease, trypsin, lipase, phospholipase C, ribonuclease, deoxyribonuclease, beta-glucosidase, and neuraminidase. The purified ST was separated by isoelectric focusing into two active fractions, with pI's of 3.29 (ST-1) and 3.00 (ST-2), respectively. Antiserum from guinea pigs immunized with the purified ST neutralized the activity of both Y. enterocolitica ST and Escherichia coli ST.


Biallelic inactivation of the thyroid hormone receptor beta1 gene in early stage breast cancer.
Loss of heterozygosity within the short arm of chromosome 3 is a common molecular event in several types of solid tumors. In breast cancer, 3p loss of heterozygosity occurs in invasive tumor cells as well as in morphologically normal terminal ductal lobular units adjacent to carcinoma in some cases [G. Deng et al., Science (Wash. DC), 274: 2057-2059, 1996.]. The most frequent region of allelic loss at 3p24.3 in morphologically normal terminal ductal lobular units encompasses the thyroid hormone receptor beta1 (TRbeta1) gene. Here we have observed a variable degree of TRbeta1 promoter hypermethylation in all 11 cases of primary breast cancer examined. Moreover, hypermethylation occurred at the same CpG sites in nonmalignant tissue peripheral to carcinoma in 4 of 11 cases. The lack of TRbeta1 nuclear staining, a likely result of biallelic gene inactivation, was observed in 25% (22 of 85) of primary tumors. This is a first demonstration of promoter hypermethylation and a concurrent reduction of TRbeta1 transcripts in breast cancer cell lines, although specific CpG sites targeted for gene silencing remain to be determined. Gene expression was restored by treatment with 5-aza-deoxycytidine in such cases. The observation of early, frequent, and multiple mechanisms of TRbeta1 inactivation suggests a potential role for this gene in the suppression of breast tumorigenesis.


Color accuracy of resin cements and try-in pastes.
The color accuracy of resin composite cements and their corresponding try-in pastes was assessed. Three shades of three brands were compared using the cements alone and in contact with a standard porcelain disk. There were significant differences in color between resin cements and their corresponding try-in pastes. The addition of a 1-mm-thick porcelain disk greatly reduced the apparent color differences. The color of resin cements changed over time, but not significantly. Baseline measurements of resin cement colors were compared and measurements made after 24 hours of storage. Color changes were statistically significant but not perceptible, and all samples became slightly darker with age.


Artificial sunlight irradiation induces ultraweak photon emission in human skin fibroblasts.
Photons participate in many atomic and molecular interactions and changes in the physical universe. In recent years sophisticated detection procedures for the measurement of ultraweak photons in a variety of different cells have been performed leading to the conclusion that plant, animal and human cells emit ultraweak photons. Using an extremely low-noise, high-sensitive photon-counting system, which allows maximal exploitation of the potential capabilities of a photomultiplier tube, ultraweak photons were quantitated in human skin fibroblasts. It was found that light from an artificial sunlight source induces ultraweak photon emission in these cells. However, the results demonstrate that this induction is significantly lower in normal fibroblasts compared with those obtained from a donor suffering from xeroderma pigmentosum disease group A, a disease characterized by deficient repair of DNA. The largest increase in ultraweak photon emission after UV exposure was measured in mitomycin-C-induced post-mitotic xeroderma pigmentosum cells which showed 10-20 times higher ultraweak photon intensities than mitotic UV-irradiated normal cells. These data suggest that xeroderma pigmentosum cells tend to lose the capacity of efficient storage of ultraweak photons, indicating the existence of an efficient intracellular photon trapping system within human cells.


Bilateral tarsal tunnel syndrome attributed to bilateral fibrous tarsal coalition and symmetrical hypertrophy of the sustentaculum tali.
Tarsal tunnel syndrome (TTS) is one of the frequently encountered entrapment neuropathies of the lower limb. Most often the aetiology is considered to be idiopathic. However, it has to be thoroughly investigated to rule out diverse extrinsic and intrinsic causes. The sustentaculum tali (ST) forms the superior part of the distal tarsal tunnel, and any increase in its size may result in irritation of the tibial nerve or its branches, the medial and lateral plantar nerves or decrease the overall volume of the tarsal tunnel culminating to clinical features of TTS. A hypertrophied ST is an extremely rare cause for TTS, and we report perhaps a first case of bilateral TTS in a young adult due to bilateral symmetrical hypertrophied ST which in turn was a result of bilateral fibrous tarsal coalition. Staged bilateral surgical excision of the hypertrophied part of ST and the underlying fibrous tarsal coalition ensured prompt symptomatic relief.


Survival of three species of anuran metamorphs exposed to UV-B radiation and the pathogenic fungus Batrachochytrium dendrobatidis.
When exploring the possible factors contributing to population declines, it is necessary to consider multiple, interacting environmental stressors. Here, we investigate the impact of 2 factors, ultraviolet radiation and disease, on the survival of anuran amphibians. Exposure to ultraviolet-B (UV-B) radiation increases mortality and results in various sub-lethal effects for many amphibian species. Infectious diseases can also negatively impact amphibian populations. In this study, we exposed metamorphic individuals (metamorphs) to both UV-B and Batrachochytrium dendrobatidis (BD), a fungal pathogen and cause of the disease chytridiomycosis, and monitored survival for 3 wk. We tested for possible interactions between UV-B and BD in 3 species: the Cascades frog Rana cascadae; the Western toad Bufo boreas; and the Pacific treefrog Hyla regilla. We found strong interspecific differences in susceptibility to BD. For example, R. cascadae suffered a large increase in mortality when exposed to BD; B. boreas also experienced mortality, but this effect was small relative to the R. cascadae response. H. regilla did not show any decrease in survival when exposed to either factor. No synergistic interactions between UV-B and BD were found for any of the test species. A previous study investigating the impact of BD on larval amphibians showed different species responses (Blaustein et al. 2005a). Our results highlight the importance of studying multiple life history stages when determining the impact of environmental stressors. The contrast between these 2 studies emphasizes how vulnerability to a pathogen can vary between life history stages within a single species.


Counting unstained, confluent cells by modified bright-field microscopy.
We present a very simple procedure yielding high-contrast images of adherent, confluent cells such as human neuroblastoma (SH-EP) cells by ordinary bright-field microscopy. Cells are illuminated through a color filter and a pinhole aperture placed between the condenser and the cell culture surface. Refraction by each cell body generates a sharp, bright spot when the image is defocused. The technique allows robust, automatic cell counting from a single bright-field image in a wide range of focal positions using free, readily available image-analysis tools. Contrast may be enhanced by swelling cell bodies with a brief incubation in PBS. The procedure was benchmarked against manual and automated counting of fluorescently labeled cell nuclei. Counts from day-old and freshly seeded plates were compared in a range of densities, from sparse to densely overgrown. On average, bright-field images produced the same counts as fluorescence images, with less than 5% error. This method will allow routine cell counting using a plain bright-field microscope without cell-line modification or cell staining.


Pedunculopontine nucleus stimulation improves gait freezing in Parkinson disease.
Pedunculopontine nucleus (PPN) stimulation is a novel therapy for Parkinson disease. However, controversies remain regarding the clinical application of this new therapy, including patient selection, electrode positioning, and how best to assess outcomes. To clarify the clinical application of PPN stimulation in Parkinson disease. Five consecutive patients with Parkinson disease complicated by severe gait freezing, postural instability, and frequent falls (all persisting even while the patient was on medication) received bilateral stimulation of the mid-lower PPN without costimulation of other brain targets. Outcomes were assessed prospectively over 2 years with gait-specific questionnaires and the Unified Parkinson Disease Rating Scale (part III). The primary outcome, the Gait and Falls Questionnaire score, improved significantly with stimulation. Benefits were maintained over 2 years. Unified Parkinson Disease Rating Scale (part III) items assessing gait and posture were relatively insensitive to these treatment effects. Beneficial effects often appeared to outlast stimulation for hours or longer. Thus, single-session on- vs off-stimulation assessments may be susceptible to "delayed washout effects." Stimulation of the PPN did not change akinesia scores or dopaminergic medication requirements. Bilateral stimulation of the mid-lower PPN (more caudal than previous reports) without costimulation of other brain targets may be beneficial for the subgroup of patients with Parkinson disease who experience severe gait freezing and postural instability with frequent falls, which persist even while on medication. Choosing appropriate outcome measures and accounting for the possibility of prolonged stimulation washout effects appear to be important for detecting the clinical benefits.


Preliminary crystallographic data for NAD(P)H quinone reductase isolated from the Walker 256 rat carcinoma cell line.
An NAD(P)H quinone reductase isolated from Walker rat 256 carcinoma cells has been crystallized in a form suitable for high-resolution structural analysis. The crystals belong to orthorhombic space group P2(1)2(1)2(1) with cell parameters a = 168.15 A, b = 105.09 A and c = 67.38 A and contain four monomeric or two dimeric enzyme molecules per asymmetric unit. Diffraction extends beyond 2.3 A resolution.


Saphenofemoral vein crossover bypass grafting in iliofemoral vein obstruction.
Iliofemoral vein obstruction that causes venous hypertension in an extremity may produce symptoms that severely limit a patient's activities. A saphenofemoral venous crossover graft may relieve those symptoms. The procedure also is indicated for the repair of the venous system following severe, limb-threatening vascular trauma. For chronic obstruction, the procedure should be delayed for six months to allow for the possible development of adequate collateral circulation.


[Ginsenoside Rh₂-induced inhibition of histone deacetylase 6 promotes K562 cells autophagy and apoptosis in vivo].
To study the in vivo inhibition effect of ginsenoside Rh₂ on humanleukemia cells, and explore its mechanism from autophagy and apoptosis aspects, human leukemia K562 cells allograft tumor models were applied, and after administration of ginsenosides Rh₂ by gavage, the tumor diameter, volume and inhibitory rate were measured, and the anti-tumor activity of ginsenosides Rh₂ was observed. The levels of HAT and HDAC in tumor tissues were detected by chemical colorimetry assay, and expressions of HDAC1, HDAC2, HDAC3, HDAC4, HDAC5 and HDAC6 were detected by Western blotting assay. The expression levels of vital genes closely associated with autophagy and mRNA expressions of HDAC6 and Hsp90 were detected by Real time-PCR. HE staining was used to observe apoptosis, and immunohistochemistry was used to detect the protein expressions of HDAC6, Hsp90 and activated caspases 3. The results showed that ginsenoside Rh₂ could inhibit the growth of k562 cells allograft tumor, with a tumor inhibition rate up to 53.10%. Ginsenoside Rh₂ could significantly decrease HDAC activity and decrease the expressions of HDAC1, HDAC2 and HDAC6, and inhibit the expressions of HDAC6 and HSP90, increase the expressions of vital autophagy genes (beclin-1, LC3A and LC3B). Histopathological results showed that ginsenosides Rh₂ could significantly increase the tumor apoptosis. Therefore, ginsenoside Rh₂ had good anti-tumor effect in vivo, and the mechanism maybe associated with regulating autophagy and apoptosis through HDAC6 and Hsp90 pathways and inhibiting the in vivo proliferation of tumor cells.


Trends in the antimicrobial susceptibilities and serotypes of Streptococcus pneumoniae: results from the Tigecycline In Vitro Surveillance in Taiwan (TIST) study, 2006-2010.
Isolates of Streptococcus pneumoniae (n = 530) were collected from 20 hospitals in different parts of Taiwan from 2006 to 2010. MICs to 16 antimicrobial agents were determined by broth dilution method and serotypes were identified by latex agglutination. Based on meningitis (non-meningitis) criteria established by the CLSI, 11.7% (63.2%) of all isolates were susceptible to penicillin and 46.0% (83.8%) were susceptible to ceftriaxone. Of the isolates, 94.3% were non-susceptible to azithromycin and 5.8% and 7.2% were non-susceptible to moxifloxacin and levofloxacin, respectively. Susceptibility to penicillin by meningitis criteria increased significantly (P = 0.0012) with year, and that to clindamycin and amoxicillin/clavulanic acid declined significantly (P < 0.05). Six major serotypes were found, namely 19F (24.0%), 23F (18.5%), 14 (13.6%), 6B (12.5%), 19A (7.5%) and 3 (5.1%). Prevalence of serotypes 19F and 14 remained stationary, that of serotype 6B decreased significantly (P < 0.0001) and that of serotype 19A increased significantly (P < 0.0001) with year. The coverage rate of PCV-7 among the pneumococcal isolates declined from 80.5% in 2006 to 50% in 2010 (P < 0.0001) and that of PCV-13 declined from 91.5% in 2009 to 75% in 2010. The non-susceptibility rate to levofloxacin was highest among serotype 23F isolates (13.3%) and lowest among serotype 19A isolates (2.5%). Rates of resistance to the four agents penicillin, ceftriaxone, azithromycin and clindamycin were highest among serotype 19A isolates (70.0%) and 23F isolates (49.0%). All serotype 3 isolates were susceptible to four of the most commonly used antibiotics (penicillin, ceftriaxone, azithromycin and levofloxacin).


Systemic up-regulation of angiotensin II type 1 receptor in cardiac donors with spontaneous intracerebral hemorrhage.
Donor spontaneous intracerebral hemorrhage (ICH) is a potential risk factor for morbidity and mortality after cardiac transplantation. We hypothesized that donor ICH is associated with systemic up-regulation of angiotensin II receptor type 1 (AT1R). We evaluated mRNA expression of AT1R and AT2R in donor spleen lymphocytes and in heart biopsies from 20 recipients of hearts from donors with spontaneous ICH which were compared with 20 recipients from trauma donors. Heart biopsies showed 4.7-fold increased mRNA expression of AT1R (p < 0.0001) in the ICH group compared with the Trauma group. The ICH group also showed 2.6-fold (p < 0.01) increased mRNA expression of AT1R in the donor spleen lymphocytes, suggesting the presence of systemic activation before transplantation. At 1 year, the ICH group had increased coronary vasculopathy by vascular ultrasound. Using multivariate regression analysis, mRNA expression of AT1R in the donor spleen lymphocytes was found to be a strong independent predictor of transplant vasculopathy (odds ratio = 4.397, CI = 1.243-15.553, adjusted p = 0.02). This is the first report to describe splenic up-regulation of AT1R in the presence of spontaneous ICH and its association with subsequent development of transplant vasculopathy.


The effect of LNA nucleobases as enhancers for the binding of amiloride to an abasic site in DNA/DNA and DNA/RNA duplexes.
We report on a significant effect of locked nucleic acid (LNA) nucleobases on the binding of amiloride for abasic site (AP)-containing DNA duplexes. Fluorescence titration experiments showed that the binding affinity of amiloride for the target thymine (T) opposite an AP site significantly improves for the DNA duplexes possessing LNA nucleobases that flank the AP site, compared to the corresponding normal DNA duplexes. In particular, LNA flanking nucleobases on both 5'- and 3'-sides of the AP site are found to be effective for the enhancement of the binding affinity. From thermodynamic characterization of the amiloride binding, the loss in the binding entropy is remarkably reduced for the LNA-containing DNA duplexes, which is indeed responsible for the enhanced affinity of amiloride. Moreover, such an effect of LNA nucleobases was also observed for amiloride binding to DNA/RNA hybrid duplexes.


Abnormal liver development and resistance to 2,3,7,8-tetrachlorodibenzo-p-dioxin toxicity in mice carrying a mutation in the DNA-binding domain of the aryl hydrocarbon receptor.
The aryl hydrocarbon receptor (AHR) is known for its role in the adaptive and toxic responses to a large number of environmental contaminants, as well as its role in hepatovascular development. The classical AHR pathway involves ligand binding, nuclear translocation, heterodimerization with the AHR nuclear translocator (ARNT), and binding of the heterodimer to dioxin response elements (DREs), thereby modulating the transcription of an array of genes. The AHR has also been implicated in signaling events independent of nuclear localization and DNA binding, and it has been suggested that such pathways may play important roles in the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Here, we report the generation of a mouse model that expresses an AHR protein capable of ligand binding, interactions with chaperone proteins, functional heterodimerization with ARNT, and nuclear translocation, but is unable to bind DREs. Using this model, we provide evidence that DNA binding is required AHR-mediated liver development, as Ahr(dbd/dbd) mice exhibit a patent ductus venosus, similar to what is seen in Ahr(-/-) mice. Furthermore, Ahr(dbd/dbd) mice are resistant to TCDD-induced toxicity for all endpoints tested. These data suggest that DNA binding is necessary for AHR-mediated developmental and toxic signaling.


p21WAF1 expression by an activator of protein kinase C is regulated mainly at the post-transcriptional level in cells lacking p53: important role of RNA stabilization.
p21(WAF1) inhibits cyclin-cyclin-dependent kinase (Cdk) complexes, causing cell cycle arrest. p21(WAF1) contains p53-binding sites in its promoter and expression of p21(WAF1) is induced by functional p53. In the present work, we have studied the role of protein kinase C (PKC) in the induction of p21(WAF1) and show that induction of p21(WAF1) expression can occur by activation of PKC in cells having no p53. Human ovarian carcinoma cells, SKOV-3, lack p53 protein and PMA, a potent activator of PKC, did not induce p53. PMA increased the expression of p21(WAF1) mRNA both in these cells and in other cells which do not contain p53 (THP-1 and U937). Treatment of human embryonic fibroblasts, WI38, with PMA also induced the accumulation of p21(WAF1) without affecting p53 levels. However, PMA did not increase levels of p21(WAF1) mRNA in cells where either the PKC or the mitogen-activated protein kinase pathway was blocked. Furthermore, treatment of cells with various phorbol ester derivatives which activate PKC resulted in the induction of p21(WAF1) in SKOV-3 cells. In contrast, phorbol esters which do not activate PKC failed to induce p21(WAF1) expression. PMA increased the transcriptional rate of p21(WAF1) and activated the transcription of a luciferase reporter gene, controlled by the p21 promoter, in SKOV-3 cells with or without a p53 consensus-binding sequence. By contrast, PMA markedly stabilized p21(WAF1) mRNA; the half-life (t1/2) of p21(WAF1) in PMA-treated cells was >8 h compared with <1 h in untreated cells. These findings provide evidence that the PKC pathway induces expression of p21(WAF1) independently of p53. Our present study also suggests that the accumulation of p21(WAF1) transcripts by PMA occurs mainly at post-transcriptional level.


The process of developing a community-based research agenda with lesbian, gay, bisexual, transgender and queer youth in the Northwest Territories, Canada.
Youth in Canada's Northwest Territories (NWT) experience sexual and mental health disparities. Higher rates of sexual and mental health concerns among lesbian, gay, bisexual, transgender and queer (LGBTQ) youth in comparison with heterosexual and cisgender peers have been associated with stigma and discrimination. Although LGBTQ youth in the NWT are situated at the nexus of Northern and LGBTQ health disparities, there is little known about their health, well-being and experiences of stigma. This short communication discusses the process of developing a LGBTQ youth community-based research programme in the NWT. We developed an interdisciplinary research team of LGBTQ and allied young adults, including indigenous and non-indigenous researchers, community organisers and service providers in the NWT. We conducted meetings in Yellowknife with LGBTQ youth (n=12) and key stakeholders (n=15), including faculty, students, community groups and health and social service providers. Both meetings included LGBTQ and allied participants who were LGBTQ, indigenous, youth and persons at the intersection of these identities. LGBTQ youth participants discussed community norms that devalued same sex identities and stigma surrounding LGBTQ-specific services and agencies. Stigma among LGBT youth was exacerbated for youth in secondary schools, gender non-conforming and transgender youth and young gay men. In the stakeholder meeting, service providers discussed the importance of integrating LGBTQ issues in youth programmes, and LGBTQ community groups expressed the need for flexibility in service delivery to LGBTQ youth. Stakeholders identified the need to better understand the needs of indigenous LGBTQ youth in the NWT. Community-based LGBTQ groups, researchers and health and social service providers are interested in addressing LGBTQ youth issues in the NWT. The emergence of LGBTQ community building, support groups and activism in Northern Canada suggests that this is an opportune time to explore LGBTQ youth health.


Anatomic variations and technical problems of the anterolateral thigh flap: a report of 74 cases.
We have transferred 74 free or pedicled anterolateral thigh flaps, including those combined with other flaps, for reconstruction of various types of defects. We report several anatomic variations of the lateral circumflex arterial system and discuss some technical problems with this flap. Septocutaneous perforators were found in 28 of 74 cases (37.8 percent), and no perforators were found in 4 cases (5.4 percent). In the 70 cases with perforators, 171 tiny cutaneous perforators (an average of 2.31 per case) were found. Musculocutaneous perforators (81.9 percent) were much more common than septocutaneous perforators (18.1 percent). Perforators were concentrated near the midpoint of the lateral thigh, and the selection of perforators as nutrient vessels for the anterolateral thigh flap was related to the length of the pedicle and the thickness of the skin flap. Anatomic variations of the branching pattern of perforators were classified into eight types. Flaps with perforators that arise directly from the profunda femoris artery are difficult to combine with other free flaps. Because the perforators are extremely small and tend to thrombose soon after congestion develops, these flaps are difficult to salvage with recirculation surgery. Therefore, several perforators should be included with the flap, if possible. The descending artery of the lateral circumflex femoral artery was always accompanied by two veins with different back-flow strengths. Therefore, veins for microsurgical anastomosis must be chosen carefully. Because it is nourished by several perforators arising from the descending artery, the vastus lateralis muscle can be combined with the anterolateral thigh flap. However, splitting the muscle longitudinally without harvesting its blood supply is complicated because its fibers are oblique. The rectus femoris muscle can also be combined with the anterolateral thigh flap, but its pedicle is short and its origin is very near the site of anastomosis. When the anterolateral thigh flap is combined with the tensor fasciae latae musculocutaneous flap, the large skin area of the lateral part of thigh can be transferred to repair the massive defects. The anterolateral thigh flap has many advantages and can be used to reconstruct many types of defect. However, anatomic variations must be considered if the flap is to be used safely and reliably.


Does the Addition of Salubrinal to in vitro Maturation Medium Enhance Bovine Blastocyst Yields and Embryo Cryotolerance?
Modifications to in vitro maturation (IVM) media are made to improve rates of blastocyst formation and quality of mammalian embryos. Embryo quality is an important factor in the viability of embryos following cryopreservation. Salubrinal is a specific inhibitor of endoplasmic reticulum stress-induced apoptosis in eukaryotic cells. Here, it was added to IVM medium in an attempt to increase blastocyst formation and to enhance embryo quality in cattle. To assess the effect on blastocyst formation and cryotolerance of the supplementation of salubrinal to bovine IVM medium. Cumulus-oocyte complexes (COCs) collected from slaughterhouse ovaries were assigned randomly to two groups, either cultured in IVM medium that was supplemented with 400 mM salubrinal (treated, 262 COCs) or that was not supplemented (control, 263 COCs). All oocytes of the matured COCs were fertilized in vitro with sperm from the same proven bull and cultured for 6-7 d. At the time of blastocyst collection, expanded blastocysts were chosen for cryopreservation, while early, hatching and hatched blastocysts and those of poor quality were not used. There were 83 expanded blastocysts classified to be of good quality in both the control and salubrinal-treated groups that were subjected to vitrification. After 5 to 10 months of cold storage, the embryos were warmed and cultured in vitro for 24 h to assess the survival rate and for 48 h to assess the hatching rate. The blastocyst developmental rate in the salubrinal-treated group was similar to that in the control group, 61.5% compared with 62.7% (P > 0.05). The survival rate of blastocysts after vitrification was also similar, at or very close to 100%. In addition, there was no statistically significant difference in the hatching rate of expanded blastocysts derived from the COCs cultured with (treated) and without (control) addition of salubrinal to the IVM medium (91.6% compared with 85.5%; P > 0.05). Supplementation of salubrinal to the IVM medium neither improved nor reduced rates of bovine blastocyst formation and of embryo cryotolerance as determined by post-warming viability.


Identification of an outer membrane protein involved in utilization of hemoglobin-haptoglobin complexes by nontypeable Haemophilus influenzae.
A recombinant plasmid containing a 6.5-kb fragment of nontypeable Haemophilus influenzae (NTHI) chromosomal DNA was shown to confer a hemoglobin-haptoglobin-binding phenotype on Escherichia coli. Use of a mini-Tn10kan transposon for random insertion mutagenesis of this recombinant plasmid allowed localization of the NTHI DNA responsible for this hemoglobin-haptoglobin-binding phenotype to a 3.5-kb PstI-XhoI fragment within the 6.5-kb NTHI DNA insert. When this mutagenized NTHI DNA fragment was used to transform the wild-type NTHI strain, the resultant kanamycin-resistant mutant exhibited significantly decreased abilities to bind hemoglobin-haptoglobin and utilize it as a source of heme for aerobic growth in vitro. This mutant also lacked expression of a 115-kDa outer membrane protein that was present in the wild-type parent strain. Transformation of this mutant with wild-type NTHI chromosomal DNA restored the abilities to bind and utilize hemoglobin-haptoglobin and to express the 115-kDa outer membrane protein. Nucleotide sequence analysis of the relevant NTHI DNA revealed the presence of a gene, designated hhuA, that encoded a predicted 117,145-Da protein. The HhuA protein exhibited features typical of a TonB-dependent outer membrane receptor and had significant identity with the hemoglobin receptors of both Haemophilus ducreyi and Neisseria meningitidis.


Cerebellar stem cells act as medulloblastoma-initiating cells in a mouse model and a neural stem cell signature characterizes a subset of human medulloblastomas.
Cells with stem cell properties have been isolated from various areas of the postnatal mammalian brain, most recently from the postnatal mouse cerebellum. We show here that inactivation of the tumor suppressor genes Rb and p53 in these endogenous neural stem cells induced deregulated proliferation and resistance to apoptosis in vitro. Moreover, injection of these cells into mice formed medulloblastomas. Medulloblastomas are the most common malignant brain tumors of childhood, and despite recent advances in treatment they are associated with high morbidity and mortality. They are highly heterogeneous tumors characterized by a diverse genetic make-up and expression profile as well as variable prognosis. Here, we describe a novel ontogenetic pathway of medulloblastoma that significantly contributes to understanding their heterogeneity. Experimental medulloblastomas originating from neural stem cells preferentially expressed stem cell markers Nestin, Sox2 and Sox9, which were not expressed in medulloblastomas originating from granule-cell-restricted progenitors. Furthermore, the expression of these markers identified a subset of human medulloblastomas associated with a poorer clinical outcome.


Prepackaged therapy for urethritis: the "MSTOP" experience in Cameroon.
The social marketing of STD treatment may be a strategy to increase the availability of effective therapy for urethritis in male patients. To evaluate a pilot project of social marketing of urethritis treatment packages. The project, initially designed for over the counter sale in private pharmacies, was finally restricted by national health authorities to primary healthcare settings in Yaoundé and Douala, Cameroon. Monthly sales of packages containing antibiotics, condoms, partner referral cards, and written information on STDs were monitored by the social marketing agency. Structured interviews were conducted with a sample of traceable patients who had consulted for urethritis. Structured interviews completed by focus group discussions were conducted among healthcare providers. Interview findings were further validated by a "mystery patient" survey, using surrogate patients. Lastly, 15 key informants among the decision markers involved in the project were interviewed in depth. Local independent consultants carried out the whole evaluation. A total of 1392 treatment packages were sold in 10 months. Patients who had purchased the package reported high compliance with the treatment, with 99% taking the single dose of cefuroxime-axetil and 83% completing the course of doxycycline. 76% notified all or some partners, and 84% of those who had sex during treatment used condoms. In contrast, only 27% of trained healthcare providers prescribed "MSTOP". They questioned the omission of laboratory diagnosis, the selection of antibiotics, and the duration of therapy. Public health authorities were also sceptical about the choice of antibiotics and viewed the initial project as an overt encouragement of self medication. Although the MSTOP project was not implemented in the way it had initially been designed, it highlighted the patients' interest in the product. Public health authorities in Cameroon should have been made aware of the limitations of the formal sector's response to STD care among men before over the counter sale of prepackaged therapy could have been considered as an alternative approach to inadequate self medication.


Reliability of interacting quantum atoms (IQA) data computed from post-HF densities: impact of the approximation used.
The performance of BBC1, BBC2 and Müller approximations, in terms of reliability of IQA data, was investigated at the CCSD, CCSD(T) and MP2 levels using glycol, as a case study, in interpreting the relative stability of its conformers, one with H-bond type intramolecular interaction and the other with a steric clash between two O-atoms. The CCSD/BBC1 level appeared to be perfectly suited as a reference needed to evaluate all possible levels of theory/approximation combinations (LoT/LoA). We found the reliability trend LoT/BBC1 > LoT/BBC2 > LoT/Müller (as well as its origin) and concluded that the Müller approximation should not be used when the accuracy of IQA-defined energy terms is considered. Moreover, we have established that the requirement of reproducing, by IQA calculations, electronic energy is desirable but not a necessary requirement when a comparative approach is used, such as in FAMSEC-based analysis (FAMSEC = fragment attributed molecular system energy change). A new criterion is proposed to assess the quality of IQA data for comparative analyses, ΔE(IQA) ≈ ΔE, where ΔE(IQA) and ΔE are the IQA and electronic energy differences, respectively, between the fin-state and ref-state of a molecular system. The closer ΔE(IQA) approaches ΔE, the closer the FAMSEC data approach values obtained at the exceptionally well performing CCSD/BBC1 level, regardless of the LoT/LoA combination used. Importantly, the MP2/BBC1 level performed nearly as well as the CCSD/BBC1 level in comparative studies. The origin of the MP2/BBC1 approximation's exceptional and the MP2/Müller approximations's acceptable performance in explaining the relative stability of glycol conformers has been uncovered and discussed in detail.


Efficient complexation between pillar[5]arenes and neutral guests: from host-guest chemistry to functional materials.
Since their discovery in 2008, pillar[n]arenes have been a popular family of macrocyclic arene hosts due to their accessible one-step synthesis, convenient functionalization, symmetrical prism structures and perfect cavity host-guest properties. Compared with other macrocyclic hosts, the most peculiar recognition behavior of pillararenes is the strong binding affinities of pillar[5]arenes (P5As) towards neutral guests in organic media, which is unfeasible for classic crown ethers and calixarenes. The intriguing properties have found extensive applications in many fields from supramolecular chemistry to materials science. This feature article provides a detailed summary of the molecular recognition of P5As and neutral guests, where the driving forces, binding mechanisms, and binding selectivities are comprehensively discussed. Furthermore, brief highlights of research progress in the functional applications based on the neutral guest⊂P5A motifs were also discussed, including the construction of complex topological superstructures (e.g. rotaxanes, catenanes and daisy chains), supramolecular polymers, and functional materials.


The effect of a non-talking rule on the sound level and perception of patients in an outpatient infusion center.
Noise is a common problem in hospitals, and it is known that social behavior can influence sound levels. The aim of this naturally-occurring field experiment was to assess the influence of a non-talking rule on the actual sound level and perception of patients in an outpatient infusion center. In a quasi-randomized trial two conditions were compared in real life. In the control condition, patients (n = 137) were allowed to talk to fellow patients and visitors during the treatment. In the intervention condition patients (n = 126) were requested not to talk to fellow patients and visitors during their treatment. This study measured the actual sound levels in dB(A) as well as patients' preferences regarding sound and their perceptions of the physical environment, anxiety, and quality of health care. A linear-mixed-model showed a statistically significant, but rather small reduction of the non-talking rule on the actual sound level with an average of 1.1 dB(A). Half of the patients preferred a talking condition (57%), around one-third of the patients had no preference (36%), and 7% of the patients preferred a non-talking condition. Our results suggest that patients who preferred non-talking, perceived the environment more negatively compared to the majority of patients and perceived higher levels of anxiety. Results showed no significant effect of the experimental conditions on patient perceptions. In conclusion, a non-talking rule of conduct only minimally reduced the actual sound level and did not influence the perception of patients.


Occupational exposure to benzene at the ExxonMobil refinery in Baytown, TX (1978-2006).
Although occupational benzene exposure of refinery workers has been studied for decades, no extensive analysis of historical industrial hygiene data has been performed focusing on airborne concentrations at specific refineries and tasks. This study characterizes benzene exposures at the ExxonMobil Baytown, TX, refinery from 1978 to 2006 to understand the variability in workers' exposures over time and during different job tasks. Exposures were grouped by operational status, job title, and tasks. More than 9000 industrial hygiene air samples were evaluated; approximately 4000 non-task (> 3 h) and 1000 task-related (< 3 h) personal samples were considered. Each sample was assigned to one of 27 job titles, 29 work areas, and 16 task bins (when applicable). Process technicians were sampled most frequently, resulting in the following mean benzene concentrations by area: hydrofiner (n=245, mean=1.3 p.p.m.), oil movements (n=286, mean=0.23 p.p.m.), reformer (n=575, mean=0.10 p.p.m.), tank farm (n=9, mean=0.65 p.p.m.), waste treatment (n=446, mean=0.13 p.p.m.), and other areas (n=460, mean=0.062 p.p.m.). The most frequently sampled task was sample collection (n=218, mean=0.40 p.p.m.). Job title and area did not significantly impact task-related exposures. Airborne concentrations were significantly lower after 1990 than before 1990. Results of this task-focused study may be useful when analyzing benzene exposures at other refineries.


Extremely bulky copper(i) complexes of [HB(3,5-{1-naphthyl}2pz)3]- and [HB(3,5-{2-naphthyl}2pz)3]- and their self-assembly on graphene.
The synthesis and characterization, using NMR (1H and 13C), infrared spectroscopy, and X-ray crystallography, of the ethene and carbon monoxide copper(i) complexes of hydridotris(3,5-diphenylpyrazol-1-yl)borate ([TpPh2]-) and the two new ligands hydridotris(3,5-bis(1-naphthyl)pyrazol-1-yl)borate ([Tp(1Nt)2]-) and hydridotris(3,5-bis-(2-naphthyl)pyrazol-1-yl)borate ([Tp(2Nt)2]-) are described. X-ray crystal structures are presented of [Cu(TpPh2)(C2H4)] and [Cu(Tp(2Nt)2)(C2H4)]. The compound [Cu(TpPh2)(C2H4)] features interactions between the protons of the ethene ligand and the π-electron clouds of the phenyl substituents that make up the binding pocket surrounding the copper(i) center. These dipolar interactions result in strongly upfield shifted signals of the ethene protons in 1H-NMR. [Cu(Tp(1Nt)2)(CO)] and [Cu(Tp(2Nt)2)(CO)] were examined using infrared spectroscopy and were found to have CO stretching vibrations at 2076 and 2080 cm-1 respectively. The copper(i) carbonyl complexes form self-assembled monolayers when drop cast onto HOPG and thin multilayers of a few nanometers thickness when dip coated onto graphene. General macroscopic trends such as the different tendencies to crystallize observed in the complexes of the two naphthyl-substituted ligands appear to extend well to the nanoscale where a well-organized monolayer could be observed of [Cu(Tp(2Nt)2)(CO)].


Placental PAGs: gene origins, expression patterns, and use as markers of pregnancy.
Pregnancy-associated glycoproteins (PAGs) are abundantly expressed products of the placenta of species within the Cetartiodactyla order (even-toed ungulates). They are restricted to this order and they are particularly numerous in the Bovidae. The PAGs exhibit a range of temporal and spatial expression patterns by the placental trophoblasts and probably represent a group of related proteins that perform a range of distinct functions in the epitheliochorial and synepitheliochorial placental forms. This review presents an overview of the origins of the PAGs, a summary of PAG expression patterns, and their use as markers of pregnancy status. Speculations about their putative role(s) in pregnancy are also presented.


Enantioselective synthesis of gem-diarylalkanes by transition metal-catalyzed asymmetric arylations (TMCAAr).
Chiral gem(1,1)-diaryl containing tertiary or quaternary stereogenic centers are present in many natural products and important pharmacophores. While numerous catalytic asymmetric methods enable access to 1,1-diaryl motifs, transition metal-catalyzed asymmetric arylations (TMCAAr) are one of the most powerful methods to prepare enantiopure gem-diarylalkane compounds. The main methodology includes enantioselective 1,2- or 1,4-additions across C[double bond, length as m-dash]O, C[double bond, length as m-dash]N and C[double bond, length as m-dash]C bonds by arylmetallic reagents; aryl cross-couplings of olefins, benzylic (pseudo)halides and aziridines; asymmetric aryl substitution reactions of allylic substrates; and isotopic benzylic C-H arylation.


Neural Circuitry of Impaired Emotion Regulation in Substance Use Disorders.
Impaired emotion regulation contributes to the development and severity of substance use disorders (substance disorders). This review summarizes the literature on alterations in emotion regulation neural circuitry in substance disorders, particularly in relation to disorders of negative affect (without substance disorder), and it presents promising areas of future research. Emotion regulation paradigms during functional magnetic resonance imaging are conceptualized into four dimensions: affect intensity and reactivity, affective modulation, cognitive modulation, and behavioral control. The neural circuitry associated with impaired emotion regulation is compared in individuals with and without substance disorders, with a focus on amygdala, insula, and prefrontal cortex activation and their functional and structural connectivity. Hypoactivation of the rostral anterior cingulate cortex/ventromedial prefrontal cortex (rACC/vmPFC) is the most consistent finding across studies, dimensions, and clinical populations (individuals with and without substance disorders). The same pattern is evident for regions in the cognitive control network (anterior cingulate and dorsal and ventrolateral prefrontal cortices) during cognitive modulation and behavioral control. These congruent findings are possibly related to attenuated functional and/or structural connectivity between the amygdala and insula and between the rACC/vmPFC and cognitive control network. Although increased amygdala and insula activation is associated with impaired emotion regulation in individuals without substance disorders, it is not consistently observed in substance disorders. Emotion regulation disturbances in substance disorders may therefore stem from impairments in prefrontal functioning, rather than excessive reactivity to emotional stimuli. Treatments for emotion regulation in individuals without substance disorders that normalize prefrontal functioning may offer greater efficacy for substance disorders than treatments that dampen reactivity.


Analysis of the effects of bias phase and wavelength choice on the design of dual-wavelength diffractive optical elements.
Diffractive optical elements (DOEs) are often used in pattern formation for display purposes. Constructing these images from two or more colors greatly enhances their visual effect. To achieve this with DOEs is not simple, as they are inherently wavelength specific. We discuss an algorithm for designing quantized elements that produce distinct intensity patterns in the far field for two wavelengths. The benefits of applying bias phase to the dual-wavelength problem are investigated. The difference between the best and the worst choice of bias phase is shown to produce a variation of up to 2% in the efficiency. The mean square error can vary by up to a factor of 2 between the best and the worst case. It is also critically important to understand how the values of the two wavelengths affect the result. We present an analysis of how choosing different pairs of wavelengths in the design process affects the quality of our results.


HIV-1 RNAs are Not Part of the Argonaute 2 Associated RNA Interference Pathway in Macrophages.
MiRNAs and other small noncoding RNAs (sncRNAs) are key players in post-transcriptional gene regulation. HIV-1 derived small noncoding RNAs (sncRNAs) have been described in HIV-1 infected cells, but their biological functions still remain to be elucidated. Here, we approached the question whether viral sncRNAs may play a role in the RNA interference (RNAi) pathway or whether viral mRNAs are targeted by cellular miRNAs in human monocyte derived macrophages (MDM). The incorporation of viral sncRNAs and/or their target RNAs into RNA-induced silencing complex was investigated using photoactivatable ribonucleoside-induced cross-linking and immunoprecipitation (PAR-CLIP) as well as high-throughput sequencing of RNA isolated by cross-linking immunoprecipitation (HITS-CLIP), which capture Argonaute2-bound miRNAs and their target RNAs. HIV-1 infected monocyte-derived macrophages (MDM) were chosen as target cells, as they have previously been shown to express HIV-1 sncRNAs. In addition, we applied small RNA deep sequencing to study differential cellular miRNA expression in HIV-1 infected versus non-infected MDMs. PAR-CLIP and HITS-CLIP data demonstrated the absence of HIV-1 RNAs in Ago2-RISC, although the presence of a multitude of HIV-1 sncRNAs in HIV-1 infected MDMs was confirmed by small RNA sequencing. Small RNA sequencing revealed that 1.4% of all sncRNAs were of HIV-1 origin. However, neither HIV-1 derived sncRNAs nor putative HIV-1 target sequences incorporated into Ago2-RISC were identified suggesting that HIV-1 sncRNAs are not involved in the canonical RNAi pathway nor is HIV-1 targeted by this pathway in HIV-1 infected macrophages.


CD32+ and PD-1+ Lymph Node CD4 T Cells Support Persistent HIV-1 Transcription in Treated Aviremic Individuals.
A recent study conducted in blood has proposed CD32 as the marker identifying the "elusive" HIV reservoir. We have investigated the distribution of CD32+ CD4 T cells in blood and lymph nodes (LNs) of HIV-1-uninfected subjects and viremic untreated and long-term-treated HIV-1-infected individuals and their relationship with PD-1+ CD4 T cells. The frequency of CD32+ CD4 T cells was increased in viremic compared to treated individuals in LNs, and a large proportion (up to 50%) of CD32+ cells coexpressed PD-1 and were enriched within T follicular helper (Tfh) cells. We next investigated the role of LN CD32+ CD4 T cells in the HIV reservoir. Total HIV DNA was enriched in CD32+ and PD-1+ CD4 T cells compared to CD32- and PD-1- cells in both viremic and treated individuals, but there was no difference between CD32+ and PD-1+ cells. There was no enrichment of latently infected cells with inducible HIV-1 in CD32+ versus PD-1+ cells in antiretroviral therapy (ART)-treated individuals. HIV-1 transcription was then analyzed in LN memory CD4 T cell populations sorted on the basis of CD32 and PD-1 expression. CD32+ PD-1+ CD4 T cells were significantly enriched in cell-associated HIV RNA compared to CD32- PD-1- (averages of 5.2-fold in treated individuals and 86.6-fold in viremics), CD32+ PD-1- (2.2-fold in treated individuals and 4.3-fold in viremics), and CD32- PD-1+ (2.2-fold in ART-treated individuals and 4.6-fold in viremics) cell populations. Similar levels of HIV-1 transcription were found in CD32+ PD-1- and CD32- PD-1+ CD4 T cells. Interestingly, the proportion of CD32+ and PD-1+ CD4 T cells negatively correlated with CD4 T cell counts and length of therapy. Therefore, the expression of CD32 identifies, independently of PD-1, a CD4 T cell population with persistent HIV-1 transcription and coexpression of CD32 and PD-1, the CD4 T cell population with the highest levels of HIV-1 transcription in both viremic and treated individuals.IMPORTANCE The existence of long-lived latently infected resting memory CD4 T cells represents a major obstacle to the eradication of HIV infection. Identifying cell markers defining latently infected cells containing replication-competent virus is important in order to determine the mechanisms of HIV persistence and to develop novel therapeutic strategies to cure HIV infection. We provide evidence that PD-1 and CD32 may have a complementary role in better defining CD4 T cell populations infected with HIV-1. Furthermore, CD4 T cells coexpressing CD32 and PD-1 identify a CD4 T cell population with high levels of persistent HIV-1 transcription.


Utility and Significance of Gadolinium-Based Contrast Enhancement in Posterior Reversible Encephalopathy Syndrome.
Posterior reversible encephalopathy syndrome is a clinicoradiologic syndrome. Literature regarding associated factors and the prognostic significance of contrast enhancement in posterior reversible encephalopathy syndrome is sparse. This study set out to evaluate an association between the presence of enhancement in posterior reversible encephalopathy syndrome and various clinical factors in a large series of patients with this syndrome. From an MR imaging report search that yielded 176 patients with clinically confirmed posterior reversible encephalopathy syndrome between 1997 and 2014, we identified 135 patients who had received gadolinium-based contrast. The presenting symptoms, etiology, clinical follow-up, and maximum systolic and diastolic blood pressures within 1 day of MR imaging were recorded. MRIs were reviewed for parenchymal hemorrhage, MR imaging severity, and the presence and pattern of contrast enhancement. Statistical analyses evaluated a correlation between any clinical features and the presence or pattern of enhancement. Of 135 included patients (67.4% females; age range, 7-82 years), 59 (43.7%) had contrast enhancement on T1-weighted MR imaging, the most common pattern being leptomeningeal (n = 24, 17.8%) or leptomeningeal plus cortical (n = 21, 15.6%). Clinical outcomes were available in 96 patients. No significant association was found between the presence or pattern of enhancement and any of the variables, including sex, age, symptom, MR imaging severity, blood pressure, or outcome (all P > .05 after Bonferroni correction). The presence or pattern of enhancement in posterior reversible encephalopathy syndrome is not associated with any of the tested variables. However, an association was found between MR imaging severity and clinical outcome.


The catechol-O-methyltransferase polymorphism: relations to the tonic-phasic dopamine hypothesis and neuropsychiatric phenotypes.
Diverse phenotypic associations with the catechol-O-methyltransferase (COMT) Val158Met polymorphism have been reported. We suggest that some of the complex effects of this polymorphism be understood from the perspective of the tonic-phasic dopamine (DA) hypothesis. We hypothesize that the COMT Met allele (associated with low enzyme activity) results in increased levels of tonic DA and reciprocal reductions in phasic DA in subcortical regions and increased D1 transmission cortically. This pattern of effects is hypothesized to yield increased stability but decreased flexibility of neural network activation states that underlie important aspects of working memory and executive functions; these effects may be beneficial or detrimental depending on the phenotype, a range of endogenous factors, and environmental exigencies. The literature on phenotypic associations of the COMT Val158Met polymorphism is reviewed, highlighting areas where this hypothesis may have explanatory value, and pointing to possible directions for refinement of relevant phenotypes and experimental evaluation of this hypothesis.