Facile, non-destructive characterization of 2d photonic crystals using UV-vis-spectroscopy.
We present a simple and non-destructive method for characterizing and quantifying the quality of two-dimensional (2D) close-packed arrays of submicron dielectric spheres. Utilizing radiative losses of photonic modes created by the 2D crystals into dielectric substrates we are able to monitor the quality of the particle monolayer during assembly and the size evolution of the individual particles during dry etching. Using an advanced interfacial assembly technique we prepare particle monolayers on glass and characterize the spectral behaviour of the radiative loss regarding different lattice constants, dielectric substrates and layer qualities. The effect of diameter reduction during dry etching is analysed and a simple model is proposed, which enables non-destructive, on spot characterization of the particle layer with sub-20 nm resolution using UV-vis spectroscopy.


A case series of giant molluscum contagiosum in an immigrant African pediatric population.
Giant molluscum contagiosum (MC) has a well-known association with human immunodeficiency virus and other immune deficiency states. Although rare, it can be seen in healthy immunocompetent children. We describe eight cases of giant MC in healthy, immunocompetent African immigrant children in the Columbus, Ohio area. This report describes the clinical characteristics, treatment, and course of giant MC in this patient population.


Self-medication for migraine: A nationwide cross-sectional study in Italy.
Headache disorders are considered the second leading cause of years lived with disability worldwide, and 90% of people have a headache episode at least once a year, thus representing a relevant public health priority. As the pharmacist is often the first and only point of reference for people complaining of headache, we carried out a survey in a nationwide sample of Italian pharmacies, in order to describe the distribution of migraine or non-migraine type headaches and medicines overuse among people entering pharmacies seeking for self-medication; and to evaluate the association, in particular of migraine, with socio-demographic and clinical characteristics, and with the pathway of care followed by the patients. A 14-item questionnaire, including socio-demographic and clinical factors, was administered by trained pharmacists to subjects who entered a pharmacy requesting self-medication for a headache attack. The ID Migraine™ Screener was used to classify headache sufferers in four classes. From June 2016 to January 2017, 4424 people have been interviewed. The prevalence of definite migraines was 40%, significantly higher among women and less educated people. About half of all headache sufferers and a third of migraineurs do not consider their condition as a disease and are not cared by any doctor. Among people seeking self-medication in pharmacies for acute headache attacks, the rate of definite or probable migraine is high, and a large percentage of them is not correctly diagnosed and treated. The pharmacy can be a valuable observatory for the study of headaches, and the first important step to improve the quality of care delivered to these patients.


The β3-integrin binding protein β3-endonexin is a novel negative regulator of hypoxia-inducible factor-1.
Integrins are multifunctional heterodimeric adhesion receptors that mediate the attachment between a cell and the extracellular matrix or other surrounding cells. In endothelial cells, integrins can modulate cell migration and motility. In particular, β3-integrin is expressed in angiogenic vessels. Signal transduction by β3-integrins requires the recruitment of intracellular signaling molecules. β3-endonexin is a highly spliced molecule that has been identified as a β3-integrin binding protein. β3-endonexin isoforms are expressed in endothelial cells and have been suggested to act as shuttle proteins between the membrane and the nucleus. However, their functional role in angiogenesis is unclear. In this study, we investigated whether β3-endonexin isoforms are involved in endothelial angiogenic processes under hypoxia. The overexpression of β3-endonexin isoforms decreased endothelial proliferation and tube formation under hypoxia, while the depletion of β3-endonexin by RNAi promoted angiogenic responses in vitro and in vivo. In hypoxia, β3-endonexin accumulated in the nucleus, and prevention of this response by depletion of β3-endonexin increased hypoxic activation and induction of the hypoxia-inducible factor (HIF)-1 and its target genes VEGF and PAI-1. β3-endonexin diminished nuclear factor kappa B (NFκB) activation and decreased NFκB binding to the HIF-1α promoter under hypoxia, subsequently diminishing NFκB-dependent transcription of HIF-1α under hypoxia. Our results indicate for the first time that the overexpression of β3-endonexin can decrease hypoxic induction and activation of HIF-1α and can prevent hypoxic endothelial proliferation and angiogenic responses. β3-endonexin can act as a novel anti-angiogenic factor specifically in the response to hypoxia due to its negative impact on the activation of HIF-1.


Oral Azelaic Acid Ester Decreases Markers of Insulin Resistance in Overweight Human Male Subjects.
Insulin resistance (IR) is linked to increased risk of cardiovascular disease and cancer. We examined safety and efficacy of the natural product diethyl azelate (DEA) in overweight males with a varying degree of IR. Seventeen subjects [age 18-42, hemoglobin A1c (A1c) of 5.2-6.2%] received orally 1 mg/kg DEA daily for 21 days. Blood plasma glucose, insulin and lipid levels were assessed before and after treatment. DEA was well tolerated without hypoglycemia or adverse effects except transient diarrhea (n=1). DEA significantly reduced fasting glucose by 6.06 mg/dl (n=8) and insulin by 37.8% (n=8) in subjects with IR and/or A1c ≥5.6%. Furthermore, it improved cholesterol/HDL, LDL/HDL, and non-cholesterol HDL/HDL by 5.4, 6.5, and 6.6%, respectively in all subjects, and by 8.0, 9.8, and 9.8%, respectively in 9 subjects with A1c ≥5.6%. DEA efficacy correlates with the degree of IR. DEA holds promise as a novel treatment for the management of IR.


Guideline on the diagnosis and treatment of primary idiopathic clubfoot.
- A delegation of 6 pediatric orthopedic surgeons from the Dutch Orthopedic Association (NOV) and 2 members of the board of the Dutch Parents' Association for children with clubfoot created the guideline "The diagnosis and treatment of primary idiopathic clubfeet" between April 2011 and February 2014. The development of the guideline was supported by a professional methodologist from the Dutch Knowledge Institute of Medical Specialists. This evidence-based guideline process was new and unique, in the sense that the process was initiated by a parents' association. This is the first official guideline in pediatric orthopedics in the Netherlands, and to our knowledge it is also the first evidence-based guideline on clubfoot worldwide. The guideline was developed in accordance with the criteria of the international AGREE instrument (AGREE II: Appraisal of Guidelines for Research and Evaluation II). The scientific literature was searched and systematically analyzed. In the second phase, conclusions and recommendations in the literature were formulated according to the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) method. Recommendations were developed considering the balance of benefits and harms, the type and quality of evidence, the values and preferences of the people involved, and the costs. The guideline is a solid foundation for standardization of clubfoot treatment in the Netherlands, with a clear recommendation of the Ponseti method as the optimal method of primary clubfoot treatment. We believe that the format used in the current guideline sets a unique example for guideline development in pediatric orthopedics that may be used worldwide. Our format ensured optimal collaboration between medical specialists and parents, and resulted in an important change in clubfoot care in the Netherlands, to the benefit of medical professionals as well as parents and patients. In this way, it is possible to improve professional collaboration between medical specialists and parents, resulting in an important change in clubfoot care in the Netherlands that will benefit medical professionals, parents, and patients. The guideline was published online, and is freely available from the Dutch Guideline Database ( www.richtlijnendatabase.nl ).


Alcohol and GABA-benzodiazepine receptor function.
gamma-Aminobutyric acid (GABA)A is a major inhibitory neurotransmitter in the mammalian CNS. GABAA ergic synapse is also an important site of action for a variety of centrally acting drugs, including benzodiazepines and barbiturates. Several lines of electrophysiological, behavioral, and biochemical studies implicate GABAA ergic synapse in the actions of alcohol. In electrophysiological studies, alcohol has been reported to enhance GABA-mediated responses in cortical neurons, spinal cord and substantia nigra, albeit, negative results have also been reported. In behavioral studies, GABAmimetics enhance alcohol's effects on motor coordination, while GABA antagonists have the opposite effect. In drug-combination studies, subeffective doses of alcohol, in combination with subeffective doses of other GABAmimetics, potentiate each other's effect in several seizure models. In functional studies, alcohol has been reported to potentiate GABA receptor-mediated 36Cl-flux in microsacs, neurosynaptosomes, and cultured spinal cord neurons at pharmacologically relevant concentrations. The potentiating effect of alcohol is blocked by GABA antagonists and the inverse agonists of the benzodiazepine receptor site. Taken together, these studies indicate that some of the central effects of alcohol are mediated via facilitation of GABAAergic transmission.


Double-negative (CD4-CD8-) TCRalphabeta+ cells in patients with systemic lupus erythematosus.
Massive accumulation of CD4-CD8-TCRalphabeta+ cells in secondary lymphoid organs is characteristic of lupus-prone MRL/lpr mice. However, the role of these double negative T cells (DNT) in human lupus patients receive only limited attention. Herein, we investigate the frequency of DNT in the peripheral blood mononuclear cells of forty seven Chinese patients with systemic lupus erythematosus (SLE) and forty four normal individuals. DNT were measured with dual-fluorescence flow cytometry. The results showed that DNT only constituted a very minor subset of lymphocytes both in patients and normals, it normally did not exceed 2% of the lymphocyte population. Compared with normal subjects, patients with SLE had slightly increased levels of DNT within the total lymphocyte population (0.66+/-0.45% vs 0.51+/-0.33%) or within TCRalphabeta+ population (1.14+/-0.88% vs 0.88+/-0.54%). The difference, however, did not reach statistical significance. The levels of DNT correlated neither with the titers of anti-DNA antibodies in sera nor with the presence of active and severe lupus nephritis in SLE patients. Longitudinal follow-up of six patients at the stages of active and inactive nephritis revealed similar levels of DNT in the same individual. The preliminary results suggest that circulating DNT do not appear to play a critical role in Chinese patients with SLE.


Identification of Tumor Antigens and Design of mRNA Vaccine for Colorectal Cancer Based on the Immune Subtype.
mRNA vaccines have become a promising alternative to conventional cancer immunotherapy approaches. However, its application on colorectal cancer (CRC) remains poorly understood. We herein identified potential antigens for designing an effective mRNA vaccine, further to build an immune landscape for the accurate selection of patients for mRNA vaccine therapy. Raw transcriptome data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases were retrieved. Consensus clustering algorithm was applied to divide the CRC samples into four immune subtypes. Immunogenomics analysis was further integrated to characterize the immune microenvironment of each immune subtype. Six tumor antigens were found to be associated with poor prognosis and infiltration of antigen-presenting cells (APCs) in CRC patients. Furthermore, each of the immune subtypes showed differential cellular and molecular features. The IS2 and IS4 exhibited significantly improved survival and higher immune cell infiltration compared with IS1 and IS3. Immune checkpoint molecules and human leukocyte antigen also showed significant differential expression in four immune subtypes. Moreover, we performed graph structure learning-based dimensionality reduction to visualize the immune landscape of CRC. Our results revealed a complex immune landscape that may provide directions for mRNA vaccine treatment of CRC and define appropriate vaccination patients.


EGCG inhibited bladder cancer T24 and 5637 cell proliferation and migration via PI3K/AKT pathway.
Epigallocatechin-3-gallate (EGCG), the bioactive polyphenol in green tea, has been demonstrated to have various biological activities. We previously found that EGCG inhibited SW780 tumor growth by down-regulation of NF-κB and MMP-9. This study demonstrated that EGCG inhibited bladder cancer T24 and 5637 cell proliferation and migration via PI3K/AKT pathway, without modulation of NF-κB. Our results showed that treatment of EGCG resulted in significant inhibition of cell proliferation by induction of apoptosis, without obvious toxicity to normal bladder SV-HUC-1 cells. EGCG also inhibited 5637 and T24 cell migration and invasion at 25-100 μM. Western blot confirmed that EGCG induced apoptosis in T24 and 5637cells by activation of caspases-3 and PARP. Besides, EGCG up-regulated PTEN and decreased the expression of phosphorylated PI3K, AKT in both T24 and 5637 cells. In addition, animal study demonstrated that EGCG (100 mg/kg, i.p. injected daily for 4 weeks) decreased the tumor weight in mice bearing T24 tumors by 51.2%, as compared with the untreated control. EGCG also decreased the expression of phosphorylated PI3K and AKT in tumor, indicating the important role of PI3K/AKT in EGCG inhibited tumor growth. When AKT was inhibited, EGCG showed no obvious effect in cell migration in T24 and 5637 cells. In conclusion, our study elucidated that EGCG was effective in inhibition of T24 and 5637 cell proliferation and migration, and presented evidence that EGCG inhibited cell proliferation and tumor growth by modulation of PI3K/AKT pathway.


Clinical and hematological response to hydroxyurea in a patient with Hb Lepore/beta-thalassemia.
The possibility of increasing Hb F in vivo using drugs like 5-azacytidine, hydroxyurea, and butyrate has been established. However, in many cases this does not entail an increase in total hemoglobin. We report on a patient with Hb Lepore/beta-thalassemia being treated with hydroxyurea (30 mg/Kg/day) because of the presence of erythroid extramedullary masses with severe neurological abnormalities. During therapy the patient showed a remarkable improvement in neurological signs due to the reduction in extra-medullary masses, a significant increase in both total hemoglobin (from 5.8 to 9.7 g/dl) and Hb F (from 4.9 g/dl to 9.1 g/dl). The marked improvement in hemoglobin level in our patient with Hb Lepore/beta-thalassemia suggests gamma-globin gene activation due to the DNA structure determined by the crossover event.


Differences and inequalities in relation to access to renal replacement therapy in the BRICS countries.
End-stage renal disease (ESRD) is an important public health problem, especially in developing countries due to the high level of economic resources needed to maintain patients in the different programs that make up renal replacement therapy (RRT). To analyze the differences and inequalities involved in access to RRT in the BRICS countries (Brazil, Russian Federation, India, China and South Africa). This is an applied, descriptive, cross-sectional, quantitative and qualitative study, with documentary analysis and a literature review. The sources of data were from national censuses and scientific publications regarding access to RRT in the BRICS countries. There is unequal access to RRT in all the BRICS countries, as well as the absence of information regarding dialysis and transplants (India), the absence of effective legislation to inhibit the trafficking of organs (India and South Africa) and the use of deceased prisoners as donors for renal transplants (China). The construction of mechanisms to promote the sharing of benefits and solidarity in the field of international cooperation in the area of renal health involves the recognition of bioethical issues related to access to RRT in the BRICS countries.


Remaining Gap in HIV Testing Uptake Among Female Sex Workers in Iran.
We estimated the prevalence of recent HIV testing (i.e., having an HIV test during the last 12 months and knew the results) among 1295 HIV-negative Iranian female sex workers (FSW) in 2015. Overall, 70.4% (95% confidence intervals: 59.6, 79.3) of the participants reported a recent HIV testing. Concerns about their HIV status (83.2%) was reported as the most common reason for HIV testing. Incarceration history, having >5 paying partners, having >1 non-paying partner, receiving harm reduction services, utilizing healthcare services, and knowing an HIV testing site were significantly associated with recent HIV testing. In contrast, outreach participants, having one non-paying sexual partner, and self-reported inconsistent condom use reduced the likelihood of recent HIV testing. HIV testing uptake showed a ~2.5 times increase among FSW since 2010. While these findings are promising and show improvement over a short period, HIV testing programs should be expanded particularly through mobile and outreach efforts.


Voltammetric biosensor for coronavirus spike protein using magnetic bead and screen-printed electrode for point-of-care diagnostics.
The rapid spread of the novel human coronavirus 2019 (COVID-19) and its morbidity have created an urgent need for rapid and sensitive diagnostics. The real-time polymerase chain reaction is the gold standard for detecting the coronavirus in various types of biological specimens. However, this technique is time consuming, labor intensive, and expensive. Screen-printed electrodes (SPEs) can be used as point-of-care devices because of their low cost, sensitivity, selectivity, and ability to be miniaturized. The ability to detect the spike protein of COVID-19 in serum, urine, and saliva was developed using SPE aided by magnetic beads (MBs) and a portable potentiostat. The antibody-peroxidase-loaded MBs were the captured and catalytic units for the electrochemical assays. The MBs enable simple washing and homogenous deposition on the working electrode using a magnet. The assembly of the immunological MBs and the electrochemical system increases the measuring sensitivity and speed. The physical and electrochemical properties of the layer-by-layer modified MBs were systematically characterized. The performance of these immunosensors was evaluated using spike protein in the range 3.12-200 ng mL-1. We achieved a limit of detection of 0.20, 0.31, and 0.54 ng mL-1 in human saliva, urine, and serum, respectively. A facile electrochemical method to detect COVID-19 spike protein was developed for quick point-of-care testing.


Free energy, energy, and entropy of swelling in Cs-, Na-, and Sr-montmorillonite clays.
A Monte Carlo method for grand canonical and grand isoshear ensemble simulations has been used to characterize the free energy, energy, and entropy of clay mineral swelling. The Monte Carlo approach was found to be more efficient at simulating water content fluctuations in the highly constrained clay environment than a previously developed molecular dynamics method. Swelling thermodynamics calculated for Cs-, Na-, and Sr-montmorillonite clays indicate a strong dependence of swelling on the interlayer ion identity, in agreement with various experimental measurements. The Sr clay swells most readily, and both the Na and Sr clays prefer expanded states (two-layer hydrate or greater) when in contact with bulk water. In contrast, swelling is inhibited in the Cs clay. Differences in swelling behavior are traced directly to the tendency of the different ions to hydrate. The swelling free energies are decomposed into their energetic and entropic components, revealing an overall energetic driving force for the swelling phenomena. Entropic effects provide a smaller, mediating role in the swelling processes. The results provide a unique molecular perspective on experimentally well-characterized swelling phenomena.


Impact of naturally occurring variation in the human papillomavirus (HPV) 33 capsid proteins on recognition by vaccine-induced cross-neutralizing antibodies.
We investigated naturally occurring variation within the major (L1) and minor (L2) capsid proteins of human papillomavirus (HPV) genotype 33. Pseudoviruses (PsV) representing HPV33 lineages A1, A2, A3, B and C exhibited comparable particle-to-infectivity ratios and morphology but demonstrated a decreased sensitivity (A2, A3, B and C) to cross-neutralization by HPV vaccine antibodies compared to the A1 sublineage. Chimeric PsVs demonstrated that these differences in sensitivity were due to polymorphisms in the L1 protein, with little or no influence from variation within the L2 protein. Site-directed mutagenesis of the L1 gene identified the DE loop residue 133 and the FG residue 266 as being critical for conferring this differential sensitivity. The use of HPV33 homology models based upon the HPV16 crystal structure suggested that they are likely to act independently on more than one antibody epitope. These data improve our understanding of the potential impact of natural capsid variation on recognition by vaccine antibodies.


Role of second-largest RNA polymerase I subunit Zn-binding domain in enzyme assembly.
The second-largest subunits of eukaryal RNA polymerases are similar to the beta subunits of prokaryal RNA polymerases throughout much of their lengths. The second-largest subunits from eukaryal RNA polymerases contain a four-cysteine Zn-binding domain at their C termini. The domain is also present in archaeal homologs but is absent from prokaryal homologs. Here, we investigated the role of the C-terminal Zn-binding domain of Rpa135, the second-largest subunit of yeast RNA polymerase I. Analysis of nonfunctional Rpa135 mutants indicated that the Zn-binding domain is required for recruitment of the largest subunit, Rpa190, into the RNA polymerase I complex. Curiously, the essential function of the Rpa135 Zn-binding domain is not related to Zn(2+) binding per se, since replacement of only one of the four cysteine residues with alanine led to the loss of Rpa135 function. Even more strikingly, replacement of all four cysteines with alanines resulted in functional Rpa135.


Regulation of the immune response. II. Qualitative and quantitative differences between thymus- and bone marrow-derived lymphocytes in the recognition of antigen.
The specificity of antigen recognition by thymus-derived helper cells (T cells) and antibody was examined in mice, heterologous erythrocyte antigens from sheep (SRBC), goat (GRBC), burro (BRBC), chicken (CRBC), and toad (TRBC) being used. Antibody specificity was tested by a number of functional assays: hemagglutination, hemolysis, and immune suppression. The specificity of T cells was determined by titrating their ability to help the in vitro antitrinitrophenol (TNP) responses of mouse spleen cultures immunized with the hapten coupled to the various test erythrocytes as carrier. Anti-SRBC antibody cross-reacted with GRBC, but not with BRBC, CRBC, or TRBC. In contrast, SRBC-primed helper T cells cross-reacted with both GRBC and BRBC, but not with CRBC or TRBC, indicating a difference in the specificity of antigen recognition between the cellular and the humoral immune responses.


Echocardiographic features of peripartum cardiac failure: the Zaria syndrome.
Zaria has the highest incidence of peripartum cardiac failure, a syndrome of unsettled aetiology. Over the years it has been noted that there has been a decrease in the period of treatment by traditional methods from an average of 90 days to 30 days and with a complete avoidance of hot bed treatment. Twenty-one subjects with the syndrome were studied: they exhibited the same clinical findings as evinced in previous reports from our centre. Echocardiography findings of dilated heart with depressed left ventricular function were noted. There was a statistically significant relationship between the duration of practice and ejection fraction (EF), r=0.36 and P<0.05, duration of practice and left ventricular internal diameter in diastole (LVIDd), r=0.28, P<0.05. Were these subjects suffering from a quiescent heart muscle disease from a previous viral myocarditis and did they require an expanded volume as a trigger? Or are they intolerant of salt requiring only a small increase from the traditional method of treatment to precipitate cardiac failure? Further investigation in the field of molecular biology is required.


Comparative investigation of the genomic regions involved in antigenic variation of the TprK antigen among treponemal species, subspecies, and strains.
Although the three Treponema pallidum subspecies (T. pallidum subsp. pallidum, T. pallidum subsp. pertenue, and T. pallidum subsp. endemicum), Treponema paraluiscuniculi, and the unclassified Fribourg-Blanc treponeme cause clinically distinct diseases, these pathogens are genetically and antigenically highly related and are able to cause persistent infection. Recent evidence suggests that the putative surface-exposed variable antigen TprK plays an important role in both treponemal immune evasion and persistence. tprK heterogeneity is generated by nonreciprocal gene conversion between the tprK expression site and donor sites. Although each of the above-mentioned species and subspecies has a functional tprK antigenic variation system, it is still unclear why the level of expression and the rate at which tprK diversifies during infection can differ significantly among isolates. To identify genomic differences that might affect the generation and expression of TprK variants among these pathogens, we performed comparative sequence analysis of the donor sites, as well as the tprK expression sites, among eight T. pallidum subsp. pallidum isolates (Nichols Gen, Nichols Sea, Chicago, Sea81-4, Dal-1, Street14, UW104, and UW126), three T. pallidum subsp. pertenue isolates (Gauthier, CDC2, and Samoa D), one T. pallidum subsp. endemicum isolate (Iraq B), the unclassified Fribourg-Blanc isolate, and the Cuniculi A strain of T. paraluiscuniculi. Synteny and sequence conservation, as well as deletions and insertions, were found in the regions harboring the donor sites. These data suggest that the tprK recombination system is harbored within dynamic genomic regions and that genomic differences might be an important key to explain discrepancies in generation and expression of tprK variants among these Treponema isolates.


Patient-centered benefit-risk analysis of transcatheter aortic valve replacement.
Background: Aortic stenosis (AS) treatments include surgical aortic valve replacement (SAVR) and transcatheter aortic valve replacement (TAVR). Choosing between SAVR and TAVR requires patients to trade-off several benefits and risks. The objective of this research was to determine which outcomes associated with TAVR and SAVR patients consider most important, collect quantitative data about how patients weigh these benefits and risks, and evaluate patients' preferences for SAVR or TAVR. Methods: Patients with aortic stenosis were recruited from advocacy organization databases. An online adapted swing weighting (ASW) method was used to elicit attribute tradeoffs from 93 patients. The ASW exercise consisted of a series of pairwise comparisons of attributes. Survey data were used to estimate the weight that patients put on the AS treatment attributes, which were incorporated into a quantitative benefit-risk analysis (BRA) to evaluate patients' preferences for TAVR and SAVR. Results: On average, patients put greater value on attributes that favored TAVR than SAVR. The value patients placed on the lower short-term mortality rate, reduced procedural invasiveness, and quicker time to return to normal quality of life associated with TAVR, offset the value they placed on the time over which SAVR has been proven to work. There was substantial heterogeneity in patients' preferences. This was partly explained by age, with differences in preference observed between patients <60 years to those ≥60 years. A Monte Carlo Simulation found that 75.1% of patients prefer TAVR. Conclusions: Most AS patients are willing to tolerate sizable increases in clinical risk in exchange for the benefits of TAVR, resulting in a large proportion of patients preferring TAVR to SAVR. Further work should be undertaken to characterize the heterogeneity in preferences for AS treatment attributes. Shared decision-making tools based on attributes important to patients can support patients' selection of the procedure that best meets their needs.


Improving the quality of electron tomography image volumes using pre-reconstruction filtering.
Electron tomography produces highly magnified 3D image volumes useful for investigating the structure and function of cellular components. Image quality is degraded by multiple scattering events and quantum noise, which depend on the angle at which individual tilt projections are collected. We have adapted a biomedical imaging approach to improve image quality by enhancing individual tilt projections prior to volumetric reconstruction. Specifically, we have developed a family of non-linear anisotropic diffusion (NAD) filters parameterized by the tilt angle. We give a quantitative and qualitative evaluation of our pre-processing approach and the NAD filter. We show an improvement in the reconstructed volumes for tomograms generated from both plastic-embedded and cryo-stabilized samples of malaria parasite-infected erythrocytes.


Optimal cut points of plasma and urine neutrophil gelatinase-associated lipocalin for the prediction of acute kidney injury among critically ill adults: retrospective determination and clinical validation of a prospective multicentre study.
To determine the optimal threshold of blood and urine neutrophil gelatinase-associated lipocalin (NGAL) to predict moderate to severe acute kidney injury (AKI) and persistent moderate to severe AKI lasting at least 48 consecutive hours, as defined by an adjudication panel. A multicentre prospective observational study enrolled intensive care unit (ICU) patients and recorded daily ethylenediaminetetraacetic acid (EDTA) plasma, heparin plasma and urine NGAL. We used natural log-transformed NGAL in a logistic regression model to predict stage 2/3 AKI (defined by Kidney Disease International Global Organization). We performed the same analysis using the NGAL value at the start of persistent stage 2/3 AKI. Of 245 subjects, 33 (13.5%) developed stage 2/3 AKI and 25 (10.2%) developed persistent stage 2/3 AKI. Predicting stage 2/3 AKI revealed the optimal NGAL cutoffs in EDTA plasma (142.0 ng/mL), heparin plasma (148.3 ng/mL) and urine (78.0 ng/mL) and yielded the following decision statistics: sensitivity (SN)=78.8%, specificity (SP)=73.0%, positive predictive value (PPV)=31.3%, negative predictive value (NPV)=95.7%, diagnostic accuracy (DA)=73.8% (EDTA plasma); SN=72.7%, SP=73.8%, PPV=30.4%, NPV=94.5%, DA=73.7% (heparin plasma); SN=69.7%, SP=76.8%, PPV=32.9%, NPV=94%, DA=75.8% (urine). The optimal NGAL cutoffs to predict persistent stage 2/3 AKI were similar: 148.3 ng/mL (EDTA plasma), 169.6 ng/mL (heparin plasma) and 79.0 ng/mL (urine) yielding: SN=84.0%, SP=73.5%, PPV=26.6%, NPV=97.6, DA=74.6% (EDTA plasma), SN=84%, SP=76.1%, PPV=26.8%, NPV=96.5%, DA=76.1% (heparin plasma) and SN=75%, SP=75.8%, PPV=26.1, NPV=96.4%, DA=75.7% (urine). Blood and urine NGAL predicted stage 2/3 AKI, as well as persistent 2/3 AKI in the ICU with acceptable decision statistics using a single cut point in each type of specimen.


Genetic structure of major depression symptoms across clinical and community cohorts.
Diagnostic criteria for major depressive disorder allow for heterogeneous symptom profiles but genetic analysis of major depressive symptoms has the potential to identify clinical and aetiological subtypes. There are several challenges to integrating symptom data from genetically-informative cohorts, such as sample size differences between clinical and community cohorts and various patterns of missing data. We conducted genome-wide association studies of major depressive symptoms in three clinical cohorts that were enriched for affected participants (Psychiatric Genomics Consortium, Australian Genetics of Depression Study, Generation Scotland) and three community cohorts (Avon Longitudinal Study of Parents and Children, Estonian Biobank, and UK Biobank). We fit a series of confirmatory factor models with factors that accounted for how symptom data was sampled and then compared alternative models with different symptom factors. The best fitting model had a distinct factor for Appetite/Weight symptoms and an additional measurement factor that accounted for missing data patterns in the community cohorts (use of Depression and Anhedonia as gating symptoms). The results show the importance of assessing the directionality of symptoms (such as hypersomnia versus insomnia) and of accounting for study and measurement design when meta-analysing genetic association data.


Management of acute procedural pain in the neonatal intensive care unit (NICU).
Neonates experience pain and may be more sensitive to it. Our objective was to assess pain management strategies in Irish neonatal units. We performed a cross-sectional telephone survey of all twenty neonatal units in the Republic of Ireland. Nurse managers or the on call registrar was questioned about their unit's analgesia practices. Four units (20%) had guidelines for neonatal procedural pain management Two units (10%) used the PIPP pain assessment tool. Non-nutritive sucking and swaddling were most popular for minor procedures, but use was infrequent. Eight units (40%) had access to sucrose but use was low in most units. Three units used breast-feeding (15%) for heal lancing, IM injections and venepuncture. Most units (60%) stated emergency situations as the main reason for not using analgesia. Despite growing evidence supporting neonatal pain experience and increased sensitivity, neonates are not often afforded the benefits of intervention in Irish NICUs.


Treatment with the platelet-activating factor antagonist TCV-309 in patients with severe systemic inflammatory response syndrome: a prospective, multi-center, double-blind, randomized phase II trial.
In a prospective randomized, double-blind, placebo-controlled clinical study, the safety and efficacy of the platelet-activating factor antagonist TCV-309 in the treatment of systemic inflammatory response syndrome was studied. In total 29 patients were treated with 1.0 mg/kg TCV-309 twice daily during 7 days or with placebo. Study parameters were as follows: adverse events, 28 and 56 day all cause mortality, multi-organ failure scores, and the inflammatory mediators tumor necrosis factor, interleukin 6, interleukin 8, and soluble E-selectin. There was no difference in number and severity of adverse events between TCV-309- and placebo-treated patients. Day 28 and day 56 mortality was similar in both groups (day 56: 7/12 TCV-309 vs. 9/16 placebo, NS). Pulmonary and hematological failure scores improved significantly in TCV-309-treated patients (p < .05). There was no difference in inflammatory mediator levels between TCV-309- and placebo-treated patients. Treatment with TCV-309 appears to be safe in patients with systemic inflammatory response syndrome and does improve organ failure significantly.


Ricinus communis cyclophilin: functional characterisation of a sieve tube protein involved in protein folding.
The phloem translocation stream of the angiosperms contains a special population of proteins and RNA molecules which appear to be produced in the companion cells prior to being transported into the sieve tube system through the interconnecting plasmodesmata. During this process, these non-cell-autonomous proteins are thought to undergo partial unfolding. Recent mass spectroscopy studies identified peptidyl-prolyl cis-trans isomerase (PPIases) as potential molecular chaperones functioning in the phloem translocation stream (Giavalisco et al. 2006). In the present study, we describe the cloning and characterisation of a castor bean phloem cyclophilin, RcCYP1 that has high peptidyl-prolyl cis-trans isomerase activity. Equivalent enzymatic activity was detected with phloem sap or purified recombinant (His)(6)-tagged RcCYP1. Mass spectrometry analysis of proteolytic peptides, derived from a 22 kDa band in HPLC-fractionated phloem sap, immunolocalisation studies and Western analysis of proteins extracted from castor bean tissues/organs indicated that RcCYP1 is an abundant protein in the companion cell-sieve element complex. Microinjection experiments established that purified recombinant (His)(6)-RcCYP1 can interact with plasmodesmata to both induce an increase in size exclusion limit and mediate its own cell-to-cell trafficking. Collectively, these findings support the hypothesis that RcCYP1 plays a role in the refolding of non-cell-autonomous proteins after their entry into the phloem translocation stream.


Genetic analysis of Saccharomyces cerevisiae H2A serine 129 mutant suggests a functional relationship between H2A and the sister-chromatid cohesion partners Csm3-Tof1 for the repair of topoisomerase I-induced DNA damage.
Collision between a topoisomerase I-DNA intermediate and an advancing replication fork represents a unique form of replicative damage. We have shown previously that yeast H2A serine 129 is involved in the recovery from this type of damage. We now report that efficient repair also requires proteins involved in chromatid cohesion: Csm3; Tof1; Mrc1, and Dcc1. Epistasis analysis defined several pathways involving these proteins. Csm3 and Tof1 function in a same pathway and downstream of H2A. In addition, the pathway involving H2A/Csm3/Tof1 is distinct from the pathways involving the Ctf8/Ctf18/Dcc1 complex, the Rad9 pathway, and another involving Mrc1. Our genetic studies suggest a role for H2A serine 129 in the establishment of specialized cohesion structure necessary for the normal repair of topoisomerase I-induced DNA damage.


In situ hybridization and immunohistochemistry of bone sialoprotein and secreted phosphoprotein 1 (osteopontin) in the developing mouse mandibular condylar cartilage compared with limb bud cartilage.
Mandibular condylar cartilage is often classified as a secondary cartilage, differing from the primary cartilaginous skeleton in its rapid progress from progenitor cells to hypertrophic chondrocytes. In this study we used in situ hybridization and immunohistochemistry to investigate whether the formation of primary (tibial) and secondary (condylar) cartilage also differs with respect to the expression of two major non-collagenous glycoproteins of bone matrix, bone sialoprotein (BSP) and secreted phosphoprotein 1 (Spp1, osteopontin). The mRNAs for both molecules were never expressed until hypertrophic chondrocytes appeared. In the tibial cartilage, hypertrophic chondrocytes first appeared at E14 and the expression of BSP and Spp1 mRNAs was detected in the lower hypertrophic cell zone, but the expression of BSP mRNA was very weak. In the condylar cartilage, hypertrophic chondrocytes appeared at E15 as soon as cartilage tissue appeared. The mRNAs for both molecules were expressed in the newly formed condylar cartilage, although the proteins were not detected by immunostaining; BSP mRNA in the condylar cartilage was more extensively expressed than that in the tibial cartilage at the corresponding stage (first appearance of hypertrophic cell zone). Endochondral bone formation started at E15 in the tibial cartilage and at E16 in the condylar cartilage. At this stage (first appearance of endochondral bone formation), BSP mRNA was also more extensively expressed in the condylar cartilage than in the tibial cartilage. The hypertrophic cell zone in the condylar cartilage rapidly extended during E15-16. These results indicate that the formation process of the mandibular condylar cartilage differs from that of limb bud cartilage with respect to the extensive expression of BSP mRNA and the rapid extension of the hypertrophic cell zone at early stages of cartilage formation. Furthermore, these results support the hypothesis that, in vivo, BSP promotes the initiation of mineralization.


The use of the bi-factor model to test the uni-dimensionality of a battery of reasoning tests.
The Battery of Reasoning Tests 5 (BPR-5) aims to assess the reasoning ability of individuals, using sub-tests with different formats and contents that require basic processes of inductive and deductive reasoning for their resolution. The BPR has three sequential forms: BPR-5i (for children from first to fifth grade), BPR-5 - Form A (for children from sixth to eighth grade) and BPR-5 - form B (for high school and undergraduate students). The present study analysed 412 questionnaires concerning BPR-5i, 603 questionnaires concerning BPR-5 - Form A and 1748 questionnaires concerning BPR-5 - Form B. The main goal was to test the uni-dimensionality of the battery and its tests in relation to items using the bi-factor model. Results suggest that the g factor loadings (extracted by the uni-dimensional model) do not change when the data is adjusted for a more flexible multi-factor model (bi-factor model). A general reasoning factor underlying different contents items is supported.


Black necrosis of the glans penis associated with calciphylaxis: A case report.
Calciphylaxis, known as calcific uremic arteriolopathy, is a rare cause of dry gangrene. Despite an increase in the clinical recognition of demographic characteristics and risk factors associated with calciphylaxis, it remains a poorly understood disease with high mortality. We present a 45-year-old man, who was diagnosed with calciphylaxis disease, with a history of diabetes mellitus, end-stage renal disease and cirrhosis with a half-month evolution of painful dry gangrene on his glans penis and scrotum. The patient also presented with gangrene of fingers. The patient and his family opted for palliative care. However, he died eventually. This case contributed to the current understanding of calciphylaxis. Since no standard treatment is available and the prognosis remained poor, early, and accurate diagnosis of calciphylaxis is important. We here report the current case and provide data for the diagnosis and treatment of this kind of disease.


A Potential Animal Model of Maladaptive Palatable Food Consumption Followed by Delayed Discomfort.
Introduction: Binging is the consumption of larger amounts of food in a briefer period of time than would normally be consumed under similar circumstances. Binging requires palatable food (PF) to trigger abnormal eating, probably reflecting gene × environment interactions. In this study we examined the impact of trait binge eating (BE) and its compulsive nature on the conflict between hedonic eating of PF and anticipation of a delayed aversive effect. We used female rats as an animal model similar to other models of BE. A novel aspect of this model in this paper is the use of a delayed internal aversive effect produced by lactose ingestion. Establishing this model will allow us to better understand the nature of the conflict between immediate reward and its delayed aversive implications. We hypothesized that BE prone (BEP) rats will demonstrate maladaptive decision making, presenting higher motivation toward PF even when this is associated with delayed discomfort. Method: (Phase 1) 52 female adult Wistar rats were divided to two eating profiles: resistant and prone binge eaters (BER/BEP) based on intake of liquid PF (Ensure). Next, all subjects underwent a Lactose Conditioning Protocol (LCP) that included 4 h tests, one baseline and 3 conditioning days (Phase 2), in which solid PF (Oreo cookies) was paired with glucose (control-no internal aversive effect) or lactose, dissolved in liquid PF. Index for PF motivation was PF consumption during the 4 h LCP. To test for memory of lactose conditioning, we performed another LCP with glucose only (anticipation, but no actual lactose-induced discomfort), a week after the last conditioning session. Results: Lactose conditioned BEP showed higher motivation toward PF compared to lactose conditioned BER faced with delayed aversive effects. Only lactose conditioned BER rats devaluated the PF over LCP days, indicating an association between PF and abdominal discomfort. In addition, only lactose conditioned BER presented an adaptive dynamic behavior, by varying PF intake according to consequences. Furthermore, solid PF consumption was predicted by binge size of liquid PF, only for lactose conditioned rats. Conclusions: We established an animal model for a common eating conflict in humans using delayed internal aversive unconditional stimuli.


Effect of chronic administration of PDE5 combined with glycemic control on erectile function in streptozotocin-induced diabetic rats.
Chronic treatment with phosphodiesterase type 5 inhibitors (PDE5) is effective in an animal model of diabetes-induced erectile dysfunction (DMED). In addition, recent research indicates that glycemic control can restore DMED. We evaluated the effect of chronic administration of PDE5 combined with glycemic control on DMED. Sprague-Dawley rats (8 weeks old) were divided into five groups (n = 10 each): normal control (C), diabetes (DM), DM treated with insulin (DM-I), DM treated with PDE5 (DM-P), and DM treated with insulin and PDE5 (DM-I + P). Rats in the diabetic groups received an injection of streptozotocin (45 mg/kg). After 10 weeks of induced diabetes, the DM-I group was treated with a daily injection of neutral protamine Hagedorn, and the DM-P group was treated with a daily dosage of 20 mg/kg PDE5 (DA-8159) for 4 weeks. The DM-I + P group was treated with both treatments simultaneously. After 14 weeks of induced diabetes, an evaluation of erectile function and histological and biochemical markers of corporal tissue was performed. Erectile function and histological and biochemical markers in corporal tissue. Rats in the DM group showed markedly lower erectile parameters than those in the C group, whereas rats in the DM-I and DM-P groups showed intermediate erectile function between the DM and C groups. Rats in the DM-I + P group showed restored erectile function, comparable with group C. A comparison of apoptotic index, expression of the endothelial marker, and phosphorylation of endothelial nitric oxide synthase and Akt displayed a similar pattern with the results from cavernosometry (DM < DM-I = DM-P < DM-I + P = C, P < 0.05). The distribution of phosphorylated myosin phosphatase target subunit 1 was in the reverse order. Chronic administration of PDE5 or glycemic control with insulin resulted in restoration of overt DMED. The combination of both treatments was superior to monotherapy with insulin or PDE5.


Black and White and Shades of Gray [State of the Art].
"Moderation in all things" is a popular saying that many of us have heard all our lives. Still, a good number of people seem to have forgotten the sentiment behind this advice. Instead of looking for the good that exists within the bad and the bad that dwells within the good, people are choosing to line up behind one extreme or another. Nuances are being ignored in favor of strong positions on either side of the middle. This has led inevitably to polarization, partisanship, and balkanization in our society. For some reason, moderation has been forgotten.


The costs and cost effectiveness of providing first-trimester, medical and surgical safe abortion services in KwaZulu-Natal Province, South Africa.
Despite a liberal abortion law, access to safe abortion services in South Africa is challenging for many women. Medication abortion was introduced in 2013, but its reach remains limited. We aimed to estimate the costs and cost effectiveness of providing first-trimester medication abortion and manual vacuum aspiration (MVA) services to inform planning for first-trimester service provision in South Africa and similar settings. We obtained data on service provision and outcomes from an operations research study where medication abortion was introduced alongside existing MVA services in public hospitals in KwaZulu-Natal province. Clinical data were collected through interviews with first-trimester abortion clients and summaries completed by nurses performing the procedures. In parallel, we performed micro-costing at three of the study hospitals. Using a model built in Excel, we estimated the average cost per medical and surgical procedure and determined the cost per complete abortion performed. Results are presented in 2015 US dollars. A total of 1,129 women were eligible for a first trimester abortion at the three study sites. The majority (886, 78.5%) were eligible to choose their abortion procedure; 94.1% (n = 834) chose medication abortion. The total average cost per medication abortion was $63.91 (52.32-75.51). The total average cost per MVA was higher at $69.60 (52.62-86.57); though the cost ranges for the two procedures overlapped. Given average costs, the cost per complete medication abortion was lower than the cost per complete MVA despite three (0.4%) medication abortion women being hospitalized and two (0.3%) having ongoing pregnancies at study exit. Personnel costs were the largest component of the total average cost of both abortion methods. This analysis supports the scale-up of medication abortion alongside existing MVA services in South Africa. Women can be offered a choice of methods, including medication abortion with MVA as a back-up, without increasing costs.


Age-Stratified Lithium Therapeutic Ranges for Older Adults with Bipolar Disorder - from Awareness to an Action Plan.
Lithium is the first-line treatment for maintenance therapy in bipolar disorder. It is an effective mood stabilizer agent, and may have potential benefits in neuroprotection and reducing the risk of suicide. Toxicity has been a concern in recent decades, particularly in older adults (≥60 years). In 2019, the Older Adults Task Force within the International Society for Bipolar Disorder (ISBD) published recommendations for age-stratified lithium therapeutic ranges for therapy of Older Age Bipolar Disorder (OABD), namely 0.4 - 0.8 mmol/L for ages 60 to 79 and 0.4 - 0.7 mmol/L for ages 80 and above. Clinical laboratory practice surveys in Canada indicated that adoption and implementation of the proposed ranges has been limited to date. In this article, we describe the approach and steps taken to evaluate and implement recommended lithium therapeutic ranges in Ontario and other provinces in Canada for laboratory quality improvement. Sources of variation in lithium reporting practices are discussed and shared here to highlight potential barriers to implementation. The overall goal of this article is to bring attention across the global laboratory community that lower lithium therapeutic target ranges in older patients are crucial for patient safety in OABD.


[The pancreolaurinic test in patients with chronic pancreatitis and in patients with perforated acute pancreatitis].
They presented a valuation of exocrine pancreatic function using pancreolaurinic test in 15 patients with chronic pancreatitis and in 34 patients after acute pancreatitis. In all patients with chronic pancreatitis they noted an insufficiency of exocrine pancreatic function with association of disorders of tolerance for carbohydrates during a year after a Puestow's operation. They noted a handicap of exocrine pancreatic function in 42% patients just after acute pancreatitis, in 18% patients a year acute pancreatitis and in 26% patients after 2-6 years from illness with association of disorders of tolerance for carbohydrates.


One-year coronary bypass graft patency: a randomized comparison between off-pump and on-pump surgery angiographic results of the PRAGUE-4 trial.
Off-pump coronary bypass surgery has become a widely used technique during recent years. However, limited data are available with regard to 1-year patency of bypass grafts implanted on the beating heart in unselected consecutive bypass surgery candidates. The aim of this study was to compare 1-year angiographic patency of bypass grafts done on the beating heart (off pump) with those done classically (on pump). The PRAGUE-4 trial randomized 400 consecutive nonselected cardiac surgery candidates into group A (on pump; n=192) and group B (off pump; n=208). One-year follow-up coronary angiography was done in 255 patients. The arterial graft patency after 1 year was 91% in both groups. Saphenous graft patency was 59% (on pump) versus 49% (off pump; P=NS). Saphenous graft patency per patient was lower in the off-pump group: 0.7 patent anastomosis per patient versus 1.1 patent anastomosis in the on-pump group (P<0.01). There were 46% on-pump patients with all grafts patent versus 52% off-pump patients (P=NS). Grafts anastomosed distally to collateralized chronic total occlusions of native coronary arteries remained patent in 100% on the left anterior descending artery compared with 23% on other arteries (P<0.0001). The patency of arterial coronary bypass grafts done on the beating heart is excellent and equal to grafts done on pump. The off-pump procedure in the unselected patient population results in fewer patent saphenous grafts per patient.


Applicability of two automated disintegration apparatuses for rapidly disintegrating (mini)tablets.
Orally disintegrating (mini)tablets (OD(M)Ts) are of interest in the field of pharmaceutics. Their orodispersible character is defined by the disintegration time, which is measured with a basket apparatus according to the European Pharmacopoeia. This method, however, lacks applicability for ODTs and especially ODMTs. New disintegration apparatuses have been described in literature, but a qualification to assess the applicability has not been described. A qualification procedure for two automated disintegration apparatuses, OD-mate and Hermes apparatus, is introduced. Aspects of the operational qualification as well as precision and accuracy regarding a performance qualification were evaluated for both apparatuses analog to the ICH guideline Q2. While the OQ study is performed separately for each apparatus, accuracy and precision were performed following the same protocol for both testers. Small RSDs (16.9% OD-mate; 15.2% Hermes compared to 32.3% for the pharmacopeial method) were found despite very fast disintegration times (1.5 s for both apparatuses). By comparing these RSDs to practical examples, the authors propose threshold values for repeatability depending on the mean disintegration time. Obtained results from the qualification were used to assess the applicability of both apparatuses.


Assembly modulation of PDI derivative as a supramolecular fluorescence switching probe for detection of cationic surfactant and metal ions in aqueous media.
We report an amphiphilic perylene diimide (1), a bimolecular analog of l-3,4-dihydroxyphenylalanine (L-DOPA), as a reversible fluorescence switching probe for the detection and sensing of cationic surfactants and Fe(3+)/Cu(2+) in an aqueous media respectively by means of host-guest interactions driven assembly and disassembly of 1. Photophysical studies of 1, going from dimethyl sulfoxide (DMSO) (State-I) to pure aqueous medium (State-II), suggested the formation of self-assembled aggregates by displaying very weak fluorescence emission along with red shifted broad absorption bands. Interestingly, the cationic surfactant cetyltrimethylammonium bromide (CTAB) could disassemble 1 in miceller conditions by restoring bright yellow fluorescence and vibronically well-defined (Franck-Condon progressions A0-0/A0-1 ≈ 1.6) absorption bands of 1 over other neutral and anionic surfactants (State-III). Owing to the metal chelating nature of L-DOPA, 1 was able to sense Fe(3+) and Cu(2+) among a pool of other metal ions by means of fluorescence switching off state, attributed to metal interaction driven assembly of 1 (State-IV). Such metallosupramolecular assemblies were found to reverse back to the fluorescence switching on state using a metal ion chelator, diethylenetriaminepentaacetic acid (DTPA, State-III), further signifying the role of metal ions toward assembly of 1. Formation of assembly and disassembly could be visualized by the diminished and increased yellow emission under green laser light. Further, the assembly-disassembly modulation of 1 has been extensively characterized using infrared (IR), mass spectrometry, microscopy and dynamic light scattering (DLS) techniques. Therefore, modulation of the molecular self-assembly of PDI derivative 1 in aqueous media (assembled state, State-II) by means of host-guest interactions provided by micellar structures of CTAB (disassembled state, State-III), metal ion (Fe(3+) and Cu(2+)) interactions (assembled state, State-IV) and metal ion sequestration using DTPA (disassembled state, State-III) is viewed as a supramolecular reversible fluorescence switching off-on probe for cationic surfactant CTAB and Fe(3+)/Cu(2+).


Predictive Model for Operative Intervention after Blunt Abdominal Trauma in Children with Equivocal CT Findings: A Pilot Study.
To study the clinical and radiologic factors predicting the need for surgical intervention after blunt abdominal trauma (BAT) in children with equivocal computed tomography (CT) scan findings. We performed a retrospective review of the trauma database at our level I pediatric trauma center between 2011 and 2019. We selected patients with BAT and equivocal findings for surgical intervention on CT scan. We studied five factors: abdominal wall bruising (AWB), abdominal pain/tenderness (APT), thoracolumbar fracture, the presence of free fluid (FF), and the presence of solid organ injury, all previously reported in the literature to predict the need for operative intervention. We used t-test, the Kruskal-Wallis test and logistic regression to study the association of these factors with the need for operation in our pediatric cohort. Of 3044 blunt trauma patients, 288 had abdominal CT scans with 61 patients demonstrating equivocal findings. Operation was performed for 12 patients (19.7%) confirming surgically correctable traumatic injuries. The need for surgical intervention was significantly associated with the age of the patients (P = 0.03), the presence of APT (P = 0.001), AWB (P = 0.01), and FF (P = 0.04). The presence of thoracolumbar fracture and solid organ injury were not significantly associated with the need for operation. For the subset of 37 patients who were injured in a motor vehicle crash, five (13.5%) required surgical intervention, which was significantly associated with the presence of AWB (P = 0.04), APT (P = 0.01), and FF (P = 0.03). A predictive model that used these factors produced a receiver operating characteristic curve of 0.86. In cases of equivocal abdominal CT scan findings to evaluate BAT in children, the presence of abdominal wall tenderness, AWB, or FF may be significant factors predicting more accurately the need for operative intervention. A predictive model using the combination of clinical and image findings might determine with more certainty, the need for surgical intervention in children with BAT and equivocal CT findings. Validation on a larger multi-institutional data set should be done.


[Nephrogenic systemic fibrosis].
Nephrogenic systemic fibrosis (NFS) is a relatively newly discovered disease, which has been reported in patients with renal failure. NSF usually develops after exposure to Gadolinium (Gd) based contrast media, which are used in magnetic resonance (MR) studies. Nephrogenic systemic fibrosis has both cutaneous and deep, visceral manifestations, involving muscles, lungs, heart and more. The course of NFS is usually progressive, with no effective treatment reported, except for renal function improvement. Awareness to NFS is important not only for early diagnosis of affected patients, but also in preventing new cases. According to current FDA guidelines for imaging patients with renal failure, use of CT with iodinated contrast media is preferred in patients with renal failure, over MR studies with Gd in patients with renal failure.


Seasonal variation of acute toxoplasmic lymphadenopathy in the United States.
We describe the seasonal variation of acute toxoplasmosis in the United States. Acute toxoplasmic lymphadenopathy (ATL) can be a surrogate of acute toxoplasmosis in patients in whom the date of onset of lymphadenopathy matches the window of acute infection predicted by serological tests performed at a reference laboratory. We used the electronic database of the Palo Alto Medical Foundation Toxoplasma Serology Laboratory (PAMF-TSL) (1997-2011) to identify cases of ATL. We tested the uniformity of distribution of ATL cases per month, across the 12 calendar months, using circular statistics uniformity tests. We identified 112 consecutive cases of ATL. The distribution of cases was not uniform across the 12 calendar months. We observed the highest peak of cases in December and a second highest peak in September. Similar months were identified in patients with acute toxoplasmosis in rural areas in France. The results were similar when we performed weighted analyses, weighting for the total number of Toxoplasma gondii IgG tests performed per month in the PAMF-TSL laboratory. This is the largest study to date of the seasonal variation of ATL in the United States. Physicians should advise high-risk individuals to avoid risk factors associated with T. gondii infections especially around those months.


Tighter precision target required for lactate testing in patients with lactic acidosis.
Allowable analytic errors are generally based on biologic variation in normal, healthy subjects. Some analytes like blood lactate have low concentrations in healthy individuals and resultant allowable variation is large when expressed as a coefficient of variation (CV). In Ricós' compendium of biologic variation, the relative pooled intra-individual lactate variation (si) averages 27% and the desirable imprecision becomes 13.5%. We derived biologic variability (sb) from consecutive patient data and demonstrate that sb of lactate is significantly lower. A data repository provided lactate results measured over 18 months in the General Systems intensive care unit (ICU) at the University of Alberta Hospital in Edmonton, Canada. In total 54,000 lactate measurements were made on two point-of-care Radiometer 800 blood gas systems operated by Respiratory Therapy. The standard deviations of duplicates (SDD) were tabulated for the intra-patient lactates that were separated by 0-1, 1-2...up to 16 h. The graphs of SDD vs. time interval were approximately linear; the y-intercept provided by the linear regression represents the sum of sb and short-term analytic variation (sa):y₀=(sa²+sb²)½. The short-term sa was determined from imprecisions provided by Radiometer and confirmed with onsite controls. The derivation of sb was performed for multiple patient ranges of lactate. The relative desirable lactate imprecision for patients with lactic acidosis is about half that of normal individuals. As such, evaluations of lactate measurements must use tighter allowable error limits.


Binding Debye-Hückel theory for associative electrolyte solutions.
This study presents a new equation of state (EOS) for charged hard sphere fluids that incorporates ion-ion association. The EOS is developed using the Debye-Hückel (DH) theory, reference cavity approximation, and Wertheim's theory. Predictive accuracy is evaluated by comparing the model's predictions with Monte Carlo simulations for various charged hard-sphere fluids. The assessment focuses on mean ionic activity coefficient, individual ionic activity coefficient, and osmotic coefficients. The results demonstrate good agreement between the model and simulations, indicating its success for different electrolyte systems. Incorporating ion-ion association improves accuracy compared to the DH theory. The importance of the cavity function and ion-dipole interactions is emphasized in accurately representing structural properties. Overall, the developed EOS shows promising predictive capabilities for charged hard sphere fluids, providing validation and highlighting the significance of ion-ion association in thermodynamic predictions of electrolyte solutions.


Genomic analysis of Staphylococcus aureus from the West African Dwarf (WAD) goat in Nigeria.
Staphylococcus aureus can colonize various host species, and human-animal interaction is a significant factor for cross-species transmission. However, data on S. aureus colonization in animals, particularly on ruminants in close contact with humans, is limited. The West African Dwarf (WAD) goat is among the earliest domesticated ruminant associated with rural dwellers and small-holder farmers in sub-Saharan Africa. This study aimed to investigate the population structure, antibiotic resistance, and virulence gene determinants of S. aureus from the WAD goat in Nigeria. Nasal samples were obtained from the WAD goat in five markets in Osun State, South-West Nigeria. S. aureus was characterized by antibiotic susceptibility testing, detection of virulence determinants, spa typing, and multilocus sequence typing (MLST). Representative isolates were selected for whole-genome sequencing, biofilm, and cytotoxicity assay. Of the 726 nasal samples obtained from the WAD goat, 90 S. aureus (12.4%) were recovered. Overall, 86 isolates were methicillin-susceptible, and four were mecA-positive (i.e., methicillin-resistant S. aureus [MRSA]). A diverse S. aureus clonal population was observed (20 sequence types [STs] and 37 spa types), while 35% (13/37) and 40% (8/20) were new spa types and STs, respectively. Eleven MLST clonal complexes (CC) were identified (CC1, CC5, CC8, CC15, CC30, CC45, CC97, CC121, CC133, CC152, CC522). The MRSA isolates were designated as t127-ST852-CC1-SCCmec type VII, t4690-ST152-CC152-SCCmec type Vc, and t8821-ST152-CC152-SCCmec type Vc. Phylogenetic analysis revealed that 60% (54/90) of all isolates were associated with ruminant lineages (i.e., CC133, CC522). Panton-Valentine Leukocidin (PVL)-positive S. aureus was identified in CC1, CC30, CC121, and CC152. For the CC522 isolates, we illustrate their pathogenic potential by the detection of the toxic shock syndrome gene and hemolysins, as well as their strong cytotoxicity and ability to form biofilms. This is the first detailed investigation on the genomic content of S. aureus from the WAD goat in Nigeria. The S. aureus population of the WAD goat consists mainly of ruminant-associated lineages (e.g., CC133, CC522), interspersed with human-associated clones, including PVL-positive MRSA CC1 and CC152.


[Listeria monocytogenes in pasteurized milk].
An haemolytic Listeria monocytogenes strain pathogenic to mice was isolated from 6 out of 28 (21.4%) pasteurized milk samples (3.2% fat milk treated at 78 degrees C for 15 s) marketed by a Madrid processing plant. Listeria grayi was recovered from 25 of the samples (89.2%) and L. innocua from 3 samples (10.7%). One milk sample was contaminated with L. welshimeri. No strains of L. ivanovii, L. seeligeri, L. murrayi, or L. denitrificans were isolated. These results show that pathogenic Listeria strains can be isolated from pasteurized milk and reinforce the hypothesis that this food product may be the source of numerous human listeriosis.


Characterization of two aerobic ultramicrobacteria isolated from urban soil and a description of Oxalicibacterium solurbis sp. nov.
Two strains of aerobic, non-spore-forming, Gram-negative, rod-shaped bacteria (ND5 and MY14(T)), previously isolated from urban soil using the membrane-filter enrichment technique, were characterized. Analysis of their 16S rRNA gene sequence grouped strains ND5 and MY14(T) within the family Oxalobacteraceae (Betaproteobacteria). The highest pairwise sequence similarities for strain ND5 were found with members of the genus Herminiimonas, namely with Herminiimonas saxobsidens NS11(T) (99.8%) and Herminiimonas glaciei UMB49(T) (99.6%). Although some fatty acid profiles, physiological and biochemical differences exist between strain ND5 and the respective Herminiimonas-type strains, DNA-DNA hybridization experiments confirm that strain ND5 is a member of the H. glaciei genospecies. Taxonomical analyses revealed a wider range of variability within this genus than considered previously. The highest pairwise nucleotide similarity for strain MY14(T) was found with Oxalicibacterium flavum (96.8%). Phylogenetic analyses based on 16S rRNA and cpn60 gene sequences, DNA-DNA hybridization, fatty acid profiles, physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain MY14(T) from other Oxalicibacterium species representing a new species, for which the name Oxalicibacterium solurbis sp. nov. (type strain MY14(T)=NBRC 102665(T),=CCM 7664(T)) is proposed.


An estimate of the economic burden and premature deaths due to vitamin D deficiency in Canada.
The objective of this work is to estimate the economic burden and premature death rate in Canada attributable to low serum 25-hydroxyvitamin D (25(OH)D) levels. Vitamin D deficiency has been linked to many diseases and conditions in addition to bone diseases, including many types of cancer, several bacterial and viral infections, autoimmune diseases, cardiovascular diseases, and adverse pregnancy outcomes. Canadians have mean serum 25(OH)D levels averaging 67 nmol/L. The journal literature was searched for papers reporting dose-response relationships for vitamin D indices and disease outcomes. The types of studies useful in this regard include randomized controlled trials, observational, cross-sectional, and ecological studies, and meta-analyses. The mortality rates for 2005 were obtained from Statistics Canada. The economic burden data were obtained from Health Canada. The estimated benefits in disease reduction were based on increasing the mean serum 25(OH)D level to 105 nmol/L. It is estimated that the death rate could fall by 37,000 deaths (22,300-52,300 deaths), representing 16.1% (9.7-22.7%) of annuals deaths and the economic burden by 6.9% (3.8-10.0%) or $14.4 billion ($8.0 billion-$20.1 billion) less the cost of the program. It is recommended that Canadian health policy leaders consider measures to increase serum 25(OH)D levels for all Canadians.


Identification of comS, a gene of the srfA operon that regulates the establishment of genetic competence in Bacillus subtilis.
Genetic competence (the ability to internalize exogenous DNA) in Bacillus subtilis is dependent on a regulatory pathway that activates the expression of a battery of competence-specific genes. The srfA operon, encoding the subunits of surfactin synthetase, which catalyzes the nonribosomal synthesis of the peptide antibiotic surfactin, also functions in the competence regulatory pathway. The DNA encoding only one of the seven amino acid-activating domains of surfactin synthetase, the valine-activating domain (srfAB1), is necessary for competence. Deletion analysis revealed that a 569-bp fragment of srfAB1, fused to the srfA promoter, complements a srfA deletion mutation (delta srfA) with respect to competence. This fragment contains an open reading frame consisting of 46 amino acids (orf46), which is out of frame with srfAB1. A frameshift mutation in srfAB upstream of orf46 has no effect on competence but a frameshift and nonsense mutation in orf46 resulted in failure to complement the delt srfA mutation. These results indicate that orf46 encodes the srfA-associated competence regulatory factor. Computer-aided analysis of the putative orf46 product (ComS) shows similarity to the homeodomain of the POU domain class of eukaryotic transcriptional regulators.


Enabling low cost biopharmaceuticals: high level interferon alpha-2b production in Trichoderma reesei.
The filamentous fungus Trichoderma reesei has tremendous capability to secrete over 100 g/L of proteins and therefore it would make an excellent host system for production of high levels of therapeutic proteins at low cost. We have developed T. reesei strains suitable for production of therapeutic proteins by reducing the secreted protease activity. Protease activity has been the major hindrance to achieving high production levels. We have constructed a series of interferon alpha-2b (IFNα-2b) production strains with 9 protease deletions to gain knowledge for further strain development. We have identified two protease deletions that dramatically improved the production levels. Deletion of the subtilisin protease slp7 and the metalloprotease amp2 has enabled production levels of IFNα-2b up to 2.1 and 2.4 g/L, respectively. With addition of soybean trypsin protease inhibitor the level of production improved to 4.5 g/L, with an additional 1.8 g/L still bound to the secretion carrier protein. High levels of IFNα-2b were produced using T. reesei strains with reduced protease secretion. Further strain development can be done to improve the production system by reducing protease activity and improving carrier protein cleavage.


Concentrations and bioconcentration factors of leaf microelements in response to environmental gradients in drylands of China.
Determining response patterns of plant leaf elements to environmental variables would be beneficial in understanding plant adaptive strategies and in predicting ecosystem biogeochemistry processes. Despite the vital role of microelements in life chemistry and ecosystem functioning, little is known about how plant microelement concentrations, especially their bioconcentration factors (BCFs, the ratio of plant to soil concentration of elements), respond to large-scale environmental gradients, such as aridity, soil properties and anthropogenic activities, in drylands. The aim of the present study was to fill this important gap. We determined leaf microelement BCFs by measuring the concentrations of Mn, Fe, Ni, Cu and Zn in soils from 33 sites and leaves of 111 plants from 67 species across the drylands of China. Leaf microelement concentrations were maintained within normal ranges to satisfy the basic requirements of plants, even in nutrient-poor soil. Aridity, soil organic carbon (SOC) and electrical conductivity (EC) had positive effects, while soil pH had a negative effect on leaf microelement concentrations. Except for Fe, aridity affected leaf microelement BCFs negatively and indirectly by increasing soil pH and SOC. Anthropogenic activities and soil clay contents had relatively weak impacts on both leaf microelement concentrations and BCFs. Moreover, leaf microelement concentrations and BCFs shifted with thresholds at 0.89 for aridity and 7.9 and 8.9 for soil pH. Woody plants were positive indicator species and herbaceous plants were mainly negative indicator species of leaf microelement concentrations and BCFs for aridity and soil pH. Our results suggest that increased aridity limits the absorption of microelements by plant leaves and enhances leaf microelement concentrations. The identification of indicator species for the response of plant microelements to aridity and key soil characteristics revealed that woody species in drylands were more tolerant to environmental changes than herbaceous species.


Human RORγt(+)CD34(+) cells are lineage-specified progenitors of group 3 RORγt(+) innate lymphoid cells.
Group 3 innate lymphoid cells (ILC3s) are defined by the expression of the transcription factor RORγt, which is selectively required for their development. The lineage-specified progenitors of ILC3s and their site of development after birth remain undefined. Here we identified a population of human CD34(+) hematopoietic progenitor cells (HPCs) that express RORγt and share a distinct transcriptional signature with ILC3s. RORγt(+)CD34(+) HPCs were located in tonsils and intestinal lamina propria (LP) and selectively differentiated toward ILC3s. In contrast, RORγt(-)CD34(+) HPCs could differentiate to become either ILC3s or natural killer (NK) cells, with differentiation toward ILC3 lineage determined by stem cell factor (SCF) and aryl hydrocarbon receptor (AhR) signaling. Thus, we demonstrate that in humans RORγt(+)CD34(+) cells are lineage-specified progenitors of IL-22(+) ILC3s and propose that tonsils and intestinal LP, which are enriched both in committed precursors and mature ILC3s, might represent preferential sites of ILC3 lineage differentiation.


Human intestinal permeability.
This review focuses on permeability measurements in humans, briefly discussing different perfusion techniques, the relevance of human Peff values, and various aspects of in vivo transport mechanisms. In addition, human Peff values are compared with corresponding data from three preclinical transport models. The regional human jejunal perfusion technique has been validated in several important ways. One of the most important findings is that there is a good correlation between the measured human effective permeability values and the extent of absorption of drugs in humans determined by pharmacokinetic studies. Estimations of the absorption half-lives from the measured Peff agree very well with the time to maximal amount of the dose absorbed achieved after an oral dose in humans. We have also shown that it is possible to determine the Peff for carrier-mediated transported compounds and to classify them according to the proposed biopharmaceutical classification system (BCS). Furthermore, human in vivo permeabilities can be predicted using preclinical permeability models, such as in situ perfusion of rat jejunum, the Caco-2 model, and excised intestinal segments in the Ussing chamber. The permeability of passively transported compounds can be predicted with a particularly high degree of accuracy. However, special care must be taken for drugs with a carrier-mediated transport mechanism, and a scaling factor has to be used. Finally, the data obtained in vivo in humans emphasize the need for more clinical studies investigating the effect of physiological in vivo factors and molecular mechanisms influencing the transport of drugs across the intestinal and as well as other membrane barriers. It will also be important to study the effect of antitransport mechanisms (multidrug resistance, MDR), such as efflux by P-glycoprotein(s) and gut wall metabolism, for example CYP 3A4, on bioavailability.


Neurofunctional effects of developmental sodium fluoride exposure in rats.
Contrasting studies on the toxic effects of sodium fluoride (NaF) during developmental stages of Wistar rats, lead us to investigate the neurofunctional effects caused by its perinatal exposure, devoid of any overt sign of toxicity and/or gross malformation. NaF solution was administered to pregnant rats by intragastric gavage at a daily dose of 2.5 and 5.0 mg/kg from gestational day 0 to day 9 after parturition. Developmental NaF exposure caused sex and dose specific behavioural deficits which affected males more than females in the majority of the evaluated end-points. In particular, the perinatal exposure to NaF 5.0 mg/kg, significantly affected learning, memory, motor coordination and blood pressure only in male rats. Conversely, a lack of habituation upon the second presentation of the objects and failure in the ability to discriminate between the novel and the familiar object were observed only in NaF 5.0 mg/kg female rats. Finally, a significant impairment of sexual behaviour was observed in male rats at both NaF dose levels. The present data indicate that perinatal rat exposure to NaF results in long lasting functional sex-specific alterations which occur at fluoride levels approaching those experienced by offspring of mothers.


Complete factorial design to adjust pH and sugar concentrations in the inoculum phase of Ralstonia solanacearum to optimize P(3HB) production.
Poly(3-hydroxybutyrate) (P(3HB)) is a biodegradable plastic biopolymer that accumulates as lipophilic inclusions in the cytoplasm of some microorganisms. The biotechnological process by which P(3HB) is synthesized occurs in two phases. The first phase involves cell growth in a complex culture medium, while the second phase involves polymer accumulation in the presence of excess carbon sources. As such, the efficiency of the second phase depends on the first phase. The aim of this study was to evaluate culture media with different concentrations of sucrose and glucose and different pH values in the inoculum phase of Ralstonia solanacearum RS with the intention of identifying methods by which the biomass yield could be increased, subsequently enhancing the yield of P(3HB). The culture medium was formulated according to the experimental planning type of central composite rotational design 22. The independent variables were pH and sugar concentration (sucrose and glucose), and the dependent variables were OD600nm, dry cell weight (DCW), and P(3HB) yield. The highest cell growth, estimated by the OD600nm (20.6) and DCW (5.35) values, was obtained when sucrose was used in the culture medium at a concentration above 35 g.L-1 in combination with an acidic pH. High polymer (45%) accumulation was also achieved under these conditions. Using glucose, the best results for OD600nm (12.5) and DCW (2.74) were also obtained at acidic pH but with a sugar concentration at the minimum values evaluated. Due to the significant accumulation of polymer in the cells that were still in the growth phase, the accumulating microorganism P(3HB) Ralstonia solanacearum RS can be classified as having type II metabolism in relation to the polymer accumulation phase, which is different from other Ralstonia spp. studied until this time.


Combinatorial library of chalcogen-containing lipidoids for intracellular delivery of genome-editing proteins.
Protein based therapeutics with high specificities and low off-target effects are used for transient and accurate manipulation of cell functions. However, developing safe and efficient carriers for intracellular delivery of active therapeutic proteins is a long-standing challenge. Here we report a combinatorial library of chalcogen (O, S, Se) containing lipidoid nanoparticles (LNPs) as efficient nanocarriers for intracellular delivery of negatively supercharged Cre recombinase ((-30)GFP-Cre) and anionic Cas9:single-guide RNA (Cas9:sgRNA) ribonucleoprotein (RNP) for genome editing. The structure-activity relationship between the lipidoids and intracellular protein delivery efficiencies was explored and it was demonstrated that the newly developed LNPs are effective for gene recombination in vivo.


Deciphering the Dynamic Complexities of the Liver Microenvironment - Toward a Better Understanding of Immune-Mediated liver Injury Caused by Immune Checkpoint Inhibitors (ILICI).
Immune checkpoint inhibitors (ICIs) represent a promising therapy for many types of cancer. However, only a portion of patients respond to this therapy and some patients develop clinically significant immune-mediated liver injury caused by immune checkpoint inhibitors (ILICI), an immune-related adverse event (irAE) that may require the interruption or termination of treatment and administration of systemic corticosteroids or other immunosuppressive agents. Although the incidence of ILICI is lower with monotherapy, the surge in combining ICIs with chemotherapy, targeted therapy, and combination of different ICIs has led to an increase in the incidence and severity of ILICI - a major challenge for development of effective and safe ICI therapy. In this review, we highlight the importance and contribution of the liver microenvironment to ILICI by focusing on the emerging roles of resident liver cells in modulating immune homeostasis and hepatocyte regeneration, two important decisive factors that dictate the initiation, progression, and recovery from ILICI. Based on the proposed contribution of the liver microenvironment on ICILI, we discuss the clinical characteristics of ILICI in patients with preexisting liver diseases, as well as the challenges of identifying prognostic biomarkers to guide the clinical management of severe ILICI. A better understanding of the liver microenvironment may lead to novel strategies and identification of novel biomarkers for effective management of ILICI.


Multiple component approaches to C-glycosyl beta-amino acids by complementary one-pot Mannich-type and Reformatsky-type reactions.
The development of new methods for the preparation of C-glycosyl beta-amino acid libraries with chemical and stereochemical diversity levels was investigated and the results are described herein. Two complementary one-pot three-component Mannich-type and Reformatsky-type synthetic strategies have been developed for the construction of chiral 3-amino propanoate fragments (eventually bis-substituted at C-2) directly linked to the anomeric carbon of pyranose and furanose residues. Both methods involved as the initial step the coupling of a sugar aldehyde to p-methoxybenzylamine but differed in the nucleophile (a d(2) synthon equivalent) which was successively added: a ketene silyl acetal (Mannich route) or a bromozinc enolate (Reformatsky route). Individual C-glycosyl beta-amino esters were isolated as single 3R diastereoisomers in fair to excellent yield (60-90%) and their structure assigned by NMR spectroscopy (Riguera protocol) supported by X-ray crystallography. A tentative explanation of the observed stereochemical outcome based on transition-state models is provided. A preliminary study on the synthesis of alpha,alpha-difluoro C-glycosyl beta-amino acids via a more traditional Reformatsky route is also reported.


Nodular Cystic Basal Cell Carcinoma of the Trunk: a Diagnostic Dilemma in an Unsuspecting Youth.
Basal cell carcinoma (BCC) commonly affects the elderly and is mostly confined to the head and neck region. Only 10% of all cases occur on the trunk. We presented a case of bullous lesion on the abdomen in a young male, initially diagnosed by clinicians as a vascular nevus. Histopathological examination and immunohistochemistry (IHC) revealed it to be Nodular cystic variant of BCC. This rare variant of BCC morphologically resembles benign skin adnexal tumor of Eccrine syringofibroadenoma. Ber Ep4 positivity on IHC established the correct diagnosis. This case highlights that nodular cystic variant of BCC can be a diagnostic dilemma for clinicians and pathologists.


Machine Learning Prediction and Experimental Validation of Antigenic Drift in H3 Influenza A Viruses in Swine.
The antigenic diversity of influenza A viruses (IAV) circulating in swine challenges the development of effective vaccines, increasing zoonotic threat and pandemic potential. High-throughput sequencing technologies can quantify IAV genetic diversity, but there are no accurate approaches to adequately describe antigenic phenotypes. This study evaluated an ensemble of nonlinear regression models to estimate virus phenotype from genotype. Regression models were trained with a phenotypic data set of pairwise hemagglutination inhibition (HI) assays, using genetic sequence identity and pairwise amino acid mutations as predictor features. The model identified amino acid identity, ranked the relative importance of mutations in the hemagglutinin (HA) protein, and demonstrated good prediction accuracy. Four previously untested IAV strains were selected to experimentally validate model predictions by HI assays. Errors between predicted and measured distances of uncharacterized strains were 0.35, 0.61, 1.69, and 0.13 antigenic units. These empirically trained regression models can be used to estimate antigenic distances between different strains of IAV in swine by using sequence data. By ranking the importance of mutations in the HA, we provide criteria for identifying antigenically advanced IAV strains that may not be controlled by existing vaccines and can inform strain updates to vaccines to better control this pathogen.IMPORTANCE Influenza A viruses (IAV) in swine constitute a major economic burden to an important global agricultural sector, impact food security, and are a public health threat. Despite significant improvement in surveillance for IAV in swine over the past 10 years, sequence data have not been integrated into a systematic vaccine strain selection process for predicting antigenic phenotype and identifying determinants of antigenic drift. To overcome this, we developed nonlinear regression models that predict antigenic phenotype from genetic sequence data by training the model on hemagglutination inhibition assay results. We used these models to predict antigenic phenotype for previously uncharacterized IAV, ranked the importance of genetic features for antigenic phenotype, and experimentally validated our predictions. Our model predicted virus antigenic characteristics from genetic sequence data and provides a rapid and accurate method linking genetic sequence data to antigenic characteristics. This approach also provides support for public health by identifying viruses that are antigenically advanced from strains used as pandemic preparedness candidate vaccine viruses.


Indications and techniques for emergency carotid interventions.
Mutual understanding exists among specialists of neurology, interventional radiology and vascular surgery concerning the need of interventional treatment for patients with symptomatic carotid artery stenosis. However, identification of individuals that will most likely benefit from treatment, timing of the intervention and selection of treatment mode continue to be a matter of intense debate. The aim of this manuscript was to discuss the changing attitude to this high-risk population and present evidence that points towards a resolute, fast-track approach.


Acute Pancreatitis Secondary to Intragastric Balloon: A Case Report and Literature Review.
Intragastric balloon (IGB) is a common minimally invasive procedure used for obesity management and weight reduction. It can be used alone, sequentially, with concomitant therapies, or as a bridge to longer-term weight-loss interventions, such as bariatric surgery. Although the insertion procedure is easy and generally well tolerated by patients, a few complications can occur with varying degrees of severity ranging from mild to severe and life-threatening. Acute pancreatitis is a rare complication of IGB but has been reported in the literature. We present a case in which the patient had a history of IGB insertion complicated by acute pancreatitis. The diagnosis of acute pancreatitis due to the IGB insertion was made after excluding other possible causes of acute pancreatitis. The patient was hospitalized and managed conservatively.


The identity of Cicinnus orthane Blanchard, 1852 (Lepidoptera, Mimallonidae, Cicinninae), type species of Cicinnus Blanchard, 1852.
In order to clarify the identity of the type species of one of the most diverse Mimallonidae genera, Cicinnus Blanchard, 1852, we designate a lectotype for Cicinnus orthane Blanchard, 1852 with photographs of the lectotype (dorsal and ventral) and its genitalia figured for the first time. Cicinnus orthane, described from Chile, is a close morphological match for a southeastern Brazilian species, which we also illustrate for comparative purposes. Because the lectotype of C. orthane is female, we include both sexes of this related Brazilian species in order to facilitate the figuring of male genitalia as well as comparisons of the female genitalia. We are therefore able to establish sound morphological characteristics for Cicinnus sensu stricto, namely in the males the bifid configuration of the gnathos, a complex juxtal arrangement fused to the phallus, and largely membranous valvae; and in the females a wide and narrow lamella antevaginalis, lack of setae covered bulbous masses on either side of the lamella antevaginalis, well-sclerotized and posteriorly protruding tergite VIII, and reduced, stout apophyses anteriores which are about one quarter the length of the apophyses posteriores.


Impaired motor axon regeneration in the C57BL/Ola mouse.
The delayed Wallerian degeneration which occurs in the C57BL/Ola mouse is associated with impaired motor axon regeneration. Following sciatic nerve crush, recovery of the sciatic functional index was delayed and incomplete when compared with recovery in C57BL/6J mice. After facial nerve crush, recovery of whisker movement in Ola mice was also delayed, and there was a prolonged period of partial recovery, not seen in 6J mice. Regeneration rate of the motor axons was measured by the axonal transport technique in sciatic nerve and was approximately 0.7 mm/d for Ola mice, and 4.0 mm/d for 6J mice. Combining these results from our previous work, we conclude that regeneration of both sensory and motor axons is impaired when Wallerian degeneration does not follow its usual time course after injury.


Access to Healthcare and Health Literacy in Croatia: Empirical Investigation.
Health literacy is related to different health-related outcomes. However, the nature of the relationship between health literacy and health outcomes is not well understood. One pathway may lead from health literacy to health outcomes by means of access to healthcare. The goal of the current study is to explore the association between health literacy and the particular measure of access to healthcare-unmet medical need-for the first time in Croatia and, to the best of our knowledge, for the first time in the EU context. We use data obtained from face-to-face interviews in a large nationally representative sample of the Croatian population (n = 1000) to estimate the level of health literacy and self-reported access to care and investigate the association between health literacy and self-perceived barriers to access. Our study showed that limited and problematic health literacy is prevalent and associated with higher rates of unmet medical need. Unmet need is largely caused by long waiting lists. It is therefore essential to design health services fitting the needs of those who have limited and/or problematic health literacy as well as enhance health education with the potential of improving the access to care and health outcomes as well as design policies that reduce waiting times.


Prevalence of and risk factors for hip dysplasia and cranial cruciate ligament deficiency in dogs.
OBJECTIVE-To evaluate prevalence of and risk factors for hip dysplasia (HD) and cranial cruciate ligament deficiency (CCLD) in dogs and determine change in prevalence over time. DESIGN-Cross-sectional study. ANIMALS-1,243,681 Dogs for which information was reported to the Veterinary Medical Database between 1964 and 2003. PROCEDURES-Information on breed, sex, and age was collected, and prevalences and odds ratios were calculated. RESULTS-Castrated male dogs were significantly more likely than other dogs to have HD (odds ratio [OR], 1.21), and castrated male (OR, 1.68) and spayed female (OR, 2.35) dogs were significantly more likely to have CCLD. Dogs up to 4 years old were significantly more likely to have HD (OR for dogs 2 months to 1 year old, 1.22; OR for dogs > 1 to 4 years old, 1.48), whereas dogs > 4 years old were significantly more likely to have CCLD (OR for dogs > 4 to 7 years old, 1.82; OR for dogs > 7 years old, 1.48). In general, large- and giant-breed dogs were more likely than other dogs to have HD, CCLD, or both. Prevalences of HD and CCLD increased significantly over the 4 decades for which data were examined. CONCLUSIONS AND CLINICAL RELEVANCE-Results suggested that sex, age, and breed were risk factors for HD, CCLD, or both in dogs and that prevalences of HD and CCLD have increased over time.


The Internet as a research tool in complementary and alternative medicine: a pilot study.
Information on the use of complementary and alternative medical (CAM) therapies is sometimes difficult to obtain from consumers. The purpose of this study was to evaluate the Internet as a research tool in CAM to obtain information from these consumers about the use of psychotropic herbs and drugs. A survey on psychotropic herb and drug usage was posted on 138 mental-health newsgroups on the Internet. After 38 days, 72 (71%) of the 102 respondents reported usage of psychotropic medication and 68 (67%) reported usage of psychotropic herbs during the preceding 12 months. The Internet is a potential tool to reach consumers of CAM for research purposes. The study design requires modification, and the findings need cautious interpretation for further research using the Internet as a venue.


Association between intraoperative steroid and postoperative mortality in patients undergoing craniotomy for brain tumor.
Despite the widespread use of intraoperative steroids in various neurological surgeries to reduce cerebral edema and other adverse symptoms, there is sparse evidence in the literature for the optimal and safe usage of intraoperative steroid administration in patients undergoing craniotomy for brain tumors. We aimed to investigate the effects of intraoperative steroid administration on postoperative 30-day mortality in patients undergoing craniotomy for brain tumors. Adult patients who underwent craniotomy for brain tumors between January 2011 to January 2020 were included at West China Hospital, Sichuan University in this retrospective cohort study. Stratified analysis based on the type of brain tumor was conducted to explore the potential interaction. This study included 8,663 patients undergoing craniotomy for brain tumors. In patients with benign brain tumors, intraoperative administration of steroids was associated with a higher risk of postoperative 30-day mortality (adjusted OR 1.98, 95% CI 1.09-3.57). However, in patients with malignant brain tumors, no significant association was found between intraoperative steroid administration and postoperative 30-day mortality (adjusted OR 0.86, 95% CI 0.55-1.35). Additionally, administration of intraoperative steroids was not associated with acute kidney injury (adjusted OR 1.11, 95% CI 0.71-1.73), pneumonia (adjusted OR 0.89, 95% CI 0.74-1.07), surgical site infection (adjusted OR 0.78, 95% CI 0.50-1.22) within 30 days, and stress hyperglycemia (adjusted OR 1.05, 95% CI 0.81-1.38) within 24 h after craniotomy for brain tumor. In patients undergoing craniotomy for benign brain tumors, intraoperative steroids were associated with 30-day mortality, but this association was not significant in patients with malignant brain tumors.