Management of test results in family medicine offices.
We wanted to explore test results management systems in family medicine offices and to delineate the components of quality in results management. Using a multimethod protocol, we intensively studied 4 purposefully chosen family medicine offices using observations, interviews, and surveys. Data analysis consisted of iterative qualitative analysis, descriptive frequencies, and individual case studies, followed by a comparative case analysis. We assessed the quality of results management at each practice by both the presence of and adherence to systemwide practices for each results management step, as well as outcomes from chart reviews, patient surveys, and interview and observation notes. We found variability between offices in how they performed the tasks for each of the specific steps of results management. No office consistently had or adhered to office-wide results management practices, and only 2 offices had written protocols or procedures for any results management steps. Whereas most patients surveyed acknowledged receiving their test results (87% to 100%), a far smaller proportion of patient charts documented patient notification (58% to 85%), clinician response to the result (47% to 84%), and follow-up for abnormal results (28% to 55%). We found 2 themes that emerged as factors of importance in assessing test results management quality: safety awareness-a leadership focus and communication that occurs around quality and safety, teamwork in the office, and the presence of appropriate policies and procedures; and technological adoption-the presence of an electronic health record, digital connections between the office and testing facilities, use of technology to facilitate patient communication, and the presence of forcing functions (built-in safeguards and requirements). Understanding the components of safety awareness and technological adoption can assist family medicine offices in evaluating their own results management processes and help them design systems that can lead to higher quality care.


Perception, Price and Preference: Consumption and Protection of Wild Animals Used in Traditional Medicine.
A wide array of wildlife species, including many animals, are used in traditional medicines across many medicinal systems, including in Traditional Chinese Medicine (TCM). Due to over-exploitation and habitat loss, the populations of many animals commonly used in TCM have declined and are unable to meet market demand. A number of measures have been taken to try to reduce the impact that this large and growing market for TCM may have on wild animal species. Consumer preferences and behavior are known to play an important role in the consumption and protection of wild animals used in traditional medicine, and thus are likely to be an important factor in the success of many of these mechanisms--particularly given the significant percentage of TCMs that are over-the-counter products (access to which is not mediated by practitioners). In this study we conducted questionnaires and designed stated preference experiments embodying different simulation scenarios using a random sample of the population in Beijing to elicit individuals' knowledge, perceptions and preferences toward wild or farmed animal materials and their substitutes used in traditional Chinese medicine. We found that respondents had a stated preference for wild materials over farm-raised and other alternatives because they believe that the effectiveness of wild-sourced materials is more credible than that of other sources. However, we also found that, although respondents used TCM products, they had a poor understanding of the function or composition of either traditional Chinese medicines or proprietary Chinese medicines (PCM), and paid little attention to the composition of products when making purchasing decisions. Furthermore, awareness of the need for species protection, or "conservation consciousness" was found to play an important role in willingness to accept substitutions for wild animal materials, while traditional animal medicinal materials (TAMs) derived from well-known endangered species, such as bear bile and tiger bone, show relatively higher substitutability. These results suggest that there is still hope for conservation measures which seek to promote a transition to farmed animal, plant and synthetic ingredients and provide clear directions for future social marketing, education and engagement efforts.


Physician-directed primary care intervention to reduce risk factors for type 2 diabetes in high-risk youth.
The incidence of type 2 diabetes in youth is increasing at an alarming rate. The purpose of this pilot study was to determine whether a nutrition and physical activity intervention in an urban primary care office is feasible and effective in decreasing risk factors for type 2 diabetes in high-risk youth. A one-group pretest/post-test design was used. Participants were recruited from existing patients in a primary care facility serving low-income children. Inclusion criteria included body mass index (BMI) over the 85th percentile for age and a fasting glucose-insulin ratio (FGIR) less than 6. Thirty-six African-American patients, 9 males, 27 females, average age 12.4 years (range, 8-18) participated in a 12-week nutrition and physical activity program. Measurements included fasting glucose, insulin, FGIR, lipid profile, blood pressure, and BMI. BMI and laboratory values were tested for significant differences before and after intervention using paired t-tests. A P-value of <.05 was considered statistically significant. On average, patients attended 8.3 of 24 physical activity sessions, 2 of 3 nutrition sessions and 1.5 of 3 planned clinical sessions. Twenty-six of 36 patients completed follow-up laboratory tests. Mean FGIR improved significantly from baseline (3.6 +/- 1.2 to 4.6 +/- 2.8; P = .043). A nutrition and physical activity intervention for overweight children can be conducted in an urban primary care setting and may decrease laboratory evidence of insulin resistance, a risk factor for type 2 diabetes. Making the program accessible by public transit and scheduling the sessions at convenient times were important factors.


The Botrytis cinerea Xylanase BcXyl1 Modulates Plant Immunity.
Botrytis cinerea is one of the most notorious pathogenic species that causes serious plant diseases and substantial losses in agriculture throughout the world. We identified BcXyl1 from B. cinerea that exhibited xylanase activity. Expression of the BcXyl1 gene was strongly induced in B. cinerea infecting Nicotiana benthamiana and tomato plants, and BcXyl1 deletion strains severely compromised the virulence of B. cinerea. BcXyl1 induced strong cell death in several plants, and cell death activity of BcXyl1 was independent of its xylanase activity. Purified BcXyl1 triggered typically PAMP-triggered immunity (PTI) responses and conferred resistance to B. cinerea and TMV in tobacco and tomato plants. A 26-amino acid peptide of BcXyl1 was sufficient for elicitor function. Furthermore, the BcXyl1 death-inducing signal was mediated by the plant LRR receptor-like kinases (RLKs) BAK1 and SOBIR1. Our data suggested that BcXyl1 contributed to B. cinerea virulence and induced plant defense responses.


Genomic Fusions in Pigmented Spindle Cell Nevus of Reed.
Recent molecular studies of spitzoid neoplasms have identified mutually exclusive kinase fusions involving ROS1, ALK, RET, BRAF, NTRK1, MET, and NTRK3 as early initiating genomic events. Pigmented spindle cell nevus (PSCN) of Reed is a morphologic variant of Spitz and may be very diagnostically challenging, having histologic features concerning for melanoma. Their occurrence in younger patients, lack of association to sun exposure, and rapid early growth phase similar to Spitz nevi suggest fusions may also play a significant role in these lesions. However, to date, there is little data in the literature focused on the molecular characterization of PSCN of Reed with next-generation sequencing. We analyzed a total of 129 melanocytic neoplasms with RNA sequencing including 67 spitzoid neoplasms (10 Spitz nevi, 44 atypical Spitz tumors, 13 spitzoid melanomas) and 23 PSCN of Reed. Although only 2 of 67 (3.0%) of spitzoid lesions had NTRK3 fusions, 13 of 23 (57%) of PSCN of Reed harbored NTRK3 fusions with 5' partners ETV6 (12p13) in 2 cases and MYO5A (15q21) in 11 cases. NTRK3 fusions were confirmed with a fluorescent in situ hybridization break-apart probe. The presence of a NTRK3 fusion correlated with younger age (P=0.021) and adnexal extension (P=0.001). Other minor fusions identified in PSCN of Reed included MYO5A-MERTK (2), MYO5A-ROS1, MYO5A-RET, and ETV6-PITX3 leading to a total of 78% with fusions. Our study suggests that the majority of PSCN of Reed are the result of genomic fusions, and the most frequent and characteristic genomic aberration is an NTRK3 fusion.


Reduced secretion of virions and hepatitis B virus (HBV) surface antigen of a naturally occurring HBV variant correlates with the accumulation of the small S envelope protein in the endoplasmic reticulum and Golgi apparatus.
We identified two novel naturally occurring mutations (W74L and L77R) in the small S envelope protein of hepatitis B virus (HBV). Mutation L77R alone resulted in >10-fold-reduced secretion of virions. In addition, the 2.8-fold reduction of the extracellular HBV surface antigen (HBsAg) of mutant L77R from transfected Huh7 cells appeared to be correlated with a 1.7-fold reduction of intracellular HBsAg, as measured by enzyme-linked immunosorbent assay (ELISA). Surprisingly, opposite to the ELISA results, Western blot analysis revealed a near-10-fold-increased level of the intracellular mutant small S envelope protein. The discrepancy between ELISA and Western blot data was due to significant accumulation of the mutant L77R HBsAg in the intracellular pellet fraction. In contrast to HBsAg, the secretion of HBeAg was normal in L77R-transfected cells. The wild-type HBsAg was usually more diffuse and evenly distributed in the cytoplasm, often outside the perinuclear endoplasmic reticulum (ER) and Golgi apparatus, as observed by immunofluorescence assay. In contrast, the L77R mutant HBsAg tends to be highly restricted within the ER and Golgi, often accumulated in the Golgi compartments distal from the nucleus. The almost exclusive retention in the ER-Golgi of L77R HBsAg was similar to what was observed when the large envelope protein was overexpressed. These multiple aberrant phenotypes of mutant L77R can be corrected by a second naturally occurring S envelope mutation, W74L. Despite the accumulation of L77R HBsAg in ER-Golgi of transfected Huh7 cells, we detected no increase in Grp78 mRNA and proteins, which are common markers for ER stress response.


Spatially augmented LPboosting for AD classification with evaluations on the ADNI dataset.
Structural and functional brain images are playing an important role in helping us understand the changes associated with neurological disorders such as Alzheimer's disease (AD). Recent efforts have now started investigating their utility for diagnosis purposes. This line of research has shown promising results where methods from machine learning (such as Support Vector Machines) have been used to identify AD-related patterns from images, for use in diagnosing new individual subjects. In this paper, we propose a new framework for AD classification which makes use of the Linear Program (LP) boosting with novel additional regularization based on spatial "smoothness" in 3D image coordinate spaces. The algorithm formalizes the expectation that since the examples for training the classifier are images, the voxels eventually selected for specifying the decision boundary must constitute spatially contiguous chunks, i.e., "regions" must be preferred over isolated voxels. This prior belief turns out to be useful for significantly reducing the space of possible classifiers and leads to substantial benefits in generalization. In our method, the requirement of spatial contiguity (of selected discriminating voxels) is incorporated within the optimization framework directly. Other methods have made use of similar biases as a pre- or post-processing step, however, our model incorporates this emphasis on spatial smoothness directly into the learning step. We report on extensive evaluations of our algorithm on MR and FDG-PET images from the Alzheimer's Disease Neuroimaging Initiative (ADNI) dataset, and discuss the relationship of the classification output with the clinical and cognitive biomarker data available within ADNI.


Comparative photoactivity and antibacterial properties of C60 fullerenes and titanium dioxide nanoparticles.
The production of reactive oxygen species (ROS) by aqueous suspensions of fullerenes and nano-TiO2 (Degussa P25) was measured both in ultrapure water and in minimal Davis (MD) microbial growth medium. Fullerol (hydroxylated C60) produced singlet oxygen (1O2) in ultrapure water and both 1O2 and superoxide (O2-*) in MD medium, but no hydroxyl radicals (OH*) were detected in either case. PVP/C60 (C60 encapsulated with poly(N-vinylpyrrolidone)) was more efficient than fullerol in generating singlet oxygen and superoxide. However, two other aggregates of C60, namely THF/nC60 (prepared with tetrahydofuran as transitional solvent) and aqu/nC60 (prepared by vigorous stirring of C60 powder in water), were not photoactive. Nano-TiO2 (also present as aggregates) primarily produced hydroxyl radicals in pure water and superoxide in MD medium. Bacterial (Escherichia coli) toxicity tests suggest that, unlike nano-TiO2 which was exclusively phototoxic, the antibacterial activity of fullerene suspensions was linked to ROS production. Nano-TiO2 may be more efficient for water treatment involving UV or solar energy, to enhance contaminant oxidation and perhaps for disinfection. However, fullerol and PVP/ C60 may be useful as water treatment agents targeting specific pollutants or microorganisms that are more sensitive to either superoxide or singlet oxygen.


Mnemonic strategy training partially restores hippocampal activity in patients with mild cognitive impairment.
Learning and memory deficits typify patients with mild cognitive impairment (MCI) and are generally attributed to medial temporal lobe dysfunction. Although the hippocampus is perhaps the most commonly studied neuroanatomical structure in these patients, there have been few attempts to identify rehabilitative interventions that facilitate its functioning. Here, we present results from a randomized, controlled, single-blind study in which patients with MCI and healthy elderly controls (HEC) were randomized to either three sessions of mnemonic strategy training (MS) or a matched-exposure control group (XP). All participants underwent pre- and posttraining fMRI scanning as they encoded and retrieved object-location associations. For the current report, fMRI analyses were restricted to the hippocampus, as defined anatomically. Before training, MCI patients showed reduced hippocampal activity during both encoding and retrieval, relative to HEC. Following training, the MCI MS group demonstrated increased activity during both encoding and retrieval. There were significant differences between the MCI MS and MCI XP groups during retrieval, especially within the right hippocampus. Thus, MS facilitated hippocampal functioning in a partially restorative manner. We conclude that cognitive rehabilitation techniques may help mitigate hippocampal dysfunction in MCI patients.


What is the contribution of physician associates in hospital care in England? A mixed methods, multiple case study.
To investigate the deployment of physician associates (PAs); the factors supporting and inhibiting their employment and their contribution and impact on patients' experience and outcomes and the organisation of services. Mixed methods within a case study design, using interviews, observations, work diaries and documentary analysis. Six acute care hospitals in three regions of England in 2016-2017. 43 PAs, 77 other health professionals, 28 managers, 28 patients and relatives. A key influencing factor supporting the employment of PAs in all settings was a shortage of doctors. PAs were found to be acceptable, appropriate and safe members of the medical/surgical teams by the majority of doctors, managers and nurses. They were mainly deployed to undertake inpatient ward work in the medical/surgical team during core weekday hours. They were reported to positively contribute to: continuity within their medical/surgical team, patient experience and flow, inducting new junior doctors, supporting the medical/surgical teams' workload, which released doctors for more complex patients and their training. The lack of regulation and attendant lack of authority to prescribe was seen as a problem in many but not all specialties. The contribution of PAs to productivity and patient outcomes was not quantifiable separately from other members of the team and wider service organisation. Patients and relatives described PAs positively but most did not understand who and what a PA was, often mistaking them for doctors. This study offers new insights concerning the deployment and contribution of PAs in medical and surgical specialties in English hospitals. PAs provided a flexible addition to the secondary care workforce without drawing from existing professions. Their utility in the hospital setting is unlikely to be completely realised without the appropriate level of regulation and authority to prescribe medicines and order ionising radiation within their scope of practice.


Cytosolic group IVA phospholipase A2 inhibitors, AVX001 and AVX002, ameliorate collagen-induced arthritis.
Cytosolic phospholipase A2 group IVA (cPLA2α)-deficient mice are resistant to collagen-induced arthritis, suggesting that cPLA2α is an important therapeutic target. Here, the anti-inflammatory effects of the AVX001 and AVX002 cPLA2α inhibitors were investigated. In vitro enzyme activity was assessed by a modified Dole assay. Effects on inhibiting IL-1β-induced release of arachidonic acid (AA) and prostaglandin E2 (PGE2) were measured using SW982 synoviocyte cells. In vivo effects were studied in prophylactic and therapetic murine collagen-induced arthritis models and compared to methotrexate (MTX) and Enbrel, commonly used anti-rheumatic drugs. The in vivo response to treatment was evaluated in terms of the arthritis index (AI), histopathology scores and by plasma levels of PGE2 following 14 and 21 days of treatment. Both cPLA2α inhibitors are potent inhibitors of cPLA2α in vitro. In synoviocytes, AVX001 and AVX002 reduce, but do not block, release of AA or PGE2 synthesis. In both CIA models, the AI and progression of arthritis were significantly lower in the mice treated with AVX001, AVX002, Enbrel and MTX than in non- treated mice. Several histopathology parameters of joint damage were found to be significantly reduced by AVX001 and AVX002 in both prophylactic and therapeutic study modes; namely articular cavity and peripheral tissue inflammatory cell infiltration; capillary and synovial hyperplasia; articular cartilage surface damage; and periostal and endochondral ossification. In comparison, MTX did not significantly improve any histopathology parameters and Enbrel only improved ossification. Finally, as a biomarker of inflammation and as an indication that AVX001 and AVX002 blocked the cPLA2α target, we determined that plasma levels of PGE2 were significantly reduced in response to the AVX inhibitors and MTX, but not Enbrel. AVX001 and AVX002 display potent anti-inflammatory activity and disease-modifying properties in cellular and in vivo models. The in vivo effects of AVX001 and AVX002 were comparable to, or superior, to those of MTX and Enbrel. Taken together, this study suggests that cPLA2α inhibitors AVX001 and AVX002 are promising small molecule disease-modifying anti-rheumatic therapies.


Adding unregulated nursing support workers to ward staffing: Exploration of a natural experiment.
To explore the impact of an initiative to add unregulated nursing support workers to wards in acute care hospitals. Adding nursing support workers to existing nurse staffing may be one solution to reduce nursing workloads and improve outcomes. However, the effects of this addition on nurse, patient and system outcomes are not well documented. In one state of Australia, a trial deployment of nursing support workers to wards across the public health system provided opportunity for the exploration of their impact in a natural, real-world, environment. Cross-sectional study. A sample of five wards where nursing support workers had been added matched to a group of five wards where there were no nursing support workers. Data were collected via patient survey (n = 141) and nurse survey (n = 154). Analysis was comparative with regression models constructed for the different ward types. Nursing leadership, staffing and resources, and nurse experience were linked to outcomes on both ward types. Instability was a significant predictor of reduced quality of care and increased turnover intention on wards where support workers were added. Adding nursing support workers to ward staffing did not lead to improvements in patient care. Findings suggest that staffing a nursing ward is a complex activity and that a simple approach to staffing is unlikely to be successful. Future research should explore the process of implementation and the conditions under which this strategy is likely to be successful. Ward-level factors are key in making appropriate staffing and skill mix choices to limit instability and to consequently avoid negative patient, staff and system outcomes. Consideration of the ward context, alongside effective delegation processes and integration into the care team are imperative when adding nursing support workers.


Comparison of contracture, adhesion, tissue ingrowth, and histologic response characteristics of permanent and absorbable barrier meshes in a porcine model of laparoscopic ventral hernia repair.
The objective of this study was to determine the mesh contracture, adhesion, tissue ingrowth, and histologic characteristics of a novel absorbable barrier mesh (Ventrio™ ST Hernia Patch) compared to existing permanent (Ventrio™ Hernia Patch) and absorbable barrier meshes (Sepramesh™ IP Composite and PROCEED™ Surgical Mesh). Standard laparoscopic technique was utilized to bilaterally implant meshes in 20 female Yorkshire pigs (n = 5 pigs/group). Meshes were fixated to the intact peritoneum with SorbaFix™ absorbable fixation devices. Mesh contracture, adhesion coverage, and adhesion tenacity were evaluated after 4 weeks. T-Peel testing and hematoxylin and eosin (H&E) staining were utilized to assess tissue ingrowth and host response. A significantly greater percent area contracture was demonstrated for PROCEED™ (26.9%) compared to Ventrio™ ST (8.8%), Ventrio™ (14.5%) and Sepramesh™ (9.2%). Ventrio™ ST demonstrated similar adhesion area, tenacity, and tissue ingrowth compared to all other meshes. Histological scoring revealed a comparable host inflammatory response for all meshes, with a few exceptions. A greater number of giant cells were observed in Ventrio™ ST and Sepramesh™ near the multifilament polyglycolic acid (PGA) fibers; a greater number of macrophages were observed in PROCEED™ compared to Ventrio™; and a greater number of neutrophils were observed in PROCEED™, compared to Sepramesh™ (P < 0.05). Focal areas of hemorrhage were also observed on the visceral surface of PROCEED™. Ventrio™ ST Hernia Patch demonstrated comparable contracture, adhesion, tissue ingrowth, and histologic characteristics compared to existing permanent and absorbable barrier meshes. Host inflammatory and fibrotic responses for all four meshes were minimal and representative of a biocompatible response.


IL-15 has stimulatory activity for the induction of B cell proliferation and differentiation.
The identification and cloning of the novel cytokine IL-15 were recently described. IL-15 is produced by a wide range of cell types, with the highest levels of IL-15 mRNA being detected in epithelial lines, monocytes, muscle, and placenta. Although it has no sequence identity with IL-2, IL-15 shares many of the T cell-stimulatory activities described for IL-2. We have examined IL-15 for its ability to stimulate B cells and have compared its activity with that of IL-2. IL-15 costimulates proliferation of B cells activated with immobilized anti-human IgM or phorbol ester, but has no stimulatory effect on resting B cells. In combination with recombinant CD40L, IL-15 is a potent inducer of polyclonal IgM, IgG1, and IgA secretion, but does not cause production of IgG4 or IgE. The activity of IL-15 in B cell proliferation and differentiation assays is comparable with that of IL-2. Studies that used neutralizing Abs have demonstrated that, for signal transduction in B cells, IL-15 uses the beta-chain of the IL-2R complex but, unlike IL-2, does not require the alpha-chain. IL-2 is required for the generation of a human primary Ag-specific in vitro response using sheep erythrocytes as Ag. Of all cytokines examined, only IL-15 has the capacity to replace IL-2 in this system, although only partially. In summary, IL-15 has comparable activity with IL-2 for the induction of B cell proliferation and differentiation and uses at least some of the components of the IL-2R complex to mediate its effects.


Biases in Drosophila melanogaster protein trap screens.
The ability to localise or follow endogenous proteins in real time in vivo is of tremendous utility for cell biology or systems biology studies. Protein trap screens utilise the random genomic insertion of a transposon-borne artificial reporter exon (e.g. encoding the green fluorescent protein, GFP) into an intron of an endogenous gene to generate a fluorescent fusion protein. Despite recent efforts aimed at achieving comprehensive coverage of the genes encoded in the Drosophila genome, the repertoire of genes that yield protein traps is still small. We analysed the collection of available protein trap lines in Drosophila melanogaster and identified potential biases that are likely to restrict genome coverage in protein trap screens. The protein trap screens investigated here primarily used P-element vectors and thus exhibit some of the same positional biases associated with this transposon that are evident from the comprehensive Drosophila Gene Disruption Project. We further found that protein trap target genes usually exhibit broad and persistent expression during embryonic development, which is likely to facilitate better detection. In addition, we investigated the likely influence of the GFP exon on host protein structure and found that protein trap insertions have a significant bias for exon-exon boundaries that encode disordered protein regions. 38.8% of GFP insertions land in disordered protein regions compared with only 23.4% in the case of non-trapping P-element insertions landing in coding sequence introns (p < 10(-4)). Interestingly, even in cases where protein domains are predicted, protein trap insertions frequently occur in regions encoding surface exposed areas that are likely to be functionally neutral. Considering the various biases observed, we predict that less than one third of intron-containing genes are likely to be amenable to trapping by the existing methods. Our analyses suggest that the utility of P-element vectors for protein trap screens has largely been exhausted, and that approximately 2,800 genes may still be amenable using piggyBac vectors. Thus protein trap strategies based on current approaches are unlikely to offer true genome-wide coverage. We suggest that either transposons with reduced insertion bias or recombineering-based targeting techniques will be required for comprehensive genome coverage in Drosophila.


How Effective Is Pulse Arrival Time for Evaluating Blood Pressure? Challenges and Recommendations from a Study Using the MIMIC Database.
Cardiovascular disease (CVD) is the number one cause of non-infectious morbidity and mortality in the world. The detection, measurement, and management of high blood pressure play an essential role in the prevention and control of CVDs. However, owing to the limitations and discomfort of traditional blood pressure (BP) detection techniques, many new cuff-less blood pressure approaches have been proposed and explored. Most of these involve arterial wave propagation theory, which is based on pulse arrival time (PAT), the time interval needed for a pulse wave to travel from the heart to some distal place on the body, such as the finger or earlobe. For this study, the Medical Information Mart for Intensive Care (MIMIC) database was used as a benchmark for PAT analysis. Many researchers who use the MIMIC database make the erroneous assumption that all the signals are synchronized. Therefore, we decided to investigate the calculation of PAT intervals in the MIMIC database and check its usefulness for evaluating BP. Our findings have important implications for the future use of the MIMIC database, especially for BP evaluation.


A fluorescent reporter for quantification and enrichment of DNA editing by APOBEC-Cas9 or cleavage by Cas9 in living cells.
Base editing is an exciting new genome engineering technology. C-to-T mutations in genomic DNA have been achieved using ribonucleoprotein complexes comprised of rat APOBEC1 single-stranded DNA deaminase, Cas9 nickase (Cas9n), uracil DNA glycosylase inhibitor (UGI), and guide (g)RNA. Here, we report the first real-time reporter system for quantification of APOBEC-mediated base editing activity in living mammalian cells. The reporter expresses eGFP constitutively as a marker for transfection or transduction, and editing restores functionality of an upstream mCherry cassette through the simultaneous processing of two gRNA binding regions that each contain an APOBEC-preferred 5'TCA target site. Using this system as both an episomal and a chromosomal editing reporter, we show that human APOBEC3A-Cas9n-UGI and APOBEC3B-Cas9n-UGI base editing complexes are more efficient than the original rat APOBEC1-Cas9n-UGI construct. We also demonstrate coincident enrichment of editing events at a heterologous chromosomal locus in reporter-edited, mCherry-positive cells. The mCherry reporter also quantifies the double-stranded DNA cleavage activity of Cas9, and may therefore be adaptable for use with many different CRISPR systems. The combination of a rapid, fluorescence-based editing reporter system and more efficient, structurally defined DNA editing enzymes broadens the versatility of the rapidly expanding toolbox of genome editing and engineering technologies.


The distribution and function of teleost IgT.
Given the uniquely close relationship between fish and aquatic environments, fish mucosal tissues are constantly exposed to a wide array of pathogenic microorganisms in the surrounding water. To maintain mucosal homeostasis, fish have evolved a distinct mucosal immune system known as mucosal-associated lymphoid tissues (MALTs). These MALTs consist of key effector cells and molecules from the adaptive immune system, such as B cells and immunoglobulins (Igs), which play crucial roles in maintaining mucosal homeostasis and defending against external pathogen infections. Until recently, three primary Ig isotypes, IgM, IgD, and IgT, have been identified in varying proportions within the mucosal secretions of teleost fish. Similar to the role of mucosal IgA in mammals and birds, teleost IgT plays a predominant role in mucosal immunity. Following the identification of the IgT gene in 2005, significant advances have been made in researching the origin, evolution, structure, and function of teleost IgT. Multiple IgT variants have been identified in various species of teleost fish, underscoring the remarkable complexity of IgT in fish. Therefore, this study provides a comprehensive review of the recent advances in various aspects of teleost IgT, including its genomic and structural features, the diverse distribution patterns within various fish mucosal tissues (the skin, gills, gut, nasal, buccal, pharyngeal, and swim bladder mucosa), its interaction with mucosal symbiotic microorganisms, and its immune responses towards diverse pathogens, including bacteria, viruses, and parasites. We also highlight the existing research gaps in the study of teleost IgT, suggesting the need for further investigation into the functional aspects of IgT and IgT+ B cells. This research is aimed at providing valuable insights into the immune functions of IgT and the mechanisms underlying the immune responses of fish against infections.


Reduced frequency of a CD14+ CD16+ monocyte subset with high Toll-like receptor 4 expression in cord blood compared to adult blood contributes to lipopolysaccharide hyporesponsiveness in newborns.
The human innate immune response to pathogens is not fully effective and mature until well into childhood, as exemplified by various responses to Toll-like receptor (TLR) agonists in newborns compared to adults. To better understand the mechanistic basis for this age-related difference in innate immunity, we compared tumor necrosis factor alpha (TNF-α) production by monocytes from cord blood (CB) and adult blood (AB) in response to LAM (lipoarabinomannan from Mycobacterium tuberculosis, a TLR2 ligand) and LPS (lipopolysaccharide from Escherichia coli, a TLR4 ligand). LPS or LAM-induced TNF-α production was 5 to 18 times higher in AB than in CB monocytes, whereas interleukin-1α (IL-1α) stimulated similar levels of TNF-α in both groups, suggesting that decreased responses to LPS or LAM in CB are unlikely to be due to differences in the MyD88-dependent signaling pathway. This impaired signaling was attributable, in part, to lower functional TLR4 expression, especially on CD14(+) CD16(+) monocytes, which are the primary cell subset for LPS-induced TNF-α production. Importantly, the frequency of CD14(+) CD16(+) monocytes in CB was 2.5-fold lower than in AB (P < 0.01). CB from Kenyan newborns sensitized to parasite antigens in utero had more CD14(+) CD16(+) monocytes (P = 0.02) and produced higher levels of TNF-α in response to LPS (P = 0.004) than CB from unsensitized Kenyan or North American newborns. Thus, a reduced CD14(+) CD16(+) activated/differentiated monocyte subset and a correspondingly lower level of functional TLR4 on monocytes contributes to the relatively low TNF-α response to LPS observed in immunologically naive newborns compared to the response in adults.


Dynamic evolution of size and colour in the highly specialized Zodarion ant-eating spiders.
Ecological specialists constitute relevant case studies for understanding the mechanisms, potential and limitations of evolution. The species-rich and strictly myrmecophagous spiders of the genus Zodarion show diversified defence mechanisms, including myrmecomorphy of different ant species and nocturnality. Through Hybridization Capture Using RAD Probes (hyRAD), a phylogenomic technique designed for sequencing poorly preserved specimens, we reconstructed a phylogeny of Zodarion using 52 (approx. a third of the nominal) species that cover its phylogenetic and distributional diversity. We then estimated the evolution of body size and colour, traits that have diversified noticeably and are linked to defence mechanisms, across the group. Our genomic matrix of 300 loci led to a well-supported phylogenetic hypothesis that uncovered two main clades inside Zodarion. Ancestral state estimation revealed the highly dynamic evolution of body size and colour across the group, with multiple transitions and convergences in both traits, which we propose is likely indicative of multiple transitions in ant specialization across the genus. Our study will allow the informed targeted selection of Zodarion taxa of special interest for research into the group's remarkable adaptations to ant specialization. It also exemplifies the utility of hyRAD for phylogenetic studies using museum material.


Persistence and storage of activity patterns in spiking recurrent cortical networks: modulation of sigmoid signals by after-hyperpolarization currents and acetylcholine.
Many cortical networks contain recurrent architectures that transform input patterns before storing them in short-term memory (STM). Theorems in the 1970's showed how feedback signal functions in rate-based recurrent on-center off-surround networks control this process. A sigmoid signal function induces a quenching threshold below which inputs are suppressed as noise and above which they are contrast-enhanced before pattern storage. This article describes how changes in feedback signaling, neuromodulation, and recurrent connectivity may alter pattern processing in recurrent on-center off-surround networks of spiking neurons. In spiking neurons, fast, medium, and slow after-hyperpolarization (AHP) currents control sigmoid signal threshold and slope. Modulation of AHP currents by acetylcholine (ACh) can change sigmoid shape and, with it, network dynamics. For example, decreasing signal function threshold and increasing slope can lengthen the persistence of a partially contrast-enhanced pattern, increase the number of active cells stored in STM, or, if connectivity is distance-dependent, cause cell activities to cluster. These results clarify how cholinergic modulation by the basal forebrain may alter the vigilance of category learning circuits, and thus their sensitivity to predictive mismatches, thereby controlling whether learned categories code concrete or abstract features, as predicted by Adaptive Resonance Theory. The analysis includes global, distance-dependent, and interneuron-mediated circuits. With an appropriate degree of recurrent excitation and inhibition, spiking networks maintain a partially contrast-enhanced pattern for 800 ms or longer after stimuli offset, then resolve to no stored pattern, or to winner-take-all (WTA) stored patterns with one or multiple winners. Strengthening inhibition prolongs a partially contrast-enhanced pattern by slowing the transition to stability, while strengthening excitation causes more winners when the network stabilizes.


Scoping review of patient- and family-oriented outcomes and measures for chronic pediatric disease.
Improvements in health care for children with chronic diseases must be informed by research that emphasizes outcomes of importance to patients and families. To support a program of research in the field of rare inborn errors of metabolism (IEM), we conducted a broad scoping review of primary studies that: (i) focused on chronic pediatric diseases similar to IEM in etiology or manifestations and in complexity of management; (ii) reported patient- and/or family-oriented outcomes; and (iii) measured these outcomes using self-administered tools. We developed a comprehensive review protocol and implemented an electronic search strategy to identify relevant citations in Medline, EMBASE, DARE and Cochrane. Two reviewers applied pre-specified criteria to titles/abstracts using a liberal accelerated approach. Articles eligible for full-text review were screened by two independent reviewers with discrepancies resolved by consensus. One researcher abstracted data on study characteristics, patient- and family-oriented outcomes, and self-administered measures. Data were validated by a second researcher. 4,118 citations were screened with 304 articles included. Across all included reports, the most-represented diseases were diabetes (35%), cerebral palsy (23%) and epilepsy (18%). We identified 43 unique patient- and family-oriented outcomes from among five emergent domains, with mental health outcomes appearing most frequently. The studies reported the use of 405 independent self-administered measures of these outcomes. Patient- and family-oriented research investigating chronic pediatric diseases emphasizes mental health and appears to be relatively well-developed in the diabetes literature. Future research can build on this foundation while identifying additional outcomes that are priorities for patients and families.


The use of a video laryngoscope by emergency medicine residents is associated with a reduction in esophageal intubations in the emergency department.
The purpose of this investigation was to compare the incidence of esophageal intubations (EIs) when emergency medicine (EM) residents used a direct laryngoscope (DL) versus a video laryngoscope (VL) for intubation attempts in the emergency department (ED). Prospectively collected continuous quality improvement data on tracheal intubations performed by EM residents in an academic ED over a 6-year period were retrospectively analyzed. Following each intubation, EM residents completed a data form with patient, intubation, and operator characteristics. Data collected included the method of intubation, drugs used, device(s) used, number of attempts, outcome of each attempt, occurrence of EIs, and occurrence of adverse events (hypoxemia, aspiration, dysrhythmia, hypotension, and cardiac arrest). The incidence of EI was compared between intubation attempts with a DL and with a VL (GlideScope(®) or C-MAC(®) ). Propensity score matching and conditional logistic regression were used to analyze the association between the intubation device (DL vs. VL) and EI. Over the 6-year period, 2,677 patients underwent 3,425 intubation attempts by EM residents with a DL or a VL. A DL was used in 1,530 attempts (44.7%) and a VL was used in 1,895 attempts (55.3%). There were 96 recognized EIs (2.8%). The incidence of EI when using a DL was 78 of 1,530 attempts (5.1%; 95% confidence interval [CI] = 4.1% to 6.3%) and when using a VL was 18 of 1,895 attempts (1.0%; 95% CI = 0.6% to 1.5%). Based on the propensity score matched analysis, the odds ratio for the occurrence of an EI for DL versus VL was 6.9 (95% CI = 3.3 to 14.4). Patients who had inadvertent EIs had a higher incidence of adverse events (49.5%; 95% CI = 38.9% to 60.0%) than patients in which EI did not occur (19.8%; 95% CI = 18.3% to 21.4%). The use of a VL by EM residents during an intubation attempt in the ED was associated with significantly fewer EIs compared to when a DL was used. Patients who had inadvertent EIs had significantly more adverse events than those who did not have EIs. EM residency training programs should consider using VLs for ED intubations to maximize patient safety when EM residents are performing intubation.


Biosupported bimetallic Pd-Au nanocatalysts for dechlorination of environmental contaminants.
Biologically produced monometallic palladium nanoparticles (bio-Pd) have been shown to catalyze the dehalogenation of environmental contaminants, but fail to efficiently catalyze the degradation of other important recalcitrant halogenated compounds. This study represents the first report of biologically produced bimetallic Pd/Au nanoparticle catalysts. The obtained catalysts were tested for the dechlorination of diclofenac and trichlorethylene. When aqueous bivalent Pd(II) and trivalent Au(III) ions were both added to concentrations of 50 mg L(-1) and reduced simultaneously by Shewanella oneidensis in the presence of H(2), the resulting cell-associated bimetallic nanoparticles (bio-Pd/Au) were able to dehalogenate 78% of the initially added diclofenac after 24 h; in comparison, no dehalogenation was observed using monometallic bio-Pd or bio-Au. Other catalyst-synthesis strategies did not show improved dehalogenation of TCE and diclofenac compared with bio-Pd. Synchrotron-based X-ray diffraction, (scanning) transmission electron microscopy and energy dispersive X-ray spectroscopy indicated that the simultaneous reduction of Pd and Au supported on cells of S. oneidensis resulted in the formation of a unique bimetallic crystalline structure. This study demonstrates that the catalytic activity and functionality of possibly environmentally more benign biosupported Pd-catalysts can be improved by coprecipitation with Au.


Anatomic, functional and molecular imaging in lung cancer precision radiation therapy: treatment response assessment and radiation therapy personalization.
This article reviews key imaging modalities for lung cancer patients treated with radiation therapy (RT) and considers their actual or potential contributions to critical decision-making. An international group of researchers with expertise in imaging in lung cancer patients treated with RT considered the relevant literature on modalities, including computed tomography (CT), magnetic resonance imaging (MRI) and positron emission tomography (PET). These perspectives were coordinated to summarize the current status of imaging in lung cancer and flag developments with future implications. Although there are no useful randomized trials of different imaging modalities in lung cancer, multiple prospective studies indicate that management decisions are frequently impacted by the use of complementary imaging modalities, leading both to more appropriate treatments and better outcomes. This is especially true of 18F-fluoro-deoxyglucose (FDG)-PET/CT which is widely accepted to be the standard imaging modality for staging of lung cancer patients, for selection for potentially curative RT and for treatment planning. PET is also more accurate than CT for predicting survival after RT. PET imaging during RT is also correlated with survival and makes response-adapted therapies possible. PET tracers other than FDG have potential for imaging important biological process in tumors, including hypoxia and proliferation. MRI has superior accuracy in soft tissue imaging and the MRI Linac is a rapidly developing technology with great potential for online monitoring and modification of treatment. The role of imaging in RT-treated lung cancer patients is evolving rapidly and will allow increasing personalization of therapy according to the biology of both the tumor and dose limiting normal tissues.


Targeting Medication Non-Adherence Behavior in Selected Autoimmune Diseases: A Systematic Approach to Digital Health Program Development.
29 autoimmune diseases, including Rheumatoid Arthritis, gout, Crohn's Disease, and Systematic Lupus Erythematosus affect 7.6-9.4% of the population. While effective therapy is available, many patients do not follow treatment or use medications as directed. Digital health and Web 2.0 interventions have demonstrated much promise in increasing medication and treatment adherence, but to date many Internet tools have proven disappointing. In fact, most digital interventions continue to suffer from high attrition in patient populations, are burdensome for healthcare professionals, and have relatively short life spans. Digital health tools have traditionally centered on the transformation of existing interventions (such as diaries, trackers, stage-based or cognitive behavioral therapy programs, coupons, or symptom checklists) to electronic format. Advanced digital interventions have also incorporated attributes of Web 2.0 such as social networking, text messaging, and the use of video. Despite these efforts, there has not been little measurable impact in non-adherence for illnesses that require medical interventions, and research must look to other strategies or development methodologies. As a first step in investigating the feasibility of developing such a tool, the objective of the current study is to systematically rate factors of non-adherence that have been reported in past research studies. Grounded Theory, recognized as a rigorous method that facilitates the emergence of new themes through systematic analysis, data collection and coding, was used to analyze quantitative, qualitative and mixed method studies addressing the following autoimmune diseases: Rheumatoid Arthritis, gout, Crohn's Disease, Systematic Lupus Erythematosus, and inflammatory bowel disease. Studies were only included if they contained primary data addressing the relationship with non-adherence. Out of the 27 studies, four non-modifiable and 11 modifiable risk factors were discovered. Over one third of articles identified the following risk factors as common contributors to medication non-adherence (percent of studies reporting): patients not understanding treatment (44%), side effects (41%), age (37%), dose regimen (33%), and perceived medication ineffectiveness (33%). An unanticipated finding that emerged was the need for risk stratification tools (81%) with patient-centric approaches (67%). This study systematically identifies and categorizes medication non-adherence risk factors in select autoimmune diseases. Findings indicate that patients understanding of their disease and the role of medication are paramount. An unexpected finding was that the majority of research articles called for the creation of tailored, patient-centric interventions that dispel personal misconceptions about disease, pharmacotherapy, and how the body responds to treatment. To our knowledge, these interventions do not yet exist in digital format. Rather than adopting a systems level approach, digital health programs should focus on cohorts with heterogeneous needs, and develop tailored interventions based on individual non-adherence patterns.


Production of a fluorescence resonance energy transfer (FRET) biosensor membrane for microRNA detection.
MicroRNAs (miRNAs) play a key role in regulating gene expression but can be associated with abnormalities linked to carcinogenesis and tumor progression. Hence there is increasing interest in developing methods to detect these non-coding RNA molecules in the human circulation system. Here, a novel FRET miRNA-195 targeting biosensor, based on silica nanofibers incorporated with rare earth-doped calcium fluoride particles (CaF2:Yb,Ho@SiO2) and gold nanoparticles (AuNPs), is reported. The formation of a sandwich structure, as a result of co-hybridization of the target miRNA which is captured by oligonucleotides conjugated at the surface of CaF2:Yb,Ho@SiO2 fibers and AuNPs, brings the nanofibers and AuNPs in close proximity and triggers the FRET effect. The intensity ratio of green to red emission, I541/I650, was found to decrease linearly upon increasing the concentration of the target miRNA and this can be utilized as a standard curve for quantitative determination of miRNA concentration. This assay offers a simple and convenient method for miRNA quantification, with the potential for rapid and early clinical diagnosis of diseases such as breast cancer.


The interaction between primate blood and mouse islets induces accelerated clotting with islet destruction.
Mouse islets transplanted under the renal subcapsular space of cynomolgus monkeys are subject to a form of hyperacute rejection, the mechanism of which is unclear. As islets are in contact with whole blood at the time of transplantation, the effect of platelets and the coagulation cascade on islet destruction was assessed. Coagulation was assessed using thromboelastography on citrated/recalcified human blood samples with freshly isolated C57/Bl6 mouse islets. A dynamic islet perifusion system was used to assess the effect of islets on blood cells and coagulation factors. Cytotoxicity was evaluated using (51)Cr labelled islets incubated with human blood and islet destruction was also evaluated using a histological grading system. Continuous PO(2) measurements were made in a static incubation system to assess the role of hypoxia in islet destruction. Mouse islets incubated in human blood induced accelerated coagulation and rapid consumption of platelets within 15 min. Within 1 h of incubation, 52% of mouse islets exposed to xenogeneic human blood showed features of severe damage with necrosis when compared with islets incubated in syngeneic blood. Specific lysis of the xenogeneic islets was demonstrable (Mean percentage lysis: 48%, P < 0.05 vs. control) after 4 h incubation in human blood. Oxygen levels remained constant at a level adequate to maintain islet viability in separate experiments. Mouse islets induce rapid activation of the clotting cascade and platelet consumption in vitro when exposed to human blood, which correlated with histological evidence of significant destruction demonstrable within 1 h of exposure to human or non-human primate blood. This in vitro model has features which appear to correlate with the islet destruction seen in vivo and could be a useful model for the study of the mechanisms underlying the rapid destruction of xenogeneic islets in primate recipients.


Common and uncommon features of focal splenic lesions on contrast-enhanced ultrasound: a pictorial review.
The characterization of focal splenic lesions by ultrasound can be quite challenging. The recent introduction of contrast-enhanced ultrasound (CEUS) has come to play a valuable role in the field of imaging splenic pathologies, offering the possibility of an ionizing radiation-free investigation. Because CEUS has been incorporated into everyday clinical practice, malignant diseases such as focal lymphomatous infiltration, metastatic deposits, benign cysts, traumatic fractures, and hemangiomas can now be accurately depicted and characterized without the need for further imaging. More specifically, splenic traumatic fractures do not require additional imaging by computed tomography (with ionizing radiation exposure) for follow-up, because splenic fractures and their complications are safely imaged with CEUS. In the new era of CEUS, more patients benefit from radiation-free investigation of splenic pathologies with high diagnostic accuracy.


Operational effectiveness of single-dose nevirapine in preventing mother-to-child transmission of HIV.
To determine the operational effectiveness of the South African programme for preventing mother-to-child transmission (PMTCT) of HIV in reducing rates of early transmission of infection. Participants were mother-infant pairs who participated in the South African PMTCT programme between October 2002 and November 2004. This was a prospective cohort study. Three sites in different provinces were selected to represent differences in socioeconomic status and HIV prevalence. Data on antenatal care and labour ward care were obtained from maternal interviews and from reviews of medical records. A total of 665 mother-infant pairs in which the mother was HIV-positive were recruited and 588 (88.4%) were followed up at 3 or 4 weeks postpartum to determine the HIV status and vital status of the infant. Rural participants were significantly poorer and their health care was significantly worse. Women of higher socioeconomic status and those who received better counselling were more likely to be treated with nevirapine. Rates of early HIV transmission ranged from 8.6% to 13.7%. Maternal viral load was the only statistically significant risk factor for transmission. After adjusting for maternal viral load and prevalence of low birth weight, the odds of transmission were 1.8 times higher at the rural site. Controlling for having had > or = 4 antenatal visits and any delivery complication reduced the odds of transmission to 1.5 higher at the rural site. Rates of early transmission of HIV in an operational setting using single-dose nevirapine administered both to mother and child are similar to those obtained in clinical trials. Scaling up access to antiretroviral regimens for women will further reduce transmission to infants.


Insertion torque and resonance frequency analysis of dental implant systems in an animal model with loaded implants.
The aim of this study was to compare insertion torque and resonance frequency analysis of different implant systems in an animal model with loaded implants. Three types of Brånemark implants (machined MkIII, TiUnite MkIII, and MkIV) and 2 types of Straumann implants (sandblasted, large-grit, acid-etched [SLA] and titanium plasma-sprayed [TPS]) were studied. Thirty-two implants of each type (n = 160) were placed in 16 beagle dogs. Maximum insertion torque values were recorded. After a healing period of 8 weeks, the implants were loaded for 3 months; the animals were then sacrificed. At placement, after healing, and at the end of the loading phase, resonance frequency analysis was performed and implant stability quotients (ISQs) were recorded. Higher insertion torque values were seen for the conical MkIV than for the MkIII. No difference was seen between the Brånemark and Straumann implants on the basis of ISQ value at placement. ISQ and insertion torque values were lower for the cylindric Straumann implants than for the self-tapping implants. For all implant systems a significant decrease in median ISQ was observed, with a median decrease ranging from 3 to 6. ISQ values for self-tapping implants remained stable after loading, whereas the ISQ values for non-self-tapping cylinders decreased. The maximum insertion torque values for failed and successful implants were not significantly different. Significantly higher ISQ values at placement were seen for successful implants (P =.003). Based on this model for ISQ, a threshold of 65.5 was identified, with a sensitivity of 83% and specificity of 61% for prediction of implant loss. ISQ values at the start of loading were not predictive of implant loss in the loading period. Caution should be used when judging implant systems on the basis of resonance frequency analysis and torque measurement.


Diagnosis of peripheral lung cancer with three echoic features via endobronchial ultrasound.
Endobronchial ultrasonography (EBUS) is useful in localizing peripheral lung lesions. Previous reports have revealed that several characteristic echoic patterns correlate well with the histopathologic findings of benign and malignant lesions. Therefore, EBUS may be also useful in the differential diagnosis of malignant lesions of the lung. To assess the feasibility of EBUS in the differential diagnosis between malignant and benign lesions by the following three characteristic echoic features indicating malignancy: continuous margin; absence of a linear-discrete air bronchogram; and heterogeneous echogenicity. EBUS images from 224 patients who undergone bronchoscopy for a peripheral lung lesion were analyzed. The sensitivity and specificity for each echoic feature or in combination in diagnosing malignancy or benignity were determined. Continuous margin, absence of linear-discrete air bronchogram, and heterogeneous echogenicity are three echoic features indicating malignancy. The absence of linear-discrete air bronchogram has the highest sensitivity in the diagnosis of malignancy (91.9%), but the lowest specificity (62.4%). By contrast, a well-defined margin has the highest specificity (93.1%), but the lowest sensitivity (27.6%). The sensitivity and specificity for heterogeneous echogenicity are intermediate (65.0% and 90.1%, respectively). The negative predictive value for the malignancy of a lesion with none of these three echoic features is 93.7%. The positive predictive value for malignancy of a lesion with any two of the three echoic features is 89.2%. These results indicate that EBUS is useful as an adjunct in lung cancer diagnosis, especially when peripheral lung lesions are not visible in traditional bronchoscopy.


Cyproheptadine use in hepatocellular carcinoma.
This study was conducted to compare the effectiveness of Cyproheptadine (CY) use in patients with different stages of HCC who received different therapeutic modalities; such a comparison has not been conducted by previous large, prospective, randomized studies. We conducted a cohort study using the Taiwan Cancer Registry Database for analysis. We included patients diagnosed as having HCC from January 1, 2002, to December 31, 2011. The patient cohort comprised those who received different treatments, and we compared patients who received CY with those who did not. In total, 70,885 patients were included, and the mean follow-up duration was 1.95 years. The adjusted hazard ratio (aHR) of all-cause deaths significantly decreased in all stages in the patients who received palliative treatments with CY use compared with those who received palliative treatments without CY use (all P < 0.0001 and aHR = 0.76, 0.80, 0.66, and 0.66 for stages I, II, III, and IV, respectively). Among the patients who received no treatment, CY use alone reduced the risk of all-cause deaths in stages I-IV (all P < 0.0001 and aHR = 0.61, 0.57, 0.54, and 0.52 for stages I, II, III, and IV, respectively). Among the patients with clinical stage I-II HCC (as determined by the American Joint Committee on Cancer) who received curative treatments, CY use did not reduce all-cause deaths. CY use might improve survival in patients with HCC receiving palliative treatments or no treatment regardless of clinical stages.


Surface dangling bonds are a cause of B-type blinking in Si nanoparticles.
Exponential blinking statistics was reported in oxidized Si nanoparticles and the switching mechanism was attributed to the activation and deactivation of unidentified nonradiative recombination centers. Using ab initio calculations we predicted that Si dangling bonds at the surface of oxidized nanoparticles introduce defect states which, depending on their charge and local stress conditions, may give rise to ON and OFF states responsible for exponential blinking statistics. Our results are based on first principles calculations of charge transition levels, single particle energies, and radiative and nonradiative lifetimes of dangling bond defects at the surface of oxidized silicon nanoparticles under stress.


Portal vein thrombosis in 33 dogs: 1998-2011.
Portal vein thrombosis (PVT) has been reported infrequently in dogs. To characterize the presentation, associated disease conditions, and outcome in dogs with PVT. Client-owned dogs with a diagnosis of PVT and a complete medical record. Records were retrospectively analyzed for presentation, history, physical examination, clinicopathologic data, diagnostic imaging, treatment, and outcome. Thirty-three dogs were included. Common clinical signs were vomiting, diarrhea, abdominal pain, ascites, and signs of hypovolemic shock. Associated disease conditions included hepatic (14/33), neoplastic (7/33), immune (5/33), and infectious (4/33) diseases, protein-losing nephropathy (3/33), hyperadrenocorticism (2/33), protein-losing enteropathy (1/33), and pancreatitis (1/33). Fourteen dogs were receiving glucocorticoids at the time of diagnosis. Twenty-nine dogs had at least 1 predisposing condition for venous thrombosis, and 11 had 2 or more. Thrombocytopenia (24/33), increased liver enzyme activity (23/33), and hypoalbuminemia (20/33) were common laboratory abnormalities. Clinical syndromes at the time of PVT diagnosis included shock (16/33), systemic inflammatory response syndrome (SIRS), (13/33) and disseminated intravascular coagulation (3/33). Twenty-four dogs had acute and 9 had chronic PVT. Multiple thrombi were found in 17/33 dogs. Nineteen dogs survived to discharge. Dogs treated with anticoagulant therapy were more likely, whereas those with acute PVT, multiple thromboses or SIRS were less likely to survive. Hepatic disease is a common pre-existing condition in dogs with PVT. PVT should be considered in dogs with risk factors for venous thrombosis presenting with abdominal pain, ascites, and thrombocytopenia. Studies evaluating anticoagulant therapy in the management of PVT are warranted.


Crystal structure of an arginase-like protein from Trypanosoma brucei that evolved without a binuclear manganese cluster.
The X-ray crystal structure of an arginase-like protein from the parasitic protozoan Trypanosoma brucei, designated TbARG, is reported at 1.80 and 2.38 Å resolution in its reduced and oxidized forms, respectively. The oxidized form of TbARG is a disulfide-linked hexamer that retains the overall architecture of a dimer of trimers in the reduced form. Intriguingly, TbARG does not contain metal ions in its putative active site, and amino acid sequence comparisons indicate that all but one of the residues required for coordination to the catalytically obligatory binuclear manganese cluster in other arginases are substituted here with residues incapable of metal ion coordination. Therefore, the structure of TbARG is the first of a member of the arginase/deacetylase superfamily that is not a metalloprotein. Although we show that metal binding activity is easily reconstituted in TbARG by site-directed mutagenesis and confirmed in X-ray crystal structures, it is curious that this protein and its parasitic orthologues evolved away from metal binding function. Knockout of the TbARG gene from the genome demonstrated that its function is not essential to cultured bloodstream-form T. brucei, and metabolomics analysis confirmed that the enzyme has no role in the conversion of l-arginine to l-ornithine in these cells. While the molecular function of TbARG remains enigmatic, the fact that the T. brucei genome encodes only this protein and not a functional arginase indicates that the parasite must import l-ornithine from its host to provide a source of substrate for ornithine decarboxylase in the polyamine biosynthetic pathway, an active target for the development of antiparasitic drugs.


Microorganisms in closed periapical lesions.
The purpose of this study was to investigate the microorganisms of strictly selected closed periapical lesions associated with both refractory endodontic therapy and pulpal calcification. Definitive criteria were established that assured complete clinical isolation of the periapical lesion from the oral and periodontal environment. A total of 13 criteria-referenced lesions were selected from 70 patients with endodontic surgical indications. A well controlled culturing method was used in all cases and samples were taken by one clinician at three separate sites during each surgery. Samples taken at the surgical window and within the body of the lesion served as controls, whilst a third sample was taken at the apex. In all 13 cases, samples taken from the apex yielded microorganisms comprising 63.6% obligate anaerobes and 36.4% facultative anaerobes. Prevalence of the isolated species was 31.8% for Actinomyces sp., 22.7% Propionibacterium sp., 18.2% Streptococcus sp., 13.6% Staphlyococcus sp., 4.6% Porphyromonas gingivalis, 4.6% Peptostreptococcus micros and 4.6% Gram-negative enterics. The results of this investigation indicate that closed periapical lesions associated with calcified teeth or those resistant to root canal treatment harbour bacteria. The inability to eradicate all root canal microorganisms during root canal treatment, along with anatomical factors, may allow further bacterial colonization of the root apex and surrounding periapical tissues, and consequently prevent healing.


Effect of various dentin disinfection protocols on the bond strength of resin modified glass ionomer restorative material.
Disinfection of dentin surface prior to any restorative therapy is important for the longevity of the treatment rendered. However, these dentin disinfection methods should itself not interfere with the adhesion of the restorative material. Therefore the aim of this study was to determine the effect of various dentin disinfection protocols on the shear bond strength (SBS) of resin modified glass ionomer cement (RMGIC). The occlusal surface of 40 extracted premolars were trimmed to obtain a flat dentinal surface and was randomly divided into four groups. CTRL was the control group; NaOCl was 1% sodium hypochlorite disinfection group; CHX was 2% chlorhexidine disinfection group; and PAD was the photo-activated disinfection group. Then a predetermined dimension of RMGIC was bonded to the pre-treated dentin surfaces. Following this, each sample was tested for SBS using universal testing machine at a crosshead speed of 0.5mm/min. Among the test groups, CHX showed the least reduction in SBS and NaOCl the highest reduction in SBS as compared to the control group. PAD on the other hand showed significantly lower SBS than CTRL and CHX groups, but the values were higher than the NaOCl group. Thus, it could be concluded from the present study that use of chlorhexidine based dentin disinfection does interfere with the adhesion of RMGIC. However, photo-activated disinfection should be done with caution. Moreover, sodium hypochlorite based disinfectants should be avoided prior to the use of RMGIC. Key words:Chlorhexidine, Dentin disinfection, Photo-activated disinfection, Resin modified glass ionomer cement, Shear bond strength, Sodium hypochlorite.


An improved detection method for the Rhopalosiphum padi virus (RhPV) allows monitoring of its presence in aphids and movement within plants.
Rhopalosiphum padi virus (RhPV) is an insect RNA virus that infects aphids, reducing their lifespan and fecundity. It can be transmitted vertically between aphids and horizontally via the plant. An improved detection method for the virus in aphids and plants using RT-PCR was developed; this allowed individual aphids to be tested for RhPV. Testing of R. padi aphids collected from different sites in Sweden revealed the presence of RhPV in wild aphid populations for the first time in Europe. Virus could be detected in several life stages of R. padi, including sexual individuals and eggs, establishing an over-wintering route for the virus. Using RT-PCR, systemic transport of the virus in plants was tracked. Virus spread from the aphid feeding site to all parts of the plant, including roots, within 7 days, and could be acquired by virus-free aphids feeding on the same plant.


Learning combinatorial transcriptional dynamics from gene expression data.
mRNA transcriptional dynamics is governed by a complex network of transcription factor (TF) proteins. Experimental and theoretical analysis of this process is hindered by the fact that measurements of TF activity in vivo is very challenging. Current models that jointly infer TF activities and model parameters rely on either of the two main simplifying assumptions: either the dynamics is simplified (e.g. assuming quasi-steady state) or the interactions between TFs are ignored, resulting in models accounting for a single TF. We present a novel approach to reverse engineer the dynamics of multiple TFs jointly regulating the expression of a set of genes. The model relies on a continuous time, differential equation description of transcriptional dynamics where TFs are treated as latent on/off variables and are modelled using a switching stochastic process (telegraph process). The model can not only incorporate both activation and repression, but allows any non-trivial interaction between TFs, including AND and OR gates. By using a factorization assumption within a variational Bayesian treatment we formulate a framework that can reconstruct both the activity profiles of the TFs and the type of regulation from time series gene expression data. We demonstrate the identifiability of the model on a simple but non-trivial synthetic example, and then use it to formulate non-trivial predictions about transcriptional control during yeast metabolism. http://homepages.inf.ed.ac.uk/gsanguin/ email@example.com Supplementary data are available at Bioinformatics online.


Brain natriuretic peptide and left ventricular dysfunction in chagasic cardiomyopathy.
Global left ventricular (LV) systolic dysfunction is the strongest predictor of morbidity and mortality in Chagas disease. Echocardiography is considered the gold standard for the detection of LV dysfunction, but not always available in endemic areas where chagasic cardiomyopathy is most common. Brain natriuretic peptide (BNP) is a neurohormone that has been recently described as a simple and inexpensive diagnostic and prognostic marker for patients with congestive heart failure. Chagasic patients (n = 63) and non-infected healthy individuals (n = 18) were recruited prospectively and underwent complete clinical examination, echocardiography and 24-h Holter monitoring. BNP was measured from thawed plasma samples using the Triage BNP test. We observed high levels of BNP in association with depression of LV ejection fraction, with increase of LV end-diastolic diameter and with LV premature complexes. An elevated concentration of BNP, defined as a concentration of 60 pg/ml or more, had a sensitivity of 91.7%, specificity of 82.8%, positive predictive value of 52.4%, and negative predictive value of 98% for detecting LV dysfunction (LV ejection fraction < 40%).BNP measurement using a simple, relatively inexpensive and rapid test has a promising role in identifying LV dysfunction associated with chagasic cardiomyopathy. Equally important, patients with Trypanosoma cruzi infection who have low levels of BNP level in plasma have a very low likelihood of severe cardiac involvement, and echocardiography is probably not necessary.


Weighted graph cuts without eigenvectors a multilevel approach.
A variety of clustering algorithms have recently been proposed to handle data that is not linearly separable; spectral clustering and kernel k-means are two of the main methods. In this paper, we discuss an equivalence between the objective functions used in these seemingly different methods--in particular, a general weighted kernel k-means objective is mathematically equivalent to a weighted graph clustering objective. We exploit this equivalence to develop a fast, high-quality multilevel algorithm that directly optimizes various weighted graph clustering objectives, such as the popular ratio cut, normalized cut, and ratio association criteria. This eliminates the need for any eigenvector computation for graph clustering problems, which can be prohibitive for very large graphs. Previous multilevel graph partitioning methods, such as Metis, have suffered from the restriction of equal-sized clusters; our multilevel algorithm removes this restriction by using kernel k-means to optimize weighted graph cuts. Experimental results show that our multilevel algorithm outperforms a state-of-the-art spectral clustering algorithm in terms of speed, memory usage, and quality. We demonstrate that our algorithm is applicable to large-scale clustering tasks such as image segmentation, social network analysis and gene network analysis.


Gender, Physical Self-Perception and Overall Physical Fitness in Secondary School Students: A Multiple Mediation Model.
Background: Physical self-perception is often related with better physical fitness perception in adolescents. Moreover, it is an important social cognitive perspective to provide suitable mental health in this population. However, this relationship is unequal between boys and girls. The physical fitness is a marker of health in young population. The aims of the present study were the following: (1) to compare physical self-perception and self-reported overall physical fitness (OPF) between boys and girls (gender) and body mass index (BMI) status, and (2) to determine the mediating role of all physical self-perception subscales (except physical condition) and BMI status in the link between gender and OPF in adolescent students. Methods: This cross-sectional study consisted of 85 adolescent students of secondary school between 12 and 17 years of age; 41 were boys (Mage = 14.6, SD = 1.7) and 44 were girls (Mage = 14.4, SD = 1.6). Adolescent participants completed all clinical characteristics by body composition measures (age, body weight, body height, and BMI). Physical self-perception was assessed by the physical self-perception profile (PSPP) whereas the international fitness scale (IFIS) was used to predict the self-reported OPF of adolescents in the present study. Results: Gender (boys and girls) differed significantly in all PSPP subscales and OPF, whereas the BMI status (underweight = 19 students, normal weight = 53 students, overweight/obese = 13 students) showed significant differences in all clinical characteristics, physical condition (PSPP), and OPF. A multiple mediation analysis was performed using bias corrected bootstrap. This multiple mediation analysis revealed that all PSPP subscales were significant mediators between gender and OPF: attractive body (p = 0.013), sport competence (p = 0.009), physical strength (p = 0.002), and self-confidence (p = 0.002). The total direct effect of gender on OPF was significant (p = 0.002). Moreover, the multiple mediation estimated a completely standardized indirect of X on Y for attractive body (effect = 0.109), sport competence (effect = 0.066), physical strength (effect = 0.130), and self-confidence (effect = 0.193). Conclusions: These findings contribute to understanding the link between gender and OPF in adolescent students and the mediation of physical self-perception and OPF in this relationship. In addition, strategies focused to improve self-confidence and physical self-perception are necessary in female adolescent students, because boys showed better physical self-perception in all PSPP subscales. Girls are a risk group because they report low physical self-confidence with their respective insecurity feelings and psychological disorders. Thus, personal physical self-perception must be considered as an important social cognitive perspective to provide suitable mental health in children and adolescents.


Correlation between symptom fatigue and muscular fatigue in multiple sclerosis.
The symptom of fatigue is a frequent complaint in multiple sclerosis (MS) patients. Signs of fatigability have been documented in these patients as well. However, correlation with signs of objective fatigue had not been clarified in MS. The aim of this study was to ascertain the existence of muscular fatigue in multiple sclerosis patients, and to find out if there is a correlation between the subjective symptom of fatigue and muscular fatigue. Fifty MS patients and 50 age and sex matched volunteers were studied using isometric and isotonic tests using the dominant hand. Strength was studied in the baseline condition and also after recovery of either an isotonic (experiment A) or isometric effort (experiment B). Maximum strength, strength in relationship to weight, slope of fatigability in 11 consecutive contractions, and strength and duration of a maximum effort were calculated. Fatigue as a symptom was measured using the Fatigue Severity Scale (FSS) and the Fatigue Descriptive Scale (FDS). Non-parametric techniques were used for the statistical analysis. Patients with MS had less isometric and isotonic strength, but the recovery was the same as recovery in the control group. There was a negative linear correlation between the symptom of fatigue and the baseline strength. In conclusion, this study supports the existence of signs of muscular fatigue in MS patients. However, the recovery after exercise is normal. The correlation between the baseline scores in strength and the symptom of fatigue suggest that the same cause (probably pyramidal deficits) may be involved in both of them. Copyright 1998 Lippincott Williams & Wilkins


Psychosocial Hazard Analysis in a Heterogeneous Workforce: Determinants of Work Stress in Blue- and White-Collar Workers of the European Steel Industry.
The European steel industry's workforce is highly heterogeneous and consists of various occupational groups, presumably facing different psychosocial stressors. The few existing studies on the subject mainly focused on physical constraints of blue-collar workers, whereas the supposable psychosocial workload received only little research attention. This is remarkable considering the challenges associated with statutory required risk assessment of psychosocial hazards. Valid measures of hazard analysis must account for various stressors and reliably identify them, also between occupational groups. The present study, based on a sample of blue- and white-collar workers (N = 124) from the European steel industry, aims to provide a first insight into psychosocial stressors and strain at work in this rarely researched industrial sector. Furthermore, two well-known theoretical roadmaps in job analysis are examined regarding their utility for risk assessment in heterogeneous workforces: the German standard version of the Copenhagen Psychosocial Questionnaire (COPSOQ) and the short version of the effort-reward imbalance questionnaire. Hierarchical multiple regression analyses revealed that the COPSOQ was better suited to predict various strain indices in the present sample. Especially stressors relating to socioemotional aspects, such as work-privacy conflict, revealed a reasonable impact, indicating the need for comprehensive solutions at the organizational level instead of solutions focusing on single workplaces. To conclude, a broadly diversified and validated approach in psychosocial risk assessment is needed to adequately assess the variety of psychosocial factors at work and in different occupational groups.


Molecular Characterization of Cotrimoxazole Resistance Genes and Their Associated Integrons in Clinical Isolates of Gram-Negative Bacteria from Tanzania.
Cotrimoxazole is widely used, particularly as a prophylactic drug in HIV patients. We assessed resistance mechanisms among cotrimoxazole resistant-Gram negative bacterial isolates (n = 123) obtained from blood (n = 69) and urine (n = 54) from Tanzanian patients. sul genes were detected in 98% (121/123) of the isolates. Coexistence of sul1 and sul2 was common (49/123). The dfr genes were found in 63% (77/123) of all isolates. sul1, dfrA15, and dfrA5 genes predominated among Klebsiella pneumoniae, while sul2 and dfrA1 genes were frequent in Escherichia coli isolates. Two isolates, both K. pneumoniae, carried sul3. Integrons were detected in 81.3% (100/123) of all isolates. Class 1 integrons were found in 95% (42/44), 53% (23/43), and 80.6% (25/31) of K. pneumoniae, E. coli, and other Enterobacteriaceae isolates, respectively. Class 2 integrons were found in 14% of E. coli, but not in K. pneumoniae. All sul1 genes in K. pneumoniae were carried in class 1 integrons. Gene cassette arrays dfrA5 and dfrA15-aadA1 were most frequently associated with class 1 integrons, while class 2 integrons contained only dfrA1-sat2-aadA1 gene cassettes. This is the first report of sul3 gene in K. pneumoniae from human sources. The finding that mechanisms differ between E. coli and K. pneumoniae may broaden our understanding of cotrimoxazole resistance.


Dynamically produced refractive-index variations with thickness of volume holograms in electrooptic crystals.
Dynamic theory of volume holography is used to calculate the variations in the thickness direction of the hologram-constituting refractive-index modulation and the externally observable effects of these variations in electrooptic materials. It is shown that thick holographic gratings may exhibit significant amplitude and grating phase variations with thickness including amplitude sign reversal. These nonuniformities strongly affect holographic grating recording and readout characteristics such as maximum possible diffraction efficiency and angular selectivity. Thus a variety of grating applications will be affected by these nonuniformities.


Sequential anisotropic multichannel Wiener filtering with Rician bias correction applied to 3D regularization of DWI data.
It has been shown that the tensor calculation is very sensitive to the presence of noise in the acquired images, yielding to very low quality Diffusion Tensor Images (DTI) data. Recent investigations have shown that the noise present in the Diffusion Weighted Images (DWI) causes bias effects on the DTI data which cannot be corrected if the noise characteristic is not taken into account. One possible solution is to increase the minimum number of acquired measurements (which is 7) to several tens (or even several hundreds). This has the disadvantage of increasing the acquisition time by one (or two) orders of magnitude, making the process inconvenient for a clinical setting. We here proposed a turn-around procedure for which the number of acquisitions is maintained but, the DWI data are filtered prior to determining the DTI. We show a significant reduction on the DTI bias by means of a simple and fast procedure which is based on linear filtering; well-known drawbacks of such filters are circumvented by means of anisotropic neighborhoods and sequential application of the filter itself. Information of the first order probability density function of the raw data, namely, the Rice distribution, is also included. Results are shown both for synthetic and real datasets. Some error measurements are determined in the synthetic experiments, showing how the proposed scheme is able to reduce them. It is worth noting a 50% increase in the linear component for real DTI data, meaning that the bias in the DTI is considerably reduced. A novel fiber smoothness measure is defined to evaluate the resulting tractography for real DWI data. Our findings show that after filtering, fibers are considerably smoother on the average. Execution times are very low as compared to other reported approaches which allows for a real-time implementation.


Mutations in motB suppressible by changes in stator or rotor components of the bacterial flagellar motor.
Five proteins (MotA, MotB, FliG, FliM and FliN) may be involved in energizing flagellar rotation in Escherichia coli. To study interactions between the Mot proteins, and between them and the three Fli proteins of the switch-motor complex, we have isolated extragenic suppressors of dominant and partially dominant motB missense mutations. Four of the 13 motB mutations yielded partially allele-specific suppressors. Of the suppressing mutations, 57 are in the motA gene, eight are in fliG, and one is in fliM; no suppressor was identified in fliN. The prevalence of suppressors in fliG suggests that FliG interacts rather directly with the Mot proteins. The behaviour of cells in tethering and swarm assays indicates that the motA suppressors are more efficient than the fliG or fliM suppressors. Some of the suppressing mutations themselves confer distinctive phenotypes in motB+ cells. We propose a model in which mutations affecting residues in or near the putative peptidoglucan-binding region of MotB misalign the stator relative to the rotor. We suggest that most of the suppressors restore motility by introducing compensatory realignments in MotA or FliG.


Fascin expression in colorectal carcinomas.
The purpose of this study was to investigate the significance of fascin expression in colorectal carcinoma. This is a retrospective study of 167 consecutive, well-documented cases of primary colorectal adenocarcinoma for which archival material of surgical specimens from primary tumor resections were available. We chose a representative tissue sample block and examined fascin expression by immunohistochemistry using a primary antibody against "fascin". We calculated the "immunohistochemical score (IHS)" of fascin for each case, which was calculated from the multiplication of scores for the percentage of stained cells and the staining intensity. Fascin immunoreactivity was observed in 59 (35.3%) of all cases with strong reactivity in 24 (14.4%), moderate reactivity in 25 (14.9%) and weak reactivity in 10 (6.0%) cases. Strong/moderate immunoreactivities were mostly observed in invasive fronts of the tumors or in both invasive and other areas. Fascin immunoreactivity scores were significantly higher in tumors with lymph node metastasis (p:0.002) and advanced stage presentation (p:0.007). There was no relation between fascin expression and age, gender, depth of invasion, distant metastasis or histological grade (p>0.05). There was a higher and statistically significant correlation between fascin immunoreactivity in the invasive borders of tumors and lymph node metastasis (r:0.747, p:0.005). In stage III/IV tumors, two-year survival was 92.2% in tumors without fascin immunoreactivity, and only 60.0% in tumors with a fascin IHS>10 (p:0.003). These findings suggest that fascin is heterogeneously expressed in approximately one third of colorectal carcinomas with a significant association with lymph node metastasis, tumor stage and location. Moreover, these results indicate that fascin may have a role in the lymph node metastasis of colorectal carcinomas.


Pathogenic 12-kb copy-neutral inversion in syndromic intellectual disability identified by high-fidelity long-read sequencing.
We report monozygotic twin girls with syndromic intellectual disability who underwent exome sequencing but with negative pathogenic variants. To search for variants that are unrecognized by exome sequencing, high-fidelity long-read genome sequencing (HiFi LR-GS) was applied. A 12-kb copy-neutral inversion was precisely identified by HiFi LR-GS after trio-based variant filtering. This inversion directly disrupted two genes, CPNE9 and BRPF1, the latter of which attracted our attention because pathogenic BRPF1 variants have been identified in autosomal dominant intellectual developmental disorder with dysmorphic facies and ptosis (IDDDFP), which later turned out to be clinically found in the twins. Trio-based HiFi LR-GS together with haplotype phasing revealed that the 12-kb inversion occurred de novo on the maternally transmitted chromosome. This study clearly indicates that submicroscopic copy-neutral inversions are important but often uncharacterized culprits in monogenic disorders and that long-read sequencing is highly advantageous for detecting such inversions involved in genetic diseases.


Calf Compartment Syndrome associated with the Use of an Intra-osseous Line in an Adult Patient: A Case Report.
We present a case of a lower limb compartment syndrome associated with the use of an intra-osseous line inserted into the proximal tibia in an adult patient. An unconscious 59-year old male with multiple injuries presented to our Emergency Department after a road traffic accident. Bilateral proximal tibial intra osseous-lines were inserted due to poor venous access. After resuscitation his left leg was noted to be tense and swollen with absent pulses. Acute compartment syndrome was diagnosed both clinically and with compartment pressure measurement. Two incision fasciotomy on his left lower leg was performed. Intra osseous-lines in the proximal tibia are increasingly used in adult patients in the pre-hospital setting by paramedics and emergency physicians. Their use, along with the possible complications of these devices, such as the development of compartment syndrome or osteomyelitis leading to amputation, is well reported in the paediatric literature. To the best of our knowledge, there have not been any previous reports of complications in the adult patient. We present a case of lower leg compartment syndrome developing from the use of an intra-osseous line in the proximal tibia in an adult patient. With the increasing use of intra-osseous lines in adult patients, clinicians should be aware of the possibility of developing compartment syndrome which may lead to disability or amputation in severe cases.


Ketoconazole blocks testosterone synthesis.
Ketoconazole, a new oral drug used to treat systemic and superficial mycoses, inhibits sterol synthesis in fungi. The development of gynecomastia in two patients prompted us to investigate the effect of the drug on testosterone production. After a 200-, 400-, or 600-mg dose, volunteer male testosterone serum concentrations fell markedly, but returned toward baseline eight to 24 hours later as ketoconazole serum concentrations waned. A marked but transient drop in testosterone levels occurred in patients receiving long-term therapy, and continuous testosterone depression was noted in one. A block of synthesis was demonstrated in vitro. Ketoconazole at concentrations achievable in serum with currently used doses blocked basal and gonadotropin-stimulated testosterone production by rat Leydig cells. The diminution of testosterone synthesis could be significant as further therapeutic trials may use larger doses or more than once-daily administration. The paucity of reports of endocrinologic toxicity may relate to the "escape" from the block demonstrated in vivo.


Physiotherapy in intensive care: an updated systematic review.
Although physiotherapy is frequently provided to patients in the ICU, its role has been questioned. The purpose of this systematic literature review, an update of one published in 2000, was to examine the evidence concerning the effectiveness of physiotherapy for adult, intubated patients who are mechanically ventilated in the ICU. The main literature search was undertaken on PubMed, with secondary searches of MEDLINE, CINAHL, Embase, the Cochrane Library, and the Physiotherapy Evidence Database. Only papers published from 1999 were included. No limitations were placed on study design, intervention type, or outcomes of clinical studies; nonsystematic reviews were excluded. Items were checked for relevance and data extracted from included studies. Marked heterogeneity of design precluded statistical pooling of results and led to a descriptive review. Fifty-five clinical and 30 nonclinical studies were reviewed. The evidence from randomized controlled trials evaluating the effectiveness of routine multimodality respiratory physiotherapy is conflicting. Physiotherapy that comprises early progressive mobilization has been shown to be feasible and safe, with data from randomized controlled trials demonstrating that it can improve function and shorten ICU and hospital length of stay. Available new evidence, published since 1999, suggests that physiotherapy intervention that comprises early progressive mobilization is beneficial for adult patients in the ICU in terms of its positive effect on functional ability and its potential to reduce ICU and hospital length of stay. These new findings suggest that early progressive mobilization should be implemented as a matter of priority in all adult ICUs and an area of clinical focus for ICU physiotherapists.


Potency of Michael reaction acceptors as inducers of enzymes that protect against carcinogenesis depends on their reactivity with sulfhydryl groups.
Induction of phase 2 enzymes and elevations of glutathione are major and sufficient strategies for protecting mammals and their cells against the toxic and carcinogenic effects of electrophiles and reactive forms of oxygen. Inducers belong to nine chemical classes and have few common properties except for their ability to modify sulfhydryl groups by oxidation, reduction, or alkylation. Much evidence suggests that the cellular "sensor" molecule that recognizes the inducers and signals the enhanced transcription of phase 2 genes does so by virtue of unique and highly reactive sulfhydryl functions that recognize and covalently react with the inducers. Benzylidene-alkanones and -cycloalkanones are Michael reaction acceptors whose inducer potency is profoundly increased by the presence of ortho- (but not other) hydroxyl substituent(s) on the aromatic ring(s). This enhancement correlates with more rapid reactivity of the ortho-hydroxylated derivatives with model sulfhydryl compounds. Proton NMR spectroscopy provides no evidence for increased electrophilicity of the beta-vinyl carbons (the presumed site of nucleophilic attack) on the hydroxylated inducers. Surprisingly, these ortho-hydroxyl groups display a propensity for extensive intermolecular hydrogen bond formation, which may raise the reactivity and facilitate addition of mercaptans, thereby raising inducer potencies.


pH-Dependent metal ion toxicity influences the antibacterial activity of two natural mineral mixtures.
Recent studies have demonstrated that several mineral products sold for medicinal purposes demonstrate antimicrobial activity, but little is known about the physicochemical properties involved in antibacterial activity. Using in vitro mineral suspension testing, we have identified two natural mineral mixtures, arbitrarily designated BY07 and CB07, with antibacterial activity against a broad-spectrum of bacterial pathogens. Mineral-derived aqueous leachates also exhibited antibacterial activity, revealing that chemical, not physical, mineral characteristics were responsible for the observed activity. The chemical properties essential for bactericidal activity against Escherichia coli were probed by testing antibacterial activity in the presence of metal chelators, the hydroxyl radical scavenger, thiourea, and varying pH levels. Chelation of the BY07 minerals with EDTA or desferrioxamine eliminated or reduced BY07 toxicity, respectively, suggesting a role of an acid-soluble metal species, particularly Fe(3+) or other sequestered metal cations, in mineral toxicity. This conclusion was supported by NMR relaxation data, which indicated that BY07 and CB07 leachates contained higher concentrations of chemically accessible metal ions than leachates from non-bactericidal mineral samples. We conclude that the acidic environment of the hydrated minerals significantly contributes to antibacterial activity by increasing the availability and toxicity of metal ions. These findings provide impetus for further investigation of the physiological effects of mineral products and their applications in complementary antibacterial therapies.


Spectral superresolution with ultrashort optical pulses.
A superresolution technique for the measurement of transmission, reflection, and absorption spectra is proposed. An ultrashort laser pulse is propagated in a dispersive element and then periodically phase modulated. The temporal modulation is transformed into periodic spectral modulation, for which the number of harmonics, 2M+1, is determined by the modulation index. The modulated pulse is transmitted through (reflected from) the sample to be tested and measured by a spectrometer. By performing 2M+1 measurements for 2M+1 delays between the dispersed pulse and modulation signal, one can restore the spectral response of the sample with superresolution after simple processing. We numerically demonstrate the measurement of the transmission spectrum of an ultranarrow optical filter with a minimum feature of 0.43 pm by an optical spectrum analyzer with a 10 pm resolution. A twentyfold enhancement of the resolution is achieved in the presence of noise with a level of 0.1%. The advantage of the system is its full reconfigurability.