At least 10% shorter C-H bonds in cryogenic protein crystal structures than in current AMBER forcefields.
High resolution protein crystal structures resolved with X-ray diffraction data at cryogenic temperature are commonly used as experimental data to refine forcefields and evaluate protein folding simulations. However, it has been unclear hitherto whether the C-H bond lengths in cryogenic protein structures are significantly different from those defined in forcefields to affect protein folding simulations. This article reports the finding that the C-H bonds in high resolution cryogenic protein structures are 10-14% shorter than those defined in current AMBER forcefields, according to 3709 C-H bonds in the cryogenic protein structures with resolutions of 0.62-0.79 Å. Also, 20 all-atom, isothermal-isobaric, 0.5-μs molecular dynamics simulations showed that chignolin folded from a fully-extended backbone formation to the native β-hairpin conformation in the simulations using AMBER forcefield FF12SB at 300 K with an aggregated native state population including standard error of 10 ± 4%. However, the aggregated native state population with standard error reduced to 3 ± 2% in the same simulations except that C-H bonds were shortened by 10-14%. Furthermore, the aggregated native state populations with standard errors increased to 35 ± 3% and 26 ± 3% when using FF12MC, which is based on AMBER forcefield FF99, with and without the shortened C-H bonds, respectively. These results show that the 10-14% bond length differences can significantly affect protein folding simulations and suggest that re-parameterization of C-H bonds according to the cryogenic structures could improve the ability of a forcefield to fold proteins in molecular dynamics simulations.


Endoscopic Treatment of the Zenker Diverticulum With Flexible Endoscopic Myotomy: A Single Tertiary Center Experience.
The Zenker diverticulum (ZD) is the most common type of esophageal diverticula. Management of ZD has different options; however, there is a recent increase in treatment with flexible endoscopic myotomy (FEM). In our study, we aimed to investigate the efficacy and safety of FEM among patients with ZD. The data of patients who underwent FEM for ZD in our clinic between January 2008 and May 2016 were retrospectively analyzed. Myotomy was performed with a needle-knife sphincterotome by using pulse-cut or forced coagulation electrocautery mode. Myotomy was performed on the common wall of the diverticulum up to 0.5 to 1 cm of the distal end. A handmade diverticuloscope, which was modified from an overtube, was used in most of the patients to focus more carefully on the septum. Dysphagia scores of the patients before and after the procedure were compared and complications were evaluated. A total of 17 patients were enrolled in the study. The mean age was 65.3 years, and mean diverticular diameter was 3.2 cm. The most common symptoms were dysphagia and regurgitation. The mean in-hospital stay was 5 days. The dysphagia score was significantly decreased after FEM among the patients (0.17±0.39 vs. 2±0.79; P=0.0001). Complications were observed in 3 patients (17.6%); however, there was no procedure-related mortality. FEM is an efficient and safe procedure as a treatment modality for ZD. However, lack of a standard algorithm is a remarkable disadvantage.


Comparative anatomy and morphology of nectar-producing Melastomataceae.
Most neotropical Melastomataceae have bee-pollinated flowers with poricidal anthers. However, nectar rewards are known to be produced in about 80 species in eight genera from four different tribes. These nectar-producing species are pollinated by both vertebrates and invertebrates. The floral morphology and anatomy of 14 species was studied in six genera of nectar-producing Melastomataceae (Blakea, Brachyotum, Charianthus, Huilaea, Meriania and Miconia). Anatomical methods included scanning electron microscopy, and serial sections of paraffin-embedded flowers. All vertebrate-pollinated melastome flowers have petals that do not open completely at anthesis, thus forming a pseudo-tubular corolla, while closely related species that are bee pollinated have rotate or reflexed corollas. In most species, nectar secretion is related to stomatal or epidermal nectaries and not filament slits as previously reported. Moreover, the nectar is probably supplied by large vascular bundles near the release area. Blakea and Huilaea have nectary stomata located upon the dorsal anther connective appendages. Brachyotum also has nectary stomata on the anther connectives, but these are distributed lengthwise along most of the connective. Meriania may release nectar through the anther connective, but has additional nectary stomata on the inner walls of the hypanthium. Miconia has nectary stomata on the ovary apex. Charianthus nectaries were not found, but there is circumstantial evidence that nectar release occurs through the epidermis at the apex of the ovary and the lower portions of the inner wall of the hypanthium. Nectar release in Melastomataceae is apparently related to nectary stomata and not filament slits. The presence of nectary stomata on stamens and on ovary apices in different lineages suggests that the acquisition of nectaries is a derived condition. Nectary location also supports a derived condition, because location is strongly consistent within each genus, but differs between genera.


Hospice utilization in patients with malignant gliomas.
Despite recommendations from professional organizations supporting early hospice enrollment for patients with cancer, little research exists regarding end-of-life (EOL) practices for patients with malignant glioma (MG). We evaluated rates and correlates of hospice enrollment and hospice length of stay (LOS) among patients with MG. Using the Surveillance, Epidemiology, and End Results (SEER)-Medicare-linked database, we identified adult patients who were diagnosed with MG from January 1, 2002 to December 31, 2011 and who died before December 31, 2012. We extracted sociodemographic and clinical data and used univariate logistic regression analyses to compare characteristics of hospice recipients versus nonrecipients. We performed multivariable logistic regression analyses to examine predictors of hospice enrollment >3 or >7 days prior to death. We identified 12437 eligible patients (46% female), of whom 7849 (63%) were enrolled in hospice before death. On multivariable regression analysis, older age, female sex, higher level of education, white race, and lower median household income predicted hospice enrollment. Of those enrolled in hospice, 6996 (89%) were enrolled for >3 days, and 6047 (77%) were enrolled for >7 days. Older age, female sex, and urban residence were predictors of longer LOS (3- or 7-day minimum) on multivariable analysis. Median LOS on hospice for all enrolled patients was 21 days (interquartile range, 8-45 days). We identified important disparities in hospice utilization among patients with MG, with differences by race, sex, age, level of education, and rural versus urban residence. Further investigation of these barriers to earlier and more widespread hospice utilization is needed.


An overview of atypical enteropathogenic Escherichia coli.
The enteropathogenic Escherichia coli (EPEC) pathotype is currently divided into two groups, typical EPEC (tEPEC) and atypical EPEC (aEPEC). The property that distinguishes these two groups is the presence of the EPEC adherence factor plasmid, which is only found in tEPEC. aEPEC strains are emerging enteropathogens that have been detected worldwide. Herein, we review the serotypes, virulence properties, genetic relationships, epidemiology, reservoir and diagnosis of aEPEC, including those strains not belonging to the classical EPEC serogroups (nonclassical EPEC serogroups). The large variety of serotypes and genetic virulence properties of aEPEC strains from nonclassical EPEC serogroups makes it difficult to determine which strains are truly pathogenic.


Mesenchymal stem cells in the pathogenesis and treatment of bronchopulmonary dysplasia: a clinical review.
Advances in neonatal medicine have led to increased survival of infants born at the limits of viability, resulting in an increased incidence of bronchopulmonary dysplasia (BPD). BPD is a chronic lung disease of premature infants characterized by the arrest of alveolarization, fibroblast activation, and inflammation. BPD leads to significant morbidity and mortality in the neonatal period and is one of the leading causes of chronic lung disease in children. The past decade has brought a surge of trials investigating cellular therapies for the treatment of pulmonary diseases. Mesenchymal stem cells (MSCs) are of particular interest because of their ease of isolation, low immunogenicity, and anti-inflammatory and reparative properties. Clinical trials of MSCs have demonstrated short-term safety and tolerability; however, studies have also shown populations of MSCs with adverse pro-inflammatory and myofibroblastic characteristics. Cell-based therapies may represent the next breakthrough therapy for the treatment of BPD, however, there remain barriers to implementation as well as gaps in knowledge of the role of endogenous MSCs in the pathogenesis of BPD. Concurrent high-quality basic science, translational, and clinical studies investigating the fundamental pathophysiology underlying BPD, therapeutic mechanisms of exogenous MSCs, and logistics of translating cellular therapies will be important areas of future research.


Corticonuclear projections of the cerebellum preserve both anteroposterior and mediolateral pairing patterns.
The aim of the present study was to establish whether a diverging arrangement of the corticonuclear cerebellar projections exists and, if so, what relation it has with the inferior olivary complex. Iontophoretic injections of a 1 : 1 mixture of tetramethylrhodamine dextran amine and biotinylated dextran amine into the cerebellar cortex orthogradely labelled fibre terminals in the cerebellar nuclei and retrogradely labelled cell bodies in the inferior olivary complex. The injections were into A, B, C2, C3, D1 and D2 bands. These injections showed diverging projections to the cerebellar nuclei, with 'primary projections' directed to the nuclear region previously reported to be specifically connected with the injected band and 'secondary projections' directed to other nuclear regions. Secondary projections from the A, C2 and C3 bands diverged to nuclear regions primarily controlled by cortical bands lateral to those injected. Secondary projections from the D1, and D2 bands diverged to nuclear regions primarily controlled by cortical bands medial to those injected. Moreover, injections distributed along the D1 and D2 bands showed similar sets of nuclear targets, while those distributed along the A, C2 and C3 bands showed two sets of nuclear targets in relation to the anteroposterior location of the injected area within these bands. The cortical areas that projected to the same set of nuclear targets were innervated from single olivary regions, while those that projected to different sets of nuclear targets were innervated from different subsets of single regions of the inferior olive. The results suggest that the olivary bands of the cerebellar cortex project to the cerebellar nuclei with a diverging pattern that is organized in both the mediolateral and the anteroposterior axes.


mTOR inhibition as an adjuvant therapy in a metastatic model of HPV+ HNSCC.
Effective treatments for recurrent/metastatic human papillomavirus-positive (HPV+) head and neck squamous cell cancer (HNSCC) are limited. To aid treatment development, we characterized a novel murine model of recurrent/metastatic HPV+ HNSCC. Further analysis of the parental tumor cell line and its four recurrent/metastatic derivatives led to preclinical testing of an effective treatment option for this otherwise fatal disease. Reverse phase protein arrays identified key signaling cascades in the parental and recurrent/metastatic cell lines. While protein expression profiles differed among the recurrent/metastatic cell lines, activated proteins associated with the mTOR signaling cascade were a commonality. Based on these data, mTOR inhibition was evaluated as an adjuvant treatment for recurrent/metastatic disease. mTOR activity and treatment response were assessed in vitro by western blot, Seahorse, proliferation, clonogenic, and migration assays. Standard-of-care cisplatin/radiation therapy (CRT) versus CRT/rapamycin were compared in vivo. Low-dose rapamycin inhibited mTOR signaling, decreasing proliferation (43%) and migration (62%) while it enhanced CRT-induced cytotoxicity (3.3 fold) in clonogenic assays. Furthermore, rapamycin re-sensitized CRT-resistant, metastatic tumors to treatment in vivo, improving long-term cures (0-30% improved to 78-100%, depending on the recurrent/metastatic cell line) and limiting lymph node metastasis (32%) and lung metastatic burden (30 fold). Studies using immune compromised mice suggested rapamycin's effect on metastasis is independent of the adaptive immune response. These data suggest a role of mTOR activation in HPV+ HNSCC recurrent/metastatic disease and that adjuvant mTOR inhibition may enhance treatment of resistant, metastatic cell populations at the primary site and limit distant metastasis.


Study of the B1-B2 transition in colloidal clusters.
The possible mechanisms for the B1 (NaCl-type) to B2 (CsCl-type) transition in crystalline colloidal clusters of equally sized particles are studied by means of two computational techniques: metadynamics and nudged elastic band calculations. The system is modelled by a screened Coulomb potential. Different interaction ranges are considered. The transition from a perfect NaCl cubic cluster to a full CsCl cluster is forced by metadynamics, revealing a transition path with intermediate metastable configurations in which planes are shifted one by one. The presence of metastable configurations in the transition path, corresponding to a certain number of NaCl planes turned into CsCl, has clear analogies with the known Hyde and O'Keeffe mechanism for ionic crystals, with some important differences due to finite-size effects. These comprise the fact that the transition starts by shifting a surface plane by means of a row-by-row mechanism that has no analog in bulk crystals. The energy barriers between the local minima in the transition path are calculated, showing that the barriers strongly depend on the screening length, in such a way that the B1 metastable phase can have very long lifetimes when the interaction is sufficiently long-ranged.


Gene expression ratio test distinguishes normal lung from lung tumors in solid tissue and FNA biopsies.
Lung cancer is the leading cause of cancer-related deaths worldwide. Prognosis and survival are dependent on cell type, early detection, and surgical treatment. Hence, optimal screening strategies and new therapies are urgently required. Although surveillance with low-dose computed tomography can reduce lung cancer mortality by 20%, the number of false-positive detections is significant. Tissue diagnosis aids in the identification of benign nodules, reducing the number of false positive detections. To determine whether molecular testing of fine-needle aspirations (FNAs) can reduce false-positive detections, we developed a gene expression-based test that distinguishes normal from cancerous lung tissues. The test first was applied to published microarray data, showing overall sensitivity and specificity values of 95% (95% CI, 90%-98%) and 100% (95% CI, 40%-100%), respectively. Subsequently, it was validated on 30 solid and ex vivo FNA lung cancer tumor samples and matched normal lung specimens using real-time PCR. The validation test was 93% (95% CI, 78%-99%) sensitive and 100% (95% CI, 88%-100%) specific for the detection of tumor versus normal lung on solid samples, whereas FNA specimens yielded a sensitivity of 91% (95% CI, 72%-99%) and a specificity of 94% (95% CI, 70%-100%). This study supports the hypothesis that the gene-ratio approach reliably distinguishes normal lung from cancerous tissues in FNA samples and can be optimized to diagnose benign nodules.


Pediatric Rheumatology Care and Outcomes Improvement Network's Quality Measure Set to Improve Care of Children With Juvenile Idiopathic Arthritis.
To describe the selection, development, and implementation of quality measures (QMs) for juvenile idiopathic arthritis (JIA) by the Pediatric Rheumatology Care and Outcomes Improvement Network (PR-COIN), a multihospital learning health network using quality improvement methods and leveraging QMs to drive improved outcomes across a JIA population since 2011. An American College of Rheumatology-endorsed multistakeholder process previously selected initial process QMs. Clinicians in PR-COIN and parents of children with JIA collaboratively selected outcome QMs. A committee of rheumatologists and data analysts developed operational definitions. QMs were programmed and validated using patient data. Measures are populated by registry data, and performance is displayed on automated statistical process control charts. PR-COIN centers use rapid-cycle quality improvement approaches to improve performance metrics. The QMs are revised for usefulness, to reflect best practices, and to support network initiatives. The initial QM set included 13 process measures concerning standardized measurement of disease activity, collection of patient-reported outcome assessments, and clinical performance measures. Initial outcome measures were clinical inactive disease, low pain score, and optimal physical functioning. The revised QM set has 20 measures and includes additional measures of disease activity, data quality, and a balancing measure. PR-COIN has developed and tested JIA QMs to assess clinical performance and patient outcomes. The implementation of robust QMs is important to improve quality of care. PR-COIN's set of JIA QMs is the first comprehensive set of QMs used at the point-of-care for a large cohort of JIA patients in a variety of pediatric rheumatology practice settings.


In Vivo Toxicity of Local Anesthetics and Corticosteroids on Chondrocyte and Synoviocyte Viability and Metabolism.
Intra-articular injection of local anesthetic and/or corticosteroid is an adjunct treatment for arthritic and inflammatory orthopedic conditions. Despite potential benefits, there is growing concern that these medications may cause significant morbidity, including potential toxicity to intra-articular chondrocytes and synoviocytes. Twenty dogs underwent intra-articular injection of the shoulder joint using ultrasound guidance, with the following injectates (n = 5 each): negative control (saline), methylprednisolone/1.0% lidocaine, triamcinolone/1.0% lidocaine, and triamcinolone/0.0625% bupivacaine. The dogs were euthanized 24 hours postinjection for reasons unrelated to this study. Synovium/cartilage explants were harvested under sterile conditions and assessed immediately or cultured for 7 days. Synoviocyte and chondrocyte viability was determined on day 1 and day 7 using Calcien AM and Sytox Blue live/dead fluorescent stains, and cell metabolism determined on day 2 using the alamar blue additive test. Results were compared statistically. On day 1 synovium exposed to 1%L/M demonstrated a significant decrease in cell metabolism (P = 0.0107) and subjective synoviocyte viability scores (P = 0.013) compared with the negative control. Cartilage exposed to 1%L/M demonstrated decreased chondrocyte viability and cell metabolism versus all other groups, although not significantly. After 7 days of culture, cartilage viable cell density in the 1%L/M group was significantly (P ≤ 0.001) lower than the negative control. Subjective synoviocyte viability scores was significantly lower in the 1%L/M (P = 0.013), 1%L/T (P ≤ 0.001), and 0.0625%B/T groups (P = 0.006) compared with the negative control. This study suggests potential negative effects of combination local anesthetic/corticosteroid on intra-articular cell viability and cell metabolism. Further study is needed before determining definitive clinical recommendations.


Anterior cruciate ligament insufficiency in children.
Anterior cruciate ligament (ACL) insufficiency is unusual in children younger than 14 years of age. Nontraumatic ACL laxity is found in two distinct groups of children: (1) those with generalized nonpathologic joint laxity, and (2) those with congenital absence or attenuation of the ligament, usually associated with other congenital anomalies in the same limb. ACL insufficiency secondary to trauma most often is associated with avulsion of the intercondylar eminence of the tibia. Excellent results have been obtained following anatomic reduction by either closed or open methods. In children, insubstance damage to the ACL, alone or associated with other ligamentous injuries, is distinctly unusual. In three patients younger than 14 years of age with midsubstance tears of the ACL, no other clinically evident ligamentous injury was noted. All three were treated by primary surgical repair without intra- or extra-articular augmentation procedures. Despite only mild subjective complaints at follow-up examinations, all three patients had at least moderate degrees of clinical ACL laxity, suggesting that midsubstance ACL injuries in children have no better healing potential than in adults.


Recursive optimal pruning with applications to tree structured vector quantizers.
A pruning algorithm of P.A. Chou et al. (1989) for designing optimal tree structures identifies only those codebooks which lie on the convex hull of the original codebook's operational distortion rate function. The authors introduce a modified version of the original algorithm, which identifies a large number of codebooks having minimum average distortion, under the constraint that, in each step, only modes having no descendents are removed from the tree. All codebooks generated by the original algorithm are also generated by this algorithm. The new algorithm generates a much larger number of codebooks in the middle- and low-rate regions. The additional codebooks permit operation near the codebook's operational distortion rate function without time sharing by choosing from the increased number of available bit rates. Despite the statistical mismatch which occurs when coding data outside the training sequence, these pruned codebooks retain their performance advantage over full search vector quantizers (VQs) for a large range of rates.


Psychotropic drug prescription patterns among patients with bipolar I disorder.
Combination treatment, rather than monotherapy, is prevalent in the treatment of subjects with bipolar disorder, probably due to the complex and phasic nature of the illness. In general, prescription patterns may be influenced by the demographic characteristics of patients as well. We evaluated prescription patterns and the influence of demographic variables on these patterns in a voluntary registry of subjects with bipolar disorder. A subset of data from a larger voluntary registry was extracted for demographic variables and psychotropic medication use that had been reported in the month prior to registration by ambulatory, non-hospitalized subjects with bipolar I disorder in 1995/96 (n = 457). Among the thymoleptic agents, lithium was prescribed in over 50% of subjects, valproate in approximately 40%, and carbamazepine in 11% of subjects. Eighteen percent of subjects had no prescription for thymoleptic agents. Nearly one-third of all subjects were receiving antipsychotic agents, of whom two-thirds were receiving the traditional neuroleptic agents. More than half of all subjects were receiving concomitant antidepressants, of whom nearly 50% received the SSRI antidepressants and nearly 25% received buproprion. Approximately 40% of subjects received benzodiazepines. Only 18% of subjects received monotherapy, and nearly 50% received three or more psychotropic agents. In general, no associations were noted between demographic parameters including age, gender, marital or educational status, and psychotropic prescriptions. Consistent with the anecdotal reports, these data confirm that combination treatment is far more common than monotherapy. Demography appears to have a minimal impact on cross-sectional prescription patterns in subjects with bipolar disorder. Given that combination treatments are the rule rather than the exception, we should strive to achieve rational, yet pragmatic, treatment guidelines and algorithms to minimize the risks while maximizing the benefits of these combination treatments for patients with bipolar disorder.


Meta-analysis of epilepsy prevalence in West Africa and its relationship with onchocerciasis endemicity and control.
A high prevalence and incidence of epilepsy has been reported in onchocerciasis-endemic regions in Central and East Africa. There is compelling epidemiological evidence suggesting that this high burden is caused by onchocerciasis-associated epilepsy (OAE). We hypothesized that OAE had also occured in West African onchocerciasis foci. We searched PubMed, the African Journals Online platform and grey literature for population-based epilepsy studies in West African countries. Epilepsy and onchocerciasis prevalence data were extracted. The pre-control onchocerciasis endemicity in the study sites was estimated from historical data of onchocerciasis control programmes. The prevalence of epilepsy in different sites was analysed, taking into account onchocerciasis endemicity and the duration of control. The pooled prevalence of epilepsy in the West African study sites was 13.14 per 1000 (95% confidence interval 11.28-15.00). Higher pre-control endemicity and a shorter duration of onchocerciasis control were both associated with increased epilepsy prevalence (p<0.001). Two studies in Ivory Coast that provided detailed descriptions of persons with epilepsy in onchocerciasis-endemic settings revealed that most of them had features of OAE (73.7% and 83.3%, respectively). Our findings suggest that before and during the early years of implementing onchocerciasis control in West Africa, high onchocerciasis endemicity resulted in a high prevalence of OAE and that subsequent control efforts significantly reduced the prevalence of OAE.


Oligosaccharides in molecular recognition.
The development, organization and growth of complex organisms as well as their interactions with the environment involve an intricate array of molecular recognition events. There is an increased awareness of the involvement of oligosaccharides in many of these processes. In this article, studies of oligosaccharide antigenicity, and the way these have been interpreted with respect to oligosaccharide function will be discussed. In addition, examples of oligosaccharides as receptor, first, as receptors and determinants of susceptibility to an exogenous infective agent and secondly, as recognition structures possibly involved in endogenous interactions, will be described. This will be followed by a discussion of the recent hypothesis in which oligosaccharides are envisaged as recognition structures and integral components of cell growth-regulating networks. Finally, an outline of new strategies for decoding the information content in glycoprotein oligosaccharides will be given.


Involvement of dachshund and Distal-less in distal pattern formation of the cricket leg during regeneration.
Cricket nymphs have the remarkable ability to regenerate a functional leg following amputation, indicating that the regenerating blastemal cells contain information for leg morphology. However, the molecular mechanisms that underlie regeneration of leg patterns remain poorly understood. Here, we analyzed phenotypes of the tibia and tarsus (three tarsomeres) obtained by knockdown with regeneration-dependent RNA interference (rdRNAi) against Gryllus dachshund (Gb'dac) and Distal-less (Gb'Dll). We found that depletion of Gb'Dll mRNA results in loss of the tarsal segments, while rdRNAi against Gb'dac shortens the tibia at the two most distal tarsomeres. These results indicate that Gb'Dll expression is indispensable for formation of the tarsus, while Gb'dac expression is necessary for elongation of the tibia and formation of the most proximal tarsomere. These findings demonstrate that mutual transcriptional regulation between the two is indispensable for formation of the tarsomeres, whereas Gb'dac is involved in determination of tibial size through interaction with Gb'ds/Gb'ft.


A systematic review of measurements of physical function in critically ill adults.
Patients who recover from critical illness may be left with significant limitations to their physical function that can have important consequences for their quality of life. Measures of physical function may be useful end points to consider in studies conducted in critically ill patients and are particularly attractive in studies investigating early mobilisation and rehabilitation. To describe measurements of physical function used in studies investigating early mobilisation and rehabilitation in critically ill adults. A systematic search of Ovid MEDLINE, Embase, CINAHL, Cochrane Library and PEDro was undertaken to identify studies investigating early mobilisation and rehabilitation in critically ill adults. Two researchers independently extracted data from identified studies that described measurements of physical function and that evaluated the available evidence for the measurement properties and risk of bias associated with the identified end points. We identified 11 studies of early mobilisation and rehabilitation in critically ill patients, involving 19 distinct measures of physical function. The ability to perform activities such as sitting and standing and the maximum distance ambulated were the most commonly used end points. Only one end point in the included studies, the Functional Status Score for the ICU (FSS-ICU), was designed for use in the ICU setting. Of the end points used, only the Short Form 36 (SF-36), the Medical Research Council (MRC) scale score and handheld dynamometers have proven interrater reliability and population validity in the ICU setting. A wide range of end points have been used to evaluate physical function in critically ill patients. However, further studies are needed to establish the measurement properties of the most commonly used end points in order to recommend their use in clinical trials.


Evaluation of benign vs malignant hepatic lesions with positron emission tomography.
In most malignant cells, the relatively low level of glucose-6-phosphatase leads to accumulation and trapping of [18F]fluorodeoxyglucose (FDG) intracellularly, allowing the visualization of increased uptake compared with normal cells. To assess the value of FDG positron emission tomography (PET) to differentiate benign from malignant hepatic lesions and to determine in which types of hepatic tumors PET can help evaluate stage, monitor response to therapy, and detect recurrence. Prospective blinded-comparison clinical cohort study. Tertiary care university hospital and clinic. One hundred ten consecutive referred patients with hepatic lesions 1 cm or larger on screening computed tomographic (CT) images who were seen for evaluation and potential resection underwent PET imaging. There were 60 men and 50 women with a mean (+/-SD) age of 59 +/- 14 years. Follow-up was 100%. A PET scan using static imaging was performed on all patients. The PET scan imaging and biopsy, surgery, or both were performed, providing pathological samples within 2 months of PET imaging. All PET images were correlated with CT scan to localize the lesion. However, PET investigators were unaware of any previous interpretation of the CT scan. Visual interpretation, lesion-to-normal liver background (L/B) ratio of radioactivity, and standard uptake value (SUV) were correlated with pathological diagnosis. All (100%) liver metastases from adenocarcinoma and sarcoma primaries in 66 patients and all cholangiocarcinomas in 8 patients had increased uptake values, L/B ratios greater than 2, and an SUV greater than 3.5. Hepatocellular carcinoma had increased FDG uptake in 16 of 23 patients and poor uptake in 7 patients. All benign hepatic lesions (n = 23), including adenoma and fibronodular hyperplasia, had poor uptake, an L/B ratio of less than 2, and an SUV less than 3.5, except for 1 of 3 abscesses that had definite uptake. The PET technique using FDG static imaging was useful to differentiate malignant from benign lesions in the liver. Limitations include false-positive results in a minority of abscesses and false-negative results in a minority of hepatocellular carcinoma. The PET technique was useful in tumor staging and detection of recurrence, as well as monitoring response to therapy for all adenocarcinomas and sarcomas and most hepatocellular carcinomas. Therefore, pretherapy PET imaging is recommended to help assess new hepatic lesions.


Predicting leaf area index from scaling principles: corroboration and consequences.
Leaf area index (LAI) is a key biophysical variable in most process-based forest-ecosystem models. However, most such models require LAI as an input, typically obtained from empirical observations. We tested whether scaling principles based on trade-offs between single leaf and canopy properties could be effectively used to model LAI, thereby obviating the need for empirical observations. To do so, we used the process-oriented model, PnET, configured to estimate LAI from these same scaling principles. We derived biologically based LAI predictions (LAIPnET) for the Harvard Forest (Massachusetts, USA) eddy covariance tower site, a predominately mixed deciduous hardwood forest, using PnET, and compared these with a locally observed phenology record and with LAI estimates from both local (ground-based) photosynthetically active radiation transmittance (LAITRANS) and normalized difference vegetation index satellite data (LAINDVI). We generated the LAIPnET trajectory by running the PnET model with meteorological observations from the flux tower as model drivers. We derived LAITRANS from measurements of above- and below-canopy photosynthetically active radiation at the flux tower, and LAINDVI from observations from the Advanced Very High Resolution Radiometer (AVHRR) satellite-borne sensor of surface greenness for the 1 km2 cell containing the flux tower. Over a 5-year period, LAIPnET and LAITRANS values were comparable intra- and interannually, with maximum values differing by less than 0.1 to 0.2 LAI units (m2 m(-2)). Values of LAINDVI were similar to LAIPnET and LAITRANS in midsummer, but higher LAI values were predicted in the early and late portions of the growing season. In addition, we used the three alternative LAI trajectories in a modified version of the PnET model and compared the resulting outputs of gross primary production (GPP) with GPP estimates from the flux tower for 5 continuous years. The LAIPnET and LAITRANS inputs resulted in a difference of less than 3% in mean annual GPP from 1995 to 1999, and these were within 7 and 9%, respectively, of the annual eddy flux-based estimates over the same time period. The results indicate that biologically based LAI scaling approaches can closely track temporal changes in a deciduous forest and have potential for spatial and temporal scaling of LAI.


Oseltamivir ring prophylaxis for containment of 2009 H1N1 influenza outbreaks.
From June 22 through June 25, 2009, four outbreaks of infection with the pandemic influenza A (H1N1) virus occurred in Singapore military camps. We report the efficacy of ring chemoprophylaxis (geographically targeted containment by means of prophylaxis) with oseltamivir to control outbreaks of 2009 H1N1 influenza in semiclosed environments. All personnel with suspected infection were tested and clinically isolated if infection was confirmed. In addition, we administered postexposure ring chemoprophylaxis with oseltamivir and segregated the affected military units to contain the spread of the virus. All personnel were screened three times weekly both for virologic infection, by means of nasopharyngeal swabs and reverse-transcriptase-polymerase-chain-reaction assay with sequencing, and for clinical symptoms, by means of questionnaires. A total of 1175 personnel were at risk across the four sites, with 1100 receiving oseltamivir prophylaxis. A total of 75 personnel (6.4%) were infected before the intervention, and 7 (0.6%) after the intervention. There was a significant reduction in the overall reproductive number (the number of new cases attributable to the index case), from 1.91 (95% credible interval, 1.50 to 2.36) before the intervention to 0.11 (95% credible interval, 0.05 to 0.20) after the intervention. Three of the four outbreaks showed a significant reduction in the rate of infection after the intervention. Molecular analysis revealed that all four outbreaks were derived from the New York lineage of the 2009 H1N1 virus and that cases within each outbreak were due to transmission rather than unrelated episodes of infection. Of the 816 personnel treated with oseltamivir who were surveyed, 63 (7.7%) reported mild, nonrespiratory side effects of the drug, with no severe adverse events. Oseltamivir ring chemoprophylaxis, together with prompt identification and isolation of infected personnel, was effective in reducing the impact of outbreaks of 2009 H1N1 influenza in semiclosed settings.


Immunohistochemical and immunoelectron microscopic study of cytochrome P-450 of human fetal livers (P-450HFLa): implications for an onco-feto-placental enzyme.
Cytochrome P-450 of human fetal livers (P-450HFLa) was demonstrated by the avidin-biotin immunoperoxidase technique in tissue samples as follows: human fetal organs, adult livers, human and cynomolgus placenta, and gynecologic organs which were obtained from 40 patients with gynecologic malignancies and 32 patients with benign diseases. P-450HFLa was clearly localized in the cytoplasm and membranes of the hepatocytes, and the fact was confirmed by an immunoelectron microscopic examination. In addition, a semiquantitative assay of staining intensity demonstrated that this enzyme tended to decrease with advancing age. These findings suggest that hepatic P-450HFLa synthesis is inversely proportional to age, and that this enzyme is one of the differentiation antigens. P-450HFLa was also detected immunohistochemically in other fetal organs. The present study thus confirms that P-450HFLa is not specific to the liver and is ubiquitous even in the fetus. Marked positive staining for P-450HFLa was demonstrated in villous syncytiotrophoblasts. In contrast, no positive staining was found in the cynomolgus-monkey placenta, unlike the case for many other placental antigens. These findings lead to the tentative conclusion that P-450HFLa is a feto-placental enzyme peculiar to humans. P-450HFLa was demonstrated to occur very frequently in gynecologic malignancies. The mean positivity rate for all gynecologic malignancies was 85%, while the rate was below 25% for benign gynecologic diseases, indicating that P-450HFLa is one of the onco-feto-placental enzymes. The present study thus suggests that this enzyme could be a promising new tumor marker for gynecologic malignancies.


Submandibular ductal fistula: an obstacle to sialendoscopy.
Submandibular fistula is a rare condition that presents diagnostic and treatment challenges. Accurate diagnosis may be achieved by means of thorough physical examination, proper imaging, and usually surgical exploration. We describe the clinical significance of aberrant submandibular ductal fistula or ostium and discuss our dilemmas during sialendoscopy in 2 patients with repeated submandibular glandular swelling. Two patients had submandibular ductal abnormalities. The first patient was found to have the ostium of the submandibular duct in an unusual anatomical location at the posterior floor of the mouth, which was identified during surgical exploration of the floor of the mouth. The second patient was found to have a submandibular ductal fistula into the floor of the mouth proximal to a calculus in the main duct (which was severely atrophic and could not be cannulated). The precise etiology and pathogenesis of sialo-oral fistula formation are currently unknown but could be extrapolated from previously described syndromes involving ductal obstruction and inciting damage in other regions of the body. Consideration of submandibular fistula in the differential diagnosis may spare the patient morbidity of redundant invasive procedures.


1H NMR-based metabonomic investigation of tributyl phosphate exposure in rats.
Tributyl phosphate (TBP) is a toxic organophosphorous compound widely used in many industrial applications, including significant usage in nuclear processing. The industrial application of this chemical is responsible for occupational exposure and environmental pollution. In this study, (1)H NMR-based metabonomics has been applied to investigate the metabolic response to TBP exposure. Male Sprague-Dawley rats were given a TBP-dose of 15 mg/kg body weight, followed by 24h urine collection, as was previously demonstrated for finding most of the intermediates of TBP. High-resolution (1)H NMR spectroscopy of urine samples in conjunction with statistical pattern recognition and compound identification allowed for the metabolic changes associated with TBP treatment to be identified. Discerning NMR spectral regions corresponding to three TBP metabolites, dibutyl phosphate (DBP), N-acetyl-(S-3-hydroxybutyl)-L-cysteine and N-acetyl-(S-3-oxobutyl)-L-cysteine, were identified in TBP-treated rats. In addition, the (1)H NMR spectra revealed TBP-induced variations of endogenous urinary metabolites including benzoate, urea, and trigonelline along with metabolites involved in the Krebs cycle including citrate, cis-aconitate, trans-aconitate, 2-oxoglutarate, succinate, and fumarate. These findings indicate that TBP induces a disturbance to the Krebs cycle energy metabolism and provides a biomarker signature of TBP exposure. We show that three metabolites of TBP, dibutylphosphate, N-acetyl-(S-3-hydroxybutyl)-L-cysteine and N-acetyl-(S-3-oxobutyl)-L-cysteine, which are not present in the control groups, are the most important factors in separating the TBP and control groups (p<0.0023), while the endogenous compounds 2-oxoglutarate, benzoate, fumarate, trigonelline, and cis-aconetate were also important (p<0.01).


The Actin Cytoskeleton in SMA and ALS: How Does It Contribute to Motoneuron Degeneration?
Amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA) are neurodegenerative diseases with overlapping clinical phenotypes based on impaired motoneuron function. However, the pathomechanisms of both diseases are largely unknown, and it is still unclear whether they converge on the molecular level. SMA is a monogenic disease caused by low levels of functional Survival of Motoneuron (SMN) protein, whereas ALS involves multiple genes as well as environmental factors. Recent evidence argues for involvement of actin regulation as a causative and dysregulated process in both diseases. ALS-causing mutations in the actin-binding protein profilin-1 as well as the ability of the SMN protein to directly bind to profilins argue in favor of a common molecular mechanism involving the actin cytoskeleton. Profilins are major regulat ors of actin-dynamics being involved in multiple neuronal motility and transport processes as well as modulation of synaptic functions that are impaired in models of both motoneuron diseases. In this article, we review the current literature in SMA and ALS research with a focus on the actin cytoskeleton. We propose a common molecular mechanism that explains the degeneration of motoneurons for SMA and some cases of ALS.


The bidirectional relation of emotion perception and social judgments: the effect of witness' emotion expression on perceptions of moral behaviour and vice versa.
The present research tested the notion that emotion expression and context perception are bidirectionally related. Specifically, in two studies focusing on moral violations (N = 288) and positive moral deviations (N = 245) respectively, we presented participants with short vignettes describing behaviours that were either (im)moral, (in)polite or unusual together with a picture of the emotional reaction of a person who supposedly had been a witness to the event. Participants rated both the emotional reactions observed and their own moral appraisal of the situation described. In both studies, we found that situational context influenced how emotional reactions to this context were rated and in turn, the emotional expression shown in reaction to a situation influenced the appraisal of the situation. That is, neither the moral events nor the emotion expressions were judged in an absolute fashion. Rather, the perception of one also depended on the other.


The tarsal tunnel syndrome in children.
There are few reports on the tarsal tunnel syndrome in children. This paper concerns 10 such children. In adults the syndrome is equally distributed among the sexes but all these children were girls. Trauma preceded the symptoms in only two cases. The symptoms differed in some aspects from those usually seen in adults. Six of the children walked with the affected foot in supination. Three of the six, and one other, used crutches at intervals. All were operated on and at follow-up nine were symptom-free and the tenth had improved.


MiR-134/487b/655 cluster regulates TGF-β-induced epithelial-mesenchymal transition and drug resistance to gefitinib by targeting MAGI2 in lung adenocarcinoma cells.
Epithelial-mesenchymal transition (EMT) has recently been recognized as a key element of cell invasion, migration, metastasis, and drug resistance in several types of cancer, including non-small cell lung cancer (NSCLC). Our aim was to clarify microRNA (miRNA)-related mechanisms underlying EMT followed by acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) in NSCLC. miRNA expression profiles were examined before and after transforming growth factor β1 (TGF-β1) exposure in four human adenocarcinoma cell lines with or without EMT. Correlation between expressions of EMT-related miRNAs and resistance to EGFR-TKI gefitinib was evaluated. miRNA array and real-time quantitative reverse transcription PCR (qRT-PCR) revealed that TGF-β1 significantly induced overexpression of miR-134, miR-487b, and miR-655, which belong to the same cluster located on chromosome 14q32, in lung adenocarcinoma cells with EMT. MAGI2 (membrane-associated guanylate kinase, WW, and PDZ domain-containing protein 2), a predicted target of these miRNAs and a scaffold protein required for PTEN, was diminished in A549 cells with EMT after the TGF-β1 stimulation. Overexpression of miR-134 and miR-487b promoted the EMT phenomenon and affected the drug resistance to gefitinib, whereas knockdown of these miRNAs inhibited the EMT process and reversed TGF-β1-induced resistance to gefitinib. Our study demonstrated that the miR-134/487b/655 cluster contributed to the TGF-β1-induced EMT phenomenon and affected the resistance to gefitinib by directly targeting MAGI2, in which suppression subsequently caused loss of PTEN stability in lung cancer cells. The miR-134/miR-487b/miR-655 cluster may be a new therapeutic target in patients with advanced lung adenocarcinoma, depending on the EMT phenomenon.


CHARACTERIZING CALCIUM INFLUX VIA VOLTAGE- AND LIGAND-GATED CALCIUM CHANNELS IN EMBRYONIC ALLIGATOR NEURONS IN CULTURE.
Vertebrate brains share many features in common. Early in development, both the hindbrain and diencephalon are built similarly. Only later in time do differences in morphology occur. Factors that could potentially influence such changes include certain physiological properties of neurons. As an initial step to investigate this problem, embryonic Alligator brain neurons were cultured and calcium responses were characterized. The present report is the first to document culture of Alligator brain neurons in artificial cerebrospinal fluid (ACSF) as well as in standard mammalian tissue culture medium supplemented with growth factors. Alligator brain neuron cultures were viable for at least 1 week with unipolar neurites emerging by 24 hours. Employing Fura-2 AM, robust depolarization-induced calcium influx, was observed in these neurons. Using selective blockers of the voltage-gated calcium channels, the contributions of N-, P/Q-, R-, T-, and L-type channels in these neurons were assessed and their presence documented. Lastly, Alligator brain neurons were challenged with an excitotoxic stimulus (glutamate + glycine) where delayed calcium deregulation could be prevented by a classical NMDA receptor antagonist.


Raman spectrum of supercritical C(18)O2 and re-evaluation of the Fermi resonance.
We report the first Raman spectra of fully (18)O-labeled supercritical CO(2) (scCO(2)) and various isotopic mixtures. The experimental results, coupled with ab initio molecular dynamics calculations, demonstrate that the frequencies assigned to the Fermi dyad of the CO(2) molecule transpose upon isotopic labeling of both oxygen atoms. Although the transposition of the Fermi dyad of CO(2) gas due to isotopic substitution has been discussed before, this is the first confirmation of the effect in the Raman spectrum of the supercritical fluid and provides necessary groundwork for future Raman spectroscopy studies of reactions in this important medium. More importantly, the work yields a quantitative assessment of the mixing of states upon labeling that provides the needed clarification concerning the pedigree of the assignments for the dyad of CO(2) under supercritical conditions.


Nucleic Acids Delivery Into the Cells Using Pro-Apoptotic Protein Lactaptin.
Cell penetrating peptides (CPP) are promising agents for transporting diverse cargo into the cells. The amino acid sequence and the mechanism of lactaptin entry into the cells allow it to be included into CPP group. Lactaptin, the fragment of human milk kappa-casein, and recombinant lactaptin (RL2) were initially discovered as molecules that induced apoptosis of cultured cancer cells and did not affect non-malignant cells. Here, we analyzed the recombinant lactaptin potency to form complexes with nucleic acids and to act as a gene delivery system. To study RL2-dependent delivery, three type of nucleic acid were used as a models: plasmid DNA (pDNA), siRNA, and non-coding RNA which allow to detect intracellular localization through their functional activity. We have demonstrated that RL2 formed positively charged noncovalent 110-nm-sized complexes with enhanced green fluorescent protein (EGFP)-expressing plasmid DNA. Ca2+ ions stabilized these complexes, whereas polyanion heparin displaced DNA from the complexes. The functional activity of delivered nucleic acids were assessed by fluorescent microscopy using A549 lung adenocarcinoma cells and A431 epidermoid carcinoma cells. We observed that RL2:pDNA complexes provided EGFP expression in the treated cells and that strongly confirmed the entering pDNA into the cells. The efficiency of cell transformation by these complexes increased when RL2:pDNA ratio increased. Pre-treatment of the cells with anti-RL2 antibodies partly inhibited the entry of pDNA into the cells. RL2-mediated delivery of siRNA against EGFP was analyzed when A549 cells were co-transfected with EGFP-pDNA and RL2:siRNA complexes. siRNA against EGFP efficiently inhibited the expression of EGFP being delivered as RL2:siRNA complexes. We have previously demonstrated that non-coding U25 small nucleolar RNA (snoRNA) can decrease cell viability. Cancer cell transfection with RL2-snoRNA U25 complexes lead to a substantial decrease of cell viability, confirming the efficiency of snoRNA U25 delivery. Collectively, these findings indicate that recombinant lactaptin is able to deliver noncovalently associated nucleic acids into cancer cells in vitro.


Radon daughter levels in some public and private buildings in India.
Radon daughter concentrations have been measured in some public and private buildings of Aligarh city. Approximately 320 CR-39 detectors were mounted in 30 sample sites. It was found that the Rn daughter concentrations vary from 3.5 mWL to 8.1 mWL with a geometric mean of 5.6 +/- 1.3 mWL. The average annual effective dose equivalent due to Rn daughters is found to be 1.46 mSv using an equilibrium factor of 0.45 and an occupancy factor of 0.8. About 33% of buildings were estimated to have a Rn concentration below 40 Bq m-3 and 79% below 60 Bq m-3. The measured levels do not require any intervention as recommended by the U.S. Environmental Protection Agency.


Complement anaphylatoxins C3a and C5a induce a failing regenerative program in cardiac resident cells. Evidence of a role for cardiac resident stem cells other than cardiomyocyte renewal.
Cardiac healing, which follows myocardial infarction, is a complex process guided by intricate interactions among different components. Some resident cell populations with a potential role in cardiac healing have already been described in cardiac tissues. These non-cardiomyocyte cell subsets, globally described as cardiac pluripotent/progenitor cells (CPCs), are able to differentiate into all three major cardiac cell lineages (endothelial, smooth muscle and cardiomyocyte cells) in experimental settings. Nevertheless, physiological cardiac healing results in a fibrous scar, which remains to be fully modelled experimentally. Since a role for complement anaphylatoxins (C3a and C5a) has been described in several regeneration/repair processes, we examined the effects that C3a and C5a exert on a defined population of CPCs. We found that C3a and C5a are able to enhance CPC migration and proliferation. In vitro studies showed that this effect is linked to activation of telomerase mRNA and partial preservation of telomere length, in an NFκB-dependent manner. In addition, anaphylatoxin signalling modulates the CPC phenotype, increasing myofibroblast differentiation and reducing endothelial and cardiac gene expression. These findings may denote that C3a and C5a are able to maintain/increase the cardiac stem cell pool within the heart, whilst simultaneously facilitating and modulating resident cell differentiation. We found that this modulation was directed towards scar forming cells, which increased fibroblast/myofibroblast generation and suggests that both these anaphylatoxins could play a relevant role in the damage-coupled activation of resident cells, and regulation of the cardiac healing process after injury.


Volatile Flavor Profile and Sensory Properties of Vegetable Soybean.
The volatile flavor profiles and sensory properties of different vegetable soybean varieties popularized and cultivated in China for 20, 10, and 2 years (TW292, X3, and SX6, respectively) were investigated. Nutrient composition analysis revealed that TW292 had a high soluble protein and soluble sugar content but low fat content. The total free amino acid content (15.43 mg/g) and umami free amino acid content (6.08 mg/g) of SX6 were significantly higher (p < 0.05) than those of the other varieties. An electronic tongue effectively differentiated between the umami and sweetness characteristics of the vegetable soybeans. Differences in sensory evaluation results were mainly reflected in texture and taste. A total of 41 volatile compounds were identified through HS-SPME-GC-MS, and the main flavor compounds were 1-octen-3-ol, hexanal, (Z)-2-heptenal, 2-octene, nonanal, (Z)-2-decenal, and 3,5-octadien-2-one. However, the volatile composition of different vegetable soybean varieties exhibited large variability in type and relative contents. Considerable differences in nutritional, organoleptic, and aroma characteristics were found among different varieties. The results of this study will provide a good basis for the assessment and application of the major vegetable soybean varieties grown in China.


A Critical Care Clinician Survey Comparing Attitudes and Perceived Barriers to Low Tidal Volume Ventilation with Actual Practice.
Low-Vt ventilation lowers mortality in patients with acute respiratory distress syndrome (ARDS) but is underused. Little is known about clinician attitudes toward and perceived barriers to low-Vt ventilation use and their association with actual low-Vt ventilation use. The objectives of this study were to assess clinicians' attitudes toward and perceived barriers to low-Vt ventilation (Vt <6.5 ml/kg predicted body weight) in patients with ARDS, to identify differences in attitudes and perceived barriers among clinician types, and to compare attitudes toward and perceived barriers to actual low-Vt ventilation use in patients with ARDS. We conducted a survey of critical care physicians, nurses, and respiratory therapists at four non-ARDS Network hospitals in the Chicago region. We compared survey responses with performance in a cohort of 362 patients with ARDS. Survey responses included clinician attitudes toward and perceived barriers to low-Vt ventilation use. We also measured low-Vt ventilation initiation by these clinicians in 347 patients with ARDS initiated after ARDS onset as well as correlation with clinician attitudes and perceived barriers. Of 674 clinicians surveyed, 467 (69.3%) responded. Clinicians had positive attitudes toward and perceived few process barriers to ARDS diagnosis or initiation of low-Vt ventilation. Physicians had more positive attitudes and perceived fewer barriers than nurses or respiratory therapists. However, use of low-Vt ventilation by all three clinician groups was low. For example, whereas physicians believed that 92.5% of their patients with ARDS warranted treatment with low-Vt ventilation, they initiated low-Vt ventilation for a median (interquartile range) of 7.4% (0 to 14.3%) of their eligible patients with ARDS. Clinician attitudes and perceived barriers were not correlated with low-Vt ventilation initiation. Clinicians had positive attitudes toward low-Vt ventilation and perceived few barriers to using it, but attitudes and perceived process barriers were not correlated with actual low-Vt ventilation use, which was low. Implementation strategies should be focused on examining other issues, such as ARDS recognition and process solutions, to improve low-Vt ventilation use.


XAV939: from a small inhibitor to a potent drug bioconjugate when delivered by gold nanoparticles.
Nanoparticles as potential drug delivery vectors are drawing more attention every day. Here, we used gold nanopspheres (AuNSs) to selectively target the Wnt signaling pathway in human oral squamous cell carcinoma (HSC-3) cells. In a previously conducted study, XAV939, a small inhibiter, was found to strongly regulate the Wnt pathway by inhibiting the tankyrase enzyme and subsequent stabilization of cytoplasmic axin levels. In the present study, conjugating XAV939 molecules to AuNSs is found to enhance its potency by at least 100 times over its free form in killing HSC-3 cancer cells. Additionally, XAV 939 uptake studies have demonstrated an enhanced XAV939 bioconjugate delivery to the targeted cells compared to the passive cellular diffusion of the free drug at the same concentration. Furthermore, our study revealed that drug delivery and cytotoxicity are directly related to the size of the functionalized nanoparticles.


Abnormal patterns of theta frequency oscillations during the temporal evolution of freezing of gait in Parkinson's disease.
We sought to characterize the electrophysiological signature of Freezing of gait in Parkinson's disease. We examined 24 patients with idiopathic Parkinson's disease and significant freezing of gait as they performed a series of timed up-and-go tasks in their 'off' state while electroencephalographic data was collected from four scalp leads. Fast Fourier Transformation was utilized to explore the power spectral density between periods of normal walking and periods of freezing, as well as during the transition between the two states. In addition, Cross Spectrum and Cross Frequency analyses were used to explore the role of impaired temporal and spatial connectivity. When compared to walking, episodes of freezing were associated with a significant increase in theta band power within the central and frontal leads. The transition from normal walking to freezing of gait was also associated with increased theta frequency coupling between the central and frontal leads, along with an increase in cross-frequency coupling in the central lead. Episodes of freezing of gait in Parkinson's disease are associated with abnormal oscillatory activity in the brain. These results provide novel insights into the pattern of spatiotemporal dynamics underlying freezing of gait and may provide a potential means for therapeutic prediction and alleviation of freezing episodes in susceptible patients.


Depolarized light scattering and dielectric response of a peptide dissolved in water.
The density and orientational relaxation of bulk water can be separately studied by depolarized light scattering (DLS) and dielectric spectroscopy (DS), respectively. Here, we ask the question of what are the leading collective modes responsible for polarization anisotropy relaxation (DLS) and dipole moment relaxation (DS) of solutions involving mostly hydrophobic solute-water interfaces. We study, by atomistic molecular dynamics simulations, the dynamics and structure of hydration water interfacing N-Acetyl-leucine-methylamide dipeptide. The DLS response of the solution is consistent with three relaxation processes: bulk water, rotations of single solutes, and collective dipole-induced-dipole polarizability of the solutes, with the time-scale of 130-200 ps. No separate DLS response of the hydration shell has been identified by our simulations. Density fluctuations of the hydration layer, which largely contribute to the response, do not produce a dynamical process distinct from bulk water. We find that the structural perturbation of the orientational distribution of hydration waters by the dipeptide solute is quite significant and propagates ∼3-5 hydration layers into the bulk. This perturbation is still below that produced by hydrated globular proteins. Despite this structural perturbation, there is little change in the orientational dynamics of the hydration layers, compared to the bulk, as probed by both single-particle orientational dynamics and collective dynamics of the dipole moment of the shells. There is a clear distinction between the perturbation of the interfacial structure by the solute-solvent interaction potential and the perturbation of the interfacial dynamics by the corresponding forces.


A brief history of enviroethics and its challenges.
Environmental ethics has emerged during the early 1970s, when environmentalists started urging philosophers to consider the philosophical aspects of environmental problems. Environmental ethics considers the ethical relationships between humanity and non-human world. The Union of Concerned Scientists, a group of over two thousands scientists, has concluded that climatic change is beyond dispute, and already changing our environment. Environmental instability portend ill for public health and well-being. This paper attempts to apply ethical theories to support environmental concerns and provides moral grounds to preserve the earth's environment. This article documents consensus among environmental philosophers as given by synthesis data provided via survey among articles, websites, and books by the keywords: environment, ethics, health and crises. The field has come to exert significant influence over a large number of human science disciplines in relation to environmental sustainability and human wellbeing. Environmental ethics focuses on the possibility of the identification of human ego with nature, means the larger ecological self deserves respect, too. Environmental ethics expands the boundaries of ethics to include the nature and considers its sustainability to ensure human wellbeing. This study emphasizes mainly on a brief history of environmental ethics and its protection against damage. Environmental changes and extreme weather events in plus to species distinction and a growth of diseases are impossible to hide and ought to be impossible to ignore. The health decline associated with various forms of these changes is continuing. It raises crucial issues about environmental justice.


Hematopoietic stem cells produce intermediate lineage adipocyte progenitors that simultaneously express both myeloid and mesenchymal lineage markers in adipose tissue.
Some adipocytes are produced from bone marrow hematopoietic stem cells. In vitro studies previously indicated that these bone marrow-derived adipocytes (BMDAs) were generated from adipose tissue macrophage (ATM) that lose their hematopoietic markers and acquire mesenchymal markers prior to terminal adipogenic differentiation. Here we interrogated whether this hematopoietic-to-mesenchymal transition drives BMDA production In vitro. We generated transgenic mice in which the lysozyme gene promoter (LysM) indelibly labeled ATM with green fluorescent protein (GFP). We discovered that adipose stroma contained a population of LysM-positive myeloid cells that simultaneously expressed hematopoietic/myeloid markers (CD45 and CD11b), and mesenchymal markers (CD29, PDGFRa and Sca-1) typically found on conventional adipocyte progenitors. These cells were capable of adipogenic differentiation In vitro and In vitro, while other stromal populations deficient in PDGFRa and Sca-1 were non-adipogenic. BMDAs and conventional adipocytes expressed common fat cell markers but exhibited little or no expression of hematopoietic and mesenchymal progenitor cell markers. The data indicate that BMDAs are produced from ATM simultaneously expressing hematopoietic and mesenchymal markers rather than via a stepwise hematopoietic-to-mesenchymal transition. Because BMDA production is stimulated by high fat feeding, their production from hematopoietic progenitors may maintain adipocyte production when conventional adipocyte precursors are diminished.


The Impact of State Firearm Laws on Homicide Rates among Black and White Populations in the United States, 1991-2016.
This study aimed to investigate the potential differential effects of state-level firearm laws on black and white populations. Using a panel design, authors examined the relationship between state firearm laws and homicide victimization rates among white people and black people in 39 states during the period between 1991 and 2016. Authors modeled homicide rates using linear regression with year and state fixed effects and controlled for a range of time-varying, state-level factors. Results showed that universal background check laws and permit requirement laws were associated with lower homicide rates among both white and black populations, and "shall issue" laws were associated with higher homicide rates among both white and black populations. Laws that prohibit firearm possession among people convicted of a violent misdemeanor or require relinquishment of firearms by people with a domestic violence restraining order were associated with lower black homicide rates, but not with white homicide rates. Author identification of heterogeneity in the associations between state firearm laws and homicide rates among different racial groups has implications for reducing racial health disparities.


5-Aminolevulinic acid dehydratase. The role of sulphydryl groups in 5-aminolevulinic acid dehydratase from bovine liver.
The thiophilic reagent 5,5'-dithiobis(2-nitrobenzoic acid) Nbs2) reacts with four sulphydryl groups in native 5-aminolevulinic acid dehydratase from bovine liver (groups I, II, III and IV). All four of these groups exhibit various degrees of half-site reactivity. Groups I and II are highly reactive and their rates of reaction with Nbs2 have been investigated using stopped-flow analysis. The reaction of these groups with Nbs2 results in the formation of an intramolecular disulphide bond which may be reduced with dithioerythritol to regenerate the free sulphydryl groups. Groups I and II appear to be at, or near, the catalytic site whereas group III is involved in the maintenance of conformation in the native enzyme.


Characteristics and referral of emergency department patients with elevated blood pressure.
To assess the prevalence, characteristics, and referral to health care of emergency department (ED) patients with elevated blood pressure (BP) and to compare those without a history of hypertension (HTN) with suboptimally controlled, known-hypertensive patients. A prospective, observational study was conducted in non-critically ill adults with a triage systolic blood pressure (sBP) of > or =140 or with a diastolic blood pressure (dBP) of > or =90 mmHg. Two additional measurements were obtained, and a standardized questionnaire recorded demographics, medical history, and ED staff referrals. Of 1,719 nonpregnant adults, 991 agreed to participate, of whom 455 had elevated triage BP values. Using the mean of three measurements, 305 (67%) had an sBP of > or =140 or a dBP of > or =90 mm Hg (mean sBP = 157 and mean dBP = 89 mm Hg). Subjects with no prior history of HTN (n = 105) were predominantly male, younger, and more likely to smoke and consume alcohol than were known-hypertensive patients (n = 200). Three quarters of all subjects had access to primary health care; however, time elapsed since the last BP assessment was greater (155 vs. 53 d, p = 0.03) in subjects with no previous history. These subjects were also less likely to be informed of their elevated BP (33% vs. 50%, p = 0.02) or to be instructed by ED staff to obtain a repeat measurement (13% vs. 31%, p = 0.001). Of ED patients with elevated BP, one third had no prior history, and the remainder were suboptimally controlled hypertensive patients. Both groups were inadequately informed of their elevated BPs, and the group with no prior history of HTN, the population likely to obtain the greatest health benefit, had the lowest referral rate.


Risk factors for osteoarthritis: understanding joint vulnerability.
Joint damage occurs when structures protecting the joint fail. Because osteoarthritis consists of end-stage joint damage, I propose that risk factors for disease can be best understood as either impairments of joint protectors, increasing joint vulnerability, or as factors that excessively load the joint, leading to injury. I review current knowledge using this paradigm, separating factors into those increasing joint vulnerability (malalignment, muscle weakness, genetic and ethnic predispositions, aging) and those that cause excessive loading (obesity; certain physical activities). Osteoarthritis and cartilage loss can occur without pain. This review focuses separately on factors associated with pain in those with osteoarthritis. To identify sources of pain in osteoarthritis, recent studies have compared magnetic resonance imaging findings of people with osteoarthritis with and without pain, focusing on structures known to have nociceptive innervations. Those with pain are more likely to have effusions, bone marrow lesions, synovial hypertrophy, and tendinitis and bursitis around the joint. This review creates a new paradigm for understanding risk factors for osteoarthritis, using joint vulnerability and loading as a framework and focuses separately on the emerging investigative area of sources of pain.


Spontaneous Activity Drives Local Synaptic Plasticity In Vivo.
Spontaneous activity fine-tunes neuronal connections in the developing brain. To explore the underlying synaptic plasticity mechanisms, we monitored naturally occurring changes in spontaneous activity at individual synapses with whole-cell patch-clamp recordings and simultaneous calcium imaging in the mouse visual cortex in vivo. Analyzing activity changes across large populations of synapses revealed a simple and efficient local plasticity rule: synapses that exhibit low synchronicity with nearby neighbors (<12 μm) become depressed in their transmission frequency. Asynchronous electrical stimulation of individual synapses in hippocampal slices showed that this is due to a decrease in synaptic transmission efficiency. Accordingly, experimentally increasing local synchronicity, by stimulating synapses in response to spontaneous activity at neighboring synapses, stabilized synaptic transmission. Finally, blockade of the high-affinity proBDNF receptor p75(NTR) prevented the depression of asynchronously stimulated synapses. Thus, spontaneous activity drives local synaptic plasticity at individual synapses in an "out-of-sync, lose-your-link" fashion through proBDNF/p75(NTR) signaling to refine neuronal connectivity. VIDEO ABSTRACT.


A life-course perspective on physical functioning in women.
To validate Kalache & Kickbusch's model: namely, that functional capacity peaks in early adulthood, then declines at a rate dependent on fitness level until a "disability threshold" is reached. Data came from the Australian Longitudinal Study on Women's Health, which followed three cohorts from 1996 to 2011: a young, a mid-aged and an older cohort (born in 1973-78, 1946-51 and 1921-26, respectively). The Short Form (36) Health Survey was used to measure physical functioning (score 1-100). The disability threshold was the mean physical functioning score in older women requiring assistance with daily activities (62.8). The relationship between age and physical functioning was modelled using spline regression for the entire sample, and by baseline physical functioning quintile and physical activity level. PHYSICAL DECLINE QUICKENED WITH AGE: 0.05 annual units (95% confidence interval, CI: -0.13 to 0.22) at ages 18-23 years (i.e. no decline); -2.43 (95% CI: -2.64 to -2.23) at ages 82-90 years. Decline was faster in quintiles with lower baseline physical functioning in the younger and mid-age cohorts and in quintiles with higher baseline physical functioning in the older cohort. The disability threshold was reached at a mean age of 79 years, but the range was 45-88 years, depending on baseline physical functioning and physical activity. Age and physical decline are not linearly related, as traditionally believed; decline accelerates with age. However, baseline physical functioning, but not physical activity, influences the rate of decline.


Chronic pancreatitis associated with the p.G208A variant of PRSS1 gene in a European patient.
The major etiologic factor of chronic pancreatitis in adults is excessive alcohol consumption, whereas among children structural anomalies, systemic and metabolic disorders, and genetic factors are prevalent. Mutations in the cationic trypsinogen gene (PRSS1) cause hereditary pancreatitis, while mutations in serine protease inhibitor Kazal type 1 (SPINK1), cystic fibrosis transmembrane conductance regulator (CFTR) and chymotrypsin C (CTRC) genes have been shown to associate with chronic pancreatitis as independent risk factors. We present a case of 13-year-old boy with idiopathic chronic pancreatitis. Given the unexplained attacks of pancreatitis since early childhood and despite the negative family history, molecular-genetic analysis of four pancreatitis susceptibility genes (PRSS1, SPINK1, CTRC and CFTR) was performed. The boy was found to carry the c.623G>C (p.G208A) mutation of the PRSS1 gene and the c.180C>T (p.G60G) mutation of the CTRC gene, both in heterozygous state. These mutations are considered as contributing risk factors for chronic pancreatitis. In children with idiopathic chronic pancreatitis genetic causes should be considered, even in absence of positive family history. To the best of our knowledge, this is the first description of a European patient with chronic pancreatitis associated with the p.G208A mutation of PRSS1 gene. This mutation was previously reported only in Asian subjects and is thought to be a unique genetic cause of pancreatitis in Asia.


Hypoxia-induced endocytosis of Na,K-ATPase in alveolar epithelial cells is mediated by mitochondrial reactive oxygen species and PKC-zeta.
During ascent to high altitude and pulmonary edema, the alveolar epithelial cells (AEC) are exposed to hypoxic conditions. Hypoxia inhibits alveolar fluid reabsorption and decreases Na,K-ATPase activity in AEC. We report here that exposure of AEC to hypoxia induced a time-dependent decrease of Na,K-ATPase activity and a parallel decrease in the number of Na,K-ATPase alpha(1) subunits at the basolateral membrane (BLM), without changing its total cell protein abundance. These effects were reversible upon reoxygenation and specific, because the plasma membrane protein GLUT1 did not decrease in response to hypoxia. Hypoxia caused an increase in mitochondrial reactive oxygen species (ROS) levels that was inhibited by antioxidants. Antioxidants prevented the hypoxia-mediated decrease in Na,K-ATPase activity and protein abundance at the BLM. Hypoxia-treated AEC deficient in mitochondrial DNA (rho(0) cells) did not have increased levels of ROS, nor was the Na,K-ATPase activity inhibited. Na,K-ATPase alpha(1) subunit was phosphorylated by PKC in hypoxia-treated AEC. In AEC treated with a PKC-zeta antagonist peptide or with the Na,K-ATPase alpha(1) subunit lacking the PKC phosphorylation site (Ser-18), hypoxia failed to decrease Na,K-ATPase abundance and function. Accordingly, we provide evidence that hypoxia decreases Na,K-ATPase activity in AEC by triggering its endocytosis through mitochondrial ROS and PKC-zeta-mediated phosphorylation of the Na,K-ATPase alpha(1) subunit.


Association between markers of systemic inflammation, oxidative stress, lipid profiles, and insulin resistance in pregnant women.
Increased levels of pro-inflammatory factors, markers of oxidative stress and lipid profiles are known to be associated with several complications. The aim of this study was to determine the association of markers of systemic inflammation, oxidative stress and lipid profiles with insulin resistance in pregnant women in Kashan, Iran. In a cross-sectional study, serum high sensitivity C-reactive protein (hs-CRP), tumor necrosis factor-alpha (TNF-α), fasting plasma glucose (FPG), serum insulin, 8-oxo-7, 8-dihydroguanine (8-oxo-G), total cholesterol, triglyceride, High density lipoprotein-cholesterol (HDL-cholesterol), and plasma total antioxidant capacity (TAC) were measured among 89 primigravida singleton pregnant women aged 18-30 years at 24-28 weeks of gestation. Pearson's correlation and multiple linear regressions were used to assess their relationships with homeostatic model assessment of insulin resistance (HOMA-IR). We found that among biochemical indicators of pregnant women, serum hs-CRP and total cholesterol levels were positively correlated with HOMA-IR (β = 0.05, P = 0.006 for hs-CRP and β = 0.006, P = 0.006 for total cholesterol). These associations remained significant even after mutual effect of other biochemical indicators were controlled (β = 0.04, P = 0.01 for hs-CRP and β = 0.007, P = 0.02 for total cholesterol). Further adjustment for body mass index made the association of hs-CRP and HOMA-IR disappeared; however, the relationship for total cholesterol remained statistically significant. Our findings showed that serum total cholesterol is independently correlated with HOMA-IR score. Further studies are needed to confirm our findings.


Inducible plasmid-determined resistance to arsenate, arsenite, and antimony (III) in escherichia coli and Staphylococcus aureus.
Plasmids in both Escherichia coli and Staphylococcus aureus contain an "operon" that confers resistance to arsenate, arsenite, and antimony(III) salts. The systems were always inducible. All three salts, arsenate, arsenite, and antimony(III), were inducers. Mutants and a cloned deoxyribonucleic acid fragment from plasmid pI258 in S. aureus have lost arsenate resistance but retained resistances to arsenite and antimony, demonstrating that separate genes are involved. Arsenate-resistant arsenite-sensitive S. aureus plasmid mutants were also isolated. In E. coli, plasmid-determined arsenate resistance and reduced uptake were additive to that found with chromosomal arsenate resistance mutants. Arsenate resistance was due to reduced uptake of arsenate by the induced plasmid-containing cells. Under conditions of high arsenate, when some uptake could be demonstrated with the induced resistant cells, the arsenate was rapidly lost by the cells in the absence of extracellular phosphate. Sensitive cells retained arsenate under these conditions. When phosphate was added, phosphate-arsenate exchange occurred. High phosphate in the growth medium protected cells from arsenate, but not from arsenite or antimony(III) toxicity. We do not know the mechanisms of arsenite or antimony resistance. However, arsenite was not oxidized to less toxic arsenate. Since cell-free medium "conditioned" by prior growth to induced resistant cells with toxic levels of arsenite or antimony(III) retained the ability to inhibit the growth of sensitive cells, the mechanism of arsenite and antimony resistance does not involve conversion of AsO2- or SbO+ to less toxic forms or binding by soluble thiols excreted by resistant cells.


Systems Biology Analysis for Ewing Sarcoma.
Ewing sarcoma (EwS) is a highly aggressive pediatric bone cancer that is defined by a somatic fusion between the EWSR1 gene and an ETS family member, most frequently the FLI1 gene, leading to expression of a chimeric transcription factor EWSR1-FLI1. Otherwise, EwS is one of the most genetically stable cancers. The situation when the major cancer driver is well known looks like a unique opportunity for applying the systems biology approach in order to understand the EwS mechanisms as well as to uncover some general mechanistic principles of carcinogenesis. A number of studies have been performed revealing the direct and indirect effects of EWSR1-FLI1 on multiple aspects of cellular life. Nevertheless, the emerging picture of the oncogene action appears to be highly complex and systemic, with multiple reciprocal influences between the immediate consequences of the driver mutation and intracellular and intercellular molecular mechanisms, including regulation of transcription, epigenome, and tumoral microenvironment. In this chapter, we present an overview of existing molecular profiling resources available for EwS tumors and cell lines and provide an online comprehensive catalogue of publicly available omics and other datasets. We further highlight the systems biology studies of EwS, involving mathematical modeling of networks and integration of molecular data. We conclude that despite the seeming simplicity, a lot has yet to be understood on the systems-wide mechanisms connecting the driver mutation and the major cellular phenotypes of this pediatric cancer. Overall, this chapter can serve as a guide for a systems biology researcher to start working on EwS.


Lewy body-related alpha-synucleinopathy in the aged human brain.
To clarify the significance of Lewy body (LB)-related alpha-synucleinopathy in aging and various neurodegenerative disorders, its incidence and topographic pattern were examined in 260 brains of elderly patients, including 116 autopsy-proven cases of Alzheimer disease (AD), 71 cases of clinically and autopsy-proven Parkinson disease (PD), 38 of dementia with Lewy bodies (DLB), 8 patients with progressive supranuclear palsy (PSP), one with senile tremor, and 26 age-matched controls without neuropsychiatric disorders. Using immunohistochemistry, alpha-synuclein (AS) positive lesions were assessed semiquantitatively. For technical reasons, the olfactory system was not systematically studied. All PD-brains showed AS-positive lesions in medullary, pontine and mesencephalic nuclei, with involvement of the nucleus basalis (90.1%), limbic cortex (58.9%), cingulate cortex (46%), amygdala, CA 2/3 hippocampal region (36.2%), neocortex (28.8%), and striatum (11%). 88% of clinical PD cases corresponded to LB pathology stages 4-6, 12% to stage 3 according to Braak et al. (2003). 84% of DLB brains were PD stage 5 or 6 and 17% stage 4, without significant differences between DLB with and without neuritic AD pathology, suggesting morphologic similarities betwee these disorders. 6/8 PSP and senile tremor cases, 49.1% of AD and 69% of aged controls were negative. AS-positive lesions in AD showed decreasing incidence from midbrain (24-28%), limbic cortex and amygdala (17-18%), nucleus basalis and medullary nuclei (13-17%), cingulate cortex (12%), CA 2/3 region (8%) to neocortex (2%), without gender differences or relationship to the severity of AD pathology (mean Braak stage 5.1). AD cases with AS positive lesions, particularly those with AS pathology in the amygdala, were older at death than negative ones (86.6 vs 83.3 yrs), but this difference was not statistically significant. 15 AD cases (seven of them with mild PD symptoms) and 3 aged controls without parkinsonian signs but LB pathology stages 3 (n=5) and 4 (n=13) were considered "incidental LB disease". 16 AD brains without parkinsonian symptoms had AS positive lesions in various areas without medullary involvement, suggesting deviation from the proposed stereotypic expansion pattern. Located AS-pathology in the midbrain and limbic cortex was seen in 31% of asymptomatic aged controls. These data 1. largely confirm Braak's staging of LB-pathology in PD; 2. suggest morphologic and pathogenic relations between PD (brainstem type) and DLB with and without coexistent AD pathology; 3. the occurrence of LB-related alpha-synucleinopathy in about 50% of AD brains and about 30% of aged controls. However, the basic mechanisms of LB-related AS-pathology and their pathogenic and clinical relevance in aged brain and neurodegenerative disorders await further elucidation.


Half-hedgehog spin textures in sub-100 nm soft magnetic nanodots.
Topologically non-trivial structures such as magnetic skyrmions are nanometric spin textures of outstanding potential for spintronic applications due to their unique features. It is well known that Néel skyrmions of definite chirality are stabilized by the Dzyaloshinskii-Moriya exchange interaction (DMI) in bulk non-centrosymmetric materials or ultrathin films with strong spin-orbit coupling at the interface. In this work, we show that soft magnetic (permalloy) hemispherical nanodots are able to host three-dimensional chiral structures (half-hedgehog spin textures) with non-zero tropological charge. They are observed at room temperature, in absence of DMI interaction and they can be further stabilized by the magnetic field arising from the Magnetic Force Microscopy probe. Micromagnetic simulations corroborate the experimental data. Our work implies the existence of a new degree of freedom to create and manipulate complex 3D spin-textures in soft magnetic nanodots and opens up future possibilities to explore their magnetization dynamics.


The diagnostic utility of the keratin profiles of hepatocellular carcinoma and cholangiocarcinoma.
A total of 32 hepatocellular carcinomas (HCC), 10 cholangiocarcinomas (CC), one combined HCC-CC, and 10 adenocarcinomas metastatic to the liver were studied immunohistochemically using AE1 and Cam 5.2, monoclonal antikeratin antibodies with different specificities. AE1 recognizes keratins with molecular weights of 56.5, 50/50', 48, and 40 kd (keratin nos. 10, 14, 15, 16, and 19, according to Moll's catalog), and labels many epithelia, including bile duct epithelium, but not hepatocytes. Both biliary epithelium and hepatocytes are stained by Cam 5.2, which reacts with keratins of molecular weights 50, 43, and 39 kd (corresponding to keratin nos. 8, 18, and 19). Tissues were formalin fixed, paraffin embedded, and a three-stage immunoperoxidase technique was employed. Of 32 pure HCCs, 29 were unreactive with AE1 yet were positive with Cam 5.2. The intensity and extent of immunostaining with Cam 5.2 did not correlate with tumor grade. In contrast to the HCCs, all 10 CCs and the 10 hepatic metastases were strongly positive with both AE1 and Cam 5.2. The combined HCC-CC was also labeled by both antibodies. We conclude that most HCCs express an immunohistochemical keratin profile identical to that of nonneoplastic hepatocytes, which differs from the keratin patterns of bile ducts, CCs, and metastatic adenocarcinomas from a variety of primary sites. These differences in immunoreactivity with antikeratin antibodies may prove useful in diagnostic surgical pathology.


The nitrogen requirement of the weanling kitten.
The nitrogen requirement of the weanling kitten was determined in a series of three experiments. In each experiment, diets were formulated to provide the growing kitten with the essential amino acids at or above the level of requirement. Expt 1 utilized a 4 x 4 balanced Latin square design with two groups of kittens (four male and four female). The crystalline L-amino acid diets were presented at four levels of dietary crude protein (N X 6.25) of 140, 160, 180 and 200 g/kg diet. The design for Expts 2 and 3 was a 6 X 6 balanced Latin square. For each of these experiments, groups of six male and six female kittens were assigned to diets. The six levels of dietary crude protein were 120, 140, 160, 180, 200 and 220 g/kg diet; dietary N was supplied by crystalline L-amino acids for Expt 2 and casein plus a supplementary amino acid mix for Expt 3. Food intake, weight gain and N retention were determined in each experiment. A sigmoidal model y = P1 + P2/[1 + e(P3 + P4 X chi] was fitted to the response of weight gain and N retention to dietary N. The calculated requirement (95% of the upper asymptote, P1 + P2) for these experiments varied from 170 to 230 g protein/kg diet with the majority of these values falling between 180 and 200 g protein/kg. On the basis of these three experiments, the kitten's requirement for dietary crude protein is between 180 and 200 g/kg diet (28.8-32.0 gN/kg) for purified diets which provide a calculated 21 MJ metabolizable energy/kg diet.


Concept Maps as a Tool for Teaching Organic Chemical Reactions.
The purpose of the research was to establish the impact of the application of a concept map in chemistry lessons on the effective solving of tasks with organic reactions content. In the first phase of the research, a concept map was produced representing the reactions of hydrocarbons, organic halogenated compounds and organic oxygen compounds, and in the second phase the produced concept map was introduced in lessons. Its impact was tested on a sample consisting of 186 students (average age of 17.8 years), who were divided into a control group (88 students) and an experimental group (98 students). Prior to the experiment, the two groups were equalised in terms of their level of development of formal logical thinking and their average grade in chemistry. A knowledge test, consisting of five problem tasks comprising multiple parts, was used as a quantitative instrument for measuring the impact of the applied concept map. The content of the knowledge test was selected on the basis of the chemistry lesson plan (reactions of organic oxygen compounds) for general upper secondary schools (in Slovenian: gimnazije). An analysis of the task solving showed statistically significant differences in the responses of the experimental group members and the control group members (experimental group M = 15.9; SD = 6.33; control group M = 13.6; SD = 7.93; p = 0.03). The produced concept map contributed to the more effective interrelation of concepts and, consequently, to more effective problem task solving.


Endocannabinoid Gene × Gene Interaction Association to Alcohol Use Disorder in Two Adolescent Cohorts.
Genetic markers of the endocannabinoid system have been linked to a variety of addiction-related behaviors that extend beyond cannabis use. In the current study we investigate the relationship between endocannabinoid (eCB) genetic markers and alcohol use disorder (AUD) in European adolescents (14-18 years old) followed in the IMAGEN study (n = 2,051) and explore replication in a cohort of North American adolescents from Canadian Saguenay Youth Study (SYS) (n = 772). Case-control status is represented by a score of more than 7 on the Alcohol Use Disorder Identification Test (AUDIT). First a set-based test method was used to examine if a relationship between the eCB system and AUDIT case/control status exists at the gene level. Using only SNPs that are both independent and significantly associated to case-control status, we perform Fisher's exact test to determine SNP level odds ratios in relation to case-control status and then perform logistic regressions as post-hoc analysis, while considering various covariates. Generalized multifactor dimensionality reduction (GMDR) was used to analyze the most robust SNP×SNP interaction of the five eCB genes with positive AUDIT screen. While no gene-sets were significantly associated to AUDIT scores after correction for multiple tests, in the case/control analysis, 7 SNPs were significantly associated with AUDIT scores of > 7 (p < 0.05; OR<1). Two SNPs remain significant after correction by false discovery rate (FDR): rs9343525 in CNR1 (pcorrected =0.042, OR = 0.73) and rs507961 in MGLL (pcorrected = 0.043, OR = 0.78). Logistic regression showed that both rs9353525 (CNR1) and rs507961 (MGLL) remained significantly associated with positive AUDIT screens (p < 0.01; OR < 1) after correction for multiple covariables and interaction of covariable × SNP. This result was not replicated in the SYS cohort. The GMDR model revealed a significant three-SNP interaction (p = 0.006) involving rs484061 (MGLL), rs4963307 (DAGLA), and rs7766029 (CNR1) predicted case-control status, after correcting for multiple covariables in the IMAGEN sample. A binomial logistic regression of the combination of these three SNPs by phenotype in the SYS cohort showed a result in the same direction as seen in the IMAGEN cohort (BETA = 0.501, p = 0.06). While preliminary, the present study suggests that the eCB system may play a role in the development of AUD in adolescents.


Serum paraoxonase-1 activity is more closely related to HDL particle concentration and large HDL particles than to HDL cholesterol in Type 2 diabetic and non-diabetic subjects.
We determined relationships of the anti-oxidative enzyme, paraoxonase-1 (PON-1), with high density lipoprotein (HDL) subfractions, and tested whether these relationships are stronger than those with HDL cholesterol and apolipoprotein A-I (apoA-I) in subjects with and without type 2 diabetes mellitus (T2DM). Serum PON-1 (arylesterase activity) and HDL subfractions (nuclear magnetic resonance spectroscopy) were determined in 67 T2DM patients and in 56 non-diabetic subjects. PON-1 activity, HDL cholesterol and apoA-I were decreased in T2DM (all p<0.05). The HDL particle concentration was unaltered, but large HDL particles, medium HDL particles and HDL particle size were decreased, whereas small HDL particles were increased in T2DM (all p<0.05). PON-1 was more closely related to HDL cholesterol than to apoA-I (p=0.001). In turn, the positive relationship of PON-1 with the HDL particle concentration and with large HDL particles was stronger than that with HDL cholesterol (both p<0.01). The inverse relationship of PON-1 with T2DM was only modestly attenuated by HDL cholesterol or HDL particle characteristics. PON-1 activity is more closely related to the HDL particle concentration or large HDL particles than to HDL cholesterol. Impaired PON-1 activity in T2DM is not to a considerable extent explained by altered HDL subfraction levels.


Donor-derived cell-free DNA as a biomarker for rejection after kidney transplantation: a systematic review and meta-analysis.
A systematic review and meta-analysis were performed to investigate the value of donor-derived cell-free DNA (dd-cfDNA) as a noninvasive biomarker in diagnosing kidney allograft rejection. We searched PubMed, Web of Science and the Cochrane Library for original research papers published between January 1994 and May 2020 on dd-cfDNA fractions in blood of kidney allograft recipients. A single-group meta-analysis was performed by computing pooled estimates for dd-cfDNA fractions using the weighted median of medians or quantile estimation (QE) approach. Weighted median differences in medians (WMDMs) and median differences based on the QE method were used for pairwise comparisons. Despite heterogeneity among the selected studies, the meta-analysis revealed significantly higher median dd-cfDNA fractions in patients with antibody-mediated rejection (ABMR) than patients without rejection or patients with stable graft function. When comparing patients with T cell-mediated rejection (TCMR) and patients with ABMR, our two statistical approaches revealed conflicting results. Patients with TCMR did not have different median dd-cfDNA fractions than patients without rejection or patients with stable graft function. dd-cfDNA may be a useful marker for ABMR, but probably not for TCMR.


Resuscitation with amiodarone increases survival after hemorrhage and ventricular fibrillation in pigs.
Supplemental digital content is available in the text. The aim of this experimental study was to compare survival and hemodynamic effects of a low-dose amiodarone and vasopressin compared with vasopressin in hypovolemic cardiac arrest model in piglets. Eighteen anesthetized male piglets (with a weight of 25.3 [1.8] kg) were bled approximately 30% of the total blood volume via the femoral artery to a mean arterial blood pressure of 35 mm Hg in a 15-minute period. Afterward, the piglets were subjected to 4 minutes of untreated ventricular fibrillation followed by 11 minutes of open-chest cardiopulmonary resuscitation. At 5 minutes, circulatory arrest amiodarone 1 mg/kg was intravenously administered in the amiodarone group (n = 9), while the control group received the same amount of saline (n = 9). At the same time, all piglets received vasopressin 0.4 U/kg intravenously administered and hypertonic-hyperoncotic solution 3-mL/kg infusion for 20 minutes. Internal defibrillation was attempted from 7 minutes of cardiac arrest to achieve restoration of spontaneous circulation. The experiment was terminated 3 hours after resuscitation. Three-hour survival was greater in the amiodarone group (p = 0.02). After the successful resuscitation, the amiodarone group piglets had significantly lower heart rate as well as greater systolic, diastolic, and mean arterial pressure. Troponin I plasma concentrations were lower and urine output was greater in the amiodarone group. Combined resuscitation with amiodarone and vasopressin after hemorrhagic circulatory arrest resulted in greater 3-hour survival, better preserved hemodynamic parameters, and smaller myocardial injury compared with resuscitation with vasopressin only.