Mean signal and response time influences on multivoxel signals of contextual retrieval in the medial temporal lobe.
The medial temporal lobe supports integrating the "what," "where," and "when" of an experience into a unified memory. However, it remains unclear how representations of these contextual features are neurally encoded and distributed across medial temporal lobe subregions. This study conducted functional magnetic resonance imaging of the medial temporal lobe, while participants retrieved pair, spatial, and temporal source memories. Multivoxel classifiers were trained to distinguish between retrieval conditions before and after correction for mean signal and response times, to more thoroughly characterize the multivoxel signal associated with memory context. Activity in perirhinal and parahippocampal cortex dissociated between memory for associated items and memory for their spatiotemporal context, and hippocampal activity was linked to memory for spatial context. However, perirhinal and hippocampal classifiers were, respectively, driven by effects of mean signal amplitude and task difficulty, whereas the parahippocampal classifier survived correction for these effects. These findings demonstrate dissociable coding mechanisms for episodic memory context across the medial temporal lobe, and further highlight a critical distinction between multivoxel representations driven by spatially distributed activity patterns, and those driven by the regional signal.


Depolarization-induced contractile activity of smooth muscle in calcium-free solution.
In calcium-free solution, strips of cat intestinal muscle developed slow, rhythmic electrical potential changes that triggered contractions. Some strips failed to develop spontaneous electrical activity in calcium-free solution but responded with contractions to depolarization by direct electrical stimulation or by treatment with barium chloride, potassium chloride, or acetylcholine. Similar results were obtained with segments of cat stomach, colon, esophagus, bladder, uterus, and vena cava, as well as with rabbit vena cava. In calcium-free saline, rat small intestinal muscle showed fast electrical activity with accompanying development of a tetanuslike contraction. After 60 min in calcium-free solution, cat small intestinal muscle retained 17.7% of its original concentration of calcium. It is concluded that in some smooth muscles, depolarization-triggered release of intracellular calcium does not require an associated influx of calcium.


Eruption of supernumerary permanent teeth in a sample of urban primary school population in Genoa, Italy.
The aim of this epidemiological study was to describe the incidence and distribution of hyperdontia in the primary school population in Genoa (Italy) and to check its influence on the development of orthodontic problems in children. The collected data should also help to find out what is the best age range among children to direct a program for early diagnosis and prevention of malocclusion and oral diseases related to hyperdontia. The participating children (total number 1577, 814 males and 763 females, between 6 and 10 years of age) chosen in 19 public primary schools in Genoa have been examined by the same specialist through year 2004. Erupted permanent teeth, presence, position and form of supernumerary teeth, malocclusion presence and class, presence of orthodontic devices, age and sex have been noted down for each child. The global percentage of hyperdontia was 0.38%, more frequent in males (0.49%) than in females (0.26%). The most common kind of supernumerary tooth was mesiodens (83%). A significant increase of hyperdontia prevalence (from 0.64% to 1.06%) was noticed in children 9 years old. The incidence of malocclusion among children presenting hyperdontia was 83.3%, while the global incidence of malocclusion was 40%. An orthodontic treatment had been planned and started for 20% of children presenting malocclusion. The study has revealed an incidence of hyperdontia much more frequent in males than in females (2:1). The most common site of eruption of supernumerary teeth is maxillary anterior region. Hyperdontia is strictly related with dental malocclusion. The best age range to direct a program of early diagnosis and prevention of malocclusion and hyperdontia is 9 years old children.


Meta-analysis of trials comparing antidepressants with active placebos.
Unblinding effects may introduce bias into clinical trials. The use of active placebos to mimic side-effects of medication may therefore produce more rigorous evidence on the efficacy of antidepressants. Trials comparing antidepressants with active placebos were located. A standard measure of effect was calculated for each trial and weighted pooled estimates obtained. Heterogeneity was examined and sensitivity analyses performed. A subgroup analysis of in-patient and out-patient trials was conducted. Only two of the nine studies examined produced effect sizes which showed a consistent significant difference in favour of the active drug. Combining all studies produced pooled effect size estimates of between 0.41 (0.27-0.56) and 0.46 (0.31-0.60) with high heterogeneity due to one strongly positive trial. Sensitivity analyses excluding this and one other trial reduced the pooled effect to between 0.21 (0.03-0.38) and 0.27 (0.10-0.45). Meta-analysis is very sensitive to decisions about exclusions. Previous general meta-analyses have found combined effect sizes in the range 0.4-0.8. The more conservative estimates produced here suggest that unblinding effects may inflate the efficacy of antidepressants in trials using inert placebos.


A survey of hepatitis C management by Victorian GPs after PBS-listing of direct-acting antiviral therapy.
To increase access to hepatitis C virus (HCV) treatment, the Pharmaceutical Benefits Scheme (PBS) enabled general practitioners (GPs) to prescribe direct-acting antiviral (DAA) therapy. We conducted a survey to identify GPs' knowledge and management of HCV. A questionnaire consisting of 20 items about HCV knowledge and management was sent to 1000 GPs. One hundred and ninety-one GPs (19.1%) responded; 74% answered correctly that antibody and RNA positivity is diagnostic of HCV. Only 12% could directly request transient elastography. Although 53% of respondents reported interest in prescribing DAAs, 72% continued to refer all patients to specialists. Fifty-five per cent were unsure if people who currently inject drugs were eligible for treatment. Most respondents were interested in prescribing DAAs, but education, access to transient elastography and clear consultation pathways are required to translate this interest into increased treatment availability. PBS eligibility of current injectors needs promotion.


The physiology of blood loss and shock: New insights from a human laboratory model of hemorrhage.
The ability to quickly diagnose hemorrhagic shock is critical for favorable patient outcomes. Therefore, it is important to understand the time course and involvement of the various physiological mechanisms that are active during volume loss and that have the ability to stave off hemodynamic collapse. This review provides new insights about the physiology that underlies blood loss and shock in humans through the development of a simulated model of hemorrhage using lower body negative pressure. In this review, we present controlled experimental results through utilization of the lower body negative pressure human hemorrhage model that provide novel insights on the integration of physiological mechanisms critical to the compensation for volume loss. We provide data obtained from more than 250 human experiments to classify human subjects into two distinct groups: those who have a high tolerance and can compensate well for reduced central blood volume (e.g. hemorrhage) and those with low tolerance with poor capacity to compensate.We include the conceptual introduction of arterial pressure and cerebral blood flow oscillations, reflex-mediated autonomic and neuroendocrine responses, and respiration that function to protect adequate tissue oxygenation through adjustments in cardiac output and peripheral vascular resistance. Finally, unique time course data are presented that describe mechanistic events associated with the rapid onset of hemodynamic failure (i.e. decompensatory shock). Impact Statement Hemorrhage is the leading cause of death in both civilian and military trauma. The work submitted in this review is important because it advances the understanding of mechanisms that contribute to the total integrated physiological compensations for inadequate tissue oxygenation (i.e. shock) that arise from hemorrhage. Unlike an animal model, we introduce the utilization of lower body negative pressure as a noninvasive model that allows for the study of progressive reductions in central blood volume similar to those reported during actual hemorrhage in conscious humans to the onset of hemodynamic decompensation (i.e. early phase of decompensatory shock), and is repeatable in the same subject. Understanding the fundamental underlying physiology of human hemorrhage helps to test paradigms of critical care medicine, and identify and develop novel clinical practices and technologies for advanced diagnostics and therapeutics in patients with life-threatening blood loss.


Prospective use of intramuscular triamcinolone acetonide in pseudogout.
To prospectively assess the efficacy of intramuscular (i.m.) triamcinolone acetonide in the treatment of pseudogout. Fourteen patients with crystal proven pseudogout presenting with an acute attack within 5 days of onset were treated with intramuscular triamcinolone acetonide 60 mg and followed for 30 days. Patients with inadequate response were eligible for a 2nd triamcinolone acetonide injection on Day 1-2. Twelve patients had contraindication to nonsteroidal antiinflammatory agents (NSAID). Acute arthritis was monoarticular in 10 patients, and involved 2 or more joints in 4 patients. All patients had good clinical response to triamcinolone acetonide based on restoration of near baseline joint range of motion and joint circumference, and at least 50% improvement in patient and physician global assessment. Major clinical improvement occurred by Day 1-2 (2 patients), Day 3-4 (11 patients), and Day 10-14 (one patient). Six patients required a 2nd triamcinolone acetonide injection on Day 1-2. Toxicities were not observed. I.m. triamcinolone acetonide appears to be safe, well tolerated, and effective in the treatment of pseudogout. It may be a reasonable alternative therapy when NSAID are contraindicated, and for polyarticular attacks where intraarticular corticosteroids are impractical.


Temporal endogenous gene expression profiles in response to polymer-mediated transfection and profile comparison to lipid-mediated transfection.
Design of efficient nonviral gene delivery systems is limited by the rudimentary understanding of specific molecules that facilitate transfection. Polyplexes using 25-kDa polyethylenimine (PEI) and plasmid-encoding green fluorescent protein (GFP) were delivered to HEK 293T cells. After treating cells with polyplexes, microarrays were used to identify endogenous genes differentially expressed between treated and untreated cells (2 h of exposure) or between flow-separated transfected cells (GFP+) and treated, untransfected cells (GFP-) at 8, 16 and 24 h after lipoplex treatment. Cell priming studies were conducted using pharmacologic agents to alter endogenous levels of the identified differentially expressed genes to determine effect on transfection levels. Differentially expressed genes in polyplex-mediated transfection were compared with those differentially expressed in lipoplex transfection to identify DNA carrier-dependent molecular factors. Differentially expressed genes were RGS1, ARHGAP24, PDZD2, SNX24, GSN and IGF2BP1 after 2 h; RAP1A and ACTA1 after 8 h; RAP1A, WDR78 and ACTA1 after 16 h; and RAP1A, SCG5, ATF3, IREB2 and ACTA1 after 24 h. Pharmacologic studies altering endogenous levels for ARHGAP24, GSN, IGF2BP1, PDZD2 and RGS1 were able to increase or decrease transgene production. Comparing differentially expressed genes for polyplexes and lipoplexes, no common genes were identified at the 2-h time point, whereas, after the 8-h time point, RAP1A, ATF3 and HSPA6 were similarly expressed. SCG5 and PGAP1 were only upregulated in polyplex-transfected cells. The identified genes and pharmacologic agents provide targets for improving transfection systems, although polyplex or lipoplex dependencies must be considered.


Magnetized suspended carbon nanotubes based nanofluid flow with bio-convection and entropy generation past a vertical cone.
The captivating attributes of carbon nanotubes (CNT) comprising chemical and mechanical steadiness, outstanding electrical and thermal conductivities, featherweight, and physiochemical consistency make them coveted materials in the manufacturing of electrochemical devices. Keeping in view such exciting features of carbon nanotubes, our objective in the present study is to examine the flow of aqueous based nanofluid comprising single and multi-wall carbon nanotubes (CNTs) past a vertical cone encapsulated in a permeable medium with convective heat and solutal stratification. The impacts of heat generation/absorption, gyrotactic-microorganism, thermal radiation, and Joule heating with chemical reaction are added features towards the novelty of the erected model. The coupled differential equations are attained from the partial differential equations by exercising the local similarity transformation technique. The set of conservation equations supported by the associated boundary conditions are worked out numerically by employing bvp4c MATLAB function. The sway of numerous appearing parameters in the analysis on the allied distributions is scrutinized and the fallouts are portrayed graphically. The physical quantities of interest including Skin friction coefficient, the rate of heat and mass transfers are assessed versus essential parameters and their outcomes are demonstrated in tabulated form. It is witnessed that the velocity of the fluid decreases for boosting values of the magnetic and suction parameters in case of both nanotubes. Moreover, the density of motile microorganism is decreased versus larger estimates of bio-convection constant. A notable highlight of the presented model is the endorsement of the results by matching them to an already published material in the literature. A venerable harmony in this regard is achieved.


Object-related regularities are processed automatically: evidence from the visual mismatch negativity.
One of the most challenging tasks of our visual systems is to structure and integrate the enormous amount of incoming information into distinct coherent objects. It is an ongoing debate whether or not the formation of visual objects requires attention. Implicit behavioral measures suggest that object formation can occur for task-irrelevant and unattended visual stimuli. The present study investigated pre-attentive visual object formation by combining implicit behavioral measures and an electrophysiological indicator of pre-attentive visual irregularity detection, the visual mismatch negativity (vMMN) of the event-related potential. Our displays consisted of two symmetrically arranged, task-irrelevant ellipses, the objects. In addition, there were two discs of either high or low luminance presented on the objects, which served as targets. Participants had to indicate whether the targets were of the same or different luminance. In separate conditions, the targets either usually were enclosed in the same object or in two different objects (standards). Occasionally, the regular target-to-object assignment was changed (deviants). That is, standards and deviants were exclusively defined on the basis of the task-irrelevant target-to-object assignment but not on the basis of some feature regularity. Although participants did not notice the regularity nor the occurrence of the deviation in the sequences, task-irrelevant deviations resulted in increased reaction times. Moreover, compared with physically identical standard displays deviating target-to-object assignments elicited a negative potential in the 246-280 ms time window over posterio-temporal electrode positions which was identified as vMMN. With variable resolution electromagnetic tomography (VARETA) object-related vMMN was localized to the inferior temporal gyrus. Our results support the notion that the visual system automatically structures even task-irrelevant aspects of the incoming information into objects.


Role and modulation of the secondary structure of antimicrobial peptides to improve selectivity.
Since the development of bacterial resistance, the decreasing effectiveness of antibiotics is becoming one of the most critical problems worldwide. Novel antibacterial agents are urgently needed to prevent humanity from falling back into the "post-antibiotic era". As an important part of the innate immune system, antimicrobial peptides (AMPs) are one of the most promising antibacterial agents showing broad-spectrum activity against bacteria and low propensity for drug resistance. However, the shortcomings of AMPs, such as high toxicity and easy digestion by proteases, limit their clinical application. This review mainly focuses on the effect of the secondary structure on the antimicrobial activity and cytotoxicity of AMPs and the strategies of designing conformationally transitionable AMPs with improved selectivity towards bacteria.


Tacrolimus and 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors: An interaction study in CYP3A5 non-expressors, renal transplant recipients.
Atherosclerosis is a significant factor affecting long-term outcome in renal transplant recipients. Studies have been conducted to determine the pharmacogenomic pathways involved in statin efficacy, efficiency, and adverse effect likelihood. However, little is known about the influence of statins on tacrolimus kinetics. The aim of this study was to investigate possible pharmacological interactions between tacrolimus and statins in CYP3A5 non-expressors, renal transplant recipients. Twenty-four patients, treated with tacrolimus (n=24), methylprednisolone (n=24), and mycophenolate mofetil (n=19)/azathioprine (n=1)/everolimus (n=4), participated in the study. After an observation time of 112±36 days, statins, namely, atorvastatin (n=12), simvastatin (n=8), pravastatin (n=2), or fluvastatin (n=2), were administered for additional 101±34 days. DNA was extracted from whole blood sample and polymerase chain reaction followed by restriction fragment length polymorphism analysis was used for CYP3A5 genotyping. Student's t-test and Mann-Whitney test were used to test the significance of difference in variables that passed or did not pass Kolmogorov's normality test, respectively. No statistically significant difference was observed in tacrolimus daily dose, concentration, concentration/dose ratio, and volume of distribution before and during the administration of statins. Statistically significant decrease in serum cholesterol was observed after initiation of statins. Renal and hepatic function remained unchanged and no skeletal muscle abnormalities were reported. The results of this study show that tacrolimus and statins do not interact in terms of efficacy, efficiency, and adverse effect likelihood. No significant clinical interaction or effect was observed, even with the use of atorvastatin or simvastatin, which are metabolized by CYP3A4 such as tacrolimus.


Localization and regulation of reproductive steroid receptors in the raphe serotonin system of male macaques.
We previously showed that tryptophan hydroxylase 2 (TPH2) and serotonin reuptake transporter (SERT) mRNAs are increased by the androgens, testosterone (T) and dihydrotestosterone (DHT) in serotonin neurons of male macaques. In addition, we observed that serotonin in axons of a terminal region were markedly decreased by aromatase inhibition and lack of estradiol (E) from metabolism of T. These observations implicated androgen receptors (AR) and estrogen receptors (ER) in the transduction of steroid hormone actions in serotonin neurons. Due to the longer treatment period employed, the expression of the cognate nuclear receptors was sought. We used single and double immunohistochemistry to quantitate and phenotypically localize AR, ERα and ERβ in the dorsal raphe of male macaques. Male Japanese macaques (Macaca fuscata) were castrated for 5-7 months and then treated for 3 months with [1] placebo, [2] T, [3] DHT (non-aromatizable androgen) plus ATD (steroidal aromatase inhibitor), or [4] Flutamide (FLUT; androgen antagonist) plus ATD (n = 5/group). After single labeling of each receptor, quantitative image analysis was applied and receptor positive neurons were counted. Double-label of raphe neurons for each receptor plus TPH2 determined whether the receptors were localized in serotonin neurons. There were significantly more AR-positive neurons in T- and DHT+ATD-treated groups (p = 0.0014) compared to placebo or FLUT+ATD-treated groups. There was no difference in the number of positive-neurons stained for ERα or ERβ⋅ Double-immunohistochemistry revealed that serotonin neurons did not contain AR. Rather, AR-positive nuclei were found in neighboring cells that are likely neurons. However, approximately 40% of dorsal raphe serotonin neurons contained ERα or ERβ⋅ In conclusion, the stimulatory effect of androgens on TPH2 and SERT mRNA expression is mediated indirectly by neighboring neurons contain AR. The stimulatory effect of E, derived from T metabolism, on serotonin transport is partially mediated directly via nuclear ERs.


Cytokine Secretion, Viability, and Real-Time Proliferation of Apical-Papilla Stem Cells Upon Exposure to Oral Bacteria.
The use of stem cells from the apical papilla (SCAPs) has been proposed as a means of promoting root maturation in permanent immature teeth, and plays a significant role in regenerative dental procedures. However, the role of SCAPs may be compromised by microenvironmental factors, such as hypoxic conditions and the presence of bacteria from infected dental root canals. We aim to investigate oral bacterial modulation of SCAP in terms of binding capacity using flow cytometry and imaging, real-time cell proliferation monitoring, and cytokine secretion (IL-6, IL-8, and TGF-β isoforms) under anaerobic conditions. SCAPs were exposed to key species in dental root canal infection, namely Actinomyces gerensceriae, Slackia exigua, Fusobacterium nucleatum, and Enterococcus faecalis, as well as two probiotic strains, Lactobacillus gasseri strain B6 and Lactobacillus reuteri (DSM 17938). We found that A. gerensceriae, S. exigua, F. nucleatum, and E. faecalis, but not the Lactobacillus probiotic strains bind to SCAPs on anaerobic conditions. Enterococcus faecalis and F. nucleatum exhibited the strongest binding capacity, resulting in significantly reduced SCAP proliferation. Notably, F. nucleatum, but not E. faecalis, induce production of the proinflammatory chemokine IL-8 and IL-10 from SCAPs. Production of TGF-β1 and TGF-β2 by SCAPs was dependent on species, cell line, and time, but secretion of TGF-β3 did not vary significantly over time. In conclusion, SCAP response is compromised when exposed to bacterial stimuli from infected dental root canals in anaerobic conditions. Thus, stem cell-mediated endodontic regenerative studies need to include microenvironmental conditions, such as the presence of microorganisms to promote further advantage in the field.


Tailoring Eco-Friendly Colloidal Quantum Dots for Photoelectrochemical Hydrogen Generation.
A photoelectrochemical (PEC) cell is able to realize effective solar-to-hydrogen energy conversion from water by using the semiconductor photoelectrode. Semiconducting colloidal quantum dots (QDs) with captivating features of size-tunable optoelectronic properties and broad light absorption are regarded as promising photosensitizers in solar-driven PEC systems. Up to now, different types of QDs have been developed to achieve high-efficiency PEC H2 generation, while the majority of state-of-the-art QDs-PEC systems are still fabricated from QDs consisting of heavy metals (e.g., Cd and Pb), which are extremely harmful to the human health and natural environment. In this context, substantial efforts have been made to mitigate the usage of highly toxic heavy metals and concurrently promote the development of alternative environment-friendly QDs with comparable features. This review presents recent advances of solar-driven PEC devices based on several typical environment-friendly QDs (e.g., carbon QDs, I-III-VI QDs and III-V QDs). A variety of techniques (e.g., shell thickness tuning, alloying/doping, and ligands exchange, etc.) to engineer these QD's optoelectronic properties and achieve high-efficiency PEC H2 production are thoroughly discussed. Furthermore, the critical challenges and future perspectives of advanced eco-friendly QDs-PEC systems in terms of QDs' synthesis, photo-induced charge kinetics, and operation stability/efficiency are briefly proposed.


Silver toxicity with the use of silver-impregnated dressing and wound vacuum-assisted closure in an immunocompromised patient.
Silver-containing topical agents are used to help prevent infectious complications in wound therapy. Toxicity from topical silver agent exposure was initially reported in 1975 and was clinically characterized by granulocytopenia. Currently, the data regarding potential toxicity associated with silver-impregnated devices are limited. A 23-year-old patient receiving chemotherapy for acute lymphoblastic leukemia presented with necrotizing fasciitis of the abdominal wall and scrotum from a Crohn disease-related psoas-enteric fistula. Surgical debridement of the soft-tissue and abdominal musculature was performed to the peritoneum. Silver-containing foam sponges and wound vacuum-assisted closure were applied directly to the peritoneum 2 weeks after initial debridement. Subsequently, the patient developed leukopenia, and workup revealed the serum silver level was 4 times normal level. Silver-impregnated sponges were discontinued and silver-free sponges and wound vacuum-assisted closure therapy resumed, followed by leukopenia resolution. Silver toxicity associated with routine application of silver-impregnated sponges has not been previously reported.


An ancestral role in peroxisome assembly is retained by the divisional peroxin Pex11 in the yeast Yarrowia lipolytica.
The peroxin Pex11 has a recognized role in peroxisome division. Pex11p remodels and elongates peroxisomal membranes prior to the recruitment of dynamin-related GTPases that act in membrane scission to divide peroxisomes. We performed a comprehensive comparative genomics survey to understand the significance of the evolution of the Pex11 protein family in yeast and other eukaryotes. Pex11p is highly conserved and ancestral, and has undergone numerous lineage-specific duplications, whereas other Pex11 protein family members are fungal-specific innovations. Functional characterization of the in-silico-predicted Pex11 protein family members of the yeast Yarrowia lipolytica, i.e. Pex11p, Pex11Cp and Pex11/25p, demonstrated that Pex11Cp and Pex11/25p have a role in the regulation of peroxisome size and number characteristic of Pex11 protein family members. Unexpectedly, deletion of PEX11 in Y. lipolytica produces cells that lack morphologically identifiable peroxisomes, mislocalize peroxisomal matrix proteins and preferentially degrade peroxisomal membrane proteins, i.e. they exhibit the classical pex mutant phenotype, which has not been observed previously in cells deleted for the PEX11 gene. Our results are consistent with an unprecedented role for Pex11p in de novo peroxisome assembly.


The Caenorhabiditis elegans model as a reliable tool in neurotoxicology.
Caenorhabiditis elegans (C. elegans) offers an attractive experimental platform as it has a short life cycle, is inexpensive to maintain and most importantly has high degree of evolutionary conservation with higher eukaryotes. Understanding the contribution of inherent genes that regulate neurotoxicity and antioxidant stress responses in the worm provides critical insight into mechanisms of mammalian neurotoxicity. The C. elegans model readily enables multi-gene approach, allowing for combinatorial genetic variation to be studied within the context of the influence of multigenic polymorphisms in environmental risk and vulnerability. This review provides a synopsis of recent studies on metal and pesticides toxicity in C. elegans, highlighting the utility of the model system in understanding molecular mechanisms that underlie developmental, reproductive and neuronal damage. The continuation of these investigations combining basic toxicological experimentation with novel genetic and high throughput methods will continue to make C. elegans an invaluable tool for future research, providing insight into molecular and cellular mechanisms of toxicity.


Behavior of epidermolysis bullosa fibroblasts in a hydrated collagen lattice.
To determine if an altered ability to contract a hydrated collagen lattice is characteristic of fibroblasts from patients with recessive dystrophic epidermolysis bullosa (RDEB), we examined contraction by fibroblasts from normal subjects and patients with RDEB, dominant dystrophic epidermolysis bullosa (DDEB), and dominant epidermolysis bullosa simplex (DEBS). An extremely broad range of contractility (normal, poor, and hypercontraction) was observed in all types of epidermolysis bullosa (EB). When contraction in control fibroblasts was defined as the mean +/- 2 SD, (all control values were within this range) and the data were analyzed by the chi-square test, only 32% of EB cells fell within this range, with 47% poorly contractile and 21% hypercontractile. These data, derived from 34 patients, indicate that no single genetic defect resulting in altered contractility in the 3 distinct types of EB is likely. Neither cell viability, collagenase expression, nor PGE2 synthesis as correlated with gel contraction in any group. Indomethacin had no effect on contraction in RDEB. It is possible that the genetic defects in EB cause blister formation in vivo and may lead in some way to an abnormal interaction of fibroblasts with the extracellular matrix resulting in an altered collagen lattice contraction in vitro.


Individual determinants of research utilization: a systematic review.
In order to design interventions that increase research use in nursing, it is necessary to have an understanding of what influences research use. To report findings on a systematic review of studies that examine individual characteristics of nurses and how they influence the utilization of research. A survey of published articles in English that examine the influence of individual factors on the research utilization behaviour of nurses, without restriction of the study design, from selected computerized databases and hand searches. Articles had to measure one or more individual determinants of research utilization, measure the dependent variable (research utilization), and evaluate the relationship between the dependent and independent variables. The studies also had to indicate the direction of the relationship between the independent and dependent variables, report a P-value and the statistic used, and indicate the magnitude of the relationship. Six categories of potential individual determinants were identified: beliefs and attitudes, involvement in research activities, information seeking, professional characteristics, education and other socio-economic factors. Research design, sampling, measurement, and statistical analysis were examined to evaluate methodological quality. Methodological problems surfaced in all of the studies and, apart from attitude to research, there was little to suggest that any potential individual determinant influences research use. Important conceptual and measurement issues with regard to research utilization could be better addressed if research in the area were undertaken longitudinally by multi-disciplinary teams of researchers.


Novel small molecule XPO1/CRM1 inhibitors induce nuclear accumulation of TP53, phosphorylated MAPK and apoptosis in human melanoma cells.
XPO1/CRM1 is a key nuclear exporter protein that mediates translocation of numerous cellular regulatory proteins. We investigated whether XPO1 is a potential therapeutic target in melanoma using novel selective inhibitors of nuclear export (SINE). In vitro effects of SINE on cell growth and apoptosis were measured by MTS assay and flow cytometry [Annexin V/propidium iodide (PI)], respectively in human metastatic melanoma cell lines. Immunoblot analysis was used to measure nuclear localization of key cellular proteins. The in vivo activity of oral SINE was evaluated in NOD/SCID mice bearing A375 or CHL-1 human melanoma xenografts. SINE compounds induced cytostatic and pro-apoptotic effects in both BRAF wild type and mutant (V600E) cell lines at nanomolar concentrations. The cytostatic and pro-apoptotic effects of XPO1 inhibition were associated with nuclear accumulation of TP53, and CDKN1A induction in the A375 cell line with wild type TP53, while pMAPK accumulated in the nucleus regardless of TP53 status. The orally bioavailable KPT-276 and KPT-330 compounds significantly inhibited growth of A375 (p<0.0001) and CHL-1 (p = 0.0087) human melanoma cell lines in vivo at well tolerated doses. Inhibition of XPO1 using SINE represents a potential therapeutic approach for melanoma across cells with diverse molecular phenotypes by promoting growth inhibition and apoptosis.


Exhaled Carbon Monoxide Levels in Forty Resistant to Cessation Male Smokers after Six Months of Full Switch to Electronic Cigarettes (e-Cigs) or to A Tobacco Heating Systems (THS).
Cigarette smoke releases several toxic chemicals and carcinogens including carbon monoxide (CO). This study examined the levels of exhaled CO in smokers switching to electronic cigarettes (e-Cigs) or a tobacco heating system (THS) and their level of compliance six months after switching. On the basis of their own preferences, 40 male smokers unwilling or unable to stop smoking were switched to e-Cigs or THSs for six months (20 subjects in each group). Nicotine addiction and levels of carbon monoxide in the exhaled breath (eCO) were measured at baseline (the latter also at six months). The Shapiro Wilk test, graphical methods, Student T test or Mann-Whitney test were used to assess the normal distribution of variables and differences between the two groups after six months. The two groups showed no difference at baseline, but a significant higher addiction score in smokers choosing THS. E-Cig and THS showed significant reduced levels of eCO (both %COHb and COppm) after six months, which were within the range of non-smoker status. Reduced levels of %COHb did not significantly differ between the two groups, whilst the THS group had a significantly lower reduction in levels of COppm vs the e-Cig group (p < 0.05). Both e-Cigs and THSs are capable of significantly reducing eCO at least in the medium term, hence constituting a viable tobacco harm reduction approach in smokers who are unwilling or unable to stop smoking.


Treatment of murine invasive candidiasis with amphotericin B and cilofungin: evidence for enhanced activity with combination therapy.
The in vivo interactions of cilofungin, an echinocandin antifungal agent, and amphotericin B, a polyene derivative, in a murine model of disseminated candidiasis have been investigated. While single therapy with either drug alone prolonged survival of infected mice, kidney colony counts were not appreciably reduced. In contrast, combination therapy, especially at higher doses of both drugs, resulted in significant prolongation of survival and suppression of growth of yeast cells in the kidneys. Combination therapy of experimental candidiasis with cilofungin and amphotericin B did not result in antagonism; rather, additive or synergistic effects were seen. Future preclinical work with other echinocandin and polyene derivatives should include studies evaluating the in vivo interactions of both classes of compounds.


The pediatrician workforce: current status and future prospects.
The effective and efficient delivery of children's health care depends on the pediatrician workforce. The number, composition, and distribution of pediatricians necessary to deliver this care have been the subject of long-standing policy and professional debate. This technical report reviews current characteristics and recent trends in the pediatric workforce and couples the workforce to a conceptual model of improvement in children's health and well-being. Important recent changes in the workforce include (1) the growth in the number of pediatricians in relation to the child population, (2) increased numbers of female pediatricians and their attainment of majority gender status in the specialty, (3) the persistence of a large number of international medical graduates entering training programs, (4) a lack of ethnic and racial diversity in pediatricians compared with children, and (5) the persistence of marked regional variation in pediatrician supply. Supply models projecting the pediatric workforce are reviewed and generally indicate that the number of pediatricians per child will increase by 50% over the next 20 years. The differing methods of assessing workforce requirements are presented and critiqued. The report finds that the pediatric workforce is undergoing fundamental changes that will have important effects on the professional lives of pediatricians and children's health care delivery.


The Role of Clearinghouses in Promoting Transparent Research: A Methodological Study of Transparency Practices for Preventive Interventions.
Transparency of research methods is vital to science, though incentives are variable, with only some journals and funders adopting transparency policies. Clearinghouses are also important stakeholders; however, to date none have implemented formal procedures that facilitate transparent research. Using data from the longest standing clearinghouse, we examine transparency practices for preventive interventions to explore the role of online clearinghouses in incentivizing researchers to make their research more transparent. We conducted a descriptive analysis of 88 evaluation reports reviewed in 2018-2019 by Blueprints for Healthy Youth Development, when the clearinghouse began checking for trial registrations, and expanded on these efforts by applying broader transparency standards to interventions eligible for an endorsement on the Blueprints website during the study period. Reports were recent, with 84% published between 2010 and 2019. We found that few reports had data, code, or research materials that were publicly available. Meanwhile, 40% had protocols that were registered, but only 8% were registered prospectively, while one-quarter were registered before conducting analyses. About one-third included details in a registered protocol describing the treatment contrast and planned inclusions, and less than 5% had a registered statistical analysis plan (e.g., planned analytical methods, pre-specified covariates). Confirmatory research was distinguished from exploratory work in roughly 40% of reports. Reports published more recently (after 2015) had higher rates of transparency. Preventive intervention research needs to be more transparent. Since clearinghouses rely on robust findings to make well-informed decisions and researchers are incentivized to meet clearinghouse standards, clearinghouses should consider policies that encourage transparency to improve the credibility of evidence-based interventions.


Prospective histopathologic evaluation of lifestyle modification in nonalcoholic fatty liver disease: a randomized trial.
Nonalcoholic fatty liver disease (NAFLD) is now recognized as part of the metabolic syndrome, and is specifically related to obesity and insulin resistance. Lifestyle modification is advocated for the treatment of NAFLD, but few studies have evaluated its impact on liver histology. The purpose of this study was to investigate which, if any, specific diet and exercise recommendations are associated with histopathologic changes. A total of 56 participants were randomly assigned to 1 of 4 lifestyle modification subgroups for 6 months: standard care, low-fat diet and moderate exercise, moderate-fat/low-processed-carbohydrate diet and moderate exercise, or moderate exercise only. All subjects had biopsy-proven NAFLD, to include nonalcoholic steatohepatitis (NASH), and received a repeat 6-month biopsy to detect histopathologic changes. Other measures included blood assay of liver enzymes (aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase), fasting glucose, serum insulin, lipid panel, body weight, dietary intake, fat mass, and fitness level. Among the 41 participants who completed the study (88% with NASH), a significant change was found in pre- to post-NAFLD activity score in the group as a whole (p < 0.001) with no difference detected between subgroups (p = 0.31). Our results confirm that lifestyle modification is effective in improving NAFLD and NASH. Regardless of intervention group, lifestyle modification improved liver histology, as verified by repeat biopsy, after a 6-month intervention. This study reinforces the importance of lifestyle modification as the primary treatment strategy for patients with NAFLD.


Irreversible Electroporation for De-epithelialization of Murine Small Intestine.
Nonthermal irreversible electroporation (NTIRE) has been shown to ablate the small intestinal epithelium while maintaining submucosal and muscularis propriae integrity. NTIRE is used here in a first-in-mouse study to eliminate the native intestinal stem cell population to understand optimal parameters and timeline of mucosal regeneration. Adult C57 background mice underwent laparotomy and electroporation of 1.5 cm of jejunum using a BTX 830 ECM electroporator and electrode calipers. Parameters were varied by voltage, pulse number, interval, and duration to determine optimal de-epithelialization. Electroporated segments were extracted 1 to 3 d after intervention with same-animal control segment. Cross sections were preserved, and measurements were taken to compare effects of parameters on villi height, crypt depth, crypt obliteration, and serosal thickness. Morbidity was limited at a standard set of electroporation parameters (14%), and increased with higher voltage, longer interval, and shorter or longer pulses. Serosa/muscularis thickness was unaffected by varying interventions. Crypt depth and obliterated crypts were most affected by modulating pulse number, intervals, and duration. Villi height was most significantly shortened by altering pulse duration, with limited recovery by day 3, otherwise mucosal regeneration was observed in most cases by this point. NTIRE is an effective method of denuding small intestinal epithelium in mice and temporarily ablating crypts while sparing the support scaffold for native regeneration. This first-in-mouse study of electroporation suggests it is a practical tool that can be utilized in a small mammalian system.


The incidence of nerve root injury by high-speed drill can be reduced by chilled saline irrigation in a rabbit model.
We aimed to evaluate the temperature around the nerve root during drilling of the lamina and to determine whether irrigation during drilling can reduce the chance of nerve root injury. Lumbar nerve roots were exposed to frictional heat by high-speed drilling of the lamina in a live rabbit model, with saline (room temperature (RT) or chilled saline) or without saline (control) irrigation. We measured temperatures surrounding the nerve root and made histological evaluations. In the control group, the mean temperature around the nerve root was 52.0°C (38.0°C to 75.5°C) after 60 seconds of drilling, and nerve root injuries were found in one out of 13 (7.7%) immediately, three out of 14 (21.4%) at three days, and 11 out of 25 (44.0%) at seven days post-operatively. While the RT group showed a significantly lower temperature around the nerve root compared with the control group (mean 46.5°C; 34.5°C to 66.9°C, p < 0.001), RT saline failed to significantly reduce the incidence of nerve root injury (ten out of 26; 38.5%; odds ratio (OR) 0.96; 95% confidence interval (CI) 0.516 to 1.785; p = 0.563). However, chilled saline irrigation resulted in a significantly lower temperature than the control group (mean 39.0°C; 35.3°C to 52.3°C; p < 0.001) and a lower rate of nerve root injury (two out of 21; 9.5%, OR 0.13; 95% CI 0.02 to 0.703, p = 0.010). Frictional heat caused by a high-speed drill can cause histological nerve root injury. Chilled saline irrigation had a more prominent effect than RT in reducing the incidence of the thermal injury during extended drilling. Cite this article: Bone Joint J 2017;99-B:554-60.


Interpersonal influences on decision-making capacity: a content analysis of court judgments.
For many purposes in England and Wales, the Court of Protection determines whether a person has or lacks capacity to make a decision, by applying the test within the Mental Capacity Act 2005. This test is regularly described as a cognitive test with cognitive processes discussed as internal characteristics. However, it is unclear how the courts have framed interpersonal influence as negatively impacting upon a person's decision-making processes in a capacity assessment context. We reviewed published court judgments in England and Wales in which interpersonal problems were discussed as relevant to capacity. Through content analysis, we developed a typology that highlights five ways the courts considered influence to be problematic to capacity across these cases. Interpersonal influence problems were constructed as (i) P's inability to preserve their free will or independence, (ii) restricting P's perspective, (iii) valuing or dependence on a relationship, (iv) acting on a general suggestibility to influence, or (v) P denying facts about the relationship. These supposed mechanisms of interpersonal influence problems are poorly understood and clearly merit further consideration. Our typology and case discussion are a start towards more detailed practice guidelines, and raise questions as to whether mental capacity and influence should remain legally distinct.


Acanthamoeba keratitis. A review of the literature.
Acanthamoeba is a free-living ubiquitous ameba that is responsible for a small but increasing number of cases of keratitis. The infection is associated with minimal corneal trauma and soft contact lens wear. It typically presents as a unilateral central or paracentral corneal infiltrate, often with a ring-shaped peripheral infiltrate. The lesion is often confused with fungal, bacterial, or herpetic keratitis. Successful therapy hinges on early recognition and aggressive therapy with appropriate topical antiamebic medication often in conjunction with penetrating keratoplasty. Thirty-five cases from the world literature are reviewed.


[The effects of TSH, cholera toxin and Graves' IgG on cAMP production in cultured human thyroid adenoma cells in monolayer].
Monolayer cultures of human thyroid cells derived from thyroid adenoma were utilized for the assay of thyroid stimulating substances such as thyrotropin (TSH), cholera toxin and thyroid stimulating immunoglobulin (TSI) in patients with Graves' disease. Adenoma cells were treated with 0.1% collagenase or 2000 unit/ml dispase to thyrocytes. The cells were cultured in MEM containing 10% fetal calf serum under an atmosphere of 5% CO2 in air. Within 24 hours, the cells attached themselves to the plastic surface and formed a monolayer. Cyclic AMP responses to TSH, cholera toxin or Graves' IgG were tested in a medium (PBS) containing 0.5 mM IBMX. The cyclic AMP responses to TSH were generally maximal on the 3rd day of culture and declined thereafter. The response was dose-dependent, and 10 microU/ml of TSH produced a significant increase of cellular cyclic AMP. The response by 1 microU/ml of TSH was 28 approximately 57 fold above the basal. The response was also a function of the incubation period. The maximal response was attained after 1 h incubation. When the cultures were washed after exposure to TSH, the cellular cyclic AMP levels rapidly declined, suggesting that removal of receptor-bound TSH results in a prompt cessation of cyclic AMP production. The thyroid cells in monolayer also responded to cholera toxin. The response was dose-dependent, and cholera toxin as low as 1 ng/ml was able to increase cyclic AMP production. In contrast to the observations in TSH, the cyclic AMP responses induced by cholera were hardly affected by washing the cultures after exposure to cholera toxin. Treatment of the cells with cholera toxin for only 3 min resulted in a continuous stimulation of cyclic AMP production for more than 4 hours. Confirming recent observations by others, most of Graves' IgG stimulated cyclic AMP production in a dose-dependent manner, but some of them inhibited the response at high concentrations. IgG derived from normal subjects did not increase cellular cyclic AMP. The time course in the cyclic AMP responses induced by Graves' IgG was variable among the IgG preparations from different patients. In some patients, the maximal responses were attained after 4 hours of incubation. A significant difference was noted between TSH and Graves' IgG in the stimulation of cyclic AMP production after washing the cultures. When the cultures were treated with Graves' IgG for 30 min, washed and then incubated without Graves' IgG, cellular cyclic AMP levels remained at the levels which were almost equivalent to those observed in the continuous presence of the IgGs.(ABSTRACT TRUNCATED AT 400 WORDS)


[Giant sebaceous gland hyperplasia of the vulva].
Sebaceous gland hyperplasia is an epithelial tumour with sebaceous differentiation. Genital involvement is rare. In this paper, we report a new case of sebaceous gland hyperplasia of the vulva. A 27 year-old woman presented multiple polypoid lesions of the lower third of left labium majus. The lesions were soft to the touch, measured 5 cm in length and were painless. The cutaneous biopsy confirmed the diagnosis of sebaceous gland hyperplasia of the vulva. Surgical excision was performed in two separate procedures and was successful. This case was unusual in terms of the site, the clinical appearance and the weeping seen due to the high concentration of hyperplasic sebaceous glands.


A surface charge dependent enhanced Th1 antigen-specific immune response in lymph nodes by transfersome-based nanovaccine-loaded dissolving microneedle-assisted transdermal immunization.
The efficient delivery of vaccines to draining lymph nodes and the induction of robust local immune responses are crucial for immunotherapy. Transdermal administration has been evidenced to facilitate the delivery of ingredients to lymph nodes. In this study, transfersomes with opposite surface charges were applied for antigen encapsulation and these were integrated with dissolving microneedles to investigate their effects on immune responses via transdermal immunization. The microneedles were easily inserted into mouse skin and achieved the local release of nanovaccines into the dermis through dissolution. Although anionic nanovaccines promoted cellular uptake via DC2.4, cationic nanovaccines exhibited stronger escape capacities from endocytic compartments, facilitating antigen processing via an MHC-I presentation pathway, and formed larger accumulations in lymph nodes. Compared with their anionic counterparts, the cationic nanovaccines more efficiently activated DC maturation and induced Th1 immunity; this was suggested by the significantly increased IgG2a/IgG1 ratio and elevated cytokine secretion from Th1 cells, without an enhancement in the Th2 response. Such an enhanced Th1 antigen-specific immune response in lymph nodes via a transdermal vaccine delivery platform is beneficial for potential immunotherapy approaches.


Caterpillars selected for large body size and short development time are more susceptible to oxygen-related stress.
Recent studies suggest that higher growth rates may be associated with reduced capacities for stress tolerance and increased accumulated damage due to reactive oxygen species. We tested the response of Manduca sexta (Sphingidae) lines selected for large or small body size and short development time to hypoxia (10 kPa) and hyperoxia (25, 33, and 40 kPa); both hypoxia and hyperoxia reduce reproduction and oxygen levels over 33 kPa have been shown to increase oxidative damage in insects. Under normoxic (21 kPa) conditions, individuals from the large-selected (big-fast) line were larger and had faster growth rates, slightly longer developmental times, and reduced survival rates compared to individuals from a line selected for small size (small-fast) or an unselected control line. Individuals from the big-fast line exhibited greater negative responses to hyperoxia with greater reductions in juvenile and adult mass, growth rate, and survival than the other two lines. Hypoxia generally negatively affected survival and growth/size, but the lines responded similarly. These results are mostly consistent with the hypothesis that simultaneous acquisition of large body sizes and short development times leads to reduced capacities for coping with stressful conditions including oxidative damage. This result is of particular importance in that natural selection tends to decrease development time and increase body size.


Structure of human immunodeficiency virus type 1 Vpr(34-51) peptide in micelle containing aqueous solution.
Human immunodeficiency virus type 1 protein R (HIV-1 Vpr) promotes nuclear entry of viral nucleic acids in nondividing cells, causes G(2) cell cycle arrest and is involved in cellular differentiation and cell death. Vpr subcellular localization is as variable as its functions. It is known, that consistent with its role in nuclear transport, Vpr localizes to the nuclear envelope of human cells. Further, a reported ion channel activity of Vpr is clearly dependent on its localization in or at membranes. We focused our structural studies on the secondary structure of a peptide consisting of residues 34-51 of HIV-1 Vpr. This part of Vpr plays an important role in Vpr oligomerization, contributes to cell cycle arrest activity, and is essential for virion incorporation and binding to HHR23A, a protein involved in DNA repair. Employing NMR spectroscopy we found this part of Vpr to be almost completely alpha helical in the presence of micelles, as well as in trifluoroethanol containing and methanol/chloroform solvent. Our results provide structural data suggesting residues 34-51 of Vpr to contain an amphipathic, leucine-zipper-like alpha helix, which serves as a basis for oligomerization of Vpr and its interactions with cellular and viral factors involved in subcellular localization and virion incorporation of Vpr.


Older HIV+ and HIV- Adults Provide Similar Definitions of Successful Aging: A Mixed-Methods Examination.
As HIV-infected (HIV+) individuals age, there is a need to understand successful aging (SA) from the patient perspective. This study compared SA definitions between HIV+ and HIV-uninfected (HIV-) older adults and then examined correlates of SA categories. Ninety-three HIV+ and 46 HIV- older (aged 50+) adults provided brief definitions of SA, which was examined using content analysis. We then compared the frequency of SA categories by serostatus and examined the correlates of SA categories within both groups. Seven SA categories emerged: General Health, Cognitive Health & Ability, Physical/Biological Health & Ability, Social Relationships, Attitudes, Psychological, & Emotional Well-Being, Proactive & Engaged Lifestyle, and Independence. While no significant differences emerged, HIV- older adults were more likely to report General Health and the subcategory of Longevity/Survival, while HIV+ older adults were more likely to report subcategories of Enjoying Life & Fulfillment and Maintaining Balance. Few demographic correlates of SA categories emerged. Mood and HIV characteristics were not associated with SA categories. In both groups, those without neurocognitive impairment were significantly more likely to endorse General Health than those with neurocognitive impairment. HIV+ and HIV- older individuals may generally perceive SA similarly, and their definitions parallel with existing models of SA. Yet, living with a chronic illness may cause HIV+ older adults to place greater value on quality of life and life satisfaction than physical health and chronological age. Observational and intervention studies may use similar approaches in evaluating and maximizing SA.


HTLV-1 viral oncogene HBZ drives bone destruction in adult T cell leukemia.
Osteolytic bone lesions and hypercalcemia are common, serious complications in adult T cell leukemia/lymphoma (ATL), an aggressive T cell malignancy associated with human T cell leukemia virus type 1 (HTLV-1) infection. The HTLV-1 viral oncogene HBZ has been implicated in ATL tumorigenesis and bone loss. In this study, we evaluated the role of HBZ on ATL-associated bone destruction using HTLV-1 infection and disease progression mouse models. Humanized mice infected with HTLV-1 developed lymphoproliferative disease and continuous, progressive osteolytic bone lesions. HTLV-1 lacking HBZ displayed only modest delays to lymphoproliferative disease but significantly decreased disease-associated bone loss compared with HTLV-1-infected mice. Gene expression array of acute ATL patient samples demonstrated increased expression of RANKL, a critical regulator of osteoclasts. We found that HBZ regulated RANKL in a c-Fos-dependent manner. Treatment of HTLV-1-infected humanized mice with denosumab, a monoclonal antibody against human RANKL, alleviated bone loss. Using patient-derived xenografts from primary human ATL cells to induce lymphoproliferative disease, we also observed profound tumor-induced bone destruction and increased c-Fos and RANKL gene expression. Together, these data show the critical role of HBZ in driving ATL-associated bone loss through RANKL and identify denosumab as a potential treatment to prevent bone complications in ATL patients.


Increased genetic diversity of ADME genes in African Americans compared with their putative ancestral source populations and implications for pharmacogenomics.
African Americans have been treated as a representative population for African ancestry for many purposes, including pharmacogenomic studies. However, the contribution of European ancestry is expected to result in considerable differences in the genetic architecture of African American individuals compared with an African genome. In particular, the genetic admixture influences the genomic diversity of drug metabolism-related genes, and may cause high heterogeneity of drug responses in admixed populations such as African Americans. The genomic ancestry information of African-American (ASW) samples was obtained from data of the 1000 Genomes Project, and local ancestral components were also extracted for 32 core genes and 252 extended genes, which are associated with drug absorption, distribution, metabolism, and excretion (ADME) genes. As expected, the global genetic diversity pattern in ASW was determined by the contributions of its putative ancestral source populations, and the whole profiles of ADME genes in ASW are much closer to those in YRI than in CEU. However, we observed much higher diversity in some functionally important ADME genes in ASW than either CEU or YRI, which could be a result of either genetic drift or natural selection, and we identified some signatures of the latter. We analyzed the clinically relevant polymorphic alleles and haplotypes, and found that 28 functional mutations (including 3 missense, 3 splice, and 22 regulator sites) exhibited significantly higher differentiation between the three populations. Analysis of the genetic diversity of ADME genes showed differentiation between admixed population and its ancestral source populations. In particular, the different genetic diversity between ASW and YRI indicated that the ethnic differences in pharmacogenomic studies are broadly existed despite that African ancestry is dominant in Africans Americans. This study should advance our understanding of the genetic basis of the drug response heterogeneity between populations, especially in the case of population admixture, and have significant implications for evaluating potential inter-population heterogeneity in drug treatment effects.


Activation of TRPV1 channel by dietary capsaicin improves visceral fat remodeling through connexin43-mediated Ca2+ influx.
The prevalence of obesity has dramatically increased worldwide and has attracted rising attention, but the mechanism is still unclear. Previous studies revealed that transient receptor potential vanilloid 1 (TRPV1) channels take part in weight loss by enhancing intracellular Ca2+ levels. However, the potential mechanism of the effect of dietary capsaicin on obesity is not completely understood. Ca2+ transfer induced by connexin43 (Cx43) molecules between coupled cells takes part in adipocyte differentiation. Whether TRPV1-evoked alterations in Cx43-mediated adipocyte-to-adipocyte communication play a role in obesity is unknown. We investigated whether Cx43 participated in TRPV1-mediated adipocyte lipolysis in cultured 3T3-L1 preadipocytes and visceral adipose tissues from humans and wild-type (WT) and TRPV1-deficient (TRPV1-/-) mice. TRPV1 and Cx43 co-expressed in mesenteric adipose tissue. TRPV1 activation by capsaicin increased the influx of Ca2+ in 3T3-L1 preadipocytes and promoted cell lipolysis, as shown by Oil-red O staining. These effects were deficient when capsazepine, a TRPV1 antagonist, and 18 alpha-glycyrrhetinic acid (18α-GA), a gap-junction inhibitor, were administered. Long-term chronic dietary capsaicin reduced the weights of perirenal, mesenteric and testicular adipose tissues in WT mice fed a high-fat diet. Capsaicin increased the expression levels of p-CaM, Cx43, CaMKII, PPARδ and HSL in mesenteric adipose tissues from WT mice fed a high-fat diet, db/db mice, as well as obese humans, but these effects of capsaicin were absent in TRPV1-/- mice. Long-term chronic dietary capsaicin decreased the body weights and serum lipids of WT mice, but not TRPV1-/- mice, fed a high-fat diet. This study demonstrated that capsaicin activation of TRPV1-evoked increased Ca2+ influx in Cx43-mediated adipocyte-to-adipocyte communication promotes lipolysis in both vitro and vivo. TRPV1 activation by dietary capsaicin improves visceral fat remodeling through the up-regulation of Cx43.


Gap junction proteins in the light-damaged albino rat.
Changes in connexin expression are associated with many pathological conditions seen in animal models and in humans. We hypothesized that gap junctions are important mediators in tissue dysfunction and injury processes in the retina, and therefore, we investigated the pattern of connexin protein expression in the light-damaged albino rat eye. Adult Sprague-Dawley rats were exposed to intense light for 24 h. The animals were euthanized, and ocular tissue was harvested at 0 h, 6 h, 24 h, 48 h, and 7 days after light damage. The tissues were processed for immunohistochemistry and western blotting to analyze the expression of the gap junction proteins in the light-damaged condition compared to the non-light-damaged condition. Cell death was detected using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique. Intense light exposure caused increased TUNEL labeling of photoreceptor cells. Immunocytochemistry revealed that connexin 36 (Cx36) was significantly increased in the inner plexiform layer and Cx45 was significantly decreased in the light-damaged retina. The pattern of Cx36 and Cx45 labeling returned to normal 7 days after light damage. Cx43 significantly increased in the RPE and the choroid in the light-damaged tissue, and decreased but not significantly in the retina. This elevated Cx43 expression in the choroid colocalized with markers of nitration-related oxidative stress (nitrotyrosine) and inflammation (CD45 and ionized calcium-binding adaptor molecule-1) in the choroid. The results suggest that connexins are regulated differently in the retina than in the choroid in response to photoreceptor damage. Changes in connexins, including Cx36, Cx43, and Cx45, may contribute to the damage process. Specifically, Cx43 was associated with inflammatory damage. Therefore, connexins may be candidate targets for treatment for ameliorating disease progression.


[Carbohydrate metabolism in hypokalemic periodic paralysis (a clinico-histochemical study)].
The author presets some data on histochemical studies of muscle carbohydrate metabolism in patients with hypopotassemic periodic paralysis during the attack and in intermissions. The most constant and considerable changes were revealed during the paralytic attack and they lead to a decrease of phosphorylase activity, an increase of glycogen content and glucose decrease in the muscular fibers. In intermissions these changes were expressed either minimally or were absent altogether. The initial biochemical effect remains unknown. Undoubtedly, the carbohydrate metabolism is significantly damaged in this disease, and there is a definite parallelism between the severity of the clinical course and the degree of the revealed histochemical changes in the muscles.


An inaugural conference on occupational health in The Gambia: exploring the world through international occupational health programs.
This conference on workplace safety was a success for both the Gambians who attended and the Americans involved in planning, implementing, and funding the conference. The Americans were delighted to bring about some understanding of worker safety to a country that has struggled with labor issues for centuries. The dialogue reflected the concern that stakeholders in both countries have for labor issues, but there is a tremendous need for ongoing partnership with industrialized countries to bring these ideas and strategies to fruition. The Gambia has received some assistance in the area of economic development, but not in improving workplace safety. A wide range of economic support activities have been initiated by the ILO in The Gambia during the past 20 years. This has included providing guidance with international labor standards, assistance to the development of labor and related services, support to employer and worker organizations, vocational training, job creation and employment policy, and small enterprise development. Many challenges lie ahead for The Gambia in improving workplace safety. Serious concerns remain related to minimum safeguards in the workplace. For example, the Labor Act specifies safety equipment that an employer must provide for employees working in designated occupations, but these are rarely offered (Culp et al., 2003). Workers are plentiful and jobs are not, so workers tend to take any employment they can without regard to working conditions. The government has not formally revoked employers who have contributed to worker deaths. Industrial inspections are conducted, although there is little consequence for violations. The cycle of poverty, younger workers, and lack of education also contribute to many worker safety issues. Farm workers cannot read instructions on pesticide containers and industrial employees cannot understand workplace hazard communications. While the Gambian constitution mandates free compulsory primary education, the current state of educational opportunities is so dismal for poor families that many will not go beyond primary school and will enter the work force without technical skills. Teachers are overworked. While some young people could develop small business enterprises related to tourism and woodcarving, the vast majority continue a downward spiral into poverty (McGrath & King, 1999).


Is Total Disk Replacement a Cost-effective Treatment for Cervical Degenerative Disk Disease?
A retrospective study. The objective of this study was to assess the cost-effectiveness and clinical outcome of motion-preserving versus fusion procedures in cervical spine surgery. During the last decade there has been a huge growth in spine surgery with a concurrent increase in the economic burden. Currently, there appear to be no differences in clinical outcome between cervical total disk replacement (TDR) and spinal fusion (SF). For this reason it seems useful to know within the decision-making process whether there is a difference in actual cost between motion-preserving and fusion surgery. So far data that describe expenses involved in these procedures have not been available. This study offers a comparison of economic factors that should be considered in TDR and SF. The German statutory general healthcare insurance (GHI) provides anonymized patient-related data of their customers. A retrospective query using the codes of surgery of all TDR and SF surgery was performed from January 2003 to June 2008. A total of 467 cases with monosegmental or bisegmental surgery for degenerative disk pathologies were included. Both groups showed significant differences in independent variables such as age and sex (P<0.0001), but not in revision rates. Cost weight of diagnosis-related groups and length of hospitalization had a significant effect on total costs. Both groups obtained less pain medication postoperatively than preoperatively without a significant difference between each group. Postoperative absenteeism from work was significantly higher in the TDR group;however, patients with TDR underwent less rehabilitation covered by the GHI. Both groups had the same amount of preoperative and postoperative physiotherapy covered by the GHI. According to the collected data, there are no differences between the medical outcomes of cervical TDR in comparison with cervical SF. At the same time, while generating clinical results comparable with spinal fusion, TDR incurred significantly lower costs. Therefore, both from the medical and from the financial point of view, TDR is a viable choice in the treatment of degenerative disk pathology.


Attenuation of tick-borne encephalitis virus using large-scale random codon re-encoding.
Large-scale codon re-encoding (i.e. introduction of a large number of synonymous mutations) is a novel method of generating attenuated viruses. Here, it was applied to the pathogenic flavivirus, tick-borne encephalitis virus (TBEV) which causes febrile illness and encephalitis in humans in forested regions of Europe and Asia. Using an infectious clone of the Oshima 5-10 strain ("wild-type virus"), a cassette of 1.4kb located in the NS5 coding region, was modified by randomly introducing 273 synonymous mutations ("re-encoded virus"). Whilst the in cellulo replicative fitness of the re-encoded virus was only slightly reduced, the re-encoded virus displayed an attenuated phenotype in a laboratory mouse model of non-lethal encephalitis. Following intra-peritoneal inoculation of either 2.105 or 2.106 TCID50 of virus, the frequency of viraemia, neurovirulence (measured using weight loss and appearance of symptoms) and neuroinvasiveness (detection of virus in the brain) were significantly decreased when compared with the wild-type virus. Mice infected by wild-type or re-encoded viruses produced comparable amounts of neutralising antibodies and results of challenge experiments demonstrated that mice previously infected with the re-encoded virus were protected against subsequent infection by the wild-type virus. This constitutes evidence that a mammalian species can be protected against infection by a virulent wild-type positive-stranded RNA virus following immunisation with a derived randomly re-encoded strain. Our results demonstrate that random codon re-encoding is potentially a simple and effective method of generating live-attenuated vaccine candidates against pathogenic flaviviruses.


Bracket Placement According to Malocclusion.
This article reports the orthodontic treatment ofa 25-year-old female patient whose chief complaint was the inclination of the maxillary occlusal plane in front view. The individualized vertical placement of brackets is described. This placement made possible a symmetrical occlusal plane to be achieved in a rather straightforward manner without the need for further technical resources.


Australia's first transition to professional practice in primary care program for graduate registered nurses: a pilot study.
Increases in ageing, chronic illness and complex co-morbidities in the Australian population are adding pressure to the primary care nursing workforce. Initiatives to attract and retain nurses are needed to establish a sustainable and skilled future primary care nursing workforce. We implemented a transition to professional practice program in general practice settings for graduate nurses and evaluated graduate nurse competency, the graduate nurse experience and program satisfaction. This study aimed to determine whether a transition to professional practice program implemented in the general practice setting led to competent practice nurses in their first year post-graduation. A longitudinal, exploratory mixed-methods design was used to assess the pilot study. Data were collected at three times points (3, 6, 12 months) with complete data sets from graduate nurses (n = 4) and preceptors (n = 7). We assessed perceptions of the graduates' nursing competency and confidence, satisfaction with the preceptor/graduate relationship, and experiences and satisfaction with the program. Graduate nurse competency was assessed using the National Competency Standards for Nurses in General Practice. Semi-structured interviews with participants at Time 3 sought information about barriers, enablers, and the perceived impact of the program. Graduate nurses were found to be competent within their first year of clinical practice. Program perceptions from graduate nurses and preceptors were positive and the relationship between the graduate nurse and preceptor was key to this development. With appropriate support registered nurses can transition directly into primary care and are competent in their first year post-graduation. While wider implementation and research is needed, findings from this study demonstrate the potential value of transition to professional practice programs within primary care as a nursing workforce development strategy.


The Resazurin Reduction Assay Can Distinguish Cytotoxic from Cytostatic Compounds in Spheroid Screening Assays.
Spheroid-based cellular screening approaches represent a highly physiologic experimental setup to identify novel anticancer drugs and an innovative preclinical model to reduce the high failure rate of anticancer compounds in clinical trials. The resazurin reduction (RR) assay, known as the alamarBlue or CellTiter-Blue assay, is frequently used to determine cell viability/proliferation capacity in eukaryotic cells. Whether this assay is applicable to assess viability in multicellular spheroids has not been evaluated. We analyzed the RR assay to measure cytotoxic and/or cytostatic responses in tumor cell spheroids compared with conventional 2D cultures. We found that tight cell-cell interactions in compact spheroids hamper resazurin uptake and its subsequent reduction to resorufin, leading to lowered reduction activity in relation to the actual cellular health/cell number. Treatment with staurosporine disrupted close cell-cell contacts, which increased resazurin reduction compared with untreated controls. Loss of tight junctions by trypsinization or addition of EGTA or EDTA restored high resazurin reduction rates in untreated spheroids. In conclusion, the RR assay is unsuited to quantitatively measure cellular health/cell number in compact spheroids. However, it can be used to distinguish between cytotoxic versus cytostatic compounds in spheroids. Restoration of the correlation of cell viability/number to resazurin reduction capacity can be achieved by disruption of tight junctions.


Personalizing adjuvant therapy for patients with colorectal cancer.
The current standard-of-care adjuvant treatment for patients with colorectal cancer (CRC) comprises a fluoropyrimidine (5-fluorouracil or capecitabine) as a single agent or in combination with oxaliplatin, for either 3 or 6 months. Selection of therapy depends on conventional histopathological staging procedures, which constitute a blunt tool for patient stratification. Given the relatively marginal survival benefits that patients can derive from adjuvant treatment, improving the safety of chemotherapy regimens and identifying patients most likely to benefit from them is an area of unmet need. Patient stratification should enable distinguishing those at low risk of recurrence and a high chance of cure by surgery from those at higher risk of recurrence who would derive greater absolute benefits from chemotherapy. To this end, genetic analyses have led to the discovery of germline determinants of toxicity from fluoropyrimidines, the identification of patients at high risk of life-threatening toxicity, and enabling dose modulation to improve safety. Thus far, results from analyses of resected tissue to identify mutational or transcriptomic signatures with value as prognostic biomarkers have been rather disappointing. In the past few years, the application of artificial intelligence-driven models to digital images of resected tissue has identified potentially useful algorithms that stratify patients into distinct prognostic groups. Similarly, liquid biopsy approaches involving measurements of circulating tumour DNA after surgery are additionally useful tools to identify patients at high and low risk of tumour recurrence. In this Perspective, we provide an overview of the current landscape of adjuvant therapy for patients with CRC and discuss how new technologies will enable better personalization of therapy in this setting.


Michelin tire baby syndrome--a case report and literature review.
Michelin tire syndrome is described in a two month old infant of Filipino-Saudi parents. The infant had generalized excessive folding of skin and facial dysmorphism. The skin biopsy showed excessive adipose tissue in reticular dermis, papillary dermis and around adnexa. Spontaneous partial improvement in skin folding was noted on follow up. To the best of our knowledge, this is the first case ever reported locally of Michelin tire.


Orthostatic headaches in the syndrome of the trephined: resolution following cranioplasty.
To draw attention to the syndrome of the trephined as a potential cause for orthostatic headaches without cerebrospinal fluid (CSF) leak. Orthostatic headaches typically result from CSF leaks but sometimes may occur in conditions without any evidence of CSF leakage. A 37-year-old right-handed woman became comatose after a motor vehicle accident with cerebral contusions and massive left cerebral edema. A large frontoparietal craniectomy was carried out. In 5 months, she made good neurologic recovery. Freeze-preserved bone flap was placed back. In several weeks she was functionally near normal. Two years later, she began to complain of orthostatic headache and gradually additional manifestations appeared including progressive gait unsteadiness, imprecise speech, cognitive difficulties, and an increasing left hemiparesis along with progressive sinking of the skull defect and shift of the midline and ventricular distortion. She underwent removal of resorptive sinking bone flap and construction of an acrylic cranioplasty. At 6-month follow-up, there was complete resolution of the orthostatic headaches, remarkable neurologic improvement along with resolution of midline shift and ventricular distortion. The syndrome of the trephined is yet another cause of orthostatic headaches without CSF leak.


Scope and limitations of the double [4+3]-cycloadditions of 2-oxyallyl cations to 2,2'-methylenedifuran and derivatives.
The reactivity of various 2-oxyallyl cations toward 2,2'-methylenedifuran (1b), 2,2'-(hydroxymethyl)difuran (1c), 2,2'-(trimethylsilylmethylene)difuran (1d), and di(2-furyl)methanone (1e) has been explored. Difuryl derivatives 1c, 1d, and 1e refused to undergo formal double [4+3]-cycloadditions. Conditions have been found to convert 1b into meso-1,1'-methylenedi[(1R,1'S,5S,5'R)- (3) and (+/-)-1,1'-methylenedi[(1RS,1'SR,5SR,5'RS)-8-oxabicyclo[3.2.1]oct-6-en-3-one] (4) that do not require CF(3)CH(OH)CF(3) as solvent. High yields of meso-1,1'-methylenedi[(1R,1'S,2S,2'R,4R,4'S,5S,5'R)- (5) and (+/-)-1,1'-methylenedi[(1RS,1'RS,2SR,2'SR,4RS,4'RS,5SR,5'SR)-2,4-dimethyl-8-oxabicyclo[3.2.1]oct-6-en-3-one] (6) have been obtained when 1b was reacted with 2,4-dibromopentan-3-one (7h) and NaI/Cu.


[Effects of PGF2 alpha prostaglandins on milk ejection in the ewe. Consequences of ovariectomy with or without estroprogestative complementation].
The intrajugular administration of 200 micrograms of Dinolytic (tromethamine salt of PGF2 alpha) to lactating ewes caused an intramammary pressure (IMP) increase only during the luteal phase of the sexual cycle (group A). The increase in progesteronemia, induced by three daily injections of 2.5 micrograms of LH-RH, did not modify response amplitude (group B). On the other hand, bilateral ovariectomy (groupe C) led to the suppression of those responses, but supplementary oestroprogestative (table 1) did not re-establish them. It is probable that the milk ejection caused by PGF2 alpha resulted from a release of oxytocin by the corpus luteum. The ineffectiveness of the exogenous and endogenous reinforcement of progesterone therefore suggests that this hormone plays no part in the putative control of hypothalamic receptivity to prostaglandins.


Chromosome 4q25 variants and biomarkers of myocardial fibrosis in patients with atrial fibrillation.
Little is known about how genetic predisposition and fibrosis relate in atrial fibrillation (AF). Hence, we sought to determine whether the genetic variants and biomarkers for fibrosis enhance prediction of outcomes after catheter ablation. Consecutive patients who underwent catheter ablation of AF (paroxysmal, 158; nonparoxysmal, 137) or supraventricular tachycardia without AF (n = 70) were studied retrospectively. Plasma levels of transforming growth factor β1 (TGF-β1), tissue inhibitor of metalloproteinase 1 (TIMP-1), and 4q25 single-nucleotide polymorphisms (SNPs) (rs10033464 and rs220073) were measured. Mean plasma levels of both TGF-β1 and TIMP-1 were higher in patients with AF than in the control (all P < .001). Plasma levels of TIMP-1 were higher in patients with recurrence compared with those without recurrence (P = .039). Patients with variant alleles of rs10033464 showed increased recurrence after catheter ablation in patients with paroxysmal AF including after adjustment (P = .027). Patients with TIMP-1 < 107 ng/mL and no variant allele (GG) at rs10033464 had lower recurrence rates compared with other groups in those with paroxysmal AF (logrank; P = .007), whereas there was no significant difference among those patients with persistent forms of AF. Inclusion of biomarkers and genotype improved discrimination of AF recurrence in patients with paroxysmal AF (C-statistic .499 vs .600). The combination of plasma TIMP-1 concentrations less than 107 ng/mL and the absence of a variant allele at rs10033464 was associated with lower recurrence rates in patients with paroxysmal AF. This study suggests that 4q25 SNPs and biomarkers for fibrosis may provide additive value in risk stratification for AF recurrence after catheter ablation.


Differential induction of Ly6G and Ly6C positive myeloid derived suppressor cells in chronic kidney and liver inflammation and fibrosis.
CD11b+Gr1+ myeloid derived suppressor cells (MDSC) are known to be very potent suppressors of T cell immunity and can be further stratified into granulocytic MDSC and monocytic MDSC in mice based on expression of Ly6G or Ly6C, respectively. Here, using these markers and functional assays, we aimed to identify whether MDSC are induced during chronic inflammation leading to fibrosis in both kidney and liver and whether additional markers could more specifically identify these MDSC subsets. In an adenine-induced model of kidney inflammation/fibrosis suppressive Ly6Gpos MDSC were induced. The suppressive function within the Ly6G+ MDSC population was exclusively present in IFNγRβ expressing cells. In contrast, in chronic inflammation in the liver induced by bile duct ligation, suppressive capacity was exclusively present in the Ly6Cpos MDSC subset. Gene expression analyses confirmed the differential origins and regulation of those MDSC subsets. Additionally, depletion of MDSC in either kidney or liver fibrosis enhanced fibrosis markers, indicating a protective role for MDSC in organ fibrosis. Thus, our data demonstrate that during liver inflammation and kidney fibrosis MDSC with similar function arise bearing a distinct marker profile and arising from different cell populations.


Neural stem cells: a potential source for remyelination in neuroinflammatory disease.
In multiple sclerosis, the central nervous system is lesioned through invasion of plaque-forming inflammatory cells, primarily contributing to immune attack of myelin and oligodendrocytes. In this report we address the possible activation and differentiation of central nervous system stem cells following such immunological insults in a well-characterized rat model of multiple sclerosis characterised by spinal cord pathology. Dye-labeled central nervous system stem cells, residing within the ependymal layer of the central canal responded to the multiple sclerosis-like conditions by proliferation, while some of the migrating stem cell-derived cells expressed markers typical for oligodendrocytes (04) and astrocytes (glial fibrillary acidic protein, GFAP) in the demyelinated area. Our results indicate that regenerative stem cell activation following immunoactivity is different from that after trauma, exemplified by the slower time course of stem cell proliferation and migration of progeny, in addition to the ability of the stem cell-derived cells to express oligodendrocyte markers. Finally, deleterious effects of macrophages on the stem cell population were evident and may contribute to the depletion of the stem cell population in neuroinflammatory disorders.


COVID-19 and oral diseases: Assessing manifestations of a new pathogen in oral infections.
Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is a recently identified virus responsible for life-threatening coronavirus disease 19 (COVID-19). The SARS-CoV-2 infected subjects can be asymptomatic or symptomatic; the later may present a wide spectrum of clinical manifestations. However, the impact of SARS-CoV-2 on oral diseases remain poorly studied. Detection of SARS-CoV-2 in saliva indicates existence of virus in the oral cavity. Recent studies demonstrating the expression of ACE-2, a SARS-CoV-2 entry receptor, in oral tissues further strengthens this observation. Cytokine storm in severe COVID-19 patients and copious secretion of pro-inflammatory cytokines (IL-6, IL-1β and TNF-α) in multiple symptomatic oral pathologies including periodontitis and periapical periodontitis suggests that inflammatory microenvironment is a hallmark of both COVID-19 and oral diseases. Hyperinflammation may provide conducive microenvironment for the growth of local oral pathogens or opportunistic microbes and exert detrimental impact on the oral tissue integrity. Multiple case reports have indicated uncharacterized oral lesions, symptomatic irreversible pulpitis, higher plaque index, necrotizing/desquamative gingivitis in COVID-19 patients suggesting that SARS-CoV-2 may worsen the manifestations of oral infections. However, the underlying factors and pathways remain elusive. Here we summarize current literature and suggest mechanisms for viral pathogenesis of oral dental pathology derived from oral microbiome and oral mucosa-dental tissue interactions. Longitudinal studies will reveal how the virus impairs disease progression and resolution post-therapy. Some relationships we suggest provide the basis for novel monitoring and treatment of oral viral disease in the era of SARS-CoV-2 pandemic, promoting evidence-based dentistry guidelines to diagnose virus-infected patients to improve oral health.


Phase 2, multicenter GIBB study of obinutuzumab plus bendamustine in previously untreated patients with chronic lymphocytic leukemia.
The single-arm, multicenter, phase 2 GIBB study (NCT02320487) investigated bendamustine plus obinutuzumab (BG) in previously untreated CLL. Patients (N = 102) received six cycles of intravenous obinutuzumab (cycle [C] 1: 100 mg day 1/900 mg day 2, and 1000 mg days 8/15; C2-6 1000 mg day 1) plus bendamustine (C1 90 mg/m2 days 2/3; C2-6 days 1/2). Complete response (CR), the primary endpoint, was 50%, overall response 89%. Estimated 2-year progression-free survival (PFS) and overall survival (OS) were 86% and 97%, respectively. Following initial minimal residual disease (MRD) negativity, median MRD negativity duration was 28.9 months. Undetectable MRD (<10-4) was observed in up to 83% of evaluable patients in peripheral blood (any time) and 59% in bone marrow at response evaluation. Most common grade 3/4 adverse events (AEs) were neutropenia (25%; 5% febrile) and infusion-related reactions (9%). BG proved clinically active in CLL with high response, MRD negativity, and survival rates, consistent with other first-line studies of anti-CD20 antibody/bendamustine combinations.


A pocket guide on how to structure SARS-CoV-2 drugs and therapies.
The race to identify a successful treatment for COVID19 will be defined by fundamental research into the replication cycle of the SARS-CoV-2 virus. This has identified five distinct stages from which numerous vaccination and clinical trials have emerged alongside an innumerable number of drug discovery studies currently in development for disease intervention. Informing every step of the viral replication cycle has been an unprecedented 'call-to-arms' by the global structural biology community. Of the 20 main SARS-CoV-2 proteins, 13 have been resolved structurally for SARS-CoV-2 with most having a related SARS-CoV and MERS-CoV structural homologue totalling some 300 structures currently available in public repositories. Herein, we review the contribution of structural studies to our understanding of the virus and their role in structure-based development of therapeutics.


The spectrum of levodopa-induced dyskinesias.
The most common types of levodopa-induced dyskinesias in patients with Parkinson's disease (PD) are chorea and dystonia, and often the two types are intermixed. Myoclonus is a far less common problem. The dyskinesias tend to develop over time, not usually being encountered at the initiation of levodopa therapy. Eventually, they affect more than 50% of patients on long-term levodopa treatment. Once they appear, they are difficult to eliminate. Substituting weaker dopaminergic agents for levodopa often fails to eliminate the dyskinesias. Most of the dyskinesias occur at the time of the highest brain concentration of levodopa and its product, dopamine--so-called peak-dose dyskinesias. Chorea and dystonia, usually in the legs, occur less commonly at the beginning and end of dosing, and these are called diphasic dyskinesias. Dystonia can also occur during the 'off' state, i.e. when the levodopa concentration is low. These 'off' dystonias are often painful and must be distinguished from peak-dose dystonia and from dystonia that may be a feature of PD itself.


Effectiveness evaluation of glyphosate oxidation employing the H(2)O(2)/UVC process: toxicity assays with Vibrio fischeri and Rhinella arenarum tadpoles.
The H(2)O(2)/UVC process was applied to the photodegradation of a commercial formulation of glyphosate in water. Two organisms (Vibrio fischeri bacteria and Rhinella arenarum tadpoles) were used to investigate the toxicity of glyphosate in samples M(1,) M(2), and M(3) following different photodegradation reaction times (120, 240 and 360 min, respectively) that had differing amounts of residual H(2)O(2). Subsamples of M(1), M(2), and M(3) were then used to create samples M(1,E), M(2,E) and M(3,E) in which the H(2)O(2) had been removed. Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities were measured in tadpoles to determine possible sub-lethal effects. In V. fischeri, M(1,E), which was collected early in the photodegradation process, caused 52% inhibition, while M(3,E), which was collected at the end of the photodegradation process, caused only 17% inhibition. Survival of tadpoles was 100% in samples M(2), M(3), and in M(1,E), M(2,E) and M(3,E). The lowest percentages of enzymatic inhibition were observed in samples without removal of H(2)O(2): 13.96% (AChE) and 16% (BChE) for M(2), and 24.12% (AChE) and 13.83% (BChE) for M(3). These results show the efficiency of the H(2)O(2)/UVC process in reducing the toxicity of water or wastewater polluted by commercial formulations of glyphosate. According to the ecotoxicity assays, the conditions corresponding to M(2) (11 ± 1 mg a.e. L(-1) glyphosate and 11 ± 1 mg L(-1) H(2)O(2)) could be used as a final point for glyphosate treatment with the H(2)O(2)/UV process.


RIPiT-Seq: a high-throughput approach for footprinting RNA:protein complexes.
Development of high-throughput approaches to map the RNA interaction sites of individual RNA binding proteins (RBPs) transcriptome-wide is rapidly transforming our understanding of post-transcriptional gene regulatory mechanisms. Here we describe a ribonucleoprotein (RNP) footprinting approach we recently developed for identifying occupancy sites of both individual RBPs and multi-subunit RNP complexes. RNA:protein immunoprecipitation in tandem (RIPiT) yields highly specific RNA footprints of cellular RNPs isolated via two sequential purifications; the resulting RNA footprints can then be identified by high-throughput sequencing (Seq). RIPiT-Seq is broadly applicable to all RBPs regardless of their RNA binding mode and thus provides a means to map the RNA binding sites of RBPs with poor inherent ultraviolet (UV) crosslinkability. Further, among current high-throughput approaches, RIPiT has the unique capacity to differentiate binding sites of RNPs with overlapping protein composition. It is therefore particularly suited for studying dynamic RNP assemblages whose composition evolves as gene expression proceeds.


Partners in caregiving in a special care environment: cooperative communication between staff and families on dementia units.
This article reports the results of a randomized, controlled evaluation of Partners in Caregiving in a Special Care Environment, an intervention designed to improve communication and cooperation between staff and families of residents in nursing home dementia programs. Participants included 388 family members and 384 nursing staff members recruited from 20 nursing homes, randomly assigned to treatment and control conditions. Project staff conducted training sessions on communication and conflict-resolution techniques with two groups at the intervention sites: staff and residents' family members, followed by a joint meeting with facility administrators. Families, staff, residents, and facility programs in the intervention facilities all demonstrated positive outcomes from program participation. Families experienced significant improvement in communicating with staff and in staff behaviors toward them, and spouses of residents increased their care involvement. Staff reported reduced conflict with families and reduced depression; burnout for nurses increased for individuals in the control group but not those in the treatment group. Behavioral symptoms decreased for residents, and facilities implemented more family-focused programs. Effective staff and family partnerships are critical in caring for residents with dementia. The Partners in Caregiving in a Special Care Environment program is an evidence-based intervention that enables these partnerships to develop and thrive, translating into improved experiences for residents, families, and staff.


HIV risk behaviors among inner-city African American women. The Community Housing AIDS Prevention Study Group.
This study examined the prevalence and predictors of HIV risk behaviors among a sample of 875 low-income, African American women residents of inner-city housing developments. The women completed an anonymous questionnaire that revealed that one third of them were at high risk for HIV either because they had multiple partners or because of the high-risk behaviors of their regular partner. HIV risk was highest among women who accurately perceived themselves to be at increased HIV risk, reported weak behavioral intentions to reduce risk, and held stronger beliefs about psychosocial barriers to condom use. Women at high risk were also younger, reported higher rates of substance use, and indicated that their housing development lacked social cohesiveness. These findings suggest that HIV prevention efforts for this population should focus on strengthening women's risk reduction behavioral intentions and self-efficacy through skill development, overcoming psychosocial barriers to condom use, managing the risk related to substance use, and incorporating approaches that take into account the social, psychological, and relationship barriers to change among economically impoverished African American women.