Widespread brain tau and its association with ageing, Braak stage and Alzheimer's dementia.
See Herholz (doi:10.1093/brain/awx340) for a scientific commentary on this article.Autopsy data have proposed that a topographical pattern of tauopathy occurs in the brain with the development of dementia due to Alzheimer's disease. We evaluated the findings of tau-PET to better understand neurofibrillary tangle development as it is seen in cognitively unimpaired and impaired individuals. The evolution of Alzheimer's disease tauopathy in cognitively unimpaired individuals needs to be examined to better understand disease pathogenesis. Tau-PET was performed in 86 cognitively impaired individuals who all had abnormal amyloid levels and 601 cognitively unimpaired individuals. Tau-PET findings were assessed for relationships with clinical diagnosis, age, and regional uptake patterns relative to Braak stage. Regional and voxel-wise analyses were performed. Topographical findings from tau-PET were characterized using hierarchical clustering and clinical characteristic-based subcategorization. In older cognitively unimpaired individuals (≥50 years), widespread, age-related elevated tau signal was seen among those with normal or abnormal amyloid status as compared to younger cognitively unimpaired individuals (30-49 years). More frequent regional tau signal elevation throughout the brain was seen in cognitively unimpaired individuals with abnormal versus normal amyloid. Elevated tau signal was seen in regions that are considered high Braak Stage in cognitively unimpaired and cognitively impaired individuals. Hierarchical clustering and clinical characteristic-based categorizations both showed different patterns of tau signal between groups such as greater tau signal in frontal regions in younger onset Alzheimer's disease dementia participants (most of whom had a dysexecutive clinical presentation). Tau-PET signal increases modestly with age throughout the brain in cognitively unimpaired individuals and elevated tau is seen more often when amyloid brain accumulation is present. Tau signal patterns in cognitively unimpaired correspond to early Braak stage but also suggest tangle involvement in extra-medial temporal and extra-temporal regions that are considered more advanced in the Braak scheme even when amyloid negative. Our findings also suggest the possibility of widespread development of early tangle pathology rather than a pattern defined exclusively by adjacent, region-to-region spread, prior to onset of clinical symptoms. Distinct patterns of neurofibrillary tangle deposition in younger-onset Alzheimer's disease dementia versus older-onset Alzheimer's disease dementia provide evidence for variability in regional tangle deposition patterns and demonstrate that different disease phenotypes have different patterns of tauopathy. Pathological correlation with imaging is needed to assess the implications of these observations.


Identification and characterization of novel promoters in the genome of human papillomavirus type 18.
Most studies on the regulation of gene expression in human papillomaviruses (HPV) have focused on the promoter for the early genes E6 and E7. This promoter is located at the junction between the long control region and the E6 open reading frame. RNA mapping studies have suggested that additional promoters may exist in other parts of the genome. In this study, we used a combination of transcription in vitro and an analysis of RNA produced in vivo in transfected cells to identify three novel promoters in the genome of human papillomavirus type 18. These promoters are located in front of the E2 gene (P2598), within the E2 coding sequences (P3036), and at the end of the L2 open reading frame (P5600). They were active in HeLa cells, as shown by a chloramphenicol acetyltransferase assay. The activity of the P3036 promoter was stimulated by the bovine papillomavirus type 1 E2 protein.


Localization of atrial natriuretic peptide in caveolae of in situ atrial myocytes.
The plasma membrane-associated non-clathrin-coated vesicles called caveolae are multifunctional organelles thought to be implicated in the sequestration and transport of small molecules (potocytosis) as well as in the binding of Ca2+ ions, signal transduction, and processing of hormonal and mechanosensitive signals. We have previously suggested that the apparent contiguity of caveolar and atrial granule membranes observed in electron micrographs of in situ mouse atrial myocytes might reflect externalization of atrial natriuretic peptide through caveolae. Using Tokuyasu's classic technique, we now show by immunoelectron microscopy of glutaraldehyde-fixed and cryosectioned mouse and rat atria that antibody against atrial natriuretic peptide prohormone is present within caveolae of in situ atrial myocytes. We confirm this intracaveolar localization by stereoimaging colloidal gold-labeled antibody to the prohormone in electron micrographs of glutaraldehyde/osmium tetroxide-fixed positively stained atrial thin sections. Because profiles of caveolae were rarely immunolabeled with antibody against atrial peptide unless there was a profile of an immunolabeled atrial granule nearby in the subjacent cytoplasm, we concluded that the intracaveolar hormone was derived predominantly from a direct interaction of atrial granules with caveolae. Perturbations that markedly increase the rate of natriuretic peptide secretion via the regulated pathway, including atrial stretch, contractions, and increased external Ca2+ concentration, failed to alter caveolar immunostaining. These results suggest that atrial peptide can pass from atrial granules into caveolae by transiently open pathways between the interiors of granules and caveolae. The results are interpreted as suggesting the presence of a second pathway for externalization of atrial natriuretic peptide through caveolae in addition to the classic pathway for regulated atrial peptide secretion at noncaveolar plasmalemma.


Two distinct EF-Tu epitopes induce immune responses in rice and Arabidopsis.
Plants sense potential pathogens by recognizing conserved pathogen-associated molecular patterns (PAMPs) that cause PAMP-triggered immunity (PTI). We previously reported that rice recognizes flagellin from the rice-incompatible N1141 strain of Acidovorax avenae and subsequently induces immune responses. Cell extracts isolated from flagellin-deficient N1141 (Δfla1141) still induced PTI responses, suggesting that Δfla1141 possesses an additional PAMP distinct from flagellin. Here, we show that elongation factor Tu (EF-Tu), one of the most abundant bacterial proteins, acts as a PAMP in rice and causes several PTI responses. In Brassicaceae species, EF-Tu and an N-acetylated peptide comprising the first 18 amino acids of the N-terminus, termed elf18, are fully active as inducers of PTI responses. By contrast, elf18 did not cause any immune responses in rice, whereas an EF-Tu middle region comprising Lys176 to Gly225, termed EFa50, is fully active as a PAMP in rice. In the leaves of rice plants, EF-Tu induced H2O2 generation and callose deposition, and also triggered resistance to coinfection with pathogenic bacteria. Taken together, these data demonstrate that rice recognizes EFa50, which is distinct from elf18, and that this epitope induces PTI responses.


Cortical Fenestration for Megaprosthesis Stem Revision.
The most common modes of failure for megaprostheses are aseptic loosening followed by periprosthetic infection and stem fracture. Surgical technique for bone and implant exposure is controversial and may influence the success of revision and the need for additional future revisions. The purpose of this study was to evaluate the effectiveness of cortical fenestration for megaprosthesis revision, particularly for stem fracture. From 1985-2014, 196 adult and pediatric patients underwent limb salvage with a distal femoral or proximal tibial megaprosthesis (109 cemented, 87 pressfit). A retrospective chart review was performed to assess the rate of revision based on cemented or pressfit fixation and the use of a cortical window to extract the failed stem. Results: 27% (29 of 109) of cemented and 18% (16 of 87) of pressfit implants were revised for stem failure. The reasons for revision in the cemented group were loosening (62%), infection (24%), and stem fracture (13%). In the pressfit group, the reasons were loosening (43%), infection (31%), stem fracture (6%), limb-length discrepancy (6%), malrotation (6%), and local recurrence (6%). A cortical window was used in 10 of 45 initial revisions (7 cemented, 3 pressfit) including all of the stem fractures, and in 2 of 15 subsequent re-revisions. Cortical fenestration is an effective, bone-preserving method of implant extraction, particularly for broken or cemented stems. It is associated with low rates of loosening and no increase in periprosthetic fractures.


Recovery of Kidney Function in Children Treated with Maintenance Dialysis.
Data on recovery of kidney function in pediatric patients with presumed ESKD are scarce. We examined the occurrence of recovery of kidney function and its determinants in a large cohort of pediatric patients on maintenance dialysis in Europe. Data for 6574 patients from 36 European countries commencing dialysis at an age below 15 years, between 1990 and 2014 were extracted from the European Society for Pediatric Nephrology/European Renal Association-European Dialysis and Transplant Association Registry. Recovery of kidney function was defined as discontinuation of dialysis for at least 30 days. Time to recovery was studied using a cumulative incidence competing risk approach and adjusted Cox proportional hazard models. Two years after dialysis initiation, 130 patients (2%) experienced recovery of their kidney function after a median of 5.0 (interquartile range, 2.0-9.6) months on dialysis. Compared with patients with congenital anomalies of the kidney and urinary tract, recovery more often occurred in patients with vasculitis (11% at 2 years; adjusted hazard ratio [HR], 20.4; 95% confidence interval [95% CI], 9.7 to 42.8), ischemic kidney failure (12%; adjusted HR, 11.4; 95% CI, 5.6 to 23.1), and hemolytic uremic syndrome (13%; adjusted HR, 15.6; 95% CI, 8.9 to 27.3). Younger age and initiation on hemodialysis instead of peritoneal dialysis were also associated with recovery. For 42 patients (32%), recovery was transient as they returned to kidney replacement therapy after a median recovery period of 19.7 (interquartile range, 9.0-41.3) months. We demonstrate a recovery rate of 2% within 2 years after dialysis initiation in a large cohort of pediatric patients on maintenance dialysis. There is a clinically important chance of recovery in patients on dialysis with vasculitis, ischemic kidney failure, and hemolytic uremic syndrome, which should be considered when planning kidney transplantation in these children.


Raising the bar for the care of seriously ill patients: results of a national survey to define essential palliative care competencies for medical students and residents.
Given the shortage of palliative care specialists in the United States, to ensure quality of care for patients with serious, life-threatening illness, generalist-level palliative care competencies need to be defined and taught. The purpose of this study was to define essential competencies for medical students and internal medicine and family medicine (IM/FM) residents through a national survey of palliative care experts. Proposed competencies were derived from existing hospice and palliative medicine fellowship competencies and revised to be developmentally appropriate for students and residents. In spring 2012, the authors administered a Web-based, national cross-sectional survey of palliative care educational experts to assess ratings and rankings of proposed competencies and competency domains. The authors identified 18 comprehensive palliative care competencies for medical students and IM/FM residents, respectively. Over 95% of survey respondents judged the competencies as comprehensive and developmentally appropriate (survey response rate = 72%, 71/98). Using predefined cutoff criteria, experts identified 7 medical student and 13 IM/FM resident competencies as essential. Communication and pain/symptom management were rated as the most critical domains. This national survey of palliative care experts defines comprehensive and essential palliative care competencies for medical students and IM/FM residents that are specific, measurable, and can be used to report educational outcomes; provide a sequence for palliative care curricula in undergraduate and graduate medical education; and highlight the importance of educating medical trainees in communication and pain management. Next steps include seeking input and endorsement from stakeholders in the broader medical education community.


Glial cells are increased proportionally in transgenic optic nerves with increased numbers of axons.
To study how an increase in axon number influences the number of glial cells in the mammalian optic nerve, we have analyzed a previously described transgenic mouse that expresses the human bcl-2 gene from a neuron-specific enolase promoter. In these mice, the normal postnatal loss of retinal ganglion cell axons is greatly decreased and, as a consequence, the number of axons in the optic nerve is increased by approximately 80% compared with wild-type mice. Remarkably, the numbers of oligodendrocytes, astrocytes, and microglial cells are all increased proportionally in the transgenic optic nerve. The increase in oligodendrocytes apparently results from both a decrease in normal oligodendrocyte death and an increase in oligodendrocyte precursor cell proliferation, whereas the increase in astrocytes apparently results from an increase in the proliferation of astrocyte lineage cells. Unexpectedly, the transgene is expressed in oligodendrocytes and astrocytes, but this does not seem to be responsible for the increased numbers of these cells. These findings indicate that developing neurons and glial cells can interact to adjust glial cell numbers appropriately when neuronal numbers are increased. We also show that the expression of the bcl-2 transgene in retinal ganglion cells protects the cell body from programmed cell death when the axon is cut, but it does not protect the isolated axon from Wallerian degeneration, even though the transgene-encoded protein is present in the axon.


An outcome study of severe traumatic head injury using the "Lund therapy" with low-dose prostacyclin.
There are two independent head injury outcome studies using the "Lund concept", and both showed a mortality rate of about 10%, and a favourable outcome (Glasgow outcome scale, GOS 4 and 5) of about 70%. The Lund concept aims at controlling intracranial pressure, and improving microcirculation around contusions. Intracranial pressure is controlled by maintaining a normal colloid osmotic pressure and reducing the hydrostatic capillary pressure. Microcirculation is improved by ensuring strict normovolaemia and reducing sympathetic discharge. The endogenous substance prostacyclin with its antiaggregatory/antiadhesive effects may further improve microcirculation, which finds support from a microdialysis-based clinical study and an experimental brain trauma study. The present clinical outcome study aims at evaluating whether the previously obtained good outcome with the Lund therapy can be reproduced, and whether the addition of prostacyclin has any adverse side-effects. All 31 consecutive patients with severe head injury, Glasgow coma scale (GCS) < or = 8, admitted to the University Hospital of Umeå during 1998 were included. The Lund therapy including prostacyclin infusion for the first three days at a dose of 0.5 ng kg(-1) min(-1). Outcome was evaluated according to the GOS >10 months after the injury. One patient died, another suffered vegetative state and 7 severe disability. Of the 22 patients with favourable outcome, 19 showed good recovery and 3 moderate disability. No adverse side-effects of prostacyclin were observed. The outcome results from previous studies using the Lund therapy were reproduced, and no adverse side-effects of low-dose prostacyclin were observed.


Further insights into the catalytical properties of deglycosylated pyranose dehydrogenase from Agaricus meleagris recombinantly expressed in Pichia pastoris.
The present study focuses on fragmented deglycosylated pyranose dehydrogenase (fdgPDH) from Agaricus meleagris recombinantly expressed in Pichia pastoris . Fragmented deglycosylated PDH is formed from the deglycosylated enzyme (dgPDH) when it spontaneously loses a C-terminal fragment when stored in a buffer solution at 4 °C. The remaining larger fragment has a molecular weight of ∼46 kDa and exhibits higher volumetric activity for glucose oxidation compared with the deglycosylated and glycosylated (gPDH) forms of PDH. Flow injection amperometry and cyclic voltammetry were used to assess and compare the catalytic activity of the three investigated forms of PDH, "wired" to graphite electrodes with two different osmium redox polymers: [Os(4,4'-dimethyl-2,2'-bipyridine)2(poly(vinylimidazole))10Cl](+) [Os(dmbpy)PVI] and [Os(4,4'-dimethoxy-2,2'-bipyridine)2(poly-(vinylimidazole))10Cl](+) [Os(dmobpy)PVI]. When "wired" with Os(dmbpy)PVI, the graphite electrodes modified with fdgPDH showed a pronounced increase in the current density with Jmax 13- and 6-fold higher than that observed for gPDH- and dgPDH-modified electrodes, making the fragmented enzyme extraordinarily attractive for further biotechnological applications. An easier access of the substrate to the active site and improved communication between the enzyme and mediator matrix are suggested as the two main reasons for the excellent performance of the fdgPDH when compared with that of gPDH and dgPDH. Three of the four glycosites in PDH: N(75), N(175), and N(252) were assigned using mass spectrometry in conjunction with endoglycosidase treatment and tryptic digestion. Determination of the asparagine residues carrying carbohydrate moieties in PDH can serve as a solid background for production of recombinant enzyme lacking glycosylation.


Contingencies for change in complacent smokers.
The majority of smokers have no plans to quit in the near future. These complacent smokers are less likely to quit than other smokers, and few interventions are known to reduce smoking in this population. Although monetary incentives can reduce complacent smokers' breath carbon monoxide (BCO) levels, it is not clear whether these effects can be sustained beyond the several weeks that past studies have examined. The authors compared complacent smokers randomly assigned to receive incentives for BCO reductions (n=18) or noncontingent incentives (n=19) for 3 months. Contingent incentives were associated with (a) reduced BCO; (b) more BCO samples indicative of abstinence; (c) fewer cigarettes smoked and more days abstinent at study end; and (d) lower salivary cotinine. These behaviors can predict future cessation, and 2 of the 18 smokers (11%) receiving BCO-contingent incentives reported quitting as compared with none in the control group. Contingency management procedures, such as those used here, may effectively promote cessation among complacent smokers and provide a model for understanding the possible effects of some environmental interventions (like workplace smoking bans) on the behavior of complacent smokers.


The bidirectional association between body weight and mobility disability: A population-based cohort.
Obesity is more common in people with mobility disability than in non-disabled individuals, but less is known about the longitudinal effects leading to this health state. To explore the potential bidirectional association between mobility disability and obesity. Participants were identified in the population-based Stockholm Public Health Cohort (2002-2010, n = 17 945). Observations with schizophrenia, depression, eating disorder, or cancer within 5 years during and prior to baseline were excluded. Mobility disability and height and weight to calculate BMI (kg/m(2)) were self-reported. We used multivariate-adjusted regression models to estimate relative risks (RRs) and 95% confidence intervals (CI) based on new cases of mobility disability in cohorts that were obese, overweight or normal weight at baseline, and increases in BMI over time by mobility disability status. Obesity at baseline was associated with incident mobility disability. The highest risk was observed in middle-aged women (RR = 3.95, CI = 2.35-6.65). Young men and middle-aged women with long-term mobility disability increased more in BMI (men: 1.55 kg/m(2), 0.61-2.49; women: 0.38, 0.01-0.75), as well as young and middle-aged people with incident mobility disability (young men: 0.68 kg/m(2), 0.10-1.27; middle-aged men: 0.49, 0.20-0.77; young women: 1.41, 0.94-1.87; middle-aged women: 0.64, 0.36-0.93) compared to the groups without any mobility disability. In this paper, we demonstrated the bidirectional and longitudinal associations between body weight and mobility disability and thus the increased risk of developing the combination over time from either condition. Effective health-promotion and prevention strategies are needed to prevent worse health for these vulnerable groups in society.


On the Impact of Electrostatic Correlations on the Double-Layer Polarization of a Spherical Particle in an Alternating Current Field.
At concentrated electrolytes, the ion-ion electrostatic correlation effect is considered an important factor in electrokinetics. In this paper, we compute, in theory and simulation, the dipole moment for a spherical particle (charged, dielectric) under the action of an alternating electric field using the modified continuum Poisson-Nernst-Planck (PNP) model by Bazant et al. [ Double Layer in Ionic Liquids: Overscreening Versus Crowding . Phys. Rev. Lett. 2011 , 106 , 046102 ] We investigate the dependency of the dipole moment in terms of frequency and its variation with such quantities like ζ-potential, electrostatic correlation length, and double-layer thickness. With thin electric double layers, we develop simple models through performing an asymptotic analysis of the modified PNP model. We also present numerical results for an arbitrary Debye screening length and electrostatic correlation length. From the results, we find a complicated impact of electrostatic correlations on the dipole moment. For instance, with increasing the electrostatic correlation length, the dipole moment decreases and reaches a minimum and then it goes up. This is because of initially decreasing of surface conduction and finally increasing due to the impact of ion-ion electrostatic correlations on ion's convection and migration. Also, we show that in contrast to the standard PNP model, the modified PNP model can qualitatively explain the data from the experimental results in multivalent electrolytes.


iFORM/eQTL: an ultrahigh-dimensional platform for inferring the global genetic architecture of gene transcripts.
Knowledge about how changes in gene expression are encoded by expression quantitative trait loci (eQTLs) is a key to construct the genotype-phenotype map for complex traits or diseases. Traditional eQTL mapping is to associate one transcript with a single marker at a time, thereby limiting our inference about a complete picture of the genetic architecture of gene expression. Here, we implemented an ultrahigh-dimensional variable selection model to build a computing platform that can systematically scan main effects and interaction effects among all possible loci and identify a set of significant eQTLs modulating differentiation and function of gene expression. This platform, named iFORM/eQTL, was assembled by forward-selection-based procedures to tackle complex covariance structures of gene-gene interactions. iFORM/eQTL can particularly discern the role of cis-QTLs, trans-QTLs and their epistatic interactions in gene expression. Results from the reanalysis of a published genetic and genomic data set through iFORM/eQTL gain new discoveries on the genetic origin of gene expression differentiation in Caenorhabditis elegans, which could not be detected by a traditional one-locus/one-transcript analysis approach.


Impact of Glutamate Carboxylation in the Adsorption of the α-1 Domain of Osteocalcin to Hydroxyapatite and Titania.
One proposed mechanism of implant fouling is attributed to the nonspecific adsorption of non-collagenous bone matrix proteins (NCPs) onto a newly implanted interface. With the goal of capturing the fundamental mechanistic and thermodynamic forces that govern changes in these NCP recognition domains as a function of γ-carboxyglutamic acid (Gla) post-translational modification and surface chemistry, we probe the adsorption process of the most commonly occurring NCP, osteocalcin, onto a mineral and metal oxide surface. Here, we apply two enhanced sampling methods to independently probe the effects of post-translational modification and peptide structure on adsorption. First, well-tempered metadynamics was used to capture the binding of acetyl and N-methylamide capped glutamic acid and Gla single amino acids onto crystalline hydroxyapatite and titania model surfaces at physiological pH. Following this, parallel tempering metadynamics in the well-tempered ensemble (PTMetaD-WTE) was used to study adsorption of the α-1 domain of osteocalcin onto hydroxyapatite and titania. Simulations were performed for the α-1 domain of osteocalcin in both its fully decarboxylated (dOC) and fully carboxylated (OC) form. Our simulations find that increased charge density due to carboxylation results in increased interactions at the interface, and stronger adsorption of the single amino acids to both surfaces. Interestingly, the role of Gla in promoting compact and helical structure in the α-1 domain resulted in disparate binding modes at the two surfaces, which is attributed to differences in interfacial water behavior. Overall, this work provides a benchmark for understanding the mechanisms that drive adsorption of Gla-containing mineralizing proteins onto different surface chemistries.


Structural basis underlying viral hijacking of a histone chaperone complex.
The histone H3.3 chaperone DAXX is implicated in formation of heterochromatin and transcription silencing, especially for newly infecting DNA virus genomes entering the nucleus. Epstein-Barr virus (EBV) can efficiently establish stable latent infection as a chromatinized episome in the nucleus of infected cells. The EBV tegument BNRF1 is a DAXX-interacting protein required for the establishment of selective viral gene expression during latency. Here we report the structure of BNRF1 DAXX-interaction domain (DID) in complex with DAXX histone-binding domain (HBD) and histones H3.3-H4. BNRF1 DID contacts DAXX HBD and histones through non-conserved loops. The BNRF1-DAXX interface is responsible for BNRF1 localization to PML-nuclear bodies typically associated with host-antiviral resistance and transcriptional repression. Paradoxically, the interface is also required for selective transcription activation of viral latent cycle genes required for driving B-cell proliferation. These findings reveal molecular details of virus reprogramming of an antiviral histone chaperone to promote viral latency and cellular immortalization.


Effect of Thermal and Shear Stressors on the Physical Properties, Structural Integrity and Biological Activity of the Anti-TNF-alpha Monoclonal Antibody, Infliximab.
The influence of thermal and shear stressors on the stability of the anti-TNF-α monoclonal antibody (mAb), Infliximab® (INF) was investigated. INF at concentrations of 1, 4 and 10 mg/ml was subjected to thermal stress at temperatures of 25-65°C and to shear force by sonication for 1 and 3 minutes. The stressed samples were analysed for physical properties by particle size, zeta potential, for structural integrity by gel electrophoresis (SDS-PAGE) and circular dichroism, INF content by UV spectroscopy and for biological activity by ELISA. Results show no change in physical properties or structural integrity of INF at any concentration tested, when subjected to a temperature of up to 50°C. At 65°C, aggregation and precipitation of INF was observed. When subjected to shear stress, higher concentrations of INF at 4 and 10mg/ml maintained their physical properties and structural integrity. However, the biological activity of INF was found to decrease with increasing temperature and sonication time, and was concentration dependent (ANOVA; p<0.05). Interestingly, lyophilisation of INF at 1mg/ml did not affect its physical properties, structural integrity or its biological activity. These findings have important implications with respect to pharmaceutical processing of INF and mAbs including formulation as polymeric micro and nanoparticle systems for sustained or targeted delivery. These findings also have important implications with respect to the handling and storage of INF and mAbs for clinical use.


A biomechanical comparison of subscapularis repair techniques in total shoulder arthroplasty: lesser tuberosity osteotomy versus subscapularis peel.
The subscapularis peel (SP) and the lesser tuberosity osteotomy (LTO) are 2 common exposure techniques for total shoulder arthroplasty. Although some biomechanical studies have suggested a higher resistance to failure with the LTO, clinical studies have demonstrated no difference in repair failure or tendon healing. We hypothesized that there would be no difference in biomechanically tested repair strength between our SP technique and the previously tested LTO technique. Eleven cadaver shoulders were separated into 2 groups: 6 SPs and 5 LTOs. After initial loading for 3000 cycles, the specimens were incrementally loaded to 450 ± 50 N or catastrophic failure. Repair gapping was measured after cyclical loading, and fatigue life was analyzed after incremental loading. There was no significant difference in mean repair gapping between the SP (2.40 ± 0.36 mm; mean ± standard deviation) and the LTO groups (3.10 ± 2.93 mm; P = .57). There was also no difference in the mean number of cycles to failure (6894 ± 956 vs. 6018 ± 1179; P = .14) and mean load to failure (400 ± 79 N vs. 340 ± 91 N; P = .21) between the SP and LTO techniques. However, there was more variability in bead gapping in the LTO group (P < .01). No significant differences were found in repair gapping, fatigue failure, and load to failure in comparing the SP and LTO repairs. However, the SP repair demonstrated significantly less variability in repair gapping. These findings suggest that initial fixation biomechanical properties between the 2 constructs are similar in vitro.


Migration, acculturation, and the maintenance of between-group cultural variation.
How do migration and acculturation (i.e. psychological or behavioral change resulting from migration) affect within- and between-group cultural variation? Here I address this question by drawing analogies between genetic and cultural evolution. Population genetic models show that migration rapidly breaks down between-group genetic structure. In cultural evolution, however, migrants or their descendants can acculturate to local behaviors via social learning processes such as conformity, potentially preventing migration from eliminating between-group cultural variation. An analysis of the empirical literature on migration suggests that acculturation is common, with second and subsequent migrant generations shifting, sometimes substantially, towards the cultural values of the adopted society. Yet there is little understanding of the individual-level dynamics that underlie these population-level shifts. To explore this formally, I present models quantifying the effect of migration and acculturation on between-group cultural variation, for both neutral and costly cooperative traits. In the models, between-group cultural variation, measured using F statistics, is eliminated by migration and maintained by conformist acculturation. The extent of acculturation is determined by the strength of conformist bias and the number of demonstrators from whom individuals learn. Acculturation is countered by assortation, the tendency for individuals to preferentially interact with culturally-similar others. Unlike neutral traits, cooperative traits can additionally be maintained by payoff-biased social learning, but only in the presence of strong sanctioning mechanisms (e.g. institutions). Overall, the models show that surprisingly little conformist acculturation is required to maintain realistic amounts of between-group cultural diversity. While these models provide insight into the potential dynamics of acculturation and migration in cultural evolution, they also highlight the need for more empirical research into the individual-level learning biases that underlie migrant acculturation.


Hypoxia-directed and activated theranostic agent: Imaging and treatment of solid tumor.
Hypoxia, a distinguished feature of various solid tumors, has been considered as a key marker for tumor progression. Inadequate vasculature and high interstitial pressures result in relatively poor drug delivery to these tumors. Herein, we developed an antitumor theranostic agent, 4, which is activated in hypoxic conditions and can be used for the diagnosis and treatment of solid tumors. Compound 4, bearing biotin, a tumor-targeting unit, and SN38, an anticancer drug, proved to be an effective theranostic agent for solid tumors. SN38 plays a dual role: as an anticancer drug for therapy and as a fluorophore for diagnosis, thus avoids an extra fluorophore and limits cytotoxicity. Compound 4, activated in the hypoxic environment, showed high therapeutic activity in A549 and HeLa cells and spheroids. In vivo imaging of solid tumors confirmed the tumor-specific localization, deep tissue penetration and activation of compound 4, as well as the production of a strong anticancer effect through the inhibition of tumor growth in a xenograft mouse model validating it as a promising strategy for the treatment of solid tumors.


More about polyhedra-interpretation through constructions in the image plane.
This paper deals with the interpretation and feasibility check of line drawings representing polyhedral scenes. The polyhedra are of general types and there are no restrictions on camera position. The geometric consistency check and the line labeling are carried out through constructions in the image plane. An algorithm for the geometric construction is suggested, and the necessary conditions for these constructions are discussed. The image plane construction can be used for preparing labeled junction catalogs for junctions other than trihedral. In addition the paper analyzes the relation between the image plane construction and the gradient space construction suggested by Mackworth [7] for the same purpose.


The Effects of Auricular and Body Acupuncture in Turkish Obese Female Patients: A Randomized Controlled Trial Indicated Both Methods Lost Body Weight But Auricular Acupuncture Was Better Than Body Acupuncture.
It was aimed to observe the efficacy of body acupuncture and auricular acupuncture for the management of obesity. Fifty female obese patients were randomized into 2 groups; a Body Acupuncture Group (GBA; n=25) and an Auricular Acupuncture Group (GAA; n=25). All patients were followed up for 12 weeks. Their anthropometric measurements (body weight, body mass index, body fat percentage, waist circumference and hip circumference) were recorded for analysis in every 15 days. Patients were compared in terms of anthropometric measurements. Patients in the GAA received 6 sessions of auricular acupuncture in every 15 days. We used bilateral 2 points namely anti-aggression point and stomach point for auricular acupuncture. Bilateral L14, LIll, ST25, ST36, SP6, SP9, CV12, and CV6 points were selected for two times in a week (totally 24 sessions) in the GBA. Independent samples t test, patients in GAA completed the study. The mean age of patients was 36.2±10.5 years (GBA: 32.7±12.3; GAA: 39.1±7.9). The mean weight was 95.7±11.8 kg before acupuncture, and it decreased to 92.5±14.4 kg after acupuncture in GBA; and it was 95.1±15.6 kg before acupuncture and it decreased to 91.5±11.0 kg after acupuncture in GAA. Change in weight after acupuncture was 2.6±2.4 kg in GBA, and 4.2±3.4 kg in GAA (p<0.05). In both groups, changes in weight, body mass index, body fat percentage, waist circumference, and hip circumference were statistically significant (p<0.05). Only one patient reported side effect related to acupuncture. Body acupuncture and auricular acupuncture have the therapeutic role in the management of obesity. In this study, auricular acupuncture is more effective in reducing body weight than body acupuncture. Thus, auricular may be useful and safe, non-pharmacological treatment.


Electrochemically switchable rotaxanes: recent strides in new directions.
Are they still electrifying? Electrochemically switchable rotaxanes are well known for their ability to efficiently undergo changes of (co-)conformation and properties under redox-control. Thus, these mechanically interlocked assemblies represent an auspicious liaison between the fields of molecular switches and molecular electronics. Since the first reported example of a redox-switchable molecular shuttle in 1994, improved tools of organic and supramolecular synthesis have enabled sophisticated new architectures, which provide precise control over properties and function. This perspective covers recent advances in the area of electrochemically active rotaxanes including novel molecular switches and machines, metal-containing rotaxanes, non-equilibrium systems and potential applications.


Deletions at the SOX10 gene locus cause Waardenburg syndrome types 2 and 4.
Waardenburg syndrome (WS) is an auditory-pigmentary disorder that exhibits varying combinations of sensorineural hearing loss and abnormal pigmentation of the hair and skin. Depending on additional symptoms, WS is classified into four subtypes, WS1-WS4. Absence of additional features characterizes WS2. The association of facial dysmorphic features defines WS1 and WS3, whereas the association with Hirschsprung disease (aganglionic megacolon) characterizes WS4, also called "Waardenburg-Hirschsprung disease." Mutations within the genes MITF and SNAI2 have been identified in WS2, whereas mutations of EDN3, EDNRB, and SOX10 have been observed in patients with WS4. However, not all cases are explained at the molecular level, which raises the possibility that other genes are involved or that some mutations within the known genes are not detected by commonly used genotyping methods. We used a combination of semiquantitative fluorescent multiplex polymerase chain reaction and fluorescent in situ hybridization to search for SOX10 heterozygous deletions. We describe the first characterization of SOX10 deletions in patients presenting with WS4. We also found SOX10 deletions in WS2 cases, making SOX10 a new gene of WS2. Interestingly, neurological phenotypes reminiscent of that observed in WS4 (PCWH syndrome [peripheral demyelinating neuropathy, central dysmyelinating leukodystrophy, WS, and Hirschsprung disease]) were observed in some WS2-affected patients with SOX10 deletions. This study further characterizes the molecular complexity and the close relationship that links the different subtypes of WS.


Shift-invariant three-dimensional object recognition by means of digital holography.
We present an optoelectronic method to analyze three-dimensional (3D) scenes that is able to detect the presence, and also the position and orientation, of a reference 3D object. The data-acquisition procedure is based on digital holography. A phase-shifting interferometer records a single digital Fresnel hologram of the 3D scene with an intensity-recording device. Holographic information of the 3D reference object is also obtained with the same method. Correlation techniques are then applied to recognize the presence and position of the 3D reference object in the 3D scene. The technique also allows us to detect the 3D reference with a small out-of-plane rotation. Preliminary experimental results are presented that demonstrate the theory.


Concordant and discordant alcohol, tobacco, and marijuana use as predictors of marital dissolution.
This study examined concordant and discrepant alcohol, tobacco, and marijuana use among couples to determine whether they predicted marital separation or divorce over 9 years. The study recruited 634 couples as they applied for their marriage licenses; we assessed them at that time and reassessed them with mailed questionnaires at their first, second, fourth, seventh, and ninth wedding anniversaries. Approximately 60% of the men and women were European American, and approximately 33% were African American. The frequency of drinking to intoxication and binge drinking (more than 5 drinks in an occasion) was assessed, as was the use of cigarettes and marijuana. At each assessment, each member of the couple was asked about the occurrence of marital separations and divorce. Bivariate analyses indicated that tobacco and marijuana use, whether discrepant or concordant, were associated with marital disruptions. However, discrepant heavy drinking was associated with disruptions, but concordant heavy drinking was not. Concordant and discordant marijuana use were not associated with divorce when analyses controlled for alcohol and tobacco use. Concordant and discordant tobacco use was not associated with divorce when analyses controlled for sociodemographic and personality factors. However, discrepant alcohol use was related to divorce after controlling for the other substances in 1 analysis and after controlling for the sociodemographic factors in a separate analysis. Tobacco and marijuana use were related to divorce through their associations with other variables. However, results suggest that discrepant alcohol use may lead to marital disruptions and should be addressed with couples seeking marital treatment.


The Mediterranean diet and incidence of hypertension: the Seguimiento Universidad de Navarra (SUN) Study.
The Mediterranean diet is receiving increasing attention in cardiovascular epidemiology. The association of adherence to the Mediterranean diet with the incidence of hypertension was evaluated among 9,408 men and women enrolled in a dynamic Spanish prospective cohort study during 1999-2005. Dietary intake was assessed at baseline with a validated semiquantitative food frequency questionnaire, and a 9-point Mediterranean diet score was constructed. During a median follow-up period of 4.2 years (range, 1.9-7.9), 501 incident cases of hypertension were identified. After adjustment for major hypertension risk factors and nutritional covariates, adherence to the Mediterranean diet was not associated with hypertension (the hazard ratio was 1.10 (95% confidence interval (CI): 0.81, 1.41) for moderate adherence and 1.12 (95% CI: 0.79, 1.60) for high adherence). However, it was associated with reduced changes in mean levels of systolic blood pressure (moderate adherence, -2.4 mm Hg (95% CI: -4.0, -0.8); high adherence, -3.1 mm Hg (95% CI: -5.4, -0.8)) and diastolic blood pressure (moderate adherence, -1.3 mm Hg (95% CI: -2.5, -0.1); high adherence, -1.9 mm Hg (95% CI: -3.6, -0.1)) after 6 years of follow-up. These results suggest that adhering to a Mediterranean-type diet could contribute to the prevention of age-related changes in blood pressure.


Mental Toughness in South African Youth.
Young people are particularly vulnerable to health risk behaviors and interpersonal violence, stimulating scholars' attention towards identifying factors that may reduce the likelihood that these actions will occur. Associated with positive outcomes in a variety of domains, mental toughness in young people might protect them from engaging in potentially deleterious interpersonal or health-risk behaviors, while potentially promoting positive psychological behaviors. Within this framework, the present study investigated the relationships between mental toughness, attitudes towards physical and psychological risk-taking, and trait forgiveness in a sample of 123 (males = 54, females = 69) South African youth ( M age = 23.97 years, SD = 4.46). Univariate and multivariate analyses indicated higher levels of mental toughness were associated with being more forgiving, ([Formula: see text] = .036), perceiving physical risk-taking more positively ([Formula: see text] = .062), but having more negative attitudes towards psychological risk-taking ([Formula: see text] = .036). These findings give credence to mental toughness as a psychological characteristic involved in youth risk-taking perceptions and interpersonal functioning. Future research might explore the integration of mental toughness into the development of future youth risk behavior interventions.


ALS disrupts spinal motor neuron maturation and aging pathways within gene co-expression networks.
Modeling amyotrophic lateral sclerosis (ALS) with human induced pluripotent stem cells (iPSCs) aims to reenact embryogenesis, maturation and aging of spinal motor neurons (spMNs) in vitro. As the maturity of spMNs grown in vitro compared to spMNs in vivo remains largely unaddressed, it is unclear to what extent this in vitro system captures critical aspects of spMN development and molecular signatures associated with ALS. Here, we compared transcriptomes among iPSC-derived spMNs, fetal spinal tissues and adult spinal tissues. This approach produced a maturation scale revealing that iPSC-derived spMNs were more similar to fetal spinal tissue than to adult spMNs. Additionally, we resolved gene networks and pathways associated with spMN maturation and aging. These networks enriched for pathogenic familial ALS genetic variants and were disrupted in sporadic ALS spMNs. Altogether, our findings suggest that developing strategies to further mature and age iPSC-derived spMNs will provide more effective iPSC models of ALS pathology.


MxiG, a membrane protein required for secretion of Shigella spp. Ipa invasins: involvement in entry into epithelial cells and in intercellular dissemination.
Entry of Shigella flexneri into epithelial cells involves secretory proteins, the Ipa proteins, and their dedicated secretion apparatus, the Mxi-Spa translocon, which is encoded by the mxi and spa operons. We have characterized the mxiG gene that is located at the proximal part of the mxi operon. Inactivation of mxiG abolished lpa secretion, which indicates that MxiG is an essential component of the Mxi-Spa translocon. Immunoblotting analysis of membrane fractions suggests that the 42 kDa MxiG protein is associated with both the inner and outer membranes. Taking advantage of the complementation of the mxiG mutant by a plasmid carrying a wild-type copy of mxiG (which restored Ipa secretion, entry into HeLa cells, and cell-to-cell spread) we mutagenized the mxiG gene carried by the complementing plasmid to replace the RGD motif of MxiG by RAD. This mutation (mxiG*), which had no effect on the stability of the protein, did not affect Ipa secretion in vitro or entry into HeLa cells, but impaired intercellular dissemination. Therefore, MxiG and possibly proteins secreted by the Mxi-Spa translocation are involved not only in entry but also in spread of Shigella between epithelial cells.


Dose-response study of effect of oleuropein, an olive oil polyphenol, in an ovariectomy/inflammation experimental model of bone loss in the rat.
This study was carried out to assess the dose-dependent bone-sparing effect of oleuropein, an olive oil phenolic compound with anti-inflammatory and anti-oxidative properties, on bone loss induced by talc granulomatosis in oestrogen-deficient rat. Among 98 rats, 20 were sham-operated (SH) while the others (78) were ovariectomised (OVX). The SH and 26 OVX rats (controls) were given a standard diet for 100 days. The 52 remaining OVX rats were allocated to 4 groups that received oleuropein at 2.5, 5, 10 or 15 mg/kg body weight per day for 100 days. Three weeks before necropsy, an inflammation was induced by subcutaneous injections of talc in half of the SH and OVX rats and in all oleuropein-treated animals. Castration was associated with a decreased bone mineral density (BMD). In OVX rats, inflammation, characterised by an increase of the spleen weight and plasma fibrinogen levels, exacerbated this bone loss, as shown by values of BMD of the total femur metaphyseal and diaphyseal subregions. The 4 doses of oleuropein reduced bone loss and improved inflammatory biomarkers excepted for 5mg/kg BW. Every dose of oleuropein elicited protective effects on bone mass in this model of ovariectomy associated with inflammation, probably by modulating inflammatory parameters.


The analysis of Arabidopsis nicotianamine synthase mutants reveals functions for nicotianamine in seed iron loading and iron deficiency responses.
Nicotianamine chelates and transports micronutrient metal ions in plants. It has been speculated that nicotianamine is involved in seed loading with micronutrients. A tomato (Solanum lycopersicum) mutant (chloronerva) and a tobacco (Nicotiana tabacum) transgenic line have been utilized to analyze the effects of nicotianamine loss. These mutants showed early leaf chlorosis and had sterile flowers. Arabidopsis (Arabidopsis thaliana) has four NICOTIANAMINE SYNTHASE (NAS) genes. We constructed two quadruple nas mutants: one had full loss of NAS function, was sterile, and showed a chloronerva-like phenotype (nas4x-2); another mutant, with intermediate phenotype (nas4x-1), developed chlorotic leaves, which became severe upon transition from the vegetative to the reproductive phase and upon iron (Fe) deficiency. Residual nicotianamine levels were sufficient to sustain the life cycle. Therefore, the nas4x-1 mutant enabled us to study late nicotianamine functions. This mutant had no detectable nicotianamine in rosette leaves of the reproductive stage but low nicotianamine levels in vegetative rosette leaves and seeds. Fe accumulated in the rosette leaves, while less Fe was present in flowers and seeds. Leaves, roots, and flowers showed symptoms of Fe deficiency, whereas leaves also showed signs of sufficient Fe supply, as revealed by molecular-physiological analysis. The mutant was not able to fully mobilize Fe to sustain Fe supply of flowers and seeds in the normal way. Thus, nicotianamine is needed for correct supply of seeds with Fe. These results are fundamental for plant manipulation approaches to modify Fe homeostasis regulation through alterations of NAS genes.


Systematic approach to musculoskeletal benign tumors.
The radiologic workup of musculoskeletal tumors can be both cost-efficient and extremely helpful to the referring clinician if one proceeds in a thoughtful and logical manner. It should be remembered that plain films remain the most reliable imaging method for assessment of both biological activity and probable histologic diagnosis of an osseous lesion. Further investigations are of help to determine the extent of lesion and to help in staging. In order to do this, we have found it useful to include an assessment of 10 determinants in the description of a tumor. If these determinants are accurately described, the correct diagnosis or at least a limited differential diagnosis usually becomes obvious.


Modelling the potential population impact and cost-effectiveness of self-testing for HIV: evaluation of data requirements.
HIV testing uptake has increased dramatically in recent years in resource limited settings. Nevertheless, over 50% of the people living with HIV are still unaware of their status. HIV self-testing (HIVST) is a potential new approach to facilitate further uptake of testing which requires consideration, taking into account economic factors. Mathematical models and associated economic analysis can provide useful assistance in decision-making processes, offering insight, in this case, into the potential long-term impact at a population level and the price-point at which free or subsidized HIVST would be cost-effective in a given setting. However, models are based on assumptions, and if the required data are sparse or limited, this uncertainty will be reflected in the results from mathematical models. The aim of this paper is to describe the issues encountered in modeling the cost-effectiveness of introducing HIVST, to indicate the evidence needed to support various modeling assumptions, and thus which data on HIVST would be most beneficial to collect.


Mutations in human monoamine-related neurotransmitter pathway genes.
Biosynthesis and metabolism of serotonin and catecholamines involve at least eight individual enzymes that are mainly expressed in tissues derived from the neuroectoderm, e.g., the central nervous system (CNS), pineal gland, adrenal medulla, enterochromaffin tissue, sympathetic nerves, and ganglia. Some of the enzymes appear to have additional biological functions and are also expressed in the heart and various other internal organs. The biosynthetic enzymes are tyrosine hydroxylase (TH), tryptophan hydroxylases type 1 and 2 (TPH1, TPH2), aromatic amino acid decarboxylase (AADC), dopamine beta-hydroxylase (DbetaH), and phenylethanolamine N-methyltransferase (PNMT), and the specific catabolic enzymes are monoamine oxidase A (MAO-A) and catechol O-methyltransferase (COMT). For the TH, DDC, DBH, and MAOA genes, many single nucleotide polymorphisms (SNPs) with unknown function, and small but increasing numbers of cases with autosomal recessive mutations have been recognized. For the remaining genes (TPH1, TPH2, PNMT, and COMT) several different genetic markers have been suggested to be associated with regulation of mood, pain perception, and aggression, as well as psychiatric disturbances such as schizophrenia, depression, suicidality, and attention deficit/hyperactivity disorder. The genetic markers may either have a functional role of their own, or be closely linked to other unknown functional variants. In the future, molecular testing may become important for the diagnosis of such conditions. Here we present an overview on mutations and polymorphisms in the group of genes encoding monoamine neurotransmitter metabolizing enzymes. At the same time we propose a unified nomenclature for the nucleic acid aberrations in these genes. New variations or details on mutations will be updated in the Pediatric Neurotransmitter Disorder Data Base (PNDDB) database (www.bioPKU.org).


Long-term field performance of biosand filters in the Artibonite Valley, Haiti.
A field study assessing the sustainability and efficacy of 55 biosand filters installed during 1999-2010 was conducted in the Artibonite Valley, Haiti during 2011. Twenty-nine filters were still in use. Duration of filter use ranged from < 1 to 12 years. Water quality, microbial analysis, and flow rate were evaluated for each functioning filter. Kaplan-Meier analysis of filter lifespans showed that filter use remained high (> 85%) up to seven years after installation. Several filters were still in use after 12 years, which is longer than documented in any previous study. Filtered water from 25 filters (86%) contained Escherichia coli concentrations of < 10 most probable number of coliforms/100 mL. Recontamination of stored filtered water was negligible. Bacterial removal efficiency was 1.1 log(10). Comparable results from previous studies in the same region and elsewhere show that biosand filter technology continues to be an effective and sustainable water treatment method in developing countries worldwide.


Whole-Genome Characterization of Epidemic Neisseria meningitidis Serogroup C and Resurgence of Serogroup W, Niger, 2015.
In 2015, Niger reported the largest epidemic of Neisseria meningitidis serogroup C (NmC) meningitis in sub-Saharan Africa. The NmC epidemic coincided with serogroup W (NmW) cases during the epidemic season, resulting in a total of 9,367 meningococcal cases through June 2015. To clarify the phylogenetic association, genetic evolution, and antibiotic determinants of the meningococcal strains in Niger, we sequenced the genomes of 102 isolates from this epidemic, comprising 81 NmC and 21 NmW isolates. The genomes of 82 isolates were completed, and all 102 were included in the analysis. All NmC isolates had sequence type 10217, which caused the outbreaks in Nigeria during 2013-2014 and for which a clonal complex has not yet been defined. The NmC isolates from Niger were substantially different from other NmC isolates collected globally. All NmW isolates belonged to clonal complex 11 and were closely related to the isolates causing recent outbreaks in Africa.


Mesozoic retroposons reveal parrots as the closest living relatives of passerine birds.
The relationships of passerines (such as the well-studied zebra finch) with non-passerine birds is one of the great enigmas of avian phylogenetic research, because decades of extensive morphological and molecular studies yielded highly inconsistent results between and within data sets. Here we show the first application of the virtually homoplasy-free retroposon insertions to this controversy. Our study examined ~200,000 retroposon-containing loci from various avian genomes and retrieved 51 markers resolving early bird phylogeny. Among these, we obtained statistically significant evidence that parrots are the closest and falcons the second-closest relatives of passerines, together constituting the Psittacopasserae and the Eufalconimorphae, respectively. Our new and robust phylogenetic framework has substantial implications for the interpretation of various conclusions drawn from passerines as model organisms. This includes insights of relevance to human neuroscience, as vocal learning (that is, birdsong) probably evolved in the psittacopasseran ancestor, >30 million years earlier than previously assumed.


Neurodisability among Children at the Nexus of the Child Welfare and Youth Justice System.
Although neurodisability features significantly across child welfare and youth justice cohorts, little research investigates neurodisability among crossover children with dual systems involvement. This study examined differences in childhood adversity, child protection involvement, and offending among crossover children by neurodisability status. Data were from a sample of 300 children (68% male, 31% female, 1% transgender; mean age = 16.2 years, range 10-21) who were charged and appeared in three Australian children's courts, and who also had statutory child protection involvement in the study jurisdiction. The results indicated that nearly one-half of crossover children had a neurodisability (48%) and this group experienced greater cumulative maltreatment and adversity, earlier out-of-home care entry and offending onset, more caregiver relinquishment and residential care placement, and a greater volume of charges. While substantial differences between specific neurodisabilities were evident, crossover children with any neurodisability had greater odds of having charges related to criminal damage and motor vehicle theft, however they were no more likely to have violent charges relative to other crossover children. The study's findings demonstrated that the prevalence of neurodisability, and child welfare system responses to this phenomenon, contributes to several offending-related trends observed among crossover children.


'Neanderthal bone flutes': simply products of Ice Age spotted hyena scavenging activities on cave bear cubs in European cave bear dens.
Punctured extinct cave bear femora were misidentified in southeastern Europe (Hungary/Slovenia) as 'Palaeolithic bone flutes' and the 'oldest Neanderthal instruments'. These are not instruments, nor human made, but products of the most important cave bear scavengers of Europe, hyenas. Late Middle to Late Pleistocene (Mousterian to Gravettian) Ice Age spotted hyenas of Europe occupied mainly cave entrances as dens (communal/cub raising den types), but went deeper for scavenging into cave bear dens, or used in a few cases branches/diagonal shafts (i.e. prey storage den type). In most of those dens, about 20% of adult to 80% of bear cub remains have large carnivore damage. Hyenas left bones in repeating similar tooth mark and crush damage stages, demonstrating a butchering/bone cracking strategy. The femora of subadult cave bears are intermediate in damage patterns, compared to the adult ones, which were fully crushed to pieces. Hyenas produced round-oval puncture marks in cub femora only by the bone-crushing premolar teeth of both upper and lower jaw. The punctures/tooth impact marks are often present on both sides of the shaft of cave bear cub femora and are simply a result of non-breakage of the slightly calcified shaft compacta. All stages of femur puncturing to crushing are demonstrated herein, especially on a large cave bear population from a German cave bear den.


Attributable risk for oral cancer in northern Italy.
Using data from a case-control study conducted between 1984 and 1992 in the provinces of Milan and Pordenone, northern Italy, on 439 cases of oral and pharyngeal cancers and 2106 hospital controls, we computed the population attributable risk for oropharyngeal cancer in relation to tobacco, alcohol, and a measure of low beta-carotene intake. Two different models were used for estimating relative risks, one assuming that the three factors act multiplicatively on the relative risk and the second estimating separately each combination of alcohol and tobacco and assuming a multiplicative model only for beta-carotene. The estimated attributable risks were similar for the two models considered. For both models and both sexes, the single factor with the highest attributable risk was smoking, which accounted for 81-87% of oral cancers in males and for 42-47% in females. Alcohol explained about 60% of male cases, but only 15% of female ones, and low beta-carotene accounted for 24% of total cases (25% of males, 17% of females). Together the three factors were responsible for 91-94% of oropharyngeal cancers in males, 51-57% in females, and 85-88% in both sexes combined. The present knowledge of major identified risk factors could, in principle, reduce the burden of the disease in Italy from 2400 to about 200 deaths per year for males and from 500 to 230 for females, thus explaining the difference in incidence and mortality between the two sexes.


Boosting Secretion of Extracellular Protein by Escherichia coli via Cell Wall Perturbation.
Escherichia coli is one of the most widely used host microorganisms for recombinant protein expression and metabolic engineering, but it cannot efficiently secrete recombinant proteins to extracellular space. Here, extracellular protein secretion was enhanced in E. coli by deleting two d,d-carboxypeptidase genes (dacA and dacB, single and double deletions) to perturb the cell wall peptidoglycan network. Deletion of dacA and dacB enhanced the accumulation of intracellular soluble peptidoglycan in E. coli and affected cell morphology, resulting in a more irregular cell shape and the appearance of transparent bulges. Deletion of dacA and dacB appears to disrupt the normal rigid structure, presumably due to perturbation and destruction of the cell wall peptidoglycan network. The extracellular green fluorescent protein (GFP) fluorescence intensity of deletion mutants was increased by >2.0-fold compared with that of control cells, and that of the double deletion mutant was increased by 2.7-fold. Extracellular recombinant fibroblast growth factor receptor 2 (FGFR2) and collagen E4 secretion in deletion mutants was also enhanced compared with that in the control cells. Additionally, the extracellular recombinant amylase activity of single-deletion mutants BL21 ΔdacA pETDuet-amyk and BL21 ΔdacB pETDuet-amyk was increased 2.5- and 3.1-fold, respectively. The extracellular distribution of α-galactosidase by deletion mutants was also increased by >2.0-fold. Deletion of dacA and dacB increased outer membrane permeability, which could explain the enhanced extracellular protein secretion.IMPORTANCE Cell surface structure stabilization is important for extracellular secretion of proteins in Escherichia coli As the main constituent of the cell wall, peptidoglycan contributes to cell structure robustness and stability. Here, we perturbed the peptidoglycan network by deleting dacA and dacB genes encoding d,d-carboxypeptidase enzymes to improve extracellular protein secretion. This new strategy could enhance the capacity of E. coli as a microbial cell factory for extracellular secretion of proteins and chemicals.


Evidence that neuroleptics increase striatal acetylcholine release through stimulation of dopamine D1 receptors.
The relative role of D1 and D2 dopamine receptors in the neuroleptic-induced increase of striatal acetylcholine (ACh) release was investigated using brain microdialysis in freely moving rats. Administration of (-)-sulpiride, haloperidol and clozapine, produced a dose-related increase in ACh release in the striatum. Maximal increase by 52, 45 and 73% over basal values was produced by the dose of 20, 0.25 and 10 mg/kg i.p. of (-)-sulpiride, haloperidol and clozapine, respectively. Administration of the D1 receptor antagonist SCH 23390 (0.1 mg/kg s.c.) decreased ACh output by 30%, completely suppressed the stimulant effect of (-)-sulpiride and haloperidol and only modestly reduced that of clozapine. The inhibitory effect of SCH 23390 vs. (-)-sulpiride or haloperidol-induced ACh output was shared by SCH 39166 (1 mg/kg i.p.), another specific D1 receptor antagonist. On the other hand, SCH 23390 (0.1 mg/kg s.c.) was ineffective in reducing atropine-induced increase in ACh release. A combined treatment with reserpine (5 mg/kg i.p.) and alpha-methyltyrosine (150 mg/kg i.p.), 6 h beforehand, prevented the enhancement of ACh release induced by both (-)-sulpiride and haloperidol, whereas only reduced that by clozapine. The results indicate that neuroleptics increase striatal ACh release by enhancing endogenous extracellular dopamine acting on D1 receptors, and suggest that these receptors play a major physiological role in controlling ACh release in the striatum.


New advances in erectile technology.
New discoveries and technological advances in medicine are rapid. The role of technology in the treatment of erectile dysfunction (ED) will be widened and more options will be available in the years to come. These erectile technologies include external penile support devices, penile vibrators, low intensity extracorporeal shockwave, tissue engineering, nanotechnology and endovascular technology. Even for matured treatment modalities for ED, such as vacuum erectile devices and penile implants, there is new scientific information and novel technology available to improve their usage and to stimulate new ideas. We anticipate that erectile technologies may revolutionize ED treatment and in the very near future ED may become a curable condition.


Cross-reactivity with bovine cells of monoclonal antibodies submitted to the 6th International Workshop on Human Leukocyte Differentiation Antigens.
Twelve subpanels of monoclonal antibodies (MAb) included within the 6th International Workshop on Human Leukocyte Differentiation Antigens (6th HLDA) were assayed for reactivity with bovine peripheral blood leukocytes. Sixty-nine of the 807 MAb (8.6%) stained bovine cells. These MAb represented 30 different human CD groups. Nine of the MAb to different human CD antigens (CD19, CD23, CD39, CD47, CD86, CD117, CD120b, CDw149, CD165) potentially recognized antigens on cattle cells that had not previously been identified. These were investigated further by two-colour immunofluorescence to compare the cellular expression of the antigen on cattle cells with that reported for the different CD antigens in humans. Four of the MAb that belonged to CD23, CD39, CD47, and CDw149 stained bovine cells in a manner that indicated an almost identical cellular distribution of the antigen to that reported in humans. This implied that these MAb reacted with the homologous cattle molecules. Further work would be necessary to confirm specificity of CD19, CD86, CD117, CD120b and CD165 MAb. Other cross-reacting MAb either recognized antigens already defined in cattle or antigens not yet clustered in humans. The study has identified valuable new reagents for studies of cattle and confirmed that most common cross-reactive MAb are to epitopes on integrins.


Assessing the maximum contribution from ancient populations.
Ancestral relationships between populations separated by time represent an often neglected dimension in population genetics, a field which historically has focused on analysis of spatially distributed samples from the same point in time. Models are usually straightforward when two time-separated populations are assumed to be completely isolated from all other populations. However, this is usually an unrealistically stringent assumption when there is gene flow with other populations. Here, we investigate continuity in the presence of gene flow from unknown populations. This setup allows a more nuanced treatment of questions regarding population continuity in terms of "level of contribution" from a particular ancient population to a more recent population. We propose a statistical framework which makes use of a biallelic marker sampled at two different points in time to assess population contribution, and present two different interpretations of the concept. We apply the approach to published data from a prehistoric human population in Scandinavia (Malmström H, Gilbert MTP, Thomas MG, Brandström M, Storå J, Molnar P, Andersen PK, Bendixen C, Holmlund G, Götherström A, et al. 2009. Ancient DNA reveals lack of continuity between Neolithic hunter-gatherers and contemporary Scandinavians. Curr Biol. 19:1758-1762) and Pleistocene woolly mammoth (Barnes I, Shapiro B, Lister A, Kuznetsova T, Sher A, Guthrie D, Thomas MG. 2007. Genetic structure and extinction of the woolly mammoth, Mammuthus primigenius. Curr Biol. 17:1072-1075; Debruyne R, Chu G, King CE, Bos K, Kuch M, Schwarz C, Szpak P, Gröcke DR, Matheus P, Zazula G, et al. 2008. Out of America: ancient DNA evidence for a new world origin of late quaternary woolly mammoths. Curr Biol. 18:1320-1326).


Stanene based gas sensors: effect of spin-orbit coupling.
Density functional theory calculations are performed to investigate the gas sensing properties (NO, NO2, NH3 and N2O) of pure and doped (B@, N@, and B-N@) stanene. Dispersion corrected (DFT-D3) density functional calculations show that doping improves the interaction between stanene and gas molecules. The extent of interaction between the system and gas molecules is further studied through charge density difference (CDD), electrostatic potential (ESP) and Bader charge analysis. The electronic properties of pure stanene + gases are studied with and without the effect of spin-orbit coupling. Stanene + gas systems show the Rashba-type of spin-splitting under spin-orbit coupling (SOC), which is very promising for spintronic applications. Interestingly, the doped systems (B@-, N@-, and B-N@stanene) show higher selectivity and sensitivity toward gas molecules compared to pure stanene. Therefore, the B@-, N@-, and B-N@stanene systems are promising for semiconductor based gas sensors.


Comparison of Cardiovascular Benefits of Bariatric Surgery and Abdominal Lipectomy.
The purpose of this review is to examine recent evidence supporting effectiveness of bariatric surgery and abdominal lipectomy as interventional strategies aimed at reduction in incidence of cardiovascular disease (CVD) and related morbidity and mortality in obese and metabolic syndrome patients. While several studies show reduction in CVD risk factors in patients who have undergone both the Roux-en-Y gastric bypass and sleeve gastrectomy, very few demonstrate actual improvements in cardiovascular function, or a decrease in CVD events or CVD-related mortality. Consequently, the cardiovascular benefits of the less invasive sleeve gastrectomy in comparison to the gastric bypass are also unclear. Striking new data on large patient samples demonstrate significant positive correlation between gastric bypass and CVD risk factor reduction only in patients who are diabetic or > 50 years of age at the time of surgery, with no significant differences in non-diabetic and younger patients and with significant side effects. On the other hand, a markedly less invasive removal of abdominal subcutaneous adipose tissue via lipectomy consistently and significantly improved CVD risk factors as well as cardiovascular function in the very few studies available. Overall, neither the potential nor the definitive cardiovascular benefits of either of the commonly used bariatric surgical or the various lipectomy procedures have been adequately explored. Future basic science and clinical studies have the opportunity to understand the mechanisms and long-term consequences of both approaches and develop personalized approaches with higher benefit to side effect ratios.


The role of co-regulation of stress in the relationship between perceived partner responsiveness and binge eating: A dyadic analysis.
The main aim of this study is to investigate whether there are relationships between perceived partner responsiveness (PPR), co-regulation of negative affect between romantic partners and binge eating. Data were collected from 148 opposite-sex romantic partners (18-61 years old) with the mean relationship duration being 8.04 years. Dyadic data were analysed through the actor-partner interdependence model framework. We proposed a model where co-regulation between partners has indirect effects on the relationship between PPR and binge eating. Results showed that there was no direct association between PPR and binge eating scores of the participants. However, significant direct associations were found regarding both actor and partner effects of PPR on co-regulation between romantic partners. In addition, there were four significant indirect effects: Women's co-regulation had an indirect effect on the link between PPR and women's binge eating scores. Similarly, women's co-regulation had also a significant indirect effect on the link between PPR and men's binge eating. These findings are the first to illustrate a relationship between PPR and binge eating. This study is the first attempt to examine binge eating in terms of co-regulation processes.


Interleukin-1 and tumour necrosis factor cause hypotension in the conscious rabbit.
1. The cardiovascular effects of intravenous injections of interleukin-1 (IL-1) and tumour necrosis factor (TNF) have been investigated in the conscious rabbit. They have been compared with the effects of bacterial lipopolysaccharide (LPS) because both IL-1 and TNF are released from macrophages by LPS. 2. IL-1, TNF and Escherichia coli J5-LPS all caused hypotension when given intravenously in a dose with low mortality. The time course of the hypotension caused by IL-1 and LPS was similar, although the maximal fall in mean blood pressure occurred earlier after IL-1. TNF produced a more sustained fall in blood pressure. Hypotension was not accompanied by a compensatory tachycardia after any of the test substances. Hypotension was associated with a fever after TNF, hypothermia after LPS and no significant change in temperature after IL-1. 3. The packed cell volume did not change during hypotension in any of the study groups, implying that the hypotension was not due to fluid loss resulting from increased capillary permeability. 4. IL-1 and TNF are candidates for the role of effectors of LPS-induced hypotension.


On-a-chip tryptic digestion of transthyretin: a step toward an integrated microfluidic system for the follow-up of familial transthyretin amyloidosis.
A microfluidic microreactor for trypsin mediated transthyretin (TTR) digestion has been developed as a step towards the elaboration of a fully integrated microdevice for the detection of a rare and disabling disease, the familial transthyretin amyloidosis (ATTR) which is related to specific TTR mutations. Therefore, an enzymatic microreactor coupled to an analytical step able to monitor the mutation of TTR on specific peptide fragments would allow an accurate monitoring of the treatment efficiency of ATTR. In this study, two types of immobilized trypsin microreactors have been investigated: a new miniaturized, microfluidic fluidized bed packed with trypsin functionalized magnetic particles (MPs), and a thiol-ene (TE) monolith-based chip. Their performances were first demonstrated with N-benzoyl-dl-arginine-4-nitroanilide hydrochloride BApNA, a low molecular weight substrate. High reaction yields (75.2%) have been reached within 0.6 min for the TE-based trypsin microreactor, while a lower yield (12.4%) was obtained for the micro-fluidized bed within a similar residence time. Transposition of the optimized conditions, developed with BApNA, to TTR digestion in the TE-based trypsin microreactor was successfully performed. We demonstrated that the TE-chip can achieve an efficient and reproducible digestion of TTR. This has been assessed by MS detection. In addition, TTR hydrolysis led to the production of a fragment of interest allowing the therapeutic follow-up of more than twenty possible ATTR mutations. High sequence coverage (90%), similar to those obtained with free trypsin, was achieved in a short time (2.4 min). Repeated experiments showed good reproducibility (RSD = 6.8%). These promising results open up the route for an innovative treatment follow-up dedicated to ATTR.


The everyday practice of supporting health system development: learning from how an externally-led intervention was implemented in Mozambique.
Health system strengthening (HSS) has often been undertaken by global health actors working through vertical programmes. However, experience has shown the challenges of this approach, and the need to recognize health systems as open complex adaptive systems-which in turn has implications for the design and implementation approach of more 'horizontal' HSS interventions. From 2009 to 2016, the Doris Duke Charitable Foundation supported the African Health Initiative, establishing Population Health Implementation and Training partnerships in five African countries (Ghana, Mozambique, Rwanda, Tanzania and Zambia). Each partnership was designed as a large-scale, long-term, complex health system strengthening intervention, at a primary care or district level-and in each country the intervention was adapted to suit that specific health systems context. In Mozambique, the Population Health Implementation and Training partnership sought to strengthen integrated health systems management at district and provincial levels (through a variety of capacity-development intervention activities, including in-service training and mentoring); to improve the quality of routine data and develop appropriate tools to facilitate decision-making for provincial and district managers; and to build capacity to design and conduct innovative operations research in order to guide integration and system-strengthening efforts. The success of this intervention, as assessed by outcome measures, has been reported elsewhere. In this paper, the implementation practice of this horizontal HSS intervention is assessed, focusing on the key features of how implementation occurred and the implementation approach. A case study focusing on HSS implementation practice was conducted by external researchers from 2014 to 2017. The importance of an accompanying implementation research approach is emphasized-especially for HSS interventions where the 'complex adaptive system' (complex and constantly changing context) forces constant adaptations to the intervention design and approach.


Treatment and Survival of Small-bowel Adenocarcinoma in the United States: A Comparison With Colon Cancer.
Small-bowel adenocarcinoma is rare and fatal. Because of data paucity, there is a tendency to extrapolate treatment from colon cancer, particularly in the adjuvant stetting. The purpose of this study was to evaluate the current surgical and adjuvant treatments of small-bowel adenocarcinoma and compare with colon cancer. This was a retrospective cohort study. The linked Surveillance, Epidemiology, and End Results and Medicare database was used at a tertiary referral hospital. Patients with small-bowel adenocarcinoma and colon cancer identified from 1992 to 2010, using International Classification of Diseases for Oncology, 3 Revision, site, behavior, and histology codes were included. Overall survival and cancer-specific survival were estimated using the Kaplan-Meier method and competing risk analysis. A total of 2123 patients with small-bowel adenocarcinoma and 248,862 patients with colon cancer were identified. Five-year overall survival rates for patients with small-bowel adenocarcinoma and colon cancer were 34.9% and 51.5% (p < 0.0001). A total of 1550 patients with small-bowel adenocarcinoma (73.0%) underwent surgery, compared with 177,017 patients with colon cancer (71.1%). The proportion of patients who received chemotherapy was similar, at 21.3% for small bowel and 20.0% for colon. In contrast to colon cancer, chemotherapy did not improve overall or cancer-specific survival for patients with small-bowel adenocarcinoma, regardless of stage. Predictors of poor survival for small-bowel adenocarcinoma on multivariate analysis included advanced age, black race, advanced stage, poor tumor differentiation, high comorbidity index, and distal location. Chemotherapy did not confer additional survival benefit compared with surgery alone (HR, 1.04 (95% CI, 0.90-1.22)). This was a retrospective review. The reliance on Medicare data limited granularity and may have affected the generalizability of the results. The prognosis for small-bowel adenocarcinoma is worse than that for colon cancer, and only surgery improves survival. In contrast to colon cancer, a survival benefit from current chemotherapy regimens for small-bowel adenocarcinoma is not seen, suggesting that it may be overused and needs more rigorous study.


Distribution of tamoxifen and its metabolites in rat and human tissues during steady-state treatment.
A procedure for the extraction of tamoxifen and metabolites from various rat and human tissues was developed and verified. With this method, we determined the drug and metabolite concentrations during one dosing interval in various tissues (brain, fat, liver, heart, lung, kidney, uterus, and testes) of rats given tamoxifen once daily for 3 or 14 days, and in various normal and malignant tissues obtained during surgery or at autopsy from patients with breast cancer treated with tamoxifen. In the rat, the concentrations of tamoxifen and metabolites in most tissues were 8- to 70-fold higher than in serum. The highest levels were observed in lung and liver; substantial amounts were also recovered from kidney and fat. Fluctuations of metabolites and tamoxifen content in most tissues were observed during one dosing interval, corresponding to a ratio of 4:8 between Cmax and Cmin, except in fat and testicular tissues, where the drug concentrations were relatively stable. In addition to tamoxifen, N-desmethyltamoxifen, followed by 4-hydroxytamoxifen, 4-hydroxy-N-desmethyltamoxifen, and N-desdimethyltamoxifen, were abundant in most tissues. In contrast, adipose tissue contained only small amounts of these metabolites. The concentrations of tamoxifen and metabolites found in human normal and malignant tissues confirmed and extended the conclusions made in the experiments with rats. In humans, levels were 10- to 60-fold higher in tissues than in serum, and relatively high concentrations were detected in liver and lung. Additionally, pancreas, pancreatic tumor, and brain metastases from breast cancer and primary breast cancer retained large amounts of drug. Again, the amounts of demethylated and hydroxylated metabolites were high in most tissues, except in fat. Tamoxifen and some metabolites were also present in specimens of skin and bone tissue. In one patient, significant amounts of drugs could be detected in lung, heart, ovary, and intestinal wall 14 months after withdrawal of tamoxifen, demonstrating efficient retention and slow washout of these compounds in human tissue.


Demographic Characteristics and Association of Serum Vitamin B12, Ferritin and Thyroid Function with Premature Canities in Indian Patients from an Urban Skin Clinic of North India: A Retrospective Analysis of 71 Cases.
The incidence of self-reported premature hair graying (PHG) seems to be on the rise. PHG has a profound impact on the patient's quality of life. It remains an incompletely understood etiology with limited and modest treatment options. The evaluation of the demographic and clinical profile of patients with premature canities, and exploration of the association of this entity with certain systemic disorders suspected to be related to its etiology. Seventy-one cases of premature canities (onset noticed by patients before 25 years of age) presenting to an urban skin clinic in Gurugram, India, between September 2012 and September 2015 with this complaint were retrospectively analyzed. The patient records were retrieved that provided details of the onset, duration and pattern of involvement, history, and examination findings (scalp, cutis, and general physical). Since all these patients had been screened for anemia, thyroid disorder, fasting blood glucose, and Vitamin B12 levels at the time of presentation, these parameters were also available for analysis. The mean age at onset of graying was 10.2 ± 3.6 years (range: 5-19 years), with an almost equal gender distribution. The earliest age of onset recorded was 5 years. A positive family history of PHG (at least one of the biological parents or siblings) was obtained in 64 (90.1%) of the cases. The temporal regions of the scalp (35.2%) were most commonly involved followed by the frontal region (18.3%). Hypovitaminosis B12 and hypothyroidism showed significant association with the disorder, whereas anemia, serum ferritin, and fasting blood glucose did not. The age of onset of hair graying can be as low as 5 years. Temporal and frontal areas are the most commonly involved sites. A strong family history, Vitamin B12 deficiency, and hypothyroidism are strongly associated with PHG. Larger case-control studies are mandated for discerning the correlation of these and other risk factors with PHG.


Fructose metabolism in humans - what isotopic tracer studies tell us.
Fructose consumption and its implications on public health are currently under study. This work reviewed the metabolic fate of dietary fructose based on isotope tracer studies in humans. The mean oxidation rate of dietary fructose was 45.0% ± 10.7 (mean ± SD) in non-exercising subjects within 3-6 hours and 45.8% ± 7.3 in exercising subjects within 2-3 hours. When fructose was ingested together with glucose, the mean oxidation rate of the mixed sugars increased to 66.0% ± 8.2 in exercising subjects. The mean conversion rate from fructose to glucose was 41% ± 10.5 (mean ± SD) in 3-6 hours after ingestion. The conversion amount from fructose to glycogen remains to be further clarified. A small percentage of ingested fructose (<1%) appears to be directly converted to plasma TG. However, hyperlipidemic effects of larger amounts of fructose consumption are observed in studies using infused labeled acetate to quantify longer term de novo lipogenesis. While the mechanisms for the hyperlipidemic effect remain controversial, energy source shifting and lipid sparing may play a role in the effect, in addition to de novo lipogenesis. Finally, approximately a quarter of ingested fructose can be converted into lactate within a few of hours. The reviewed data provides a profile of how dietary fructose is utilized in humans.


c-myb activates CXCL12 transcription in T47D and MCF7 breast cancer cells.
Chemokine C-X-C motif ligand 12 (CXCL12) is a potent chemotactic and angiogenic factor that has been proposed to play a role in organ-specific metastasis and angiogenic activity in several malignancies. In this study, we found that the overexpression of c-myb could elevate CXCL12 mRNA level and CXCL12 promoter activity in human T47D and MCF-7 breast cancer cells. Chromatin immunoprecipitation assay demonstrated that c-myb could bind to the CXCL12 promoter in the cells transfected with cmyb expression vector. c-myb siRNA attenuated CXCL12 promoter activity and the binding of c-myb to the CXCL12 promoter in T47D and MCF-7 cells. These results indicated that c-myb could activate CXCL12 promoter transcription.


Formulation of Binucleate Rhizoctonia spp. and Biocontrol of Rhizoctonia solani on Impatiens.
Isolates BNR621 and P9023 of binucleate Rhizoctonia spp. (BNR) in Pesta and rice flour formulations were evaluated for control of preemergence damping-off of impatiens caused by R. solani. Amendment of a soilless potting mix with the formulations at 0.47% (vol:vol) 3 days prior to seeding and infesting did not improve control compared to amendment 1 day prior to seeding and infesting regardless of whether the moistened amended potting mix was stored in closed plastic bags or in plug trays under a mist system. BNR fungi were no more effective in biocontrol of R. solani in formulations amended at 0.9%. Control of damping-off was comparable but not consistent between formulations of BNR fungi and the fungicide thiophanatemethyl. Damping-off was controlled better with formulations of BNR fungi than with SoilGard based on Trichoderma virens. Shelf life of Pesta and rice flour formulations at 4°C was determined by assessing viability of BNR isolates over time. Viability of the BNR isolates, measured as CFU/g of formulation, declined to approximately 68 to 79% of the original propagule concentration after 6 months in Pesta and rice flour formulations, with the greatest decline in the first 2 months. Shelf life of BNR isolates in formulation significantly affected control of preemergence damping-off but was isolate dependent. Preemergence damping-off was only 5 to 7% with fresh formulations but increased to 30 to 50% with 4-month-old formulations. Controlled atmospheres, maintained with saturated salt solutions, were established to measure the effect of water activity on shelf life of formulations. Water activities (aw) of 0.12 and 0.33 aw enhanced BNR survival in formulations by approximately 2 to 3 months compared with aw of 0.53 and 0.75 aw. Storage of Pesta and rice flour formulations at 4°C significantly improved BNR survival by 4 to 5 months compared with storage at 25°C. These results suggest that improved shelf life of BNR isolates is needed before formulated products can be developed for biocontrol of preemergence damping-off.


Chondromalacia of the patella.
One hundred and sixty-three patients who underwent arthroscopic studies of the knee have been reviewed in an attempt to better understand the clinical condition present in chondromalacia of the patella. All patients included in the study were found to have either painful knees, patellar chondromalacia, or both, but no other intraarticular pathology. The difference between true chondromalacia of the patella (which may or may not be symptomatic) and other types of pain in the patello-femoral area is emphasized. Although various types of conservative and operative treatment have been described, the symptoms have frequently been found to resolve spontaneously, especially in young patients.


A homozygous splice site mutation affecting the intracellular domain of the growth hormone (GH) receptor resulting in Laron syndrome with elevated GH-binding protein.
Laron syndrome (LS) is a severe autosomal recessive form of GH resistance resulting from molecular defects in the GH receptor (GHR). Affected individuals have extreme short stature and a typical facial phenotype. The point mutations in the GHR gene identified in this condition have until now been confined to the region encoding the extracellular domain of the receptor. We report here the first homozygous point mutation within the intracellular domain of the GHR in two LS cousins distinguishable from classical LS patients only by the presence of elevated GH-binding protein (GHBP) in their serum. A G to C transversion at the vital - 1 position in the splice donor site of exon 8 disrupts normal splicing, resulting in the complete skipping of exon 8, producing a mutant GHR protein lacking transmembrane and intracellular domains. We predict that this mutant protein would not be anchored in the cell membrane and would be measurable in the circulation as GHBP, hence explaining the phenotype of severe GH resistance combined with elevated circulating GHBP.


Tree-sequence recording in SLiM opens new horizons for forward-time simulation of whole genomes.
There is an increasing demand for evolutionary models to incorporate relatively realistic dynamics, ranging from selection at many genomic sites to complex demography, population structure, and ecological interactions. Such models can generally be implemented as individual-based forward simulations, but the large computational overhead of these models often makes simulation of whole chromosome sequences in large populations infeasible. This situation presents an important obstacle to the field that requires conceptual advances to overcome. The recently developed tree-sequence recording method (Kelleher, Thornton, Ashander, & Ralph, 2018), which stores the genealogical history of all genomes in the simulated population, could provide such an advance. This method has several benefits: (1) it allows neutral mutations to be omitted entirely from forward-time simulations and added later, thereby dramatically improving computational efficiency; (2) it allows neutral burn-in to be constructed extremely efficiently after the fact, using "recapitation"; (3) it allows direct examination and analysis of the genealogical trees along the genome; and (4) it provides a compact representation of a population's genealogy that can be analysed in Python using the msprime package. We have implemented the tree-sequence recording method in SLiM 3 (a free, open-source evolutionary simulation software package) and extended it to allow the recording of non-neutral mutations, greatly broadening the utility of this method. To demonstrate the versatility and performance of this approach, we showcase several practical applications that would have been beyond the reach of previously existing methods, opening up new horizons for the modelling and exploration of evolutionary processes.


Gliadin-specific, HLA-DQ(alpha 1*0501,beta 1*0201) restricted T cells isolated from the small intestinal mucosa of celiac disease patients.
Celiac disease (CD) is most probably an immunological disease, precipitated in susceptible individuals by ingestion of wheat gliadin and related proteins from other cereals. The disease shows a strong human HLA association predominantly to the cis or trans encoded HLA-DQ(alpha 1*0501,beta 1*0201) (DQ2) heterodimer. T cell recognition of gliadin presented by this DQ heterodimer may thus be of immunopathogenic importance in CD. We therefore challenged small intestinal biopsies from adult CD patients on a gluten-free diet in vitro with gluten (containing both gliadin and other wheat proteins), and isolated activated CD25+ T cells. Polyclonal T cell lines and a panel of T cell clones recognizing gluten were established. They recognized the gliadin moiety of gluten, but not proteins from other cereals. Inhibition studies with anti-HLA antibodies demonstrated predominant antigen presentation by HLA-DQ molecules. The main antigen-presenting molecule was established to be the CD-associated DQ(alpha 1*0501, beta 1*0201) heterodimer. The gluten-reactive T cell clones were CD4+, CD8-, and carried diverse combinations of T cell receptor (TCR) V alpha and V beta chains. The findings suggest preferential mucosal presentation of gluten-derived peptides by HLA-DQ(alpha 1*0501, beta 1*0201) in CD, which may explain the HLA association.


The method and accuracy of documentation of intraoperative fluids management in liver transplantation recipients.
In liver transplantation, blood loss can be massive, requiring timely and rapid fluid resuscitation. Maintaining proper documentation of fluids during such situations can be difficult and may often lead to counting errors. We report our method of documentation of fluid management during liver transplantation. Each unit of red blood cells (125 cc) that comes from the blood bank had a serial number of 10 Arabic numbers which were verified and double-checked. Each unit was then numbered and labeled as encircled absolute numbers (e.g., 1, 2, 3). Both the encircled number and the serial number of the bag were recorded in the anesthesia chart. Each liter of crystalloids and colloids were similarly numbered and labeled in sequence for ease of calculation. At the end of the operation, the nurse anesthetist ascertains that the number of units of blood products used matched with the number of units supplied by the blood bank. The total amounts of crystalloids and colloids given during the operation was also calculated, rechecked and written in a tabulated form. Since the introduction of this method, we have detected and readily corrected 3 incidences of counting discrepancy in the total units of blood products transfused and the products supplied by the blood bank. Moreover, our records have now become transparent data that are easily retrievable for future scientific research. Our method of documentation of fluid management during liver transplantation is easy, accurate and effective.


Consistent Video Saliency Using Local Gradient Flow Optimization and Global Refinement.
We present a novel spatiotemporal saliency detection method to estimate salient regions in videos based on the gradient flow field and energy optimization. The proposed gradient flow field incorporates two distinctive features: 1) intra-frame boundary information and 2) inter-frame motion information together for indicating the salient regions. Based on the effective utilization of both intra-frame and inter-frame information in the gradient flow field, our algorithm is robust enough to estimate the object and background in complex scenes with various motion patterns and appearances. Then, we introduce local as well as global contrast saliency measures using the foreground and background information estimated from the gradient flow field. These enhanced contrast saliency cues uniformly highlight an entire object. We further propose a new energy function to encourage the spatiotemporal consistency of the output saliency maps, which is seldom explored in previous video saliency methods. The experimental results show that the proposed algorithm outperforms state-of-the-art video saliency detection methods.