Quantitative Modeling and Simulation in PMDA: A Japanese Regulatory Perspective.
In Japan in October 2016, the Pharmaceuticals and Medical Devices Agency (PMDA) began to receive electronic data in new drug applications (NDAs). These electronic data are useful to conduct regulatory assessment of sponsors' submissions and contribute to the PMDA's research. In this article, we summarize the number of submissions of quantitative modeling and simulation (M&S) documents in NDAs in Japan, and we describe our current thinking and activities about quantitative M&S in PMDA.


The oncogenic effects of p53-inducible gene 3 (PIG3) in colon cancer cells.
The p53-inducible gene 3 (PIG3), initially identified as a gene downstream of p53, plays an important role in the apoptotic process triggered by p53-mediated reactive oxygen species (ROS) production. Recently, several studies have suggested that PIG3 may play a role in various types of cancer. However, the functional significance of PIG3 in cancer remains unclear. Here, we found that PIG3 was highly expressed in human colon cancer cell lines compared to normal colonderived fibroblasts. Therefore, we attempted to elucidate the functional role of PIG3 in colon cancer. PIG3 overexpression increases the colony formation, migration and invasion ability of HCT116 colon cancer cells. Conversely, these tumorigenic abilities were significantly decreased in in vitro studies with PIG3 knockdown HCT116 cells. PIG3 knockdown also attenuated the growth of mouse xenograft tumors. These results demonstrate that PIG3 is associated with the tumorigenic potential of cancer cells, both in vitro and in vivo, and could play a key oncogenic role in colon cancer.


Evidence appraisal: a scoping review, conceptual framework, and research agenda.
Critical appraisal of clinical evidence promises to help prevent, detect, and address flaws related to study importance, ethics, validity, applicability, and reporting. These research issues are of growing concern. The purpose of this scoping review is to survey the current literature on evidence appraisal to develop a conceptual framework and an informatics research agenda. We conducted an iterative literature search of Medline for discussion or research on the critical appraisal of clinical evidence. After title and abstract review, 121 articles were included in the analysis. We performed qualitative thematic analysis to describe the evidence appraisal architecture and its issues and opportunities. From this analysis, we derived a conceptual framework and an informatics research agenda. We identified 68 themes in 10 categories. This analysis revealed that the practice of evidence appraisal is quite common but is rarely subjected to documentation, organization, validation, integration, or uptake. This is related to underdeveloped tools, scant incentives, and insufficient acquisition of appraisal data and transformation of the data into usable knowledge. The gaps in acquiring appraisal data, transforming the data into actionable information and knowledge, and ensuring its dissemination and adoption can be addressed with proven informatics approaches. Evidence appraisal faces several challenges, but implementing an informatics research agenda would likely help realize the potential of evidence appraisal for improving the rigor and value of clinical evidence.


Nitric Oxide Regulation of H-NOX Signaling Pathways in Bacteria.
Nitric oxide (NO) is a freely diffusible, radical gas that has now been established as an integral signaling molecule in eukaryotes and bacteria. It has been demonstrated that NO signaling is initiated upon ligation to the heme iron of an H-NOX domain in mammals and in some bacteria. Bacterial H-NOX proteins have been found to interact with enzymes that participate in signaling pathways and regulate bacterial processes such as quorum sensing, biofilm formation, and symbiosis. Here, we review the biochemical characterization of these signaling pathways and, where available, describe how ligation of NO to H-NOX specifically regulates the activity of these pathways and their associated bacterial phenotypes.


Variation in phosphoric acid concentration and treatment time and HEMA diffusion through dentin.
The effects of phosphoric acid concentration and time of dentin treatment on the movement of 2-hydroxyethylmethacrylate (HEMA) from a bonding resin-resin composite combination through dentin were investigated in vitro. Freshly extracted human third molar teeth were divided into seven groups each of 10 teeth. A closed chamber was attached to the CEJ of each tooth to contain 1 ml distilled water. An occlusal cavity of 6 mm diameter and remaining dentin thickness of 1.0-1.5 mm was prepared in each tooth. Dentin was treated with either 10% phosphoric acid for 15, 30 or 60 seconds or with 37% phosphoric acid for 15, 30 or 60 seconds. A control group not treated with acid was also prepared. The cavities were rinsed, dried and then treated with the HEMA-containing Scotchbond Multi-Purpose bonding resin which was light-cured for 10 seconds. The cavities were then restored with Z100 and light-cured for 30 seconds. Water samples were retrieved from the chambers over a time course (4.32, 14.4, 43.2, 144 & 432 minutes; 1, 3 and 10 days) and analyzed by high performance liquid chromatography. HEMA was detected in the pulp chambers of all teeth from 4.32 minutes after resin placement. Mean total (cumulative) release at 10 days for all groups was in the 2-4 mumol range. The highest HEMA diffusion rate was 4.32 minutes after placement and were in the range 6-13 times greater than control in the 10%-15 seconds, 10%-30 seconds, 10%-60 seconds, 37%-15 seconds, and 37%-30 seconds groups. Unexpectedly, by far the lowest early rate at 4.32 minutes was in the 37%-60 seconds group (0.6 times of no-acid control).


[Shutter-glasses haploscope with eye-tracking control for fusion therapy].
Common haploscopes require a manual input of the presented disparity and particularly prepared images; moreover, they can be used predominantly by orthoptists. We developed a device that makes possible a haploscope on the basis of a computer monitor with shutter-glasses. The presented disparity is determined with help of an eye-tracker. A patient looks at a monitor through shutter-glasses, the eyes being recorded by a camera. An eye-tracking software determines the fixation points and the angle of strabismus. Images are presented at the monitor with a disparity that is regulated according to these measurements. This is controlled by means of a software so that the disparity is gradually decreased according to the patients abilities. Diagnostic data are recorded permanently. Any monitor contents, such as videos, TV or computer applications, can be used for fusion training or diagnostic purposes with the new apparatus. Initial investigations of patients with heterophoria or intermittent exophoria indicate a decrease of the subjective angle of strabismus by compensation and an increase of the fusional range. The developed shutter-glass-haploscope with eye-tracking control provides several methods for diagnosis. The appliance provides automated fusion training adapted to the patient that can be done at the working place or at home.


Macrolactonization of Alkynyl Alcohol through Rh(I)/Yb(III) Catalysis.
A catalytic macrolactonization through oxidative cyclization of alkynyl alcohol by synergistic transition-metal and Lewis-acid catalysis was developed. Because the alkynyl alcohol substrates involved in this method are different from the seco acids that are used in conventional macrolactonization methods, the current method provides a strategically distinct entry to macrolactones. In addition to the operational simplicity, this macrolactonization protocol proceeds at relatively high concentration, precluding the need for high dilution or slow addition procedures.


A national survey of asthma knowledge and practices among specialists and primary care physicians.
Asthma is a chronic disorder that causes significant morbidity and mortality and requires ongoing chronic care. Approximately two-thirds of people with asthma are receiving care from a primary care clinician, such as an internist, family practitioner, nurse practitioner, or pediatrician. The other one-third of patients are obtaining treatment and ongoing care from specialists, including allergists or pulmonologists. The outcomes of asthma care are a subject of intense investigation. Many studies focus on pharmacotherapy, allergen control, and asthma education as interventions to reduce the morbidity and costs associated with asthma. Fewer studies have explored the differences in outcomes between asthmatic patients cared for by specialists compared with generalists. Even fewer have explored the practice differences between generalists and specialists that may relate to outcomes of care. With the advent of national asthma guidelines and the high prevalence of asthma seen in primary care settings, it is important to investigate the knowledge, attitudes, and practices of primary care physicians with regard to asthma.


Evaporation of freely suspended single droplets: experimental, theoretical and computational simulations.
Evaporation is ubiquitous in nature. This process influences the climate, the formation of clouds, transpiration in plants, the survival of arctic organisms, the efficiency of car engines, the structure of dried materials and many other phenomena. Recent experiments discovered two novel mechanisms accompanying evaporation: temperature discontinuity at the liquid-vapour interface during evaporation and equilibration of pressures in the whole system during evaporation. None of these effects has been predicted previously by existing theories despite the fact that after 130 years of investigation the theory of evaporation was believed to be mature. These two effects call for reanalysis of existing experimental data and such is the goal of this review. In this article we analyse the experimental and the computational simulation data on the droplet evaporation of several different systems: water into its own vapour, water into the air, diethylene glycol into nitrogen and argon into its own vapour. We show that the temperature discontinuity at the liquid-vapour interface discovered by Fang and Ward (1999 Phys. Rev. E 59 417-28) is a rule rather than an exception. We show in computer simulations for a single-component system (argon) that this discontinuity is due to the constraint of momentum/pressure equilibrium during evaporation. For high vapour pressure the temperature is continuous across the liquid-vapour interface, while for small vapour pressures the temperature is discontinuous. The temperature jump at the interface is inversely proportional to the vapour density close to the interface. We have also found that all analysed data are described by the following equation: da/dt = P(1)/(a + P(2)), where a is the radius of the evaporating droplet, t is time and P(1) and P(2) are two parameters. P(1) = -λΔT/(q(eff)ρ(L)), where λ is the thermal conductivity coefficient in the vapour at the interface, ΔT is the temperature difference between the liquid droplet and the vapour far from the interface, q(eff) is the enthalpy of evaporation per unit mass and ρ(L) is the liquid density. The P(2) parameter is the kinetic correction proportional to the evaporation coefficient. P(2) = 0 only in the absence of temperature discontinuity at the interface. We discuss various models and problems in the determination of the evaporation coefficient and discuss evaporation scenarios in the case of single- and multi-component systems.


Issues and considerations when researching sensitive issues with men: examples from a study of men and sexual health.
This article highlights some ethical considerations and practicalities that arose when conducting research with men in relation to decision making and seeking help for a sexually transmitted infection (STI). The aim of the study was to explore, using a grounded theory approach, the processes involved in making the decision to seek help. The main issues that arose were in relation to seeking permission and gaining approval, obtaining informed consent and susceptibility to harm. The interview itself and the concept of the researcher as a tool of data collection also gave rise to ethical issues. These issues have implications for the conduct of sensitive research with men and highlight the need for more overt reporting of the ethical issues and practical considerations that arise when planning and conducting research.


Characterization of T-regulatory cells, induced by immature dendritic cells, which inhibit enteroantigen-reactive colitis-inducing T-cell responses in vitro and in vivo.
Regulatory T (Treg) cells, derived from co-cultures of unfractionated CD4(+) T cells and immature dendritic cells (DC), suppress enteroantigen-induced proliferation of CD4(+) CD25(-) T cells. The DC-induced Treg cells are a mixture of CD25(+) (10-20%) and CD25(-) (80-90%) T cells. However, all the suppressor activity in vitro and in vivo resides in the CD25(+) T-cell subset. The CD25(+) DC-induced Treg cells can inhibit enteroantigen-induced proliferation in vitro through a transwell membrane, and their function does not appear to depend on previous activation. DC-induced CD25(+) Treg cells display a naive phenotype, expressing high levels of CD45RB and l-selectin (CD62L). In addition, the DC-induced Treg cells mediate a stronger suppressive activity than prototype CD25(+) regulatory T cells. The DC-induced Treg cells, and hereof purified CD25(+) and CD25(-) T-cell fractions, were co-injected into severe combined immunodeficiency (SCID) mice with colitis-inducing CD4(+) CD25(-) T cells. Both unfractionated CD4(+) and purified CD25(+) Treg cells fully protected the recipients against the development of colitis. In contrast, co-transfer of fractionated CD25(-) T cells offered no protection against disease development. Enterobacterial antigen-exposed CD4(+) T cells of the protected mice secreted higher levels of interleukin-10 and lower levels of interferon-gamma than the unprotected mice. The present data demonstrate DC-induced CD4(+) CD25(+) Treg cells, which phenotypically and functionally differ from the generally accepted prototype of CD25(+) Treg cells. These data may initiate new procedures for the expansion of Treg cells for clinical use.


The two faces of TOE-2.
The C. elegans Q lineage provides a unique context for studying how cells divide asymmetrically to generate cells fated to die. The Q cell divides to form the Q.a and Q.p neuroblasts, each of which divides to produce neurons and a cell that dies by apoptosis; however, these neuroblasts employ different mechanisms to divide asymmetrically.(1) We discovered 2 distinct roles for TOE-2, a protein previously shown to be a target of the C. elegans ERK ortholog MPK-1, in promoting apoptosis in each of these neuroblast divisions. In this commentary, we discuss possible molecular mechanisms by which TOE-2 promotes apoptosis. Specifically, we will discuss potential roles for TOE-2 interacting proteins, a possible nuclear function for TOE-2, and a potential link to the Wnt pathway.


Parsonage-Turner Syndrome Following COVID-19 Vaccination: Clinical and Electromyographic Findings in 6 Patients.
Parsonage-Turner syndrome (PTS) following COVID-19 infection or vaccination is rare. The pathophysiology may involve an immune-mediated inflammatory reaction against brachial plexus nerve fibers in a genetically predisposed individual. We describe the clinical and electromyographic features of 6 patients presenting with the clinical picture of PTS following COVID-19 vaccination. All patients were referred for electromyographic studies to evaluate the acute onset of pain in the shoulder girdle/upper limb accompanied by muscle weakness in the distribution of one or more branches of the brachial plexus. Each patient had received the COVID-19 vaccine within a few weeks prior to the onset of symptoms. Patients underwent detailed neurological examinations followed by nerve conduction and EMG studies. The patients developed symptoms after a mean duration of 17 days (5 days-8 weeks) after receiving the COVID-19 vaccine. The initial symptom was pain in the shoulder girdle/upper limb, followed within days by muscle weakness. Physical examinations and EMG studies showed upper trunk brachial plexopathy in 2 patients, lower trunk plexopathy in 1 patient, posterior cord brachial plexopathy in 1 patient, and anterior/posterior interosseous nerve involvement in 2 patients. All patients either improved or attained complete resolution of the arm pain at follow-up. Three (50%) patients did not have any improvement in the arm/hand weakness, while 3 (50%) patients had some recovery in strength. PTS may occur after the COVID-19 vaccine and should be suspected in patients with symptoms and signs suggestive of acute brachial plexopathy. Studies of a larger series may provide insight into predisposing factors.


The Recombinant Protein EphB4-Fc Changes the Ti Particle-Mediated Imbalance of OPG/RANKL via EphrinB2/EphB4 Signaling Pathway and Inhibits the Release of Proinflammatory Factors In Vivo.
Aseptic loosening caused by wear particles is one of the common complications after total hip arthroplasty. We investigated the effect of the recombinant protein ephB4-Fc (erythropoietin-producing human hepatocellular receptor 4) on wear particle-mediated inflammatory response. In vitro, ephrinB2 expression was analyzed using siRNA-NFATc1 (nuclear factor of activated T-cells 1) and siRNA-c-Fos. Additionally, we used Tartrate-resistant acid phosphatase (TRAP) staining, bone pit resorption, Enzyme-linked immunosorbent assay (ELISA), as well as ephrinB2 overexpression and knockdown experiments to verify the effect of ephB4-Fc on osteoclast differentiation and function. In vivo, a mouse skull model was constructed to test whether the ephB4-Fc inhibits osteolysis and inhibits inflammation by micro-CT, H&E staining, immunohistochemistry, and immunofluorescence. The gene expression of ephrinB2 was regulated by c-Fos/NFATc1. Titanium wear particles activated this signaling pathway to the promoted expression of the ephrinB2 gene. However, ephrinB2 protein can be activated by osteoblast membrane receptor ephB4 to inhibit osteoclast differentiation. In in vivo experiments, we found that ephB4 could regulate Ti particle-mediated imbalance of OPG/RANKL, and the most important finding was that ephB4 relieved the release of proinflammatory factors. The ephB4-Fc inhibits wear particle-mediated osteolysis and inflammatory response through the ephrinB2/EphB4 bidirectional signaling pathway, and ephrinB2 ligand is expected to become a new clinical drug therapeutic target.


The effects of nitric oxide synthase blockers on mesenteric blood flow with bile duct ligation.
The aim of this study was to investigate the effects of nitric oxide synthesis inhibition by L-NNA (N-nitro-L-arginine) on mesenteric blood flow in bile duct ligated rats. Forty female Wistar rats were allocated into four groups. Only common bile duct (CBD) was dissected with midline laparotomy (sham operated), CBD is ligated for obstructive jaundice (OJ group), with CBD dissection, 20 mg/kg L-NNA was given i.p. during 14 days (sham + L-NNA group), after CBD ligation, 20 mg/kg L-NNA was given i.p. during 14 days (OJ + L-NNA group). At the end of the 14th day, the rats were re-anaesthesized, phenylephrine was administered within the left jugular vein and, arterial blood pressure monitorization was done via left carotid artery. Blood flow of the mesenteric artery (SMA) was also recorded. Resistance of the SMA was calculated and in addition, pressure responses to phenylephrine were detected. Histopathological examination of the liver and blood analyses were done. The mean blood pressure levels of the rats with OJ were lower than the control group (P>0.05). Blood flow of the SMA and vascular resistance of the rats with OJ have no significant difference with other groups. In CBD ligated animals, mean blood pressure and mesenteric vascular resistance increased by L-NNA but there was no difference in mesenteric blood flow rate and vasoconstrictive response to phenylephrine. On microscopic examination of the liver with OJ, bile duct proliferation and minimal mononuclear reaction were observed which was not affected by L-NNA administration. Hypotension, low systemic vascular resistance, and a reduced sensitivity to vasoconstrictors are features of cirrhosis. In obstructive jaundice impaired vascular responsiveness to vasoconstrictor stimuli like as cirrhosis was not observed and also there was no improvement by L-NNA on histopathologic findings and liver function tests.


Use of oral combination therapy for type 2 diabetes in primary care: Meeting individualized patient goals.
The management of type 2 diabetes mellitus (T2DM) by primary care physicians (PCPs) has become increasingly complex due to limitations on consultation time, an increasing array of drug treatment options, and issues of comorbidities and polypharmacy. Diabetes is a progressive condition and treatment with a single glucose-lowering agent can only address limited pathophysiologic targets and does not provide adequate glycemic control in many cases. Consequently, most patients with T2DM will eventually require treatment with multiple glucose-lowering medications. Oral combination therapy in T2DM may be given as multiple-pills, or as single-pill, fixed-dose combinations (FDCs), the latter of which offer convenience, ease of administration, and a reduction in the medication burden. Therefore, FDCs can potentially improve patients' treatment adherence and optimize achievement and maintenance of glycemic targets. However, cost factors also need to be considered. An understanding of the issues associated with the use of combination therapy in T2DM will help PCPs to guide patient-centered decision making and promote the effective management of T2DM.


Multi-Flexible Body Dynamics Modeling and Experimental Study of the Patient Rehabilitation Transfer Device.
The patient rehabilitation transfer device is a typical personnel transfer equipment, which is mainly composed of outriggers, support arm, lifting arms, hooks, handrails, hydraulic cylinders, and other components. The existing research on the device is mainly focused on the configuration design and transfer mode, and the research on its dynamic characteristics during the transfer process has not been thoroughly discussed. Methodology. Based on the existing research, a portable hydraulic rehabilitation patient transfer device has been developed. Then the multi-rigid body dynamic and finite element flexible body models were established. Next, the dynamic characteristic difference between the two models of the device was studied. As shown in the results, the finite element multi-flexible body model has obvious flexible vibration in the lifting stage, and the amplitude reaches 16 mm in the motion startup stage due to the influence of rigid-flexible coupling. The tip acceleration of the flexible body model was also influenced by the vibration, and the maximum acceleration value reaches 0.06 m/s2. According to the test results, the maximum acceleration of the terminal reaches 0.05 m/s2, which is close to the finite-element multi-flexible model simulation results. The experimentally measured natural frequency of the device is 3.1 Hz, which is also close to 3.2 Hz calculated by the simulation. Because the flexible component in the flexible model is only the lift arm, the natural frequency is slightly larger than the experimental value. According to the stress value of the finite element multi-body model motion process, the maximum stress appears at the moment when the motion reaches the top end, and the instantaneous stress reaches 206 Mpa, which is in line with the allowable stress range of the material design. The data obtained in this study will provide help for the follow-up clinical rehabilitation and intelligent device research.


Controlling the photophysical properties of ionic transition-metal complexes through various counterions.
Ionic transition-metal complexes (iTMCs) with rich excited-state properties and excellent optical properties have attracted considerable attention from both the scientific and industrial community in recent decades. Controlling their photophysical properties is very important for realizing a diverse range of optoelectronic applications. Generally, the optical property modulation of iTMCs is achieved by covalent structural modification of their ligand skeleton. In this feature article, we highlight the full potential of the regulation of the optical properties of iTMCs through changing various counterions, which is demonstrated to be a facile and convenient method. This paper lists different counterions that can be used to tune the emission wavelengths, emission lifetimes, and response behaviors of iTMCs. This paper also summarizes the advances towards multi-level information encryption and high-level anti-counterfeiting applications. Moreover, the recent challenges relating to this topic are examined alongside various future directions. We anticipate that the research provided in this paper will assist in directing future analyses based on the regulation of the photophysical properties of iTMCs in a facile, reproducible, and convenient way.


The neurobiological basis of lateralized cerebral function. A review.
The first part of this review considers the empirical findings regarding the anatomical asymmetries that may be related to the left hemisphere's dominance for language. The review reveals that there are interhemispheric differences in the anatomy of the posterior sylvian region and of portions of the inferior frontal gyrus. In the second section, difficulties in attributing functional significance to these morphological asymmetries are discussed. The third section considers anatomical, behavioural and neurochemical asymmetries in animals and their relation to problems of human cerebral asymmetry.


Chemical Space Mimicry for Drug Discovery.
We describe a new library generation method, Machine-based Identification of Molecules Inside Characterized Space (MIMICS), that generates sets of molecules inspired by a text-based input. MIMICS-generated libraries were found to preserve distributions of properties while simultaneously increasing structural diversity. Newly identified MIMICS-generated compounds were found to be bioactive as inhibitors of specific components of the unfolded protein response (UPR) and the VEGFR2 pathway in cell-based assays, thus confirming the applicability of this methodology toward drug design applications. Wider application of MIMICS could facilitate the efficient utilization of chemical space.


High resolution linkage and linkage disequilibrium analyses of chromosome 1p36 SNPs identify new positional candidate genes for low bone mineral density.
A quantitative trait locus (QTL) for BMD maps to chromosome 1p36. We have analyzed a high density SNP panel from this region for linkage and association to BMD in 39 osteoporosis pedigrees. Our results support the presence of genes controlling BMD on 1p36 and indicate new candidates for further analyses. Low BMD is one of the major risk factors for osteoporosis. Following a genome scan in a sample of Caucasian families recruited through probands with low BMD, a region on 1p36 near marker D1S214 received support as a QTL for BMD from linkage (maximum lod-score = 2.87) and linkage disequilibrium (LD) analysis (p < 0.01). To better characterize the genetic risk factors for low BMD located in this genomic region, we have genotyped the same group of families for 1095 SNPs located across 11 Mb on 1p36. Linkage and LD analyses have been performed using the variance component approach. Multivariate linkage analysis indicated two QTLs for femoral neck BMD, lumbar spine BMD and trochanter BMD simultaneously on 1p36, with maximum lod-scores of 4.37 at 12 cM and 3.59 at 22 cM. LD analysis identified several SNPs potentially associated with BMD, including the RERE gene SNP rs11121179 (p = 0.000005 for lumbar spine BMD). Other candidate genes include G1P2, SSU72 and CCDC27 (each containing 1 SNP with p < 0.001 for at least one BMD trait). This study supports the presence in 1p36 of QTLs affecting BMD at multiple skeletal sites. Replication of our results in other independent cohorts is warranted.


RNA modifications in brain tumorigenesis.
RNA modifications are emerging as critical regulators in cancer biology, thanks to their ability to influence gene expression and the predominant protein isoforms expressed during cell proliferation, migration, and other pro-oncogenic properties. The reversibility and dynamic nature of post-transcriptional RNA modifications allow cells to quickly adapt to microenvironmental changes. Recent literature has revealed that the deregulation of RNA modifications can promote a plethora of developmental diseases, including tumorigenesis. In this review, we will focus on four key post-transcriptional RNA modifications which have been identified as contributors to the pathogenesis of brain tumors: m6A, alternative polyadenylation, alternative splicing and adenosine to inosine modifications. In addition to the role of RNA modifications in brain tumor progression, we will also discuss potential opportunities to target these processes to improve the dismal prognosis for brain tumors.


Integrating Palliative Care into the Management of Heart Failure with Reduced Ejection Fraction: A Practice Pearl.
Heart failure with reduced ejection fraction is a progressive, undulating syndrome with an unpredictable illness course featuring intermittent symptom exacerbations and periods of stability. The progressive, variable trajectory of the illness burdens patients with myriad threats to physical, emotional, and spiritual functioning, quality of life and complex treatment decisions. Integrating palliative care is a recommended best practice for heart failure management; however, confusion persists about what palliative care comprises in the context of heart failure.


Deep forest model for diagnosing COVID-19 from routine blood tests.
The Coronavirus Disease 2019 (COVID-19) global pandemic has threatened the lives of people worldwide and posed considerable challenges. Early and accurate screening of infected people is vital for combating the disease. To help with the limited quantity of swab tests, we propose a machine learning prediction model to accurately diagnose COVID-19 from clinical and/or routine laboratory data. The model exploits a new ensemble-based method called the deep forest (DF), where multiple classifiers in multiple layers are used to encourage diversity and improve performance. The cascade level employs the layer-by-layer processing and is constructed from three different classifiers: extra trees, XGBoost, and LightGBM. The prediction model was trained and evaluated on two publicly available datasets. Experimental results show that the proposed DF model has an accuracy of 99.5%, sensitivity of 95.28%, and specificity of 99.96%. These performance metrics are comparable to other well-established machine learning techniques, and hence DF model can serve as a fast screening tool for COVID-19 patients at places where testing is scarce.


Immunopathogenesis of allergic disorders: current concepts.
Allergic disorders are a group of immune-mediated disorders that are associated with considerable morbidity and ill health. There has been significant rise in the prevalence of allergy in the last few years. This has heightened interest in uncovering the novel mechanisms involved in etiopathogenesis of allergic disorders. Understanding the pathways underlying allergy will help in developing effective modalities for its prevention and treatment. This review focuses primarily on common IgE-mediated allergic conditions and recent developments in their immunopathogenesis, especially those involving respiratory mucosa.


Women with septic abortion: who, how and why? A prospective study from tertiary care hospital in India.
A prospective descriptive interview based hospital study was carried on 47 women admitted with septic abortion to evaluate their psychosocial, demographic and clinical profile These women were predominantly parous (75%), hindus (60%), between 20-30 years of age (60%) and mostly married (91.4%) house wives (63.8%). More than 90% already had one or more male child. The contraceptive use was dismally low (23.4%). Their knowledge about legalisation, place and persons authorized to conduct abortions was very less, however large majority (87%) underwent abortions within 3 months of pregnancy. Large family, poverty and spacing were the main reasons cited for abortions. Abdominal pain, fever, genital bleeding, diarhoea and abdominal distension were presenting clinical features in order of frequency. Advanced sepsis and associated medical and surgical complications were present in more than half the patients and 6% succumbed to these problems. The current experience was an eye opener for most of them and changed their future attitude. Hence education, economic prosperity, easy access to reproductive health facilities and institutional management of sepsis is the key to make abortions safe.


Static QCD potential at three-loop order.
We compute the purely gluonic contribution to the static QCD potential at three-loop order. This completes computation of the static potential at this order.


References in the history of otology: the importance of being earnest.
Historical vignettes are often included in scientific otological articles. However, they may contain inaccuracies and errors when authors use secondary or compiled texts rather than primary source material. Examples of errors arising from the use of secondary sources are obtained from compiled works in comparison with original publications, with a particular focus on Hildanus' speculum. Different types of inaccuracies may be found in historical vignettes. In Hildanus' works, two different specula are depicted: one was described by himself during his lifetime, and the other after his death, probably by his publisher. Only the posthumous description of Hildanus' speculum has been retained historically, but it is uncertain that it was really developed by Hildanus himself. Other such inaccuracies exist because of confusion with the original terminology of referring to an instrument, such as the otoscopes of Toynbee and Brunton, inaccuracies in dates, misinterpretation of the original text, such as the attribution to Charcot as the first to propose the intracranial division of the eighth nerve in Menière's disease, and problems of translation (e.g., the different translations of the book titled Diseases of the Ear by Wilhelm Kramer). These different examples of inaccuracies show the common problems encountered in writing a historical report. Authors must be aware of such errors and work almost exclusively with original references. Secondary and compiled references should be reserved for a background of discussion and not as a source of direct support.


Bed rest reduces metabolic protein content and abolishes exercise-induced mRNA responses in human skeletal muscle.
The aim was to test the hypothesis that 7 days of bed rest reduces mitochondrial number and expression and activity of oxidative proteins in human skeletal muscle but that exercise-induced intracellular signaling as well as mRNA and microRNA (miR) responses are maintained after bed rest. Twelve young, healthy male subjects completed 7 days of bed rest with vastus lateralis muscle biopsies taken before and after bed rest. In addition, muscle biopsies were obtained from six of the subjects prior to, immediately after, and 3 h after 45 min of one-legged knee extensor exercise performed before and after bed rest. Maximal oxygen uptake decreased by 4%, and exercise endurance decreased nonsignificantly, by 11%, by bed rest. Bed rest reduced skeletal muscle mitochondrial DNA/nuclear DNA content 15%, hexokinase II and sirtuin 1 protein content ∼45%, 3-hydroxyacyl-CoA dehydrogenase and citrate synthase activity ∼8%, and miR-1 and miR-133a content ∼10%. However, cytochrome c and vascular endothelial growth factor (VEGF) protein content as well as capillarization did not change significantly with bed rest. Acute exercise increased AMP-activated protein kinase phosphorylation, peroxisome proliferator activated receptor-γ coactivator-1α, and VEGF mRNA content in skeletal muscle before bed rest, but the responses were abolished after bed rest. The present findings indicate that only 7 days of physical inactivity reduces skeletal muscle metabolic capacity as well as abolishes exercise-induced adaptive gene responses, likely reflecting an interference with the ability of skeletal muscle to adapt to exercise.


[Newborn Screening Program for congenital hypothyroidism of the State of Santa Catarina, Brazil: etiological investigation in the first visit].
To evaluate the etiology of primary congenital hypothyroidism (PCH) identified in the Newborn Screening Program from the state of Santa Catarina, Brazil, from July 2007 to June 2009 in the first visit. A prospective study was performed in 45 patients with PCH. For the etiological diagnosis, history, physical examination, and additional tests (TSH, free thyroxine, thyroglobulin, bone age assessment, thyroid ultrasound) were carried out in the first visit. The etiology was established in the first visit in 53.3% of cases. Thyroid dysgenesis represented 51.11% of the cases, from which 20% showed hypoplastic thyroid, 13.3% showed athyreosis, and 17.7% showed ectopic glands; 2.2% were diagnosed with dyshormonogenesis. Umbilical hernia was the most prevalent sign (48.89%) and 20% had no clinical manifestations. Patients with dysgenesis showed significant differences (p < 0.05) in terms of cesarean section delivery, delayed bone age, and very high serum TSH. The diagnostic approach used at first visit for PCH patients may determine the etiology in 53.3% of cases. Half of patients had thyroid dysgenesis.


Radiating pain during epidural needle insertion and catheter placement cannot be associated with postoperative persistent paresthesia: a retrospective review.
It has been suggested that radiating pain during spinal or epidural needle insertion and catheter placement can be an indicator of needle-related nerve injury. In this study, using a historical cohort, we investigated what factors could be associated with postoperative persistent paresthesia. In addition, we focused on radiating pain during epidural needle insertion and catheterization. This was a retrospective review of an institutional registry containing 21,606 anesthesia cases. We conducted multivariate logistic analysis in 2736 patients, who underwent epidural anesthesia, using the incidence of postoperative persistent paresthesia as a dependent variable and other covariates, including items of the anesthesia registry and the postoperative questionnaire, as independent variables in order to investigate the factors that were significantly associated with the risk of persistent paresthesia. One hundred and seventy-six patients (6.44%) were found to have persistent paresthesia. Multivariate analysis revealed that surgical site at the extremities (odds ratio (OR), 12.5; 95% confidence interval (CI), 2.77-56.4; the reference was set at abdominal surgery), duration of general anesthesia (per 10 min) (OR, 1.02; 95% CI, 1.01-1.03), postoperative headache (OR, 1.78; 95% CI, 1.04-2.95), and days taken to visit the consultation clinic (OR, 1.03; 95% CI, 1.01-1.06) were independently associated with persistent paresthesia. Radiating pain was not significantly associated with persistent paresthesia (OR, 1.56; 95% CI, 0.69-3.54). Radiating pain during epidural procedure was not statistically significantly associated with persistent paresthesia, which may imply that this radiating pain worked as a warning of nerve injury.


Targeting Adhesive Tumor Adventitia via Injectable Electrospun Short Fibers in Perfusion of Intraperitoneal Sporadic Tumors.
Intraperitoneal sporadic tumor is a common and complicated syndrome in cancers, causing a high rate of death, and people find that intraperitoneal chemotherapy (IPC) can treat intraperitoneal sporadic tumors better than intravenous chemotherapy and surgery. However, the effectiveness and side effects of IPC are controversial, and the operation process of IPC is complicated. Herein, the injectable paclitaxel-loaded (PTX-loaded) electrospun short fibers are constructed through a series process of electrospinning, homogenizing, crosslinking, and subsequent polydopamine coating and folate acid (FA) modification. The evenly dispersed short fibers exhibited effective tumor cell killing and good injectable ability, which is convenient to use and greatly improved the complex operation procedure. Mussel-like protein poly-dopamine coating and FA modification endowed short fibers with the ability of targeted adhesion to tumors, and therefore the short fibers further acted as a kind of micro membrane that could release drugs to tumors at close range, maintaining local high drug concentration and prevent paclitaxel killing normal tissues. Thus, the target-adhesive injectable electrospun short fibers are expected to be the potential candidate for cancer treatment, especially the intraperitoneal sporadic tumors, which are hard to treat by surgery or intravenous chemotherapy.


Pulmonary vascular reactivity of spontaneously hypertensive rats is exacerbated in response to the central administration of exogenous nitric oxide.
1. Centrally, nitric oxide (NO) is a sympathoinhibitory substance. Spontaneously hypertensive rats (SHR) have an impaired central nitroxidergic system and, consequently, NO-mediated decrease in sympathetic activity is exacerbated in SHR compared with Wistar-Kyoto (WKY) rats. We have demonstrated previously that acute hypoxic pulmonary vasoconstriction (HPV) is enhanced by central NO administration. Therefore, in the present study, we hypothesized that accentuation of the HPV by NO would be exacerbated in SHR compared with WKY rats. 2. Mean pulmonary arterial pressure, systemic mean arterial blood pressure, cardiac output and heart rate were measured in pentobarbitone-anaesthetized, artificially ventilated, male SHR and WKY rats. The brief, transient response to a bolus intracerebroventricular (i.c.v.) dose of N(G)-nitro-L-arginine methyl ester (L-NAME; 150 microg in 10 microL) was recorded in all rats. Upon recovery, rats were exposed to acute hypoxia (10% O(2) for 4 min) before and after the i.c.v. administration of the NO donor 3-[4-morpholinyl]-sydnonimine-hydrochloride (SIN-1; 100 microg in 10 microL). 3. In WKY rats, central inhibition of NO synthesis by L-NAME caused a mild increase in tonic pulmonary vascular tone and induced a large systemic pressor response. These responses were not observed in SHR. In contrast, SIN-1 failed to alter tonic pulmonary vascular tone, although it enhanced the HPV in WKY rats and, significantly more so, in SHR. 4. These results confirm that accentuation of the HPV by NO is exacerbated in SHR compared with WKY rats. The mechanism(s) by which the HPV is accentuated by central NO remains to be fully elucidated, but is likely to be associated with the sympathoinhibitory effects of NO and, if so, supports the idea that the nitroxidergic system of the SHR is impaired. Further electrophysiological studies are essential to confirm these assumptions.


Active Multitask Learning With Trace Norm Regularization Based on Excess Risk.
This paper addresses the problem of active learning on multiple tasks, where labeled data are expensive to obtain for each individual task but the learning problems share some commonalities across multiple related tasks. To leverage the benefits of jointly learning from multiple related tasks and making active queries, we propose a novel active multitask learning approach based on trace norm regularized least squares. The basic idea is to induce an optimal classifier which has the lowest risk and at the same time which is closest to the true hypothesis. Toward this aim, we devise a new active selection criterion that takes into account not only the risk but also the excess risk, which measures the distance to the true hypothesis. Based on this criterion, our proposed algorithm actively selects the instance to query for its label based on the combination of the two risks. Experiments on both synthetic and real-world datasets show that our proposed algorithm provides superior performance as compared to other state-of-the-art active learning methods.


Posture-cognition interaction during quiet standing in patients with knee osteoarthritis.
The aim of this study was to determine the effects of cognitive task (silent backward counting) on postural control in patients with knee osteoarthritis (OA) (n = 25) as compared with asymptomatic controls (n = 25). Static postural control during quiet standing was assessed under different conditions including: (1) rigid surface-open eyes; (2) rigid surface-closed eyes; (3) foam surface-open eyes; and (4) foam surface-closed eyes. Dependent variables were center of pressure (COP) parameters and cognitive score. The results of this study showed that for most COP parameters, the patients with knee OA had greater postural sway than asymptomatic control subjects. Moreover, while cognitive loading affects postural sway in dual-task compared to single-task conditions, the pattern of change was not different between the two groups. Cognitive loading caused decreased postural sway in both groups. Future studies should select patients with more disability, choose more attention demanding cognitive tasks, and expose the subjects to dynamic balance situations to further explore the effect of cognitive loading on postural performance in this specific patient population.


Pre-transplantation thymic function is associated with the risk of acute graft versus host disease and cytomegalovirus viremia after allogeneic hematopoietic stem cell transplantation.
To analyze the kinetics of T-cell subsets and thymic function reconstitution after allogeneic hematopoietic stem cell transplantation (AHSCT); to determine whether sjTREC (signal joint TCR rearrangement excision circle) and CD31-positive recent thymic emigrant (CD31 + RTE) are correlated with acute graft versus host disease (aGVHD) or CMV (cytomegalovirus) viremia after AHSCT. Forty-nine patients who underwent AHSCT in our institution were prospectively enrolled. Periphery blood samples were collected before conditioning and at 1, 2, 3 months after AHSCT. T-cell subsets were analyzed with flow cytometry. Genomic DNA was purified from peripheral blood mononuclear cells (PBMCs), and sjTREC was quantified by real-time PCR. Impact of sjTREC and CD31 + RTE on aGVHD and CMV viremia was evaluated by univariate and multivariate Cox regression analyses. The analyzed T-cell subsets and sjTREC of patients before AHSCT were all significantly lower than those of healthy donors (p < 0.05). sjTREC and CD31 + RTE were remarkably decreased in 3 months after AHSCT (p < 0.05). Patients with lower pre-transplantation sjTREC and CD31 + RTE level had higher incidence of CMV viremia after AHSCT (p < 0.05). sjTREC/106 PBMCs was negatively correlated with aGVHD (p = 0.024). Thymic function was impaired before transplantation, and was consistently decreased in 3 months after AHSCT. Patients who had lower pre-transplantation sjTREC level were at high risk of aGVHD and CMV viremia after AHSCT, low pre-transplantation CD31 + RTE was correlated with CMV viremia after AHSCT.


Post-treatment occlusal changes in Class II division 2 subjects treated with the Herbst appliance.
The aim of this retrospective study was to analyse and compare the post-treatment occlusal changes of Class II division 2 treatment with the Herbst appliance in early adolescent, late adolescent, and adult subjects. The subjects were 37 Class II division 2 patients (19 females and 18 males) treated at the Orthodontic Department, University of Giessen, Germany. All were in the late mixed or permanent dentition and exhibited a Class II molar relationship > or =0.5 cusp width (CW) bilaterally or > or =1.0 CW unilaterally, an overbite (OB) >3.0 mm, and two upper central incisors retroclined. The subjects were divided into three skeletal maturity groups based on evaluation of hand wrist radiographs: early adolescent (n = 10, stages MP3-E to MP3-FG at start of treatment, age range: 11.3-13.2 years), late adolescent (n = 14, stages MP3-G to MP3-I at start of treatment, age range: 14.1-16.4 years), and adult (n = 13, stages R-I to R-J at the start of treatment, age range: 16.3-25.6 years). Study casts from before treatment (T1), after Herbst-Tip-Edge-Multibracket appliance treatment (T2), and after an average retention time of 27 months (T3) were analysed. Statistical analysis was undertaken using t-tests for paired and unpaired samples. For the whole sample, the molar relationship at T3 was stable in 82.4 per cent, the canine relationship in 82.9 per cent, and OB in 75.7 per cent of the cases. In the different skeletal maturity groups, the stability of the molars, canines, and overbite was as follows: early adolescents: 95.0, 100.0, and 70.0 per cent, respectively; late adolescents: 92.9, 74.1, and 85.7 per cent, respectively; and adults 61.5, 80.8, 69.2 per cent, respectively. Occlusal correction of Class II division 2 malocclusions with Herbst treatment was relatively stable 2 years post-treatment. The outcome of treatment of adolescents was more stable than that of adults.


Quantification of Adeno-Associated Virus with Safe Nucleic Acid Dyes.
Adeno-associated virus (AAV) is the most commonly used viral vector for both biological and gene therapeutic applications. Although many methods have been developed to measure quantity attributes of AAV, they are often technically challenging and time-consuming. Here, we report a method to titer AAV with GelGreen® dye, a safe green fluorescence nucleic acid dye recently engineered by Biotium company (Fremont, CA). This method, hereinafter referred to as GelGreen method, provides a fast (∼30 min) and reliable strategy for AAV titration. To validate GelGreen method, we measured genome titer of an AAV reference material AAV8RSM and compared our titration results with those determined by Reference Material Working Group (ARMWG). We showed that GelGreen results and capsid enzyme-linked immunosorbent assay results are comparable with each other. We also showed that GelRed® dye, a red fluorescence dye from Biotium, can be used to directly "visualize" AAV genome titer on a conventional gel imager, presenting an especially direct approach to estimate viral quantity. Finally, we showed that GelGreen and GelRed dyes can also be used to quantify self-complementary AAV (scAAV) and crudely purified AAV samples. In summary, we described a technique to titer AAV by using new generation of safe DNA dyes. This technique is simple, safe, reliable, and cost efficient. It has potential to be broadly applied for quantifying and normalizing AAV viral vectors.


Effects of ascorbic acid and sodium selenite on growth and redifferentiation in human hepatoma cells and its mechanisms.
After being treated with ascorbic acid (AA) 3 mM + sodium selenite (SS) 1.5 microM, the growth rate and mitotic index of human hepatoma cells BEL-7402 decreased remarkably. The indexes related to cell malignancy were improved, such as cell surface charge obviously decreased, the electrophoresis rate fell from 1.76 microns.s-1.V-1.cm-1 to 0.93, the average of alpha-fetoprotein (alpha-FP) content decreased from 341 micrograms.g-1 protein to 92, and gamma-glutamyl-transpeptidase (gamma-GT) activity from 0.76 U.g-1 protein to 0.19. The indexes related to cell differentiation were affected favourably, such as the level of tyrosine-alpha-ketoglutarate transaminase (TAT) activity increased from 14.2 mumol.g-1 protein to 49.0, and the colonogenic potential decreased 95.3%. These results indicated that hepatoma cells had been successfully induced to redifferentiation by AA + SS. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GPX) were significantly higher, while the activity of catalase (CAT) was slower in the treated group than in the control group. The malondialdehyde (MDA) content decreased slightly, reduced glutathione (GSH) decreased sharply, and H2O2 content increased dramatically. In conclusion, these results indicate that the combination of ascorbic acid and sodium selenite may induce the redifferentiation of hepatoma cells and inhibit cell growth by virtue of enhancing the activities of antioxidative enzymes and reducing the formation of H2O2, and altering the cell redox status. The combination of ascorbic acid and sodium selenite may be a potent anticancer treatment option for human hepatoma cells.


Oviposition of Culex tarsalis (Diptera: Culicidae) Differs on Water Conditioned by Potential Fish and Insect Predators.
The response of egg-laying Culex tarsalis Coquillett (Diptera: Culicidae) to water conditioned by three fish species used for mosquito control and three predatory aquatic insect species was examined in laboratory binary choice experiments. Oviposition by Cx. tarsalis was 72% less on water conditioned with the arroyo chub, Gila orcutti (Eigenmann & Eigenmann) (Cypriniformes: Cyprinidae) relative to control cups containing aged tap water, but no significant difference was found in the numbers of egg rafts laid on water conditioned with the fathead minnow (Pimephales promelas (Rafinesque), Cypriniformes: Cyprinidae) and the control treatment (water aged 24 h). Mosquito oviposition on water conditioned with the predominantly herbivorous/algivorous California Mozambique tilapia hybrid (Oreochromis mossambicus (Peters) × Oreochromis urolepis hornorum L. (Perciformes: Cichlidae)) or predatory insects (nymphs: Sympetrum corruptum (Hagen) (Odonata: Libellulidae); adults: Thermonectus basillaris (Harris) or Cybister fimbriolatus (Say) (Coleoptera: Dytiscidae)) did not differ significantly relative to that onto water aged for 24 h. As compared with water aged 24 h and water conditioned with diving beetles, oviposition by Cx. tarsalis was significantly lower (≥53%) when live predatory diving beetles were present in oviposition cups. Gravid Cx. tarsalis females do not respond equally to putative semiochemicals in water conditioned with the piscine or aquatic insect predators of immature mosquitoes tested here.


Sequential double immunocytochemical staining for in situ identification of an auto-anti-allotype immune response in allotype-suppressed rabbits.
Immunocytochemical staining has been used to detect putative autoimmune B-cells in rabbits undergoing chronic allotype suppression. This condition is seen in heterozygous rabbits exposed perinatally to antibody against the paternal immunoglobulin allotype. Such animals develop lifelong suppression for this allotype and have been used as models for study of antibody-induced disturbance of immune regulation. Normal rabbits deliberately immunized against a heterologous allotype were used to establish the feasibility of identifying cells forming anti-allotypic antibodies in cryostat sections of rabbit lymphoid tissues. Incubation and staining of tissue sections from suppressed rabbits then revealed the presence of autoimmune B-cells, with antibody specificity for the suppressed allotype, in all chronically suppressed adult rabbits tested. Sequential incubation and staining with allotype- and anti-allotype-enzyme conjugates established that such cells were of non-suppressed origin. Auto-anti-allotype antibody-forming cells were not found in normal heterozygotes or in chimeric rabbits. The immunocytochemical techniques described here permitted simultaneous detection of specificity (i.e., anti-allotype) and origin (allotype) of antibody-forming cells involved in an autoimmune response, as well as their anatomical correlation with other B-cells of suppressed or non-suppressed origin. Since the method described can be adapted to detection of alternate cell markers, we believe it to have potential application to the study of other autoimmune phenomena.


Bereavement support standards and bereavement care pathway for quality palliative care.
ABSTRACTObjective:Provision of bereavement support is an essential component of palliative care service delivery. While bereavement support is integral to palliative care, it is typically insufficiently resourced, under-researched, and not systematically applied. Our aim was to develop bereavement standards to assist palliative care services to provide targeted support to family caregivers. We employed a multiple-methods design for our study, which included: (1) a literature review, (2) a survey of palliative care service providers in Australia, (3) interviews with national (Australian) and international experts, (4) key stakeholder workshops, and (5) a modified Delphi-type survey. A total of 10 standards were developed along with a pragmatic care pathway to assist palliative care services with implementation of the standards. The bereavement standards and care pathway constitute a key initiative in the evolution of bereavement support provided by palliative care services. Future endeavors should refine and examine the impact of these standards. Additional research is required to enhance systematic approaches to quality bereavement care.


Toward a multidimensional model for psychotherapy supervision based on developmental stages.
The need for a systematic approach to psychotherapy training in psychiatric education is greater than ever. Most models of psychotherapy supervision focus on supervisee development and ignore developmental issues concerning the other participants in the supervisory triangle. The author reviews theories of psychotherapy supervision and identifies factors that contribute to the dynamic interrelationships among psychiatrists who are supervising psychotherapy, the psychiatry resident supervisees, and their patients as each party progresses through stages of development. A dynamic formulation of psychotherapy supervision based on developmental stages can result in a better understanding of the instructional process, its greater effectiveness, and associated changes in program structure. Further development of a comprehensive dynamic model depends on additional research.


Myosin IIA-related Actomyosin Contractility Mediates Oxidative Stress-induced Neuronal Apoptosis.
Oxidative stress-induced neuronal apoptosis plays an important role in the progression of central nervous system (CNS) diseases. In our study, when neuronal cells were exposed to hydrogen peroxide (H2O2), an exogenous oxidant, cell apoptosis was observed with typical morphological changes including membrane blebbing, neurite retraction and cell contraction. The actomyosin system is considered to be responsible for the morphological changes, but how exactly it regulates oxidative stress-induced neuronal apoptosis and the distinctive functions of different myosin II isoforms remain unclear. We demonstrate that myosin IIA was required for neuronal contraction, while myosin IIB was required for neuronal outgrowth in normal conditions. During H2O2-induced neuronal apoptosis, myosin IIA, rather than IIB, interacted with actin filaments to generate contractile forces that lead to morphological changes. Moreover, myosin IIA knockout using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein-9 nuclease (CRISPR/Cas9) reduced H2O2-induced neuronal apoptosis and the associated morphological changes. We further demonstrate that caspase-3/Rho-associated kinase 1 (ROCK1) dependent phosphorylation of myosin light chain (MLC) was required for the formation of the myosin IIA-actin complex. Meanwhile, either inhibition of myosin II ATPase with blebbistatin or knockdown of myosin IIA with siRNA reversely attenuated caspase-3 activation, suggesting a positive feedback loop during oxidative stress-induced apoptosis. Based on our observation, myosin IIA-actin complex contributes to actomyosin contractility and is associated with the positive feedback loop of caspase-3/ROCK1/MLC pathway. This study unravels the biochemical and mechanistic mechanisms during oxidative stress-induced neuronal apoptosis and may be applicable for the development of therapies for CNS diseases.


In vitro osteogenic differentiation of rat bone marrow cells subcultured with and without dexamethasone.
The aim of our study was to investigate the osteogenic potential of subcultured rat bone marrow cells. Rat bone marrow (RBM) cells were cultured with or without dexamethasone. Subsequently, osteogenic differentiation and expression was studied. When cells were cultured continuously in the presence of dexamethasone, cultures initially showed high alkaline phosphatase expression and abundant mineralization. Expression of differentiation markers decreased with passaging. After cells were passaged three times, no alkaline phosphatase activity and calcification were found. Primary cells cultured without dexamethasone showed low alkaline phosphatase and no calcification, and remained fibroblast-like. When these cells were subcultured in the presence of dexamethasone, the cells did show osteogenic differentiation. Nevertheless, this occurred at a significant lower level than with cells continuously cultured with dexamethasone. In addition, no differentiation was found after second passage. Our results indicate that subcultured undifferentiated RBM cells show osteogenic differentiation after addition of dexamethasone. Expression of alkaline phosphatase and mineralization is higher in cells continuously supplemented with dexamethasone. Still, even when dexamethasone is added continuously, RBM cells loose their osteogenic potential after several passages. Therefore, we conclude that subculture of undifferentiated rat bone marrow cells results in the loss of osteogenic potential of these cells.


Chromic phosphate therapy in carcinoma of the ovary.
Adjuvant therapy with intraperitoneal (IP) chromic phosphate (P-32) instillation in both early and advanced carcinoma of the ovary requires adequate intra-abdominal distribution for maximum therapeutic benefit. Abdominal-pelvic computed tomographic (CT) scanning with water-soluble IP contrast to establish the absence of intra-abdominal adhesions prior to P-32 instillation has not been previously reported. Sixteen patients with carcinoma of the ovary who were offered IP P-32 therapy between January 1988 and December 1992 were retrospectively reviewed. Computed tomographic scans on 13 patients were reevaluated using a modification of Muggia and associates' distribution scoring system. Of the 16 patients, one had a negative second look operation for stage IIIc carcinoma of the ovary. The remaining 15 with stage I and II disease underwent at least a staging laparotomy, bilateral salpingo-oophorectomy, and omentectomy. The mean time from operation to IP P-32 attempt was 28 days with a range of 17 to 90 days. Adequate distribution was considered present in two of the three cases evaluated by technetium scanning and in four of the 13 cases studied by CT scanning. On review, 12 of 13 cases studied by CT scans revealed some areas of IP contrast exclusion and loculation. We suspect that uneven postoperative IP P-32 distribution is common and may be missed by two-dimensional imaging techniques such as technetium scanning or fluoroscopy.


Temperature and pressure effects on C-H abstraction reactions involving compound I and II mimics in aqueous solution.
The presented results cover a comparative mechanistic study on the reactivity of compound (Cpd) I and II mimics of a water-soluble iron(III) porphyrin, [meso-tetrakis(2,4,6-trimethyl-3-sulfonatophenyl)porphinato]iron(III), Fe(III)(TMPS). The acidity of the aqueous medium strongly controls the chemical nature and stability of the high-valent iron(IV) oxo species. Reactivity studies were performed at pH 5 and 10, where the Cpd I and II mimics are stabilized as the sole oxidizing species, respectively. The contributions of ΔH(‡) and ΔS(‡) to the free energy of activation (ΔG(‡)) for the oxidation of 4-methoxybenzaldehyde (4-MB-ald), 4-methoxybenzyl alcohol (4-MB-alc), and 1-phenylethanol (1-PhEtOH) by the Cpd I and II mimics were determined. The relatively large contribution of the ΔH(‡) term in comparison to the -TΔS(‡) term to ΔG(‡) for reactions involving the Cpd II mimic indicates that the oxidation of selected substrates by this oxidizing species is clearly an enthalpy-controlled process. In contrast, different results were found for reactions with application of the Cpd I mimic. Depending on the nature of the substrate, the reaction at room temperature can be entropy-controlled, as found for the oxidation of 4-MB-alc, or enthalpy-controlled, as found for 1-PhEtOH. Importantly, for the first time, activation volumes (ΔV(‡)) for the oxidation of selected substrates by both reactive intermediates could be determined. Positive values of ΔV(‡) were found for reactions with the Cpd II mimic and slightly negative ones for reactions with the Cpd II mimic. The results are discussed in the context of the oxidation mechanism conducted by the Cpd I and II mimics.


Molecular basis of the glycosomal targeting of PEX11 and its mislocalization to mitochondrion in trypanosomes.
PEX19 binding sites are essential parts of the targeting signals of peroxisomal membrane proteins (mPTS). In this study, we characterized PEX19 binding sites of PEX11, the most abundant peroxisomal and glycosomal membrane protein from Trypanosoma brucei and Saccharomyces cerevisiae. TbPEX11 contains two PEX19 binding sites, one close to the N-terminus (BS1) and a second in proximity to the first transmembrane domain (BS2). The N-terminal BS1 is highly conserved across different organisms and is required for maintenance of the steady-state concentration and efficient targeting to peroxisomes and glycosomes in both baker's yeast and Trypanosoma brucei. The second PEX19 binding site in TbPEX11 is essential for its glycosomal localization. Deletion or mutations of the PEX19 binding sites in TbPEX11 or ScPEX11 results in mislocalization of the proteins to mitochondria. Bioinformatic analysis indicates that the N-terminal region of TbPEX11 contains an amphiphilic helix and several putative TOM20 recognition motifs. We show that the extreme N-terminal region of TbPEX11 contains a cryptic N-terminal signal that directs PEX11 to the mitochondrion if its glycosomal transport is blocked.


A Novel Bromophenol Derivative BOS-102 Induces Cell Cycle Arrest and Apoptosis in Human A549 Lung Cancer Cells via ROS-Mediated PI3K/Akt and the MAPK Signaling Pathway.
Bromophenol is a type of natural marine product. It has excellent biological activities, especially anticancer activities. In our study of searching for potent anticancer drugs, a novel bromophenol derivative containing indolin-2-one moiety, 3-(4-(3-([1,4'-bipiperidin]-1'-yl)propoxy)-3-bromo-5-methoxybenzylidene)-N-(4-bromophenyl)-2-oxoindoline-5-sulfonamide (BOS-102) was synthesized, which showed excellent anticancer activities on human lung cancer cell lines. A study of the mechanisms indicated that BOS-102 could significantly block cell proliferation in human A549 lung cancer cells and effectively induce G0/G1 cell cycle arrest via targeting cyclin D1 and cyclin-dependent kinase 4 (CDK4). BOS-102 could also induce apoptosis, including activating caspase-3 and poly (ADP-ribose) polymerase (PARP), increasing the Bax/Bcl-2 ratio, enhancing reactive oxygen species (ROS) generation, decreasing mitochondrial membrane potential (MMP, ΔΨm), and leading cytochrome c release from mitochondria. Further research revealed that BOS-102 deactivated the PI3K/Akt pathway and activated the mitogen-activated protein kinase (MAPK) signaling pathway resulting in apoptosis and cell cycle arrest, which indicated that BOS-102 has the potential to develop into an anticancer drug.


Immediate life support training in an operational environment.
The Immediate Life Support (ILS) course is a 1 day programme designed by the Resuscitation Council (UK) to bridge the gap between Basic Life Support (BLS) and Advanced Life Support (ALS). During the build up to Operation TELIC, the aim was to provide ILS training to all staff who may be involved in the acute care of patients within PCRF ARGUS, and also to members of staff from other units who may be involved in similar situations. In total, 101 staff received ILS training during Op TELIC, all of whom were successful in completing the course.


Dose-Response Models for Eastern US, Western US and Venezuelan Equine Encephalitis Viruses in Mice - Part II: Quantification of the Effects of Host Age on the Dose Response.
Many infectious disease hazards demonstrate higher susceptibility with regards to younger host ages. This trend of increased susceptibility with decreasing host age can also lead to an increased likelihood of mortality, and prolonged/chronic health effects. For quantitative microbial risk assessment (QMRA) modeling, the ability to quantify the effect of host age in the dose response model can allow modelers to account for these effects mechanistically. Additionally, QMRA modelers using age-dependent dose response models can model entire populations within the dose-response itself rather than modeling age ranges using susceptibility factors. This research developed host-age dependent exponential and beta Poisson dose response models for Eastern, Western and Venezuelan encephalitis viruses (EEV, WEV and VEV respectively) for two routes - intracranial and intraperitoneal. Improvement in fit was statistically tested as a means of assessing the benefit of including age dependency into the dose response models. EEV demonstrated improvement in fit using host-age dependency only for the exponential model except for intracranial exposure. EEV demonstrated an improvement in fit when using age dependency in the beta Poisson dose response model for both exposure routes. VEV demonstrated an improvement in fit using age dependency for both exposure routes. WEV demonstrated an improvement in fit for intracranial exposure, but neither of the age dependent dose-response models provided a good fit for WEV intraperitoneal exposure.


Digital radiography for target point evaluation in stereotactic neurosurgery.
Today, the usual target point evaluation during stereotactic operations using planar X-ray films can be supplemented by digital video storage. The picture-delivering system is a digital X-ray video chain which replaces the X-ray film. Evaluation of target point coordinates as well as parameters of electrode approach and positioning are determined by a microcomputer. Such an inexpensive system is described.


Subunit contributions to phosphorylation-dependent modulation of bovine rod cyclic nucleotide-gated channels.
Cyclic nucleotide-gated (CNG) channels in rod photoreceptors transduce a decrease in cGMP into hyperpolarization during the light response. Insulin-like growth factor-1 (IGF-1) increases light responses by increasing the cGMP sensitivity of CNG channels, an event mediated by a protein tyrosine phosphatase. Native rod CNG channels are heteromultimers, composed of three CNGA1 subunits and one CNGB1 subunit. Previous studies on heterologously expressed rod CNG channels show that a specific tyrosine in the CNGA1 subunit (Y498) is required for modulation by protein tyrosine phosphatases, protein tyrosine kinases and IGF-1. Here we show that the CNGB1 subunit contains a specific tyrosine (Y1097) that is important for modulation of heteromeric channels by tyrosine phosphorylation. Direct biochemical measurements demonstrate 32P-labelling of CNGA1Y498 and CNGB1Y1097. Replacement of either Y498 of CNGA1 or Y1097 of CNGB1 with phenylalanine reduces modulation, and removal of both tyrosines eliminates modulation. Unlike CNGA1, CNGB1 does not exhibit activity dependence of modulation by tyrosine phosphorylation. Hence both CNGA1 and CNGB1 subunits contribute to phosphorylation-dependent modulation of rod CNG channels, but the phosphorylation states of the two subunits are regulated in different ways.


Systemic fungal infections: diagnosis and treatment. I. Paracoccidioidomycosis.
Paracoccidioidomycosis is the designation for the disease caused by the dimorphic fungus Paracoccidioides brasiliensis. It is a disease of great importance in Latin America and constitutes an important diagnosis for consideration in patients from endemic areas who have pulmonary or systemic disease compatible with fungal infection. In the laboratory, basic work has uncovered important interactions between the fungus and the human sex steroid system. These studies may have important implications in our approach to this and other infectious diseases.


SR proteins control a complex network of RNA-processing events.
SR proteins are a well-conserved class of RNA-binding proteins that are essential for regulation of splice-site selection, and have also been implicated as key regulators during other stages of RNA metabolism. For many SR proteins, the complexity of the RNA targets and specificity of RNA-binding location are poorly understood. It is also unclear if general rules governing SR protein alternative pre-mRNA splicing (AS) regulation uncovered for individual SR proteins on few model genes, apply to the activity of all SR proteins on endogenous targets. Using RNA-seq, we characterize the global AS regulation of the eight Drosophila SR protein family members. We find that a majority of AS events are regulated by multiple SR proteins, and that all SR proteins can promote exon inclusion, but also exon skipping. Most coregulated targets exhibit cooperative regulation, but some AS events are antagonistically regulated. Additionally, we found that SR protein levels can affect alternative promoter choices and polyadenylation site selection, as well as overall transcript levels. Cross-linking and immunoprecipitation coupled with high-throughput sequencing (iCLIP-seq), reveals that SR proteins bind a distinct and functionally diverse class of RNAs, which includes several classes of noncoding RNAs, uncovering possible novel functions of the SR protein family. Finally, we find that SR proteins exhibit positional RNA binding around regulated AS events. Therefore, regulation of AS by the SR proteins is the result of combinatorial regulation by multiple SR protein family members on most endogenous targets, and SR proteins have a broader role in integrating multiple layers of gene expression regulation.


Visual perception during mirror-gazing at one's own face in patients with depression.
In normal observers, gazing at one's own face in the mirror for a few minutes, at a low illumination level, produces the apparition of strange faces. Observers see distortions of their own faces, but they often see hallucinations like monsters, archetypical faces, faces of relatives and deceased, and animals. In this research, patients with depression were compared to healthy controls with respect to strange-face apparitions. The experiment was a 7-minute mirror-gazing test (MGT) under low illumination. When the MGT ended, the experimenter assessed patients and controls with a specifically designed questionnaire and interviewed them, asking them to describe strange-face apparitions. Apparitions of strange faces in the mirror were very reduced in depression patients compared to healthy controls. Depression patients compared to healthy controls showed shorter duration of apparitions; minor number of strange faces; lower self-evaluation rating of apparition strength; lower self-evaluation rating of provoked emotion. These decreases in depression may be produced by deficits of facial expression and facial recognition of emotions, which are involved in the relationship between the patient (or the patient's ego) and his face image (or the patient's bodily self) that is reflected in the mirror.


BIOFACE: A Prospective Study of Risk Factors, Cognition, and Biomarkers in a Cohort of Individuals with Early-Onset Mild Cognitive Impairment. Study Rationale and Research Protocols.
Mild cognitive impairment (MCI) due to Alzheimer's disease (AD) diagnosis is based on cerebrospinal fluid (CSF) or neuroimaging biomarkers. Currently, non-invasive and inexpensive blood-based biomarkers are being investigated, such as neuronal-derived plasma exosomes (NPEs). Neuroinflammation and early vascular changes have been described in AD pathogenesis and can be traced in plasma and NPEs. However, they have not been studied in early onset MCI (EOMCI). To describe the rationale, design, and baseline characteristics of the participants from the BIOFACE cohort, a two-year observational study on EOMCI conducted at Fundació ACE. The study goal is to characterize the different phenotypes from a clinical, neuropsychological, and biomarker point of view and to investigate the CSF and plasma proteomics as well as the role of NPEs as early biomarkers of AD. Participants underwent extended neurological and neuropsychological batteries, multimodal biomarkers including brain MRI, blood, saliva, CSF, anthropometric, and neuro-ophthalmological examinations. Ninety-seven patients with EOMCI were recruited. 59.8%were women. Mean age at symptom onset was 57 years; mean MMSE was 28. First degree and presenile family history of dementia was present in 60.8%and 15.5%, respectively. Depressive and anxiety disorders along with vascular risk factors were the most frequent comorbidities. 29%of participants were APOE ɛ4 carriers, and 67%showed a CSF normal ATN profile. BIOFACE is a two-year study of clinical, cognition, and biomarkers that will shed light on the physiopathology and the potential utility of plasma and NPEs as non-invasive early diagnostic and prognostic biomarkers in people younger than 65 years.


Clock proteins and training modify exercise capacity in a daytime-dependent manner.
Exercise and circadian biology are closely intertwined with physiology and metabolism, yet the functional interaction between circadian clocks and exercise capacity is only partially characterized. Here, we tested different clock mutant mouse models to examine the effect of the circadian clock and clock proteins, namely PERIODs and BMAL1, on exercise capacity. We found that daytime variance in endurance exercise capacity is circadian clock controlled. Unlike wild-type mice, which outperform in the late compared with the early part of their active phase, PERIODs- and BMAL1-null mice do not show daytime variance in exercise capacity. It appears that BMAL1 impairs and PERIODs enhance exercise capacity in a daytime-dependent manner. An analysis of liver and muscle glycogen stores as well as muscle lipid utilization suggested that these daytime effects mostly relate to liver glycogen levels and correspond to the animals' feeding behavior. Furthermore, given that exercise capacity responds to training, we tested the effect of training at different times of the day and found that training in the late compared with the early part of the active phase improves exercise performance. Overall, our findings suggest that clock proteins shape exercise capacity in a daytime-dependent manner through changes in liver glycogen levels, likely due to their effect on animals' feeding behavior.


Elucidating the mechanism behind the infrared spectral features and dynamics observed in the carbonyl stretch region of organic carbonates interacting with lithium ions.
Ultrafast infrared spectroscopy has become a very important tool for studying the structure and ultrafast dynamics in solution. In particular, it has been recently applied to investigate the molecular interactions and motions of lithium salts in organic carbonates. However, there has been a discrepancy in the molecular interpretation of the spectral features and dynamics derived from these spectroscopies. Hence, the mechanism behind spectral features appearing in the carbonyl stretching region was further investigated using linear and nonlinear spectroscopic tools and the co-solvent dilution strategy. Lithium perchlorate in a binary mixture of dimethyl carbonate (DMC) and tetrahydrofuran was used as part of the dilution strategy to identify the changes of the spectral features with the number of carbonates in the first solvation shell since both solvents have similar interaction energetics with the lithium ion. Experiments showed that more than one carbonate is always participating in the lithium ion solvation structures, even at the low concentration of DMC. Moreover, temperature-dependent study revealed that the exchange of the solvent molecules coordinating the lithium ion is not thermally accessible at room temperature. Furthermore, time-resolved IR experiments confirmed the presence of vibrationally coupled carbonyl stretches among coordinated DMC molecules and demonstrated that this process is significantly altered by limiting the number of carbonate molecules in the lithium ion solvation shell. Overall, the presented experimental findings strongly support the vibrational energy transfer as the mechanism behind the off-diagonal features appearing on the 2DIR spectra of solutions of lithium salt in organic carbonates.


Autogenous bone grafts and bone substitutes--tools and techniques: I. A 20,000-case experience in maxillofacial and craniofacial surgery.
Alloplastic bone substitutes can be used to alter facial contour. In contrast, autogenous bone grafts have a successful 80-year history of restoring facial contour as well as the basic functional support of the craniofacial skeleton. The traditional procedures for harvesting and using autogenous bone grafts are not obsolete. During the past 30 years, the techniques have been refined and new sources have been found, such as calvarial grafts. New tools were required and have been designed to make harvesting of grafts easier and faster for the surgeon and safer and less expensive for the patient. Four short articles under the heading of "Techniques and Tools" are presented addressing the harvesting of (1) iliac, (2) costal, (3) tibial, and (4) calvarial grafts. These articles are based on the experience of six surgeons using the same technique and instruments in more than 20,000 autogenous bone grafting procedures. (These figures represent the group experience as of 2001. Since then, one of the junior coauthors has retired, but the remaining five continue to harvest autogenous bone grafts on a regular basis. So, the group experience as of 2004 is in the range of 23,000 procedures). The-senior surgeon's experience of 9500 procedures spans a period of 50 years (from 1946 to 1996). For the other surgeons (10,500 procedures combined), the collection period was 25 years (from 1975 to 2000).


Macrophages and Urokinase Plasminogen Activator Receptor System in Multiple Myeloma: Case Series and Literature Review.
The microenvironment plays an essential role in multiple myeloma (MM) development, progression, cell proliferation, survival, immunological escape, and drug resistance. Mesenchymal stromal cells and macrophages release tolerogenic cytokines and favor anti-apoptotic signaling pathway activation, while the urokinase plasminogen activator receptor (uPAR) system contributes to migration through an extracellular matrix. Here, we first summarized the role of macrophages and the uPAR system in MM pathogenesis, and then we reported the potential therapeutic effects of uPAR inhibitors in a case series of primary MM-derived adherent cells. Our preliminary results showed that after uPAR inhibitor treatments, interleukein-6 (mean ± SD, 8734.95 ± 4169.2 pg/mL vs. 359.26 ± 393.8 pg/mL, pre- vs. post-treatment; p = 0.0012) and DKK-1 levels (mean ± SD, 7005.41 ± 6393.4 pg/mL vs. 61.74 ± 55.2 pg/mL, pre- vs. post-treatment; p = 0.0043) in culture medium were almost completely abolished, supporting further investigation of uPAR blockade as a therapeutic strategy for MM treatment. Therefore, uPAR inhibitors could exert both anti-inflammatory and pro-immunosurveillance activity. However, our preliminary results need further validation in additional in vitro and in vivo studies.


A giant congenital orbital tumor: an unusual presentation of retinoblastoma.
We report a case of an unusual giant congenital tumor presenting in a newborn infant as a large exophytic mass emerging from the left orbit. After enucleation orbital recurrence developed within 14 days. No anti-tumor treatment was given and the child died at the age of 4 weeks. The histopathological and cytogenetical analysis of the tumor is presented. The tumor was diagnosed as a retinoblastoma but we could not exclude the possibility of a neuroblastoma.


The UK Out of Hospital Cardiac Arrest Outcome (OHCAO) project.
Reducing premature death is a key priority for the UK National Health Service (NHS). NHS Ambulance services treat approximately 30 000 cases of suspected cardiac arrest each year but survival rates vary. The British Heart Foundation and Resuscitation Council (UK) have funded a structured research programme--the Out of Hospital Cardiac Arrest Outcomes (OHCAO) programme. The aim of the project is to establish the epidemiology and outcome of OHCA, explore sources of variation in outcome and establish the feasibility of setting up a national OHCA registry. This is a prospective observational study set in UK NHS Ambulance Services. The target population will be adults and children sustaining an OHCA who are attended by an NHS ambulance emergency response and where resuscitation is attempted. The data collected will be characterised broadly as system characteristics, emergency medical services (EMS) dispatch characteristics, patient characteristics and EMS process variables. The main outcome variables of interest will be return of spontaneous circulation and medium-long-term survival (30 days to 10-year survival). Ethics committee permissions were gained and the study also has received approval from the Confidentiality Advisory Group Ethics and Confidentiality committee which provides authorisation to lawfully hold identifiable data on patients without their consent. To identify the key characteristics contributing to better outcomes in some ambulance services, reliable and reproducible systems need to be established for collecting data on OHCA in the UK. Reports generated from the registry will focus on data completeness, timeliness and quality. Subsequent reports will summarise demographic, patient, process and outcome variables with aim of improving patient care through focus quality improvement initiatives.


ExSTraCS 2.0: Description and Evaluation of a Scalable Learning Classifier System.
Algorithmic scalability is a major concern for any machine learning strategy in this age of 'big data'. A large number of potentially predictive attributes is emblematic of problems in bioinformatics, genetic epidemiology, and many other fields. Previously, ExS-TraCS was introduced as an extended Michigan-style supervised learning classifier system that combined a set of powerful heuristics to successfully tackle the challenges of classification, prediction, and knowledge discovery in complex, noisy, and heterogeneous problem domains. While Michigan-style learning classifier systems are powerful and flexible learners, they are not considered to be particularly scalable. For the first time, this paper presents a complete description of the ExS-TraCS algorithm and introduces an effective strategy to dramatically improve learning classifier system scalability. ExSTraCS 2.0 addresses scalability with (1) a rule specificity limit, (2) new approaches to expert knowledge guided covering and mutation mechanisms, and (3) the implementation and utilization of the TuRF algorithm for improving the quality of expert knowledge discovery in larger datasets. Performance over a complex spectrum of simulated genetic datasets demonstrated that these new mechanisms dramatically improve nearly every performance metric on datasets with 20 attributes and made it possible for ExSTraCS to reliably scale up to perform on related 200 and 2000-attribute datasets. ExSTraCS 2.0 was also able to reliably solve the 6, 11, 20, 37, 70, and 135 multiplexer problems, and did so in similar or fewer learning iterations than previously reported, with smaller finite training sets, and without using building blocks discovered from simpler multiplexer problems. Furthermore, ExS-TraCS usability was made simpler through the elimination of previously critical run parameters.


A chemical method to enrich RNA by molecules having 5'-terminal triphosphate groups.
A method is proposed to enrich RNA molecules having a 5'-terminal triphosphate group. The method is based upon selective chemical modification of 5'-triphosphate groups by an antigen-containing amine followed by affinity chromatography on an absorbent loaded with antibodies specific to this antigen. A purification factor up to 37 may be achieved.


Intensive Care Unit Admission and Death Rates of Infants Admitted With Respiratory Syncytial Virus Lower Respiratory Tract Infection in Mexico.
Respiratory syncytial virus (RSV) is the most common etiology for acute respiratory infection hospital admissions in young children. Case fatality rates for hospitalized patients range between 0% and 3.4%. Recent reports indicate that deaths associated with RSV are uncommon in developed countries. However, the role of this virus as a current cause of mortality in other countries requires further examination. Children with RSV infection admitted between May 2003 and December 2014 to a level 2 specialty hospital in Mexico were included in this analysis. Underlying risk factors, admission to the intensive care unit (ICU) and condition on discharge were assessed to determine the ICU admission and death rates associated to RSV infection. We analyzed data of 1153 patients with RSV infection in whom information regarding underlying illnesses and discharge status was available. Sixty patients (5.2 %) were admitted to the ICU and 12 (1.04 %) died. Relevant underlying conditions were present in 320 (27.7%) patients. Infants with underlying respiratory disorders (excluding asthma) and a history of prematurity had high ICU admission rates (17.1% and 13.8%, respectively). Mortality rates were highest for infants with respiratory disease (excluding asthma) (7.3%), cardiovascular diseases (5.9%) and neurologic disorders (5.3%). The ICU admission and death rates were higher in infants <6 months of age than in other age groups. The ICU admission rate and mortality rate in Mexican infants hospitalized with RSV infection were 5.2% and 1%, respectively. Mortality rates were high in infants with respiratory, cardiovascular and neurologic disorders.


A glucuronoxylan-specific xylanase from a new Paenibacillus favisporus strain isolated from tropical soil of Brazil.
A new xylanolytic strain, Paenibacillus favisporus CC02-N2, was isolated from sugarcane plantation fields in Brazil. The strain had a xylan-degrading system with multiple enzymes, one of which, xylanase Xyn30A, was identified and characterized. The enzyme is a single-domain xylanase belonging to family 30 of the glycosyl hydrolases (GH30). Xyn30A shows high activity on glucuronoxylans, with a Vmax of 267.2 U mg⁻¹, a Km of 4.0 mg/ml, and a kcat of 13,333 min⁻¹ on beechwood xylan, but it does not hydrolyze arabinoxylans. The three-dimensional structure of Xyn30A consists of a common (β/α)8 barrel linked to a side-chain-associated β-structure, similar to previously characterized GH30 xylanases. The hydrolysis products from glucuronoxylan were methylglucuronic-acid-substituted xylooligomers (acidic xylooligosaccharides). The enzyme bound to insoluble xylan but not to crystalline cellulose. Our results suggest a specific role for Xyn30A in xylan biodegradation in natural habitats. The enzyme is a good candidate for the production of tailored xylooligosaccharides for use in the food industry and in the biotechnological transformation of biomass.


Effect of shear stress on structure and function of polyplex micelles from poly(ethylene glycol)-poly(l-lysine) block copolymers as systemic gene delivery carrier.
Structural stability of polyplex micelles (PMs), prepared from plasmid DNA (pDNA) and poly(ethylene glycol)-b-poly(l-lysine) block catiomer (PEG-PLys), was evaluated in terms of their resistance against shear stress. When exposed to shear stress at magnitudes typically present in the blood stream, structural deterioration was observed in PMs owing to the partial removal of PEG-PLys strands. Eventually, impaired PEG coverage of the polyplex core led to accelerated degradation by nucleases, implying that structural deterioration by shear stress in blood stream may be a major cause of rapid clearance of PMs from blood circulation. To address this issue, introduction of disulfide crosslinking into the PM core was shown to be an efficient strategy, which successfully mitigated unfavorable effects of shear stress. Furthermore, improved in vivo blood retention profile and subsequently enhanced antitumor efficacy in systemic treatment of pancreatic adenocarcinoma were confirmed for the crosslinked PMs loaded with pDNA encoding an anti-angiogenic protein, suggesting that high stability under the shear stress during blood circulation may be a critical factor in systemically applicable gene delivery systems.


Age of Onset of Functional Constipation.
In a review of 538 children with functional constipation, we analyzed ages of presentation and onset, symptom duration, and behavioral/developmental problems. We divided the subjects into quartiles (Q1-Q4) based on age of onset. Median onset age was 2.3 years. The oldest group had the shortest symptom duration before referral at 1.8 ± 1.8 years (compared with Q3 to Q1, P = 0.039, P = 0.001, P < 0.001, respectively). Of the Q4 subjects, 22% had a behavioral/developmental problem (P < 0.001 compared with Q1-Q3). We conclude that most children develop functional constipation as infants and toddlers, but those with later onset are more likely to have behavioral/developmental issues and see a specialist sooner.


The effect of chitosan coating combined with cold plasma on the quality and safety of pistachio during storage.
Pistachios are one of the most important agricultural and export products of Iran. Fresh pistachio fruit has soft skin, is highly perishable, and therefore has a short life after harvesting, which has made traders and consumers have a great desire to increase the shelf life of this product. For this purpose, in this study, the effect of different concentrations of chitosan as an edible coating (0.5 and 1.5% w/v) and the duration of cold plasma treatment (60 and 120 s) were investigated during 180 days of pistachio storage. The effect of treatments on the shelf life of pistachio fruit was evaluated by determining moisture content, color components, peroxide value, total mold and yeast, hardness, aflatoxin content, and sensory evaluations. The results showed that the treatment with 1.5% chitosan coating and 120 s of cold plasma treatment preserved the hardness of the pistachio and the color indices in the best way (p < .05). Also, this treatment had the minimum number of peroxide, aflatoxin, and mold and yeast counts during the storage time. The treatments with chitosan coating and under plasma application did not cause any unpleasant odor or taste during the storage time. In conclusion, according to the results of this research, it was determined that the simultaneous use of chitosan coating and cold plasma treatment can potentially be used as a new approach for commercial applications and the export of fresh pistachios.


Research hotspots and trends in retinopathy of prematurity from 2003 to 2022: a bibliometric analysis.
In order to understand the research hotspots and trends in the field of retinopathy of prematurity (ROP), our study analyzed the relevant publications from 2003 to 2022 by using bibliometric analysis. The Citespace 6.2.R3 system was used to analyze the publications collected from the Web of Science Core Collection (WoSCC) database. In total, 4,957 publications were included in this study. From 2003 to 2022, the number of publications gradually increased and peaked in 2022. The United States was the country with the most publications, while Harvard University was the most productive institution. The top co-cited journal PEDIATRICS is published by the United States. Author analysis showed that Hellström A was the author with the most publications, while Good WV was the top co-cited author. The co-citation analysis of references showed seven major clusters: genetic polymorphism, neurodevelopmental outcome, threshold retinopathy, oxygen-induced retinopathy, low birth weight infant, prematurity diagnosis cluster and artificial intelligence (AI). For the citation burst analysis, there remained seven keywords in their burst phases until 2022, including ranibizumab, validation, trends, type 1 retinopathy, preterm, deep learning and artificial intelligence. Intravitreal anti-vascular endothelial growth factor therapy and AI-assisted clinical decision-making were two major topics of ROP research, which may still be the research trends in the coming years.