What confidence should we have in GRADE?
Confidence (or belief) that a therapy is effective is essential to practicing clinical medicine. GRADE, a popular framework for developing clinical recommendations, provides a means for assigning how much confidence one should have in a therapy's effect estimate. One's level of confidence (or "degree of belief") can also be modelled using Bayes theorem. In this paper, we look through both a GRADE and Bayesian lens to examine how one determines confidence in the effect estimate. Philosophical examination. The GRADE framework uses a criteria-based method to assign a quality of evidence level. The criteria pertain mostly to considerations of methodological rigour, derived from a modified evidence-based medicine evidence hierarchy. The four levels of quality relate to the level of confidence one should have in the effect estimate. The Bayesian framework is not bound by a predetermined set of criteria. Bayes theorem shows how a rational agent adjusts confidence (ie, degree of belief) in the effect estimate on the basis of the available evidence. Such adjustments relate to the principles of incremental confirmation and evidence proportionism. Use of the Bayesian framework reveals some potential pitfalls in GRADE's criteria-based thinking on confidence that are out of step with our intuitions on evidence. A rational thinker uses all available evidence to formulate beliefs. The GRADE criteria seem to suggest that we discard some of that information when other, more favoured information (eg, derived from clinical trials) is available. The GRADE framework should strive to ensure that the whole evidence base is considered when determining confidence in the effect estimate. The incremental value of such evidence on determining confidence in the effect estimate should be assigned in a manner that is theoretically or empirically justified, such that confidence is proportional to the evidence, both for and against it.


An interdisciplinary heuristic evaluation method for universal building design.
This study highlights how heuristic evaluation as a usability evaluation method can feed into current building design practice to conform to universal design principles. It provides a definition of universal usability that is applicable to an architectural design context. It takes the seven universal design principles as a set of heuristics and applies an iterative sequence of heuristic evaluation in a shopping mall, aiming to achieve a cost-effective evaluation process. The evaluation was composed of three consecutive sessions. First, five evaluators from different professions were interviewed regarding the construction drawings in terms of universal design principles. Then, each evaluator was asked to perform the predefined task scenarios. In subsequent interviews, the evaluators were asked to re-analyze the construction drawings. The results showed that heuristic evaluation could successfully integrate universal usability into current building design practice in two ways: (i) it promoted an iterative evaluation process combined with multi-sessions rather than relying on one evaluator and on one evaluation session to find the maximum number of usability problems, and (ii) it highlighted the necessity of an interdisciplinary ad hoc committee regarding the heuristic abilities of each profession. A multi-session and interdisciplinary heuristic evaluation method can save both the project budget and the required time, while ensuring a reduced error rate for the universal usage of the built environments.


Understanding Detrimental and Beneficial Grain Boundary Effects in Halide Perovskites.
Grain boundaries play a key role in the performance of thin-film optoelectronic devices and yet their effect in halide perovskite materials is still not understood. The biggest factor limiting progress is the inability to identify grain boundaries. Noncrystallographic techniques can misidentify grain boundaries, leading to conflicting literature reports about their influence; however, the gold standard - electron backscatter diffraction (EBSD) - destroys halide perovskite thin films. Here, this problem is solved by using a solid-state EBSD detector with 6000 times higher sensitivity than the traditional phosphor screen and camera. Correlating true grain size with photoluminescence lifetime, carrier diffusion length, and mobility shows that grain boundaries are not benign but have a recombination velocity of 1670 cm s-1 , comparable to that of crystalline silicon. Amorphous grain boundaries are also observed that give rise to locally brighter photoluminescence intensity and longer lifetimes. This anomalous grain boundary character offers a possible explanation for the mysteriously long lifetime and record efficiency achieved in small grain halide perovskite thin films. It also suggests a new approach for passivating grain boundaries, independent of surface passivation, to lead to even better performance in optoelectronic devices.


Endocytosis‒Mediated Invasion and Pathogenicity of Streptococcus agalactiae in Rat Cardiomyocyte (H9C2).
Streptococcus agalactiae infection causes high mortality in cardiovascular disease (CVD) patients, especially in case of setting prosthetic valve during cardiac surgery. However, the pathogenesis mechanism of S. agalactiae associate with CVD has not been well studied. Here, we have demonstrated the pathogenicity of S. agalactiae in rat cardiomyocytes (H9C2). Interestingly, both live and dead cells of S. agalactiae were uptaken by H9C2 cells. To further dissect the process of S. agalactiae internalization, we chemically inhibited discrete parts of cellular uptake system in H9C2 cells using genistein, chlorpromazine, nocodazole and cytochalasin B. Chemical inhibition of microtubule and actin formation by nocodazole and cytochalasin B impaired S. agalactiae internalization into H9C2 cells. Consistently, reverse‒ transcription PCR (RT‒PCR) and quantitative real time‒PCR (RT-qPCR) analyses also detected higher levels of transcripts for cytoskeleton forming genes, Acta1 and Tubb5 in S. agalactiae‒infected H9C2 cells, suggesting the requirement of functional cytoskeleton in pathogenesis. Host survival assay demonstrated that S. agalactiae internalization induced cytotoxicity in H9C2 cells. S. agalactiae cells grown with benzyl penicillin reduced its ability to internalize and induce cytotoxicity in H9C2 cells, which could be attributed with the removal of surface lipoteichoic acid (LTA) from S. agalactiae. Further, the LTA extracted from S. agalactiae also exhibited dose‒dependent cytotoxicity in H9C2 cells. Taken together, our data suggest that S. agalactiae cells internalized H9C2 cells through energy‒dependent endocytic processes and the LTA of S. agalactiae play major role in host cell internalization and cytotoxicity induction.


Identification and characterization of IS1381, a new insertion sequence in Streptococcus pneumoniae.
A new insertion sequence (IS1381) was identified in the genome of Streptococcus pneumoniae R6 as an 846-bp segment containing 20-bp terminal inverted repeats and flanked by 7-bp direct repeats. The three sequenced copies of this element have two overlapping open reading frame (ORF) genes named orfA and orfB. However, significant variations between individual copies were found, suggesting that inactivating mutations have occurred in an original single ORF. Accordingly, the consensus IS1381 element derived from the comparison of the three available copies should contain a single ORF sufficient to encode a basic protein of 267 amino acids which exhibited high similarity to the putative transposases of ISL2 from Lactobacillus helveticus and of IS702 from the cyanobacterium Calothrix sp. strain PCC 7601. A minimum of five to seven copies were detected by hybridization experiments in the R6 genome. In remarkable contrast with the two previously reported pneumococcal insertion sequences, several copies of IS1381 have been detected in all of the clinical isolates tested so far. Interestingly, Streptococcus oralis NCTC 11427 (type strain), a close relative of pneumococcus, does not contain this element, but its occurrence in the type strain of Streptococcus mitis (NCTC 12261) suggests that this species has exchanged DNA with S. pneumoniae directly or through an intermediate species.


Clinical safety of England's national programme for IT: a retrospective analysis of all reported safety events 2005 to 2011.
To analyse patient safety events associated with England's national programme for IT (NPfIT). Retrospective analysis of all safety events managed by a dedicated IT safety team between September 2005 and November 2011 was undertaken. Events were reviewed against an existing classification for problems associated with IT. The proportion of reported events per problem type, consequences, source of report, resolution within 24h, time of day and day of week were examined. Sub-group analyses were undertaken for events involving patient harm and those that occurred on a large scale. Of the 850 events analysed, 68% (n=574) described potentially hazardous circumstances, 24% (n=205) had an observable impact on care delivery, 4% (n=36) were a near miss, and 3% (n=22) were associated with patient harm, including three deaths (0·35%). Eleven events did not have a noticeable consequence (1%) and two were complaints (<1%). Amongst the events 1606 separate contributing problems were identified. Of these 92% were predominately associated with technical rather than human factors. Problems involving human factors were four times as likely to result in patient harm than technical problems (25% versus 8%; OR 3·98, 95%CI 1·90-8.34). Large-scale events affecting 10 or more individuals or multiple IT systems accounted for 23% (n=191) of the sample and were significantly more likely to result in a near miss (6% versus 4%) or impact the delivery of care (39% versus 20%; p<0·001). Events associated with NPfIT reinforce that the use of IT does create hazardous circumstances and can lead to patient harm or death. Large-scale patient safety events have the potential to affect many patients and clinicians, and this suggests that addressing them should be a priority for all major IT implementations.


Linear- and circular-plasmid copy numbers in Borrelia burgdorferi.
Borrelia burgdorferi, the Lyme disease agent, and other members of the spirochetal genus Borrelia have double-stranded linear plasmids in addition to supercoiled circular plasmids. The copy number relative to the chromosome was determined for 49- and 16-kb linear plasmids and a 27-kb circular plasmid of the type strain, B31, of B. burgdorferi. All three plasmids were present in low copy number, about one per chromosome equivalent, as determined by relative hybridizations of replicon-specific DNA probes. The low copy number of Borrelia plasmids suggests that initiation of DNA replication and partitioning are carefully controlled during the cell division cycle. The copy numbers of these three plasmids of strain B31 were unchanged after approximately 7,000 generations in continuous in vitro culture. A clone of B. burgdorferi B31 that did not contain the 16-kb linear plasmid was obtained after exposure of a culture to novobiocin, a DNA gyrase inhibitor. The plasmid-cured strain contains only one linear plasmid, the 49-kb plasmid, and thus has the smallest genome reported to date for B. burgdorferi.


The independent and interactive effects of navicular drop and quadriceps angle on neuromuscular responses to a weight-bearing perturbation.
Little is known about the effects of static alignment on neuromuscular control of the knee during dynamic motion. To evaluate the isolated and combined effects of quadriceps angle (QA) and navicular drop (ND) on neuromuscular responses to a weight-bearing perturbation. Mixed-model, repeated-measures design. Sports medicine and athletic training research laboratory. Seventy-nine National Collegiate Athletic Association Division I collegiate female athletes, classified with below-average ND and QA (LND-LQA); below-average ND and above-average QA (LND-HQA); above-average ND and below-average QA (HND-LQA); or above-average ND and QA (HND-HQA). A lower extremity perturbation device produced a forward and either internal or external rotation of the trunk and femur on the weight-bearing tibia to evoke a reflex response. Neuromuscular responses were examined in the quadriceps, hamstrings, and gastrocnemius muscles: preperturbation amplitude 50 milliseconds before the perturbation, reflex time, and postperturbation amplitude 150 milliseconds immediately postperturbation. Navicular drop had the greatest effect on preperturbation amplitude of the lateral hamstrings and postperturbation amplitude of all muscles, with greater activation amplitude noted in subjects in the HND classifications. Quadriceps angle primarily affected reflex time of the quadriceps; in subjects with LQA, reflex time was faster for internal rotation than external rotation perturbations. The interaction between ND and QA had the greatest effect on reflex time of the lateral hamstrings. For internal rotation perturbations, subjects in the LND classifications had faster reflex times in the lateral hamstrings if they had HQA values rather than LQA values. With external rotation perturbations, HND-LQA subjects had slower reflex times than those in all other alignment classifications. Navicular drop and QA have both independent and interactive effects on neuromuscular responses to a weight-bearing, rotational perturbation. These interactive effects highlight the importance of considering the entire lower extremity posture rather than a single alignment characteristic, given the potential for one alignment factor to compensate for or interact with another.


Dual muscle-liver transduction imposes immune tolerance for muscle transgene engraftment despite preexisting immunity.
Immune responses to therapeutic transgenes are a potential hurdle to treat monogenic muscle disorders. These responses result from the neutralizing activity of transgene-specific B cells and cytotoxic T cells recruited upon gene transfer. We explored here how dual muscle-liver expression of a foreign transgene allows muscle transgene engraftment after adenoassociated viral vector delivery. We found in particular that induction of transgene-specific tolerance is imposed by concurrent muscle and liver targeting, resulting in the absence of CD8+ T cell responses to the transgene. This tolerance can be temporally decoupled, because transgene engraftment can be achieved in muscle weeks after liver transduction. Importantly, transgene-specific CD8+ T cell tolerance can be established despite preexisting immunity to the transgene. Whenever preexisting, transgene-specific CD4+ and CD8+ memory T cell responses are present, dual muscle-liver transduction turns polyclonal, transgene-specific CD8+ T cells into typically exhausted T cells with high programmed cell death 1 (PD-1) expression and lack of IFN-γ production. Our results demonstrate that successful transduction of muscle tissue can be achieved through liver-mediated control of humoral and cytotoxic T cell responses, even in the presence of preexisting immunity to the muscle-associated transgene.


Stormwater runoff drives viral community composition changes in inland freshwaters.
Storm events impact freshwater microbial communities by transporting terrestrial viruses and other microbes to freshwater systems, and by potentially resuspending microbes from bottom sediments. The magnitude of these impacts on freshwater ecosystems is unknown and largely unexplored. Field studies carried out at two discrete sites in coastal Virginia (USA) were used to characterize the viral load carried by runoff and to test the hypothesis that terrestrial viruses introduced through stormwater runoff change the composition of freshwater microbial communities. Field data gathered from an agricultural watershed indicated that primary runoff can contain viral densities approximating those of receiving waters. Furthermore, viruses attached to suspended colloids made up a large fraction of the total load, particularly in early stages of the storm. At a second field site (stormwater retention pond), RAPD-PCR profiling showed that the viral community of the pond changed dramatically over the course of two intense storms while relatively little change was observed over similar time scales in the absence of disturbance. Comparisons of planktonic and particle-associated viral communities revealed two completely distinct communities, suggesting that particle-associated viruses represent a potentially large and overlooked portion of aquatic viral abundance and diversity. Our findings show that stormwater runoff can quickly change the composition of freshwater microbial communities. Based on these findings, increased storms in the coastal mid-Atlantic region predicted by most climate change models will likely have important impacts on the structure and function of local freshwater microbial communities.


Reduction of fibromyalgia symptoms through intravenous nutrient therapy: results of a pilot clinical trial.
To evaluate the effectiveness of a modified Myers' formula of intravenous nutrient therapy (IVNT) on the symptoms of fibromyalgia (FM) in therapy-resistant FM patients. In this pilot clinical trial, 7 participants with therapy resistant FM were given IVNT once per week for 8 weeks. Patient's pain levels, fatigue, and activities of daily living were evaluated weekly. All participants reported decreased pain levels, decreased fatigue, and increased activities of daily living. Participants noted increased energy levels within 24-48 hours of the initial infusion. At the end of the study, all participants reported increased energy and activities of daily living as well as a 60% reduction in pain (P=.005) and an 80% decrease in fatigue (P=-.005). No participants, however, reported complete or lasting resolution of pain or fatigue. No side effects were reported. Anecdotal reports have indicated benefit for IVNT for patients with chronic pain, including FM. However, except for 2 reports, the medical literature is devoid of any studies of IVNT for the treatment of FM. In this pilot study, 7 participants received IVNT once a week for 8 weeks. All participants had long-standing FM (at least 8 years) and had tried conventional therapies, such as antidepressants, nonsteroidal anti-inflammatory drugs, and exercise, without significant or lasting relief. All had improvement in symptoms and increases in their activities of daily living, although no participant reported complete resolution of symptoms. IVNT appears to be safe to reduce FM symptoms.


Development of the discrimination of spatial phase in infancy.
The ability to discriminate grating patterns, containing the same spatial frequency components but in different phase relationships, has been studied in infants by comparing looking times following habituation to one pattern. The performance of 1-month-olds was compared with that of 2/3-month-old infants. Both age groups could discriminate a set of components in square-wave-phase (fundamental 0.18 c/deg) from components of the same amplitude combined in random phase. However, these compounds differ in peak-to-trough contrast, which infants of both ages could discriminate even for a constant waveform. When contrast was randomized from presentation to presentation, the older group still demonstrated discrimination, implying that they were sensitive to the pattern differences, but the younger group did not. The younger group also failed to demonstrate discrimination between the two waveforms when they were of fixed, matched, peak-to-trough contrast, indicating that the previous absence of discrimination was not simply due to distraction by the contrast variations. We conclude that 1-month-olds are insensitive to the configuration of these compound grating patterns even when they are capable of detecting their components. This loss of spatial information has some analogies with adult peripheral and amblyopic vision. Like other aspects of vision, it shows striking development between 1 and 3 months of age.


Stem cells in the adult pancreas and liver.
Stem cells are undifferentiated cells that can self-renew and generate specialized (functional) cell types. The remarkable ability of stem cells to differentiate towards functional cells makes them suitable modalities in cellular therapy (which means treating diseases with the body's own cells). Potential targets for cellular therapy include diabetes and liver failure. However, in order for stem cells to be clinically useful, we must learn to identify them and to regulate their differentiation. We will use the intestine as a classical example of a stem cell compartment, and then examine the evidence for the existence of adult stem cells in two endodermally derived organs: pancreas and liver. We will review the characteristics of the putative stem cells in these tissues and the transcription factors controlling their differentiation towards functional cell types.


Vitamin D supplementation and upper respiratory tract infections in adolescent swimmers: a randomized controlled trial.
Observational studies identified associations between vitamin D insufficiency (serum 25(OH)D < 30ng·ml-1) and risk of upper respiratory infection (URI). Swimmers are highly prone to URIs, which might hinder their performance. The aim of this study was to examine if vitamin D3 supplementation reduces URI burden in vitamin D-insufficient swimmers. Fifty-five competitive adolescent swimmers with vitamin D insufficiency were randomized to receive vitamin D3 (2,000IU·d-1) or placebo for 12 winter weeks. A URI symptom questionnaire was completed weekly. Serum 25(OH)D concentrations were measured by radio-immunoassay before and after supplementation. We used linear regression to examine the relation between the change in 25(OH)D concentrations during the trial, and the duration and severity of URIs. There were no between-group differences in the frequency, severity, or duration of URIs. Exploratory analyses revealed that in the placebo group only, the change in 25(OH)D concentrations during the trial was highly associated with the duration of URIs (r = -0.90,p < .001), and moderately associated with the severity of URIs (r = -0.65,p = .043). The between-group differences for duration were highly significant. Vitamin D3 supplementation in adolescent swimmers with vitamin D insufficiency did not reduce URI burden. However, larger decreases in serum 25(OH)D concentrations were associated with significantly longer and more severe URI episodes.


Calcium-Binding Proteins S100A8 and S100A9: Investigation of Their Immune Regulatory Effect in Myeloid Cells.
High expression levels of the calcium-binding proteins S100A8 and S100A9 in myeloid cells in kidney transplant rejections are associated with a favorable outcome. Here we investigated the myeloid cell subset expressing these molecules, and their function in inflammatory reactions. Different monocyte subsets were sorted from buffy coats of healthy donors and investigated for S100A8 and S100A9 expression. To characterize S100A9high and S100A9low subsets within the CD14+ classical monocyte subset, intracellular S100A9 staining was combined with flow cytometry (FACS) and qPCR profiling. Furthermore, S100A8 and S100A9 were overexpressed by transfection in primary monocyte-derived macrophages and the THP-1 macrophage cell line to investigate the functional relevance. Expression of S100A8 and S100A9 was primarily found in classical monocytes and to a much lower extent in intermediate and non-classical monocytes. All S100A9+ cells expressed human leukocyte antigen—antigen D related (HLA-DR) on their surface. A small population (<3%) of CD14+ CD11b+ CD33+ HLA-DR− cells, characterized as myeloid derived suppressor cells (MDSCs), also expressed S100A9 to high extent. Overexpression of S100A8 and S00A9 in macrophages led to enhanced extracellular reactive oxygen species (ROS) production, as well as elevated mRNA expression of anti-inflammatory IL-10. The results suggest that the calcium-binding proteins S100A8 and S100A9 in myeloid cells have an immune regulatory effect.


Critical Care Nurses' Reasons for Working or Not Working Overtime.
Around the world, registered nurses are working increasing amounts of overtime. This is particularly true in critical care environments, which experience unpredictable fluctuations in patient volume and acuity, combined with a need for more specialized nurses. To explore critical care nurses' reasons for working or not working overtime. A semistructured interview guide was used to interview 28 frontline nurses from 11 critical care units in Ontario, Canada. Analysis was guided by Thorne's interpretive description methodology. Participants' reasons for working overtime included (1) financial gain (96% of participants); (2) helping and being with colleagues (68%); (3) continuity for nurses and patients (39%); and (4) accelerated career development (39%). Their reasons for not working overtime were (1) feeling tired and tired of being at work (50%); (2) having established plans (71%); and (3) not receiving enough notice (61%). Findings from this study provide important variations and extension of existing literature on the topic, and appear to be the first reported in Canadian critical care units. Additional research is required to understand administrative decision-making processes that lead to the use of overtime.


Rat thoracic duct lymphocytes: types that participate in inflammation.
Newly formed small lymphocytes with a short life-span in the blood are the only cells from thoracic duct lymph which accumulate in acutely inflamed tissue. This conclusion is drawn from studies in which rats with induced peritonieal exudates were injected intravenously with radioactively labeled thoracic duct cells. Radioactivity, originally vested in newly formed donor small lymphocytes, was found later in a small number of similar exudate cells. Small lymphocytes generated 10 days or more before the thoracic ducts were cannulated failed to localize in peritoneal exudates, although the cells moved in large numbers from the blood in to lymph nodes.


Generation of rac3 null mutant mice: role of Rac3 in Bcr/Abl-caused lymphoblastic leukemia.
Numerous studies indirectly implicate Rac GTPases in cancer. To investigate if Rac3 contributes to normal or malignant cell function, we generated rac3 null mutants through gene targeting. These mice were viable, fertile, and lacked an obvious external phenotype. This shows Rac3 function is dispensable for embryonic development. Bcr/Abl is a deregulated tyrosine kinase that causes chronic myelogenous leukemia and Ph-positive acute lymphoblastic leukemia in humans. Vav1, a hematopoiesis-specific exchange factor for Rac, was constitutively tyrosine phosphorylated in primary lymphomas from Bcr/Abl P190 transgenic mice, suggesting inappropriate Rac activation. rac3 is expressed in these malignant hematopoietic cells. Using lysates from BCR/ABL transgenic mice that express or lack rac3, we detected the presence of activated Rac3 but not Rac1 or Rac2 in the malignant precursor B-lineage lymphoblasts. In addition, in female P190 BCR/ABL transgenic mice, lack of rac3 was associated with a longer average survival. These data are the first to directly show a stimulatory role for Rac in leukemia in vivo. Moreover, our data suggest that interference with Rac3 activity, for example, by using geranyl-geranyltransferase inhibitors, may provide a positive clinical benefit for patients with Ph-positive acute lymphoblastic leukemia.


Bicyclic derivatives of hexahydroaporphine: novel agents that lower intraocular pressure.
We have synthesized a number of novel bicyclic hexahydroaporphines containing a phenethylamine moiety for preliminary study as intraocular pressure lowering agents. The target molecules were synthesized in a twelve step process. The final products were secondary or tertiary amines containing either an aromatic methoxy or phenolic substituent. These molecules, in hydrochloride salt form, were assayed in doses ranging from 0.1-1.5%. All products and vehicle were administered topically to one eye of normotensive rabbits and intraocular pressure was measured in both eyes for up to six hours. Four of the five compounds examined produced a significant and, in some cases, prolonged, ocular hypotensive response. Secondary and N-methylated tertiary amines were equally effective, as were compounds containing either the 10-methoxy group of free phenol. Studies are currently in progress to optimize potency and identify functional and molecular mechanisms of action.


Toposome: Source of antimicrobial molecules in the gonads of the sea urchin Lytechinus variegatus (Lamarck, 1816).
Sea urchins live in a challenging environment that requires rapid and efficient responses against pathogens and invaders. This response may be also important in reproductive processes once males and females release their gametes into water. In addition, the gonads are organs with dual function: reproductive organ and nutrient reserve, therefore it needs efficient protective mechanisms to preserve the nutrients as well as the reproductive cells. The aim of this study was to evaluate the presence and characterize antimicrobial molecules in the male and female gonads of the sea urchin Lytechinus variegatus. Through HPLC purification, antimicrobial activity test and mass spectrometry several antimicrobial molecules were found in the gonads of both gender. Computational in silico analyses showed that they are fragments of a glycoprotein called toposome, also known as major yolk protein (MYP) which is one of the major proteins found in the gonads. Although different functions have been reported for this protein, this is the first description of a direct antimicrobial activity in Lytechinus variegatus. The results indicate that when undergoing proteolysis the toposome generates different fragments with antimicrobial activity which may indicate the importance of a rapid defense response strategy against invading microorganisms in the gonads used by both males and females sea urchins.


Stable gene transfer and expression in human primary T cells by the Sleeping Beauty transposon system.
The Sleeping Beauty (SB) transposon system is a nonviral DNA delivery system in which a transposase directs integration of an SB transposon into TA-dinucleotide sites in the genome. To determine whether the SB transposon system can mediate stable gene expression in human T cells, primary peripheral blood lymphocytes (PBLs) were nucleofected with SB vectors carrying a DsRed reporter gene. Plasmids containing the SB transposase on the same molecule as (cis) or on a molecule separate from (trans) the SB transposon mediated long-term and stable reporter gene expression in human primary T cells. Sequencing of transposon:chromosome junctions confirmed that stable gene expression was due to SB-mediated transposition. In other studies, PBLs were successfully transfected using the SB transposon system and shown to stably express a fusion protein consisting of (1) a surface receptor useful for positive T-cell selection and (2) a "suicide" gene useful for elimination of transfected T cells after chemotherapy. This study is the first report demonstrating that the SB transposon system can mediate stable gene transfer in human primary PBLs, which may be advantageous for T-cell-based gene therapies.


Infant massage compared with crib vibrator in the treatment of colicky infants.
To evaluate the effectiveness of infant massage compared with that of a crib vibrator in the treatment of infantile colic. Infants <7 weeks of age and perceived as colicky by their parents were randomly assigned to an infant massage group (n = 28) or a crib vibrator group (n = 30). Three daily intervention periods were recommended in both groups. Parents recorded infant crying and given interventions in a structured cry diary that was kept for 1 week before (baseline) and for 3 weeks during the intervention. Parents were interviewed after the first and third weeks of intervention to obtain their evaluation of the effectiveness of the given massage or crib vibration. At baseline, the mean amount of total crying was 3.6 (standard deviation: 1.4) hours/day in the massage group infants and 4.2 (2.0) hours/day in the vibrator group infants. The mean amount of colicky crying was 2.1 (standard deviation: 1.1) hours/day and 2.9 (1.5) hours/day, respectively. The mean number of daily intervention periods was 2.2 in both groups. Over the 4-week study, the amount of total and colicky crying decreased significantly in both intervention groups. The reduction in crying was similar in the study groups: total crying decreased by a mean 48% in the massage group and by 47% in the vibrator group, and colicky crying decreased by 64% and 52%, respectively. The amount of other crying (total crying minus colicky crying) remained stable in both groups over the intervention. Ninety-three percent of the parents in both groups reported that colic symptoms decreased over the 3-week intervention, and 61% of the parents in the massage group and 63% of the parents in the crib vibrator group perceived the 3-week intervention as colic reducing. Infant massage was comparable to the use of a crib vibrator in reducing crying in colicky infants. We suggest that the decrease of total and colicky crying in the present study reflects more the natural course of early infant crying and colic than a specific effect of the interventions.


Combination of bevacizumab and acyclic retinoid inhibits the growth of hepatocellular carcinoma xenografts.
The prognosis of patients with hepatocellular carcinoma (HCC) is poor and the development of effective treatments for this malignancy, including combination chemotherapy, is required. This study examined the possible combined inhibitory effects of bevacizumab, an anti-vascular endothelial growth factor monoclonal antibody, and acyclic retinoid (ACR), which can prevent the development of HCC, on the growth of Huh7 human HCC cells. Xenograft tumors were produced by subcutaneously injecting Huh7 cells into nude mice. Starting 1 wk after the tumor cell injection, the mice were treated with bevacizumab alone (5 mg/kg body weight, subcutaneous injection, twice a week), ACR alone (given in a diet containing 0.03%), or their combination for 6 wk, and the effects of these regimens on xenograft growth were examined. Combined treatment with bevacizumab plus ACR significantly suppressed the growth of Huh7 xenografts. The combination of these agents significantly inhibited the phosphorylation of the Akt protein in tumor tissues. With combination therapy, the population of Ki-67-positive cells in xenografts decreased, while that of TUNEL-positive cells increased. The combination of bevacizumab and ACR exerts growth-suppressing effects on HCC cells by inhibiting cell proliferation and inducing apoptosis. This combination might be an effective regimen for the treatment of HCC.


Randomized Multicenter Placebo-Controlled Trial of Omega-3 Fatty Acids for the Control of Aromatase Inhibitor-Induced Musculoskeletal Pain: SWOG S0927.
Musculoskeletal symptoms are the most common adverse effects of aromatase inhibitors (AIs) and can result in decreased quality of life and discontinuation of therapy. Omega-3 fatty acids (O3-FAs) can be effective in decreasing arthralgia resulting from rheumatologic conditions and reducing serum triglycerides. Women with early-stage breast cancer receiving an AI who had a worst joint pain/stiffness score ≥ 5 of 10 using the Brief Pain Inventory-Short Form (BPI-SF) were randomly assigned to receive either O3-FAs 3.3 g or placebo (soybean/corn oil) daily for 24 weeks. Clinically significant change was defined as ≥ 2-point drop from baseline. Patients also completed quality-of-life (Functional Assessment of Cancer Therapy-Endocrine Symptoms) and additional pain/stiffness assessments at baseline and weeks 6, 12, and 24. Serial fasting blood was collected for lipid analysis. Among 262 patients registered, 249 were evaluable, with 122 women in the O3-FA arm and 127 in the placebo arm. Compared with baseline, the mean observed BPI-SF score decreased by 1.74 points at 12 weeks and 2.22 points at 24 weeks with O3-FAs and by 1.49 and 1.81 points, respectively, with placebo. In a linear regression adjusting for the baseline score, osteoarthritis, and taxane use, adjusted 12-week BPI-SF scores did not differ by arm (P = .58). Triglyceride levels decreased in patients receiving O3-FA treatment and remained the same for those receiving placebo (P = .01). No between-group differences were seen for HDL, LDL, or C-reactive protein. We found a substantial (> 50%) and sustained improvement in AI arthralgia for both O3-FAs and placebo but found no meaningful difference between the groups.


Erythrocyte fatty acids and breast cancer risk: a case-control study in Shanghai, China.
The role of individual fatty acids in the development and progression of breast cancer is unclear. Although in vitro and animal experiments have supported an inverse association between intake of long chain n-3 fatty acids [primarily eicosapentaenoic acid (EPA) and docosahexaenoic acid] and breast cancer risk, findings from population studies are inconsistent. Recent studies have also shown associations between the ratio of saturated to monounsaturated fatty acids (SI) and breast cancer risk. The SI reflects the activity of several genes involved in lipid metabolism, including fatty acid synthase and steroyl coenzyme-A desaturase, that have been shown to be overexpressed in breast cancer. The purpose of this analysis was to determine the association between erythrocyte fatty acid concentrations and breast cancer risk among women participating in a randomized trial of breast self-examination in Shanghai, China. We conducted a case-control study. Erythrocyte fatty acid concentrations were determined in specimens from 322 women with histologically confirmed breast cancer and 1030 frequency age-matched control women. We report a significant direct association among palmitic, gamma-linolenic, palmitoleic, and vaccenic acids and risk of breast cancer. Total n-3 fatty acids, EPA, and the SI for palmitic to palmitoleic acid were associated with significantly lower risk of breast cancer. Our results support a protective effect of n-3 fatty acids on breast cancer risk and provide additional evidence for the importance of evaluating the ratio of fatty acids when evaluating diet and breast cancer risk.


Rheumatological manifestations in patients hospitalized with COVID-19 in a tertiary care setting.
Corona virus disease 2019 (COVID-19) is a pandemic affecting over 150 million people worldwide since its emergence in Wuhan, China in December 2019, and is leading to over 600 thousand deaths. Severe acute respiratory syndrome novel coronavirus 2 (SARS-nCoV-2) causes a wide range of symptoms by affecting different organ systems of the body. This study was designed to specifically look for its rheumatological manifestations. It was a cross-sectional study conducted in the corona complex MTI Lady Reading Hospital Peshawar. A total of 141 COVID-19 positive hospitalized patients requiring oxygen therapy and having no previous rheumatological diseases were enrolled and studied for any rheumatological manifestations. There were 88 (62.4%) males and 53 (37.6%) females, with age range from 27-88 years, and mean age of 58.5±11.5 SD. Arthralgia and myalgia were found very common, occurring in 128 (90.8%) and 119 (84.4%) of patients. Arthritis was not found in any patient. Arthralgia and myalgia were reported frequently in patients with severe COVID-19 requiring oxygen therapy, while arthritis was not present even in a single patient as is reported in other viral infections.


Genetic Variants and Systemic Complement Activation Levels Are Associated With Serum Lipoprotein Levels in Age-Related Macular Degeneration.
Genetic variants in genes encoding components of lipid metabolism have been associated with AMD. The aims of this study were to evaluate the relation of these genetic variants with serum lipid levels in AMD in a large case-control cohort (n = 3070) and to test for correlations between lipids and complement activation. Single nucleotide polymorphisms (SNPs) in eight lipid metabolism genes, previously described to be associated with AMD, were genotyped and tested for their association in our case-control cohort. Serum apolipoprotein B (ApoB), apolipoprotein AI (Apo-AI), cholesterol, triglycerides (TG), high-density lipoprotein-cholesterol (HDLC), and complement activation levels (C3d/C3) were measured and tested for association with AMD. Non-HDL cholesterol and LDL were inferred based on the measurements of the other lipids and lipoproteins. General linear models and χ2 tests were used to evaluate the relation of SNPs and lipids/lipoproteins to the disease as well as their interrelations. Significant genotypic associations with AMD were observed for SNPs in CETP, APOE, and FADS1. The serum levels of Apo-AI and HDLC were significantly higher in patients compared with controls. Triglycerides (TG) levels were lower in AMD compared with controls. A cumulative effect was observed for APOE and CETP genotypes on HDLC and Apo-AI levels. Complement activation levels correlated positively with HDLC and Apo-AI, and negatively with TG. Both the lipids/lipoproteins and the complement activation levels associate independently to AMD. This study bridges the gap between genetic associations and physiological lipid levels in AMD. Additionally, the observed correlations between complement activation and lipid levels link two major systems that previously were always assessed independently.


Spectral-profile-based algorithm for hemoglobin oxygen saturation determination from diffuse reflectance spectra.
Variations of hemoglobin (Hb) oxygenation in tissue provide important indications concerning the physiological conditions of tissue, and the data related to these variations are of intense interest in medical research as well as in clinical care. In this study, we derived a new algorithm to estimate Hb oxygenation from diffuse reflectance spectra. The algorithm was developed based on the unique spectral profile differences between the extinction coefficient spectra of oxy-Hb and deoxy-Hb within the visible wavelength region. Using differential wavelet transformation, these differences were quantified using the locations of certain spectral features, and, then, they were related to the oxygenation saturation level of Hb. The applicability of the algorithm was evaluated using a set of diffuse reflectance spectra produced by a Monte Carlo simulation model of photon migration and by tissue phantoms experimentally. The algorithm was further applied to the diffuse reflectance spectra acquired from in vivo experiments to demonstrate its clinical utility. The validation and evaluation results concluded that the algorithm is applicable to various tissue types (i.e., scattering properties) and can be easily used in conjunction with a diverse range of probe geometries for real-time monitoring of Hb oxygenation.


An international study of clinical education practices in speech-language pathology.
This study identified the clinical placement and supervisory models used within 45 speech-language pathology professional preparation programs in seven English-speaking countries and the drivers influencing the adoption of these models. University personnel completed an on-line survey about the placement and supervisory models used in their programs, their opinions regarding the effectiveness of the models in developing student competency, and the factors influencing the adoption of different models. Responses from the survey were analysed through descriptive statistics and thematic analysis. "Traditional" placement models such as block and weekly placements, as well as traditional supervisory models utilizing a speech-language pathologist as supervisor in a 1:1 student-to-supervisor ratio were the most widely used in the clinical education of speech-language pathologists. "Non-traditional" models, where the overall structure of the practicum experience or the delivery of supervision within the practicum experience has been altered from the traditional models, were used with differing frequency across countries. The major influences on the adoption of non-traditional models included attitudes about the effectiveness of the models, availability of placements, standards of professional associations, and student learning outcomes, particularly in relation to exposure to clinical sub-groups. Further evidence regarding the effectiveness of both traditional and non-traditional placement and supervisory models needs to be developed. To develop this evidence, consistent terminology and a shared understanding of terminology needs to be established.


Ecoimmunology and microbial ecology: Contributions to avian behavior, physiology, and life history.
Bacteria have had a fundamental impact on vertebrate evolution not only by affecting the evolution of the immune system, but also generating complex interactions with behavior and physiology. Advances in molecular techniques have started to reveal the intricate ways in which bacteria and vertebrates have coevolved. Here, we focus on birds as an example system for understanding the fundamental impact bacteria have had on the evolution of avian immune defenses, behavior, physiology, reproduction and life histories. The avian egg has multiple characteristics that have evolved to enable effective defense against pathogenic attack. Microbial risk of pathogenic infection is hypothesized to vary with life stage, with early life risk being maximal at either hatching or fledging. For adult birds, microbial infection risk is also proposed to vary with habitat and life stage, with molt inducing a period of increased vulnerability. Bacteria not only play an important role in shaping the immune system as well as trade-offs with other physiological systems, but also for determining digestive efficiency and nutrient uptake. The relevance of avian microbiomes for avian ecology, physiology and behavior is highly topical and will likely impact on our understanding of avian welfare, conservation, captive breeding as well as for our understanding of the nature of host-microbe coevolution.


Honokiol in combination with radiation targets notch signaling to inhibit colon cancer stem cells.
Cancer stem cells are implicated in resistance to ionizing radiation (IR) and chemotherapy. Honokiol, a biphenolic compound has been used in traditional Chinese medicine for treating various ailments. In this study, we determined the ability of honokiol to enhance the sensitivity of colon cancer stem cells to IR. The combination of honokiol and IR suppressed proliferation and colony formation while inducing apoptosis of colon cancer cells in culture. There were also reduced numbers and size of spheroids, which was coupled with reduced expression of cancer stem cell marker protein DCLK1. Flow cytometry studies confirmed that the honokiol-IR combination reduced the number of DCLK1+ cells. In addition, there were reduced levels of activated Notch-1, its ligand Jagged-1, and the downstream target gene Hes-1. Furthermore, expression of components of the Notch-1 activating γ-secretase complex, presenilin 1, nicastrin, Pen2, and APH-1 was also suppressed. On the other hand, the honokiol effects were mitigated when the Notch intracellular domain was expressed. To determine the effect of honokiol-IR combination on tumor growth in vivo, nude mice tumor xenografts were administered honokiol intraperitoneally and exposed to IR. The honokiol-IR combination significantly inhibited tumor xenograft growth. In addition, there were reduced levels of DCLK1 and the Notch signaling-related proteins in the xenograft tissues. Together, these data suggest that honokiol is a potent inhibitor of colon cancer growth that targets the stem cells by inhibiting the γ-secretase complex and the Notch signaling pathway. These studies warrant further clinical evaluation for the combination of honokiol and IR for treating colon cancers.


Feeling happy when feeling down: The effectiveness of positive mental imagery in dysphoria.
Mental imagery can evoke strong emotional responses, but imagery perspective can influence the response, with observer perspective reducing emotionality. This is important provided that positive imagery can be an effective mood repair strategy in healthy individuals. However, (sub-clinical) depressed individuals tend to spontaneously adopt an observer perspective. We investigated whether positive imagery would result in a similar emotional response in dysphoric and non-dysphoric individuals when instructed and trained to use field perspective imagery. Additionally, we compared the emotional response in dysphoric individuals who received instructions to dysphoric individuals who received no instructions on processing mode during positive memory recall. Dysphoric and non-dysphoric individuals completed a mood induction procedure imagining positive or neutral memories. They received instructions and practice in the use of field perspective imagery. An additional control group of dysphoric individuals recalled positive memories without receiving instructions on processing mode. Dysphoric and non-dysphoric individuals who received instructions on field perspective imagery reported similar use of field and observer perspective imagery, and a similar positive emotional response. Dysphoric individuals who did not receive specific instructions, as compared to those who did, reported greater use of observer perspective and lower levels of positive affect afterwards. A dysphoric sample limits generalization to clinically depressed individuals, although these individuals are at risk for developing depression. However, mental imagery used in relapse prevention is likely targeting sub-clinical populations. Providing practice in field perspective imagery could potentially improve the effectiveness of positive memory recall as a mood repair strategy in (sub-clinically) depressed individuals, and may therefore have important therapeutic benefits.


A quasi-dense approach to surface reconstruction from uncalibrated images.
This paper proposes a quasi-dense approach to 3D surface model acquisition from uncalibrated images. First, correspondence information and geometry are computed based on new quasi-dense point features that are resampled subpixel points from a disparity map. The quasi-dense approach gives more robust and accurate geometry estimations than the standard sparse approach. The robustness is measured as the success rate of full automatic geometry estimation with all involved parameters fixed. The accuracy is measured by a fast gauge-free uncertainty estimation algorithm. The quasi-dense approach also works for more largely separated images than the sparse approach, therefore, it requires fewer images for modeling. More importantly, the quasidense approach delivers a high density of reconstructed 3D points on which a surface representation can be reconstructed. This fills the gap of insufficiency of the sparse approach for surface reconstruction, essential for modeling and visualization applications. Second, surface reconstruction methods from the given quasi-dense geometry are also developed. The algorithm optimizes new unified functionals integrating both 3D quasi-dense points and 2D image information, including silhouettes. Combining both 3D data and 2D images is more robust than the existing methods using only 2D information or only 3D data. An efficient bounded regularization method is proposed to implement the surface evolution by level-set methods. Its properties are discussed and proven for some cases. As a whole, a complete automatic and practical system of 3D modeling from raw images captured by hand-held cameras to surface representation is proposed. Extensive experiments demonstrate the superior performance of the quasi-dense approach with respect to the standard sparse approach in robustness, accuracy, and applicability.


Independent colonization of multiple urban centres by a formerly forest specialist bird species.
Urban areas are expanding rapidly, but a few native species have successfully colonized them. The processes underlying such colonization events are poorly understood. Using the blackbird Turdus merula, a former forest specialist that is now one of the most common urban birds in its range, we provide the first assessment of two contrasting urban colonization models. First, that urbanization occurred independently. Second, that following initial urbanization, urban-adapted individuals colonized other urban areas in a leapfrog manner. Previous analyses of spatial patterns in the timing of blackbird urbanization, and experimental introductions of urban and rural blackbirds to uncolonized cities, suggest that the leapfrog model is likely to apply. We found that, across the western Palaearctic, urban blackbird populations contain less genetic diversity than rural ones, urban populations are more strongly differentiated from each other than from rural populations and assignment tests support a rural source population for most urban individuals. In combination, these results provide much stronger support for the independent urbanization model than the leapfrog one. If the former model predominates, colonization of multiple urban centres will be particularly difficult when urbanization requires genetic adaptations, having implications for urban species diversity.


RNA polymerase B subunit gene mutations in biofilm-embedded methicillin-resistant Staphylococcus aureus following rifampin treatment.
This study was conducted to compare the mutation rates of different rpoB sites and rifampin minimum inhibitory concentration (MIC) changes prior to and after rifampin therapy for biofilm-embedded methicillin-resistant Staphylococcus aureus (MRSA) isolates. The screening of rifampin-resistant MRSA isolates, from the biofilm at Day 5 with or without exposure to the susceptible breakpoint concentration of rifampin recommended by the Clinical and Laboratory Standards Institute (1 mg/L), was conducted using agar plates containing rifampin. A partial fragment of RNA polymerase B subunit gene (rpoB), including clusters I and II, was amplified and sequenced. The rifampin MIC values and mutation frequencies at different sites of rpoB were measured and evaluated in rifampicin-resistant isolates. Rifampin-resistant mutants could be selected from all of 39 randomly selected rifampin-susceptible MRSA isolates in the biofilm model. The spontaneous mutation frequency ranged from 1.00 × 10(-4) to 3.85 × 10(-7). Mutation at codon 481 was most commonly found at 35 (89.7%) of 39 MRSA isolates. Without rifampin induction, the MIC ranged between 0.125 mg/L and1024 mg/L and mutation sites included cluster I 464, 466, 468, 471, 474, 477, 481, 484, 486 and cluster II 519, 527, 529 with the percentage of 471 (35.9%), 477 (33.3%), 481 (53.8%), and 484 (35.9%). Conversely, with the induction of rifampin, the MIC value ranged ∼256-1024 mg/L. The mutation sites that were more concentrated included 468 (17.9%), 477 (30.8%), 481 (89.7%), 484 (17.9%), and 486 (33.3%). We documented high rifampin resistance induction activity when MRSA was engaged in biofilm with rifampin exposure. Monotherapy seems to be inadequate for MRSA in biofilm. There is an urgent need for developing effective combination therapies with less rifampin resistance-inducing activities for treating MRSA in biofilms.


Real-Time Decoding of Attentional States Using Closed-Loop EEG Neurofeedback.
Sustained attention is a cognitive ability to maintain task focus over extended periods of time (Mackworth, 1948; Chun, Golomb, & Turk-Browne, 2011). In this study, scalp electroencephalography (EEG) signals were processed in real time using a 32 dry-electrode system during a sustained visual attention task. An attention training paradigm was implemented, as designed in DeBettencourt, Cohen, Lee, Norman, and Turk-Browne (2015) in which the composition of a sequence of blended images is updated based on the participant's decoded attentional level to a primed image category. It was hypothesized that a single neurofeedback training session would improve sustained attention abilities. Twenty-two participants were trained on a single neurofeedback session with behavioral pretraining and posttraining sessions within three consecutive days. Half of the participants functioned as controls in a double-blinded design and received sham neurofeedback. During the neurofeedback session, attentional states to primed categories were decoded in real time and used to provide a continuous feedback signal customized to each participant in a closed-loop approach. We report a mean classifier decoding error rate of 34.3% (chance = 50%). Within the neurofeedback group, there was a greater level of task-relevant attentional information decoded in the participant's brain before making a correct behavioral response than before an incorrect response. This effect was not visible in the control group (interaction p=7.23e-4), which strongly indicates that we were able to achieve a meaningful measure of subjective attentional state in real time and control participants' behavior during the neurofeedback session. We do not provide conclusive evidence whether the single neurofeedback session per se provided lasting effects in sustained attention abilities. We developed a portable EEG neurofeedback system capable of decoding attentional states and predicting behavioral choices in the attention task at hand. The neurofeedback code framework is Python based and open source, and it allows users to actively engage in the development of neurofeedback tools for scientific and translational use.


Dupilumab in Children with Uncontrolled Moderate-to-Severe Asthma.
Children with moderate-to-severe asthma continue to have disease complications despite the receipt of standard-of-care therapy. The monoclonal antibody dupilumab has been approved for the treatment of adults and adolescents with asthma as well as with other type 2 inflammatory diseases. In this 52-week phase 3, randomized, double-blind, placebo-controlled trial, we assigned 408 children between the ages of 6 and 11 years who had uncontrolled moderate-to-severe asthma to receive a subcutaneous injection of dupilumab (at a dose of 100 mg for those weighing ≤30 kg and 200 mg for those weighing >30 kg) or matched placebo every 2 weeks. All the children continued to receive a stable dose of standard background therapy. The primary end point was the annualized rate of severe asthma exacerbations. Secondary end points included the change from baseline in the percentage of predicted prebronchodilator forced expiratory volume in 1 second (ppFEV1) at week 12 and in the score on the Asthma Control Questionnaire 7 Interviewer-Administered (ACQ-7-IA) at week 24. End points were evaluated in the two primary efficacy populations who had either a type 2 inflammatory asthma phenotype (≥150 blood eosinophils per cubic millimeter or a fraction of exhaled nitric oxide of ≥20 ppb at baseline) or a blood eosinophil count of at least 300 cells per cubic millimeter at baseline. In patients with the type 2 inflammatory phenotype, the annualized rate of severe asthma exacerbations was 0.31 (95% confidence interval [CI], 0.22 to 0.42) with dupilumab and 0.75 (95% CI, 0.54 to 1.03) with placebo (relative risk reduction in the dupilumab group, 59.3%; 95% CI, 39.5 to 72.6; P<0.001). The mean (±SE) change from baseline in the ppFEV1 was 10.5±1.0 percentage points with dupilumab and 5.3±1.4 percentage points with placebo (mean difference, 5.2 percentage points; 95% CI, 2.1 to 8.3; P<0.001). Dupilumab also resulted in significantly better asthma control than placebo (P<0.001). Similar results were observed in the patients with an eosinophil count of at least 300 cells per cubic millimeter at baseline. The incidence of serious adverse events was similar in the two groups. Among children with uncontrolled moderate-to-severe asthma, those who received add-on dupilumab had fewer asthma exacerbations and better lung function and asthma control than those who received placebo. (Funded by Sanofi and Regeneron Pharmaceuticals; Liberty Asthma VOYAGE ClinicalTrials.gov number, NCT02948959.).


Quantum noise in fresnel zone plate imaging.
A Fresnel zone plate, used as a coded aperture, offers a great advantage in geometric collection efficiency over a conventional pinhole or collimator. We present a detailed analysis of the signal-to-noise ratio (SNR) of a quantum-limited zone plate camera. The magnitude and spatial distribution of the noise field and its dependence on the source distribution and the characteristics of the optical processing system are derived. It is shown that the largest SNR advantage occurs for a point source, while for very large, uniform sources there may be a slight net disadvantage to using a zone plate. It is also shown that optical processing does not give the highest possible SNR.


Hospitalizations among children involved in the child protection system: A long-term birth cohort study from infancy to adulthood using administrative data.
Despite considerable health consequences of child abuse and neglect, there is limited evidence on hospitalizations in this population. To describe frequency and reasons for hospitalization by lifetime child protection system (CPS) involvement. 608,540 children born from January 1, 1986 to June 30, 2017 in South Australia, Australia METHODS: Using linked administrative data on CPS involvement and hospitalizations, we descriptively examined cumulative incidence, cumulative count and reasons for hospitalization from infancy to early adulthood by CPS involvement. By 16.5 years of age, cumulative incidence of ever-being hospitalized was 58% (95% CI 58-58) for children with no CPS involvement and significantly higher (72% (95% CI 71-73) to 88% (95% CI 86-90), P < 0.001) among those with different levels of CPS involvement. Cumulative mean counts of hospitalizations were highest at every age for those placed in out-of-home-care (reaching 7.7 by 16.5 years), almost four times higher than for children with no CPS involvement (2.0 by 16.5 years). Most frequent reasons for hospitalizations were similar across CPS involvement in the early years. From adolescence through early adulthood, mental health, and 'injury, poisoning or toxic effects of drugs' were frequent reasons for hospitalization among individuals with CPS involvement. This study highlights the vulnerability of children who have been maltreated, or are at risk of maltreatment, and the urgency of implementing effective preventive strategies early in life including consideration of adequate responses of child protection services. Frequent hospitalizations for mental health and injury confirms the potentially preventable nature of these hospitalizations.


Characterization of follistatin-related gene as a negative regulatory factor for activin family members during mouse heart development.
Follistatin-related gene (FLRG) encodes a secretory glycoprotein that has characteristic cysteine-rich follistatin domains. FLRG protein binds to and neutralizes several transforming growth factor-beta (TGF-beta) superfamily members, including myostatin (MSTN), which is a potent negative regulator of skeletal muscle mass. We have previously reported that FLRG was abundantly expressed in fetal and adult mouse heart. In this study, we analyzed the expression of FLRG mRNA during mouse heart development. FLRG mRNA was continuously expressed in the embryonic heart, whereas it was very low in skeletal muscles. By contrast, MSTN mRNA was highly expressed in embryonic skeletal muscles, whereas the expression of MSTN mRNA was rather low in the heart. In situ hybridization and immunohistochemical analysis revealed that FLRG expressed in smooth muscle of the aorta and pulmonary artery, valve leaflets of mitral and tricuspid valves, and cardiac muscles in the ventricle of mouse embryonic heart. However, MSTN was expressed in very limited areas, such as valve leaflets of pulmonary and aortic valves, the top of the ventricular and atrial septa. Interestingly, the expression of MSTN was complementary to that of FLRG, especially in the valvular apparatus. Biochemical analyses with surface plasmon resonance biosensor and reporter assays demonstrated that FLRG hardly dissociates from MSTN and activin once it bound to them, and efficiently inhibits these activities. Our results suggest that FLRG could function as a negative regulator of activin family members including MSTN during heart development.


Gain-of-function mutations in a human calmodulin-like protein identify residues critical for calmodulin action in yeast.
A human epithelial cell-specific transcript (NB-1) encodes a calmodulin-like protein (hCLP), which is identical in length and 85% identical in amino acid sequence to authentic human calmodulin (hCaM). Although hCaM shares only 60% amino acid sequence identity with yeast calmodulin (CMD1 gene product), hCaM was able to substitute functionally for Cmd1 in yeast cells. In contrast, hCLP was unable to support either spore germination or vegetative growth in Cmd1-deficient yeast cells, even when stably expressed at a level at least an order of magnitude above that of hCaM. Thus, hCLP provides an indicator protein for discerning those residues that are critical for calmodulin function in vivo. In addition to 20 conservative amino acid replacements, hCLP differs from hCaM (and other vertebrate calmodulins that are able to complement a cmd1 null mutation) by only three nonconservative substitutions. Site-directed mutagenesis was used to convert these three positions back to residues more typical of those found in authentic calmodulins and to prepare all possible combinations of these three mutations, specifically: three single mutants (R58V, R112N, and A128E), three double mutants (R58V A128E, R112N A128E, and R58V R112N), and the triple mutant (R58V R112N A128E). The triple mutant and one of the double mutants (R58V A128E) were able to restore an apparently normal growth rate to a cmd1 delta strain, indicating that the altered hCLPs have acquired the ability to behave as functional calmodulins in yeast. The other two double mutants were able to support growth of Cmd1-deficient cells only weakly, but cells expressing the R112N A128E mutant grew noticeably better than those expressing the R58V R112N mutant. Remarkably, one single mutant (A128E), but not the other two single mutants, was also reproducibly able to support weak growth of a cmd1 delta strain. The properties of these gain-of-function, or neomorphic, mutations implicate E128, and to a lesser extent V58, as residues critical for calmodulin action in vivo. Molecular modeling of these positions within the structure of a Ca(2+)-calmodulin.peptide complex indicates that E128 projects directly into the central cavity occupied by the bound peptide. Thus, E128 may contribute a contact that is vital for the interaction of Cmd1 with one or more of the targets that are essential for yeast cell growth.


Nonenhanced helical CT and US in the emergency evaluation of patients with renal colic: prospective comparison.
To compare nonenhanced helical computed tomography (CT) with ultrasonography (US) for the depiction of urolithiasis. During 9 months, 45 patients (mean age, 44 years; mean weight, 92.5 kg) prospectively underwent both nonenhanced helical CT (5-mm collimation; pitch of 1.5) and US of the kidneys, ureters, and bladder. US evaluation included a careful search for ureteral calculi. Presence of calculi and obstruction and incidental diagnoses were recorded. Clinical, surgical, and/or imaging follow-up data were obtained in all patients. The McNemar test was used to compare groups. Diagnoses included 23 ureteral calculi and one each of renal cell carcinoma, appendicitis, ureteropelvic junction obstruction, renal subcapsular hematoma, cholelithiasis, medullary calcinosis, and myelolipoma. CT depicted 22 of 23 ureteral calculi (sensitivity, 96%). US depicted 14 of 23 ureteral calculi (sensitivity, 61%). Differences in sensitivity were statistically significant (P: =.02). Specificity for each technique was 100%. When modalities were compared for the detection of any clinically relevant abnormality (eg, unilateral hydronephrosis and/or urolithiasis in patients with an obstructing calculus), sensitivities of US and CT increased to 92% and 100%, respectively. One case of appendicitis was missed at US, whereas medullary calcinosis and myelolipoma were missed at CT. Nonenhanced CT has a higher sensitivity for the detection of ureteral calculi compared with US.


Development of a rapid serological assay for the diagnosis of strongyloidiasis using a novel diffraction-based biosensor technology.
Strongyloidiasis is a persistent human parasitic infection caused by the intestinal nematode, Strongyloides stercoralis. The parasite has a world-wide distribution, particularly in tropical and subtropical regions with poor sanitary conditions. Since individuals with strongyloidiasis are typically asymptomatic, the infection can persist for decades without detection. Problems arise when individuals with unrecognized S. stercoralis infection are immunosuppressed, which can lead to hyper-infection syndrome and disseminated disease with an associated high mortality if untreated. Therefore a rapid, sensitive and easy to use method of diagnosing Strongyloides infection may improve the clinical management of this disease. An immunological assay for diagnosing strongyloidiasis was developed on a novel diffraction-based optical bionsensor technology. The test employs a 31-kDa recombinant antigen called NIE derived from Strongyloides stercoralis L3-stage larvae. Assay performance was tested using retrospectively collected sera from patients with parasitologically confirmed strongyloidiasis and control sera from healthy individuals or those with other parasitoses including schistosomiasis, trichinosis, echinococcosis or amebiasis who were seronegative using the NIE ELISA assay. If we consider the control group as the true negative group, the assay readily differentiated S. stercoralis-infected patients from controls detecting 96.3% of the positive cases, and with no cross reactivity observed in the control group These results were in excellent agreement (κ = 0.98) with results obtained by an NIE-based enzyme-linked immunosorbent assay (ELISA). A further 44 sera from patients with suspected S. stercoralis infection were analyzed and showed 91% agreement with the NIE ELISA. In summary, this test provides high sensitivity detection of serum IgG against the NIE Strongyloides antigen. The assay is easy to perform and provides results in less than 30 minutes, making this platform amenable to rapid near-patient screening with minimal technical expertise.


Impact of CYP2C19 and CYP2C9 gene polymorphisms on sodium valproate plasma concentration in patients with epilepsy.
Valproic acid (VPA) is a broad spectrum anticonvulsant drug, which could be partially metabolised by cytochrome P450 (CYP) 2C9 and 2C19 enzymes. This study was designed to investigate the relationship between CYP2C19 and CYP2C9 gene polymorphisms and the plasma concentrations of VPA in subjects with epilepsy. Eighty-three subjects with epilepsy aged 18-92 years were enrolled in this study. All were treated with sustained-release VPA monotherapy. Based on the genotypes of CYP2C19 and the ability to metabolise substrates, the subjects were divided into poor metabolisers, intermediate metabolisers and extensive metabolisers. Sanger sequencing was used to detect the genotypic and allelic frequencies of CYP2C19 (*1, *2 and *3) and CYP2C9 (*13) of the patients. Automatic immunity analysis was used to find steady-state trough plasma concentrations of VPA. By adjusting the plasma concentrations of VPA with body weight and total daily dose of VPA, the concentration-to-dose ratio of VPA (CDRV) was obtained. Data were analysed using SPSS software. The genetic frequencies of CYP2C19*2, CYP2C19*3 and CYP2C9*13 were 33.1%, 3.0% and 5.4%, respectively, among patients with epilepsy from Yunnan province, China who used VPA therapy. The CDRV was significantly lower in the CYP2C19 extensive metabolisers (3.33±1.78) than it was in the CYP2C19 intermediate metabolisers (4.45±1.42) and the CYP2C19 poor metabolizers (6.64±1.06). The CYP2C19*2 and CYP2C19*3 alleles were correlated with the plasma VPA concentration, while the CYP2C9*13 allele had no effect on the plasma VPA concentration (p=0.809). The genetic polymorphisms of CYP2C19 significantly affect the VPA plasma concentration, and the dosage of VPA for intermediate and poor metabolisers could be lower than for extensive metabolisers. CYP2C9*13 carrier was not closely related to plasma concentrations of VPA in patients with epilepsy.


Dual femtosecond laser multiheterodyne optical coherence tomography.
A time domain optical coherence tomography (OCT) system without moving parts is described, which is based on multiheterodyning utilizing two mode-locked femtosecond lasers. By synchronizing the two lasers to slightly different repetition rates and coupling to an interferometric OCT setup, we obtain amplitude-modulated beat signals representing the structure of the specimen under investigation. Our system is suitable for biological imaging as well as technical applications. We demonstrate high axial imaging depths of 150 mm with up to 5000 axial scans per second, achieving equivalent path scanning velocities of 750 m/s.


PRL2 phosphatase promotes oncogenic KIT signaling in leukemia cells through modulating CBL phosphorylation.
Receptor tyrosine kinase KIT is frequently activated in acute myeloid leukemia (AML). While high PRL2 (PTP4A2) expression is correlated with activation of SCF/KIT signaling in AML, the underlying mechanisms are not fully understood. We discovered that inhibition of PRL2 significantly reduces the burden of oncogenic KIT-driven leukemia and extends leukemic mice survival. PRL2 enhances oncogenic KIT signaling in leukemia cells, promoting their proliferation and survival. We found that PRL2 dephosphorylates CBL at tyrosine 371 and inhibits its activity toward KIT, leading to decreased KIT ubiquitination and enhanced AKT and ERK signaling in leukemia cells. Implications: Our studies uncover a novel mechanism that fine-tunes oncogenic KIT signaling in leukemia cells and will likely identify PRL2 as a novel therapeutic target in AML with KIT mutations.


Virtual Reality (VR)-Based Environmental Enrichment in Older Adults with Mild Cognitive Impairment (MCI) and Mild Dementia.
Despite an alarming rise in the global prevalence of dementia, the available modalities for improving cognition and mental wellbeing of dementia patients remain limited. Environmental enrichment is an experimental paradigm that has shown promising anti-depressive and memory-enhancing effects in pre-clinical studies. However, its clinical utility has remained limited due to the lack of effective implementation strategies. The primary objective of this study was to evaluate the usability (tolerability and interactivity) of a long-term virtual reality (VR)- based environmental enrichment training program in older adults with mild cognitive impairment (MCI) and mild dementia. A secondary objective was to assess the effect of VR-based environmental enrichment on stabilization of cognitive functioning and improvement of mental wellbeing in older adults with MCI and mild dementia. A total of seven participants (four patients with MCI and three with mild dementia) received biweekly VR-based environmental enrichment over a course of 6 months. The tolerability and interactivity of the participants in the VR training was serially assessed via virtual reality sickness questionnaire (VRSQ) and recording of input-error ratio. Cognitive functioning was assessed through Montreal cognitive assessment (MoCA) before and after the study. Mental wellbeing was assessed through Warwick-Edinburgh Mental Well Being Scale (WEMWBS). VR-based environmental enrichment was well-tolerated by the patients with significant decrease in VRSQ scores (p < 0.01) and input-error ratio (p < 0.001) overtime. VR training was also effective in stabilization of MoCA scores over the course of therapy (non-significant difference in the MoCA scores before and after the therapy) and was associated with a trend (p < 0.1) towards improvement in WEMWBS scores between the first and the last assessments. Qualitative observations by the care-givers further corroborated a noticeable improvement in mental wellbeing of patients. This pilot study shows that VR can be a feasible, tolerable, and potentially effective tool in long-term support of older adults with MCI and mild dementia.


The Effects of Low-Dose-Rate γ-irradiation on Forced Swim Test-Induced Immobility and Oxidative Stress in Mice.
The forced swim test (FST) induces immobility in mice. Low-dose (high-dose-rate) X-irradiation inhibits FSTinduced immobility in mice due to its antioxidative function. We evaluated the effects of low-dose γ-irradiation at a low-dose-rate on the FST-induced depletion of antioxidants in mouse organs. Mice received whole-body low-dose-rate (0.6 or 3.0 mGy/h) of low-dose γ-irradiation for 1 week, followed by daily FSTs (5 days). The immobility rate on day 2 compared to day 1 was significantly lower in the 3.0 mGy/h irradiated mice than in sham irradiated mice. The FST significantly decreased the catalase (CAT) activity and total glutathione (t-GSH) content in the brain and kidney, respectively. The superoxide dismutase (SOD) activity and t-GSH content in the liver of the 3.0 mGy/h irradiated mice were significantly lower than those of the non-FST-treated mice. The CAT activity in the lungs of mice exposed to 3.0 mGy/h γ-irradiation was higher than that of non-FST treated mice and mice treated with FST. However, no significant differences were observed in the levels of these antioxidant markers between the sham and irradiated groups except for the CAT activity in lungs. These findings suggest that the effects of low-dose-rate and low-dose γ-irradiation on FST are highly organ-dependent.


Modelling farmer choices for water security measures in the Litani river basin in Lebanon.
Lebanon is facing an increasing water supply deficit due to the increasing demand for freshwater, decreasing surface and groundwater resources and malfunctioning water governance structures. Technological and policy changes are needed to alleviate the impact of water scarcity and secure water in the future. This paper investigates farmers' preferences and willingness to pay (WTP) in a choice experiment for a series of water saving measures at plot and irrigation district level, including more timely information of water delivery. These measures are expected to strengthen water security and use water more efficiently. Farmers are willing to pay higher water prices of $0.32/m3 and $0.22/m3 to support the implementation of water saving measures at plot level and the installation of water metering devices across the irrigation district, respectively. They are not willing to pay extra for obtaining information related to their water delivery earlier in time if this means that they will also have to pay earlier in the year for the water. Farmers with higher income and education levels who decide on their cropping pattern based on expected rainfall data are more interested in taking action than farmers whose cropping decisions are primarily based on last year's sales prices. The study shows that when aiming to design more effective sustainable water management strategies, accounting for farmers' needs and preferences, their age also has to be considered: younger farmers (<40 years) are on average more interested in and willing to pay more for new water saving measures than older farmers (>40 years).


Stress-induced alternative splice forms of MDM2 and MDMX modulate the p53-pathway in distinct ways.
MDM2 and MDMX are the chief negative regulators of the tumor-suppressor protein p53 and are essential for maintaining homeostasis within the cell. In response to genotoxic stress and also in several cancer types, MDM2 and MDMX are alternatively spliced. The splice variants MDM2-ALT1 and MDMX-ALT2 lack the p53-binding domain and are incapable of negatively regulating p53. However, they retain the RING domain that facilitates dimerization of the full-length MDM proteins. Concordantly, MDM2-ALT1 has been shown to lead to the stabilization of p53 through its interaction with and inactivation of full-length MDM2. The impact of MDM2-ALT1 expression on the p53 pathway and the nature of its interaction with MDMX remain unclear. Also, the role of the architecturally similar MDMX-ALT2 and its influence of the MDM2-MDMX-p53 axis are yet to be elucidated. We show here that MDM2-ALT1 is capable of binding full-length MDMX as well as full-length MDM2. Additionally, we demonstrate that MDMX-ALT2 is able to dimerize with both full-length MDMX and MDM2 and that the expression of MDM2-ALT1 and MDMX-ALT2 leads to the upregulation of p53 protein, and also of its downstream target p21. Moreover, MDM2-ALT1 expression causes cell cycle arrest in the G1 phase in a p53 and p21 dependent manner, which is consistent with the increased levels of p21. Finally we present evidence that MDM2-ALT1 and MDMX-ALT2 expression can activate subtly distinct subsets of p53-transcriptional targets implying that these splice variants can modulate the p53 tumor suppressor pathway in unique ways. In summary, our study shows that the stress-inducible alternative splice forms MDM2-ALT1 and MDMX-ALT2 are important modifiers of the p53 pathway and present a potential mechanism to tailor the p53-mediated cellular stress response.


Genome-wide association studies identify CHRNA5/3 and HTR4 in the development of airflow obstruction.
Genome-wide association studies (GWAS) have identified loci influencing lung function, but fewer genes influencing chronic obstructive pulmonary disease (COPD) are known. Perform meta-analyses of GWAS for airflow obstruction, a key pathophysiologic characteristic of COPD assessed by spirometry, in population-based cohorts examining all participants, ever smokers, never smokers, asthma-free participants, and more severe cases. Fifteen cohorts were studied for discovery (3,368 affected; 29,507 unaffected), and a population-based family study and a meta-analysis of case-control studies were used for replication and regional follow-up (3,837 cases; 4,479 control subjects). Airflow obstruction was defined as FEV(1) and its ratio to FVC (FEV(1)/FVC) both less than their respective lower limits of normal as determined by published reference equations. The discovery meta-analyses identified one region on chromosome 15q25.1 meeting genome-wide significance in ever smokers that includes AGPHD1, IREB2, and CHRNA5/CHRNA3 genes. The region was also modestly associated among never smokers. Gene expression studies confirmed the presence of CHRNA5/3 in lung, airway smooth muscle, and bronchial epithelial cells. A single-nucleotide polymorphism in HTR4, a gene previously related to FEV(1)/FVC, achieved genome-wide statistical significance in combined meta-analysis. Top single-nucleotide polymorphisms in ADAM19, RARB, PPAP2B, and ADAMTS19 were nominally replicated in the COPD meta-analysis. These results suggest an important role for the CHRNA5/3 region as a genetic risk factor for airflow obstruction that may be independent of smoking and implicate the HTR4 gene in the etiology of airflow obstruction.


Fermented foods, microbiota, and mental health: ancient practice meets nutritional psychiatry.
The purposeful application of fermentation in food and beverage preparation, as a means to provide palatability, nutritional value, preservative, and medicinal properties, is an ancient practice. Fermented foods and beverages continue to make a significant contribution to the overall patterns of traditional dietary practices. As our knowledge of the human microbiome increases, including its connection to mental health (for example, anxiety and depression), it is becoming increasingly clear that there are untold connections between our resident microbes and many aspects of physiology. Of relevance to this research are new findings concerning the ways in which fermentation alters dietary items pre-consumption, and in turn, the ways in which fermentation-enriched chemicals (for example, lactoferrin, bioactive peptides) and newly formed phytochemicals (for example, unique flavonoids) may act upon our own intestinal microbiota profile. Here, we argue that the consumption of fermented foods may be particularly relevant to the emerging research linking traditional dietary practices and positive mental health. The extent to which traditional dietary items may mitigate inflammation and oxidative stress may be controlled, at least to some degree, by microbiota. It is our contention that properly controlled fermentation may often amplify the specific nutrient and phytochemical content of foods, the ultimate value of which may associated with mental health; furthermore, we also argue that the microbes (for example, Lactobacillus and Bifidobacteria species) associated with fermented foods may also influence brain health via direct and indirect pathways.


Prognostic influence on survival of increased ornithine decarboxylase activity in human breast cancer.
Although considerable experimental evidence suggests an important role of polyamines in breast cancer biology, compelling supportive data in patients are lacking. To address this issue, we measured ornithine decarboxylase (ODC), S-adenosylmethionine decarboxylase, and spermidine/spermine acetyltransferase (the three key polyamine metabolic enzymes) in a cohort of 50 primary human breast cancers and related their levels of activity to disease-free survival and overall survival. The major finding of our study was that ODC activity level was a negative independent prognostic factor for both end points. With regard to overall survival, the adverse influence of ODC expression was superior even to that provided by the number of positive nodes. Furthermore, the statistical significance of the ODC effect on survival was enhanced when breast cancer-specific mortality was included in the analysis as opposed to death from any cause. In addition, high tumor ODC activity may predict a shorter time from recurrence to death, although this effect was of only borderline statistical significance. In summary, these results provide the first concrete evidence supporting the prognostic role of ODC in human breast cancer.


Assessment of dysarthria with Frenchay dysarthria assessment (FDA-2) in patients with Duchenne muscular dystrophy.
The purpose of this study was to test the psychometric properties of the Japanese version of Frenchay Dysarthria Assessment (FDA-2) and to use this tool to describe the features of speech in patients with Duchenne muscular dystrophy (DMD). The Japanese version of FDA-2 was culturally adapted, and reliability and validity were examined in 22 and 50 patients, respectively. The Japanese version of FDA-2 was administered to 51 patients with DMD. Multiple regression analysis was performed to identify factors related to FDA-2 scores. Inter-/intra-rater reliabilities (ICCs) and internal consistency (Cronbach's α) for total scores were 0.76, 0.97, and 0.94 respectively. For construct validity, two-way ANOVA showed a significant interaction between the disorders and FDA-2 sections (p < 0.05). In DMD patients, the item of tongue at rest was most severely affected, reflecting tongue hypertrophy. Multiple regression analysis identified age, swallowing status, and ventilator use as significantly related. The results showed that the Japanese version of FDA-2 has satisfactory reliability and validity. The present study demonstrated the features of dysarthria and related factors in patients with DMD.Implications for rehabilitationIn Duchenne muscular dystrophy (DMD), an absent or defective dystrophin protein causes progressive weakness of respiratory and oropharyngeal muscles, both of which are crucial contributors to speech production.This study shows that the Japanese version of FDA-2 has satisfactory reliability and validity compared to original version.The Japanese version of FDA-2 characterizes dysarthria in patients with DMD in this cohort.


Disassembling archeology, reassembling the modern world.
This article provides a substantive discussion of the relevance of the history of archeology to the history of science. At the same time, the article introduces the papers contained in this special issue as exemplars of this relevance. To make its case, the article moves through various themes in the history of archeology that overlap with key issues in the history of science. The article discusses the role and tension of regimes of science in antiquarian and archeological practices, and also considers issues of scale and place, particularly in relation to the field. Additionally, the piece attends to issues of professionalization and the constitution of an archeological public, at the same time as discussing issues of empire, colonialism, and the circulation of knowledge. Meanwhile, enriching discussions within and beyond the history of science, the article discusses the history of archeology and its relationship with museums, collecting, and material culture and materiality. Finally, the piece discusses the relationship of the history of archeology with wider discussions about scientific ethics. In conclusion, the article questions whether we should speak of 'the history of archeology' at all.


Diet drives convergence in gut microbiome functions across mammalian phylogeny and within humans.
Coevolution of mammals and their gut microbiota has profoundly affected their radiation into myriad habitats. We used shotgun sequencing of microbial community DNA and targeted sequencing of bacterial 16S ribosomal RNA genes to gain an understanding of how microbial communities adapt to extremes of diet. We sampled fecal DNA from 33 mammalian species and 18 humans who kept detailed diet records, and we found that the adaptation of the microbiota to diet is similar across different mammalian lineages. Functional repertoires of microbiome genes, such as those encoding carbohydrate-active enzymes and proteases, can be predicted from bacterial species assemblages. These results illustrate the value of characterizing vertebrate gut microbiomes to understand host evolutionary histories at a supraorganismal level.


The Oxford Positive Self Scale: psychometric development of an assessment of cognitions associated with psychological well-being.
Developing, elaborating, and consolidating positive views of the self is a plausible route to increased psychological well-being. We set out to provide an assessment of positive self-beliefs that could be used in research and clinical practice. A non-probability online survey was conducted with 2500 UK adults, quota sampled to match the population for age, gender, ethnicity, income, and region. Exploratory factor analysis of a 94-item pool - generated with guidance from people with lived experience of mental health difficulties - was conducted to develop the Oxford Positive Self Scale (OxPos). The item pool was further reduced using regularised structural equation modelling (SEM) before confirmatory factor analysis. Optimal cut-off scores were developed using receiver operating characteristic curves. Additional validations were carried out with two further general population cohorts (n = 1399; n = 1693). A 24-item scale was developed with an excellent model fit [robust χ2 = 995.676; df = 246; CFI = 0.956; TLI = 0.951; RMSEA = 0.049 (0.047, 0.052); SRMR = 0.031]. The scale comprises four factors: mastery; strength; enjoyment; and character. SEM indicated that the scale explains 68.6% of variance in psychological well-being. The OxPos score was negatively correlated with depression (r = -0.49), anxious avoidance (r = -0.34), paranoia (r = -0.23), hallucinations (r = -0.20), and negative self-beliefs (r = -0.50), and positively correlated with psychological well-being (r = 0.79), self-esteem (r = 0.67), and positive social comparison (r = 0.72). Internal reliability and test-retest reliability were excellent. Cut-offs by age and gender were generated. A short-form was developed, explaining 96% of the full-scale variance. The new open access scale provides a psychometrically robust assessment of positive cognitions that are strongly connected to psychological well-being.


Moving Along: In biomechanics, rehabilitation engineering, and movement analysis, Italian researchers are making great strides.
In Italy, biomechanics research and the analysis of human and animal movement have had a very long history, beginning with the exceptional pioneering work of Leonardo da Vinci. In 1489, da Vinci began investigating human anatomy, including an examination of human tendons, muscles, and the skeletal system. He continued this line of inquiry later in life, identifying what he called "the four powers--movement, weight, force, and percussion"--and how he thought they worked in the human body. His approach, by the way, was very modern--analyzing nature through anatomy, developing models for interpretation, and transferring this knowledge to bio-inspired machines.


The clinical significance of positive blood cultures: a comprehensive analysis of 500 episodes of bacteremia and fungemia in adults. I. Laboratory and epidemiologic observations.
Five hundred episodes of septicemia were reviewed, with emphasis on laboratory and epidemiologic findings. The isolation of facultative and anaerobic gram-negative bacilli, fungi, and gram-positive cocci (except viridans streptococci and Staphylococcus epidermidis) almost always indicated true bacteremia, whereas the isolation of aerobic and anaerobic gram-positive bacilli, including Clostridium species, often represented contamination. More than 99% of all episodes were detected when two samples of blood (a total of 30 ml) were cultured. The five most common isolates were Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniae, Klebsiella pneumoniae, and Pseudomonas aeruginosa. The incidence of septicemia was highest among medical patients and lowest among obstetric-gynecologic patients. Two-thirds of all episodes were nosocomial; S. aureus, enterococci, facultative gram-negative bacilli, and fungi were especially common nosocomial pathogens. The microorganisms isolated varied with the hospital service; polymicrobial episodes were especially common among surgical patients and transplant recipients. The most common sources of bacteremia were the respiratory, genitourinary, and gastrointestinal tracts; however, the source was unknown in nearly one-third of episodes. Microorganisms causing septicemia in neutropenic and nonneutropenic patients were not different; however, polymicrobial infections were more frequent in the presence of neutropenia. After antimicrobial susceptibility data became available, therapy was appropriate greater than 90% of the time.


Intradiscal injection of simvastatin results in radiologic, histologic, and genetic evidence of disc regeneration in a rat model of degenerative disc disease.
A large percentage of back pain can be attributed to degeneration of the intervertebral disc (IVD). Bone morphogenetic protein 2 (BMP-2) is known to play an important role in chondrogenesis of the IVD. Simvastatin is known to upregulate expression of BMP-2. Thus, we hypothesized that intradiscal injection of simvastatin in a rat model of degenerative disc disease (DDD) would result in retardation of DDD. The purpose of the present study was to develop a novel conservative treatment for DDD and related discogenic back pain. The setting of this study is the laboratory investigation. Disc injury was induced in 272 rats via 21-ga needle puncture. After 6 weeks, injured discs were treated with simvastatin in a saline or hydrogel carrier. Rats were sacrificed at predetermined time points. Outcome measures assessed were radiologic, histologic, and genetic. Radiologically, the magnetic resonance imaging (MRI) index (number of pixels multiplied by the corresponding image densities) was determined. Histologically, disc spaces were read by three blinded scorers using a previously described histologic grading scale. Genetically, nuclei pulposi were harvested, and polymerase chain reaction was run to determine relative levels of aggrecan, collagen type II, and BMP-2 gene expression. Radiologically, discs treated with 5 mg/mL of simvastatin in hydrogel or saline demonstrated MRI indices that were normal through 8 weeks after treatment, although this was more sustained when delivered in hydrogel. Histologically, discs treated with 5 mg/mL of simvastatin in hydrogel demonstrated improved grades compared with discs treated at higher doses. Genetically, discs treated with 5 mg/mL of simvastatin in hydrogel demonstrated higher gene expression of aggrecan and collagen type II than control. Degenerate discs treated with 5 mg/mL of simvastatin in a hydrogel carrier demonstrated radiographic and histologic features resembling normal noninjured IVDs. In addition, the gene expression of aggrecan and collagen type II (important constituents of the IVD extracellular matrix) was upregulated in treated discs. Injection of simvastatin into degenerate IVDs may result in retardation of disc degeneration and represents a promising investigational therapy for conservative treatment of DDD.