PipMaker--a web server for aligning two genomic DNA sequences.
PipMaker (http://bio.cse.psu.edu) is a World-Wide Web site for comparing two long DNA sequences to identify conserved segments and for producing informative, high-resolution displays of the resulting alignments. One display is a percent identity plot (pip), which shows both the position in one sequence and the degree of similarity for each aligning segment between the two sequences in a compact and easily understandable form. Positions along the horizontal axis can be labeled with features such as exons of genes and repetitive elements, and colors can be used to clarify and enhance the display. The web site also provides a plot of the locations of those segments in both species (similar to a dot plot). PipMaker is appropriate for comparing genomic sequences from any two related species, although the types of information that can be inferred (e.g., protein-coding regions and cis-regulatory elements) depend on the level of conservation and the time and divergence rate since the separation of the species. Gene regulatory elements are often detectable as similar, noncoding sequences in species that diverged as much as 100-300 million years ago, such as humans and mice, Caenorhabditis elegans and C. briggsae, or Escherichia coli and Salmonella spp. PipMaker supports analysis of unfinished or "working draft" sequences by permitting one of the two sequences to be in unoriented and unordered contigs.


Awareness of risk factors among persons at risk for lung cancer, chronic obstructive pulmonary disease and sleep apnea: a Canadian population-based study.
To assess awareness among persons at risk for lung cancer, chronic obstructive pulmonary disease (COPD) and sleep apnea regarding symptoms and risk factors of the disease, and their attitudes regarding the disease and toward those who are affected. A quantitative hybrid telephone and Internet survey of a representative population of Canadian adults at risk for at least one of the three diseases was conducted. To measure the awareness and attitudes of First Nations, Inuit and Métis people to these diseases, a proportionate number were also surveyed.  A total of 3626 individuals were contacted. Of these, 3036 (84%) were eligible to participate. Of those at risk for lung cancer and COPD, 65% and 69%, respectively, were due to tobacco smoke exposure. Among those at risk, 72% believed that they were informed about lung cancer compared with 36% for COPD and 56% for sleep apnea. Most respondents were knowledgeable about the common symptoms of lung cancer, COPD and sleep apnea, but were less aware of the impact lifestyle choices could have on the development of these disorders and the availability of treatment. Most of the participants (77%) believed that smoking was an addiction rather than a habit (19%). There were no significant differences in the awareness of risk factors, symptoms and attitudes toward all three lung diseases between First Nations, Inuit and Métis people and the general population. Canadians are reasonably aware of risk factors and symptoms for lung cancer and sleep apnea. However, there is poor awareness of COPD as a disease entity. There is a lack of appreciation for the impact lifestyle choices and changes can have on lung diseases.


HOMOCITRULLINE AND HOMOARGININE SYNTHESIS FROM LYSINE.
After the injection of L-lysine and uniformly labeled L-lysine-carbon-14 into the rat, labeled homocitrulline and homoarginine are found in the liver and kidney. The ingestion of lysine by seven normal adults results in an increased urinary excretion of homocitrulline and homoarginine. These data suggest the occurrence of an unreported metabolic pathway for lysine in the rat and man.


Involvement of the guanine nucleotide exchange factor Vav3 in central nervous system development and plasticity.
Small GTP-hydrolyzing enzymes (GTPases) of the RhoA family play manifold roles in cell biology and are regulated by upstream guanine nucleotide exchange factors (GEFs). Herein, we focus on the GEFs of the Vav subfamily. Vav1 was originally described as a proto-oncogene of the hematopoietic lineage. The GEFs Vav2 and Vav3 are more broadly expressed in various tissues. In particular, the GEF Vav3 may play important roles in the developing nervous system during the differentiation of neural stem cells into the major lineages, namely neurons, oligodendrocytes and astrocytes. We discuss its putative regulatory roles for progenitor differentiation in the developing retina, polarization of neurons and formation of synapses, migration of oligodendrocyte progenitors and establishment of myelin sheaths. We propose that Vav3 mediates the response of various neural cell types to environmental cues.


Associations of Body Mass Index, Visceral Fat Area, Waist Circumference, and Waist-to-Hip Ratio with Cognitive Function in Western China: Results from WCHAT Study.
This study examined the relationship between cognitive performance and obesity parameters, such as body mass index (BMI), visceral fat area (VFA), waist circumference (WC), and waist-to-hip ratio (WHR) in western China. A cross-sectional study. 3914 participants, aged ≥50 years, were recruited in this study. Anthropometrics measurements, life-style factors, chronic disease comorbidities, and sleep qualities were recorded for each participant. Among the anthropometrics, BMI, WC, and WHR were assessed using standard procedures, while VFA was calculated using bioelectrical impedance analysis. Cognitive performance was estimated using the Short Portable Mental Status Questionnaire (SPMSQ). Finally, relationships between cognitive abilities and BMI, VFA, WC, and WHR were evaluated using univariate and multivariate regression analyses. Cognitive decline (CD) occurred at a rate of 13.29% among the 3914 participants. A strong correlation was observed between cognitive abilities and BMI of male patients aged 50-59 yrs (OR 1.116,95% CI1.002-1.242), in the adjusted model. Alternately, WHR was shown to be significantly related to CD in females aged ≥70 years (OR 0.041, 95% CI0.002-0.671). WC was shown to have a strong association with CD in males (OR 1.023,95% CI1.003-1.024). Lastly, WHR was closely connected to CD in participants with BMI < 25 kg/m2 (OR 0.022,95% CI0.002-0.209). Our findings suggest that a higher middle age BMI is associated with CD, whereas, in the elderly population, a higher WHR is related to improved cognitive performance. Further investigation is warranted to elucidate a relationship between VFA and CD.


Genome-wide association studies to identify rice salt-tolerance markers.
Salinity is an ever increasing menace that affects agriculture worldwide. Crops such as rice are salt sensitive, but its degree of susceptibility varies widely between cultivars pointing to extensive genetic diversity that can be exploited to identify genes and proteins that are relevant in the response of rice to salt stress. We used a diversity panel of 306 rice accessions and collected phenotypic data after short (6 h), medium (7 d) and long (30 d) salinity treatment (50 mm NaCl). A genome-wide association study (GWAS) was subsequently performed, which identified around 1200 candidate genes from many functional categories, but this was treatment period dependent. Further analysis showed the presence of cation transporters and transcription factors with a known role in salinity tolerance and those that hitherto were not known to be involved in salt stress. Localization analysis of single nucleotide polymorphisms (SNPs) showed the presence of several hundred non-synonymous SNPs (nsSNPs) in coding regions and earmarked specific genomic regions with increased numbers of nsSNPs. It points to components of the ubiquitination pathway as important sources of genetic diversity that could underpin phenotypic variation in stress tolerance.


Puniceicoccus vermicola gen. nov., sp. nov., a novel marine bacterium, and description of Puniceicoccaceae fam. nov., Puniceicoccales ord. nov., Opitutaceae fam. nov., Opitutales ord. nov. and Opitutae classis nov. in the phylum 'Verrucomicrobia'.
A Gram-negative, chemoheterotrophic, facultatively anaerobic coccus, designated IMCC1545(T), was isolated from the digestive tract of a marine clamworm, Periserrula leucophryna, inhabiting a tidal flat of the Yellow Sea. Cells of strain IMCC1545(T) are non-motile, dividing by binary fission. The predominant fatty acids are anteiso-C(15 : 0) and C(18 : 0). The respiratory quinone is menaquinone-7 and the DNA G+C content is 52.1 mol%. Phylogenetic analyses based on 16S rRNA gene sequences using three treeing algorithms revealed that the strain formed a novel genus-level lineage within the phylum 'Verrucomicrobia'. The most closely related named organisms to strain IMCC1545(T) are 'Fucophilus fucoidanolyticus' SI-1234 (86.5 % 16S rRNA gene sequence similarity), Alterococcus agarolyticus ADT3(T) (81.8 %) and Opitutus terrae PB90-1(T) (80.3 %), which belong to subdivision 4 of the 'Verrucomicrobia'. Subdivision 4 of the 'Verrucomicrobia' (here named Opitutae classis nov.) was divided into two clades, a clade containing strain IMCC1545(T) and a clade containing Opitutus terrae. From the taxonomic data obtained in this study, it is proposed that the new marine isolate be placed into a novel genus and species named Puniceicoccus vermicola gen. nov., sp. nov. (the type strain of Puniceicoccus vermicola is IMCC1545(T)=KCCM 42343(T)=NBRC 101964(T)) within Puniceicoccaceae fam. nov and Puniceicoccales ord. nov in the class Opitutae. The family Opitutaceae fam. nov. and order Opitutales ord. nov. are also formally proposed.


Tumor plasticity interferes with anti-tumor immunity.
Since tumor cell plasticity was first shown to be crucial in tumor promotion and immune surveillance evasion, it has become an issue of intense investigation. Several mechanisms are associated with the acquisition of tumor cell plasticity and immune evasion, including loss of epithelial phenotype through epithelial-to-mesenchymal transition (EMT). We discuss recent evidence revealing that tumor cell plasticity may lead to the emergence of immunoresistant variants and how the tumor microenvironment evolves to shape this plasticity. We argue that targeting carcinoma cell plasticity represents a novel strategy to better control the emergence of resistant variants and to ensure more effective cancer therapies. In this context, the design of innovative integrative immunotherapy approaches is warranted.


Apollo 16 far-ultraviolet camera/spectrograph: Earth observations.
A far-ultraviolet camera/spectograph experiment was operated on the lunar surface during the Apollo 16 mission. Among the data obtained were images and spectra of the terrestrial atmosphere and geocorona in the wavelength range below 1600 angstroms. These gave the spatial distributions and relative intensities of emissions due to atomic hydrogen, atomic oxygen, molecular nitrogen, and other species-some observed spectrographically for the first time.


Reduced Healthcare Use and Apparent Savings with Passive Home Monitoring Technology: A Pilot Study.
To conduct a cost analysis of ambient assisted living technology, which is promising for improving the ability of individuals and care providers to monitor daily activities and gain better awareness through proactive management of health and safety. Three-arm cohort study. Homes of enrollees of a state-based healthcare plan for older adults. Enrollees dually eligible for Medicare and Medicaid (N = 268). Health and safety passive remote patient monitoring (PRPM) systems were installed in enrollees' homes (the intervention group) with monitoring and proactive intervention of a case manager when deviation from baseline subject behavior was detected. Claims data were collected over 12 months to assess healthcare use and costs in the intervention group and to compare use and costs with those of two control groups: a concurrent group of enrollees who declined the technology and a historical cohort matched on age to the participation group. Although the small sample size precluded cost differences that were statistically significant, the participant group used substantially less custodial care, emergency department (ED) services, inpatient stays, and ED costs than the two control groups. In this pilot study, the PRPM system was associated with apparent healthcare cost savings. Although more cost analyses are warranted, ambient assisted living technologies are a potentially valuable investment for older adult care.


Physicochemical characterization of specific and nonspecific polyaromatic hydrocarbon binders in rat and mouse liver cytosol.
The aryl hydrocarbon hydroxylase inducers 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 3-methylcholanthrene, and benzo(a)pyrene were all shown to bind in a saturable manner to a distinct component in cytosol from both rat and C57BL/6J mouse liver. This component was analyzed by gel permeation chromatography on Sephacryl S-300 and by sucrose density gradient centrifugation and was found to have a Stokes radius of 61 +/- 1.5 A and a sedimentation coefficient of 4.4 S under high salt conditions. Based on these parameters, which were identical for the rat and mouse receptor, a molecular weight of 111,000 was calculated. The same Stokes radius and sedimentation coefficient were observed regardless of the ligand used for labeling of the receptor protein [(3H]TCDD, 3-[3H]methylcholanthrene or [3H]benzo(a)pyrene). On the other hand, 3-[3H]methylcholanthrene and [3H]benzo(a)-pyrene exhibited much more nonspecific binding than [3H]TCDD, at least partially due to contaminating serum components, and it cannot be excluded that some previous findings on low molecular weight hepatic "receptors" for these polyaromatic hydrocarbons may actually be explained in this way. Clearly, use of thoroughly perfused livers would seem to be a prerequisite for investigations on specific binding of aryl hydrocarbon hydroxylase inducers to liver cytosol. The rat hepatic TCDD receptor was also shown to be retained on DEAE-Sepharose (eluted at 0.2-0.3 M NaCl), hydroxylapatite (eluted at 0.15-0.17 M phosphate), and heparin-Sepharose (eluted at 0.3-0.4 M NaCl). In conclusion, the TCDD receptor showed similar physicochemical and chromatographic characteristics to those previously reported for the androgen and glucocorticoid receptors. However, ligand competition experiments indicated that the TCDD receptor is not identical to any steroid receptor. In line with this, monoclonal anti-glucocorticoid receptor-IgG antibodies did not react with the TCDD receptor.


3D-printed hierarchical scaffold for localized isoniazid/rifampin drug delivery and osteoarticular tuberculosis therapy.
After surgical treatment of osteoarticular tuberculosis (TB), it is necessary to fill the surgical defect with an implant, which combines the merits of osseous regeneration and local multi-drug therapy so as to avoid drug resistance and side effects. In this study, a 3D-printed macro/meso-porous composite scaffold is fabricated. High dosages of isoniazid (INH)/rifampin (RFP) anti-TB drugs are loaded into chemically modified mesoporous bioactive ceramics in advance, which are then bound with poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) through a 3D printing procedure. The composite scaffolds show greatly prolonged drug release time compared to commercial calcium phosphate scaffolds either in vitro or in vivo. In addition, the drug concentrations on the periphery tissues of defect are maintained above INH/RFP minimal inhibitory concentrations even up to 12 weeks post-surgery, while they are extremely low in blood. Examinations of certain serum enzymes suggest no harm to hepatic or renal functions. Micro-CT evaluations and histology results also indicate partly degradation of the composite scaffolds and new bone growth in the cavity. These results suggest promising applications of our hierarchical composite scaffold in bone regeneration and local anti-TB therapy after osteoarticular TB debridement surgery.


Impact of Induced Moods, Sensation Seeking, and Emotional Contagion on Economic Decisions Under Risk.
In addition to probabilities of monetary gains and losses, personality traits, socio-economic factors, and specific contexts such as emotions and framing influence financial risk taking. Here, we investigated the effects of joyful, neutral, and sad mood states on participants' risk-taking behaviour in a simple task with safe and risky options. We also analysed the effect of framing on risk taking. In different trials, a safe option was framed in terms of either financial gains or losses. Moreover, we investigated the effects of emotional contagion and sensation-seeking personality traits on risk taking in this task. We did not observe a significant effect of induced moods on risk taking. Sad mood resulted in a slight non-significant trend of risk aversion compared to a neutral mood. Our results partially replicate previous findings regarding the presence of the framing effect. As a novel finding, we observed that participants with a low emotional contagion score demonstrated increased risk aversion during a sad mood and a similar trend at the edge of significance was present in high sensation seekers. Overall, our results highlight the importance of taking into account personality traits of experimental participants in financial risk-taking studies.


Assessment of health systems guidance using the Appraisal of Guidelines for Research and Evaluation - Health Systems (AGREE-HS) instrument.
Health systems guidance (HSG) documents contain systematically developed statements or recommendations intended to address a health system challenge. The concept of HSG is fairly new and considerable effort has been undertaken to build tools to support the contextualization of recommendations. One example is the Appraisal of Guidelines for REsearch and Evaluation - Health Systems (AGREE-HS), created by international stakeholders and researchers, to assist in the development, reporting and evaluation of HSG. Here, we present the quality appraisal of 85 HSG documents published from 2012 to 2017 using the AGREE-HS. The AGREE-HS consists of five items (Topic, Participants, Methods, Recommendations, and Implementability), which are scored on a 7-point response scale (1=lowest quality; 7=highest quality). Overall, AGREE-HS item scores were highest for the 'Topic' and 'Recommendations' items (means above the mid-point of 4), while the 'Participants', 'Methods', and 'Implementability' items received lower scores. Documents without a specific health focus and those authored by the National Institute for Health and Care Excellence group, achieved higher AGREE-HS overall scores than their comparators. No statistically significant changes in overall scores were observed over time. This is the first time that the AGREE-HS has been applied, providing a current quality status report of HSG and identifying where improvements in HSG development and reporting can be made.


Erythromycin, retapamulin, pyridoxine, folic acid, and ivermectin inhibit cytopathic effect, papain-like protease, and MPRO enzymes of SARS-CoV-2.
Although tremendous success has been achieved in the development and deployment of effective COVID-19 vaccines, developing effective therapeutics for the treatment of those who do come down with the disease has been with limited success. To repurpose existing drugs for COVID-19, we previously showed, qualitatively, that erythromycin, retapamulin, pyridoxine, folic acid, and ivermectin inhibit SARS-COV-2-induced cytopathic effect (CPE) in Vero cells. This study aimed to quantitatively explore the inhibition of SARS-CoV-2-induced CPE by erythromycin, retapamulin, pyridoxine, folic acid, and ivermectin and to determine the effect of these drugs on SARS-CoV-2 papain-like protease and 3CL protease (MPRO) enzymes. Neutral red (3-amino-7-dimethylamino-2-methyl-phenazine hydrochloride) cell viability assay was used to quantify CPE after infecting pre-treated Vero cells with clinical SARS-Cov-2 isolates. Furthermore, SensoLyte® 520 SARS-CoV-2 papain-like protease and SensoLyte® 520 SARS-CoV-2 MPRO activity assay kits were used to evaluate the inhibitory activity of the drugs on the respective enzymes. Erythromycin, retapamulin, pyridoxine, folic acid, and ivermectin dose-dependently inhibit SARS-CoV-2-induced CPE in Vero cells, with inhibitory concentration-50 (IC50) values of 3.27 µM, 4.23 µM, 9.29 µM, 3.19 µM, and 84.31 µM, respectively. Furthermore, erythromycin, retapamulin, pyridoxine, folic acid, and ivermectin dose-dependently inhibited SARS-CoV-2 papain-like protease with IC50 values of 0.94 µM, 0.88 µM, 1.14 µM, 1.07 µM, and 1.51 µM, respectively, and inhibited the main protease (MPRO) with IC50 values of 1.35 µM, 1.25 µM, 7.36 µM, 1.15 µM, and 2.44 µM, respectively. The IC50 for all the drugs, except ivermectin, was at the clinically achievable plasma concentration in humans, which supports a possible role for the drugs in the management of COVID-19. The lack of inhibition of CPE by ivermectin at clinical concentrations could be part of the explanation for its lack of effectiveness in clinical trials.


Induction of mucous cell metaplasia in the middle ear of rats using a three-step method: an improved model for otitis media with mucoid effusion.
Otitis media with mucoid effusion, characterized by mucous cell metaplasia in the middle ear cleft and thick fluid accumulation in the middle ear cavity, is a common otological disease that frequently affects young children. Multiple factors are involved in the development of this disease, especially middle ear infection and Eustachian tube dysfunction. In this study, in order to induce otitis media with effusion in rats, we introduced a three-step method, namely inoculation of Streptococcus pneumoniae at 10(7) colony-forming units (CFU)/ear or Haemophilus influenzae at 5 x 10(7) CFU/ear into the middle ear cavity twice at 2-week intervals, followed by Eustachian tube obstruction (ETO) for 4 and 8 weeks. Animals inoculated with phosphate-buffered saline (PBS) twice in the same manner followed by ETO served as controls. Middle ear effusion and mucosa were harvested for evaluation of carbohydrate concentrations and mucous cell density, respectively. We found that rats inoculated with S. pneumoniae twice, followed by ETO at 8 weeks, yielded the highest carbohydrate concentration in middle ear effusion and the highest goblet cell density in the middle ear cavity compared to the H. influenzae and PBS groups. It is tentatively concluded that inoculation of S. pneumoniae at 10(7) CFU/ear into the middle ear cavity of rats twice at 2-week intervals, followed by ETO for 8 weeks, is a promising animal model for otitis media with mucoid effusion which may be valuable for studying the human counterpart.


Fluorescence photobleaching analysis of nuclear transport: dynamic evidence for auxiliary channels in detergent-treated nuclei.
Nuclear transport experiments were performed on isolated rat liver nuclei to examine the permeability of membrane and detergent-free peripheral nuclear lamina. The transport of 64K molecular weight fluorescent-derivatized dextrans was measured by using the technique of fluorescence redistribution after photobleaching. Results of these experiments provide evidence for transport pathways that appear to be functionally distinct from nuclear pore complex channels. The suggestion is made that these supplemental pathways are embedded in the peripheral nuclear lamina and are normally masked by the inner nuclear membrane.


Endoscopic retrograde cholangiopancreatography versus intraoperative cholangiography for diagnosis of common bile duct stones.
Endoscopic retrograde cholangiopancreatography (ERCP) and intraoperative cholangiography (IOC) are tests used in the diagnosis of common bile duct stones in people suspected of having common bile duct stones. There has been no systematic review of the diagnostic accuracy of ERCP and IOC. To determine and compare the accuracy of ERCP and IOC for the diagnosis of common bile duct stones. We searched MEDLINE, EMBASE, Science Citation Index Expanded, BIOSIS, and Clinicaltrials.gov to September 2012. To identify additional studies, we searched the references of included studies and systematic reviews identified from various databases (Database of Abstracts of Reviews of Effects (DARE)), Health Technology Assessment (HTA), Medion, and ARIF (Aggressive Research Intelligence Facility)). We did not restrict studies based on language or publication status, or whether data were collected prospectively or retrospectively. We included studies that provided the number of true positives, false positives, false negatives, and true negatives for ERCP or IOC. We only accepted studies that confirmed the presence of common bile duct stones by extraction of the stones (irrespective of whether this was done by surgical or endoscopic methods) for a positive test, and absence of common bile duct stones by surgical or endoscopic negative exploration of the common bile duct, or symptom-free follow-up for at least six months for a negative test as the reference standard in people suspected of having common bile duct stones. We included participants with or without prior diagnosis of cholelithiasis; with or without symptoms and complications of common bile duct stones; with or without prior treatment for common bile duct stones; and before or after cholecystectomy. At least two authors screened abstracts and selected studies for inclusion independently. Two authors independently collected data from each study. We used the bivariate model to summarise the sensitivity and specificity of the tests. We identified five studies including 318 participants (180 participants with and 138 participants without common bile duct stones) that reported the diagnostic accuracy of ERCP and five studies including 654 participants (125 participants with and 529 participants without common bile duct stones) that reported the diagnostic accuracy of IOC. Most studies included people with symptoms (participants with jaundice or pancreatitis) suspected of having common bile duct stones based on blood tests, ultrasound, or both, prior to the performance of ERCP or IOC. Most studies included participants who had not previously undergone removal of the gallbladder (cholecystectomy). None of the included studies was of high methodological quality as evaluated by the QUADAS-2 tool (quality assessment tool for diagnostic accuracy studies). The sensitivities of ERCP ranged between 0.67 and 0.94 and the specificities ranged between 0.92 and 1.00. For ERCP, the summary sensitivity was 0.83 (95% confidence interval (CI) 0.72 to 0.90) and specificity was 0.99 (95% CI 0.94 to 1.00). The sensitivities of IOC ranged between 0.75 and 1.00 and the specificities ranged between 0.96 and 1.00. For IOC, the summary sensitivity was 0.99 (95% CI 0.83 to 1.00) and specificity was 0.99 (95% CI 0.95 to 1.00). For ERCP, at the median pre-test probability of common bile duct stones of 0.35 estimated from the included studies (i.e., 35% of people suspected of having common bile duct stones were confirmed to have gallstones by the reference standard), the post-test probabilities associated with positive test results was 0.97 (95% CI 0.88 to 0.99) and negative test results was 0.09 (95% CI 0.05 to 0.14). For IOC, at the median pre-test probability of common bile duct stones of 0.35, the post-test probabilities associated with positive test results was 0.98 (95% CI 0.85 to 1.00) and negative test results was 0.01 (95% CI 0.00 to 0.10). There was weak evidence of a difference in sensitivity (P value = 0.05) with IOC showing higher sensitivity than ERCP. There was no evidence of a difference in specificity (P value = 0.7) with both tests having similar specificity. Although the sensitivity of IOC appeared to be better than that of ERCP, this finding may be unreliable because none of the studies compared both tests in the same study populations and most of the studies were methodologically flawed. It appears that both tests were fairly accurate in guiding further invasive treatment as most people diagnosed with common bile duct stones by these tests had common bile duct stones. Some people may have common bile duct stones in spite of having a negative ERCP or IOC result. Such people may have to be re-tested if the clinical suspicion of common bile duct stones is very high because of their symptoms or persistently abnormal liver function tests. However, the results should be interpreted with caution given the limited quantity and quality of the evidence.


Medicare savings programs: analyzing options for expanding eligibility.
The Medicare Savings Programs (MSPs) are designed to provide financial assistance to Medicare beneficiaries who do not qualify for full Medicaid coverage. This paper considers changes in eligibility that would better align MSP program rules with those related to receiving low-income subsidies for the Medicare Part D drug benefit. These changes would make more people eligible for the MSPs and could encourage greater participation; similar changes were incorporated in recently passed legislation. Our analysis, based on 2006 data from the Health and Retirement Study, shows there is a trade-off between making larger numbers of beneficiaries eligible by eliminating resource requirements and better targeting of individuals with greater health care needs by expanding income standards.


[Renewed 2015 Clinical Practice Guidelines for Management of Hepatitis C by Korean Association for the Study of the Liver; What Has Been Changed? - Indications for Treatment].
The landscape of treatment for HCV infection has evolved substantially with the advent of highly effective direct-acting antiviral agents (DAA). The Korean Association for the Study of the Liver updated guideline for managemnt of hepatitis C in accordance with the introduction of DAA into practice in late 2015. Due to high effectiveness and few side effects of DAA, indications for treatment has been widened to include patients who had been contraindicated for the combination treatment of peginterferon-α and ribavirin, i.e. decompensated cirrhosis and pre- and post-liver transplant setting. As succeesul treatment of HCV can reduce complications of cirrhosis, development of hepatocelluar carcinoma and liver-related mortality, and improve extrahepatic manifestions, all HCV-infected patients with no contraindication should be considered for treatment. Considering the risk for morbidity and mortality and benefit of treatment, patients with advanced fibrosis ≥F3 including compensated and decompensated cirrhosis, those in the pre- and post-tranplasnt setting, and those with severe extrahepatic manifestations including HCV-related mixed cryoglobulinemia and glomerulonephritis should be given priority for treatment.


Epidemiology, Clinical Features, and Outcome in a Cohort of Adolescents With Cerebral Venous Thrombosis.
We aimed to analyze the epidemiologic, clinical, and paraclinical features of adolescents with cerebral venous thrombosis (CVT) and its therapeutic management and outcome. This multicenter retrospective cohort included patients 10 to 18 years of age hospitalized for a first episode of CVT in 2 French regions between 1999 and 2019. The number of cases was compared to the number recorded by the French health insurance system. The CVT registry of the Lariboisière hospital allowed comparisons with adults. One hundred two patients were included (52.9% female; median age 15.1 years). Estimated incidence was 0.37 to 0.38 per 100,000 adolescents per year; 45.5% of patients presented with focal deficits or seizures or in a coma. Male patients were younger than female patients (14.2 vs 15.6 years; p < 0.01) and more often admitted to intensive care (52.1% vs 24.1%; p = 0.0,035). The lateral sinus was the most common CVT location (72.3%), and 29.4% of adolescents had associated venous infarction or hematoma. Most patients (94.1%) received anticoagulation. Treatment also included an endovascular procedure (2.9%), decompressive craniectomy (4.9%), and CSF shunt (6.9%). The most frequently identified CVT-associated condition was local infection in male (18.6%) and systemic disease in female (14.8%; p < 0.001) patients. The proportion of CVTs in adolescents without an identified associated condition or risk factor was low (1.9% vs 11.4% in adults; p < 0.002). Adverse outcome at 1 year was more frequent than in adults (33.3% vs 11.8%; p = 0.0,001). CVT in adolescents is rare and complex with specific epidemiology, including differences in clinical presentation and associated conditions between sexes, and more severe outcomes than in adults. Careful specialized management and follow-up are therefore recommended.


Hierarchical Data-Driven Analysis of Clinical Symptoms Among Patients With Parkinson's Disease.
Mutations in the LRRK2 and GBA genes are the most common inherited causes of Parkinson's disease (PD). Studies exploring phenotypic differences based on genetic status used hypothesis-driven data-gathering and statistical-analyses focusing on specific symptoms, which may influence the validity of the results. We aimed to explore phenotypic expression in idiopathic PD (iPD) patients, G2019S-LRRK2-PD, and GBA-PD using a data-driven approach, allowing screening of large numbers of features while controlling selection bias. Data was collected from 1525 Ashkenazi Jews diagnosed with PD from the Tel-Aviv Medical center; 161 G2019S-LRRK2-PD, 222 GBA-PD, and 1142 iPD (no G2019S-LRRK2 or any of the 7 AJ GBA mutations tested). Data included 771 measures: demographics, cognitive, physical and neurological functions, performance-based measures, and non-motor symptoms. The association of the genotypes with each of the measures was tested while accounting for age at motor symptoms onset, gender, and disease duration; p-values were reported and corrected in a hierarchical approach for an average over the selected measures false discovery rate control, resulting in 32 measures. GBA-PD presented with more severe symptoms expression while LRRK2-PD had more benign symptoms compared to iPD. GBA-PD presented greater cognitive and autonomic involvement, more frequent hyposmia and REM sleep behavior symptoms while these were less frequent among LRRK2-PD compared to iPD. Using a data-driven analytical approach strengthens earlier studies and extends them to portray a possible unique disease phenotype based on genotype among AJ PD. Such findings could help direct a more personalized therapeutic approach.


Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus.
There is a global need to elucidate protective antigens expressed by the SARS-coronavirus (SARS-CoV). Monoclonal antibody reagents that recognise specific antigens on SARS-CoV are needed urgently. In this report, the development and immunochemical characterisation of a panel of murine monoclonal antibodies (mAbs) against the SARS-CoV is presented, based upon their specificity, binding requirements, and biological activity. Initial screening by ELISA, using highly purified virus as the coating antigen, resulted in the selection of 103 mAbs to the SARS virus. Subsequent screening steps reduced this panel to seventeen IgG mAbs. A single mAb, F26G15, is specific for the nucleoprotein as seen in Western immunoblot while five other mAbs react with the Spike protein. Two of these Spike-specific mAbs demonstrate the ability to neutralise SARS-CoV in vitro while another four Western immunoblot-negative mAbs also neutralise the virus. The utility of these mAbs for diagnostic development is demonstrated. Antibody from convalescent SARS patients, but not normal human serum, is also shown to specifically compete off binding of mAbs to whole SARS-CoV. These studies highlight the importance of using standardised assays and reagents. These mAbs will be useful for the development of diagnostic tests, studies of SARS-CoV pathogenesis and vaccine development.


Phytoestrogens are partial estrogen agonists in the adult male mouse.
The intake, as well as serum and urinary concentrations, of phytoestrogens is high in countries where incidence of prostate cancer is low, suggesting a chemopreventive role for phytoestrogens. Their significance could be explained by the ability to antagonize the action of more potent endogenous estrogens in initiation or promotion of tumor formation. We have studied estrogenicity and antiestrogenicity of dietary soy and two phytoestrogens, coumestrol and daidzein, in our neoDES mouse model for the study or prostatic neoplasia. Soy was chosen because it is rich in phytoestrogens, is widely used in Oriental diets, and has antiestrogenic and anticarcinogenic properties in the neoDES mouse when given from fertilization onward. In short-term tests with adult animals, no evidence for estrogenicity or antiestrogenicity (capability to antagonize the action of 17 beta-estradiol) of soy was found when development of epithelial metaplasia and expression of c-fos protooncogene in prostate were used as end points of estrogen action. Estrogenic activity of coumestrol and daidzein on c-fos expression was subtle. Coumestrol, either given alone or in combination with 17 beta-estradiol, had no effect on development of epithelial metaplasia. These marginal or missing effects in adult males could be interpreted by assuming that the neonatal period is more critical for estrogenic or antiestrogenic action of soy and phytoestrogens. Once initiated, estrogen-related lesions would develop spontaneously. Alternatively, the chemopreventive action of soy is not due to antiestrogenicity of soy-derived phytoestrogens.


Real World Outcomes in Patients With Metastatic, Castration-Resistant Prostate Cancer Treated With Radium-223 in Routine Clinical Practice in Sweden.
Estimate the effect of Radium-223 (Ra-223) on the incidence of bone fractures, prostate cancer death, and all-cause death compared with other standard treatments for metastatic, castration-resistant prostate cancer (mCRPC). Using a cohort design, we estimated the effect of Ra-223 on the risk of bone fractures, all-cause and prostate cancer-specific mortality across different lines of treatment for mCRPC using Prostate Cancer data Base Sweden (2013-2018). The comparator group comprised other standard treatments for mCRPC. We used 36-month risk differences and hazard ratios (HRs) as effect estimates. The number of eligible individuals was 635, 453, 262, and 84 for the first-, second-, third-, and fourth-line cohorts, respectively. When compared Ra-223 to other standard treatments, the difference in the 36-month risk of fracture was 6% (95% confidence interval [CI], -7% to 18%) in the first-line cohort (n = 635) and 8% (95% CI, -7% to 18%) in the second-line cohort (n = 453). The number of fractures in the third-/fourth-line cohorts was too small for an adjusted comparison. The difference in 36-month mortality was higher in the first-line cohort 13% (95% CI, -3% to 31%), but lower in the second- and third-/fourth-line cohorts-8% (95% CI, -23% to 7%) and -14% (95% CI, -21% to 16%) respectively. Most deaths were due to prostate cancer. Results suggest that the difference in the risk of fractures is small, if any. A difference in the risk of mortality may be present in first-line treatment, but a decreased risk of mortality was observed in second and later lines of treatment. The results on mortality need to be considered in the context of potential unmeasured or residual confounding.


Segmental Maternal UPD of Chromosome 7q in a Patient With Pendred and Silver Russell Syndromes-Like Features.
Pendred syndrome (PS) is an autosomal recessive disorder due to mutations in the SLC26A4 gene (chr7q22. 3) and characterized by sensorineural hearing loss and variable thyroid phenotype. Silver-Russell syndrome (SRS) is a heterogeneous imprinting disorder including severe intrauterine and postnatal growth retardation, and dysmorphic features. Maternal uniparental disomy of either the whole chromosome 7 (upd(7)mat) or 7q (upd(7q)mat) is one of the multiple mechanisms impacting the expression of imprinted genes in SRS, and is associated with milder clinical features. Here, we report genetic and clinical characterization of a female child with PS, postnatal growth retardation, and minor dysmorphic features. A gross homozygous deletion of SLC26A4 exons 17-20 was suspected by Sanger sequencing and then confirmed by array-CGH. Moreover, an insertion of about 1 kb of the CCDC126 gene (7p15.3), which does not appear to be clinically relevant, was detected. The possible occurrence of a balanced rearrangement between 7p and 7q was excluded. The absence of the deletion in the father led to the investigation of upd, and microsatellite segregation analysis revealed a segmental 7q (upd(7q)mat), leading to SLC26A4 homozygosity and responsible for both PS and SRS-like traits. The proband matched 3 out of 6 major SRS criteria. In conclusion, this is the first report of uniparental isodisomy encompassing almost the whole long arm of chromosome 7 resulting in PS and SRS-like features. Whereas, the inner ear phenotype of PS is typical, the clinical features suggestive of SRS might have been overlooked.


Electrochemical Oxidation of Hexafluoropropylene Oxide Dimer Acid (GenX): Mechanistic Insights and Efficient Treatment Train with Nanofiltration.
Hexafluoropropylene oxide dimer acid (HFPO-DA, trade name GenX) is a perfluoroalkyl ether carboxylic acid (PFECA) that has been detected in watersheds around the world. Similar to other per- and polyfluoroalkyl substances (PFASs), few processes are able to break HFPO-DA's persistent carbon-fluorine bonds. This study provides both experimental and computational lines of evidence for HFPO-DA mineralization during electrochemical oxidation at a boron-doped diamond anode with a low potential for the generation of stable organofluorine intermediates. Our density functional theory calculations consider the major operative mechanism, direct electron transfer, throughout the entire pathway. Initial oxidative attack does not break the ether bond, but leads to stepwise mineralization of the acidic side chain. Our mechanistic investigations reveal that hydroxyl radicals are unreactive toward HFPO-DA, while electrochemically activated sulfate facilitates its oxidation. Furthermore, we demonstrate that an NF90 membrane is capable of removing 99.5% of HFPO-DA from contaminated water. Electrochemical treatment of the nanofiltration rejectate is shown to reduce both energy and electrode costs by more than 1 order of magnitude compared to direct electrochemical treatment of the raw water. Overall, a nanofiltration-electrochemical oxidation treatment train is a sustainable destructive approach for the cost-effective elimination of HFPO-DA and other PFASs from contaminated water.


The impact of horse purslane (Trianthema portulacastrum L.) infestation on soybean [Glycine max (L.) Merrill] productivity in northern irrigated plains of Pakistan.
Horse purslane (Trianthema portulacstrum L.) is an important weed of soybean crop capable of causing significant yield reduction. Therefore, this study assessed the impact of horse purslane and other weeds' infestation on the productivity of soybean. Ten treatments, i.e., weed-free throughout the growing season, horse purslane-free till 20, 40 and 60 days after emergence (DAE), all weeds-free till 20, 40 and 60 DAE, weedy-check (excluding horse purslane), weedy-check (horse purslane alone) and weedy-check (all weeds) were included in the study. Data relating to density and dry weight of recorded weed species, and yield and related traits of soybean were recorded. Overall, infestation percentage of horse purslane was 33.10 and 51%, whereas dry weight was 12 and 44 g m-2 during 1st and 2nd year, respectively. The highest dry weight of all weed species was recorded at 45 DAE in weedy-check all weeds treatment during both years. The lowest relative density and frequency of horse purslane were recorded in the treatment where it was controlled until 20 DAE during 2018 at 30 DAE, whereas the same treatment recoded the lowest density of horse purslane at 45 DAE during 2019. The relative frequency of horse purslane was non-significant for weedy-check horse purslane and weedy-check all weeds treatments during 2018, whereas former treatment had higher relative frequency of horse purslane in weedy-check all weeds than the later during 2019. Yield and related traits significantly differed among different treatments used in the study. The treatment all weeds controlled until 40 DAE recorded higher number of pods per plant, 1000-seed weight and seed yield during both years. The yield reduction in weedy-check treatments was; weedy-check all weeds > weedy-check all weeds except horse purslane > weedy-check horse purslane only. It is concluded that horse purslane was not the sole weed interfering soybean fields and weed flora consisted of false amaranth [Digera muricata (L.) Mart.] and purple nut sedge (Cyperus rotundus L.). Hence, if the soybean fields in northern irrigated plains of Pakistan are infested with horse purslane or heavily infested with horse purslane or other weeds, these should be controlled in initial 40 DAE to improve soybean productivity.


A tobacco plasma membrane calmodulin-binding transporter confers Ni2+ tolerance and Pb2+ hypersensitivity in transgenic plants.
All organisms require a minimal amount of metal ions to sustain their metabolism, growth and development. At the same time, their intrinsic metal-uptake systems render them vulnerable to toxic levels of metals in the biosphere. Using radiolabeled recombinant calmodulin as a probe to screen a tobacco cDNA library, we have discovered a protein designated NtCBP4 (Nicotiana tabacum calmodulin-binding protein) that can modulate plant tolerance to heavy metals. Structurally, NtCBP4 is similar to vertebrate and invertebrate K+ and to non-selective cation channels, as well as to recently reported proteins from barley and Arabidopsis. Here we report on the subcellular localization of NtCBP4 and the phenotype of transgenic plants overexpressing this protein. The localization of NtCBP4 in the plasma membrane was manifested by fractionating tobacco membranes on sucrose gradients or by aqueous two-phase partitioning, and subsequently using immunodetection. Several independent transgenic lines expressing NtCBP4 had higher than normal levels of NtCBP4. These transgenic lines were indistinguishable from wild type under normal growth conditions. However, they exhibited improved tolerance to Ni2+ and hypersensitivity to Pb2+, which are associated with reduced Ni2+ accumulation and enhanced Pb2+ accumulation, respectively. To our knowledge this is the first report of a plant protein that modulates plant tolerance or accumulation of Pb2+. We propose that NtCBP4 is involved in metal uptake across the plant plasma membrane. This gene may prove useful for implementing selective ion tolerance in crops and improving phytoremediation strategies.


Molecular weight of gas-vesicle protein from the planktonic cyanobacterium Anabaena flos-aquae and implications for structure of the vesicle.
The gas vesicle of the planktonic cyanobacterium Anabaena flos-aquae is a cylindrical shell made of protein enclosing a gas-filled space. Protein sequence analysis shows that the vesicle is made from a single protein. By gel electrophoresis and amino acid analysis its molecular weight was estimated to be 20 600. Taken with previously obtained X-ray data, a simple interpretation of its molecular structure is of the polypeptide snaking in six pairs of antiparallel chains, three in each layer. The molecule would repeat along the ribs of the vesicle at intervals of 3.4 nm.


Multidetector CT Findings of Acquired Spondylolysis and Spondylolisthesis after Posterior Lumbar Laminectomy.
We aimed to analyze postoperative multidetector CT (MDCT) of acquired spondylolysis and spondylolisthesis after posterior lumbar laminectomy. We enrolled 74 patients, from 2003 to 2017, who underwent posterior lumbar laminectomy with both pre and postoperative MDCT. The patients were categorized into the following two groups: group 1 without fusion and group 2 with fusion. We analyzed laminectomy width, level and location of spondylolysis or spondylolisthesis, facet changes, and fatty infiltration of paraspinal muscles on postoperative MDCT. Incidence of spondylolysis or spondylolisthesis was 4 of 20 patients in group 1 and 2 of 54 patients in group 2. The laminectomy width (%) was defined as the percentage of the width of laminectomy to total lamina length. Mean laminectomy width (%) in patients with spondylolysis or spondylolisthesis was 54.0 in group 1 and 53.2 in group 2, in contrast to that in patients without spondylolysis or spondylolisthesis, which was 35.0 in group 1. The spondylolysis was observed at the level of the laminectomy and below pars interarticularis in group 1 and below the fusion mass at isthmic region in group 2. MDCT facilitates the diagnosis of postsurgical acquired spondylolysis and spondylolisthesis and demonstrates typical location of spondylolysis. Greater laminectomy width has been associated with occurrence of acquired spondylolysis and spondylolisthesis.


Risk Prediction of Emergency Department Visits in Patients With Lung Cancer Using Machine Learning: Retrospective Observational Study.
Patients with lung cancer are among the most frequent visitors to emergency departments due to cancer-related problems, and the prognosis for those who seek emergency care is dismal. Given that patients with lung cancer frequently visit health care facilities for treatment or follow-up, the ability to predict emergency department visits based on clinical information gleaned from their routine visits would enhance hospital resource utilization and patient outcomes. This study proposed a machine learning-based prediction model to identify risk factors for emergency department visits by patients with lung cancer. This was a retrospective observational study of patients with lung cancer diagnosed at Seoul National University Bundang Hospital, a tertiary general hospital in South Korea, between January 2010 and December 2017. The primary outcome was an emergency department visit within 30 days of an outpatient visit. This study developed a machine learning-based prediction model using a common data model. In addition, the importance of features that influenced the decision-making of the model output was analyzed to identify significant clinical factors. The model with the best performance demonstrated an area under the receiver operating characteristic curve of 0.73 in its ability to predict the attendance of patients with lung cancer in emergency departments. The frequency of recent visits to the emergency department and several laboratory test results that are typically collected during cancer treatment follow-up visits were revealed as influencing factors for the model output. This study developed a machine learning-based risk prediction model using a common data model and identified influencing factors for emergency department visits by patients with lung cancer. The predictive model contributes to the efficiency of resource utilization and health care service quality by facilitating the identification and early intervention of high-risk patients. This study demonstrated the possibility of collaborative research among different institutions using the common data model for precision medicine in lung cancer.


Diversity of the KIR gene cluster in an urban Brazilian population.
The activity of natural killer cells depends on the balance between activating and inhibitory signals coming from their receptors. Among these are the killer cell immunoglobulin-like receptors (KIR) that recognize specific HLA class I allotypes. Here we characterized KIR genetic diversity and their HLA ligands in the population of Curitiba, Paraná State (n = 164), and compared it with other worldwide populations. The distribution of 2DL4 alleles was also analyzed. The Curitiba population did not differ significantly from European and Euro-descendant populations, but as an admixed population showed higher genetic diversity. We found 27 KIR profiles, many of them uncommon in European populations, in agreement with the elevated historically recent gene flow in the study population. The frequencies of KIR genes and their respective HLA ligands were distributed independently and none of the analyzed individuals lacked functional KIR-HLA ligand combinations. KIR gene frequencies of 33 worldwide populations were consistent with geographic and ethnic distribution, in agreement with demography being the major factor shaping the observed gene content diversity of the KIR locus.


Oestrogen increases haematopoietic stem-cell self-renewal in females and during pregnancy.
Sexually dimorphic mammalian tissues, including sexual organs and the brain, contain stem cells that are directly or indirectly regulated by sex hormones. An important question is whether stem cells also exhibit sex differences in physiological function and hormonal regulation in tissues that do not show sex-specific morphological differences. The terminal differentiation and function of some haematopoietic cells are regulated by sex hormones, but haematopoietic stem-cell function is thought to be similar in both sexes. Here we show that mouse haematopoietic stem cells exhibit sex differences in cell-cycle regulation by oestrogen. Haematopoietic stem cells in female mice divide significantly more frequently than in male mice. This difference depends on the ovaries but not the testes. Administration of oestradiol, a hormone produced mainly in the ovaries, increased haematopoietic stem-cell division in males and females. Oestrogen levels increased during pregnancy, increasing haematopoietic stem-cell division, haematopoietic stem-cell frequency, cellularity, and erythropoiesis in the spleen. Haematopoietic stem cells expressed high levels of oestrogen receptor-α (ERα). Conditional deletion of ERα from haematopoietic stem cells reduced haematopoietic stem-cell division in female, but not male, mice and attenuated the increases in haematopoietic stem-cell division, haematopoietic stem-cell frequency, and erythropoiesis during pregnancy. Oestrogen/ERα signalling promotes haematopoietic stem-cell self-renewal, expanding splenic haematopoietic stem cells and erythropoiesis during pregnancy.


Potential interference of leucite crystallization during porcelain thermal expansion measurement.
The elevated temperatures and slow heating and cooling rates associated with a thermal expansion run provide a thermal treatment comparable to those previously shown to produce leucite crystallization in dental PFM porcelains. The purpose of this study was to determine whether the exposure of dental porcelains to the time-temperature profile of a thermal expansion run could result in leucite crystallization, a potential interference with the accurate determination of the thermal expansion coefficient. A total of 112 thermal expansion specimens were prepared, 16 each of six commercially available body porcelains, as well as the "Component No. 1" (leucite-containing) frit of the Weinstein et al. (1962) patent. In a randomized design, the leucite contents of specimens subjected to a 3 degrees C/min thermal expansion run were compared to those of untreated controls. The leucite content of each specimen was determined via quantitative X-ray powder diffraction using an Al2O3 internal standard. One-tailed t-tests were applied to determine whether the dilatometer thermal treatment produced a statistically significant increase in leucite content. The heat treatment afforded by a 3 degrees C/min dilatometer run was not sufficient to induce measurable leucite crystallization in any of the porcelains examined in this study. The crystallization of leucite during a dilatometer heating run can be ruled out as a possible interference with the determination of the coefficient of thermal expansion of dental porcelain.


Alcohol consumption in young adults and incident hypertension: 20-year follow-up from the Coronary Artery Risk Development in Young Adults Study.
The relation between alcohol consumption and incident hypertension is unclear, and most observational studies have not accounted for socioeconomic factors. This study examined the association between alcohol consumption in a diverse group of young adults and incident hypertension over 20 years. Participants (n = 4,711) were from the Coronary Artery Risk Development in Young Adults Study cohort, recruited in 1985 (aged 18-30 years) from Birmingham, Alabama; Chicago, Illinois; Minneapolis, Minnesota; and Oakland, California. The 20-year incidence of hypertension for never, former, light, moderate, and at-risk drinkers was 25.1%, 31.8%, 20.9%, 22.2%, and 18.8%, respectively (P < 0.001). Race, gender, age, family history of hypertension, body mass index, income, education, and difficulty paying for basics and medical care were associated with hypertension. Adjustment using Cox proportional hazard models revealed no association between baseline alcohol consumption and incident hypertension, except among European-American women in whom any current alcohol consumption was associated with lower risk of incident hypertension. The lack of association between alcohol and hypertension in the majority of this socioeconomically diverse cohort is not definitive. Future studies should include social factors, such as income and education, and consider additional characteristics that may modify or confound associations between alcohol and blood pressure.


Referrals of patients by family physicians to consultants: a survey of the Israeli Family Practice Research Network.
We aimed to analyse factors influencing referral of patients by primary care physicians to specialist consultants at the beginning of the era of direct access to specialists in Israel. We carried out a study of referrals by family physicians to specialists over a continuous period of 3 months. Twenty-four certified family physicians filled in a questionnaire detailing referrals during the study period. All patients were referred for consultation to a specialist at regional speciality clinics or hospital out-patient departments. Ten physicians met the study conditions. In 1140 of 10896 (10.5%) visits, patients were referred to specialist consultants. The percentage of referral ranged from 7.4 to 15.9%. The difference between the physicians with the lowest and highest rates of referral was statistically significant (P < 0.0001). The variance in referral rates was not explained by significant differences in physician or practice variables. The types of specialists to whom the most referrals were made were orthopaedic surgeons, ophthalmologists, dermatologists, ear, nose and throat, general surgeons and plastic surgeons. There was a significant correlation between the type of specialist and the age of the patient. Older patients were referred more frequently to urologists, cardiologists and ophthalmologists, while younger patients were referred more frequently to ear, nose and throat specialists and gynaecologists (P < 0.01). The results of this study can be used as an aid for decision makers in the health services for determining policy. Direct access to some specialties might be appropriate, but not to all. Adoption of a policy based on these findings could lead to reduced health care costs by reducing the burden on hospital emergency rooms. It might also increase patient satisfaction in that the patients will have greater freedom of choice. On the other hand, more appropriate training of family physicians and more extensive self- and peer-quality assurance will increase the primary physician's knowledge and ability to diagnose and treat a broad range of problems and improve the level of care.


Assessing the impact of rising child poverty on the unprecedented rise in infant mortality in England, 2000-2017: time trend analysis.
To determine whether there were inequalities in the sustained rise in infant mortality in England in recent years and the contribution of rising child poverty to these trends. This is an analysis of trends in infant mortality in local authorities grouped into five categories (quintiles) based on their level of income deprivation. Fixed-effects regression models were used to quantify the association between regional changes in child poverty and regional changes in infant mortality. 324 English local authorities in 9 English government office regions. Live-born children under 1 year of age. Infant mortality rate, defined as the number of deaths in children under 1 year of age per 100 000 live births in the same year. The sustained and unprecedented rise in infant mortality in England from 2014 to 2017 was not experienced evenly across the population. In the most deprived local authorities, the previously declining trend in infant mortality reversed and mortality rose, leading to an additional 24 infant deaths per 100 000 live births per year (95% CI 6 to 42), relative to the previous trend. There was no significant change from the pre-existing trend in the most affluent local authorities. As a result, inequalities in infant mortality increased, with the gap between the most and the least deprived local authority areas widening by 52 deaths per 100 000 births (95% CI 36 to 68). Overall from 2014 to 2017, there were a total of 572 excess infant deaths (95% CI 200 to 944) compared with what would have been expected based on historical trends. We estimated that each 1% increase in child poverty was significantly associated with an extra 5.8 infant deaths per 100 000 live births (95% CI 2.4 to 9.2). The findings suggest that about a third of the increases in infant mortality between 2014 and 2017 can be attributed to rising child poverty (172 deaths, 95% CI 74 to 266). This study provides evidence that the unprecedented rise in infant mortality disproportionately affected the poorest areas of the country, leaving the more affluent areas unaffected. Our analysis also linked the recent increase in infant mortality in England with rising child poverty, suggesting that about a third of the increase in infant mortality from 2014 to 2017 may be attributed to rising child poverty.


Signaling mechanisms involved in protease-activated receptor-1-mediated interleukin-6 production by human gingival fibroblasts.
Human gingival fibroblasts (HGFs) express protease-activated receptor-1 (PAR-1) at high levels. In cultured HGFs, we studied the signaling pathway of thrombin-induced interleukin-6 (IL-6) production. The PAR-1 agonist peptide SFLLRN mimicked the thrombin-induced IL-6 production in the presence of amastatin, an aminopeptidase inhibitor. Thrombin or a combination of SFLLRN and amastatin also strikingly induced the expression of IL-6 mRNA. Although continuous exposure of HGFs to thrombin rapidly desensitized Ca(2+) signaling, the cells did not lose their ability to produce IL-6 in response to thrombin. Similarly, although treatment of HGFs with BAPTA-AM [1,2-bis(O-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester], an intracellular Ca(2+) chelator, markedly attenuated the thrombin-induced increase in intracellular Ca(2+) concentration, the same treatment did not suppress the thrombin-induced IL-6 production. However, thrombin-induced IL-6 production was strongly inhibited by the p38 mitogen-activated protein (MAP) kinase and tyrosine kinase inhibitors, and Western blotting analyses showed that thrombin stimulates p38 MAP kinase phosphorylation. Specific inhibitors that inhibit extracellular signal-regulated kinase 1/2 kinase, phosphatidylinositol 3-kinase, and RhoA kinase also partially suppressed the thrombin-induced IL-6 production, but the effects were smaller than those of the p38 MAP and tyrosine kinase inhibitors. Thus, thrombin induces HGFs to produce IL-6 by activating PAR-1, and the tyrosine kinase- and p38 MAP kinase-dependent pathways, rather than the Ca(2+) signaling pathway, may play a crucial role in the IL-6 production.


Inhibition of allergen-induced basophil activation by ASM-024, a nicotinic receptor ligand.
Nicotinic acetylcholine receptors (nAChRs) were identified on eosinophils and shown to regulate inflammatory responses, but nAChR expression on basophils has not been explored yet. We investigated surface receptor expression of nAChR α4, α7 and α1/α3/α5 subunits on basophils. Furthermore, we examined the effects of ASM-024, a synthetic nicotinic ligand, on in vitro anti-IgE and in vivo allergen-induced basophil activation. Basophils were enriched from the peripheral blood of allergic donors and the expression of nAChR subunits and muscarinic receptors was determined. Purified basophils were stimulated with anti-IgE in the presence of ASM-024 with or without muscarinic or nicotinic antagonists for the measurement of CD203c expression and histamine release. The effect of 9 days of treatment with 50 and 200 mg ASM-024 on basophil CD203c expression was examined in the blood of mild allergic asthmatics before and after allergen inhalation challenge. nAChR α4, α7 and α1/α3/α5 receptor subunit expression was detected on basophils. Stimulation of basophils with anti-IgE increased CD203c expression and histamine release, which was inhibited by ASM-024 (10(-5) to 10(-)(3) M, p < 0.05). The effect of ASM-024 was reversed in the presence of muscarinic and nicotinic antagonists. In subjects with mild asthma, ASM-024 inhalation significantly inhibited basophil CD203c expression measured 24 h after allergen challenge (p = 0.03). This study shows that ASM-024 inhibits IgE- and allergen-induced basophil activation through both nicotinic and muscarinic receptors, and suggests that ASM-024 may be an efficacious agent for modulating allergic asthma responses.


Epigenome Mapping Identifies Tumor-Specific Gene Expression in Primary Rectal Cancer.
Epigenetic alterations play a central role in cancer development and progression. The acetylation of histone 3 at lysine 27 (H3K27ac) specifically marks active genes. While chromatin immunoprecipitation (ChIP) followed by next-generation sequencing (ChIP-seq) analyses are commonly performed in cell lines, only limited data are available from primary tumors. We therefore examined whether cancer-specific alterations in H3K27ac occupancy can be identified in primary rectal cancer. Tissue samples from primary rectal cancer and matched mucosa were obtained. ChIP-seq for H3K27ac was performed and differentially occupied regions were identified. The expression of selected genes displaying differential occupancy between tumor and mucosa were examined in gene expression data from an independent patient cohort. Differential expression of four proteins was further examined by immunohistochemistry. ChIP-seq for H3K27ac in primary rectal cancer and matched mucosa was successfully performed and revealed differential binding on 44 regions. This led to the identification of genes with increased H3K27ac, i.e., RIPK2, FOXQ1, KRT23, and EPHX4, which were also highly upregulated in primary rectal cancer in an independent dataset. The increased expression of these four proteins was confirmed by immunohistochemistry. This study demonstrates the feasibility of ChIP-seq-based epigenome mapping of primary rectal cancer and confirms the value of H3K27ac occupancy to predict gene expression differences.


Chest pain units: A necessity or only a name to encourage patients.
Acute chest pain is a common symptom among patients presenting to emergency wards. Identification and admission of patients with real acute coronary syndrome and preventing the hospitalization of people with false diagnosis of coronary syndrome are the most important tasks in emergency wards. The purpose of this study was to investigate the usefulness of designing a special chest pain unit in emergency department of Imam Khomeini Hospital (Ahvaz, Iran). The patients with markers of ongoing cardiac ischemia underwent selective coronary angiography. The chest pain unit protocol was applied to selected patients with no definite evidence of acute coronary syndrome or alternative pathology. The protocol consisted of twelve hours of observation and serial 12-lead electrocardiography, transthoracic echocardiography, and biochemical testing followed by an exercise treadmill test. We compared the number of patients who were discharged after work up, discharged themselves against medical advice, admitted at coronary care unit (CCU), underwent invasive procedures or died between 2007 and 2010. During 2010, 43% of patients were discharged after evaluation in the chest pain unit. In 2007 however, 26% were discharged following traditional assessments. The admission rate increased from 23% in 2007 to 36% in 2010. The percentage of patients who discharged themselves against medical advice decreased from 37% in 2007 to 14% in 2010. There was not a statistically significant difference between mortality rates in 2007 and 2010. Providing a special chest pain unit in emergency ward in our condition is helpful. It reduces unnecessary admissions and improves patient satisfaction.


Tissue-Welding Device: Considerable Advantages for Spleen Surgery Based on Histological and Cardiorespiratory Investigation.
During splenic surgery, it is important to control blood loss and the potential risk of cardiac arrhythmia. The best way to prevent complications from surgery is to use the appropriate surgical devices; however, there is no guideline for the use of specific surgical devices for spleen incision. Therefore, the aim of this research was to compare the interactions of various surgical devices with spleen tissue, their cardiorespiratory effects during incision, and subsequent spleen surgical wound healing. A total of 75 rabbits were included in the study. CO2 laser (n = 15), radiofrequency device (n = 15), electrocoagulator (n = 15), tissue-welding device (n = 15), and scalpel (n = 15) were used to make incisions in rabbits' spleens. Spleen biopsies of the incision area were taken from each animal at the day 0, 7, and 14 after surgery. Contactless thermography was performed during surgery. Suturing was not used after incision with the tissue-welding device, but incisions made by other surgical devices were sutured. The results showed that the width of spleen necrosis differed significantly between the various surgical devices used on spleen tissues. There was a positive, strong, and linear association between necrosis width and the tissue temperature of cutting edges. Significant increases in the heart rate were observed during spleen surgery performed with laser, scalpel, and radiofrequency devices. In conclusion, the tissue-welding device confers a significant advantage in spleen surgery, as there is neither a need for sutures nor a significant deviation in the heart rate.


Cancer in people with AIDS in New Jersey.
A total of 2,460 persons with AIDS (PWA) and cancer were identified--2,159 men and 301 women--among 22,382 men and 7,931 women reported with AIDS from 1979-1995 in New Jersey. Of these, 50% were diagnosed with Kaposi's sarcoma (KS), 33% with non-Hodgkin's lymphoma (NHL), and the remainder with other cancers, including lung and bronchus, oral, cervical, Hodgkin's disease, and digestive other than anal. The proportions of PWA who also had cancer, specifically KS and NHL, varied significantly by gender, race, and mode of HIV transmission. These findings are discussed in terms of risk factors in addition to immunosuppression that may account for the higher rates of cancer among PWA than the general population. Cancer prevention and detection measures likely will become more important as PWA survive longer.


Aberrant proliferation in CXCR7+ endothelial cells via degradation of the retinoblastoma protein.
Angiogenesis is a critical factor in the growth and dissemination of solid tumors. Indeed, tumor vasculature is abnormal and contributes to the development and spread of malignancies by creating a hostile microenvironment. The alternative SDF-1/CXCL12 receptor, CXCR7, is frequently and specifically expressed in tumor-associated vessels. In this study, we examine the role of endothelium-expressed CXCR7 in tumor vascular dysfunction by specifically examining the contribution of CXCR7 to endothelial cell (EC) proliferation. We demonstrate that CXCR7 expression is sufficient to drive post-confluent growth in EC cultures. Further, we provide a novel mechanism for CXCR7-mediated proliferation via proteasomal degradation of the tumor suppressor protein Rb. These findings identify a heretofore unappreciated role for CXCR7 in vascular dysfunction and confirm this receptor as a plausible target for anti-tumor therapy.


Cryotherapy causes extensive breakdown of the blood-retinal barrier. A comparison with argon laser photocoagulation.
Using computerized vitreous fluorophotometry (VFP) in pigmented rabbits, we examined two modalities frequently used in retinal reattachment surgery, cryotherapy and argon laser photocoagulation, for their effect on the blood-retinal barrier. The VFP readings were taken 2 mm posterior to the lens one hour after intravenous injection of 14 mg/kg of fluorescein sodium. After baseline fluorophotometry readings, rabbits were treated with confluent cryotherapy over the inferior 180 degrees in one eye and with confluent laser over an equivalent area of retina in the other eye. The VFP readings were taken 2, 4, 6, and 15 days after treatment. By day 6, the VFP reading had risen from a pretreatment value of 6.4 +/- 0.4 ng/mL to 41.8 +/- 7.9 ng/mL in the cryotherapy-treated eyes as opposed to 15.5 +/- 3.1 ng/mL in the laser-treated eyes. By day 15, the readings were almost back to baseline, and there was no significant difference between the two treatment groups. These data suggest that there is a significant breakdown of the blood-retinal barrier with both modalities, but that it is considerably more severe with cryotherapy.


Neuroprotective role of Ginkgo biloba against cognitive deficits associated with Bisphenol A exposure: An animal model study.
Our study aimed to elucidate to what extent Ginkgo biloba (Gb) can protect rats from cognitive deficits induced by exposure to Bisphenol A (BPA) at high dose. Therefore, sixty male Wistar rats were randomly divided into four groups of 15 animals in each group: Vehicle group, Gb-control group, BPA-exposed group and Gb pre-treated group. All administrations were given daily by an oral gavage once a day for eight weeks. Cognitive function was assessed using Morris water maze; Y-maze and Novel object recognition tasks. Additionally, hippocampal levels of DA, NE and 5-HT were measured. BPA-induced oxidative stress was evaluated by determining SOD activity, NO and MDA levels in rat hippocampus as well as level of circulating adiponectin. Moreover, histopathological changes in CA3 region of rat hippocampus and immunohistochemical expression of NF-κB and Caspase-3 were investigated. We found that Gb pretreatment significantly improved cognitive performance; may be via increasing hippocampal levels of estrogen-dependent biogenic amines. At the same time, Gb could strictly control BPA-induced oxidative stress by improving SOD activity and adiponectin level with decrease in NO and MDA levels. Lastly, Gb alleviated the histopathological injuries induced by BPA and inhibited NF-κB and caspase-3 activation. In conclusion, our results suggested that Gb has potential to ameliorate BPA-induced hippocampal neuronal damage and subsequent cognitive deficits through mechanisms involving its ability to enhance the release of biogenic amines as well as its antioxidant and adiponectin pro-secretory effects.


Structure of the primed state of the ATPase domain of chromatin remodeling factor ISWI bound to the nucleosome.
ATP-dependent chromatin remodeling factors of SWI/SNF2 family including ISWI, SNF2, CHD1 and INO80 subfamilies share a conserved but functionally non-interchangeable ATPase domain. Here we report cryo-electron microscopy (cryo-EM) structures of the nucleosome bound to an ISWI fragment with deletion of the AutoN and HSS regions in nucleotide-free conditions and the free nucleosome at ∼ 4 Å resolution. In the bound conformation, the ATPase domain interacts with the super helical location 2 (SHL 2) of the nucleosomal DNA, with the N-terminal tail of H4 and with the α1 helix of H3. Density for other regions of ISWI is not observed, presumably due to disorder. Comparison with the structure of the free nucleosome reveals that although the histone core remains largely unchanged, remodeler binding causes perturbations in the nucleosomal DNA resulting in a bulge near the SHL2 site. Overall, the structure of the nucleotide-free ISWI-nucleosome complex is similar to the corresponding regions of the recently reported ADP bound ISWI-nucleosome structures, which are significantly different from that observed for the ADP-BeFx bound structure. Our findings are relevant to the initial step of ISWI binding to the nucleosome and provide additional insights into the nucleosome remodeling process driven by ISWI.


Theoretical structure-activity studies of beta-carboline analogs. Requirements for benzodiazepine receptor affinity and antagonist activity.
The techniques of theoretical chemistry have been used to elucidate the molecular properties and modes of receptor binding that modulate receptor affinity and antagonist activity of the beta-carbolines, a class of potent benzodiazepine antagonists. Six analogs were chosen in order to investigate the role of the amine (NH) group, the aromatic nitrogen, and the C3-substituent in determining receptor affinities. Electrostatic potential mapping and characterization of explicit drug-receptor interactions have led to the hypothesis that simultaneous interaction of a model cationic arginine site with the N2 and C3-substituents could play a key role in determining receptor affinities. The electron-withdrawing effects of C3-substituents on the amine nitrogen appear less important, though interactions of these groups with an anionic glutamate or aspartate site could also occur at the receptor. Similarly, stacking interactions with neutral or cationic aromatic residues such as tryptophan or protonated histidine could occur, but do not appear to be determinants of the relative receptor affinity of the beta-carbolines.


Aberrant Patterns of Key Epithelial Basement Membrane Components in Keratoconus.
In the cornea, the epithelial basement membrane (BM) plays an important role in maintaining corneal integrity and homeostasis. Aberrations in this vital structure are associated with several corneal pathologies including keratoconus. The aim of this study was to investigate the expression of key structural components of the epithelial BM in keratoconic corneas and to identify and describe any aberrant patterns. Immunohistochemical labeling of key BM components including fibronectin, laminin, and type IV and VII collagen was performed in healthy and keratoconic corneas. Clear changes in the BM components in the keratoconic corneas were seen with the key structural components either being absent or forming a discontinuous pattern. Another aberrant pattern, the expression of BM proteins, particularly fibronectin, laminin, and type IV collagen, in the anterior stroma of keratoconic corneas was also observed. These results indicate the activation of keratocytes into the fibroblast and myofibroblast wound phenotypes and the potential source of corneal scarring commonly observed in keratoconic corneas. Our data also support the hypothesis of dysregulated collagen synthesis and breakdown in the keratoconic cornea, in particular, the BM, and suggest a role for the BM in initiation and progression of keratoconus.


Recovery of amplitude integrated electroencephalographic background patterns within 24 hours of perinatal asphyxia.
To assess the time course of recovery of severely abnormal initial amplitude integrated electroencephalographic (aEEG) patterns (flat trace (FT), continuous low voltage (CLV), or burst suppression (BS)) in full term asphyxiated neonates, in relation to other neurophysiological and neuroimaging findings and neurodevelopmental outcome. A total of 190 aEEGs of full term infants were reviewed. The neonates were admitted within 6 hours of birth to the neonatal intensive care unit because of perinatal asphyxia, and aEEG recording was started immediately. In all, 160 infants were included; 65 of these had an initial FT or CLV pattern and 25 an initial BS pattern. Neurodevelopmental outcome was assessed using a full neurological examination and the Griffiths' mental developmental scale. In the FT/CLV group, the background pattern recovered to continuous normal voltage within 24 hours in six of the 65 infants (9%). All six infants survived the neonatal period; one had a severe disability, and five were normal at follow up. In the BS group, the background pattern improved to normal voltage in 12 of the 25 infants (48%) within 24 hours. Of these infants, one died, five survived with moderate to severe disability, two with mild disability, and four were normal. The patients who did not recover within 24 hours either died in the neonatal period or survived with a severe disability. In this study there was a small group of infants who presented with a severely abnormal aEEG background pattern within six hours of birth, but who achieved recovery to a continuous normal background pattern within the first 24 hours. Sixty one percent of these infants survived without, or with a mild, disability.


A chaperone function of NO CATALASE ACTIVITY1 is required to maintain catalase activity and for multiple stress responses in Arabidopsis.
Catalases are key regulators of reactive oxygen species homeostasis in plant cells. However, the regulation of catalase activity is not well understood. In this study, we isolated an Arabidopsis thaliana mutant, no catalase activity1-3 (nca1-3) that is hypersensitive to many abiotic stress treatments. The mutated gene was identified by map-based cloning as NCA1, which encodes a protein containing an N-terminal RING-finger domain and a C-terminal tetratricopeptide repeat-like helical domain. NCA1 interacts with and increases catalase activity maximally in a 240-kD complex in planta. In vitro, NCA1 interacts with CATALASE2 (CAT2) in a 1:1 molar ratio, and the NCA1 C terminus is essential for this interaction. CAT2 activity increased 10-fold in the presence of NCA1, and zinc ion binding of the NCA1 N terminus is required for this increase. NCA1 has chaperone protein activity that may maintain the folding of catalase in a functional state. NCA1 is a cytosol-located protein. Expression of NCA1 in the mitochondrion of the nca1-3 mutant does not rescue the abiotic stress phenotypes of the mutant, while expression in the cytosol or peroxisome does. Our results suggest that NCA1 is essential for catalase activity.


Correlates of traumatic brain injury among juvenile offenders: a multi-site study.
There is some evidence that antisocial individuals, including young delinquents, are significantly more likely than people in the general population to incur a traumatic brain injury (TBI). Extant studies are hampered by methodological limitations, such as small sample sizes, lack of control for confounding effects, and use of single sites and may lack representativeness. The hypothesis for this study is that young offenders with a history of TBI will not only be at higher risk of impulsivity and negative emotionality than their non-injured peers but also that those with TBI will have had more previous victimisation experiences. Data from two sites (Philadelphia, PA and Phoenix, AZ) in a substantial longitudinal, prospective study - the Pathways to Desistance Study - were used to compare young people (average age 16 years) who reported TBI with those who did not. Independent variables were behavioural, criminogenic and psychosocial measures. Male youths were about twice as likely as young females to report TBI. Such injury was associated with higher impulsivity and negative emotion ratings, even after allowing for potentially confounding factors, including sex. In addition, TBI was independently associated with self-reported experience of victimisation. This extends the generalisability of earlier suggestions of a relationship between TBI and offending, and various factors thought likely to mediate the relationship. The implications are, thus, that it is likely to be of practical value to screen young people who get into the criminal justice system for a history of TBI, and thus to allow for more specific tailoring of interventions to reduce the range of associated problems, including likely reoffending.


In patients with LTP syndrome food-specific IgE show a predictable hierarchical order.
Lipid transfer protein (LTP) is a widely cross-reacting allergen in plant foods. To assess whether IgE to vegetable foods show predictable trends in LTP allergic patients. Clinical allergy to plant foods other than peach was sought in 15 consecutive peach-allergic adults monosensitized to LTP. IgE specific for peach, apple, hazelnut, walnut, peanut, lentil, maize, soybean, tomato, sesame, mustard melon, kiwi, and celery as well as to mugwort pollen was measured. Peach-specific IgE levels exceeded IgE to all other study foods. The higher were peach-specific IgE levels, the higher was the probability that other plant-derived foods scored positive. Mean IgE levels specific for all study foods were strongly correlated to peach specific IgE. Food-specific IgE followed a rather precise hierarchy, both in terms of number of positive in-vitro tests and of IgE levels, with apple at the second place after peach, followed by walnut, hazelnut, peanut, lentil, maize, soybean, tomato, kiwi, sesame, mustard, melon, and celery. Such hierarchy was not necessarily paralleled by clinical allergy as lentil, maize, and soybean scored positive in the majority of patients, but induced allergy in 0, 1, and 0 patients, respectively. IgE levels were not necessarily correlated with the severity of clinical allergy. Little or no IgE reactivity to mugwort pollen was found. In LTP syndrome, IgE reactivity to foods other than peach is in most cases predictable and follows a regular sequence that probably depends on the degree of homology with Pru p3. The reasons why some foods are tolerated by most patients despite elevated IgE reactivity remains to be elucidated.


Efficient parallelization of short-range molecular dynamics simulations on many-core systems.
This article introduces a highly parallel algorithm for molecular dynamics simulations with short-range forces on single node multi- and many-core systems. The algorithm is designed to achieve high parallel speedups for strongly inhomogeneous systems like nanodevices or nanostructured materials. In the proposed scheme the calculation of the forces and the generation of neighbor lists are divided into small tasks. The tasks are then executed by a thread pool according to a dependent task schedule. This schedule is constructed in such a way that a particle is never accessed by two threads at the same time. Benchmark simulations on a typical 12-core machine show that the described algorithm achieves excellent parallel efficiencies above 80% for different kinds of systems and all numbers of cores. For inhomogeneous systems the speedups are strongly superior to those obtained with spatial decomposition. Further benchmarks were performed on an Intel Xeon Phi coprocessor. These simulations demonstrate that the algorithm scales well to large numbers of cores.


Diffusion MRI quantifies early axonal loss in the presence of nerve swelling.
Magnetic resonance imaging markers have been widely used to detect and quantify white matter pathologies in multiple sclerosis. We have recently developed a diffusion basis spectrum imaging (DBSI) to distinguish and quantify co-existing axonal injury, demyelination, and inflammation in multiple sclerosis patients and animal models. It could serve as a longitudinal marker for axonal loss, a primary cause of permanent neurological impairments and disease progression. Eight 10-week-old female C57BL/6 mice underwent optic nerve DBSI, followed by a week-long recuperation prior to active immunization for experimental autoimmune encephalomyelitis (EAE). Visual acuity of all mice was assessed daily. Longitudinal DBSI was performed in mouse optic nerves at baseline (naïve, before immunization), before, during, and after the onset of optic neuritis. Tissues were perfusion fixed after final in vivo scans. The correlation between DBSI detected pathologies and corresponding immunohistochemistry markers was quantitatively assessed. In this cohort of EAE mice, monocular vision impairment occurred in all animals. In vivo DBSI detected, differentiated, and quantified optic nerve inflammation, demyelination, and axonal injury/loss, correlating nerve pathologies with visual acuity at different time points of acute optic neuritis. DBSI quantified, in the presence of optic nerve swelling, ~15% axonal loss at the onset of optic neuritis in EAE mice. Our findings support the notion that axonal loss could occur early in EAE mice. DBSI detected pathologies in the posterior visual pathway unreachable by optical coherence tomography and without confounding inflammation induced optic nerve swelling. DBSI could thus decipher the interrelationship among various pathological components and the role each plays in disease progression. Quantification of the rate of axonal loss could potentially serve as the biomarker to predict treatment outcome and to determine when progressive disease starts.


Variants in the degron of AFF3 are associated with intellectual disability, mesomelic dysplasia, horseshoe kidney, and epileptic encephalopathy.
The ALF transcription factor paralogs, AFF1, AFF2, AFF3, and AFF4, are components of the transcriptional super elongation complex that regulates expression of genes involved in neurogenesis and development. We describe an autosomal dominant disorder associated with de novo missense variants in the degron of AFF3, a nine amino acid sequence important for its binding to ubiquitin ligase, or with de novo deletions of this region. The sixteen affected individuals we identified, along with two previously reported individuals, present with a recognizable pattern of anomalies, which we named KINSSHIP syndrome (KI for horseshoe kidney, NS for Nievergelt/Savarirayan type of mesomelic dysplasia, S for seizures, H for hypertrichosis, I for intellectual disability, and P for pulmonary involvement), partially overlapping the AFF4-associated CHOPS syndrome. Whereas homozygous Aff3 knockout mice display skeletal anomalies, kidney defects, brain malformations, and neurological anomalies, knockin animals modeling one of the microdeletions and the most common of the missense variants identified in affected individuals presented with lower mesomelic limb deformities like KINSSHIP-affected individuals and early lethality, respectively. Overexpression of AFF3 in zebrafish resulted in body axis anomalies, providing some support for the pathological effect of increased amount of AFF3. The only partial phenotypic overlap of AFF3- and AFF4-associated syndromes and the previously published transcriptome analyses of ALF transcription factors suggest that these factors are not redundant and each contributes uniquely to proper development.


Use of a dual mobility socket to manage total hip arthroplasty instability.
Unconstrained tripolar hip implants provide an additional bearing using a mobile polyethylene component between the prosthetic head and the outer metal shell. Such a design increases the effective head diameter and therefore is an attractive option in challenging situations of unstable total hip arthroplasties. We report our experience with 54 patients treated using this dual mobility implant in such situations. We ascertained its ability to restore and maintain stability, and examined component loosening and component failure. At a minimum followup of 2.2 years (mean, 4 years; range, 2.2-6.8 years), one hip had redislocated 2 months postoperatively and was managed successfully without reoperation by closed reduction with no additional dislocation. Two patients required revision of the implant because of dislocation at the inner bearing. Technical errors were responsible for these failures. Three patients had reoperations for deep infections. The postoperative radiographs at latest followup showed very satisfactory osseointegration of the acetabular component because no radiolucent line or osteolysis was reported. Use of this unconstrained tripolar design was successful in restoring and maintaining hip stability. We observed encouraging results at short-term followup regarding potential for loosening or mechanical failures. Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence.


Leucine elicits myotube hypertrophy and enhances maximal contractile force in tissue engineered skeletal muscle in vitro.
The amino acid leucine is thought to be important for skeletal muscle growth by virtue of its ability to acutely activate mTORC1 and enhance muscle protein synthesis, yet little data exist regarding its impact on skeletal muscle size and its ability to produce force. We utilized a tissue engineering approach in order to test whether supplementing culture medium with leucine could enhance mTORC1 signaling, myotube growth, and muscle function. Phosphorylation of the mTORC1 target proteins 4EBP-1 and rpS6 and myotube hypertrophy appeared to occur in a dose dependent manner, with 5 and 20 mM of leucine inducing similar effects, which were greater than those seen with 1 mM. Maximal contractile force was also elevated with leucine supplementation; however, although this did not appear to be enhanced with increasing leucine doses, this effect was completely ablated by co-incubation with the mTOR inhibitor rapamycin, showing that the augmented force production in the presence of leucine was mTOR sensitive. Finally, by using electrical stimulation to induce chronic (24 hr) contraction of engineered skeletal muscle constructs, we were able to show that the effects of leucine and muscle contraction are additive, since the two stimuli had cumulative effects on maximal contractile force production. These results extend our current knowledge of the efficacy of leucine as an anabolic nutritional aid showing for the first time that leucine supplementation may augment skeletal muscle functional capacity, and furthermore validates the use of engineered skeletal muscle for highly-controlled investigations into nutritional regulation of muscle physiology.


Influence of Melatonin on Behavioral and Neurological Function of Rats with Focal Cerebral Ischemia-Reperfusion Injury via the JNK/FoxO3a/Bim Pathway.
To investigate the influence of melatonin on behavioral and neurological function of rats with focal cerebral ischemia-reperfusion injury via the JNK/FoxO3a/Bim pathway. One hundred and twenty healthy male SD rats were randomized into the model group (Model: the middle cerebral artery occlusion (MCAO) model was constructed and received an equal volume of normal saline containing 5% DMSO), sham operation group (Sham: received no treatment except normal feeding), and low, medium, and high dose of melatonin group (L-MT, M-MT, and H-MT intraperitoneally injected 10, 20, and 40 mg/kg melatonin 30 min after IR, respectively), with 24 rats in each group. Following 24 h of reperfusion, the rats in each of the above groups were tested for neurological deficit symptoms and behavioral changes to screen the rats included in the study. HE and TUNEL stainings were performed to observe pathological changes. Levels of oxidative stress-related indexes, inflammatory factor-related indexes, nuclear factor-κB p65 (NF-κB p65), and interferon-γ (IFN-γ) in the rat brain were measured by ELISA. The JNK/FoxO3a/Bim pathway-related proteins as well as Bcl-2, Caspase-3, and Bax were examined using Western blot. Detection of behavioral indicators showed that the MACO model was successfully constructed in rats. L-MT, M-MT, and L-MT groups presented reduced malondialdehyde (MDA), reactive oxygen species (ROS), tumor necrosis factor- (TNF-) α, interleukin- (IL-) 6, IL-1β, IFN-γ, NF-κB p65, and apoptosis compared with the Model group (P < 0.05), and the improvement degree was better in the M-MT group versus the L-HT group. Bcl-2 protein expression in the brain tissue of L-MT, M-MT, and H-MT groups increased significantly, while Bax, Caspase-3, p-JNK, p-FoxO3a, and Bim protein expression declined markedly, versus the Model group (P < 0.05). The changes of indexes were greater in the M-MT group compared with that in the L-MT group. No significant difference was observed in all the above indexes between the M-MT group and the H-MT group (P > 0.05). In the MACO rat model, melatonin can effectively reduce Bax and Caspase-3 levels by modulating the JNK/FoxO3a/Bim pathway, inhibit neuronal apoptosis, and alleviate neurological deficits by reducing the release of proinflammatory mediators, with anti-inflammatory and antioxidant effects. In addition, 20 mg/kg is the optimal melatonin concentration.