Lung texture in serial thoracic computed tomography scans: correlation of radiomics-based features with radiation therapy dose and radiation pneumonitis development.
To assess the relationship between radiation dose and change in a set of mathematical intensity- and texture-based features and to determine the ability of texture analysis to identify patients who develop radiation pneumonitis (RP). A total of 106 patients who received radiation therapy (RT) for esophageal cancer were retrospectively identified under institutional review board approval. For each patient, diagnostic computed tomography (CT) scans were acquired before (0-168 days) and after (5-120 days) RT, and a treatment planning CT scan with an associated dose map was obtained. 32- × 32-pixel regions of interest (ROIs) were randomly identified in the lungs of each pre-RT scan. ROIs were subsequently mapped to the post-RT scan and the planning scan dose map by using deformable image registration. The changes in 20 feature values (ΔFV) between pre- and post-RT scan ROIs were calculated. Regression modeling and analysis of variance were used to test the relationships between ΔFV, mean ROI dose, and development of grade ≥2 RP. Area under the receiver operating characteristic curve (AUC) was calculated to determine each feature's ability to distinguish between patients with and those without RP. A classifier was constructed to determine whether 2- or 3-feature combinations could improve RP distinction. For all 20 features, a significant ΔFV was observed with increasing radiation dose. Twelve features changed significantly for patients with RP. Individual texture features could discriminate between patients with and those without RP with moderate performance (AUCs from 0.49 to 0.78). Using multiple features in a classifier, AUC increased significantly (0.59-0.84). A relationship between dose and change in a set of image-based features was observed. For 12 features, ΔFV was significantly related to RP development. This study demonstrated the ability of radiomics to provide a quantitative, individualized measurement of patient lung tissue reaction to RT and assess RP development.


Noncancerous Genitourinary Conditions as a Public Health Priority: Conceptualizing the Hidden Burden.
To provide a conceptual framework to guide investigations into burdens of noncancerous genitourinary conditions (NCGUCs), which are extensive and poorly understood. The National Institute of Diabetes and Digestive and Kidney Diseases convened a workshop of diverse, interdisciplinary researchers and health professionals to identify known and hidden burdens of NCGUCs that must be measured to estimate the comprehensive burden. Following the meeting, a subgroup of attendees (authors of this article) continued to meet to conceptualize burden. The Hidden Burden of Noncancerous Genitourinary Conditions Framework includes impacts across multiple levels of well-being and social ecology, including individual (ie, biologic factors, lived experience, behaviors), interpersonal (eg, romantic partners, family members), organizational/institutional (eg, schools, workplaces), community (eg, public restroom infrastructure), societal (eg, health care and insurance systems, national workforce/economic output), and ecosystem (eg, landfill waste) effects. The framework acknowledges that NCGUCs can be a manifestation of underlying biological dysfunction, while also leading to biological impacts (generation and exacerbation of health conditions, treatment side effects). NCGUCs confer a large, poorly understood burden to individuals and society. An evidence-base to describe the comprehensive burden is needed. Measurement of NCGUC burdens should incorporate multiple levels of well-being and social ecology, a life course perspective, and potential interactions between NCGUCs and genetics, sex, race, and gender. This approach would elucidate accumulated impacts and potential health inequities in experienced burdens. Uncovering the hidden burden of NCGUCs may draw attention and resources (eg, new research and improved treatments) to this important domain of health.


Comprehensive Geriatric Assessment for Prevention of Delirium After Hip Fracture: A Systematic Review of Randomized Controlled Trials.
To assess the efficacy of comprehensive geriatric assessment (CGA) in prevention of delirium after hip fracture. Systematic review and metaanalysis. Ward based models on geriatrics wards and visiting team based models on orthopaedics wards were included. Four trials (three European, one U.S.; 973 participants) were identified. Two assessed ward-based, and two assessed team-based interventions. MEDLINE, EMBASE, CINAHL and PsycINFO databases; Clinicaltrials.gov; and the Central Register of Controlled Trials were searched. Reference lists from full-text articles were reviewed. Incidence of delirium was the primary outcome. Length of stay, delirium severity, institutionalization, long-term cognition and mortality were predefined secondary outcomes. Duration of delirium was included as a post hoc outcome. There was a significant reduction in delirium overall (relative risk (RR) = 0.81, 95% confidence interval (CI) = 0.69-0.94) in the intervention group. Post hoc subgroup analysis found this effect to be preserved in the team-based intervention group (RR = 0.77, 95% CI = 0.61-0.98) but not the ward-based group. No significant effect was observed on any secondary outcome. There was a reduction in the incidence of delirium after hip fracture with CGA. This is in keeping with results of non-randomized controlled trials and trials in other populations. Team-based interventions appeared superior in contrast to the Ellis CGA paper, but it is likely that heterogeneity in interventions and population studied affected this.


Anion Specific Effects Drive the Formation of Li-Salt Based Aqueous Biphasic Systems.
Aqueous biphasic systems (ABSs) can form when mixing water with two compounds such as polymers, ionic liquids, or simple salts. While this phenomenon has been known for decades and found applications in various fields such as biology, recycling, or even more recently electrochemistry, the physics behind the formation of ABSs remains ill-understood. It was recently demonstrated that ABSs can be composed of two salts sharing the same cation (Li+) but different anions (sulfonamide and halide). Interestingly, their formation could not be explained by the position of the anions within the chaotropic/kosmotropic series and was rather proposed to originate from an anion size mismatch, albeit the size for these anions was never measured yet owing to the lack of a proper experimental methodology. Here, we combine experimental techniques and molecular simulations to assess the specific effects (size, shape, hydrophobic/hydrophilic character) of a series of anions and correlate them with the formation of ABSs. We demonstrate that while the anion size mismatch is a prerequisite for the formation of Li-salts based ABSs, their shape can also play an important role, providing general guidelines for forming new ABSs with potential future applications.


Decreased Symptoms without Augmented Skin Blood Flow in Subjects with RLS/WED after Vibration Treatment.
Vascular disturbances leading to tissue hypoxia have been named as a possible cause for RLS/WED. Vibration to the whole body (WBV) in subjects with RLS/WED results in increased skin blood flow (SBF). The aims of this investigation were to (1) determine if a two-week treatment with WBV will decrease symptoms associated with RLS/WED and, (2) if so, determine if the mechanism for improvement in symptoms is related to an increase in SBF, as measured in flux. Eleven subjects with RLS/WED underwent 2 weeks of 14-minute intermittent WBV and a 2-week sham treatment in randomized order. Pre and post intervention RLS symptom severity were compared. Baseline SBF was compared between subjects with RLS/WED and an age- and sex-matched control group. A crossover design (aim 1) and a matched case-control design and repeated measures design (aim 2) were used. The data analyses consisted of 2-sample and paired t-tests; where applicable we used a standard crossover design adjustment. WBV did significantly decrease symptoms associated with RLS/WED compared to baseline data and compared to sham treatment. The baseline flux was significantly lower in RLS/WED subjects than matched controls, but this deficit was negated with WBV. There was no increase in resting SBF over the 2 weeks of treatment. Subjects with RLS/WED have decreased SBF but are able to increase flux to the same level as normal subjects with WBV. A 2-week intervention with WBV decreases symptoms associated with RLS/WED, but this does not seem to be related to an increase in resting SBF.


Transbronchial lung biopsy via the fiberoptic bronchoscope in the diagnosis of diffuse pulmonary infiltrates in the immunosuppressed host.
The differential diagnosis of diffuse pulmonary infiltrates in the immunosuppressed patient is broad. Many of the etiologies are amenable to specific therapy. Although endobronchial brush biopsy has been used to identify many infections, including Pneumocystis carinii, it was less effective in the diagnosis of noinfectious infiltrates. Transbronchial lung biopsy via the fiberoptic bronchoscope provided lung tissue that could be studied histologically, which increased the diagnostic yield without the morbidity and mortality of open or percutaneous lung biopsies.-


Single-cell analysis of innate spinal cord regeneration identifies intersecting modes of neuronal repair.
Adult zebrafish have an innate ability to recover from severe spinal cord injury. Here, we report a comprehensive single nuclear RNA sequencing atlas that spans 6 weeks of regeneration. We identify cooperative roles for adult neurogenesis and neuronal plasticity during spinal cord repair. Neurogenesis of glutamatergic and GABAergic neurons restores the excitatory/inhibitory balance after injury. In addition, transient populations of injury-responsive neurons (iNeurons) show elevated plasticity between 1 and 3 weeks post-injury. Using cross-species transcriptomics and CRISPR/Cas9 mutagenesis, we found iNeurons are injury-surviving neurons that share transcriptional similarities with a rare population of spontaneously plastic mouse neurons. iNeurons are required for functional recovery and employ vesicular trafficking as an essential mechanism that underlies neuronal plasticity. This study provides a comprehensive resource of the cells and mechanisms that direct spinal cord regeneration and establishes zebrafish as a model of plasticity-driven neural repair.


Body-weight distribution on forelimbs in rat tail-suspension model.
To understand the tail-suspension model to simulate weightlessness better, this study was to investigate the relationship of the amount of body weight supported by forelimbs between the tilt angles of rat in the model. Normal rat had at least two basic postures. One was standing or walking, in which the forelimbs bear 44.6% of the body weight; the other one was resting, in which 23.9% of body weight was placed on the forelimbs. As for tail-suspended rat, body-weight distribution on forelimbs was linearly related to tilt angle. The linear relationship was y = -0.7423x + 70.849, R2 = 0.9269. The tilt angle should be approximately 35 degrees if normal standing load of 44.6% body weight was placed on the forelimbs. On the other hand, it should be approximately 63 degrees if normal resting load of 23.9% of body weight was placed on forelimbs. Furthermore, the body load on forelimbs in tail-suspension model became much larger if the period of different postures was considered. Therefore, it should be careful if forelimbs are used to be as convenient internal control in tail-suspended rats.


Endothelial nitric oxide synthase 27VNTR (4b/4a) gene polymorphism and the risk of diabetic microvascular complications in Chinese populations.
To access the association of endothelial nitric oxide synthase (eNOS) gene 27VNTR (4b/4a) polymorphism with diabetic microvascular complications (DMI) susceptibility in Chinese populations. Data were retrieved in a systematic manner on PubMed database, ISI Web of Knowledge, the Cochrane Library, Chinese National Knowledge Infrastructure, and Wanfang, as well as manual searching of the references of the identified articles. Odds ratio (OR) and 95% confidence interval were used to evaluate the strength of associations. Potential sources of heterogeneity and publication bias were explored. Twelve published articles with thirteen outcomes including 1484 DMI patients and 1225 diabetic controls were included in the meta-analysis. The results showed no evidence for significant association of 27VNTR (4b/4a) polymorphism with DMI risk in dominant model (OR=1.36, 95% CI=0.90-2.06, P=0.15), and similar results were obtained in the allelic, additive and the recessive models (P﹥0.05). Our meta-analysis suggests that eNOS gene 27VNTR (4b/4a) polymorphism might not be a risk factor for DMI in Chinese populations.


Optimal method for reliable lateral spread response monitoring during microvascular decompression surgery for hemifacial spasm.
In this study, we propose an optimal method for monitoring the key electrophysiological sign, the Lateral Spread Response (LSR), during microvascular decompression (MVD) surgery for hemifacial spasm (HFS). Current monitoring methods and interpretations of LSR remain unclear, leading to potential misinterpretations and undesirable outcomes." We prospectively collected data from patients undergoing MVD for HFS, including basic demographics, clinical characteristics, and surgical outcomes. Stimulation intensity was escalated by 1 mA increments to identify the optimal range for effective LSR. We designated the threshold at which we can observe LSR as THR1 and THR2 for when LSR disappears, with high-intensity stimulation (30 mA) designated as THR30. Subsequently, we compared abnormal muscle responses (AMR) between the optimal range (between THR1 and THR2) and THR30. Additionally, we conducted an analysis to identify and assess factors associated with artifacts and their potential impact on clinical outcomes. As stimulation intensity increases, the onset latency to detect AMR was shortened. The first finding of the study was high intensity stimulation caused artifact that mimic the wave of LSR. Those artifacts were observed even after decompression thus interfere interpretation of disappearance of LSR. Analyzing the factors related to the artifact, we found the AMR detected at onset latency below 9.6 ms would be the lateral spreading artifact (LSA) rather than true LSR. To avoid false positive LSR from LSA, we should stepwise increase stimulation intensity and not to surpass the intensity that cause LSR onset latency below 10 ms.


Establishment of a method to rapidly assay bacterial persister metabolism.
Bacterial persisters exhibit an extraordinary tolerance to antibiotics that is dependent on their metabolic state. Although persister metabolism promises to be a rich source of antipersister strategies, there is relatively little known about the metabolism of these rare and transient phenotypic variants. To address this knowledge gap, we explored the use of several techniques, and we found that only one measured persister metabolism. This assay was based on the phenomenon of metabolite-enabled aminoglycoside killing of persisters, and we used it to characterize the metabolic heterogeneity of different persister populations. From these investigations, we determined that glycerol and glucose are the most ubiquitously used carbon sources by various types of Escherichia coli persisters, suggesting that these metabolites might prove beneficial to deliver in conjunction with aminoglycosides for the treatment of chronic and recurrent infections. In addition, we demonstrated that the persister metabolic assay developed here is amenable to high-throughput screening with the use of phenotype arrays.


Targeting the CtBP1-FOXM1 transcriptional complex with small molecules to overcome MDR1-mediated chemoresistance in osteosarcoma cancer stem cells.
Chemoresistance is a major barrier for the chemotherapy of osteosarcoma. The induction of multidrug resistance protein 1 (MDR1), an ATP-dependent transporter, can efflux anti-cancer drugs, thereby decreasing chemosensitivity. However, an actual involvement of MDR1 in the chemoresistance of osteosarcoma cells has not been established. We obtained two cisplatin (CDDP)-resistant osteosarcoma cancer stem cell (CSC) lines using sphere formation medium supplemented with CDDP. These two CDDP-resistant CSC cell lines showed substantial cell proliferation, colony formation, cell invasion, and in vivo tumor growth in the presence of CDDP. Microarray analysis revealed that three genes, MDR1, FOXM1 (forkhead box M1), and CtBP1 (C-Terminal binding protein 1), showed significant overexpression in both cell lines. Mechanistically, CtBP1 assembled with FOXM1 to form a transcriptional complex, which docked onto the MDR1 promoter to activate MDR1 expression. Knockdown or inhibition of the CtBP1-FOXM1 components with specific small molecules, including NSM00158 and NSC95397 for CtBP1 and RCM1 for FOXM1, significantly repressed MDR1 expression. Administration of these three small molecules also significantly inhibited tumor growth in mouse tumor xenograft model. The MDR1-mediated chemoresistance could be reversed by NSM00158 and RCM1. Collectively, our data revealed that the CtBP1-FOXM1 complex activated MDR1 expression and that targeting this complex with their specific inhibitors could reverse MDR1-mediated chemoresistance both in vitro and in vivo. Our results indicate a new therapeutic strategy for overcoming chemoresistance during osteosarcoma treatment.


Age-related differences in white matter integrity and cognitive function are related to APOE status.
While an extensive literature is now available on age-related differences in white matter integrity measured by diffusion MRI, relatively little is known about the relationships between diffusion and cognitive functions in older adults. Even less is known about whether these relationships are influenced by the apolipoprotein (APOE) ε4 allele, despite growing evidence that ε4 increases cognitive impairment in older adults. The purpose of the present study was to examine these relationships in a group of community-dwelling cognitively normal older adults. Data were obtained from a sample of 126 individuals (ages 52-92) that included 32 ε4 heterozygotes, 6 ε4 homozygotes, and 88 noncarriers. Two measures of diffusion, the apparent diffusion coefficient (ADC) and fractional anisotropy (FA), were obtained from six brain regions-frontal white matter, lateral parietal white matter, the centrum semiovale, the genu and splenium of the corpus callosum, and the temporal stem white matter-and were used to predict composite scores of cognitive function in two domains, executive function and memory function. Results indicated that ADC and FA differed with increasing age in all six brain regions, and these differences were significantly greater for ε4 carriers compared to noncarriers. Importantly, after controlling for age, diffusion measures predicted cognitive function in a region-specific way that was also influenced by ε4 status. Regardless of APOE status, frontal ADC and FA independently predicted executive function scores for all participants, while temporal lobe ADC additionally predicted executive function for ε4 carriers but not noncarriers. Memory scores were predicted by temporal lobe ADC but not frontal diffusion for all participants, and this relationship was significantly stronger in ε4 carriers compared to noncarriers. Taken together, age and temporal lobe ADC accounted for a striking 53% of the variance in memory scores within the ε4 carrier group. The results provide further evidence that APOE ε4 has a significant impact on the trajectory of age-related cognitive functioning in older adults. Possible mechanisms are discussed that could account for the associations between ε4, diffusion, and cognitive function, including the influence of ε4 on neural repair, oxidative stress, and the health of myelin-producing oligodendroglia.


Piper methysticum (kava kava).
Piper methysticum (kava kava) is a plant native to the Pacific Island region, and has been used ceremonial for thousands of years. The active ingredients are a group of substances know as kava lactones (AKA kava pyrones). Four lactones in kava have been found to have significant analgesic and anesthetic effects via non-opiate pathways. Kava's most popular application is as a natural anxiolytic, comparing favorably in several studies to a number prescription medications, including benzodiazepines. CNS effects seem to be mediated by several mechanisms. Studies have been conflicting regarding its GABA-receptor-binding capacity, although this has been found to occur in some studies. In vitro kava has been found to block norepinephrine uptake. It also has some anti-convulsant capabilities, which appear to be mediated by Na+ channel receptor sites. The therapeutic dosage is in the range of 50-70 mg kava lactones three times daily. The most common side effect, usually seen only with long-term, heavy usage of the herb, is a scaly skin rash called "kava dermopathy." It has also been know to potentiate other medications such as barbiturates and Xanax.


Long-term exposure to ambient fine particulate pollution induces insulin resistance and mitochondrial alteration in adipose tissue.
We have previously shown that chronic exposure to ambient fine particulate matter (less than 2.5 μm in aerodynamic diameter, PM₂.₅) pollution in conjunction with high-fat diet induces insulin resistance through alterations in inflammatory pathways. In this study, we evaluated the effects of PM₂.₅ exposure over a substantive duration of a rodent's lifespan and focused on the impact of long-term exposure on adipose structure and function. C57BL/6 mice were exposed to PM₂.₅ or filtered air (FA) (6 h/day, 5 days/week) for duration of 10 months in Columbus, OH. At the end of the exposure, PM₂.₅-exposed mice demonstrated insulin resistance (IR) and a decrease in glucose tolerance compared with the FA-exposed group. Although there were no significant differences in circulating cytokines between PM₂.₅- and FA-exposed groups, circulating adiponectin and leptin were significantly decreased in PM₂.₅-exposed group. PM₂.₅ exposure also led to inflammatory response and oxidative stress as evidenced by increase of Nrf2-regulated antioxidant genes. Additionally, PM₂.₅ exposure decreased mitochondrial count in visceral adipose and mitochondrial size in interscapular adipose depots, which were associated with reduction of uncoupling protein 1 (UCP1) expression and downregulation of brown adipocyte-specific gene profiles. These findings suggest that long-term ambient PM₂.₅ exposure induces impaired glucose tolerance, IR, inflammation, and mitochondrial alteration, and thus, it is a risk factor for the development of type 2 diabetes.


[Patient follow up and monitoring: some notes of interest].
The monitoring or follow-up of a person subjected to a health care intervention is a daily routine nursing activity. In the chronic patient the monitoring or follow-up is a core activity and forms a part of the ritual in the periodic visits. In spite of the high percentage workload and use of resources that patient follow up represents within the health system, it is a poorly investigated area. In the present article we examine the differences between a test versus a monitoring strategy, the conditions that justify the monitoring, the methodology to evaluate a follow-up strategy,, as well as creating initiatives to improve monitoring (MaDOx) worldwide.


Analysis of Motor Control and Learning in Human-Robot Interaction during Game Guided Movements.
Robotic devices can be used in upper limb rehabilitation in order to help the total or partial functional recovery. Robots can perform repetitive activities for a long period of time, which may be beneficial for rehabilitation processes. In this context, this study uses a bi-manual robotic device to investigate motor learning and control for the upper limbs among different game guided tasks, and inspect the user's grip force exerted in response to perturbations. The robotic device resembles a bicycle handlebar, instrumented with load cells to measure torques and grip forces. It is equipped with a DC motor to apply external torques to the guiding system. A game was developed containing in-game and physical perturbations to the natural movement of the handlebar. Tests were carried out with 16 healthy subjects that were instructed to move the handlebar guiding a character displayed on the screen with the objective of collecting tokens to get the higher score in the game. During the trials, corresponding data from the game and the load cells were collected and used to infer the learning process, the mean error in the trajectory and the variations in the force applied to the handles of the handlebar. Analyses showed that there was learning in the first repetitions, and the learning was retained further. The higher values of the grip force occurred when there was a physical perturbation to the handlebar's natural movement. The larger errors in the trajectories occurred immediately after the perturbations ended. In conclusion, there was a performance improvement, probably related to learning. The increase of the mean error at the transitions of the perturbations indicates the need for adaptation to the new conditions of the task.


GLI2 cooperates with ZEB1 for transcriptional repression of CDH1 expression in human melanoma cells.
In melanoma cells, high expression of the transcription factor GLI2 is associated with increased invasive potential and loss of E-cadherin expression, an event reminiscent of the epithelial-to-mesenchymal transition (EMT). Herein, we provide evidence that GLI2 represses E-cadherin gene (CDH1) expression in melanoma cells via distinct mechanisms, enhancing transcription of the EMT-activator ZEB1 and cooperative repression of CDH1 gene transcription via direct binding of both GLI2 and ZEB1 to two closely positioned Kruppel-like factor-binding sites within the CDH1 promoter. GLI2 silencing rescued CDH1 expression except in melanoma cell lines in which the CDH1 promoter was hypermethylated. Proximity ligation assays identified GLI2-ZEB1 complexes in melanoma cell nuclei, proportional to endogenous GLI2 and ZEB1 expression, and whose accumulation was enhanced by the classical EMT inducer TGF-β. These data identify GLI2 as a critical modulator of the cadherin switch in melanoma, a molecular process that is critical for metastatic spread of the disease.


Remote teaching system for robotic surgery and its validation: results of a randomized controlled study.
Currently, only a limited number of remote assistance modalities are utilized in the basic phase of robotic surgery training to facilitate the rapid acquisition of robotic surgery skills by surgeons. This study aimed to investigate the benefits of real-time remote surgical robotic skill training based on a multi-channel video recording and playback system. We randomly divided 40 medical students without prior expertise in the use of surgical robots into two groups to assess the performance of trainees on a robotic simulator (Mimic dV-Trainer). The remote group received remote training, while the control group received live one-on-one guidance. We compared the learning curves of the two groups based on simulator scores. Furthermore, the NASA task load index (NASA-TLX) scale was used to measure the fatigue load of the trainers. We observed no significant differences in the demographics or initial baseline skill levels between the two groups. Participants in the remote group achieved higher total scores in the Match Board 2 and Thread the Rings 1 exercises compared to the control group. In addition, trainers in the remote group reported lower subjective fatigue load than in the control group. The remote approach to surgical robotics skills training based on the Remote Teaching System for Robotic Surgery (ReTeRoS) is both feasible and has the potential for large-scale training.


IRAK-M removal counteracts dendritic cell vaccine deficits in migration and longevity.
To function optimally as vaccines, dendritic cells (DCs) must actively migrate to lymphoid organs and maintain a viable, mature state for sufficient time to effectively present their Ag to cognate T cells. Unfortunately, mature DCs rapidly lose viability and function after injection, and only a minority leaves the vaccine site and migrates to lymph nodes. We show that all of these functions can be enhanced in DCs by removal of IL-1R-associated kinase M (IRAK-M). We found that IRAK-M is induced in DCs by TLR ligation and that its absence from these cells leads to increased activation of the p38-MAPK and NF-κB pathways, which, in turn, improves DC migration to lymph nodes, increases their longevity, and augments their secretion of Th1-skewing cytokines and chemokines. These biological effects have immunological consequences. IRAK-M(-/-) DCs increase the proliferation and activation of Ag-specific T cells, and a single vaccination with Ag-pulsed, LPS-matured IRAK-M(-/-) DCs eliminates established tumors and prolongs the survival of EG7 or B16.f10 tumor-bearing mice, without discernible induction of autoimmune disease. Thus, manipulation of IRAK-M levels can increase the potency of DC vaccines by enhancing their Ag-presenting function, migration, and longevity.


Leisure Participation of Autistic Adults: An Ecological Momentary Assessment Feasibility Study.
Autistic adults participate less and express lower satisfaction in leisure activities than nonautistic adults, although literature is limited. The multifaceted nature of leisure participation makes it challenging to measure, with most measures being retrospective. Ecological momentary assessments (EMA) can reduce recall bias. This pilot study assessed the feasibility and acceptability of EMA among autistic adults. Participants (N = 40) were recruited via email and online. After completing a baseline interview, participants were asked to complete a once-daily survey for 30 days, in which they received survey links through a text messaging smartphone app. Surveys asked participants to report whether they participated in any leisure activities during the day, their level of enjoyment, with whom they interacted, and where they participated. The EMA appeared feasible in this sample, as participants completed the daily survey on average 27.05 (SD = 3.92) days. Regarding acceptability, most agreed that survey timing was convenient, that it was easy to enter responses and answer questions daily, and that they had enough response time. Overall, this study supports the use of EMA methodology among autistic adults. Future research should follow and improve upon these EMA data collection practices to examine daily behavior and well-being among autistic adults.


TraC of IncN plasmid pKM101 associates with membranes and extracellular high-molecular-weight structures in Escherichia coli.
Conjugative transfer of IncN plasmid pKM101 is mediated by the TraI-TraII region-encoded transfer machinery components. Similar to the case for the related Agrobacterium tumefaciens T-complex transfer apparatus, this machinery is needed for assembly of pili to initiate cell-to-cell contact preceding DNA transfer. Biochemical and cell biological experiments presented here show extracellular localization of TraC, as suggested by extracellular complementation of TraC-deficient bacteria by helper cells expressing a functional plasmid transfer machinery (S. C. Winans, and G. C. Walker, J. Bacteriol. 161:402-410, 1985). Overexpression of TraC and its export in large amounts into the periplasm of Escherichia coli allowed purification by periplasmic extraction, ammonium sulfate precipitation, and column chromatography. Whereas TraC was soluble in overexpressing strains, it partly associated with the membranes in pKM101-carrying cells, possibly due to protein-protein interactions with other components of the TraI-TraII region-encoded transfer machinery. Membrane association of TraC was reduced in strains carrying pKM101 derivatives with transposon insertions in genes coding for other essential components of the transfer machinery, traM, traB, traD, and traE but not eex, coding for an entry exclusion protein not required for DNA transfer. Cross-linking identified protein-protein interactions of TraC in E. coli carrying pKM101 but not derivatives with transposon insertions in essential tra genes. Interactions with membrane-bound Tra proteins may incorporate TraC into a surface structure, suggested by its removal from the cell by shearing as part of a high-molecular-weight complex. Heterologous expression of TraC in A. tumefaciens partly compensated for the pilus assembly defect in strains deficient for its homolog VirB5, which further supported its role in assembly of conjugative pili. In addition to its association with high-molecular-weight structures, TraC was secreted into the extracellular milieu. Conjugation experiments showed that secreted TraC does not compensate transfer deficiency of TraC-deficient cells, suggesting that extracellular complementation may rely on cell-to-cell transfer of TraC only as part of a bona fide transfer apparatus.


A comparison of risk factors and severity of ischemic stroke in female and male genders in North-West Iran: A cross-sectional study.
Gender difference has been reported in stroke risk factors and disease history. The aim of this study was to compare risk factors and the severity of ischemic stroke based upon modified Rankin Scale (mRS) and hospital mortality between two genders. In a cross-sectional study, 341 patients (44% males and 56% females with a mean age of 68.94 ± 12.74 years) with ischemic stroke, who were hospitalized in the neurology wards of two referral university hospital of North-West Iran (Imam Reza and Razi Hospitals), from the beginning to the end of 2011 were selected and assessed. Gender difference in terms of demographic findings, vascular risk factors, 7(th) day mRS, and hospital mortality (during admission) were evaluated. In 2.6% of cases, mRS was found to be less than 2 (favorable) and in 97.4% of cases; mRS was 2-5 (with disability). No significant difference in ischemic stroke severity based on mRS was observed between two genders. There was a significant difference in the rate of hypertension (females = 72.3%, males = 59.3%, P = 0.010), diabetes (females = 28.8%, males = 18.7%, P = 0.030), smoking (females = 6.3%, males = 35.3%, P < 0.001). No significant difference was seen in other risk factors between two genders. There was no significant difference in the mortality rate, which constituted 8.9% and 4.7% in females and males respectively (P = 0.140). The evidence from the present study suggests that despite the existence of some difference between risk-factors in two genders, there was no difference in terms of ischemic stroke severity and mortality rate between two genders.


High duty cycle echolocation and prey detection by bats.
There are two very different approaches to laryngeal echolocation in bats. Although most bats separate pulse and echo in time by signalling at low duty cycles (LDCs), almost 20% of species produce calls at high duty cycles (HDCs) and separate pulse and echo in frequency. HDC echolocators are sensitive to Doppler shifts. HDC echolocation is well suited to detecting fluttering targets such as flying insects against a cluttered background. We used two complementary experiments to evaluate the relative effectiveness of LDC and HDC echolocation for detecting fluttering prey. We measured echoes from fluttering targets by broadcasting artificial bat calls, and found that echo amplitude was greatest for sounds similar to those used in HDC echolocation. We also collected field recordings of syntopic LDC and HDC bats approaching an insect-like fluttering target and found that HDC bats approached the target more often (18.6% of passes) than LDC bats (1.2% of passes). Our results suggest that some echolocation call characteristics, particularly duty cycle and pulse duration, translate into improved ability to detect fluttering targets in clutter, and that HDC echolocation confers a superior ability to detect fluttering prey in the forest understory compared with LDC echolocation. The prevalence of moths in the diets of HDC bats, which is often used as support for the allotonic frequency hypothesis, can therefore be partly explained by the better flutter detection ability of HDC bats.


[Negative mental disorders in epilepsy].
Objective. To search for clinical features of negative mental disorders in epilepsy. Material and methods. We studied 78 patients, aged from 18 to 40 years,, including 43 patients diagnosed with epilepsy for the first time (main group) and 35 patients with the previous diagnosis of epilepsy (comparison group). Diagnosis was made according to ICD-10. Results. We observed a broad spectrum of persistent mental disorders, including negative disorders. Personality disorders and cognitive deficit were most significant negative disorders. Personality disorders included two opposite types: explosive-epileptoid features with paranoiac signs and defensive disorder. Inertia, slow mobility, slowness of the rate of thought processes, the domination of egocentric, perseverative, visual-image thinking were characteristic of thought disorders. Conclusion. The authors suggest the systematic of these disorders.


[Age-related aspects of depressive states].
The significant variety of psychopathological presentations of depression causes difficulties in its diagnosis. Depression of middle age is subdivided depending on the predomination of one or another disorder in the clinical picture. There are simple depression and complicated depression with additional psychopathological symptoms. The most difficult is to diagnose depression with onset in early childhood, including infant's age, when the psychopathological presentations depend on the consequences of the perinatal CNS lesion, maternal deprivation and other factors. In children, the clinical picture of depression differs by syndromologic incompleteness, peculiarity of phenomenological presentations and polymorphism of disorders. In adolescence, depression is characterized by the predominance of behavioral disorders, marked psychopathic-like disorders, suicidal attempts, higher frequency of bipolar forms. Depression with late age at onset differs by the abundance of somatic and hypochondriac complaints, anxiety and depressive delusion accompanied by the transitory aggravation of cognitive impairment (depressive pseudodementia).


Water security in the Canadian Prairies: science and management challenges.
In this paper, we discuss the multiple dimensions of water security and define a set of thematic challenges for science, policy and governance, based around cross-scale dynamics, complexity and uncertainty. A case study of the Saskatchewan River basin (SRB) in western Canada is presented, which encompasses many of the water-security challenges faced worldwide. A science agenda is defined based on the development of the SRB as a large-scale observatory to develop the underpinning science and social science needed to improve our understanding of water futures under societal and environmental change. We argue that non-stationarity poses profound challenges for existing science and that new integration of the natural sciences, engineering and social sciences is needed to address decision making under deep uncertainty. We suggest that vulnerability analysis can be combined with scenario-based modelling to address issues of water security and that knowledge translation should be coupled with place-based modelling, adaptive governance and social learning to address the complexity uncertainty and scale dynamics of contemporary water problems.


Microplastics in brown trout (Salmo trutta Linnaeus, 1758) from an Irish riverine system.
Rivers play an important role in the overall transport of microplastic pollution (1 μm to 5 mm), with fluvial dynamics expected to influence biotic interactions, particularly for fish. So far, there have been few assessments of microplastics in freshwater salmonids. The prevalence (i.e. percentage occurrence) and burden (i.e. abundance per fish) of microplastics were assessed in the gastrointestinal tracts (GITs) and stomach contents (SCs) of 58 brown trout Salmo trutta Linnaeus, 1758 sampled at six sites along the River Slaney catchment in south-east Ireland. Sites were divided into two classifications (high and low exposure) based on proximity to microplastic pollution sources, comprising three sites each. Analysis of biological traits (e.g. fish length) and diet was performed on the same fish to determine possible factors explaining microplastic burden. Microplastics were found in 72% of fish having been recovered from 66% of GITs (1.88 ± 1.53 MPs fish⁻1) and 28% of SCs (1.31 ± 0.48 MPs fish⁻1). Fibres were the dominant particle type recovered from GITs (67%) and SCs (57%) followed by fragments. No difference in median microplastic burden was observed between fish collected in high and low exposure sites. Microplastic burden was unrelated to fish fork length, while microplastic size distribution (100 ≤ 350 μm, 350 μm to ≤ 5 mm) was unrelated to S. trutta age class estimates. Furthermore, microplastic burden was not explained by dietary intake. Though further research is necessary, this study showed the presence of microplastics in wild S. trutta collected from an Irish riverine system, which could have further implications for top-level consumers that feed on the species, including humans. Further analysis is required to determine possible trophic linkages for the species, with respect to microplastics, and to assess the suitability of S. trutta for monitoring microplastics in river systems.


African Swine Fever: Disease Dynamics in Wild Boar Experimentally Infected with ASFV Isolates Belonging to Genotype I and II.
After the re-introduction of African swine fever virus (ASFV) genotype II isolates into Georgia in 2007, the disease spread from Eastern to Western Europe and then jumped first up to Mongolian borders and later into China in August 2018, spreading out of control and reaching different countries of Southeast Asia in 2019. From the initial incursion, along with domestic pigs, wild boar displayed a high susceptibility to ASFV and disease development. The disease established self-sustaining cycles within the wild boar population, a key fact that helped its spread and that pointed to the wild boar population as a substantial reservoir in Europe and probably also in Asia, which may hinder eradication and serve as the source for further geographic expansion. The present review gathers the most relevant information available regarding infection dynamics, disease pathogenesis and immune response that experimental infections with different ASFV isolates belonging to genotype I and II in wild boar and feral pigs have generated. Knowledge gaps in areas such as disease pathogenesis and immune response highlights the importance of focusing future studies on unravelling the early mechanisms of virus-cell interaction and innate and/or adaptive immune responses, knowledge that will contribute to the development of efficacious treatments/vaccines against ASFV.


The health benefits of wine.
Epidemiologic studies from numerous disparate populations reveal that individuals with the habit of daily moderate wine consumption enjoy significant reductions in all-cause and particularly cardiovascular mortality when compared with individuals who abstain or who drink alcohol to excess. Researchers are working to explain this observation in molecular and nutritional terms. Moderate ethanol intake from any type of beverage improves lipoprotein metabolism and lowers cardiovascular mortality risk. The question now is whether wine, particularly red wine with its abundant content of phenolic acids and polyphenols, confers additional health benefits. Discovering the nutritional properties of wine is a challenging task, which requires that the biological actions and bioavailability of the >200 individual phenolic compounds be documented and interpreted within the societal factors that stratify wine consumption and the myriad effects of alcohol alone. Further challenge arises because the health benefits of wine address the prevention of slowly developing diseases for which validated biomarkers are rare. Thus, although the benefits of the polyphenols from fruits and vegetables are increasingly accepted, consensus on wine is developing more slowly. Scientific research has demonstrated that the molecules present in grapes and in wine alter cellular metabolism and signaling, which is consistent mechanistically with reducing arterial disease. Future research must address specific mechanisms both of alcohol and of polyphenolic action and develop biomarkers of their role in disease prevention in individuals.


[Bilateral peripheral facial paralysis and Guillain-Barré syndrome].
The Guillain-Barrè syndrome is a polyneuropathy of acute onset that initially tends to produce motor damage of the lower limbs, albumin-cytological dissociation in cerebrospinal fluid and electrophysiological findings suggestive of demyelination. This entity produces a wide variety of symptoms, including damage to the facial nerve in as many as half of all episodes, which means that this syndrome should be considered in the differential diagnosis of facial palsy, particularly bilateral and synchronous cases. In a review of six cases of Guillain-Barrè syndrome in which bilateral facial palsy occurred at some point, a history of immediate infection was confirmed in five, mainly related to Herpesvirinae and Campylobacter. One case began with bilateral palsy, whereas palsy appeared almost simultaneously with the rest of the symptoms in the other cases. High protein levels without cells in cerebrospinal fluid and electrophysiological patterns of slow facial nerve conduction were confirmed in every patient. Treatment with intravenous immunoglobulins resolved all episodes in less than six months. We conclude by reiterating the need for awareness of this syndrome in every case of peripheral VIIth nerve palsy, especially bilateral cases, although the associated symptoms were not very useful, if at all useful.


Work Patterns and Intensity of Chinese Surgical Residents- A Multicenter Time-and-Motion Study.
This study aimed to record and analyze surgical resident trainee time allocation among junior doctors in China in order to understand the training environment and optimize realistic training and patient care objectives. Multicenter observational time and motion study. Multicenter, carried out in 5 tier 3 public hospitals in 5 provinces across China. Surgical resident trainees at various stages of training were eligible to enter the study, total n = 44. Registered nurses were eligible to be observers, n = 4 from each hospital. An expert team comprising 4 chief surgeons and 10 surgical residents participated in establishing the clinical activity list. Participants were observed during working hours (08.00-17.00) for 10 consecutive working days and time spent on different activities were recorded. Work patterns between hospitals were often dissimilar. Most time was spent on direct patient care (34.1%; 95% CI, 28.0%-40.1%) followed by indirect patient care (24.4%; 95% CI, 15.5%-33.2%), scholarly activity (21.1%; 95% CI, 13.7%-28.5%) and other (20.4%; 95% CI, 14.1%-26.8%). Subcategory analysis showed that the amount of time spent each day performing certain tasks was 137 minutes for operating theatre tasks, 103 minutes for medical record-keeping, 25 minutes for direct patient contact, 20 minutes being taught, 12 minutes teaching others, 12 minutes hand-over time, and 0 minutes of outpatient clinic attendance. Inter-observer reliability of 96.5% was obtained prior to recordings. Chinese surgical resident work patterns fall within the range found in other international studies albeit with some exceptions. The training environment appears broadly suitable for competence-based surgical training in China. Inadequate outpatient activity has led to changes in trainee work rosters and trainer requirements. Both strengths and deficiencies were confirmed and addressed. Further audit is required.


Changes in hepatic bile secretion following cholecystectomy.
The volume and compositon of hepatic bile was studied in anesthetized dogs before and approximately 18 weeks after cholecystectomy. Dogs had common duct cannulation, cystic duct ligation, and temporary occlusion of the pylorus. An intravenous infusion of sodium taurocholate, 9 muEq per minute, was given throughout the experiment. Bile volume, electrolyte composition, bile salt output, and 14C-mannitol clearance were determined during infusion of sodium taurocholate alone and after addition of secretin, 4 U. per kilogram per hour, to the infusion. Cholecystectomy then was done and the same experiment was repeated 18 weeks later. Cholecystectomy significantly increased bile volume, sodium and chloride output, and 14C-mannitol clearance both in response to taurocholate alone and to taurocholate plus secretin. Bile salt output and bicarbonate output were unchanged. The study shows that cholecystectomy significantly alters the volume and composition of hepatic bile. The data indicate that cholecystectomy does not affect ductular function or hepatic secretion of bile salts but that canalicular function is altered significantly.


DYRK1A interacts with the REST/NRSF-SWI/SNF chromatin remodelling complex to deregulate gene clusters involved in the neuronal phenotypic traits of Down syndrome.
The molecular mechanisms that lead to the cognitive defects characteristic of Down syndrome (DS), the most frequent cause of mental retardation, have remained elusive. Here we use a transgenic DS mouse model (152F7 line) to show that DYRK1A gene dosage imbalance deregulates chromosomal clusters of genes located near neuron-restrictive silencer factor (REST/NRSF) binding sites. We found that Dyrk1a binds the SWI/SNF complex known to interact with REST/NRSF. The mutation of a REST/NRSF binding site in the promoter of the REST/NRSF target gene L1cam modifies the transcriptional effect of Dyrk1a-dosage imbalance on L1cam. Dyrk1a dosage imbalance perturbs Rest/Nrsf levels with decreased Rest/Nrsf expression in embryonic neurons and increased expression in adult neurons. Using transcriptome analysis of embryonic brain subregions of transgenic 152F7 mouse line, we identified a coordinated deregulation of multiple genes that are responsible for dendritic growth impairment present in DS. Similarly, Dyrk1a overexpression in primary mouse cortical neurons induced severe reduction of the dendritic growth and dendritic complexity. We propose that DYRK1A overexpression-related neuronal gene deregulation via disturbance of REST/NRSF levels, and the REST/NRSF-SWI/SNF chromatin remodelling complex, significantly contributes to the neural phenotypic changes that characterize DS.


Opportunities for treatment of the hepatitis C virus-infected patient with chronic kidney disease.
The prevalence of hepatitis C virus (HCV) infection amongst patients with chronic kidney disease (CKD) and end-stage renal disease exceeds that of the general population. In addition to predisposing to the development of cirrhosis and hepatocellular carcinoma, infection with HCV has been associated with extra-hepatic complications including CKD, proteinuria, glomerulonephritis, cryoglobulinemia, increased cardiovascular risk, insulin resistance, and lymphoma. With these associated morbidities, infection with HCV is not unexpectedly accompanied by an increase in mortality in the general population as well as in patients with kidney disease. Advances in the understanding of the HCV genome have resulted in the development of direct-acting antiviral agents that can achieve much higher sustained virologic response rates than previous interferon-based protocols. The direct acting antivirals have either primarily hepatic or renal metabolism and excretion pathways. This information is particularly relevant when considering treatment in patients with reduced kidney function. In this context, some of these agents are not recommended for use in patients with a glomerular filtration rate < 30 mL/min per 1.73 m2. There are now Food and Drug Administration approved direct acting antiviral agents for the treatment of patients with kidney disease and reduced function. These agents have been demonstrated to be effective with sustained viral response rates comparable to the general population with good safety profiles. A disease that was only recently considered to be very challenging to treat in patients with kidney dysfunction is now curable with these medications.


Workplace bullying: concerns for nurse leaders.
The aim of this study was to describe nurses' experiences with and characteristics related to workplace bullying. Although the concept of workplace bullying is gaining attention, few studies have examined workplace bullying among nurses. This was a descriptive study using a convenience sample of 249 members of the Washington State Emergency Nurses Association. The Negative Acts Questionnaire-Revised was used to measure workplace bullying. Of the sample, 27.3% had experienced workplace bullying in the last 6 months. Most respondents who had been bullied stated that they were bullied by their managers/directors or charge nurses. Workplace bullying was significantly associated with intent to leave one's current job and nursing. In seeking remedies to the problem of workplace bullying, nurse leaders need to focus on why this bullying occurs and on ways to reduce its occurrence. This is a critical issue, since it is linked with nurse attrition.


Digitalis.
Cardiac glycosides have played a prominent role in the therapy of congestive heart failure since William Withering codified their use in his late 18th century monograph on the efficacy of the leaves of the common foxglove plant (Digitalis purpurea). Despite their widespread acceptance into medical practice in the ensuing 200 years, both the efficacy and the safety of this class of drugs continue to be a topic of debate. Moreover, despite the fact that the molecular target for the cardiac glycosides, the alpha-subunit of sarcolemmal Na+K+-ATPase (or sodium pump) found on most eukaryotic cell membranes, has been known for several decades, it remains controversial whether the sympatholytic or positive inotropic effects of these agents is the mechanism most relevant to relief of heart failure symptoms in humans with systolic ventricular dysfunction. Herein, we review the molecular and clinical pharmacology of this venerable class of drugs, as well as the manifestations of digitalis toxicity and their treatment. We also review in some detail recent clinical trials designed to examine the efficacy of these drugs in heart failure, with a focus on the Digoxin Investigation Group data set. Although, in our opinion, the data on balance warrant the continued use of these drugs for the treatment of symptoms of heart failure in patients already receiving contemporary multidrug therapy for this disease, the use of digitalis preparations will inevitably decline with the maturation of newer pharmacotherapies.


Simultaneous visualization of seven different DNA probes by in situ hybridization using combinatorial fluorescence and digital imaging microscopy.
Combinatorial labeling of probes (i.e., with two or more different reporters) increases the number of target sequences that can be detected simultaneously by fluorescence in situ hybridization. We have used an epifluorescence microscope equipped with a digital imaging camera and computer software for pseudocoloring and merging images to distinguish up to seven different probes using only three fluorochromes. Chromosome-specific centromere repeat clones and chromosome-specific "composite" probe sets were generated by PCR in which different mixtures of modified nucleotides, including fluorescein-conjugated dUTP, were incorporated. Cosmid clones were labeled similarly by nick-translation. The technique has been used to delineate the centromeres of seven different human chromosomes, on both 4',6-diamidino-2-phenylindole-stained metaphase spreads and interphase nuclei, to map six cosmid clones in a single hybridization experiment and to detect chromosome translocations by chromosome painting. Multiparameter hybridization analysis should facilitate molecular cytogenetics, probe-based pathogen diagnosis, and gene mapping studies.


Effect of Vandetanib on Lung Tumorigenesis in Transgenic Mice Carrying an Activating Egfr Gene Mutation.
Vandetanib (ZactimaTM) is a novel, orally available inhibitor of both vascular endothelial growth factor receptor-2 (VEGFR-2) and epidermal growth factor receptor (EGFR) tyrosine kinase. In the present study, a line of transgenic mice with a mouse Egfr gene mutation (delE748-A752) corresponding to a human EGFR mutation (delE746-A750) was established. The transgenic mice developed atypical adenomatous hyperplasia to adenocarcinoma of the lung at around 5 weeks of age and died of lung tumors at approximately 17 weeks of age. In the mice treated with vandetanib (6mg/kg/day), these lung tumors disappeared and the phosphorylations of EGFR and VEGFR-2 were reduced in lung tissues to levels comparable to those of non-transgenic control mice. The median overall survival time of the transgenic mice was 28 weeks in the vandetanib-treated group and 17 weeks in the vehicle-treated group. Vandetanib significantly prolonged the survival of the transgenic mice (log-rank test, p< 0.01); resistance to vandetanib occurred at 20 weeks of age and the animals died from their lung tumors at about 28 weeks of age. These data suggest that vandetanib could suppress the progression of tumors harboring an activating EGFR mutation.


Custody and access issues with children whose parents are separated or divorced.
This article discusses the effects of family breakdown--and the resulting issues of custody and access--on children. It explores trends in child custody and access and issues surrounding those trends, particularly in terms of the benefits and limitations of joint custody. It then identifies the contextual problems of family breakups (including the relative poverty experienced by mother-led families, the unreliability of financial support from fathers, the complications for children which result from step-parent figures, including serial partners of their parents, the implications of interparental conflict, the need to fully address suspicions and/or allegations of abuse by one parent against another, and the problem of Parental Alienation Syndrome). Typical responses of children to family breakup are then considered, including feelings of loss, guilt, and responsibility, and a sense of divided loyalties--particularly in light of intense or violent parental conflict. And finally, recommendations are made to minimize the detrimental effects of family breakup on children: (a) allowing children choice and flexibility, (b) exploring the benefits of mediation for families, (c) promoting parental co-operation, and (d) encouraging an ongoing relationship with the nonresidential parent.


Identification of helicobacter species in human liver samples from patients with primary hepatocellular carcinoma.
Several studies have shown the presence of helicobacter species in the human biliary tract and in the intestinal tract of animals. Experimental infection by Helicobacter hepaticus in mice causes chronic hepatitis and hepatocellular carcinoma (HCC). This study investigated whether helicobacter species could be detected in the liver of patients with HCC. Liver samples from 20 patients with primary liver carcinoma diagnosed by histopathology and 16 controls without primary liver carcinoma were studied. Histology with standard and immunohistochemical stains, culture, and polymerase chain reaction (PCR) amplification using helicobacter genus specific 16S rRNA primers were used to detect the presence of bacteria. Amplified products were identified by Southern hybridisation and sequencing. A search for other genes specific for Helicobacter pylori was also performed. Helicobacter species 16S rDNA was found in eight of 20 samples of primary liver carcinoma, whereas none of the controls harboured this rDNA. Six helicobacter specific PCR amplicons were sequenced and were found to have 98.5-99.0% similarity to the 16S rDNA of H pylori. Of the eight positive samples, seven were positive in PCR using 26 kDa protein primers and six showed morphological and immunohistochemical evidence of H pylori. The cagA and glmM genes were detected in only two samples. The vacA and rps4 genes were not detected. Helicobacter can be present in the liver of patients with primary liver carcinoma and is probably linked to the carcinogenic process in the liver.


Subchronic and developmental toxicity studies of n-butyl propionate vapor in rats.
Two inhalation studies were conducted to evaluate the possible subchronic and developmental toxic effects of n-butyl propionate. In the subchronic study, Sprague-Dawley rats (15/sex/group) were exposed to 0, 250, 750, or 1500 ppm vapor for 6 h/d, 5 d/wk for 13 wk. Five of the rats per sex per group were held after the final exposure for an 8-wk recovery period. Standard parameters of subchronic toxicity were measured throughout the study, and at the end of exposure and recovery periods, necropsies were performed, organs weighed, and tissues processed for microscopic examination. Exposure did not produce marked treatment-related deaths or adversely affect clinical signs, hematology, clinical chemistries, organ weights, or the histology of major visceral organs. The only systemic toxic effects were significant decreases in body weight, body weight gain, and feed consumption that occurred in 1500 ppm group rats. Morphologic changes were limited to the nasal cavity as evidenced by a concentration-related increased incidence and severity of olfactory epithelium degeneration in rats of the 750 and 1500 ppm groups. These degenerative microscopic alterations were primarily confined to the olfactory epithelium within the dorsal portion of the medial meatus, with lesser involvement of the olfactory mucosae overlying the tips of some of the adjacent ethmoturbinates. Both the systemic and nasal cavity effects appeared reversible after exposure ceased. In the developmental toxicity study, pregnant Sprague-Dawley rats (24/group) were exposed to 0, 500, 1000, or 2000 ppm vapor for 6 h/d on gestation d 6-15 and sacrificed on gestation d 20. All treatment-group dams exhibited significant reductions in body weight, body weight gain, and feed consumption. Gestational parameters were equivalent across all groups and there were no treatment-related developmental or teratogenic effects. The no-observed-adverse effects levels (NOAELs) determined for nbutyl propionate were 250 ppm for subchronic toxicity (based on the olfactory epithelium degeneration) and 22000 ppm for developmental toxicity (no developmental effects at top dose tested). Under the conditions of this study, a NOAEL was not determined for maternal toxicity.


Misinformation about COVID-19: evidence for differential latent profiles and a strong association with trust in science.
The global spread of coronavirus disease 2019 (COVID-19) has been mirrored by diffusion of misinformation and conspiracy theories about its origins (such as 5G cellular networks) and the motivations of preventive measures like vaccination, social distancing, and face masks (for example, as a political ploy). These beliefs have resulted in substantive, negative real-world outcomes but remain largely unstudied. This was a cross-sectional, online survey (n=660). Participants were asked about the believability of five selected COVID-19 narratives, their political orientation, their religious commitment, and their trust in science (a 21-item scale), along with sociodemographic items. Data were assessed descriptively, then latent profile analysis was used to identify subgroups with similar believability profiles. Bivariate (ANOVA) analyses were run, then multivariable, multivariate logistic regression was used to identify factors associated with membership in specific COVID-19 narrative believability profiles. For the full sample, believability of the narratives varied, from a low of 1.94 (SD=1.72) for the 5G narrative to a high of 5.56 (SD=1.64) for the zoonotic (scientific consensus) narrative. Four distinct belief profiles emerged, with the preponderance (70%) of the sample falling into Profile 1, which believed the scientifically accepted narrative (zoonotic origin) but not the misinformed or conspiratorial narratives. Other profiles did not disbelieve the zoonotic explanation, but rather believed additional misinformation to varying degrees. Controlling for sociodemographics, political orientation and religious commitment were marginally, and typically non-significantly, associated with COVID-19 belief profile membership. However, trust in science was a strong, significant predictor of profile membership, with lower trust being substantively associated with belonging to Profiles 2 through 4. Belief in misinformation or conspiratorial narratives may not be mutually exclusive from belief in the narrative reflecting scientific consensus; that is, profiles were distinguished not by belief in the zoonotic narrative, but rather by concomitant belief or disbelief in additional narratives. Additional, renewed dissemination of scientifically accepted narratives may not attenuate belief in misinformation. However, prophylaxis of COVID-19 misinformation might be achieved by taking concrete steps to improve trust in science and scientists, such as building understanding of the scientific process and supporting open science initiatives.


Mediastinal lymphomas: primary mediastinal (thymic) large B-cell lymphoma versus classical Hodgkin lymphoma, histopathologic dilemma solved?
Primary Mediastinal (Thymic) Large B-cell Lymphoma (PMLBL) frequently demonstrates clinical, morphologic and/or immunohistochemical (IHC) features that overlap with mediastinal-classical Hodgkin lymphoma (cHL). Our study compared the immunohistochemical profile of PMLBL versus mediastinal cHL in a cohort of 18 patients, 11 of whom were diagnosed as having PMLBL, and 7 were diagnosed as having mediastinal cHL. Biopsy slides and blocks of all patients were retrieved, and further immunohistochemical workup was done on all the cases. This included staining with a panel of antibodies, including LCA, CD(20), CD(3), CD(30), CD(15), and IRF4/MUM1. All PMLBL cases (100%) showed positivity both for LCA and Pan B (CD(20)), and negativity for Pan T (CD(3)). All mediastinal cHL cases (100%) showed negativity both for LCA and Pan T (CD(3)). Both entities showed variable positivity for IRF4/MUM1, i.e., 73% in PMLBL versus 100% in cHL. CD(30) was expressed in 36% cases of PMLBL versus 100% cases of cHL. CD(15) was expressed in a single case of PMLBL versus 86% cases of cHL. We conclude that 'Leucocyte Cell Antigen' (LCA) is the only marker that was consistently positive in PMLBL and negative in cHL. Other IHC markers, though still helpful, may be expressed in either entity.


Longitudinal determination of cerebral blood flow velocity in neonates with the Doppler technique.
Using the Doppler technique, we followed the postnatal changes of the cerebral blood flow velocity longitudinally. Pulsatility index (PI) was measured in 91 neonates. In 26 normal term infants who were supine, PIs were initially raised (PI = 0.91 +/- 0.10), gradually falling over the first 12 hours after birth and remaining stable thereafter (PI = 0.78 +/- 0.05). PIs tended to be lower in neonates born by caesarean section but remained grossly within the normal range. Low birthweight infants showed high PIs during the first 12 hours after birth. Infants with a prone position showed a narrow standard deviation in PI soon after birth. There was a negative correlation between PI and the conceptional age and the coefficient of the correlation was 0.97156. The stage of active sleep, especially the REM period, was found to be an important factor which decreased the PI. We concluded that neonatal cerebral circulation significantly changes during the perinatal period and is affected by various physiological factors, especially sleep stages, the position of the infant and the conceptional age.


The type IV pilus protein PilU functions as a PilT-dependent retraction ATPase.
Type IV pili are dynamic cell surface appendages found throughout the bacteria. The ability of these structures to undergo repetitive cycles of extension and retraction underpins their crucial roles in adhesion, motility and natural competence for transformation. In the best-studied systems a dedicated retraction ATPase PilT powers pilus retraction. Curiously, a second presumed retraction ATPase PilU is often encoded immediately downstream of pilT. However, despite the presence of two potential retraction ATPases, pilT deletions lead to a total loss of pilus function, raising the question of why PilU fails to take over. Here, using the DNA-uptake pilus and mannose-sensitive haemagglutinin (MSHA) pilus of Vibrio cholerae as model systems, we show that inactivated PilT variants, defective for either ATP-binding or hydrolysis, have unexpected intermediate phenotypes that are PilU-dependent. In addition to demonstrating that PilU can function as a bona fide retraction ATPase, we go on to make the surprising discovery that PilU functions exclusively in a PilT-dependent manner and identify a naturally occurring pandemic V. cholerae PilT variant that renders PilU essential for pilus function. Finally, we show that Pseudomonas aeruginosa PilU also functions as a PilT-dependent retraction ATPase, providing evidence that the functional coupling between PilT and PilU could be a widespread mechanism for optimal pilus retraction.


Prenatal exposure to a maternal low protein diet shortens life span in rats.
Postweaning diet restriction is associated with prolongation of life span, reduced age-related disease and slower ageing. The effects of diet restriction imposed prior to weaning have not been so well characterised, but studies suggest an opposite effect with increased age-related diseases occurring in offspring exposed to undernutrition in prenatal life. It remains unclear whether life span is similarly adversely affected by early diet restriction. The present study in rats aimed to evaluate the impact of a maternal low protein diet upon the life span of the resulting offspring. Rat dams were fed either a 180-gram casein/kg control diet or a 90-gram casein/kg low protein diet from conception until the end of pregnancy. The offspring were then maintained with minimal handling until death from natural causes or distress-necessitated euthanasia. The average life span of female rats exposed to low protein diets in utero was reduced by 11% (p = 0.044, Kaplan-Meier analysis). There was a similar but non- significant trend in the male offspring (control 76 +/- 3 weeks, low protein 73 +/- 3 weeks). In addition the rats exposed to a prenatal low protein diet had significantly higher systolic blood pressure at 4 weeks of age and tended to be smaller than control animals in postnatal life. The results suggest that intrauterine diet restriction reduces life span in rats and contrasts with the well-recognised increase in life span produced by postweaning diet restriction. The timing of the nutritional intervention appears to be critical and recognition of this is relevant to understanding the mechanisms underlying the effects of diet restriction on ageing and life span.


Low frequency of cytotoxic T lymphocytes against the novel HLA-A*0201-restricted JC virus epitope VP1(p36) in patients with proven or possible progressive multifocal leukoencephalopathy.
JC virus (JCV)-specific cytotoxic T lymphocytes (CTL) in peripheral blood are associated with a favorable outcome in patients with progressive multifocal leukoencephalopathy (PML). However, the frequency of these cells in the peripheral blood mononuclear cells (PBMC) of PML patients is unknown. To develop a highly sensitive assay for detecting the cellular immune response against this virus, we performed a CTL epitope mapping study of JCV VP1 major capsid protein by using overlapping peptides. A novel HLA-A*0201-restricted epitope, the VP1(p36) peptide SITEVECFL, was characterized. The cellular immune response against JCV was assessed in 32 study subjects. By combining the results of the (51)Cr release assay on pooled peptides and staining with the HLA-A*0201/JCV VP1(p36) tetramer, VP1-specific CTL were detected in 10 of 11 PML survivors (91%) versus only 1 of 11 PML progressors (9%, P = 0.0003). VP1-specific CTL were also detected in two of two patients recently diagnosed with PML and in four of four human immunodeficiency virus-positive patients with possible PML. The frequency of CTL specific for the novel VP1(p36) and the previously described VP1(p100) epitopes was determined. In two patients, the frequency of CTL specific for the VP1(p36) or VP1(p100) epitopes, as determined by fresh blood tetramer staining (FBTS), ranged from 1/6,000 to 1/24,000 PBMC. A CTL sorting technique combining tetramer staining and selection with immunomagnetic beads allowed the detection of epitope-specific CTL in two cases that were determined to be negative by FBTS. The phenotype of these CTL in vivo was consistent with activated memory cells. These data suggest that, although present in low numbers, JCV-specific CTL may be of central importance in the containment of JCV spread in immunosuppressed individuals.


Hodgkin's disease: complications of therapy and excess mortality.
The long-term survival of patients treated for Hodgkin's disease permits careful evaluation of long-term complications and excess mortality. Between 1960 and 1995, 2498 patients who were treated for Hodgkin's disease at Stanford University were evaluated. Survival, freedom from relapse, and important complications of therapy (cardiac disease and secondary cancers) were analyzed, and risk of mortality from all causes was calculated utilizing absolute excess risk calculations. The risk of death from Hodgkin's disease is 17% at 15 years of follow-up and increases only slightly thereafter. The risk of death from other causes is also 17% at 15 years, but increases sharply thereafter. The major causes of mortality (other than Hodgkin's disease) are secondary cancers and cardiac disease. Second cancers with significant increase in risk include leukemia (acute nonlymphocytic), non-Hodgkin's lymphoma, lung/pleural cancer, breast cancer, melanoma, soft tissue and bone sarcomas, stomach cancer, salivary gland tumors, thyroid cancer, and pancreatic cancer. The absolute excess risk of death from causes other than Hodgkin's disease increases during each five-year follow-up interval for at least 25 years. However, the absolute excess risk of death during similar follow-up periods is less for patients treated in more recent years (1980-1995) than in the prior treatment era (1962-1980). Mortality for causes other than Hodgkin's disease is important in the long-term follow-up of patients. Causes of death are often treatment related. Changes in treatment programs can reduce the long-term excess risk of death from complications of therapy.


Beneficial autoimmunity improves cancer prognosis.
Many tumour antigens that do not arise from cancer cell-specific mutations are targets of humoral and cellular immunity despite their expression on non-malignant cells. Thus, in addition to the expected ability to detect mutations and stress-associated shifts in the immunoproteome and immunopeptidome (the sum of MHC class I-bound peptides) unique to malignant cells, the immune system also recognizes antigens expressed in non-malignant cells, which can result in autoimmune reactions against non-malignant cells from the tissue of origin. These autoimmune manifestations include, among others, vitiligo, thyroiditis and paraneoplastic syndromes, concurrent with melanoma, thyroid cancer and non-small-cell lung cancer, respectively. Importantly, despite the undesirable effects of these symptoms, such events can have prognostic value and correlate with favourable disease outcomes, suggesting 'beneficial autoimmunity'. Similarly, the occurrence of dermal and endocrine autoimmune adverse events in patients receiving immune-checkpoint inhibitors can have a positive predictive value for therapeutic outcomes. Neoplasias derived from stem cells deemed 'not essential' for survival (such as melanocytes, thyroid cells and most cells in sex-specific organs) have a particularly good prognosis, perhaps because the host can tolerate autoimmune reactions that destroy tumour cells at some cost to non-malignant tissues. In this Perspective, we discuss examples of spontaneous as well as therapy-induced autoimmunity that correlate with favourable disease outcomes and make a strong case in favour of this 'beneficial autoimmunity' being important not only in patients with advanced-stage disease but also in cancer immunosurveillance.


Antiglycan antibodies as serological markers in the differential diagnosis of inflammatory bowel disease.
The objective of the study was to evaluate the diagnostic accuracy of recently developed antiglycan serological tests in clinical practice for the diagnosis of Crohn's disease. This study was a cohort analysis of both clinical and biochemical parameters of patients with diagnosed inflammatory bowel disease compared with those in a control population. Antiglycan antibodies were determined using commercially available enzyme immunoassays. The setting was the outpatient unit of the gastroenterology department of a large, tertiary-care referral academic hospital. Participants were 214 consecutive patients, enrolled over a 5-month period, including 116 with Crohn's disease and 53 with ulcerative colitis, as well as 45 with other gastrointestinal diseases and 51 healthy controls. Anti-Saccharomyces cerevisiae antibodies showed the best performance (54% sensitivity and 88%-95% specificity for Crohn's disease). Among patients with negative anti-Saccharomyces antibodies, 19 (34%) had high titers of at least another tested antiglycan antibody. Anti-Saccharomyces and anti-laminaribioside antibodies were associated with disease involving the small bowel and with penetrating or stricturing phenotype. Anti-laminaribioside was significantly higher in patients with a familial history of inflammatory bowel disease. The new proposed serological markers are significantly associated with Crohn's disease, with low sensitivity but good specificity. About one third of anti-Saccharomyces-negative patients may be positive for at least 1 of those markers. Antiglycan antibodies appear to be associated with characteristic localization and phenotype of the disease.


A 52-week, randomized, open-label, parallel-group comparison of the tolerability and effects of pitavastatin and atorvastatin on high-density lipoprotein cholesterol levels and glucose metabolism in Japanese patients with elevated levels of low-density lipoprotein cholesterol and glucose intolerance.
Statin therapy has been found to produce substantial reductions in low-density lipoprotein cholesterol (LDL-C) levels, resulting in a reduced risk for cardiovascular events. Recently, research interest has focused on modification of high-density lipoprotein cholesterol (HDL-C) levels for the potential prevention of cardiovascular events. The effects of pitavastatin and atorvastatin on HDL-C have not been directly compared. This study compared the effects of pitavastatin and atorvastatin on HDL-C and other lipids and glucose metabolism in Japanese patients with elevated LDL-C levels and glucose intolerance. The tolerability of the 2 treatments was also compared. This was a multicenter, open-label, parallel-group trial. Patients with LDL-C levels>or=140 mg/dL and glucose intolerance (defined according to Japanese criteria for borderline diabetes and World Health Organization criteria for impaired fasting glucose and impaired glucose tolerance) were randomly assigned to receive either pitavastatin 2 mg/d or atorvastatin 10 mg/d for 52 weeks. Levels of serum lipids and lipoproteins and measures of glucose metabolism (fasting insulin, fasting glucose, glycosylated hemoglobin, and homeostasis model assessment for insulin resistance) were obtained at baseline and at 8, 26, and 52 weeks of treatment. The effect of study drug on glucose metabolism was evaluated as a tolerability outcome. Tolerability was further assessed based on adverse events, either spontaneously reported or elicited by questioning; physical examination findings; and clinical laboratory test results. Study physicians rated the relationship of adverse events to study medication as unrelated, suspected, or probable. Two hundred seven patients were enrolled in the study, and efficacy was evaluated in 173 patients (88 pitavastatin, 85 atorvastatin). Thirty-four patients were excluded for reasons including failure to start medication or lack of >or=6 months of follow-up. Women accounted for 62% (108/173) of the evaluable population, which had a mean age of 63.3 years and a mean weight of 63.0 kg; 89% (154/173) had diabetes mellitus. The percent change in HDL-C levels was significantly greater in the pitavastatin group compared with the atorvastatin group (8.2 vs 2.9, respectively; P=0.031), as was the percent change in apolipoprotein (Apo) A-I (5.1 vs 0.6; P=0.019). The percent change in LDL-C levels was significantly lower with atorvastatin compared with pitavastatin (-40.1 vs -33.0, respectively; P=0.002), as were the percent changes in non-HDL-C (-37.4 vs -31.1; P=0.004), Apo B (-35.1 vs -28.2; P<0.001), and Apo E (-28.1 vs -17.8; P<0.001). The significant results for these parameters were unchanged when all 189 subjects who received>or=1 dose of study medication were included in the analysis, using last-value-carried-forward methodology. There were no significant differences between treatments with respect to the measures of glucose metabolism. Both statins appeared to be well tolerated. Adverse events occurred in 9% (9/96) of the pitavastatin group and 14% (13/93) of the atorvastatin group (P=NS). Two patients in the pitavastatin group and none in the atorvastatin group had an alanine aminotransferase value>3 times the upper limit of normal (P=NS). In these patients with elevated LDL-C levels and glucose intolerance, 52 weeks of treatment with pitavastatin 2 mg/d was associated with significantly greater increases in HDL-C and Apo A-I levels than atorvastatin 10 mg/d. Both treatments were well tolerated.


Off-loading neuropathic wounds associated with diabetes using an ankle-foot orthosis.
Patients with chronic diabetes have a broad spectrum of associated peripheral neurologic deficits that culminate in an increased susceptibility to ulcer formation. The authors focus on the use of the ankle-foot orthosis as both a treatment and a definitive solution for achieving ulcer closure and for minimizing the chance of ulcer recurrence in the ambulatory patient. An analysis of the pathologic forces encountered, and the solution achieved with the ankle-foot orthosis is presented. In addition, the results from a clinical pilot study in subjects with recalcitrant ulcers secondary to Charcot's neuroarthropathy are presented.


Rapid development of Th2 activity during T cell priming.
The paradigm of T helper-1 (Th-1) and Th-2 cells developing from non-committed naive precursors is firmly established. Th1 cells are characterized by IFNgamma production and, in mice, the selective switching to IgG2a. Conversely IL-4 production and selective switching to IgG1 and IgE characterize Th2 cells. Analysis of Th2 induction in vitro indicates that this polarization develops gradually in T cells activated by anti-CD3 in the presence of IL-4; conversely anti-CD3 and IFNgamma induce Th1 cells. In this report, we explore evidence that indicates that the T helper cell polarization in vivo cannot solely be explained by the cytokine environment. This is provided by studying the early acquisition of Th1 and Th2 activities during responses to a mixture of Th1 and Th2-inducing antigens. It is shown that these divergent forms of T cell help can rapidly develop in cells within a single lymph node. It is argued that early polarization to show Th-1 or Th-2 behavior can be induced by signals delivered during cognate interaction between virgin T cells and dendritic cells, in the absence of type 1 or type 2 cytokines. This contrasts with the critical role of the cytokines in reinforcing the Th-phenotype and selectively expanding T helper clones.


[Study of the mechanism of adsorption and activation of enzymes of the contact phase of blood coagulation].
Desorption of enzymes, participating in contact phase of blood coagulation, from silica surface was studied under effect of increased ion strength, in presence of urea, EDTA and surface-active substances. All these agents varied in their capacity to decrease the enzymes activity and to cause the desorption of contact factors. Ionic and hydrogen bonds were shown to carry out the leading role in adsorption of factor XII and in the initiation of the contact phase of blood coagulation.


Metabolite profiling of wheat flag leaf and grains during grain filling phase as affected by sulfur fertilisation.
Increasing prices for wheat products and fertilisers call for an adjusted agricultural management to maintain yield and to improve product quality. With the increased use of sulfur-free fertilisers in modern cropping systems and the decrease of atmospheric sulfur emissions by industry, sulfur has become a major limiting factor for crop production. The presented data showed that by using GC-MS it was possible to quantitatively detect a set of 72 different metabolites including amino acids, organic acids, sugars, sugar phosphates, and sugar alcohols, phenolic compounds and nucleotides from wheat grains and flag leaves of a pot experiment. A principal component analysis (PCA) revealed a clear separation of flag leaves and grains and a clear separation of non-fertilised and fertilised flag leaves. It could further be shown by PCA, that the low level sulfur fertilisation is also separated from the higher fertilised grains. A considerable influence of the sulfur fertilisation not only on sulfur rich amino acids but also on the sugar metabolism was detected. With increasing sulfur fertilisation six sugars and sugar derivates in the grain such as glucose-6P, galactose, trehalose, cellobiose, melibiose, fumarate, glycerate and the nucleotide uracil were enhanced. Therefore, it was concluded that photosynthesis was limited in developing plants suffering from sulfur deficiency. Late sulfur fertilisation is a procedure that can help to prevent sulfur deficiency. A latent sulfur deficiency at ear emergence can be compensated by late sulfur fertilisation, as wheat plants can replenish sulfate deficits within a short time.


Optimization and Analysis of a U-Shaped Linear Piezoelectric Ultrasonic Motor Using Longitudinal Transducers.
A novel U-shaped piezoelectric ultrasonic motor that mainly focused on miniaturization and high power density was proposed, fabricated, and tested in this work. The longitudinal vibrations of the transducers were excited to form the elliptical movements on the driving feet. Finite element method (FEM) was used for design and analysis. The resonance frequencies of the selected vibration modes were tuned to be very close to each other with modal analysis and the movement trajectories of the driving feet were gained with transient simulation. The vibration modes and the mechanical output abilities were tested to evaluate the proposed motor further by a prototype. The maximum output speed was tested to be 416 mm/s, the maximum thrust force was 21 N, and the maximum output power was 5.453 W under frequency of 29.52 kHz and voltage of 100 Vrms. The maximum output power density of the prototype reached 7.59 W/kg, which was even greater than a previous similar motor under the exciting voltage of 200 Vrms. The proposed motor showed great potential for linear driving of large thrust force and high power density.


Altered renal function in broilers during aflatoxicosis.
Experiments were conducted to determine the effects of aflatoxicosis on acid-base balance, urine flow rate (V), glomerular filtration rate (GFR), clearance of para-aminohippuric acid (CPAH), plasma osmolality, and the renal handling of Na, K, Ca, and P. Three-week-old broilers were gavaged with aflatoxin at a dose of 2 mg/kg of BW per day for 10 consecutive days. Control birds received an equal volume of corn oil, the aflatoxin carrier vehicle. On the eleventh day, the birds were anesthetized and prepared for renal function analysis. A solution containing inulin, para-aminohippuric acid, and mannitol was infused at a low infusion rate (.1 mL/kg of BW per min) and a high infusion rate (.4 mL/kg of BW per min) to determine if aflatoxin affects the renal response to an acute volume load. Aflatoxicosis decreased the fractional excretion of phosphorous (FEP) and plasma Ca concentration but did not significantly alter any other renal function or acid-base variables. The decrease in FEP and plasma Ca may be a direct result of renal tubular damage, decreased Ca absorption from the gut, or a result of altered circulating levels of parathyroid hormone (PTH), and possibly decreased renal sensitivity to PTH.


GRP78/BiP determines senescence evasion cell fate after cisplatin-based chemotherapy.
Cisplatin (CDDP) induces senescence characterized by senescence-associated secretory phenotypes (SASP) and the unfolded protein response (UPR). In this study, we investigated the proteins related to the UPR during the senescence cell fate. Strikingly, we found that one of the critical ER-resident proteins, GRP78/BiP, was significantly altered. Here we show that GRP78 levels differentially expressed depending on non-small lung cancer subtypes. GRP78 indeed regulates the evasion of senescence in adenocarcinoma A549 cells, in which the increased GRP78 levels enable them to re-proliferate after CDDP removal. Conversely, GRP78 is downregulated in the senescence H460 cells, making them lacking senescence evasion capability. We observed that the translational regulation critically contributed to the GRP78 protein levels in CDDP-induces senescence. Furthermore, the increased GRP78 level during senescence confers resistance to senolytic drug, Bortezomib, as observed by a twofold increase in IC50 in A549 senescence cells compared to the wild-type. This observation is also consistent in the cells that have undergone genetic manipulation by transfection with pcDNA3.1(+)-GRP78/BiP plasmids and pSpCas9(BB)-2A-Puro containing guide RNA sequence targeting GRP78 exon 3 to induce the overexpression and downregulation of GRP78 in H460 cells, respectively. Our findings reveal a unique role of GRP78 on the senescence evasion cell fate and senolytic drug resistance after cisplatin-based chemotherapy.


Prevalence and correlates of anemia among adolescents in Nepal: Findings from a nationally representative cross-sectional survey.
Anemia is regarded as major public health problem among adolescents in Low and Middle-Income Countries (LMICs) but there is limited primary data in many countries, including Nepal. This study investigated the prevalence and correlates of anemia in a nationally representative sample of adolescents within the 2014 National Adolescent Nutrition Survey in Nepal. A total of 3780 adolescents aged 10 to 19 years were selected from a cross-sectional survey through multi-stage cluster sampling. Structured interviews, anthropometric measurements and hemoglobin assessments of capillary blood were obtained. Bivariate and multivariable analyses were undertaken to compute the Adjusted Odds Ratio (aOR) for socio-demographic, behavioral and cluster characteristics. The overall prevalence of anemia was 31% (95%CI: 28.2, 33.5), 38% (95%CI: 34.0, 41.8) in female and 24% (95%CI: 20.6, 27.1) in male. The likelihood of anemia was significantly higher among older adolescents (aOR 1.75, 95%CI: 1.44, 2.13), females (aOR 2.02; 95%CI: 1.57, 2.60), among those who walk barefoot (aOR 1.78, 95%CI: 1.08, 2.94), and those residing in the Terai (aOR 1.80, 95%CI: 1.18, 2.77). Food consumption from more than four food groups (aOR 0.71, 95%CI: 0.57, 0.88) was protective against anemia. In conclusion, anemia is common in Nepali adolescents. Efforts to improve the nutritional status of this high-risk age group require nutrition that focus on eating habits, sanitation, iron supplementation and the treatment of hookworm infection.


Calmodulin differentially modulates Smad1 and Smad2 signaling.
The members of the Smad protein family are intracellular mediators of transforming growth factor beta (TGF-beta) signaling. Smad1 transduces bone morphogenetic protein signals, inducing formation of ventral mesoderm in Xenopus embryos, whereas Smad2 transduces activin/TGF-beta signals, generating dorsal mesoderm. Calmodulin directly binds to many Smads and was shown to down-regulate Smad2 activity in a cell culture system (Zimmerman, C. M., Kariapper, M. S. T., and Mathews, L. S. (1997) J. Biol. Chem. 273, 677-680). Here, we extend those data and demonstrate that calmodulin alters Smad signaling in living embryos, increasing Smad1 activity while inhibiting Smad2 function. To characterize this regulation, we undertook a structure-function analysis and found that calmodulin binds to two distinct and conserved regions in both Smad1 and Smad2. Receptor tyrosine kinase signaling also modifies Smad activity (Kretzschmar, M., Doody, J., and Massagué, J. (1997) Nature 389, 618-622; Kretzschmar, M., Doody, J., Timokhina, I., and Massagué, J. (1999) Genes Dev. 13, 804-816; de Caestecker, M. P., Parks, W. T., Frank, C. J., Castagnino, P., Bottaro, D. P., Roberts, A. B., and Lechleider, R. J. (1998) Genes Dev. 12, 1587-1592). We show that calmodulin binding to Smads inhibits subsequent Erk2-dependent phosphorylation of Smads and vice versa. These observations suggest the presence of a cross-talk between three major signaling cascades as follows: Ca(2+)/calmodulin, receptor tyrosine kinase, and TGF-beta pathways.


MiR-154 inhibits cells proliferation and metastasis in melanoma by targeting AURKA and serves as a novel prognostic indicator.
Increasing evidence suggested that dysregulated miR-154 in several tumor tissues is involved in the clinical progress of cancers patients. The objective of this study was to explore the expression pattern of miR-154 and its potential effects in human melanoma. Microarray data from GEO datasets were analyzed to identify differentially expressed miRNAs. Real Time-Polymerase Chain Reaction (RT-PCR) was performed to determine the expressions of miR-154 in melanoma cell lines and tumor tissues. The associations between miR-154 levels and clinical progress were studied using a series of statistical methods. Cell viability, invasion, migration, and apoptosis were detected by Cell Counting Kit-8 (CCK-8) assays, transwell assay, wound healing assays, and flow cytometry, respectively. TargetScan system was used to identify the target genes of miR-154 and Luciferase activity analysis was carried out to demonstrate the possible target. The expression levels of miR-154 were distinctly lower in tumor samples and melanoma cell lines than in normal controls (p < 0.01). The up-regulation of miR-154 in melanoma tissues was associated with advanced tumor stage (p = 0.028), ulceration (p = 0.046), and shorter overall survival (p = 0.0035). Moreover, the multivariate analysis suggested a decreased expression of miR-154 is an independent predictor of overall survival rates in melanoma patients. Functional observation showed that up-regulation of miR-154 suppressed the capability of proliferation, invasion, and migration, promoting apoptosis in melanoma cell lines. Bioinformatics analysis predicted AURKA (aurora kinase A) as a target of miR-154, which was confirmed using the luciferase activity assays. Besides, miR-154 overexpression rescued the suppressive effect of AURKA-mediated melanoma on cell proliferation, colony formation, and metastasis. These results revealed that miR-154 has clinical implications for targeted therapy of melanoma patients and indicated that miR-154 could represent a novel biomarker in predicting the clinical outcome for melanoma.


[Synthesis of (+/-)-edulinine analogues].
The natural product, edulinine (I), possesses analgesic, anaesthetic and anticonvulsant activities on the central nervous system. In this paper we report that a series of edulinine analogues IV, V were synthesized and screened in mice according to methods of Topliss and Austel quantitative drug design. In preliminary pharmacological tests a number of synthesized compounds showed central depressant activities stronger than edulinine.


Chemotherapy for anaplastic thyroid cancer using docetaxel and cisplatin: report of eight cases.
Anaplastic thyroid carcinoma has a dismal prognosis and lacks an established therapeutic strategy. We have recently conducted chemotherapy with docetaxel and cisplatin as part of multimodal treatment for eight patients with anaplastic thyroid carcinoma. Docetaxel (75 mg/m2) and cisplatin (75 mg/m2) were administered on day 1 every 4 weeks for six courses. This chemotherapy was used as induction therapy in one patient, as therapy for distant metastases in five patients and as postoperative adjuvant therapy in two patients. Three patients showed partial responses and three patients showed stable disease. After excluding the two patients receiving the treatment as adjuvant therapy the response rate was 50 %. Grade 3 or 4 leukocytopenia occurred in seven patients (88 %), but these adverse events were tolerable. Chemotherapy with docetaxel and cisplatin may thus be feasible for anaplastic thyroid carcinoma.