International linkage of two food-borne hepatitis A clusters through traceback of mussels, the Netherlands, 2012.
This report describes an outbreak investigation starting with two closely related suspected food-borne clusters of Dutch hepatitis A cases, nine primary cases in total, with an unknown source in the Netherlands. The hepatitis A virus (HAV) genotype IA sequences of both clusters were highly similar (459/460 nt) and were not reported earlier. Food questionnaires and a case-control study revealed an association with consumption of mussels. Analysis of mussel supply chains identified the most likely production area. International enquiries led to identification of a cluster of patients near this production area with identical HAV sequences with onsets predating the first Dutch cluster of cases. The most likely source for this cluster was a case who returned from an endemic area in Central America, and a subsequent household cluster from which treated domestic sewage was discharged into the suspected mussel production area. Notably, mussels from this area were also consumed by a separate case in the United Kingdom sharing an identical strain with the second Dutch cluster. In conclusion, a small number of patients in a non-endemic area led to geographically dispersed hepatitis A outbreaks with food as vehicle. This link would have gone unnoticed without sequence analyses and international collaboration.


Rolling circle amplification: applications in nanotechnology and biodetection with functional nucleic acids.
Rolling circle amplification (RCA) is an isothermal, enzymatic process mediated by certain DNA polymerases in which long single-stranded (ss) DNA molecules are synthesized on a short circular ssDNA template by using a single DNA primer. A method traditionally used for ultrasensitive DNA detection in areas of genomics and diagnostics, RCA has been used more recently to generate large-scale DNA templates for the creation of periodic nanoassemblies. Various RCA strategies have also been developed for the production of repetitive sequences of DNA aptamers and DNAzymes as detection platforms for small molecules and proteins. In this way, RCA is rapidly becoming a highly versatile DNA amplification tool with wide-ranging applications in genomics, proteomics, diagnosis, biosensing, drug discovery, and nanotechnology.


A new view on the temperature insensitivity of intracavity SHG configuration.
Experimental results indicate that there is a distinct difference between the temperature tuning curves obtained with intracavity second harmonic generation (Intra-SHG) and extracavity SHG (Extra-SHG) configuration. By introducing the nonlinear SHG loss with the phase mismatch taken into consideration, the rate equations are established to describe the Intra-SHG laser performance. As for the widely used SHG crystal, KTP and LBO, theoretically calculated variation trends of the temperature insensitivity are in agreement with that obtained experimentally. The corresponding explanations have also been given in this paper. Our results may provide a new view on the temperature insensitivity of Intra-SHG configuration. The method employed in our study can be extended to other nonlinear crystals and intracavity nonlinear frequency conversions.


A model system to study antimicrobial strategies in endodontic biofilms.
The purpose of this work was to develop a model system to study antimicrobial strategies in endodontic biofilms. Enterococcus faecalis suspension was colonized in 10 human root canals. Five milliliters of Brain Heart Infusion (BHI) were mixed with 5 mL of the bacterial inoculums (E. faecalis) and inoculated with sufficient volume to fill the root canal during 60 days. This procedure was repeated every 72 h, always using 24-h pure culture prepared and adjusted to No. 1 MacFarland turbidity standard. Biofilm formation was analyzed by scanning electron microscopy (SEM). E. faecalis consistently adhered to collagen structure, colonized dentin surface, progressed towards the dentinal tubules and formed a biofilm. The proposed biofilm model seems to be viable for studies on antimicrobial strategies, and allows for a satisfactory colonization time of selected bacterial species with virulence and adherence properties.


Laboratory practices for prenatal group B streptococcal screening--seven states, 2003.
In the United States, group B streptococcus (GBS) is the leading cause of serious bacterial infections in newborns. In 1996, consensus guidelines for use of intrapartum antibiotic prophylaxis (IAP) to prevent perinatal GBS disease recommended either of two methods for identifying candidates for chemoprophylaxis: 1) late prenatal culture-based screening for GBS colonization or 2) monitoring of women intrapartum for particular risk factors associated with early-onset GBS disease. Evidence that culture-based screening was substantially more effective than the risk-based approach led to revised guidelines in 2002 recommending late prenatal GBS screening for all pregnant women. Although methods for isolation and identification of GBS from prenatal specimens remained the same as those recommended in 1996, the 2002 guidelines recommended that laboratories perform antimicrobial susceptibility testing on prenatal GBS isolates from women at high risk for penicillin anaphylaxis and clarified that laboratories should report the presence of any GBS in urine specimens from pregnant women. To assess laboratory adherence to recommendations in the 2002 guidelines, CDC's Active Bacterial Core surveillance (ABCs)/Emerging Infections Program Network surveyed clinical laboratories about prenatal culture-processing practices in 2003. This report summarizes the results of that survey, which indicated that, although adherence to GBS isolation procedures was high, opportunities exist to improve implementation of recommendations related to antimicrobial susceptibility testing and GBS bacteriuria.


Detox diets for toxin elimination and weight management: a critical review of the evidence.
Detox diets are popular dieting strategies that claim to facilitate toxin elimination and weight loss, thereby promoting health and well-being. The present review examines whether detox diets are necessary, what they involve, whether they are effective and whether they present any dangers. Although the detox industry is booming, there is very little clinical evidence to support the use of these diets. A handful of clinical studies have shown that commercial detox diets enhance liver detoxification and eliminate persistent organic pollutants from the body, although these studies are hampered by flawed methodologies and small sample sizes. There is preliminary evidence to suggest that certain foods such as coriander, nori and olestra have detoxification properties, although the majority of these studies have been performed in animals. To the best of our knowledge, no randomised controlled trials have been conducted to assess the effectiveness of commercial detox diets in humans. This is an area that deserves attention so that consumers can be informed of the potential benefits and risks of detox programmes.


Modulation of experimental allergic encephalomyelitis with anti-T suppressor factor antibodies.
mAb reactive with T suppressor factors (TsF) were used to alter the course of myelin basic protein-induced experimental allergic encephalomyelitis in (SJL/J x PL/J)F1 mice. In vivo administration of mAb 14-12, reactive with effector TsF, exacerbated the clinical expression of encephalomyelitis as evidenced by prolonged periods of total limb paralysis in affected animals. This aggravation of disease signs is probably related to the inhibition of effector Ts function by mAb 14-12 thus allowing T cell autoreactivity to proceed unchecked. Disease course was influenced more favorably by i.v. administration of mAb 14-30 reactive with a subset of inducer TsF. Ten days of treatment with this mAb resulted in a reduction in the incidence and severity of disease, noted as the development of minimal limb weakness but no paralysis in the majority of affected animals. Adoptive transfer experiments revealed the presence of Ag-specific Ts in mAb 14-30-treated mice that inhibited recipient Lyt-1+ responses to myelin basic protein, the immunizing autoantigen. Suppression by transferred Ts was revealed only by treatment of the donor population with anti-Lyt-1.2 plus C, however, indicating a role for contrasuppressor activity in the regulation of autoimmune T cell function. Results are considered relevant to the potential for immunotherapeutic management of multiple sclerosis in man.


The development of print tuning in children with dyslexia: evidence from longitudinal ERP data supported by fMRI.
A consistent finding in functional brain imaging studies of reading with dyslexia is reduced inferior occipito-temporal activation linked to deviant processing of visual word forms. Time-sensitive event-related potentials (ERP) further revealed reduced inferior occipito-temporal N1 tuning for print in dyslexic 2nd graders suggesting the reduction affects fast processing and the initial development of dyslexia. Here, we followed up the same groups with ERP recordings and investigated how fast print tuning deficits in dyslexia develop from 2nd to 5th grade. Using functional magnetic resonance imaging (fMRI), we further characterized spatial aspects of print tuning in the 5th grade. The robust N1 tuning deficit for print in the dyslexic 2nd graders had largely disappeared by grade 5 consistent with a developmental delay. Reduced word-specific activation in dyslexic 5th grader's fMRI data occurred bilaterally in middle temporal regions and in the left posterior superior sulcus. Although no group differences in inferior occipito-temporal regions appeared in the whole brain analysis, a region of interest analysis of the Visual Word Form Area revealed that control children showed a more lateralized word-specific activation pattern than the children with dyslexia. The results suggest that while impaired N1 tuning for print plays a major role for dyslexia at the beginning of learning to read, other aspects of visual word form processing in the same region remain impaired in dyslexic children after several years of reading practice. Overall, neural deficits associated with dyslexia appear to be plastic and to change throughout development and reading acquisition.


Citrobacter freundii fitness during bloodstream infection.
Sepsis resulting from microbial colonization of the bloodstream is a serious health concern associated with high mortality rates. The objective of this study was to define the physiologic requirements of Citrobacter freundii in the bloodstream as a model for bacteremia caused by opportunistic Gram-negative pathogens. A genetic screen in a murine host identified 177 genes that contributed significantly to fitness, the majority of which were broadly classified as having metabolic or cellular maintenance functions. Among the pathways examined, the Tat protein secretion system conferred the single largest fitness contribution during competition infections and a putative Tat-secreted protein, SufI, was also identified as a fitness factor. Additional work was focused on identifying relevant metabolic pathways for bacteria in the bloodstream environment. Mutations that eliminated the use of glucose or mannitol as carbon sources in vitro resulted in loss of fitness in the murine model and similar results were obtained upon disruption of the cysteine biosynthetic pathway. Finally, the conservation of identified fitness factors was compared within a cohort of Citrobacter bloodstream isolates and between Citrobacter and Serratia marcescens, the results of which suggest the presence of conserved strategies for bacterial survival and replication in the bloodstream environment.


Senior dental students' career intentions, work-life balance and retirement plans.
To gather information from senior dental students about their future career plans, with particular emphasis on work-life balance issues, their attitudes towards the NHS and retirement plans. Senior dental students at the Universities of Dundee and Manchester were asked to complete a voluntary anonymous questionnaire. In all 141 questionnaires were completed, 42 by students in Manchester and 114 in Dundee. On qualification nearly all surveyed intend to work full time but after five years one quarter (26%) of females intend to work part time. This is significantly (p < 0.05) different from males where nearly all (98%) intend to work full time. Although the majority (65%) intend to work in general practice significant numbers (19%) wish to have a career in hospital dentistry and very few (3%) in community dentistry. Senior students seem to show no more commitment to the NHS than those in our previous study of dental school applicants. Only 3% intend to work exclusively for the NHS and 18% intend to work exclusively in the private sector. Surprising numbers had plans to retire or go part time before 60 years of age. Only 20% of the sample intended to continue working full time after the age of 60 years. The mode age that those surveyed intended to start a family was 30 years and a large majority of both sexes thought this would interrupt their professional life. More than half of the sample intend to take time out of dentistry until their children attended primary school (female 63%, male 38%) and 6% (female 6%, male 8%) until secondary school. Many of our findings suggest that future generations of dentists may have a pattern of professional life that will have the effect of reducing their clinical commitment, although it is not possible to determine how significant an effect this will have on the workforce. It may, however, be appropriate to take career intention into account when workforce planning.


Infrared absorption spectrum of the simplest deuterated Criegee intermediate CD2OO.
We report a transient infrared (IR) absorption spectrum of the simplest deuterated Criegee intermediate CD2OO recorded using a step-scan Fourier-transform spectrometer coupled with a multipass absorption cell. CD2OO was produced from photolysis of flowing mixtures of CD2I2, N2, and O2 (13 or 87 Torr) with laser light at 308 nm. The recorded spectrum shows close structural similarity with the spectrum of CH2OO reported previously [Y.-T. Su et al., Science 340, 174 (2013)]. The four bands observed at 852, 1017, 1054, and 1318 cm(-1) are assigned to the OO stretching mode, two distinct in-plane OCD bending modes, and the CO stretching mode of CD2OO, respectively, according to vibrational wavenumbers, IR intensities, rotational contours, and deuterium-isotopic shifts predicted with extensive quantum-chemical calculations. The CO-stretching mode of CD2OO at 1318 cm(-1) is blue shifted from the corresponding band of CH2OO at 1286 cm(-1); this can be explained by a mechanism based on mode mixing and isotope substitution. A band near 936 cm(-1), observed only at higher pressure (87 Torr), is tentatively assigned to the CD2 wagging mode of CD2IOO.


Factors affecting bilirubin levels during first 48 hours of life in healthy infants.
To investigate the relationship of delivery type, maternal anesthesia, feeding modalities, and first feeding and meconium passage times with early bilirubin levels of healthy infants. Cord, 24 hours' and 48 hours' total bilirubin levels were measured in 388 study infants. Infants born with cesarean section were fed later and more often had mixed feeding. First meconium passage was delayed with general anesthesia. Cord, 24 and 48 hours' bilirubin levels were not correlated with first feeding time, meconium passage time, mode of delivery, existence and type of anesthesia, and feeding modalities. Being in high intermediate risk zone at 72 hours of Bhutani's nomogram was only related to first feeding time and high cord bilirubin level. Late preterm infants were more frequently born with cesarean section and offered supplementary formula. Therefore, first meconium passage times and bilirubin levels were similar in the late preterm and term infants. Type of delivery or anesthesia, late prematurity, feeding modalities, and first meconium passage time were not related to early bilirubin levels in healthy neonates, but delayed first feeding and high cord bilirubin levels were related to be in higher risk zone for later hyperbilirubinemia.


Stimulation of a subset of normal resting T lymphocytes by a monoclonal antibody to a crossreactive determinant of the human T cell antigen receptor.
A previous study from this laboratory described a monoclonal antibody, S511, that reacted with the T cell antigen receptor on a human T cell leukemia and also on 1-2% of circulating T lymphocytes in all normal individuals tested. The data presented in the present study demonstrate that, when normal T lymphocytes are cultured with or without irradiated non-T cells in the presence of soluble S511 antibody, a concentration- and time-dependent proliferation of the S511-reactive population occurred. Proliferation indices as high as 184 times greater than control were observed, which represents a major stimulatory effect on the initially minor S511+ subset. When S511+ cells were studied for evidence of prior activation, they were shown to be unresponsive to interleukin 2 (IL-2) unless exposed to S511 antibody, and were shown to be in the G0/G1 phase of the cell cycle. Thus, the S511 antibody activated resting normal T cells in a manner analogous to specific antigen binding to the T cell antigen receptor. The leukemic S511 molecule has been shown previously to differ from most other antigen receptors in the mobility of the two chains at 43 and 38 kD and the neutral isoelectric point of each chain. Expansion of reactive normal cells by S511-Sepharose permitted the development of IL-2-dependent T cell lines enriched for S511-bearing cells. The antigen receptor molecules on one such polyclonal S511-enriched T cell line were immunoprecipitated with S511 antibody and shown to have comparable mobility to that present on the leukemic cells, but to possess a greater heterogeneity of mobility. Thus, the leukemic cells and normal cells express similar T cell receptor molecules. The differences in the S511 T cell antigen receptor molecule possibly relate to differences in glycosylation or polypeptide structure.


Trends and Demographic Disparities in Diabetes Hospital Admissions: Analyses of Serial Cross-Sectional National and State Data, 2008-2017.
To analyze national and state-specific trends in diabetes-related hospital admissions and determine whether disparities in rates of admission exist between demographic groups and geographically dispersed states. We conducted serial cross-sectional analyses of the National Inpatient Sample (2008, 2011, 2014, and 2016) and State Inpatient Databases for Arizona, Florida, Kentucky, Iowa, Maryland, Nebraska, New Jersey, New York, North Carolina, Utah, and Vermont for 2008, 2011, 2014, and 2016/2017 among adult patients with type 1 and type 2 diabetes-related ICD codes (ICD-9 [250.XX] or ICD-10 [E10.XXX, E11.XXX, and E13.XXX]. We measured hospitalization rates for people with diabetes (all-cause hospitalizations) and for admissions with a primary diagnosis of diabetes or diabetes-related complications (diabetes-specific hospitalizations) per 10,000 people per year. Nationally, all-cause and diabetes-specific hospitalizations declined by 3.1% (95% CI -5.5, -0.7) and 19.1% (95% CI -21.6, -16.6), respectively, over 2008 to 2016. The analysis of individual states showed that diabetes-specific admissions in individuals ≥65 years old declined during this time (16.3-48.8% decrease) but increased among patients 18-29 years old (10.5-81.5% increase) and that rural diabetes-specific admissions decreased in just over half of the included states (15.2-69.2% decrease). There were no differences in changes in admission rates among different racial/ethnic groups. Overall, rates of diabetes-related hospitalizations decreased over 2008 to 2016/2017, but there were large state-level differences across subgroups of patients. The rise in diabetes hospitalizations among young adults is a cause for concern. These state- and subpopulation-level differences highlight the need for state-level policies and interventions to address disparities in diabetes health care use.


Double-edge molecular measurement of lidar wind profiles at 355 nm.
We built a direct-detection Doppler lidar based on the double-edge molecular technique and made the what we believe to be the first molecular-based wind measurements using the eye-safe 355-nm wavelength. Three etalon bandpasses are obtained with step etalons on a single pair of etalon plates. We eliminate long-term frequency drift of the laser and the capacitively stabilized etalon by locking the etalon to the laser frequency. We use a low-angle design to avoid polarization effects. Wind measurements of 1-2-m /s accuracy are obtained to 10-km altitude with 5 mJ of laser energy, a 750-s integration, and a 25-cm telescope. Good agreement is obtained between lidar and rawinsonde measurements.


The effect of therapeutic exercise on activation of the deep cervical flexor muscles in people with chronic neck pain.
Deep cervical flexor muscle (DCF) activation is impaired with neck pain. This study investigated the effects of low load cranio-cervical flexion (C-CF) and neck flexor strengthening exercises on spatial and temporal characteristics of DCF activation during a neck movement task and a task challenging the neck's postural stability. Forty-six chronic neck pain subjects were randomly assigned to an exercise group and undertook a 6-week training program. Electromyographic (EMG) activity was recorded from the DCF, sternocleidomastoid (SCM) and anterior scalene (AS) muscles pre and post intervention during the cranio-cervical flexion test (CCFT) and during perturbations induced by rapid, unilateral shoulder flexion and extension. C-CF training increased DCF EMG amplitude and decreased SCM and AS EMG amplitude across all stages of the CCFT (all P<0.05). No change occurred in DCF EMG amplitude following strength training. There was no significant between group difference in pre-post intervention change in relative latency of DCF but a greater proportion of the C-CF group shortened the relative latency between the activation of the deltoid and the DCF during rapid arm movement compared to the strength group (P<0.05). Specific low load C-CF exercise changes spatial and temporal characteristics of DCF activation which may partially explain its efficacy in rehabilitation.


iHam and pyHam: visualizing and processing hierarchical orthologous groups.
The evolutionary history of gene families can be complex due to duplications and losses. This complexity is compounded by the large number of genomes simultaneously considered in contemporary comparative genomic analyses. As provided by several orthology databases, hierarchical orthologous groups (HOGs) are sets of genes that are inferred to have descended from a common ancestral gene within a species clade. This implies that the set of HOGs defined for a particular clade correspond to the ancestral genes found in its last common ancestor. Furthermore, by keeping track of HOG composition along the species tree, it is possible to infer the emergence, duplications and losses of genes within a gene family of interest. However, the lack of tools to manipulate and analyse HOGs has made it difficult to extract, display and interpret this type of information. To address this, we introduce interactive HOG analysis method, an interactive JavaScript widget to visualize and explore gene family history encoded in HOGs and python HOG analysis method, a python library for programmatic processing of genes families. These complementary open source tools greatly ease adoption of HOGs as a scalable and interpretable concept to relate genes across multiple species. iHam's code is available at https://github.com/DessimozLab/iHam or can be loaded dynamically. pyHam's code is available at https://github.com/DessimozLab/pyHam and or via the pip package 'pyham'.


Drinking hydrogen water ameliorated cognitive impairment in senescence-accelerated mice.
Hydrogen has been reported to have neuron protective effects due to its antioxidant properties, but the effects of hydrogen on cognitive impairment due to senescence-related brain alterations and the underlying mechanisms have not been characterized. In this study, we investigated the efficacies of drinking hydrogen water for prevention of spatial memory decline and age-related brain alterations using senescence-accelerated prone mouse 8 (SAMP8), which exhibits early aging syndromes including declining learning ability and memory. However, treatment with hydrogen water for 30 days prevented age-related declines in cognitive ability seen in SAMP8 as assessed by a water maze test and was associated with increased brain serotonin levels and elevated serum antioxidant activity. In addition, drinking hydrogen water for 18 weeks inhibited neurodegeneration in hippocampus, while marked loss of neurons was noted in control, aged brains of mice receiving regular water. On the basis of our results, hydrogen water merits further investigation for possible therapeutic/preventative use for age-related cognitive disorders.


[Cecal amelanotic melanoma].
A 73 year old male patient who presented to emergency with severe anemia (Hb 4.9 g /dL), referring to a month with dark stools, fatigue and significant weight loss. The high endoscopic evaluation found no potentially bleeding lesions. Subsequently, we confirmed the presence of red blood in stools, like enterorrhagia and underwent a colonoscopy, in which two elevated lesions were found in the cecum: a pedunculated (with active bleeding, oozing) and other sessile; both were removed. the pathology showed that corresponded to amelanotic melanoma in cecal region. Physical examination revealed no malignant skin lesions. Recurrent gastrointestinal bleeding forced to make another colonoscopy that showed new bleeding lesion of the cecum. He was operated with a cecostomy and cecal excision of the lesion.. The tomography scan showed metastases in mediastinum and surgical implants were found in liver metastases and small intestine. The patient continued treatment in medical oncology. We report this case, because gastrointestinal bleeding is an unusual clinical presentation of malignant melanoma.


Crosstalk between epigenetics and metabolism-Yin and Yang of histone demethylases and methyltransferases in cancer.
Histone methylation is an epigenetic modification of chromatin undergoing dynamic changes and balancing tissue-specific demands of proliferation and differentiation. In cancer, aberrant histone methylation can facilitate oncogenic and tumor suppression programs by modulating gene expression. Histone remodelers such as lysine methyltransferases and lysine demethylases are seemingly opposite or contrary forces but may be part of an interconnected network complementing each other. We identify several layers of molecular communication where epigenetic master regulators engage in crosstalk between tumor metabolism and histone remodeling. Epigenetic master regulators have the ability to cooperate with members of the transcriptional machinery, DNA methyltransferases, as well as other histone modifiers. High-throughput sequencing and omics data in combination with cancer systems biology analysis have the power to prioritize regulatory events epigenome-wide.


Quantum simulation of decoherence in optical waveguide lattices.
We suggest that propagation of nonclassical light in lattices of optical waveguides can provide a laboratory tool to simulate quantum decoherence phenomena with high non-Markovian features. As examples, we study decoherence of optical Schrödinger cats in a lattice that mimics a dissipative quantum harmonic oscillator coupled to a quantum bath, showing fractional decoherence in the strong coupling regime, and Bloch oscillations of optical Schrödinger cats, where damped revivals of the coherence can be observed.


HIV-1 immune activation induces Siglec-1 expression and enhances viral trans-infection in blood and tissue myeloid cells.
Myeloid cells are key players in the recognition and response of the host against invading viruses. Paradoxically, upon HIV-1 infection, myeloid cells might also promote viral pathogenesis through trans-infection, a mechanism that promotes HIV-1 transmission to target cells via viral capture and storage. The receptor Siglec-1 (CD169) potently enhances HIV-1 trans-infection and is regulated by immune activating signals present throughout the course of HIV-1 infection, such as interferon α (IFNα). Here we show that IFNα-activated dendritic cells, monocytes and macrophages have an enhanced ability to capture and trans-infect HIV-1 via Siglec-1 recognition of viral membrane gangliosides. Monocytes from untreated HIV-1-infected individuals trans-infect HIV-1 via Siglec-1, but this capacity diminishes after effective antiretroviral treatment. Furthermore, Siglec-1 is expressed on myeloid cells residing in lymphoid tissues, where it can mediate viral trans-infection. Siglec-1 on myeloid cells could fuel novel CD4(+) T-cell infections and contribute to HIV-1 dissemination in vivo.


Effects of Standard-Dose Prophylactic, High-Dose Prophylactic, and Therapeutic Anticoagulation in Patients With Hypoxemic COVID-19 Pneumonia: The ANTICOVID Randomized Clinical Trial.
Given the high risk of thrombosis and anticoagulation-related bleeding in patients with hypoxemic COVID-19 pneumonia, identifying the lowest effective dose of anticoagulation therapy for these patients is imperative. To determine whether therapeutic anticoagulation (TA) or high-dose prophylactic anticoagulation (HD-PA) decreases mortality and/or disease duration compared with standard-dose prophylactic anticoagulation (SD-PA), and whether TA outperforms HD-PA; and to compare the net clinical outcomes among the 3 strategies. The ANTICOVID randomized clinical open-label trial included patients with hypoxemic COVID-19 pneumonia requiring supplemental oxygen and having no initial thrombosis on chest computer tomography with pulmonary angiogram at 23 health centers in France from April 14 to December 13, 2021. Of 339 patients randomized, 334 were included in the primary analysis-114 patients in the SD-PA group, 110 in the HD-PA, and 110 in the TA. At randomization, 90% of the patients were in the intensive care unit. Data analyses were performed from April 13, 2022, to January 3, 2023. Patients were randomly assigned (1:1:1) to receive either SD-PA, HD-PA, or TA with low-molecular-weight or unfractionated heparin for 14 days. A hierarchical criterion of all-cause mortality followed by time to clinical improvement at day 28. Main secondary outcome was net clinical outcome at day 28 (composite of thrombosis, major bleeding, and all-cause death). Among the study population of 334 individuals (mean [SD] age, 58.3 [13.0] years; 226 [67.7%] men and 108 [32.3%] women), use of HD-PA and SD-PA had similar probabilities of favorable outcome (47.3% [95% CI, 39.9% to 54.8%] vs 52.7% [95% CI, 45.2% to 60.1%]; P = .48), as did TA compared with SD-PA (50.9% [95% CI, 43.4% to 58.3%] vs 49.1% [95% CI, 41.7% to 56.6%]; P = .82) and TA compared with HD-PA (53.5% [95% CI 45.8% to 60.9%] vs 46.5% [95% CI, 39.1% to 54.2%]; P = .37). Net clinical outcome was met in 29.8% of patients receiving SD-PA (20.2% thrombosis, 2.6% bleeding, 14.0% death), 16.4% receiving HD-PA (5.5% thrombosis, 3.6% bleeding, 11.8% death), and 20.0% receiving TA (5.5% thrombosis, 3.6% bleeding, 12.7% death). Moreover, HD-PA and TA use significantly reduced thrombosis compared with SD-PA (absolute difference, -14.7 [95% CI -6.2 to -23.2] and -14.7 [95% CI -6.2 to -23.2], respectively). Use of HD-PA significantly reduced net clinical outcome compared with SD-PA (absolute difference, -13.5; 95% CI -2.6 to -24.3). This randomized clinical trial found that compared with SD-PA, neither HD-PA nor TA use improved the primary hierarchical outcome of all-cause mortality or time to clinical improvement in patients with hypoxemic COVID-19 pneumonia; however, HD-PA resulted in significantly better net clinical outcome by decreasing the risk of de novo thrombosis. ClinicalTrials.gov Identifier: NCT04808882.


Machine learning models in electronic health records can outperform conventional survival models for predicting patient mortality in coronary artery disease.
Prognostic modelling is important in clinical practice and epidemiology for patient management and research. Electronic health records (EHR) provide large quantities of data for such models, but conventional epidemiological approaches require significant researcher time to implement. Expert selection of variables, fine-tuning of variable transformations and interactions, and imputing missing values are time-consuming and could bias subsequent analysis, particularly given that missingness in EHR is both high, and may carry meaning. Using a cohort of 80,000 patients from the CALIBER programme, we compared traditional modelling and machine-learning approaches in EHR. First, we used Cox models and random survival forests with and without imputation on 27 expert-selected, preprocessed variables to predict all-cause mortality. We then used Cox models, random forests and elastic net regression on an extended dataset with 586 variables to build prognostic models and identify novel prognostic factors without prior expert input. We observed that data-driven models used on an extended dataset can outperform conventional models for prognosis, without data preprocessing or imputing missing values. An elastic net Cox regression based with 586 unimputed variables with continuous values discretised achieved a C-index of 0.801 (bootstrapped 95% CI 0.799 to 0.802), compared to 0.793 (0.791 to 0.794) for a traditional Cox model comprising 27 expert-selected variables with imputation for missing values. We also found that data-driven models allow identification of novel prognostic variables; that the absence of values for particular variables carries meaning, and can have significant implications for prognosis; and that variables often have a nonlinear association with mortality, which discretised Cox models and random forests can elucidate. This demonstrates that machine-learning approaches applied to raw EHR data can be used to build models for use in research and clinical practice, and identify novel predictive variables and their effects to inform future research.


Transcription factor induction of human oligodendrocyte progenitor fate and differentiation.
Human oligodendrocyte progenitor cell (OPC) specification and differentiation occurs slowly and limits the potential for cell-based treatment of demyelinating disease. In this study, using FACS-based isolation and microarray analysis, we identified a set of transcription factors expressed by human primary CD140a(+)O4(+) OPCs relative to CD133(+)CD140a(-) neural stem/progenitor cells (NPCs). Among these, lentiviral overexpression of transcription factors ASCL1, SOX10, and NKX2.2 in NPCs was sufficient to induce Sox10 enhancer activity, OPC mRNA, and protein expression consistent with OPC fate; however, unlike ASCL1 and NKX2.2, only the transcriptome of SOX10-infected NPCs was induced to a human OPC gene expression signature. Furthermore, only SOX10 promoted oligodendrocyte commitment, and did so at quantitatively equivalent levels to native OPCs. In xenografts of shiverer/rag2 animals, SOX10 increased the rate of mature oligodendrocyte differentiation and axon ensheathment. Thus, SOX10 appears to be the principle and rate-limiting regulator of myelinogenic fate from human NPCs.


The association of bone attrition with knee pain and other MRI features of osteoarthritis.
To determine whether bone attrition (flattening or depression of the subchondral bone) was associated with the presence and severity of knee pain and to evaluate the coexistence of attrition and other MRI features likely associated with pain. Participants in the Framingham Osteoarthritis Study, a community cohort unselected for OA, answered questions about knee pain and underwent knee x rays and MRI. Attrition, bone marrow lesions (BMLs) and effusions were scored on MRI using the WORMS scale. We assessed attrition in knees with and without pain, and using logistic regression examined its association with pain adjusting for age, gender, Kellgren-Lawrence (K-L)grade, BMI, BML and effusion. We also explored the relation between attrition, pain severity and nocturnal pain. Attrition (Grade >2) was present in 28% (167/592) of painful knees and in 10% (106/1035) of nonpainful knees (adjusted OR 1.6 (95% CI 1.1 to 2.2)). Of knees with OA (n=368), 74% had pain if attrition was present and 58% if it was absent (adjusted OR 1.2 (95%CI 0.7 to 2.0)). Of knees without OA (n=1222), pain was reported in 39% of knees with attrition and in 27%without it (adjusted OR 2.1 (95% CI 1.1 to 4.0)). We found no association between either attrition/pain severity or attrition/nocturnal pain. Attrition often co-occurred with other OA features associated with pain such as BMLs and effusions. Attrition was associated independently with knee pain. Unlike knees without OA, the association was lost in OA knees where other pathological features that may cause pain also coexisted.


The effects of nutritional supplements on the symptoms of fibromyalgia and chronic fatigue syndrome.
This article reports the results of a within-subject design. Fifty subjects with a physician diagnosis of fibromyalgia (FM) and/or chronic fatigue syndrome (CFS) were interviewed using a structured interview from. Each subject was interviewed initially, and again nine months later (follow-up). Subjects had, on their own, consumed nutritional supplements including freeze-dried aloe vera gel extract; a combination of freeze-dried aloe vera gel extract and additional plant-derived saccharides; freeze-dried fruits and vegetables in combination with the saccharides; and a formulation of dioscorea complex containing the saccharides and a vitamin/mineral complex. With medical treatments, approximately 25 percent of FM patients improve, but the beneficial effects of medical treatment rarely persist more than a few months. All subjects in this study had received some form of medical treatment prior to taking the nutritional supplements, but none with enduring success. Nutritional supplements resulted in a remarkable reduction in initial symptom severity, with continued improvement in the period between initial assessment and the follow-up. Further research is needed to verify these results, specifically crossover designs in well-defined populations.


Enzymatic Modification of N-Terminal Proline Residues Using Phenol Derivatives.
A convenient enzymatic strategy is reported for the modification of proline residues in the N-terminal positions of proteins. Using a tyrosinase enzyme isolated from Agaricus bisporus (abTYR), phenols and catechols are oxidized to highly reactive o-quinone intermediates that then couple to N-terminal proline residues in high yield. Key advantages of this bioconjugation method include (1) the use of air-stable precursors that can be prepared on large scale if needed, (2) mild reaction conditions, including low temperatures, (3) the targeting of native functional groups that can be introduced readily on most proteins, and (4) the use of molecular oxygen as the sole oxidant. This coupling strategy was successfully demonstrated for the attachment of a variety of phenol-derivatized cargo molecules to a series of protein substrates, including self-assembled viral capsids, enzymes, and a chitin binding domain (CBD). The ability of the CBD to bind to the surfaces of yeast cells was found to be unperturbed by this modification reaction.


Fcgamma receptor-like activity of hepatitis C virus core protein.
We have previously demonstrated that viral particles with the properties of nonenveloped hepatitis C virus (HCV) nucleocapsids occur in the serum of HCV-infected individuals (1). We show here that nucleocapsids purified directly from serum or isolated from HCV virions have FcgammaR-like activity and bind "nonimmune" IgG via its Fcgamma domain. HCV core proteins produced in Escherichia coli and in the baculovirus expression system also bound "nonimmune" IgG and their Fcgamma fragments. Folded conformation was required for IgG binding because the FcgammaR-like site of the core protein was inactive in denaturing conditions. Studies with synthetic core peptides showed that the region spanning amino acids 3-75 was essential for formation of the IgG-binding site. The interaction between the HCV core and human IgG is more efficient in acidic (pH 6.0) than in neutral conditions. The core protein-binding site on the IgG molecule differs from those for C1q, FcgammaRII (CD32), and FcgammaRIII (CD16) but overlaps with that for soluble protein A from Staphylococcus aureus (SpA), which is located in the CH2-CH3 interface of IgG. These characteristics of the core-IgG interaction are very similar to those of the neonatal FcRn. Surface plasmon resonance studies suggested that the binding of an anti-core antibody to HCV core protein might be "bipolar" through its paratope to the corresponding epitope and by its Fcgamma region to the FcgammaR-like motif on this protein. These features of HCV nucleocapsids and HCV core protein may confer an advantage for HCV in terms of survival by interfering with host defense mechanisms mediated by the Fcgamma part of IgG.


The Impact of Lumbar Spinal Stenosis, Knee Osteoarthritis, and Loss of Lumbar Lordosis on the Quality of Life: Findings from the Katsuragi Low Back Pain Study.
Musculoskeletal diseases and spinal malalignment are associated with poorer quality of life (QOL) in the elderly. However, to date, few general population cohort studies have focused on these conditions together. Our objectives were to clarify the associations between musculoskeletal degenerative diseases and/or spinal malalignment with QOL measures in a group of Japanese older adults. In this cross-sectional study, we analyzed data from 334 individuals recruited from the local population (120 men, 214 women; mean age 62.7 years; range 40-75). Low back pain (LBP) was assessed by questionnaire, and lumbar spinal stenosis (LSS) was diagnosed using a validated lumbar spinal stenosis support tool. Knee osteoarthritis (KOA) was diagnosed by the presence of clinical knee pain plus radiographic KOA. Spinal radiographs were used to assess the degree of lumbar lordosis (LL) and sagittal vertical alignment (SVA). QOL assessment was performed using the Oswestry Disability Index (ODI). A score of 12 was used as a cut-off point for poor QOL. Overall, 107 (32.0%) participants had an ODI > 12 (cases), and the remaining 227 individuals were designated controls. LBP, LSS, KOA, and LL were associated with poorer QOL, both in basic models and models adjusted for age, sex, and BMI. Associations persisted after adjustment for other musculoskeletal outcomes. In a free-living Japanese population, the poor QOL odds are increased by LBP, LSS, KOA, and certain spinal radiographic features, loss of LL, and increased SVA. Poor QOL odds were greatest in those diagnosed with LSS or KOA. From spinal radiographs, decreased LL and increased SVA were also predictors of poor QOL.


Incorporating functional inter-relationships into protein function prediction algorithms.
Functional classification schemes (e.g. the Gene Ontology) that serve as the basis for annotation efforts in several organisms are often the source of gold standard information for computational efforts at supervised protein function prediction. While successful function prediction algorithms have been developed, few previous efforts have utilized more than the protein-to-functional class label information provided by such knowledge bases. For instance, the Gene Ontology not only captures protein annotations to a set of functional classes, but it also arranges these classes in a DAG-based hierarchy that captures rich inter-relationships between different classes. These inter-relationships present both opportunities, such as the potential for additional training examples for small classes from larger related classes, and challenges, such as a harder to learn distinction between similar GO terms, for standard classification-based approaches. We propose a method to enhance the performance of classification-based protein function prediction algorithms by addressing the issue of using these interrelationships between functional classes constituting functional classification schemes. Using a standard measure for evaluating the semantic similarity between nodes in an ontology, we quantify and incorporate these inter-relationships into the k-nearest neighbor classifier. We present experiments on several large genomic data sets, each of which is used for the modeling and prediction of over hundred classes from the GO Biological Process ontology. The results show that this incorporation produces more accurate predictions for a large number of the functional classes considered, and also that the classes benefitted most by this approach are those containing the fewest members. In addition, we show how our proposed framework can be used for integrating information from the entire GO hierarchy for improving the accuracy of predictions made over a set of base classes. Finally, we provide qualitative and quantitative evidence that this incorporation of functional inter-relationships enables the discovery of interesting biology in the form of novel functional annotations for several yeast proteins, such as Sna4, Rtn1 and Lin1. We implemented and evaluated a methodology for incorporating interrelationships between functional classes into a standard classification-based protein function prediction algorithm. Our results show that this incorporation can help improve the accuracy of such algorithms, and help uncover novel biology in the form of previously unknown functional annotations. The complete source code, a sample data set and the additional files for this paper are available free of charge for non-commercial use at http://www.cs.umn.edu/vk/gaurav/functionalsimilarity/.


EuPaGDT: a web tool tailored to design CRISPR guide RNAs for eukaryotic pathogens.
Recent development of CRISPR-Cas9 genome editing has enabled highly efficient and versatile manipulation of a variety of organisms and adaptation of the CRISPR-Cas9 system to eukaryotic pathogens has opened new avenues for studying these otherwise hard to manipulate organisms. Here we describe a webtool, Eukaryotic Pathogen gRNA Design Tool (EuPaGDT; available at http://grna.ctegd.uga.edu), which identifies guide RNA (gRNA) in input gene(s) to guide users in arriving at well-informed and appropriate gRNA design for many eukaryotic pathogens. Flexibility in gRNA design, accommodating unique eukaryotic pathogen (gene and genome) attributes and high-throughput gRNA design are the main features that distinguish EuPaGDT from other gRNA design tools. In addition to employing an array of known principles to score and rank gRNAs, EuPaGDT implements an effective on-target search algorithm to identify gRNA targeting multi-gene families, which are highly represented in these pathogens and play important roles in host-pathogen interactions. EuPaGDT also identifies and scores microhomology sequences flanking each gRNA targeted cut-site; these sites are often essential for the microhomology-mediated end joining process used for double-stranded break repair in these organisms. EuPaGDT also assists users in designing single-stranded oligonucleotides for homology directed repair. In batch processing mode, EuPaGDT is able to process genome-scale sequences, enabling preparation of gRNA libraries for large-scale screening projects.


Semiconducting black phosphorus: synthesis, transport properties and electronic applications.
Phosphorus is one of the most abundant elements preserved in earth, and it comprises a fraction of ∼0.1% of the earth crust. In general, phosphorus has several allotropes, and the two most commonly seen allotropes, i.e. white and red phosphorus, are widely used in explosives and safety matches. In addition, black phosphorus, though rarely mentioned, is a layered semiconductor and has great potential in optical and electronic applications. Remarkably, this layered material can be reduced to one single atomic layer in the vertical direction owing to the van der Waals structure, and is known as phosphorene, in which the physical properties can be tremendously different from its bulk counterpart. In this review article, we trace back to the research history on black phosphorus of over 100 years from the synthesis to material properties, and extend the topic from black phosphorus to phosphorene. The physical and transport properties are highlighted for further applications in electronic and optoelectronics devices.


Instability due to unrecognised fracture-subluxations after apparently isolated injuries of the cervical spine.
Instability may present at a different level after successful stabilisation of an unstable segment in apparently isolated injuries of the cervical spine. It can give rise to progressive deformity or symptoms which require further treatment. We performed one or more operations for unstable cervical spinal injuries on 121 patients over a period of 90 months. Of these, five were identified as having instability due to an initially unrecognised fracture-subluxation at a different level. We present the details of these five patients and discuss the problems associated with their diagnosis and treatment.


National School Lunch Program and School Breakfast Program: Nutrition Standards for All Foods Sold in School as Required by the Healthy, Hunger-Free Kids Act of 2010. Final rule and interim final rule.
This rule adopts as final, with some modifications, the National School Lunch Program and School Breakfast Program regulations set forth in the interim final rule published in the Federal Register on June 28, 2013. The requirements addressed in this rule conform to the provisions in the Healthy, Hunger-Free Kids Act of 2010 regarding nutrition standards for all foods sold in schools, other than food sold under the lunch and breakfast programs. Most provisions of this final rule were implemented on July 1, 2014, a full year subsequent to publication of the interim final rule. This was in compliance with section 208 of the Healthy, Hunger-Free Kids Act of 2010, which required that State and local educational agencies have at least one full school year from the date of publication of the interim final rule to implement the competitive food provisions. Based on comments received on the interim final rule and implementation experience, this final rule makes a few modifications to the nutrition standards for all foods sold in schools implemented on July 1, 2014. In addition, this final rule codifies specific policy guidance issued after publication of the interim rule. Finally, this rule retains the provision related to the standard for total fat as interim and requests further comment on this single standard.


Poly(N,N-dimethylaminoethyl methacrylate) Brushes: pH-Dependent Switching Kinetics of a Surface-Grafted Thermoresponsive Polyelectrolyte.
The temperature-dependent switching behavior of poly(N,N-dimethylaminoethyl methacrylate) brushes in alkaline, neutral, and acidic solutions is examined. A novel microscopic laser temperature-jump technique is employed in order to study characteristic thermodynamic and kinetic parameters. Static laser micromanipulation experiments allow one to determine the temperature-dependent variation of the swelling ratio. The data reveal a strong shift of the volume phase transition of the polymer brushes to higher temperatures when going from pH = 10 to pH = 4. Dynamic laser micromanipulation experiments offer a temporal resolution on a submillisecond time scale and provide a means to determine the intrinsic rate constants. Both the swelling and the deswelling rates strongly decrease in acidic solutions. Complementary experiments using in situ atomic force microscopy show an increased polymer layer thickness at these conditions. The data are discussed on the basis of pH-dependent structural changes of the polymer brushes including protonation of the amine groups and conformational rearrangements. Generally, repulsive electrostatic interactions and steric effects are assumed to hamper and slow down temperature-induced switching in acidic solutions. This imposes significant restrictions for smart polymer surfaces, sensors, and devices requiring fast response times.


A multiplex real-time PCR assay targeting virulence and resistance genes in Salmonella enterica serotype Typhimurium.
Typhimurium is the main serotype of Salmonella enterica subsp. enterica implicated in food-borne diseases worldwide. This study aimed to detect the prevalence of ten markers combined in a macro-array based on multiplex real-time PCR. We targeted characteristic determinants located on pathogenicity islands (SPI-2 to -5, virulence plasmid pSLT and Salmonella genomic island 1 (SGI1)) as well as a specific 16S-23S rRNA intergenic spacer sequence of definitive type 104 (DT104). To investigate antimicrobial resistance, the study also targeted the presence of genes involved in sulfonamide (sul1) and beta-lactam (blaTEM) resistance. Finally, the intI1 determinant encoding integrase from class 1 integron was also investigated. A total of 538 unrelated S. Typhimurium strains isolated between 1999 and 2009 from various sources, including food animals, food products, human and environmental samples were studied. Based on the combined presence or absence of these markers, we distinguished 34 different genotypes, including three major genotypes encountered in 75% of the studied strains, Although SPI determinants were almost always detected, SGI1, intI1, sul1 and blaTEM determinants were found 47%, 52%, 54% and 12% of the time respectively, varying according to isolation source. Low-marker patterns were most often detected in poultry sources whereas full-marker patterns were observed in pig, cattle and human sources. The GeneDisc® assay developed in this study madeit easier to explore variability within serotype Typhimurium by analyzing ten relevant gene determinants in a large collection of strains. This real-time multiplex method constitutes a valuable tool for strains characterization on epidemiological purposes.


Circulating steroid hormone levels and risk of breast cancer for postmenopausal women.
Epidemiologic studies have consistently reported that endogenous steroid hormone levels are associated with postmenopausal breast cancer risk, but little is known on the associations by tumor grade, hormone receptor status, or age at diagnosis. We performed a case-cohort study of naturally postmenopausal women within the Melbourne Collaborative Cohort Study that included a random sample of 857 women and 197 breast cancer cases diagnosed during a mean of 9.2 years of follow-up. Concentrations of total estradiol, estrone sulfate, testosterone, DHEA sulfate, androstenedione, and sex hormone binding globulin were measured in plasma collected at baseline before diagnosis; free estradiol plasma concentration was calculated. Cox regression was used to estimate associations adjusted for known and potential confounders. The HR for breast cancer comparing fourth and first quartiles was 1.44 [95% confidence interval (95% CI), 0.89-2.35] for total estradiol, 1.75 (95% CI, 1.06, 2.89) for free estradiol, 2.05 (95% CI, 1.24-3.37) for estrone sulfate, 1.25 (95% CI, 0.78-2.01) for testosterone, 1.41 (95% CI, 0.88-2.27) for DHEA sulfate, 1.49 (95% CI, 0.91-2.44) for androstenedione, and 0.33 (95% CI, 0.19-0.55) for sex hormone binding globulin. These associations did not differ by tumor grade and estrogen receptor/progesterone receptor status (all test for heterogeneity, P > 0.05). Risks associated with estrogen and androgen levels were stronger at older ages (test for interaction across age groups, P = 0.59 for total estradiol and P = 0.01 for testosterone). Our prospective study confirms earlier findings and suggests that the associations of endogenous hormones with postmenopausal breast cancer risk are independent of tumor grade, and hormone receptor status and might increase in strength with age.


"An honest tale speeds best, being plainly told": another look at health spending and the supply of physicians.
Claims that the current physician resource shortage is due to the reduced work effort of physicians are misleading and ignore important trends, namely the demographic changes within the profession, the growth in non-physician spending that has outpaced spending on physicians and the relative decline in spending on physician services over the past 20 years. Such data make it difficult to support Evans's and McGrail's (2008) assertions, which distract from more fruitful policy discussions about eliminating the current shortage of physicians, integrating non-physician health providers into medical practice and otherwise meeting the growing demand for health services.


The cost of implementing a jail diversion program for people with mental illness in San Antonio, Texas.
Jail diversion programs for people with mental illness are designed to redirect offenders with mental illness into community treatment. Although much has been published about program models and their successes, little detail is available to policy makers and community stakeholders on the resources required to start and implement a jail diversion program and which agencies bear how much of the burden. The current study used data on a model jail diversion program in San Antonio, Texas, to address this research gap. Data on staff costs, client contacts, planning, and implementation were collected for three types of diversion: pre-booking police, post-booking bond, and post-booking docket. An activity-based costing algorithm was developed to which parameter values were applied. The start-up cost for the program was $556,638.69. Pre-booking diversion cost $370 per person; 90% of costs were incurred by community mental health agencies for short-term monitoring and screening (>80% of activities). Post-booking bond and docket diversion cost $238 and $205 per person, respectively; the majority of costs were incurred by the courts for court decisions. Developing a multiple-intercept jail diversion program requires significant up-front investment. The share of costs varies greatly depending on the type of diversion.


Galloylated catechins as potent inhibitors of hypochlorous acid-induced DNA damage.
Hypochlorous acid (HOCl), a strong oxidant derived from myeloperoxidase in neutrophils and macrophages, can chlorinate DNA bases at the site of inflammation. Because little is known about the protective role of natural antioxidants, such as polyphenols, for the myeloperoxidase-derived DNA damage, we screened the inhibitory effects of various phenolic antioxidants on the chlorination of the 2'-deoxycytidine residue by HOCl in vitro and found that green tea catechins, especially (-)-epicatechin gallate (ECg) and (-)-epigallocatechin gallate (EGCg), significantly inhibited the chlorination. These catechins also reduced nucleoside- and taurine-chloramines, which can induce secondary oxidative damage, into their native forms. Mass spectrometric and nuclear magnetic resonance analyses showed that ECg and EGCg can effectively scavenge HOCl and/or chloramine species resulting in the formation of mono- and dichlorinated ECg and EGCg. Using the HL-60 human leukemia cell line, it was found that ECg could efficiently accumulate in the cells. Immunocytometric analyses using antihalogenated 2'-deoxycytidine antibody showed that pretreatment of cells with ECg inhibited the HOCl-induced immunofluorescence. In addition, the chlorinated ECg derivatives were detected in the HOCl-treated HL-60 cells. These results showed that green tea catechins, especially 3-galloylated catechins, may be the plausible candidate for the prevention of inflammation-derived DNA damage and perhaps carcinogenesis.


Fibroblast growth factors as tissue repair and regeneration therapeutics.
Cell communication is central to the integration of cell function required for the development and homeostasis of multicellular animals. Proteins are an important currency of cell communication, acting locally (auto-, juxta-, or paracrine) or systemically (endocrine). The fibroblast growth factor (FGF) family contributes to the regulation of virtually all aspects of development and organogenesis, and after birth to tissue maintenance, as well as particular aspects of organism physiology. In the West, oncology has been the focus of translation of FGF research, whereas in China and to an extent Japan a major focus has been to use FGFs in repair and regeneration settings. These differences have their roots in research history and aims. The Chinese drive into biotechnology and the delivery of engineered clinical grade FGFs by a major Chinese research group were important enablers in this respect. The Chinese language clinical literature is not widely accessible. To put this into context, we provide the essential molecular and functional background to the FGF communication system covering FGF ligands, the heparan sulfate and Klotho co-receptors and FGF receptor (FGFR) tyrosine kinases. We then summarise a selection of clinical reports that demonstrate the efficacy of engineered recombinant FGF ligands in treating a wide range of conditions that require tissue repair/regeneration. Alongside, the functional reasons why application of exogenous FGF ligands does not lead to cancers are described. Together, this highlights that the FGF ligands represent a major opportunity for clinical translation that has been largely overlooked in the West.


Prognostic factors for advanced-stage human immunodeficiency virus-associated classical Hodgkin lymphoma treated with doxorubicin, bleomycin, vinblastine, and dacarbazine plus combined antiretroviral therapy: a multi-institutional retrospective study.
The treatment and outcomes of patients with human immunodeficiency virus (HIV)-associated Hodgkin lymphoma (HL) continue to evolve. The International Prognostic Score (IPS) is used to predict the survival of patients with advanced-stage HL, but it has not been validated in patients with HIV infection. This was a multi-institutional, retrospective study of 229 patients with HIV-associated, advanced-stage, classical HL who received doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) plus combination antiretroviral therapy. Their clinical characteristics were presented descriptively, and multivariate analyses were performed to identify the factors that were predictive of response and prognostic of progression-free survival (PFS) and overall survival (OS). The overall and complete response rates to ABVD in patients with HIV-associated HL were 91% and 83%, respectively. After a median follow-up of 5 years, the 5-year PFS and OS rates were 69% and 78%, respectively. In multivariate analyses, there was a trend toward an IPS score >3 as an adverse factor for PFS (hazard ratio [HR], 1.49; P=.15) and OS (HR, 1.84; P=.06). A cluster of differentiation 4 (CD4)-positive (T-helper) cell count <200 cells/μL was associated independently with both PFS (HR, 2.60; P=.002) and OS (HR, 2.04; P=.04). The CD4-positive cell count was associated with an increased incidence of death from other causes (HR, 2.64; P=.04) but not with death from HL-related causes (HR, 1.55; P=.32). The current results indicate excellent response and survival rates in patients with HIV-associated, advanced-stage, classical HL who receive ABVD and combination antiretroviral therapy as well as the prognostic value of the CD4-positive cell count at the time of lymphoma diagnosis for PFS and OS.


Safety of cellobiose as a novel food pursuant to regulation (EU) 2015/2283.
Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver an opinion on cellobiose as a novel food (NF) pursuant to Regulation (EU) 2015/2283. The NF consists of two glucose monomers linked by a β-(1-4) glucosidic bond. The applicant intends to add the NF to a variety of foods, and to food supplements aimed at the general population 3 years and older. The information provided on the manufacturing process, composition and specifications of the NF is sufficient and does not raise safety concerns. The applicant provided a subchronic toxicological study which did not raise safety concerns. The applicant provided a human dose-escalation study from which the Panel concludes that the consumption of 20 g per day of cellobiose (equivalent to 290 mg/kg body weight (bw) per day in a 70-kg adult) does not raise concern regarding gastrointestinal tolerability. The maximum anticipated daily intake of cellobiose from the proposed uses is below 290 mg/kg bw per day in the target population. Considering the nature, source, compositional characterisation, and production process of the NF, as well as the toxicological data provided, the Panel considers that the NF does not raise safety concerns under the proposed conditions of use.


An experimental test of the contributions and condition dependence of microstructure and carotenoids in yellow plumage coloration.
A combination of structural and pigmentary components is responsible for many of the colour displays of animals. Despite the ubiquity of this type of coloration, neither the relative contribution of structures and pigments to variation in such colour displays nor the relative effects of extrinsic factors on the structural and pigment-based components of such colour has been determined. Understanding the sources of colour variation is important because structures and pigments may convey different information to conspecifics. In an experiment on captive American goldfinches Carduelis tristis, we manipulated two parameters, carotenoid availability and food availability, known to affect the expression of carotenoid pigments in a full-factorial design. Yellow feathers from these birds were then analysed in two ways. First, we used full-spectrum spectrometry and high-performance liquid chromatography to examine the extent to which variation in white structural colour and total carotenoid content was associated with variation in colour properties of feathers. The carotenoid content of yellow feathers predicted two colour parameters (principal component 1--representing high values of ultraviolet and yellow chroma and low values of violet-blue chroma-and hue). Two different colour parameters (violet-blue and yellow chroma) from white de-pigmented feathers, as well as carotenoid content, predicted reflectance measurements from yellow feathers. Second, we determined the relative effects of our experimental manipulations on white structural colour and yellow colour. Carotenoid availability directly affected yellow colour, while food availability affected it only in combination with carotenoid availability. None of our manipulations had significant effects on the expression of white structural colour. Our results suggest that the contribution of microstructures to variation in the expression of yellow coloration is less than the contribution of carotenoid content, and that carotenoid deposition is more dependent on extrinsic variability than is the production of white structural colour.


Point spread function and two-point resolution in Fresnel incoherent correlation holography.
Fresnel Incoherent Correlation Holography (FINCH) allows digital reconstruction of incoherently illuminated objects from intensity records acquired by a Spatial Light Modulator (SLM). The article presents wave optics model of FINCH, which allows analytical calculation of the Point Spread Function (PSF) for both the optical and digital part of imaging and takes into account Gaussian aperture for a spatial bounding of light waves. The 3D PSF is used to determine diffraction limits of the lateral and longitudinal size of a point image created in the FINCH set-up. Lateral and longitudinal resolution is investigated both theoretically and experimentally using quantitative measures introduced for two-point imaging. Dependence of the resolving power on the system parameters is studied and optimal geometry of the set-up is designed with regard to the best lateral and longitudinal resolution. Theoretical results are confirmed by experiments in which the light emitting diode (LED) is used as a spatially incoherent source to create object holograms using the SLM.


Use of Pathogen-Specific Antibody Biomarkers to Estimate Waterborne Infections in Population-Based Settings.
This review discusses the utility of pathogen-specific antibody biomarkers for improving estimates of the population burden of waterborne infections, assessing the fraction of infections that can be prevented by specific water treatments, and understanding transmission routes and the natural history and ecology of disease in different populations (including asymptomatic infection rates). We review recent literature on the application of pathogen-specific antibody response data to estimate incidence and prevalence of acute infections and their utility to assess the contributions of waterborne transmission pathways. Advantages and technical challenges associated with the use of serum versus minimally invasive salivary antibody biomarkers in cross-sectional and prospective surveys are discussed. We highlight recent advances and challenges and outline future directions for research, development, and application of antibody-based and other immunological biomarkers of waterborne infections.


Factors associated with emergency department presentation after total joint arthroplasty: a population-based retrospective cohort study.
Unplanned visits to the emergency department after total joint arthroplasty are far more common than unplanned readmissions. Our objectives were to characterize the prevalence of presentation to an emergency department for any reason after total joint arthroplasty and to identify risk factors for such visits. Using health administrative databases, we conducted a population-based retrospective cohort study of adults (19-89 yr of age) who received their first primary elective total hip arthroplasty (THA) or total knee arthroplasty (TKA) procedure for arthritis between April 2011 and March 2016 in Ontario. We made univariate comparisons between patients who presented to the emergency department within 30 days of surgery and those who did not in. We determined differences in use of health care services between groups by comparing the change in use in the year before and after surgery between patients who presented to the emergency department and those who did not. We developed logistic regression models for the occurrence of an emergency department visit using backward variable elimination. We identified 42 273 total hip recipients and 70 725 total knee recipients, of whom 5640 (13.3%) and 11 224 (15.9%), respectively, presented to the emergency department within 30 days of surgery. Fewer than 1% of these patients required admission, and nearly half (45%) went to a different institution from where they had their surgery. Among both THA and TKA recipients, patients who presented to the emergency department had a net increase in their median annual health care costs (THA: $501, TKA: $682), compared to a net decrease for the cohort as a whole. Factors associated with increased risk of an emergency visit included increased patient age, male sex, rural residence and various comorbidities. Predictive regression models showed poor discriminative ability for both THA (C-statistic 0.57) and TKA (C-statistic 0.58) recipients. One in 7 patients presented to the emergency department within 30 days of THA or TKA. Some may conceivably have been managed remotely, and very few required readmission. There is a crucial need for strategies to minimize these events.


Lessons learned from the polio eradication initiative in the Democratic Republic of Congo and Ethiopia: analysis of implementation barriers and strategies.
Since its inception in 1988, the Global Polio Eradication Initiative (GPEI) has partnered with 200 countries to vaccinate over 2.5 billion children against poliomyelitis. The polio eradication approach has adapted to emerging challenges and diverse contexts. Knowledge assets gained from these experiences can inform implementation of future health programs, but only if efforts are made to systematically map barriers, identify strategies to overcome them, identify unintended consequences, and compare experiences across country contexts. A sequential explanatory mixed methods design, including an online survey followed by key informant interviews (KIIs), was utilized to map tacit knowledge derived from the polio eradication experience from 1988 to 2019. The survey and KIIs were conducted between September 2018 and March 2019. A cross-case comparison was conducted of two study countries, the Democratic Republic of Congo (DRC) and Ethiopia, which fit similar epidemiological profiles for polio. The variables of interest (implementation barriers, strategies, unintended consequences) were compared for consistencies and inconsistencies within and across the two country cases. Surveys were conducted with 499 and 101 respondents, followed by 23 and 30 KIIs in the DRC and Ethiopia, respectively. Common implementation barriers included accessibility issues caused by political insecurity, population movement, and geography; gaps in human resources, supply chain, finance and governance; and community hesitancy. Strategies for addressing these barriers included adapting service delivery approaches, investing in health systems capacity, establishing mechanisms for planning and accountability, and social mobilization. These investments improved system infrastructure and service delivery; however, resources were often focused on the polio program rather than strengthening routine services, causing community mistrust and limiting sustainability. The polio program investments in the DRC and Ethiopia facilitated program implementation despite environmental, system, and community-level barriers. There were, however, missed opportunities for integration. Remaining pockets of low immunization coverage and gaps in surveillance must be addressed in order to prevent importation of wild poliovirus and minimize circulating vaccine-derived poliovirus. Studying these implementation processes is critical for informing future health programs, including identifying implementation tools, strategies, and principles which can be adopted from polio eradication to ensure health service delivery among hard-to-reach populations. Future disease control or eradication programs should also consider strategies which reduce parallel structures and define a clear transition strategy to limit long-term external dependency.


Increased Turnover at Limiting O2 Concentrations by the Thr(387) → Ala Variant of HIF-Prolyl Hydroxylase PHD2.
PHD2 is a 2-oxoglutarate, non-heme Fe(2+)-dependent oxygenase that senses O2 levels in human cells by hydroxylating two prolyl residues in the oxygen-dependent degradation domain (ODD) of HIF1α. Identifying the active site contacts that determine the rate of reaction at limiting O2 concentrations is crucial for understanding how this enzyme senses pO2 and may suggest methods for chemically altering hypoxia responses. A hydrogen bonding network extends from the Fe(II) cofactor through ordered waters to the Thr(387) residue in the second coordination sphere. Here we tested the impact of the side chain of Thr(387) on the reactivity of PHD2 toward O2 through a combination of point mutagenesis, steady state kinetic experiments and {FeNO}(7) EPR spectroscopy. The steady state kinetic parameters for Thr(387) → Asn were very similar to those of wild-type (WT) PHD2, but kcat and kcat/KM(O2) for Thr(387) → Ala were increased by roughly 15-fold. X-Band electron paramagnetic resonance spectroscopy of the {FeNO}(7) centers of the (Fe+NO+2OG) enzyme forms showed the presence of a more rhombic line shape in Thr(387) → Ala than in WT PHD2, indicating an altered conformation for bound gas in this variant. Here we show that the side chain of residue Thr(387) plays a significant role in determining the rate of turnover by PHD2 at low O2 concentrations.


Tissue transglutaminase 2 inhibition promotes cell death and chemosensitivity in glioblastomas.
Tissue transglutaminase 2 belongs to a family of transglutaminase proteins that confers mechanical resistance from proteolysis and stabilizes proteins. Transglutaminase 2 promotes transamidation between glutamine and lysine residues with the formation of covalent linkages between proteins. Transglutaminase 2 also interacts and forms complexes with proteins important in extracellular matrix organization and cellular adhesion. We have identified the novel finding that treatment of glioblastoma cells with transglutaminase 2 inhibitors promotes cell death and enhances sensitivity to chemotherapy. Treatment with either the competitive transglutaminase 2 inhibitor, monodansylcadaverine, or with highly specific small-molecule transglutaminase 2 inhibitors, KCA075 or KCC009, results in induction of apoptosis in glioblastoma cells. Treatment with these transglutaminase 2 inhibitors resulted in markedly decreased levels of the prosurvival protein, phosphorylated Akt, and its downstream targets. These changes promote a proapoptotic profile with altered levels of multiple intracellular proteins that determine cell survival. These changes include decreased levels of the antiapoptotic proteins, survivin, phosphorylated Bad, and phosphorylated glycogen synthetase kinase 3beta (GSK-3beta), and increased levels of the proapoptotic BH3-only protein, Bim. In vivo studies with s.c. murine DBT glioblastoma tumors treated with transglutaminase 2 inhibitors combined with the chemotherapeutic agent, N-N'-bis (2-chloroethyl)-N-nitrosourea (BCNU), decreased tumor size based on weight by 50% compared with those treated with BCNU alone. Groups treated with transglutaminase 2 inhibitors showed an increased incidence of apoptosis determined with deoxynucleotidyl transferase-mediated biotin nick-end labeling staining. These studies identify inhibition of transglutaminase 2 as a potential target to enhance cell death and chemosensitivity in glioblastomas.


Ethics commentary: subjects of knowledge and control in field primatology.
Our primate kin are routinely displaced from their habitats, hunted for meat, captured for trade, housed in zoos, made to perform for our entertainment, and used as subjects in biomedical testing. They are also the subjects of research inquiries by field primatologists. In this article, we place primate field studies on a continuum of human and alloprimate relationships as a heuristic device to explore the unifying ethical implications of such inter-relationships, as well as address specific ethical challenges arising from common research protocols "in the field" (e.g. risks associated with habituation, disease transmission, invasive collection of biological samples, etc.). Additionally, we question the widespread deployment of conservation- and/or local economic development-based justifications for field-based primatological pursuits. Informed by decades of combined fieldwork experience in Indonesia and the Democratic Republic of Congo, we demonstrate the process by which the adherence to a particular ethical calculus can lead to unregulated and ethically problematic research agendas. In conclusion, we offer several suggestions to consider in the establishment of a formalized code of ethics for field primatology.


Hospital-acquired conditions after bariatric surgery: we can predict, but can we prevent?
Centers for Medicare and Medicaid Services initiated a non-payment policy for certain hospital-acquired conditions (HACs) in 2008. This study aimed to determine the rate of the three most common HACs (surgical site infection (SSI), urinary tract infection (UTI), and venous thromboembolism (VTE)) among bariatric surgery patients. Additionally, the association of HACs with patient factors and the effect of HACs on post-operative outcomes were investigated. Patients over 18 years with a body mass index (BMI) ≥ 35 who underwent bariatric surgery were identified using the American College of Surgeons' National Surgical Quality Improvement Program (ACS-NSQIP) database (2005-2012). Patients were grouped into two categories: HAC versus no HAC patients and baseline characteristics and outcomes, including 30-day mortality, reoperation, and mean length of stay (LOS) were compared. Multivariable logistic regression analysis was performed to identify the risk factors for developing a HAC. 98,553 patients were identified, 2,809 (2.9%) developed at least one HACs. SSI was the most common HAC (1.8%), followed by UTI (0.7%) and VTE (0.4%). The rate of these HACs significantly decreased from 4.6% in 2005-2006 to 2.5% in 2012 (p < 0.001). Laparoscopic gastric banding was associated with the lowest rates of HAC (1.3%) and open gastric bypass with the highest (8.0%). HAC patients had significantly higher rates of in-hospital mortality (0.8 vs. 0.1%, p < 0.001) and LOS (3.9 vs. 2.1 days, p < 0.001). On adjusted analysis, open GBP patients had 5.36-fold higher odds of developing a HAC. Interestingly, the presence of a resident surgeon 7-11 years post graduation was associated with significantly increased odds of HACs (1.86, 1.50-2.31, p < 0.001). Our data demonstrate a strong correlation between these three HACs following bariatric surgery and factors intrinsic to the bariatric patient population. This calls into question the non-payment policy for inherent patient factors on which they cannot have impact. These findings are important to help inform health care policy decisions regarding access to care for bariatric surgery patients.


Factors associated with HIV prevalence in a pre-partum cohort of Zambian women.
An ongoing study of mother-to-child human herpes virus-8 (HHV-8) transmission in Zambian women (n = 3160) allowed us to examine the association of medical injections with HIV serostatus while simultaneously accounting for other factors known to be correlated with HIV prevalence. Multi-method data collection included structured interviews, medical record abstraction, clinical examinations, and biological measures. Medically administered intramuscular or intravenous injections in the past five years (but not blood transfusions) were overwhelmingly correlated with HIV prevalence, exceeding the contribution of sexual behaviours in a multivariable logistic regression. Statistically significant associations with HIV also were found for some demographic variables, sexual behaviours, alcohol use, and sexually transmitted diseases (STD). The results confirmed that iatrogenic needle exposure, sexual behaviour, demographic factors, substance use, and STD history are all implicated in Zambian women's HIV+ status. However, the disproportionate association of medical injection history with HIV highlights the need to investigate further and prospectively the role of health-care injection in sub-Saharan Africa's HIV epidemic.


A pilot study on the feasibility of European harmonized human biomonitoring: Strategies towards a common approach, challenges and opportunities.
In 2004 the European Commission and Member States initiated activities towards a harmonized approach for Human Biomonitoring surveys throughout Europe. The main objective was to sustain environmental health policy by building a coherent and sustainable framework and by increasing the comparability of data across countries. A pilot study to test common guidelines for setting up surveys was considered a key step in this process. Through a bottom-up approach that included all stakeholders, a joint study protocol was elaborated. From September 2011 till February 2012, 17 European countries collected data from 1844 mother-child pairs in the frame of DEMOnstration of a study to COordinate and Perform Human Biomonitoring on a European Scale (DEMOCOPHES).(1) Mercury in hair and urinary cadmium and cotinine were selected as biomarkers of exposure covered by sufficient analytical experience. Phthalate metabolites and Bisphenol A in urine were added to take into account increasing public and political awareness for emerging types of contaminants and to test less advanced markers/markers covered by less analytical experience. Extensive efforts towards chemo-analytical comparability were included. The pilot study showed that common approaches can be found in a context of considerable differences with respect to experience and expertize, socio-cultural background, economic situation and national priorities. It also evidenced that comparable Human Biomonitoring results can be obtained in such context. A European network was built, exchanging information, expertize and experiences, and providing training on all aspects of a survey. A key challenge was finding the right balance between a rigid structure allowing maximal comparability and a flexible approach increasing feasibility and capacity building. Next steps in European harmonization in Human Biomonitoring surveys include the establishment of a joint process for prioritization of substances to cover and biomarkers to develop, linking biomonitoring surveys with health examination surveys and with research, and coping with the diverse implementations of EU regulations and international guidelines with respect to ethics and privacy.


Drivers and barriers to acceptance of human-papillomavirus vaccination among young women: a qualitative and quantitative study.
Human papillomavirus (HPV) is a necessary cause of cervical dysplasia and cancer, and of genital warts. Few studies have examined attitudes to HPV vaccination since the introduction of HPV vaccines. We aimed to investigate the reasons for young women's acceptance or rejection of the quadrivalent HPV vaccine after its general availability in Denmark. A literature review assessed attitudes towards HPV vaccination and the information was used to identify relevant questions for telephone and focus group interviews with women aged 16-26 who had decided to receive or reject HPV vaccination. 435 women across Denmark were interviewed by telephone. Qualitative interviews were undertaken in focus groups with 33 women living in Odense who had completed the telephone survey. Four focus groups were set up according to age (16-20 and 21-26 years of age) and acceptance/rejection of the vaccine. Of 839 women initially contacted by telephone, 794 were included, 411 (49%) said they accepted vaccination but only 201 (24%) had actually received the vaccine and these latter were interviewed. 242 women said they refused vaccination of which 234 were interviewed. Women who were undecided were excluded from the study. Prevention of cervical cancer was the main driver for acceptance of the vaccine, followed by parental encouragement and financial support, personal experience of someone with cancer and recommendation by health-care professionals. The greatest barrier to vaccination was its cost. A lack of information about the benefits of vaccination for sexually active women was also an important barrier and the older participants in particular considered that they were too old to be vaccinated. Knowledge about HPV and its role in the development of cervical cancer and genital warts was poor. The difference between intention to be vaccinated and starting vaccination was considerable, and a large proportion of women aged 16-26 did not wish to be vaccinated. If the most important barriers to vaccination were addressed (cost and a lack of information about vaccination benefits), it is likely that the uptake of vaccination in Denmark would increase substantially.


Clinical efficacy and cost-effectiveness evidence of spa therapy in osteoarthritis. The results of "Naiade" Italian Project.
The "Naiade" Project was carried out in execution of the Decree of December 1994 by the Italian Ministry of Health, with the objective of defining the therapeutic role of spa therapy in the various sectors of medicine. In this study the authors refer to the results obtained in osteoarthritis (OA). The OA study included the observation of patients for 2 consecutive years, with the compilation of a series of clinical and socio-economic data. The initial sample was of 11437 patients suffering from primary and secondary OA, and in the 2(nd) year the total number of assessable patients was 6111 (53.4%). All patients underwent 1 annual cycle of balneotherapy and mud packs therapy for 2 consecutive years. The thermal treatments were carried out in 98 Italian spas with sulphurous water, sodium chloride-bromide-iodide water, sulphate water, and bicarbonate water. Analysis of the data collected confirmed the clinical efficacy of spa treatments for OA, particularly for localisations in the cervical and lumbar spine. The benefits of these treatments and the persistence of the therapeutic effects over time were clearly demonstrated by the collection of some socio-economic indicators relative to the year before each cycle of thermal therapy. In fact, the analysis of these indicators showed a significant reduction in recourse to additional treatments (hospital admissions, physical and pharmacological therapies) and absence from work. The obtained data was particularly interesting for the number of samples examined, the follow-up over a period of 2 years, and the collection of clinical and, especially, socio-economic parameters. The results of the Naiade Project confirmed the value of spa therapies in the treatment of OA.


High resolution melting for mutation scanning of TP53 exons 5-8.
p53 is commonly inactivated by mutations in the DNA-binding domain in a wide range of cancers. As mutant p53 often influences response to therapy, effective and rapid methods to scan for mutations in TP53 are likely to be of clinical value. We therefore evaluated the use of high resolution melting (HRM) as a rapid mutation scanning tool for TP53 in tumour samples. We designed PCR amplicons for HRM mutation scanning of TP53 exons 5 to 8 and tested them with DNA from cell lines hemizygous or homozygous for known mutations. We assessed the sensitivity of each PCR amplicon using dilutions of cell line DNA in normal wild-type DNA. We then performed a blinded assessment on ovarian tumour DNA samples that had been previously sequenced for mutations in TP53 to assess the sensitivity and positive predictive value of the HRM technique. We also performed HRM analysis on breast tumour DNA samples with unknown TP53 mutation status. One cell line mutation was not readily observed when exon 5 was amplified. As exon 5 contained multiple melting domains, we divided the exon into two amplicons for further screening. Sequence changes were also introduced into some of the primers to improve the melting characteristics of the amplicon. Aberrant HRM curves indicative of TP53 mutations were observed for each of the samples in the ovarian tumour DNA panel. Comparison of the HRM results with the sequencing results revealed that each mutation was detected by HRM in the correct exon. For the breast tumour panel, we detected seven aberrant melt profiles by HRM and subsequent sequencing confirmed the presence of these and no other mutations in the predicted exons. HRM is an effective technique for simple and rapid scanning of TP53 mutations that can markedly reduce the amount of sequencing required in mutational studies of TP53.


Quantitative Measurements of the Ciliary Body in Eyes With Acute Primary-Angle Closure.
To compare the structural differences of the ciliary body in patients with acute primary-angle closure (APAC) and normal subjects. Forty-four patients with APAC in one eye and 25 eyes from 25 age-matched normal subjects were consecutively recruited. A-scan ultrasound and ultrasound biomicroscopy (UBM) measurements were performed. Ciliary body parameters including maximum ciliary body thickness (CBTmax), ciliary body thickness at point of the scleral spur (CBT0) and 1000 μm from the scleral spur (CBT1000), anterior placement of ciliary body (APCB), and trabecular-ciliary process angle (TCA), as well as biometric measurements, were measured. Average CBTmax was 0.894 ± 0.114, 0.967 ± 0.110, and 1.053 ± 0.103 mm in eyes with APAC, their fellow eyes, and normal eyes, respectively. Average CBT1000 was 0.616 ± 0.111, 0.631 ± 0.088, and 0.842 ± 0.118 mm, respectively. Average TCA was 48.10 ± 13.26°, 50.60 ± 9.08°, and 87.11 ± 20.71°, respectively. CBTmax and CBT0 were thinner in eyes with APAC compared with their fellow eyes (P = 0.002, P < 0.001). In addition, CBTmax, CBT1000, and TCA were smaller whereas APCB was larger in the fellow eyes of APAC patients compared with normal eyes (P = 0.002, P < 0.001, P < 0.001, P < 0.001). The anterior chamber depth (ACD) was smaller whereas lens thickness (LT) was larger in eyes with APAC compared with their fellow eyes (P < 0.001, P = 0.036). Smaller ACD and axial length and larger LT and lens vault were found in the fellow eyes of APAC patients compared with normal eyes (P < 0.001, P < 0.001, P = 0.015, P = 0.001). Ciliary bodies were thinner and more anteriorly rotated in eyes with APAC as well as in their fellow eyes. Further studies are needed to elucidate the relationship between ciliary body parameters and mechanism of APAC.


Reproductive isolation driven by the combined effects of ecological adaptation and reinforcement.
Recent years have seen a resurgence of interest in the process of speciation but few studies have elucidated the mechanisms either driving or constraining the evolution of reproductive isolation. In theory, the direct effects of reinforcing selection for increased mating discrimination where interbreeding produces hybrid offspring with low fitness and the indirect effects of adaptation to different environments can both promote speciation. Conversely, high levels of homogenizing gene flow can counteract the forces of selection. We demonstrate the opposing effects of reinforcing selection and gene flow in Timema cristinae walking-stick insects. The magnitude of female mating discrimination against males from other populations is greatest when migration rates between populations adapted to alternate host plants are high enough to allow the evolution of reinforcement, but low enough to prevent gene flow from eroding adaptive divergence in mate choice. Moreover, reproductive isolation is strongest under the combined effects of reinforcement and adaptation to alternate host plants. Our findings demonstrate the joint effects of reinforcement, ecological adaptation and gene flow on progress towards speciation in the wild.


Advancing magnetic resonance imaging in Crohn's disease.
Crohn's disease (CD) is a lifelong chronic inflammatory bowel disease associated with diarrhea, abdominal pain, bloody stool and often perianal fistulae. Inflammation in CD involves the entire gastrointestinal tract, especially including the small and large bowels, causing irreversible bowel damage. Frequent imaging examinations are necessary to monitor disease activity and to evaluate response to therapeutic interventions, and, furthermore, to predict recurrence in order to provide appropriate treatment. The suitable imaging modality should be reproducible, well tolerated, safe and free of ionizing radiation. In recent years, imaging used in CD has dramatically changed. Cross-sectional imaging techniques such as computed tomography and magnetic resonance imaging (MRI) are used to investigate not only extraluminal abnormalities, but also intraluminal changes. Recently, new techniques such as MR enteroclysis, enterography, colonography and enterocolonography have been developed. These recent advances enable the use of MRI to assess bowel disorders with high sensitivity, specificity and accuracy. MRI can evaluate simultaneously the bowel surface, bowel wall, abdominal abscesses and perianal lesions, such as perianal fistulae and perianal abscesses, without the problem of overlapping bowel loops. Therefore, MRI has the potential for evaluation of the overall disease activity of CD without radiation exposure. We believe that MRI is a suitable first choice imaging modality in the assessment of CD.


Scanning all-fiber-optic endomicroscopy system for 3D nonlinear optical imaging of biological tissues.
An extremely compact all-fiber-optic scanning endomicroscopy system was developed for two-photon fluorescence (TPF) and second harmonic generation (SHG) imaging of biological samples. A conventional double-clad fiber (DCF) was employed in the endomicroscope for single-mode femtosecond pulse delivery, multimode nonlinear optical signals collection and fast two-dimensional scanning. A single photonic bandgap fiber (PBF) with negative group velocity dispersion at two-photon excitation wavelength (i.e. approximately 810 nm) was used for pulse prechirping in replacement of a bulky grating/lens-based pulse stretcher. The combined use of DCF and PBF in the endomicroscopy system made the endomicroscope basically a plug-and-play unit. The excellent imaging ability of the extremely compact all-fiber-optic nonlinear optical endomicroscopy system was demonstrated by SHG imaging of rat tail tendon and depth-resolved TPF imaging of epithelial tissues stained with acridine orange. The preliminary results suggested the promising potential of this extremely compact all-fiber-optic endomicroscopy system for real-time assessment of both epithelial and stromal structures in luminal organs.