Borax-Loaded PLLA for Promotion of Myogenic Differentiation.
Boron is an essential metalloid, which plays a key role in plant and animal metabolisms. It has been reported that boron is involved in bone mineralization, has some uses in synthetic chemistry, and its potential has been only recently exploited in medicinal chemistry. However, in the area of tissue engineering, the use of boron is limited to works involving certain bioactive glasses. In this study, we engineer poly(l-lactic acid) (PLLA) substrates with sustained release of boron. Then, we analyze for the first time the uniqueness effects of boron in cell differentiation using murine C2C12 myoblasts and discuss a potential mechanism of action in cooperation with Ca(2+). Our results demonstrate that borax-loaded materials strongly enhance myotube formation at initial steps of myogenesis. Furthermore, we demonstrate that Ca(2+) plays an essential role in combination with borax as chelating or blocking Ca(2+) entry into the cell leads to a detrimental effect on myoblast differentiation observed on borax-loaded materials. This research identifies borax-loaded materials to trigger differentiation mechanisms and it establishes a new tool to engineer microenvironments with applications in regenerative medicine for muscular diseases.


The p53 codon 72 polymorphism (rs1042522) is associated with proliferative vitreoretinopathy: the Retina 4 Project.
To compare the distribution of a p53 gene polymorphism among European subjects undergoing primary retinal detachment (RD) surgery in relation to the development of proliferative vitreoretinopathy (PVR). Case-controlled gene association study conducted as a component of the Retina 4 Project (a European multicenter study). Five hundred fifty DNA samples, 134 with PVR secondary to primary RD and 416 with RD without PVR. The p53 codon 72 polymorphism (rs1042522) was analyzed using allele-specific primer polymerase chain reaction. Proportions of genotypes and the proline (Pro-P) homozygote groups between subsamples from different countries were analyzed in 2 phases. In the first, subsamples from Spain and Portugal were analyzed. After significant results were found, samples from the United Kingdom (UK) and The Netherlands were analyzed (second phase). Genotypic and allelic frequencies were compared between cases and controls in the global sample. Single significant associations with PVR. A significant difference (P<0.05, Fisher exact test) was observed regarding the p53 genotype frequencies at codon 72 between the PVR cases and the non-PVR controls in Spain and Portugal (phase I), but not in the UK or The Netherlands (phase II). Analysis of Pro homozygote carriers between cases and controls revealed differences in Spain (29.01-42.18 and 2.29-10.20, respectively), Portugal (10.49-29.50 and 1.35-8.89, respectively), and The Netherlands (16.49-31.70 and 4.51-15.09, respectively), but no differences in the UK (7.68-18.1 and 4.85-13.94, respectively). The odds ratio of Pro carriers from Spain and Portugal together was 8.12 (95% confidence interval [CI], 3.72-17.69; P<0.05), whereas the odds ratio of Pro carriers from the UK and The Netherlands was 2.12 (95% CI, 0.96-4.68; P = 0.07). All control samples were in Hardy-Weinberg equilibrium. Considering the entire sample, significant differences were found in genotype frequencies between cases (RR, 30.59%; RP, 43.28%; PP, 26.11% [R = Arg; P = Pro]) and controls (RR, 39.66%; RP, 52.64%; PP, 7.69%) and in Pro homozygote carriers between controls (Pro homozygote 95% CI, 18.67-33.52) and cases (Pro homozygote 95% CI, 5.1-10.2). Results indicate that the Pro variant of p53 codon 72 polymorphism is associated with a higher risk of PVR developing after a primary RD. Further studies are necessary to understand the role of this polymorphism in the development of PVR.


Coach Rating Combined With Small-Sided Games Provides Further Insight Into Mental Toughness in Sport.
Literature indicates that mental toughness contributes to successful performance when faced with challenge. This study used an exemplar sport of Australian Rules football to investigate whether skilled performance thrived across increased challenge in small-sided games. Higher (n = 14) and lower (n = 17) skilled Australian footballers were recruited. First, coaches rated participants' mental toughness (MTC) using the Mental Toughness Index. Second, participants competed in small-sided games where challenge was manipulated by varying the attacker to defender ratio to create lower and higher pressure scenarios. Decision-making, motor skill execution, and a combined total were measured. MTC rating was higher for higher skilled players. Total score of higher skilled players was significantly superior to lower skilled players in higher and lower pressure scenarios (p = 0.003). A "pressure differential score," calculated to determine whether participants maintained performance across increased challenge, indicated a significant decrease in performance (total score) from lower to higher pressure scenarios for lower skilled (p = 0.011), but not for higher skilled (p = 0.060) players. Furthermore, MTC scores were predictive of high pressure scenario total scores (p = 0.011). Findings suggest higher levels of mental toughness may contribute to maintain performance across the increased challenge of pressure within small-sided games. Practitioners can subjectively rate athlete mental toughness and then structure small-sided games to objectively measure performance under pressure scenarios. This provides an interdisciplinary approach to assess and train psychomotor skill.


Repetitive Sampling and Control Threshold Improve 16S rRNA Gene Sequencing Results From Produced Waters Associated With Hydraulically Fractured Shale.
Sequencing microbial DNA from deep subsurface environments is complicated by a number of issues ranging from contamination to non-reproducible results. Many samples obtained from these environments - which are of great interest due to the potential to stimulate microbial methane generation - contain low biomass. Therefore, samples from these environments are difficult to study as sequencing results can be easily impacted by contamination. In this case, the low amount of sample biomass may be effectively swamped by the contaminating DNA and generate misleading results. Additionally, performing field work in these environments can be difficult, as researchers generally have limited access to and time on site. Therefore, optimizing a sampling plan to produce the best results while collecting the greatest number of samples over a short period of time is ideal. This study aimed to recommend an adequate sampling plan for field researchers obtaining microbial biomass for 16S rRNA gene sequencing, applicable specifically to low biomass oil and gas-producing environments. Forty-nine different samples were collected by filtering specific volumes of produced water from a hydraulically fractured well producing from the Niobrara Shale. Water was collected in two different sampling events 24 h apart. Four to five samples were collected from 11 specific volumes. These samples along with eight different blanks were submitted for analysis. DNA was extracted from each sample, and quantitative polymerase chain reaction (qPCR) and 16S rRNA Illumina MiSeq gene sequencing were performed to determine relative concentrations of biomass and microbial community composition, respectively. The qPCR results varied across sampled volumes, while no discernible trend correlated contamination to volume of water filtered. This suggests that collecting a larger volume of sample may not result in larger biomass concentrations or better representation of a sampled environment. Researchers could prioritize collecting many low volume samples over few high-volume samples. Our results suggest that there also may be variability in the concentration of microbial communities present in produced waters over short (i.e., hours) time scales, which warrants further investigation. Submission of multiple blanks is also vital to determining how contamination or low biomass effects may influence a sample set collected from an unknown environment.


Molecular analysis of polymerase gamma gene and mitochondrial polymorphism in fertile and subfertile men.
CAG trinucleotide repeat length in the nuclear polymerase gamma gene (POLgamma) has been shown to be associated with men with reduced fertility. The present study investigated the frequency of CAG repeat length genotypes and three exonuclease motifs of the POLgamma in relation to the frequency of mitochondrial nucleotide substitutions. DNA from semen samples of 93 normozoospermic men and 192 non-normozoospermic men was isolated and the specific regions of the genes were amplified by polymerase chain reactions (PCR) and sequenced to identify mutations. The genotypic frequencies of pooled POLgamma CAG repeat lengths, =10/ not equal 10 heterozygotes and not equal 10/ not equal 10 homozygotes, were significantly different between normozoospermic and non-normozoospermic men (p < 0.05), with non-normozoospermic men having a slightly higher frequency of the =10/=10 genotypes. The allelic frequency for =10 is 0.79 and not equal10 is 0.21 for normozoospermic men and 0.85 and 0.15, respectively, for non-normozoospermic men (p < 0.025). There was no mutation detected in the exonuclease motifs in all the samples tested. Eighty normozoospermic and 124 non-normozoospermic semen samples were analysed for nucleotide substitutions in mitochondrial genes by PCR and sequencing. Heteroplasmic mutations were found in one azoospermic man, four asthenozoospermic men and two normozoospermic men. Only one asthenozoospermic man was heterozygous for the POLgamma genotype. Of the 17 men with non-synonymous nucleotide substitutions, 14 were homozygous for the POLgamma genotype. Non-normozoospermic men had twice as many nucleotide substitutions than normozoospermic men. However, there were no significant differences in the frequencies of nucleotide substitution and POLgamma genotypes in the two groups of men.


Evaluation of the effects of nephrotomy on renal function in clinically normal cats.
To evaluate the effects of nephrotomy on renal function in clinically normal cats. 20 specific-pathogen-free, 9- to 11-month-old female mixed-breed cats. Serum chemistry analyses, CBC determinations, urinalyses, microbiologic urine cultures, renal ultrasonography, abdominal radiography, and single-kidney and total glomerular filtration rate (GFR) determinations by use of renal scintigraphy and measurements of plasma disappearance of technetium 99m-diethylenetriaminepentaacetic acid were performed before surgery and at 3, 12, 26, 52, and 78 weeks after surgery in 10 cats that underwent unilateral nephrotomy and in 10 control cats that underwent a sham surgical procedure. Two cats (1 from each group) did not complete the study, and their data were eliminated from analyses. Unilateral nephrotomy resulted in a 10% to 20% reduction in mean single-kidney GFR, compared with that of nephrotomy contralateral control kidneys. However, mean total GFR in nephrotomy-group cats was not significantly different from that of sham-group cats. Over the 78 weeks of study, mean total GFR declined 34% and 40% in nephrotomy- and sham-group cats, respectively. Adverse events associated with nephrotomy included persistent microscopic hematuria, renal pelvis hyperechogenicity with distant shadowing on ultrasonographic evaluation, dilatation of renal pelves, and hydronephrosis. Nephrotomy in normal functioning feline kidneys results in a modest relative reduction in renal function, compared with contralateral kidney controls, but has minimal effect on total GFR when compared with sham-operated control cats. However, any detrimental effects of nephrotomy may be magnified in cats with diseased kidneys, which may have little or no capacity for repair or compensation.


Rapid, Dynamic Activation of Müller Glial Stem Cell Responses in Zebrafish.
Zebrafish neurons regenerate from Müller glia following retinal lesions. Genes and signaling pathways important for retinal regeneration in zebrafish have been described, but our understanding of how Müller glial stem cell properties are regulated is incomplete. Mammalian Müller glia possess a latent neurogenic capacity that might be enhanced in regenerative therapies to treat degenerative retinal diseases. To identify transcriptional changes associated with stem cell properties in zebrafish Müller glia, we performed a comparative transcriptome analysis from isolated cells at 8 and 16 hours following an acute photic lesion, prior to the asymmetric division that produces retinal progenitors. We report a rapid, dynamic response of zebrafish Müller glia, characterized by activation of pathways related to stress, nuclear factor-κB (NF-κB) signaling, cytokine signaling, immunity, prostaglandin metabolism, circadian rhythm, and pluripotency, and an initial repression of Wnt signaling. When we compared publicly available transcriptomes of isolated mouse Müller glia from two retinal degeneration models, we found that mouse Müller glia showed evidence of oxidative stress, variable responses associated with immune regulation, and repression of pathways associated with pluripotency, development, and proliferation. Categories of biological processes/pathways activated following photoreceptor loss in regeneration-competent zebrafish Müller glia, which distinguished them from mouse Müller glia in retinal degeneration models, included cytokine signaling (notably NF-κB), prostaglandin E2 synthesis, expression of core clock genes, and pathways/metabolic states associated with pluripotency. These regulatory mechanisms are relatively unexplored as potential mediators of stem cell properties likely to be important in Müller glial cells for successful retinal regeneration.


Role of cell-mediated immunity in tumor eradication by hyperthermia.
A single hyperthermic treatment (44.5 degrees for 20 min) which results in successful local control of a 7-day intradermally growing fibrosarcoma in intact mice fails to permanently control tumor growth in immunodeficient nu/nu (BALB/c) or immunosuppressed (whole-body radiation, 500 rads) mice. Studies were designed to determine whether humoral or cellular-mediated factors were responsible. Nude mice were reconstituted with either heterozygotic (nu/+) sensitized or nonsensitized splenic T-cells. Similarly, semisyngeneic immunosuppressed BALB/c x C57BL/6 F1 hybrids were reconstituted with homologous sensitized and nonsensitized T-cells or sera prior to hyperthermic treatment of a 7-day intradermal Meth-A implant. Successful local tumor control by hyperthermia was effected in those animals reconstituted with greater than or equal to 1.8 x 10(7) splenic T-cells but not in those with sera. The inhibition of macrophage activity (chronic silica, i.p.) could substitute for whole-body radiation immunosuppression. These studies indicate that a thermally induced tumor cure would appear to be mediated by an activated macrophage-antigen-T-cell interaction which may be dependent on the initial expression of cell-mediated antitumor immunity that may be generated only in response to immunogenic tumors.


Interfilament interaction between IMPDH and CTPS cytoophidia.
Inosine monophosphate dehydrogenase (IMPDH) and cytidine triphosphate synthase (CTPS) are two metabolic enzymes that perform rate-limiting steps in the de novo synthesis of purine and pyrimidine nucleotides, respectively. It has been shown that IMPDH and CTPS can comprise a filamentous macrostructure termed the cytoophidium, which may play a role in regulation of their catalytic activity. Although these two proteins may colocalise in the same cytoophidium, how they associate with one another is still elusive. As reported herein, we established a model HeLa cell line coexpressing OFP-tagged IMPDH2 and GFP-tagged CTPS1 and recorded the assembly, disassembly and movement of the cytoophidium in live cells. Moreover, by using super-resolution confocal imaging, we demonstrate how IMPDH- and CTPS-based filaments are aligned or intertwined in the mixed cytoophidium. Collectively, our findings provide a panorama of cytoophidium dynamics and suggest that IMPDH and CTPS cytoophidia may coordinate by interfilament interaction.


Engaging with complexity to improve the health of indigenous people: a call for the use of systems thinking to tackle health inequity.
The 400 million indigenous people worldwide represent a wealth of linguistic and cultural diversity, as well as traditional knowledge and sustainable practices that are invaluable resources for human development. However, indigenous people remain on the margins of society in high, middle and low-income countries, and they bear a disproportionate burden of poverty, disease, and mortality compared to the general population. These inequalities have persisted, and in some countries have even worsened, despite the overall improvements in health indicators in relation to the 15-year push to meet the Millennium Development Goals. As we enter the Sustainable Development Goals (SDGs) era, there is growing consensus that efforts to achieve Universal Health Coverage (UHC) and promote sustainable development should be guided by the moral imperative to improve equity. To achieve this, we need to move beyond the reductionist tendency to frame indigenous health as a problem of poor health indicators to be solved through targeted service delivery tactics and move towards holistic, integrated approaches that address the causes of inequalities both inside and outside the health sector. To meet the challenge of engaging with the conditions underlying inequalities and promoting transformational change, equity-oriented research and practice in the field of indigenous health requires: engaging power, context-adapted strategies to improve service delivery, and mobilizing networks of collective action. The application of systems thinking approaches offers a pathway for the evolution of equity-oriented research and practice in collaborative, politically informed and mutually enhancing efforts to understand and transform the systems that generate and reproduce inequities in indigenous health. These approaches hold the potential to strengthen practice through the development of more nuanced, context-sensitive strategies for redressing power imbalances, reshaping the service delivery environment and fostering the dynamics of collective action for political reform.


Individual and community resilience factors among lesbian, gay, bisexual, queer and questioning youth and adults in Israel.
Drawing on resilience theories, this study examined the individual and community factors of Israeli lesbians, gays, bisexuals, queers, and questioning (LGBQs) that contribute to positive mental health and the degree to which individual and community protective factors mitigate the adverse effect of risk factors for poor mental health. Differences in resilience factors between LGBQ youth and adults were explored. Data were collected on 890 LGBQ youth and adults. Findings emphasize the role of community-level resilience factors in the lives of LGBQs, and that these support systems differ slightly between the two age groups. Among youth, family support was both a strong predictor for well-being and a protective factor for mental distress. Although family support was found as a resilience factor among adults as well, other community-level factors (friends' support, LGBT connectedness and having steady partner) were found as protective factors for poorer mental health. These findings suggest for efforts on fostering familial support for LGBQ youth and a multi-level system that offers support at the familial, peer, relationship and community levels for both LGBQ youth and adults.


fMRI Neurofeedback Training for Increasing Anterior Cingulate Cortex Activation in Adult Attention Deficit Hyperactivity Disorder. An Exploratory Randomized, Single-Blinded Study.
Attention Deficit Hyperactivity Disorder (ADHD) is characterized by poor cognitive control/attention and hypofunctioning of the dorsal anterior cingulate cortex (dACC). In the current study, we investigated for the first time whether real-time fMRI neurofeedback (rt-fMRI) training targeted at increasing activation levels within dACC in adults with ADHD leads to a reduction of clinical symptoms and improved cognitive functioning. An exploratory randomized controlled treatment study with blinding of the participants was conducted. Participants with ADHD (n = 7 in the neurofeedback group, and n = 6 in the control group) attended four weekly MRI training sessions (60-min training time/session), during which they performed a mental calculation task at varying levels of difficulty, in order to learn how to up-regulate dACC activation. Only neurofeedback participants received continuous feedback information on actual brain activation levels within dACC. Before and after the training, ADHD symptoms and relevant cognitive functioning was assessed. Results showed that both groups achieved a significant increase in dACC activation levels over sessions. While there was no significant difference between the neurofeedback and control group in clinical outcome, neurofeedback participants showed stronger improvement on cognitive functioning. The current study demonstrates the general feasibility of the suggested rt-fMRI neurofeedback training approach as a potential novel treatment option for ADHD patients. Due to the study's small sample size, potential clinical benefits need to be further investigated in future studies. ISRCTN12390961.


Survivorship and Complications of Revision Total Hip Arthroplasty with a Mid-Modular Femoral Stem.
We retrospectively reviewed 161 revision THAs with diaphyseal fitting, mid-modular femoral components performed by ten surgeons at two academic medical centers. The average follow-up was 6.1 years. At final follow-up, 4 patients required re-revision for failure of the femoral component; 3 (2%) for aseptic loosening and 1 for mechanical failure of stem in setting of periprosthetic fracture. There were a total of 24 (14.9%) revisions for any reason, with the most common reason being septic failure (10 of 24). To our knowledge, this is the largest reported series of mid-term survivorship and complications of revision THA with mid-modular femoral components. Our results show that these stems have a low rate of aseptic loosening, subsidence, and mechanical failure.


Catalysis by dihydrofolate reductase and other enzymes arises from electrostatic preorganization, not conformational motions.
The proposal that enzymatic catalysis is due to conformational fluctuations has been previously promoted by means of indirect considerations. However, recent works have focused on cases where the relevant motions have components toward distinct conformational regions, whose population could be manipulated by mutations. In particular, a recent work has claimed to provide direct experimental evidence for a dynamical contribution to catalysis in dihydrofolate reductase, where blocking a relevant conformational coordinate was related to the suppression of the motion toward the occluded conformation. The present work utilizes computer simulations to elucidate the true molecular basis for the experimentally observed effect. We start by reproducing the trend in the measured change in catalysis upon mutations (which was assumed to arise as a result of a "dynamical knockout" caused by the mutations). This analysis is performed by calculating the change in the corresponding activation barriers without the need to invoke dynamical effects. We then generate the catalytic landscape of the enzyme and demonstrate that motions in the conformational space do not help drive catalysis. We also discuss the role of flexibility and conformational dynamics in catalysis, once again demonstrating that their role is negligible and that the largest contribution to catalysis arises from electrostatic preorganization. Finally, we point out that the changes in the reaction potential surface modify the reorganization free energy (which includes entropic effects), and such changes in the surface also alter the corresponding motion. However, this motion is never the reason for catalysis, but rather simply a reflection of the shape of the reaction potential surface.


Efficacy of directly-observed chloroquine-primaquine treatment for uncomplicated acute Plasmodium vivax malaria in northeast Myanmar: A prospective open-label efficacy trial.
Chloroquine (CQ) and primaquine (PQ) remain the frontline drugs for radical cure of uncomplicated P. vivax malaria in the Greater Mekong Sub-region (GMS). Recent reports of decreased susceptibility of P. vivax to CQ in many parts of the GMS raise concerns. From April 2014 to September 2016, 281 patients with uncomplicated P. vivax infection attending clinics in border settlements for internally displaced people in northeast Myanmar were recruited into this study. Patients were treated with standard regimen of 3-day CQ and concurrent 14-day PQ (3.5 mg/kg total dose) as directly observed therapy, and followed for recurrent parasitemia within 28 days post-patency. Within the 28-day follow-up period, seven patients developed recurrent parasitemia, resulting in a cumulative rate of parasite recurrence of 2.6%. Five of the seven parasitemias recurred within two weeks, and two of those failed to clear within seven days, indicating high-grade resistance. Although failure of CQ/PQ treatment of P. vivax was relatively infrequent in northeast Myanmar, this study nonetheless confirms that CQ/PQ-resistant strains do circulate in this area, some of them of a highly resistant phenotype. It is thus recommended that patients who acquire vivax malaria in Myanmar be treated an artemisinin-combination therapy along with hypnozoitocidal primaquine therapy to achieve radical cure.


Conceptualizing the role of sediment in sustaining ecosystem services: Sediment-ecosystem regional assessment (SEcoRA).
There is a growing trend to include a consideration of ecosystem services, the benefits that people obtain from ecosystems, within decision frameworks. Not more than a decade ago, sediment management efforts were largely site-specific and held little attention except in terms of managing contaminant inputs and addressing sediments as a nuisance at commercial ports and harbors. Sediments figure extensively in the Millennium Ecosystem Assessment; however, contaminated sediment is not the dominant concern. Rather, the focus is on land and water use and management on the landscape scale, which can profoundly affect soil and sediment quality, quantity and fate. Habitat change and loss, due to changes in sediment inputs, whether reductions (resulting in the loss of beaches, storm protection, nutrient inputs, etc.) or increases (resulting in lake, reservoir and wetland infilling, coral reef smothering, etc.); eutrophication and reductions in nutrient inputs, and disturbance due to development and fishing practices are considered major drivers, with significant consequences for biodiversity and the provision and resilience of ecosystem functions and services. As a mobile connecting medium between various parts of the ecosystem via the hydrocycle, sediments both contaminated and uncontaminated, play both positive and negative roles in the viability and sustainability of social, economic, and ecological objectives. How these roles are interpreted depends upon whether sediment status (defined in terms of sediment quality, quantity, location and transport) is appropriate to the needs of a given endpoint; understanding and managing the dynamic interactions of sediment status on a diverse range of endpoints at the landscape or watershed scale should be the focus of sediment management. This paper seeks to provide a language and conceptual framework upon which sediment-ecosystem regional assessments (SEcoRAs) can be developed in support of that goal.


Design of Signal Generators Using Active Elements Developed in I3T25 CMOS Technology Single IC Package for Illuminance to Frequency Conversion.
This paper presents a compact and simple design of adjustable triangular and square wave functional generators employing fundamental cells fabricated on a single integrated circuit (IC) package. Two solutions have electronically tunable repeating frequency. The linear adjustability of repeating frequency was verified in the range between 17 and 264 kHz. The main benefits of the proposed generator are the follows: A simple adjustment of the repeating frequency by DC bias current, Schmitt trigger (threshold voltages) setting by DC driving voltage, and output levels in hundreds of mV when the complementary metal-oxide semiconductor (CMOS) process with limited supply voltage levels is used. These generators are suitable to provide a simple conversion of illuminance to frequency of oscillation that can be employed for illuminance measurement and sensing in the agriculture applications. Experimental measurements proved that the proposed concept is usable for sensing of illuminance in the range from 1 up to 500 lx. The change of illuminance within this range causes driving of bias current between 21 and 52 μA that adjusts repeating frequency between 70 and 154 kHz with an error up to 10% between the expected and real cases.


Early dexamethasone treatment induces placental apoptosis in sheep.
Glucocorticoid treatment given in late pregnancy in sheep resulted in altered placental development and function. An imbalance of placental survival and apoptotic factors resulting in an increased rate of apoptosis may be involved. We have now investigated the effects of dexamethasone (DEX) in early pregnancy on binucleate cells (BNCs), placental apoptosis, and fetal sex as a determinant of these responses. Pregnant ewes carrying singleton fetuses (n = 105) were randomized to control (n = 56, 2 mL saline/ewe) or DEX treatment (n = 49, intramuscular injections of 0.14 mg/kg ewe weight per 12 hours over 48 hours) at 40 to 41 days of gestation (dG). Placentomes were collected at 50, 100, 125, and 140 dG. At 100 dG, DEX in females reduced BNC numbers, placental antiapoptotic (proliferating cell nuclear antigen), and increased proapoptotic factors (Bax, p53), associated with a temporarily decrease in fetal growth. At 125 dG, BNC numbers and apoptotic markers were restored to normal. In males, ovine placental lactogen-protein levels after DEX were increased at 50 dG, but at 100 and 140 dG significantly decreased compared to controls. In contrast to females, these changes were independent of altered BNC numbers or apoptotic markers. Early DEX was associated with sex-specific, transient alterations in BNC numbers, which may contribute to changes in placental and fetal development. Furthermore, in females, altered placental apoptosis markers may be involved.


Glutathione-associated enzymes in the human cell lines of the National Cancer Institute Drug Screening Program.
The steady state expression of glutathione S-transferases (GSTs) at both the protein and mRNA level is reported for the 60 tumor cell lines that are used for the National Cancer Institute Drug Screening Program. Individual GST isozymes were separated, identified, and quantified (with reverse-phase calibration curves) through a novel high performance liquid chromatographic procedure. GSTP1 was the predominant isozyme and was found at quantifiable levels in all but two of the cell lines. This isozyme ranged from 0.03% to 2.7% of the total cytosolic protein. For the mu family, 90% of the lines had GSTM2, 68% had GSTM3, but only 28% were positive for the M1 phenotype. The M1 proportion is lower than would be expected from the standard M1 null phenotype for human populations. Isozymes of the alpha family were detected only at very low levels in 35% of the lines. Significant quantitative correlations among enzyme activity, total enzyme protein, and mRNA were shown for GSTP1. However, such relationships were not apparent for the mu or alpha families. Levels of glutathione (GSH), and the transcript levels of other enzymes involved in GSH homeostasis were determined. gamma-Glutamyl cysteine synthetase (gamma-GCS) was present in all cell lines, but did not correlate with levels of intracellular GSH. Glyoxalase-I and gamma-glutamyl transpeptidase, both involved in GSH salvage, were found in 100% and 70% of the cell lines, respectively. Using a pattern-matching computer program, COMPARE, we compared and correlated the arrays of mRNA and protein levels with the pattern of chemosensitivity or chemoresistance of the 60 cell lines with 175 agents constituting a standard agent database. This database is composed of compounds to which a putative mechanism of action has been assigned. Although Pearson correlation coefficients relating the target and drug patterns were generally modest, when the patterns for the enzyme protein and mRNA levels for GST pi were correlated to drug sensitivity patterns, the list of 30 agents most closely matching (for which P < 0.05) was enriched with alkylating agents. gamma-GCS also showed an enrichment of alkylating agents in the COMPARE correlations, indicating that high levels of gamma-GCS may be an important determinant of resistance. In contrast, none of the other enzymes or GSH had patterns of expression that resulted in an obvious correlation to the sensitivity or resistance of alkylating agents.


Master plans for pedestrian and bicycle transportation: community characteristics.
Recent research demonstrates the importance of targeting the built environment to support individual physical activity, particularly for people experiencing health disparities. Master plans to promote biking and/or pedestrians (BPMPs) are a potential method for environmental change. This descriptive study aims to provide a snapshot of plan attributes and better understand demographic, social and transportation characteristics of communities with BPMPs. We collected a census sample of BPMPs from 4 states. Population and commuting data were obtained from national statistics. 294 master plans were included, with most plans representing municipalities. 62% of plans targeted biking only, one-fifth targeted biking and walking, and 15% targeted walking only. The sampled locations have a similar demographic profile as the overall U.S. for median age and household income, people of color, high school education, and income inequality. The degree of racial diversity of sampled communities is slightly less than the U.S. average and the percentage of people who walk to work were slightly higher. Given that communities with master plans have a similar profile as the overall U.S., BPMPs could feasibly be spread to communities throughout the country. Further research is planned to describe BPMPs in detail toward informing future plan development.


Oligonucleotide-directed construction of mutations: a gapped duplex DNA procedure without enzymatic reactions in vitro.
The gapped duplex DNA approach to oligonucleotide-directed construction of mutations (Kramer et al. 1984, Nucl. Acids Res. 12, 9441-9456) has been developed further. A procedure is described that makes in vitro DNA polymerase/DNA ligase reactions dispensable. Direct transfection of host bacteria with gdDNA molecules of recombinant phage M13 plus mutagenic oligonucleotide results in marker yields in excess of 50% (gap size 1640 nucleotides). An important feature incorporated into the mutagenic oligonucleotide is the presence of one or two internucleotidic phosphorothioate linkages immediately adjacent to the 5'-terminus. Automated preparation and biochemical properties of such compounds are described as well as their performance in oligonucleotide-directed mutagenesis. A systematic study of the following parameters influencing marker yield is reported: Gap size, length of oligonucleotide, chemical nature of oligonucleotide termini and heatshock temperature during transformation.


Detection of Borrelia burgdorferi DNA in tissues from dogs with presumptive Lyme borreliosis.
To develop a quantitative PCR assay for detection of Borrelia burgdorferi DNA in formalin-fixed, paraffin-embedded tissues; compare results of this assay with results of immunohistochemical staining of tissues from seropositive dogs; and determine whether B burgdorferi DNA could be detected in renal tissues from dogs with presumptive Lyme nephritis. Cohort study. Archived tissue samples from 58 dogs. A quantitative PCR assay was performed on formalin-fixed, paraffin-embedded tissue sections from the dogs. Results were compared with results of immunohistochemical staining, B burgdorferi serostatus, clinical signs, and necropsy findings. 38 dogs were classified as having positive or equivocal results for Lyme borreliosis, and 20 were classified as having negative results on the basis of clinical signs, serologic findings, and pathologic abnormalities. Borrelia burgdorferi DNA was amplified from tissue samples from only 4 (7%) dogs, all of which had been classified as having positive or equivocal results for Lyme borreliosis and had signs of presumptive Lyme nephritis. Results of PCR assays of renal tissue were positive for only 1 dog, and there was no agreement between results of immunohistochemical staining (ie, detection of B burgdorferi antigen) and results of the PCR assay (ie, detection of B burgdorferi DNA) for renal tissues. Results indicated that detection of B burgdorferi DNA in formalin-fixed, paraffin-embedded tissues is feasible, but that intact B burgdorferi DNA is rarely found in tissues from naturally infected dogs, even tissues from dogs with presumptive Lyme borreliosis. Further, findings support the contention that Lyme nephritis may be a sterile, immune complex disease.


Phosphorus and water recovery by a novel osmotic membrane bioreactor-reverse osmosis system.
An osmotic membrane bioreactor-reverse osmosis (OMBR-RO) hybrid system integrated with periodic microfiltration (MF) extraction was evaluated for simultaneous phosphorus and clean water recovery from raw sewage. In this hybrid system, the forward osmosis membrane effectively retained inorganic salts and phosphate in the bioreactor, while the MF membrane periodically bled them out for phosphorus recovery with pH adjustment. The RO process was used for draw solute recovery and clean water production. Results show that phosphorus recuperation from the MF permeate was most effective when the solution pH was adjusted to 10, whereby the recovered precipitate contained 15-20% (wt/wt) of phosphorus. Periodic MF extraction also limited salinity build-up in the bioreactor, resulting in a stable biological performance and an increase in water flux during OMBR operation. Despite the build-up of organic matter and ammonia in the draw solution, OMBR-RO allowed for the recovery of high quality reused water.


Information giving and involvement in treatment decisions: is more really better? Psychological effects and relation with adherence.
The aim of this study is to improve our understanding of the ways in which a medical consultation style relates to satisfaction and adherence. Participants completed questionnaires about preferred and perceived consultation styles; psychological variables such as satisfaction, anxiety and depression; and questions about taking medication. Questionnaires were completed prior to the consultation, immediately after, three months later and one year later. Doctors also completed a scale on completion of the consultation. The decisions in question related to women seeking advice about Hormone Replacement Therapy (HRT) and decisions about pain management. Patients expressed preferences for information about the treatment options and for active involvement in the decision process. The extent to which patients perceived themselves as having received information about the treatment options was a particularly consistent predictor of satisfaction. The extent to which patients perceived themselves as having been prepared for the side effects of HRT was a consistent predictor of anxiety. Perceptions of the amount of information received about medical options, including information preparing individuals for potential side effects of medication, appears to be more important to satisfaction and anxiety, than actual involvement in the decision.


Improvement in clinical outcomes after dry needling in a patient with occipital neuralgia.
The primary purpose of this case report is to outline the diagnosis, intervention and clinical outcome of a patient presenting with occipital neuralgia. Upon initial presentation, the patient described a four-year history of stabbing neck pain and headaches. After providing informed consent, the patient underwent a total of four dry needling (DN) sessions over a two-week duration. During each of the treatment sessions, needles were inserted into the trapezii and suboccipital muscles. Post-intervention, the patient reported a 32-point change in her neck disability index score along with a 28-point change in her headache disability index score. Thus, it appears that subsequent four sessions of DN over two weeks, our patient experienced meaningful improvement in her neck pain and headaches. To the best of our knowledge, this is the first case report describing DN to successfully improve clinical outcomes in a patient diagnosed with occipital neuralgia.


The effects of stabilizing and directional selection on phenotypic and genotypic variation in a population of RNA enzymes.
The distribution of variation in a quantitative trait and its underlying distribution of genotypic diversity can both be shaped by stabilizing and directional selection. Understanding either distribution is important, because it determines a population's response to natural selection. Unfortunately, existing theory makes conflicting predictions about how selection shapes these distributions, and very little pertinent experimental evidence exists. Here we study a simple genetic system, an evolving RNA enzyme (ribozyme) in which a combination of high throughput genotyping and measurement of a biochemical phenotype allow us to address this question. We show that directional selection, compared to stabilizing selection, increases the genotypic diversity of an evolving ribozyme population. In contrast, it leaves the variance in the phenotypic trait unchanged.


Mission, Organization, and Future Direction of the Serological Sciences Network for COVID-19 (SeroNet) Epidemiologic Cohort Studies.
Global efforts are needed to elucidate the epidemiology of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the underlying cause of coronavirus disease 2019 (COVID-19), including seroprevalence, risk factors, and long-term sequelae, as well as immune responses after vaccination across populations and the social dimensions of prevention and treatment strategies. In the United States, the National Cancer Institute in partnership with the National Institute of Allergy and Infectious Diseases, established the SARS-CoV-2 Serological Sciences Network (SeroNet) as the nation's largest coordinated effort to study coronavirus disease 2019. The network comprises multidisciplinary researchers bridging gaps and fostering collaborations among immunologists, epidemiologists, virologists, clinicians and clinical laboratories, social and behavioral scientists, policymakers, data scientists, and community members. In total, 49 institutions form the SeroNet consortium to study individuals with cancer, autoimmune disease, inflammatory bowel diseases, cardiovascular diseases, human immunodeficiency virus, transplant recipients, as well as otherwise healthy pregnant women, children, college students, and high-risk occupational workers (including healthcare workers and first responders). Several studies focus on underrepresented populations, including ethnic minorities and rural communities. To support integrative data analyses across SeroNet studies, efforts are underway to define common data elements for standardized serology measurements, cellular and molecular assays, self-reported data, treatment, and clinical outcomes. In this paper, we discuss the overarching framework for SeroNet epidemiology studies, critical research questions under investigation, and data accessibility for the worldwide scientific community. Lessons learned will help inform preparedness and responsiveness to future emerging diseases.


Body image dissatisfaction and eating-related psychopathology in trans individuals: a matched control study.
High levels of body dissatisfaction have already been reported in the trans population; however, the root of this dissatisfaction, and its association with eating disordered behaviours, has not been studied in-depth. This study aims to assess eating disorder risk by comparing 200 trans people, 200 people with eating disorders and 200 control participants' scores on three subscales of the Eating Disorders Inventory-2 (EDI-2) and to further explore dissatisfaction in the trans participants using the Hamburg Body Drawing Scale (HBDS). The results showed that overall participants with eating disorders scored higher than trans or control groups on all EDI-2 measures, but that trans individuals had greater body dissatisfaction than control participants and, importantly, trans males had comparable body dissatisfaction scores to eating disordered males. Drive for thinness was greater in females (cis and trans) compared with males. In relation to HBDS body dissatisfaction, both trans males and trans females reported greatest dissatisfaction not only for gender-identifying body parts but also for body shape and weight. Overall, trans males may be at particular risk for eating disordered psychopathology and other body image-related behaviours.


Low Interrater Reliability of Videourodynamic Diagnosis of Detrusor External Sphincter Dyssynergia.
Complex bladder dysfunction requires urodynamic testing, often under fluoroscopy termed videourodynamic study (VUDS), to assist the diagnosis and management. Videourodynamic study is an objective tool with high interrater reliability (IRR) for identifying detrusor overactivity. However, IRR has not been validated with disorders associated with neurogenic bladder (NGB). We aim to investigate the IRR of VUDS to diagnose detrusor external sphincter dyssynergia (DESD) among NGB patients and hypothesize a high IRR to DESD diagnosis on VUDS. Videourodynamics tracings with fluoroscopic images were rated either positive or negative for evidence of DESD by 4 raters (2 neurourologists, neurourology fellow, and urology postgraduate year 2 resident), in patients who underwent VUDS from 2013 to 2017. The study population had known NGB without previous bladder reconstruction. The IRR was determined using percent agreement and κ values. The experts had a percent agreement of 82.1% (κ = 0.26). Expert 1 and expert 2 had a percent agreement of 63.6% (κ = 0.1497) and 68.9% (κ = 0.2967), respectively, when compared with the fellow. Sensitivity and specificity ranged from 28% to 75% and 64% to 93%, respectively. The negative predictive values ranged from 90% to 95%. The IRR to diagnose DESD on VUDS was much lower than expected, even among experts, and was likely multifactorial and partially owing to lack of clinical context and lack of standardized VUDS interpretation of the electromyogram. A high negative predictive value was found among all participants. Further research is needed to evaluate factors contributing to the low reproducibility of DESD diagnosis on VUDS.


Propranolol as the first-line therapy for infantile hemangiomas: preliminary results of two centers.
Despite a mostly self-limiting course, infantile hemangiomas can cause severe functional and/or cosmetic problems. The aim of this study was to determine the efficiency of propranolol treatment on infantile hemangiomas. Sixty-seven infantile hemangioma patients were included in propranolol protocol in two institutions from 2009 to 2011. Participants included 36 boys and 31 girls. An associate protocol with radiology and pediatric cardiology was constructed for appropriate patient selection. Patients received a dose of 2 mg/kg/day, and all were admitted for the first 24 hours of therapy. Sixty-seven patients were included in the study. Mean age at the initiation of therapy was 7 months (1 to 24 months), and eleven patients were older than 12 months of age when propranolol was started. All patients showed improvement with varying responses. No side effects were detected during the treatment. Previously defined treatments for hemangiomas were efficient, yet had a limited usage because of side effects. Propranolol, with a high efficacy (not as total involution but stabilization and regression) and feasibility deserves to be the first line therapy for infantile hemangiomas even after the proliferation phase.


Assessing the availability of LLINs for continuous distribution through routine antenatal care and the Expanded Programme on Immunizations in sub-Saharan Africa.
In addition to mass distribution campaigns, the World Health Organization (WHO) recommends the continuous distribution of long-lasting insecticidal nets (LLINs) to all pregnant women attending antenatal care (ANC) and all infants attending the Expanded Programme on Immunization (EPI) services in countries implementing mosquito nets for malaria control. Countries report LLIN distribution data to the WHO annually. For this analysis, these data were used to assess policy and practice in implementing these recommendations and to compare the numbers of LLINs available through ANC and EPI services with the numbers of women and children attending these services. For each reporting country in sub-Saharan Africa, the presence of a reported policy for LLIN distribution through ANC and EPI was reviewed. Prior to inclusion in the analysis the completeness of data was assessed in terms of the numbers of LLINs distributed through all channels (campaigns, EPI, ANC, other). For each country with adequate data, the numbers of LLINs reportedly distributed by national programmes to ANC was compared to the number of women reportedly attending ANC at least once; the ratio between these two numbers was used as an indicator of LLIN availability at ANC services. The same calculations were repeated for LLINs distributed through EPI to produce the corresponding LLIN availability through this distribution channel. Among 48 malaria-endemic countries in Africa, 33 malaria programmes reported adopting policies of ANC-based continuous distribution of LLINs, and 25 reported adopting policies of EPI-based distribution. Over a 3-year period through 2012, distribution through ANC accounted for 9 % of LLINs distributed, and LLINs distributed through EPI accounted for 4 %. The LLIN availability ratios achieved were 55 % through ANC and 34 % through EPI. For 38 country programmes reporting on LLIN distribution, data to calculate LLIN availability through ANC and EPI was available for 17 and 16, respectively. These continuous LLIN distribution channels appear to be under-utilized, especially EPI-based distribution. However, quality data from more countries are needed for consistent and reliable programme performance monitoring. A greater focus on routine data collection, monitoring and reporting on LLINs distributed through both ANC and EPI can provide insight into both strengths and weaknesses of continuous distribution, and improve the effectiveness of these delivery channels.


[Counseling for physical activity as a health education strategy].
This study analyzes the prevalence of health counseling related to physical activity among users of primary health care, and explores the associated variables. This cross-sectional study included a random sample of 4,060 adults and 4,003 older adults living in areas covered by primary health care in 41 cities from seven States of South and Northeast Brazil. Prevalence of counseling for physical activity was 28.9% (95%CI: 27.3-30.4) for adults and 38.9% (95%CI: 37.3-40.6) for elders. Prevalence was consistently higher among older adults, in the Northeast, and in areas covered by the Family Health Program. Gender, socioeconomic status, smoking, sedentary lifestyle, self-reported chronic non-communicable diseases, and continuous-use medication were also associated with the outcome. Counseling for physical activity at community health centers is under-promoted, considering individual needs for healthy living. There is a need to improve participation by health professionals from different fields to promote physical activity and other healthy behaviors.


Biochemical responses of juvenile and adult Japanese quails to cyanobacterial biomass.
The aim of the present study was to evaluate differences between juvenile and adult Japanese quails in responses to the exposure to cyanobacterial biomass in the diet. The OECD 205 Guideline on Avian Dietary Toxicity Test (1984) was employed in the experiment. A total of 75 freshly hatched chicks and 30 adults were exposed to cyanobacterial biomass for 15 days and blood sampled daily and on days 5, 10 and 15, respectively. Japanese quail chicks and adults received the same daily dose of approximately 224.4 ng microcystins per gram of body weight. Biochemical responses were compared against controls. No Japanese quail chicks and adults died during the acute 15-day-cyanobacterial-biomass exposure. Biochemical responses to the biomass in diet were first observed from day 5 post exposure to cyanobacterial biomass both in chicks and adults and there were age-related differences in the parameters changed. The responses of adult birds included an increase in lactate dehydrogenase, a drop in glucose and the total antioxidant capacity as well as a 15 to 20 % inhibition of acetylcholinesterase activity. Japanese quail chicks exposed to cyanobacterial biomass for the first 15 days after hatching reacted by having hypoproteinaemia, increased concentrations of triglycerides, uric acid and the total antioxidant capacity and a drop in high-density lipoprotein cholesterol in the blood. Chicks were not found to be more susceptible to the effects of biomass exposure. It seems that, due to their physiological preparation for the oxidative stress associated with hatching, Japanese quail chicks were even better able to cope with the cyanobacterial-biomass-induced oxidative stress than adults.


Monitoring of selected veterinary antibiotics in environmental compartments near a composting facility in Gangwon Province, Korea.
Many studies have been recently reported that veterinary antibiotics released into the environment have a detrimental effect on humans such as the occurrence of antibiotic-resistant bacteria. However, only limited information is available regarding to the release of antibiotics in environmental compartments in Korea. Objectives of this study were to evaluate the concentrations of antibiotics in water, sediment, and soil adjacent to a composting facility in Korea and to determine the dilution effects of antibiotics when released into the environment. Seven antibiotics of chlortetracycline, oxytetracycline, tetracycline, sulfamethazine, sulfamethoxazole, sulfathiazole, and tylosin were evaluated by high-performance liquid chromatography-tandem mass spectrometry following pretreatment using solid-phase extraction to clean the samples. Results showed that the highest concentration of each antibiotic in both aqueous and solid samples was detected from a site adjacent to the composting facility. We also found that the studied water, sediment, and soil samples are contaminated by veterinary antibiotics throughout comparison with studies from other countries. However, relatively lower concentrations of each antibiotic were observed from the rice paddy soil located at the bottom of the water stream. Further research is necessary to continuously monitor the antibiotics release into ecosystems, thereby developing an environmental risk assessment.


Telemetered EEG-EOG during psychotic behaviors of schizophrenia.
In an effort to establish correlations between abnormal behaviors characteristic of schizophrenia and simultaneous cerebral electrical activity, EEGs and electro-oculograms (EOGs) were continuously recorded for 2 to 24 hours by radiotelemetry from 40 patients with schizophrenia and 12 normal control subjects. Trained observers recorded specific behavior patterns permitting visual and computer analysis of EEG during hallucinations, stereotypy, catatonia, psychomotor blocking, and other characteristic manifestations of schizophrenia. Electroencephalographic abnormalities consisting of focal slow or spike activity over either temporal region were found in nearly half of the patients so recorded. In contrast to the EEG during ictal episodes of epilepsy, the abnormal wave forms of schizophrenic patients seldom coincided with episodes of blocking, stereotypy, or other abnormal behaviors. Increased extraocular activity or blinking were recorded in a majority of patients, but were not consistently associated with the abnormal behavior or perceptual events.


Potential involvement of the IL-33-ST2 axis in the pathogenesis of primary Sjogren's syndrome.
To investigate the role of the interleukin (IL)-33-ST2 axis in the pathophysiology of primary Sjögren's syndrome (pSS). Serum levels of IL-33 and sST2 were determined by ELISA. The expression of IL-33 and ST2 was investigated in salivary glands (SG) by immunohistochemistry. PBMC were isolated and stimulated with IL-33, IL-12 and IL-23 and the cytokine profile response was examined by flow cytometry. Intracellular cytokine detection of IFNγ and IL-17 was performed by flow cytometry. Serum IL-33 and sST2 levels were increased in pSS patients compared with controls and patients with systemic lupus erythematosus. Expression of IL-33 was upregulated in SG with Chisholm scores of 2 and 3 of pSS patients but comparable with controls for SG with Chisholm score of 4. ST2 expression in SG was downregulated in pSS patients. IL-33 at different concentrations did not increase the secretion of pro-inflammatory cytokines but acted synergistically with IL-12 and IL-23 to promote IFNγ production. NK and NKT cells were identified as main producers of IFNγ in vitro and were found in SG of pSS patients. IL-33 is released in pSS, and acts with IL-12 and IL-23 to favour the secretion of IFNγ by NK and NKT cells.


Treatment of disseminated germ-cell tumors with cisplatin, bleomycin, and either vinblastine or etoposide.
Standard chemotherapy for disseminated germ-cell tumors includes a combination of cisplatin, vinblastine, and bleomycin, but this regimen produces substantial neuromuscular toxicity. In a randomized clinical trial in 261 men with disseminated germ-cell tumors, we substituted etoposide for the vinblastine in this regimen in half the patients to compare the efficacy and toxicity of the two treatments. Among 244 patients who could be evaluated for a response, 74 percent of those receiving the regimen including vinblastine and 83 percent of those receiving the regimen including etoposide became disease-free with or without subsequent surgery (P not significant). Among the 157 patients with high tumor volume, 61 percent became disease-free on the regimen that included vinblastine, as compared with 77 percent on the regimen that included etoposide (P less than 0.05). Survival among the patients who received etoposide was higher (P = 0.048). The regimens were similar in terms of myelosuppressive effects and pulmonary toxicity. However, the etoposide regimen caused substantially fewer paresthesias (P = 0.02), abdominal cramps (P = 0.0008), and myalgias (P = 0.00002). We conclude that etoposide with cisplatin and bleomycin is superior to vinblastine with cisplatin and bleomycin in the treatment of disseminated germ-cell tumors because of diminished neuromuscular toxicity and, among patients with advanced disease, better efficacy.


Stress hormones mediate predator-induced phenotypic plasticity in amphibian tadpoles.
Amphibian tadpoles display extensive anti-predator phenotypic plasticity, reducing locomotory activity and, with chronic predator exposure, developing relatively smaller trunks and larger tails. In many vertebrates, predator exposure alters activity of the neuroendocrine stress axis. We investigated predator-induced effects on stress hormone production and the mechanistic link to anti-predator defences in Rana sylvatica tadpoles. Whole-body corticosterone (CORT) content was positively correlated with predator biomass in natural ponds. Exposure to caged predators in mesocosms caused a reduction in CORT by 4 hours, but increased CORT after 4 days. Tadpoles chronically exposed to exogenous CORT developed larger tails relative to their trunks, matching morphological changes induced by predator chemical cue; this predator effect was blocked by the corticosteroid biosynthesis inhibitor metyrapone. Tadpole tail explants treated in vitro with CORT increased tissue weight, suggesting that CORT acts directly on the tail. Short-term treatment of tadpoles with CORT increased predation mortality, likely due to increased locomotory activity. However, long-term CORT treatment enhanced survivorship, likely due to induced morphology. Our findings support the hypothesis that tadpole physiological and behavioural/morphological responses to predation are causally interrelated. Tadpoles initially suppress CORT and behaviour to avoid capture, but increase CORT with longer exposure, inducing adaptive phenotypic changes.


What Is Shaping RT and Accuracy Distributions? Active and Selective Response Inhibition Causes the Negative Compatibility Effect.
Inhibitory control such as active selective response inhibition is currently a major topic in cognitive neuroscience. Here we analyze the shape of behavioral RT and accuracy distributions in a visual masked priming paradigm. We employ discrete time hazard functions of response occurrence and conditional accuracy functions to study what causes the negative compatibility effect (NCE)-faster responses and less errors in inconsistent than in consistent prime target conditions-during the time course of a trial. Experiment 1 compares different mask types to find out whether response-relevant mask features are necessary for the NCE. After ruling out this explanation, Experiment 2 manipulates prime mask and mask target intervals to find out whether the NCE is time-locked to the prime or to the mask. We find that (a) response conflicts in inconsistent prime target conditions are locked to target onset, (b) positive priming effects are locked to prime onset whereas the NCE is locked to mask onset, (c) active response inhibition is selective for the primed response, and (d) the type of mask has only modulating effects. We conclude that the NCE is neither caused by automatic self-inhibition of the primed response due to backward masking nor by updating response-relevant features of the mask, but by active mask-triggered selective inhibition of the primed response. We discuss our results in light of a recent computational model of the role of the BG in response gating and executive control.


Hemodynamic monitoring in Nigerian patients undergoing high-risk surgery.
Hemodynamic monitoring (HM) and optimization of cardiac output and parameters of dynamic fluid responsiveness is said to improve perioperative outcome in high-risk surgical patients (HRSP). There is insufficient data to determine the burden of care and HM practices in HRSP in Nigeria. Hence, the need to assess and document the current hemodynamic management practices of anesthetists in Nigeria regarding patients undergoing high-risk surgery. An electronic mail (E-mail) based survey was conducted among 180 consultant members of the Nigeria Society of Anaesthetists. The survey contained 24 questions that range from practice location, experience in the perioperative management of high-risk patients, expectations of care, to what is available to the anesthetists to provide such care. The survey was on for 3 months. A total of 157 E-mail messages were delivered, and 73 responses were received, giving a response rate of 46.5%. The survey showed that 67 (91.8%) of respondents provide or directly supervise anesthesia for HRSP, 50 (84%) of them do this 1-5 times a week. Noninvasive blood pressure (83.6%) was routinely monitored while the central venous pressure (CVP 35.6%), invasive blood pressure (28.8%), and cardiac output (1.4%) monitored less often. Urine output, arterial blood pressure, pulse rate, and clinical experience were considered best indicators of volume expansion. Most respondents were of the opinion that oxygen delivery to tissues is of major importance during the management of HRSP. Nigerian consultant anesthetists employ mostly noninvasive blood pressure, CVP, and invasive blood pressure for HM in HRSP. Though a good knowledge of hemodynamic goals was demonstrated, most rated their practice as inadequate.


Rape-related symptoms in adolescents: short- and long-term outcome after cognitive behavior group therapy.
Efficacy studies on treatment in adolescent victims of single rape are lacking, even though sexual victimization is most likely to occur during adolescence and despite the fact that adolescents are at risk to develop subsequent posttraumatic stress disorder. The aim of this prospective observational study was to evaluate the short- and long-term outcomes of a nine-session cognitive behavior group therapy (STEPS), including a parallel six-session parents' group on rape-related symptomatology in female adolescents (13-18 years). STEPS includes psychoeducation, exposure in sensu as well as in vivo, cognitive restructuring, and relapse prevention. Fifty-five female adolescents with mental health problems due to single rape, but without prior sexual trauma, received STEPS while their parents participated in a support group. Subjects were assessed on posttraumatic stress (PTS) and comorbid symptoms using self-report questionnaires prior to and directly after treatment, and at 6 and 12 months follow-up. Repeated measures analysis showed a significant and large decrease in symptoms of PTS, anxiety, depression, anger, dissociation, sexual concerns, and behavior problems directly after treatment, which maintained at 12 months follow-up. Time since trauma did not influence the results. Dropout during STEPS was 1.8%. The results potentially suggest that the positive treatment outcomes at short- and long-term may be caused by STEPS. The encouraging findings need confirmation in future controlled studies on the effectiveness of STEPS because it may be possible that the treatment works especially well for more chronic symptoms, while the less chronic part of the sample showed considerable improvement on its own.


Structure-based energetics of protein interfaces guides foot-and-mouth disease virus vaccine design.
Virus capsids are primed for disassembly, yet capsid integrity is key to generating a protective immune response. Foot-and-mouth disease virus (FMDV) capsids comprise identical pentameric protein subunits held together by tenuous noncovalent interactions and are often unstable. Chemically inactivated or recombinant empty capsids, which could form the basis of future vaccines, are even less stable than live virus. Here we devised a computational method to assess the relative stability of protein-protein interfaces and used it to design improved candidate vaccines for two poorly stable, but globally important, serotypes of FMDV: O and SAT2. We used a restrained molecular dynamics strategy to rank mutations predicted to strengthen the pentamer interfaces and applied the results to produce stabilized capsids. Structural analyses and stability assays confirmed the predictions, and vaccinated animals generated improved neutralizing-antibody responses to stabilized particles compared to parental viruses and wild-type capsids.


Forelimb akinesia in the rat Parkinson model: differential effects of dopamine agonists and nigral transplants as assessed by a new stepping test.
Methods for the assessment of akinesia in the unilateral rat Parkinson model have so far been lacking. The experiments reported here evaluate the usefulness of a new "stepping test" to monitor forelimb akinesia in rats with unilateral 6-hydroxydopamine (6-OHDA) lesions of the mesencephalic dopamine (DA) system, and to assess the ability of DA-receptor agonists and fetal DA neuron transplants to reverse these deficits. The 6-OHDA lesion induced marked and long-lasting impairments in the initiation of stepping movements with the contralateral paw. Systemic injections of low doses (chosen to be subthreshold for induction of rotation) of the mixed D1 and D2 receptor agonist apomorphine, the D1-selective agonist SKF 38393, and to a lesser extent also the D2-selective agonist quinpirole were effective in reversing these deficits. Similar effects was seen after a subrotational dose of L-dopa, whereas amphetamine had no effect. Fetal nigral transplants, implanted as multiple deposits in the ipsilateral caudate-putamen and substantia nigra, restored initiation of stepping to a similar degree as the DA agonists. Nigral grafts placed in substantia nigra alone were also effective, although the improvement was less pronounced. Apomorphine, at a dose effective in the lesion-only animals, had no additive effect in the grafted rats, whereas amphetamine appeared to further improve stepping in the rats with intranigral transplants. Identical experiments were performed on skilled forelimb use in the so-called staircase test. Interestingly, neither the DA agonist drugs nor the nigral transplants had any effects on the lesion induced deficits in this more complex task. The results show that forelimb stepping is a highly useful test to monitor lesion-/and transplant-induced changes in forelimb akinesia, a behavioral parameter that may be analogous to limb akinesia and gait problems seen in patients with Parkinson's disease.


Community matrons: primary care professionals' views and experiences.
This article is a report of a study exploring how the role of the community matron is progressing and any barriers or facilitators to performing the role. Long-term conditions are distressing for patients and costly for healthcare services. Government policy in western countries has increasingly focussed on more integrated and pro-active management of patients with multiple long-term conditions to maintain the ability of the aging population to function independently. The role of community matrons has been introduced in England in an attempt to address these issues. A purposive sample of 31 health and social care professionals participated in five focus group discussions. Reported experiences of the community matron role by community matrons, district nurses, social workers and general practitioners were analysed according to the principles of grounded theory. Data were collected in 2007. The community matron role was seen as effective in meeting the medical and social needs of patients. This was achieved through patient education, developing patients' self-management of their health conditions and monitoring their social needs. Potential barriers to success of the role were associated with lack of role definition prior to its introduction. It is unclear whether the role is financially viable in its current form. Further research is required to examine differing models for implementing the role and judging its effectiveness in bridging the primary-secondary care interface.


[The efficacy of peptide bioregulators of vessels in lower limbs chronic arterial insufficiency treatment in old and elderly people].
Atherosclerosis is a leading cause of obliterating diseases of the arteries in 80-90% of cases. The main pathogenetic mechanism is endothelial dysfunction, leading to hypertension, diabetes, congestive heart failure. The desire of modern society to improve the quality of life increases interest of human health in general and sexual health in particular, because the erectile function is an important factor in the quality of life, which makes us look for more effective methods of diagnosis, treatment and rehabilitation. The objective of the study was to assess the effectiveness of vasoactive tripeptide Vezugen in patients with vasculogenic erectile dysfunction as manifestation of atherosclerosis. Treatment results of erectile dysfunction in 41 patients were studied. A comparative analysis of clinical and instrumental parameters of blood flow in the main penile arteries before and after monotherapy was undertaken. The results show that the blood flow through the main artery of the penis after Vezugen treatment significantly improved both clinical and by objective indicators.


Learning Without Trying: The Clinical Relevance of Statistical Learning.
Statistical learning research seeks to identify the means by which learners, with little perceived effort, acquire the complexities of language. In the past 50 years, numerous studies have uncovered powerful learning mechanisms that allow for learning within minutes of exposure to novel language input. We consider the value of information from statistical learning studies that show potential for making treatment of language disorders faster and more effective. Available studies include experimental research that demonstrates the conditions under which rapid learning is possible, research showing that these findings apply to individuals with disorders, and translational work that has applied learning principles in treatment and educational contexts. In addition, recent research on memory formation has implications for treatment of language deficits. The statistical learning literature offers principles for learning that can improve clinical outcomes for children with language impairment. There is potential for further applications of this basic research that is yet unexplored.


An Overview of Current Methods to Confirm Protein-Protein Interactions.
The research field of protein-protein interactions is interdisciplinary and specialized field that spans all aspects of biology, physics and chemistry. Therefore, in order to discuss the protein-protein interaction in detail and rigorously, it is desirable to integrate knowledge and methods of many related fields including boundary areas such as biochemistry, biophysics and physical chemistry in addition to biology, physics and chemistry. The purpose of this review is to overview current methods to confirm protein-protein interactions. Furthermore, I discuss future prospects of methodology based on current status. It is often necessary to integrate, combine and validate multiple results from various methods to understand protein-protein interactions in detail. It might be desirable for the addition of tags, labeling, and immobilization to solid phases to be unnecessary, and to obtain information on affinity, kinetics, and structure via the analytical method for protein-protein interactions. Therefore, I argue that novel methods based on principles that have already been sufficiently studied in physics or chemistry, but insufficiently applied to the life sciences, should be established to further develop the study of protein-protein interactions.


Streamlining staging of lung and colorectal cancer with whole body MRI; study protocols for two multicentre, non-randomised, single-arm, prospective diagnostic accuracy studies (Streamline C and Streamline L).
Rapid and accurate cancer staging following diagnosis underpins patient management, in particular the identification of distant metastatic disease. Current staging guidelines recommend sequential deployment of various imaging platforms such as computerised tomography (CT) and positron emission tomography (PET) which can be time and resource intensive and onerous for patients. Recent studies demonstrate that whole body magnetic resonance Imaging (WB-MRI) may stage cancer efficiently in a single visit, with potentially greater accuracy than current staging investigations. The Streamline trials aim to evaluate whether WB-MRI increases per patient detection of metastases in non-small cell lung and colorectal cancer compared to standard staging pathways. The Streamline trials are multicentre, non-randomised, single-arm, prospective diagnostic accuracy studies with a novel design to capture patient management decisions during staging pathways. The two trials recruit adult patients with proven or highly suspected new diagnosis of primary colorectal (Streamline C) or non-small cell lung cancer (Streamline L) referred for staging. Patients undergo WB-MRI in addition to standard staging investigations. Strict blinding protocols are enforced for those interpreting the imaging. A first major treatment decision is made by the multi-disciplinary team prior to WB-MRI revelation based on standard staging investigations only, then based on the WB-MRI and any additional tests precipitated by WB-MRI, and finally based on all available test results. The reference standard is derived by a multidisciplinary consensus panel who assess 12 months of follow-up data to adjudicate on the TNM stage at diagnosis. Health psychology assessment of patients' experiences of the cancer staging pathway will be undertaken via interviews and questionnaires. A cost (effectiveness) analysis of WB-MRI compared to standard staging pathways will be performed. We describe a novel approach to radiologist and clinician blinding to ascertain the 'true' diagnostic accuracy of differing imaging pathways and discuss our approach to assessing the impact of WB-MRI on clinical decision making in real-time. The Streamline trials will compare WB-MRI and standard imaging pathways in the same patients, thereby informing the most accurate and efficient approach to staging. Streamline C ISRCTN43958015 (registered 25/7/2012). Streamline L ISRCTN50436483 (registered 31/7/2012).


Polymorphisms of human placental alkaline phosphatase are associated with in vitro fertilization success and recurrent pregnancy loss.
Fertility is a quantitative, complex character governed by a considerable number of genes. Despite clinical and scientific advances, several cases of human infertility remain unexplained. In the present study, using a positional cloning approach in a mouse model of interspecific recombinant lines, a candidate gene, ALPP, encoding the placental alkaline phosphatase, was identified as being potentially involved in recurrent spontaneous abortion. We then analyzed patients for detecting putative associations between ALPP polymorphisms, in vitro fertilization failures, and miscarriages. ALPP was sequenced in 100 controls and 100 patients affected by recurrent spontaneous abortion, from the same ethnic background. The frequency of several alleles and allelic combinations were different between recurrent spontaneous abortion and control women. One polymorphism induced a coding substitution (Ile89Leu) that was associated with a decreased risk of abortion and in vitro fertilization failure. Thereafter, the population was increased by the analysis of 92 additional controls and 612 additional patients for the coding polymorphism Ile89Leu. We finally show, by functional analysis, that the 89Leu placental alkaline phosphatase has an enhanced alkaline phosphatase activity. This study suggests that ALPP genotyping could be a strong predictor of implantation success.


Evolution and expression of the phosphodiesterase 6 genes unveils vertebrate novelty to control photosensitivity.
Phosphodiesterase 6 (PDE6) is a protein complex that hydrolyses cGMP and acts as the effector of the vertebrate phototransduction cascade. The PDE6 holoenzyme consists of catalytic and inhibitory subunits belonging to two unrelated gene families. Rods and cones express distinct genes from both families: PDE6A and PDE6B code for the catalytic and PDE6G the inhibitory subunits in rods while PDE6C codes for the catalytic and PDE6H the inhibitory subunits in cones. We performed phylogenetic and comparative synteny analyses for both gene families in genomes from a broad range of animals. Furthermore, gene expression was investigated in zebrafish. We found that both gene families expanded from one to three members in the two rounds of genome doubling (2R) that occurred at the base of vertebrate evolution. The PDE6 inhibitory subunit gene family appears to be unique to vertebrates and expanded further after the teleost-specific genome doubling (3R). We also describe a new family member that originated in 2R and has been lost in amniotes, which we have named pde6i. Zebrafish has retained two additional copies of the PDE6 inhibitory subunit genes after 3R that are highly conserved, have high amino acid sequence identity, are coexpressed in the same photoreceptor type as their amniote orthologs and, interestingly, show strikingly different daily oscillation in gene expression levels. Together, these data suggest specialisation related to the adaptation to different light intensities during the day-night cycle, most likely maintaining the regulatory function of the PDE inhibitory subunits in the phototransduction cascade.


Preparation, Characterization and Antioxidant Evaluation of Poorly Soluble Polyphenol-Loaded Nanoparticles for Cataract Treatment.
Cataract, one of the leading causes of blindness worldwide, is a condition in which complete or partial opacity develops in the lens of the eyes, thereby impairing vision. This study aimed to examine the potential therapeutic and protective effects of poorly soluble polyphenols like curcumin, resveratrol, and dibenzoylmethane, known to possess significant antioxidant activity. The polyphenols were loaded into novel lipid-cyclodextrin-based nanoparticles and characterized by particle size, polydispersity index, differential scanning calorimetry, thermogravimetric analysis, X-ray diffraction, scanning electron microscopy (SEM), entrapment efficiency, and release studies. Ferric-reducing ability of plasma and 2,2-diphenyl-1-picrylhydrazyl chemical assays were used to evaluate their antioxidant properties based on their free radical quenching ability. Biochemical in vitro assays were used to examine these polyphenols on hydrogen peroxide-induced formation of cataracts in bovine lenses by estimating total glutathione content and superoxide dismutase activity. Nanoparticles were thermostable and amorphous. Particle size of curcumin, resveratrol, and dibenzoylmethane nanoparticles were 331.0 ± 17.9 nm, 329.9 ± 1.9 nm, and 163.8 ± 3.2 nm, respectively. SEM confirmed porous morphology and XRD confirmed physical stability. Entrapment efficiency for curcumin-, resveratrol-, and dibenzoylmethane-loaded nanoparticles was calculated to be 84.4 ± 2.4%, 72.2 ± 1.5%, and 86.4 ± 0.6%, respectively. In vitro release studies showed an initial burst release followed by a continuous release of polyphenols from nanoparticles. Chemical assays confirmed the polyphenols' antioxidant activity. Superoxide dismutase and glutathione levels were found to be significantly increased (p < 0.05) after treatment with polyphenol-loaded nanoparticles than pure polyphenols; thus, an improved antioxidant activity translational into potential anticataract activity of the polyphenols when loaded into nanoparticles was observed as compared to pure polyphenols.


Silkworm silk-based materials and devices generated using bio-nanotechnology.
Silks are natural fibrous protein polymers that are spun by silkworms and spiders. Among silk variants, there has been increasing interest devoted to the silkworm silk of B. mori, due to its availability in large quantities along with its unique material properties. Silk fibroin can be extracted from the cocoons of the B. mori silkworm and combined synergistically with other biomaterials to form biopolymer composites. With the development of recombinant DNA technology, silks can also be rationally designed and synthesized via genetic control. Silk proteins can be processed in aqueous environments into various material formats including films, sponges, electrospun mats and hydrogels. The versatility and sustainability of silk-based materials provides an impressive toolbox for tailoring materials to meet specific applications via eco-friendly approaches. Historically, silkworm silk has been used by the textile industry for thousands of years due to its excellent physical properties, such as lightweight, high mechanical strength, flexibility, and luster. Recently, due to these properties, along with its biocompatibility, biodegradability and non-immunogenicity, silkworm silk has become a candidate for biomedical utility. Further, the FDA has approved silk medical devices for sutures and as a support structure during reconstructive surgery. With increasing needs for implantable and degradable devices, silkworm silk has attracted interest for electronics, photonics for implantable yet degradable medical devices, along with a broader range of utility in different device applications. This Tutorial review summarizes and highlights recent advances in the use of silk-based materials in bio-nanotechnology, with a focus on the fabrication and functionalization methods for in vitro and in vivo applications in the field of tissue engineering, degradable devices and controlled release systems.


Multifeature-based high-resolution palmprint recognition.
Palmprint is a promising biometric feature for use in access control and forensic applications. Previous research on palmprint recognition mainly concentrates on low-resolution (about 100 ppi) palmprints. But for high-security applications (e.g., forensic usage), high-resolution palmprints (500 ppi or higher) are required from which more useful information can be extracted. In this paper, we propose a novel recognition algorithm for high-resolution palmprint. The main contributions of the proposed algorithm include the following: 1) use of multiple features, namely, minutiae, density, orientation, and principal lines, for palmprint recognition to significantly improve the matching performance of the conventional algorithm. 2) Design of a quality-based and adaptive orientation field estimation algorithm which performs better than the existing algorithm in case of regions with a large number of creases. 3) Use of a novel fusion scheme for an identification application which performs better than conventional fusion methods, e.g., weighted sum rule, SVMs, or Neyman-Pearson rule. Besides, we analyze the discriminative power of different feature combinations and find that density is very useful for palmprint recognition. Experimental results on the database containing 14,576 full palmprints show that the proposed algorithm has achieved a good performance. In the case of verification, the recognition system's False Rejection Rate (FRR) is 16 percent, which is 17 percent lower than the best existing algorithm at a False Acceptance Rate (FAR) of 10(-5), while in the identification experiment, the rank-1 live-scan partial palmprint recognition rate is improved from 82.0 to 91.7 percent.


Subcellular drug targeting illuminates local kinase action.
Deciphering how signaling enzymes operate within discrete microenvironments is fundamental to understanding biological processes. A-kinase anchoring proteins (AKAPs) restrict the range of action of protein kinases within intracellular compartments. We exploited the AKAP targeting concept to create genetically encoded platforms that restrain kinase inhibitor drugs at distinct subcellular locations. Local Kinase Inhibition (LoKI) allows us to ascribe organelle-specific functions to broad specificity kinases. Using chemical genetics, super resolution microscopy, and live-cell imaging we discover that centrosomal delivery of Polo-like kinase 1 (Plk1) and Aurora A (AurA) inhibitors attenuates kinase activity, produces spindle defects, and prolongs mitosis. Targeted inhibition of Plk1 in zebrafish embryos illustrates how centrosomal Plk1 underlies mitotic spindle assembly. Inhibition of kinetochore-associated pools of AurA blocks phosphorylation of microtubule-kinetochore components. This versatile precision pharmacology tool enhances investigation of local kinase biology.


Effectiveness of measles vaccination and vitamin A treatment.
The current strategy utilized by WHO/United Nations Children's Fund (UNICEF) to reach the Global Immunization Vision and Strategy 2010 measles reduction goal includes increasing coverage of measles vaccine, vitamin A treatment and supplementation in addition to offering two doses of vaccine to all children. We conducted a systematic review of published randomized controlled trials (RCTs) and quasi-experimental (QE) studies in order to determine effect estimates of measles vaccine and vitamin A treatment for the Lives Saved Tool (LiST). We utilized a standardized abstraction and grading format in order to determine effect estimates for measles mortality employing the standard Child Health Epidemiology Research Group Rules for Evidence Review. We identified three measles vaccine RCTs and two QE studies with data on prevention of measles disease. A meta-analysis of these studies found that vaccination was 85% [95% confidence interval (CI) 83-87] effective in preventing measles disease, which will be used as a proxy for measles mortality in LiST for countries vaccinating before one year of age. The literature also suggests that a conservative 95% effect estimate is reasonable to employ when vaccinating at 1 year or later and 98% for two doses of vaccine based on serology reviews. We included six high-quality RCTs in the meta-analysis of vitamin A treatment of measles which found no significant reduction in measles morality. However, when stratifying by vitamin A treatment dose, at least two doses were found to reduce measles mortality by 62% (95% CI 19-82). Measles vaccine and vitamin A treatment are effective interventions to prevent measles mortality in children.


Contemporary Incidence and Prevalence of Rheumatic Fever and Rheumatic Heart Disease in Australia Using Linked Data: The Case for Policy Change.
Background In 2018, the World Health Organization prioritized control of acute rheumatic fever (ARF) and rheumatic heart disease (RHD), including disease surveillance. We developed strategies for estimating contemporary ARF/RHD incidence and prevalence in Australia (2015-2017) by age group, sex, and region for Indigenous and non-Indigenous Australians based on innovative, direct methods. Methods and Results This population-based study used linked administrative data from 5 Australian jurisdictions. A cohort of ARF (age <45 years) and RHD cases (<55 years) were sourced from jurisdictional ARF/RHD registers, surgical registries, and inpatient data. We developed robust methods for epidemiologic case ascertainment for ARF/RHD. We calculated age-specific and age-standardized incidence and prevalence. Age-standardized rate and prevalence ratios compared disease burden between demographic subgroups. Of 1425 ARF episodes, 72.1% were first-ever, 88.8% in Indigenous people and 78.6% were aged <25 years. The age-standardized ARF first-ever rates were 71.9 and 0.60/100 000 for Indigenous and non-Indigenous populations, respectively (age-standardized rate ratio=124.1; 95% CI, 105.2-146.3). The 2017 Global Burden of Disease RHD prevalent counts for Australia (<55 years) underestimate the burden (1518 versus 6156 Australia-wide extrapolated from our study). The Indigenous age-standardized RHD prevalence (666.3/100 000) was 61.4 times higher (95% CI, 59.3-63.5) than non-Indigenous (10.9/100 000). Female RHD prevalence was double that in males. Regions in northern Australia had the highest rates. Conclusions This study provides the most accurate estimates to date of Australian ARF and RHD rates. The high Indigenous burden necessitates urgent government action. Findings suggest RHD may be underestimated in many high-resource settings. The linked data methods outlined here have potential for global applicability.


CD169⁺ macrophages provide a niche promoting erythropoiesis under homeostasis and stress.
A role for macrophages in erythropoiesis was suggested several decades ago when erythroblastic islands in the bone marrow, composed of a central macrophage surrounded by developing erythroblasts, were described. However, the in vivo role of macrophages in erythropoiesis under homeostatic conditions or in disease remains unclear. We found that specific depletion of CD169(+) macrophages markedly reduced the number of erythroblasts in the bone marrow but did not result in overt anemia under homeostatic conditions, probably because of concomitant alterations in red blood cell clearance. However, CD169(+) macrophage depletion significantly impaired erythropoietic recovery from hemolytic anemia, acute blood loss and myeloablation. Furthermore, macrophage depletion normalized the erythroid compartment in a JAK2(V617F)-driven mouse model of polycythemia vera, suggesting that erythropoiesis in polycythemia vera remains under the control of macrophages in the bone marrow and splenic microenvironments. These results indicate that CD169(+) macrophages promote late erythroid maturation and that modulation of the macrophage compartment may be a new strategy to treat erythropoietic disorders.


Betulinic acid induces apoptosis in skin cancer cells and differentiation in normal human keratinocytes.
Betulinic acid (BA), a pentacyclic triterpene of plant origin, induces cell death in melanoma cells and other malignant cells of neuroectodermal origin. Little is known about additional biological effects in normal target cells. We show, in this study, that BA induces differentiation as well as cell death in normal human keratinocytes (NHK). Cytotoxicity profiles of BA are compared among normal human keratinocytes, HaCaT cells, IGR1 melanoma cells and normal melanocytes. As expected, BA is toxic to all cell types, normal and malignant, but varies in its cytotoxic potency and in the extent of induction of apoptotic vs. necrotic cell death in the four different skin cell types. Apoptosis is proved by annexin V and Apo2.7 binding and by DNA fragmentation. Induction of differentiation-associated antigens in keratinocytes--filaggrin and involucrin--is demonstrated, together with specific morphological changes in treated cell cultures. BA, apart from its cytotoxic activities in cellular systems, is capable of inducing differentiation of NHK into corneocytes without immediately provoking apoptotic cell death.


Platelet dynamics in chronic liver disease with special reference to the role of the spleen.
In all of 12 patients with chronic liver disease, whose platelet dynamics were investigated by the 51Cr-labelling technique in association with surface counting, platelet survival was reduced and in 11 the splenic platelet pool was increased. Surface counting showed high initial spleen liver ratios in eight patients, and in four there was evidence of progressive destruction of platelets in the spleen. In one patient, subsequently shown to have a hepatoma, progressive accumulation of platelets was noted at the tumour site.


Cytotoxic-loaded albumin microspheres: a novel approach to regional chemotherapy.
Adriamycin-loaded albumin microspheres (15-40 micron diameter) incorporating 1 per cent by weight of Adriamycin were prepared by cross linking albumin with glutaraldehyde. After systemic administration in rats, the microspheres were trapped in the pulmonary capillary bed, biodegradation occurring over 48 h. After selective intrarenal arterial injection of 99mTc-labelled albumin microspheres in rabbits, 97 per cent of the injected microspheres were trapped within the kidney; less than 1 per cent were found in the contralateral kidney, lungs and liver. Similar results were obtained after selective intrahepatic arterial injection. The selective injection of cytotoxic-loaded microspheres may provide a reliable method of administering high concentrations of cytotoxic drug to a target organ without producing high serum levels; this technique may have a role in the management of solid tumours.


A phase 1 study of imatinib for corticosteroid-dependent/refractory chronic graft-versus-host disease: response does not correlate with anti-PDGFRA antibodies.
Stimulatory antiplatelet derived growth factor receptor α (PDGFRA) antibodies have been associated with extensive chronic graft-versus-host disease (cGVHD). We performed a phase 1 dose escalation trial of imatinib in corticosteroid-dependent/refractory cGVHD to assess the safety of imatinib and test the hypothesis that abrogation of PDGFRA signaling can ameliorate the manifestations of cGVHD. Fifteen patients were enrolled. Mean follow-up time was 56.6 weeks (range, 18-82.4 weeks). Imatinib 400 mg daily was associated with more frequent moderate to life-threatening adverse events than 200 mg daily. The main adverse events were nausea, edema, confusion, diarrhea, liver function test elevation, fatigue, and myalgia. The overall response rate was 40% (6 of 15). The treatment failure rate was 40% (6 of 15). Twenty percent (3 of 15) of subjects had stable disease. Of 4 subjects with phospho-PDGFRA and phospho-PDGFRB immunohistochemistry studies before and after treatment, inhibition of phosphorylation was observed in 3 but correlated with response in one. Anti-PDGFRA antibodies were observed in 7 of 11 evaluable subjects but correlated with clinical activity in 4. We conclude that cGVHD responds to imatinib through multiple pathways that may include PDGFRA signal transduction. This study is registered at www.clinicaltrials.gov as #NCT00760981.


Prognostic values of electrodiagnostic studies in traumatic radial neuropathy.
It is important to have strong predictors of outcome in traumatic neuropathies so that appropriate management can be instituted early. Our objective in this study was to evaluate the prognostic value of electrodiagnostic studies in traumatic radial neuropathy. In this retrospective study, 33 of 67 subjects with traumatic radial neuropathy met the inclusion criteria. Good outcome was defined as grade 3 or higher strength on the Medical Research Council scale in wrist extensors. Compound muscle action potential (CMAP) responses from extensor indicis proprius (EIP) predicted prognosis: 92% of subjects with a recordable CMAP had a good outcome; and 65% of those with an absent response had a good outcome. Recruitment in brachioradialis was also predictive: 92% of those with full, central, or reduced recruitment had a good outcome; 67% of those with discrete recruitment had a good outcome; and only 33% of those with absent recruitment had a good outcome. Studies performed more than 3 months after injury produced more prognostic certainty than those performed earlier. We conclude that electrodiagnostic studies produce useful prognostic information in traumatic radial neuropathy. It is noteworthy, however, that 65% of subjects with an absent radial CMAP (suggesting complete or nearly complete axon loss) still have a good outcome.