Live Imaging of Shoot Meristems on an Inverted Confocal Microscope Using an Objective Lens Inverter Attachment.
Live imaging of above ground meristems can lead to new insights in plant development not possible from static imaging of fixed tissue. The use of an upright confocal microscope offers several technical and biological advantages for live imaging floral or shoot meristems. However, many departments and core facilities possess only inverted confocal microscopes and lack the funding for an additional upright confocal microscope. Here we show that imaging of living apical meristems can be performed on existing inverted confocal microscopes with the use of an affordable and detachable InverterScope accessory.


Multidimensionality and bipolarity of a Spanish version of Warr's (1990) well-being measures.
The validity and reliability of the Spanish version of the 2 scales measuring work-related well-being proposed by P. Warr (1990; Anxiety-Contentment and Depression-Enthusiasm) were examined in a cross-sectional questionnaire study among 299 Spanish factory employees. Two methods suggested by Warr (1990) to control the response bias (acquiescent response set) were used. The 1st method involved separate analyses for the 6 positive and 6 negative items that make up the 2 scales. The 2nd was a series of 4 principal component analyses with oblique rotation on the estimated response set controlled in partial correlation between each item. The results support the existence of these 2 independent scales for measuring work-related psychological well-being as hypothesised by Warr (1990). However, 2 main problems appear: the cross-loading of the items contented and cheerful and the fact that only I factor appears in the pre-rotated solution. Possible explanations for these results are discussed.


Chronic urticaria following penicillin therapy.
Urticaria developed in six patients three to ten days after the intramuscular injection of penicillin, and the condition persisted for from three to fifteen months. The suffering, disability and expense that resulted from the injection indicates that physicians should make certain that the patient who receives penicillin has a true indication for its use.


GEOGRAPHIC VARIATION IN FEMALE PREFERENCES FOR MALE TRAITS IN POECILIA RETICULATA.
We examined the preferences of female guppies (Poecilia reticulata) from 11 localities in Trinidad with respect to male color-pattern elements, body shape and size, and overall color and brightness contrast. Females are on average more attracted to males from their own population than from alien populations, and populations appear to vary in the criteria used in female choice. Multiple-regression analysis suggests that mate-preference criteria vary among localities in intensity, sign, and the number of traits used. Although preference estimators and color-pattern parameters are unique to each population, only orange, black, and color contrast showed a correlation between degree of male trait and degree of preference for that trait. There is a clear effect of water color and a possible effect of predation intensity. The results are discussed in light of various models of sexual selection and the early stages of speciation.


In search of acceptable alternatives to the murine histamine sensitisation test (HIST): what is possible and practical?
The 'International Workshop on Alternatives to the Murine Histamine Sensitization Test for Acellular Pertussis Vaccines: In Search of Acceptable Alternatives to the Murine Histamine Sensitization Test (HIST): What is Possible and Practical?' was held on 4 and 5 March 2015 in London, United Kingdom. Participants discussed the results of the data generated from an international collaborative study (BSP114 Phase 2) sponsored by the European Directorate for the Quality of Medicines & Health Care (EDQM) to determine if a modified Chinese hamster ovary (CHO) cell-based clustering assay is a suitable alternative to replace HIST. Workshop participants agreed that protocol transferability demonstrated in the collaborative study indicates that a standardised CHO cell assay is adequate for measuring pure PTx in reference preparations. However, vaccine manufacturers would still need to demonstrate that the method is valid to detect or measure residual PTx in their specific adjuvanted products. The 2 modified CHO cell protocols included in the study (the Direct and the Indirect Methods) deserve further consideration as alternatives to HIST. Using the CHO cell assay, an in vitro alternative, for acellular pertussis (aP) vaccine batch release testing would reduce the number of animals used for aP vaccine safety testing. A strategic, stepwise adoption plan was proposed, in which the alternative test would be used for release purposes first, and then, once sufficient confidence in its suitable performance has been gained, its use would be extended to stability testing.


Profiling of the perturbed metabolomic state of mouse spleen during acute and chronic toxoplasmosis.
Toxoplasma gondii, a common opportunistic protozoan, is a leading cause of illness and mortality among immunosuppressed individuals and during congenital infections. Current therapeutic strategies for toxoplasmosis are not fully effective at curtailing disease progression in these cases. Given the parasite ability to influence host immunity and metabolism, understanding of the metabolic alterations in the host's immune organs during T. gondii infection may enhance the understanding of the molecular mechanisms that define the pathophysiology of T. gondii infection. We investigated the global metabolic changes in the spleen of BALB/c mice at early and late stage of infection with T. gondii using LC-MS/MS-based metabolomics. Multivariate data analysis methods, principal components analysis (PCA) and partial least squares discriminant analysis (PLS-DA), were used to identify metabolites that are influenced by T. gondii infection. Multivariate analyses clearly separated the metabolites of spleen of infected and control mice. A total of 132 differential metabolites were identified, 23 metabolites from acutely infected versus control mice and 109 metabolites from chronically infected versus control mice. Lipids, hormones, lactones, acids, peptides, antibiotics, alkaloids and natural toxins were the most influenced chemical groups. There were 12 shared differential metabolites between acutely infected versus control mice and chronically infected versus control mice, of which 4,4-Dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol was significantly upregulated and ubiquinone-8 was significantly downregulated. Major perturbed metabolic pathways included primary bile acid biosynthesis, steroid hormone biosynthesis, biotin metabolism, and steroid biosynthesis, with arachidonic acid metabolism being the most significantly impacted pathway. These metabolic changes suggest a multifactorial nature of the immunometabolic responses of mouse spleen to T. gondii infection. This study demonstrated that T. gondii infection can cause significant metabolomic alterations in the spleen of infected mice. These findings provide new insights into the molecular mechanisms that underpin the pathogenesis of T. gondii infection.


Community-consumerism: negotiating risk in online drug communities.
This study explores the social organisation of risk within online drug-related communities. Drawing on in-depth interviews with participants from two Norwegian Internet drug forums, the paper illustrates how participation in such forums influenced notions of risk, and how it supported notions of participants as being informed, responsible and empowered. First, the forums facilitated an easy exchange of user-generated drug information, which helped members present themselves as informed and competent. Second, members used the communal resources on the forums to negotiate their drug-using identities, in which they resisted stigma and argued for a responsible drug-using identity. Third, the social inclusion and sense of community within the forums formed the basis for collective support, which helped empower those involved. Conceptualised as community-consumerism, these findings highlight the social mechanisms involved in the information self-sufficiency and decentralisation of authority on the Internet, in which members created an alternative frame of reference for drug use and associated health. The concept of community-consumerism offers a perspective on the social organisation of risk within online communities and ought to be of relevance for future studies on online health-related discourses, not only those related to drugs.


Immunoreactive Proteins in the Esophageal Gland Cells of Anisakis Simplex Sensu Stricto Detected by MALDI-TOF/TOF Analysis.
In plant and animal nematode parasites, proteins derived from esophageal gland cells have been shown to be important in the host-nematodes relationship but little is known about the allergenic potential of these proteins in the genus Anisakis. Taking into account the increase of anisakiasis and allergies related to these nematodes, immunoreactive properties of gland cell proteins were investigated. Two hundred ventricles were manually dissected from L3 stage larvae of Aniskakis simplex s.s. to allow direct protein analysis. Denaturing gel electrophoresis followed by monochromatic silver staining which revealed the presence of differential (enriched) proteins when compared to total nematode extracts. Such comparison was performed by means of 1D and 2D electrophoresis. Pooled antisera from Anisakis spp.-allergic patients were used in western blots revealing the presence of 13 immunoreactive bands in the ventricular extracts in 1D, with 82 spots revealed in 2D. The corresponding protein bands and spots were excised from the silver-stained gel and protein assignation was made by MALDI-TOF/TOF. A total of 13 (including proteoforms) were unambiguously identified. The majority of these proteins are known to be secreted by nematodes into the external environment, of which three are described as being major allergens in other organisms with different phylogenetic origin and one is an Anisakis simplex allergen.


THERAPY OF ENDOCRINE DISEASE: Amylin and calcitonin - physiology and pharmacology.
Type 2 diabetes is a common manifestation of metabolic dysfunction due to obesity and constitutes a major burden for modern health care systems, in concert with the alarming rise in obesity worldwide. In recent years, several successful pharmacotherapies improving glucose metabolism have emerged and some of these also promote weight loss, thus, ameliorating insulin resistance. However, the progressive nature of type 2 diabetes is not halted by these new anti-diabetic pharmacotherapies. Therefore, novel therapies promoting weight loss further and delaying diabetes progression are needed. Amylin, a beta cell hormone, has satiating properties and also delays gastric emptying and inhibits postprandial glucagon secretion with the net result of reducing postprandial glucose excursions. Amylin acts through the six amylin receptors, which share the core component with the calcitonin receptor. Calcitonin, derived from thyroid C cells, is best known for its role in humane calcium metabolism, where it inhibits osteoclasts and reduces circulating calcium. However, calcitonin, particularly of salmon origin, has also been shown to affect insulin sensitivity, reduce the gastric emptying rate and promote satiation. Preclinical trials with agents targeting the calcitonin receptor and the amylin receptors, show improvements in several parameters of glucose metabolism including insulin sensitivity and some of these agents are currently undergoing clinical trials. Here, we review the physiological and pharmacological effects of amylin and calcitonin and discuss the future potential of amylin and calcitonin-based treatments for patients with type 2 diabetes and obesity.


Adaptation and validation of the Spanish version of the Social Comparison Scale in chronic illness patients.
This study examined the Social Comparison Scale in Spanish chronic illness context and analyzed its psychometric properties. The article presented the results of two studies. The first explored the test's dimensional structure in a sample of 202 patients in a range of several chronic illnesses. The second study examined the instrument's structure in a sample of 186 patients with specific chronic illness. The results replicated the original structure of the scale and proved to be valid for use with optimum reliability.


Cortical atrophy and personality in multiple sclerosis.
Although the cognitive disorder of multiple sclerosis (MS) is well characterized, little is known about personality changes that may occur in this disease. There are reliable personality tests available for research in neurological disease, based on the well-known Five Factor Model. Preliminary research suggests that cognitively impaired MS patients exhibit elevation in Neuroticism, and diminution in Extraversion, Agreeableness, and Conscientiousness, as do patients with Alzheimer's disease. We predicted that these characteristics would be associated with lower neocortical volume. We studied 44 patients using brain MRI and the NEO Five-Factor Inventory. Regression models controlling for T2 lesion volume, depression, and cognitive dysfunction revealed significant correlation between cortical atrophy and reduction in Extraversion and Conscientiousness. Discrepancies between patient- and informant-reports were found, and overreporting of high Openness and Conscientiousness among patients was associated with lower neocortical volume. A final regression model accounting for depression, cognitive function, and personality accounted for 38% of the variance in neocortical volume. These findings suggest that cortical atrophy in MS is associated with adverse impact on personality, although longitudinal research is needed to test this hypothesis.


Regularization and a general linear model for event-related potential estimation.
The usual event-related potential (ERP) estimation is the average across epochs time-locked on stimuli of interest. These stimuli are repeated several times to improve the signal-to-noise ratio (SNR) and only one evoked potential is estimated inside the temporal window of interest. Consequently, the average estimation does not take into account other neural responses within the same epoch that are due to short inter stimuli intervals. These adjacent neural responses may overlap and distort the evoked potential of interest. This overlapping process is a significant issue for the eye fixation-related potential (EFRP) technique in which the epochs are time-locked on the ocular fixations. The inter fixation intervals are not experimentally controlled and can be shorter than the neural response's latency. To begin, the Tikhonov regularization, applied to the classical average estimation, was introduced to improve the SNR for a given number of trials. The generalized cross validation was chosen to obtain the optimal value of the ridge parameter. Then, to deal with the issue of overlapping, the general linear model (GLM), was used to extract all neural responses inside an epoch. Finally, the regularization was also applied to it. The models (the classical average and the GLM with and without regularization) were compared on both simulated data and real datasets from a visual scene exploration in co-registration with an eye-tracker, and from a P300 Speller experiment. The regularization was found to improve the estimation by average for a given number of trials. The GLM was more robust and efficient, its efficiency actually reinforced by the regularization.


Is there a place for reflective practice in the nursing curriculum?
Within this paper a review of the literature on the area of reflection and reflective practice is presented. Lecturers at Trinity College Dublin currently face a dilemma with regard to including these concepts in the nursing curricula. They are faced on the one hand with national recommendations for the use of reflection and reflective practice, and on the other with personal and professional concerns about the practical, legal, ethical and moral implications raised in teaching and assessing this subj ect. The literature reveals that there is little consensus of opinion regarding the exact nature of reflection. There is also little empirical evidence to support the benefits of teaching and assessing of reflection or reflective practice from either an educational or clinical perspective. This, together with the practical difficulties of teaching and assessing reflection, may negate against the incorporation of reflection and reflective practice into the nursing curriculum. It is suggested that, rathe r than isolate reflection as a distinct topic, that the skills of critical thinking, problem solving, self-awareness and analysis be developed within the curriculum.


Quality Control and Basics of Sterile Compounding: Attempting to Avoid Out-of-Specification Results and Patient Harm in Sterile Compounding.
What do you do when something goes wrong in your cleanroom? What do you do when a sterility test fails and what are the implications of such a failure? What should you do when your environmental monitoring or personnel monitoring provides results that exceed your action levels? A root cause analysis is needed in any situation where action levels are exceeded or tests are out of specification. However, should the questions be what can be done to prevent these occurrences, and how can the development be noticed before they occur? In this article, we will discuss going beyond the recommendations of United States Pharmacopeia Chapter <797> to attempt to build a program that limits the possibility of contamination through monitoring of the facility, personnel, and procedures in aseptic processing. We will also discuss other resources published by the U.S. Food and Drug Administration that address monitoring of sterile compounding and aseptic processing.


A review of foot finite element modelling for pressure ulcer prevention in bedrest: Current perspectives and future recommendations.
Pressure ulcers (PUs) are a major public health challenge, having a significant impact on healthcare service and patient quality of life. Computational biomechanical modelling has enhanced PU research by facilitating the investigation of pressure responses in subcutaneous tissue and skeletal muscle. Extensive work has been undertaken on PUs on patients in the seated posture, but research into heel ulcers has been relatively neglected. The aim of this review was to address the key challenges that exist in developing an effective FE foot model for PU prevention and the confusion surrounding the wide range of outputs reported. Nine FE foot studies investigating heel ulcers in bedrest were identified and reviewed. Six studies modelled the posterior part of the heel, two included the calf and foot, and one modelled the whole body. Due to the complexity of the foot anatomy, all studies involved simplification or assumptions regarding parts of the foot structure, boundary conditions and material parameters. Simulations aimed to understand better the stresses and strains exhibited in the heel soft tissues of the healthy foot. The biomechanical properties of soft tissue derived from experimental measurements are critical for developing a realistic model and consequently guiding clinical decisions. Yet, little to no validation was reported in each of the studies. If FE models are to address future research questions and clinical applications, then sound verification and validation of these models is required to ensure accurate conclusions and prediction of patient outcomes. Recommendations and considerations for future FE studies are therefore proposed.


Electrospun Sesbania Gum-Based Polymeric N-Halamines for Antibacterial Applications.
Microorganism pollution induced by pathogens has become a serious concern in recent years. In response, research on antibacterial N-halamines has made impressive progress in developing ways to combat this pollution. While synthetic polymer-based N-halamines have been widely developed and in some cases even commercialized, N-halamines based on naturally occurring polymers remain underexplored. In this contribution, we report for the first time on a strategy for developing sesbania gum (SG)-based polymeric N-halamines by a four-step approach Using SG as the initial polymer, we obtained SG-based polymeric N-halamines (abbreviated as cSG-PAN nanofibers) via a step-by-step controllable synthesis process. With the assistance of advanced techniques, the as-synthesized cSG-PAN nanofibers were systematically characterized in terms of their chemical composition and morphology. In a series of antibacterial and cytotoxicity evaluations, the as-obtained cSG-PAN nanofibers displayed good antibacterial activity against Escherichia coli and Staphylococcus aureus, as well as low cytotoxicity towards A549 cells. We believe this study offers a guide for developing naturally occurring polymer-based antibacterial N-halamines that have great potential for antibacterial applications.


[Efficacy and safety of extended-release clarithromycin (5-day short-course) vs telithromycin, in acute bacterial exacerbation of chronic bronchitis].
The extended-release formulation of clarithromycin (CLA-ER) allows using this macrolide as a single daily dose. The purpose of this study was to evaluate the efficacy and safety of the CLA-ER formulation (500 mgx2) vs telithromycin (TELI) (400 mgx2) as a short course 5-day treatment, once a day, in patients with AECB. This randomized double-blind study was conducted in patients with AECB without severe airflow limitation (FEV1>35%), with sputum purulence (mandatory criterion), and with either increased sputum volume or increased dyspnea, or both (Anthonisen criteria I or II). Three hundred sixty-two patients were assessed (62.6 years of age+/-12.9, men: 58.8%) positive culture on inclusion for 53.8%, with Haemophilus influenzae (N=57), Moraxella catarrhalis (N=42), and Streptococcus pneumoniae (N=41). In the per protocol population, the clinical success rate at day 8 was 97% (161/166) vs 97% (146/151), 97.5% CI=[-4.12 -4.71], the clinical cure rate at day 30 was 78% (129/166) versus 77% (116/151), P=0.85, and mean time without recurrence was 62 days versus 61 days (P=0.51), in CLA-ER and TELI groups, respectively. Fourteen patients in the CLA-ER group (8.2%) and 20 patients in the TELI group (12.4%) experienced at least one treatment-related adverse event (P=0.21), upon which gastrointestinal events were the most commonly reported treatment-related ones. CLA-ER (1000 mg once a day) for 5 days is at least as effective as telithromycin in the treatment of AECB without severe airflow limitation and is well tolerated.


Medial olivocochlear suppression in musicians versus non-musicians.
The medial olivocochlear efferent (MOCE) branch synapses with outer hair cells (OHCs), and the efferent pathway can be activated via a contralateral acoustic stimulus (CAS). The activation of MOCE can change OHC motile responses and convert signals that are capable of controlling the sensitivity of the peripheral hearing system in a frequency-specific manner. The aim of this study was to examine the MOCE system activity in professional musicians using transient evoked otoacoustic emission test and CAS. Musician group showed stronger suppression in all frequency bands in the presence of CAS.


Two-loop divergences of QCD scattering amplitudes with massive partons.
We provide a complete description of the two-loop infrared singularities of scattering amplitudes with an arbitrary number of massive and massless partons in non-Abelian gauge theories. To this end, we calculate the two universal functions that completely specify the structure of three-parton correlations in the soft anomalous-dimension matrix at two-loop order in closed analytic form. Both functions are found to be suppressed like O(m(4)/s(2)) in the limit of small parton masses, in accordance with mass factorization theorems. On the other hand, they are unsuppressed near the threshold for pair production of two heavy particles. As an application, we calculate the two-loop anomalous-dimension matrix for qq --> tt near threshold and show that it is not diagonal in the s-channel singlet-octet basis.


Multisensory perception and attention in school-age children.
Although it is well known that attention can modulate multisensory processes in adults and infants, this relationship has not been investigated in school-age children. Attention abilities of 53 children (ages 7-13 years) were assessed using three subscales of the Test of Everyday Attention for Children (TEA-Ch): visuospatial attention (Sky Search [SS]), auditory sustained attention (Score), and audiovisual dual task (SSDT, where the SS and Score tasks are performed simultaneously). Multisensory processes were assessed using the McGurk effect (a verbal illusion where speech perception is altered by vision) and the Stream-Bounce (SB) effect (a nonverbal illusion where visual perception is altered by sound). The likelihood of perceiving both multisensory illusions tended to increase with age. The McGurk effect was significantly more pronounced in children who scored high on the audiovisual dual attention index (SSDT). In contrast, the SB effect was more pronounced in children with higher sustained auditory attention abilities as assessed by the Score index. These relationships between attention and the multisensory illusory percepts could not be explained solely by age or children's intellectual abilities. This study suggests that the interplay between attention and multisensory processing depends on both the nature of the multisensory task and the type of attention needed to effectively merge information across the senses.


Recommended screenings for chlamydia and gonorrhea: A Canadian guidelines review.
Chlamydia and gonorrhea are the most commonly diagnosed bacterial sexually transmitted infections (STIs), and both are increasing in incidence. Because these STIs are often asymptomatic and found at extragenital sites, such as the pharynx and rectum, it is important that clinicians know when and how to screen for them. This article reviews recent updates to Canadian screening guidelines, including the evidence that led to updates on this topic.


An inexpensive, simple protocol for DNA isolation from blood for high-throughput genotyping by polymerase chain reaction or restriction endonuclease digestion.
We describe simple, inexpensive, and reliable methods for isolating DNA from avian blood, semen, or feather pulp. The procedures are readily applicable to high-throughput 96-well plate isolation for genotype analysis of chicken DNA based on restriction endonuclease digestion or PCR. Isolation cost is primarily the cost of a deep-well assay block and a few pipet tips; current price is less than 0.10 dollar per sample, providing a significant cost advantage over commercial kits. The procedure employs inexpensive, nonhazardous reagents and yields intact, double-stranded DNA from as little as 2 to 10 microL of avian blood, suitable for RFLP analysis or hundreds of PCR amplifications. We compared our method to published procedures for alkaline extraction from feather pulp and found our method to be more reliable with the advantage of isolating intact DNA sequences that can be easily quantified. With minor modifications, the method can isolate DNA for PCR genotyping from mammalian whole blood.


Ageism among social work faculty: impact of personal factors and other "isms".
The purpose of this article was (a) to determine the extent to which ageist attitudes are evident among social work faculty and how educational factors may contribute to ageism, (b) to determine if terror management theory (in terms of aging anxiety) offers a further explanation for ageist attitudes beyond known correlates, and (c) to understand how intersecting prejudices (attitudes toward women, gay men, and lesbians) may be associated with ageist attitudes. Results indicated a low bias toward older adults, with two variables, psychological anxiety about aging and paid experience with older adults, accounting for 29.7% of the variance. Further, no association was found between ageism and sexism and sexual prejudice in the multivariate analyses. These results indicate promising advances for terror management theory in explaining ageism. Social work faculty's low bias and perceived need for gerontological content in curricula is an encouraging finding for gerontological social work education.


So what shall we do about assertive community treatment?
The usefulness of Assertive Community Treatment (ACT) in European countries with well-developed community care systems has been disputed, despite considerable relevant literature. This paper aims to assess reasons for and against implementing ACT in such countries. ACT may not be useful where generic community mental health teams are not yet well-developed, where admission rates are already low, or where an alternative model based on close integration of a full range of types of care is in place. Good reasons for introducing ACT include listening to patients' preferences, being able to monitor a high risk group of patients more successfully, good staff satisfaction, and the potential for using ACT teams as a platform for delivering interventions for difficult to treat psychosis. The ACT model is more likely to thrive in future if a recovery orientation can be adopted.


Changing concepts in the management of pathological and impending pathological fractures.
The benefits in the management of pathological fractures by internal fixation are well recognized. However, failures in bone or metal can occur where there is a large amount of bone destruction. Seven of 69 pathological fractures in our series were treated by internal fixation with adjunctive use of methylmethacrylate without failure. This allowed for secure fixation in fractures with large amounts of bone destruction, thus decreasing the patient's pain and allowing earlier mobilization. Neither the effects of radiotherapy on bone involved with neoplasm nor methylmethacrylate are altered when radiotherapy is used after internal fixation and adjunctive use of methylmethacrylate. We believe that an aggressive program of stabilization of these fractures or fixation before actual fracture provides significant patient benefits including reduction of pain, decreased length of hospital stay, reduction of financial expense, and return to as near normal function as possible.


A comparative analysis of short stays versus long stays in adult day health care programs.
The purpose of this study is to identify challenges to continual use of adult day services. Individuals who stayed in five adult day health care programs for four weeks or less are compared to those who stayed enrolled more than four weeks. Data were gathered from new intake files recorded January 1, 1999 to December 31, 2000 at five adult day programs operating in central Ohio. This study is based on 143 clients who disenrolled during this study period. Short stay participants were significantly different from long stay participants in race, educational level, mental health, and in financial and social resources. The findings support the important partnerships between informal and formal care resources in the utilization of adult day services and ultimately community-based living.


The anti-HIV drug nelfinavir mesylate (Viracept) is a potent inhibitor of cell fusion caused by the SARSCoV-2 spike (S) glycoprotein warranting further evaluation as an antiviral against COVID-19 infections.
Severe acute respiratory syndrome coronavirus-2 (SARS CoV-2) is the causative agent of the coronavirus disease-2019 (COVID-19) pandemic. Coronaviruses enter cells via fusion of the viral envelope with the plasma membrane and/or via fusion of the viral envelope with endosomal membranes after virion endocytosis. The spike (S) glycoprotein is a major determinant of virus infectivity. Herein, we show that the transient expression of the SARS CoV-2 S glycoprotein in Vero cells caused extensive cell fusion (formation of syncytia) in comparison to limited cell fusion caused by the SARS S glycoprotein. Both S glycoproteins were detected intracellularly and on transfected Vero cell surfaces. These results are in agreement with published pathology observations of extensive syncytia formation in lung tissues of patients with COVID-19. These results suggest that SARS CoV-2 is able to spread from cell-to-cell much more efficiently than SARS effectively avoiding extracellular neutralizing antibodies. A systematic screening of several drugs including cardiac glycosides and kinase inhibitors and inhibitors of human immunodeficiency virus (HIV) entry revealed that only the FDA-approved HIV protease inhibitor, nelfinavir mesylate (Viracept) drastically inhibited S-n- and S-o-mediated cell fusion with complete inhibition at a 10-μM concentration. In-silico docking experiments suggested the possibility that nelfinavir may bind inside the S trimer structure, proximal to the S2 amino terminus directly inhibiting S-n- and S-o-mediated membrane fusion. Also, it is possible that nelfinavir may act to inhibit S proteolytic processing within cells. These results warrant further investigations of the potential of nelfinavir mesylate to inhibit virus spread at early times after SARS CoV-2 symptoms appear.


Three new cases of spondylocarpotarsal synostosis syndrome: clinical and radiographic studies.
Spondylocarpotarsal synostosis syndrome (SSS) or congenital synspondylism is a recently delineated clinical entity. At least 15 patients have been reported. We present 3 new patients, 2 of whom were sibs born to first-cousin parents. All of our patients had multiple synostoses involving cervical, thoracic and/or lumbar vertebral bodies and carpal/tarsal bones, scoliosis/lordosis, and short stature. Sensorineural deafness was found in 2 of the 3 patients. Analysis of clinical manifestations suggests clinical variability and genetic heterogeneity in SSS. Of a total of 18 SSS patients, 10 were five pairs of sibs from five families, with first-cousin consanguinity of parents in 3, indicating that at least one type of SS is an autosomal-recessive disorder.


Discovery and characterization of BlsE, a radical S-adenosyl-L-methionine decarboxylase involved in the blasticidin S biosynthetic pathway.
BlsE, a predicted radical S-adenosyl-L-methionine (SAM) protein, was anaerobically purified and reconstituted in vitro to study its function in the blasticidin S biosynthetic pathway. The putative role of BlsE was elucidated based on bioinformatics analysis, genetic inactivation and biochemical characterization. Biochemical results showed that BlsE is a SAM-dependent radical enzyme that utilizes cytosylglucuronic acid, the accumulated intermediate metabolite in blsE mutant, as substrate and catalyzes decarboxylation at the C5 position of the glucoside residue to yield cytosylarabinopyranose. Additionally, we report the purification and reconstitution of BlsE, characterization of its [4Fe-4S] cluster using UV-vis and electron paramagnetic resonance (EPR) spectroscopic analysis, and investigation of the ability of flavodoxin (Fld), flavodoxin reductase (Fpr) and NADPH to reduce the [4Fe-4S](2+) cluster. Mutagenesis studies demonstrated that Cys31, Cys35, Cys38 in the C×××C×MC motif and Gly73, Gly74, Glu75, Pro76 in the GGEP motif were crucial amino acids for BlsE activity while mutation of Met37 had little effect on its function. Our results indicate that BlsE represents a typical [4Fe-4S]-containing radical SAM enzyme and it catalyzes decarboxylation in blasticidin S biosynthesis.


Quantification of xylooligomers in hot water wood extract by 1H-13C heteronuclear single quantum coherence NMR.
A new method that employs 2D-HSQCNMRwasdeveloped to determine xylooligomer concentrations in the hot water extracts of Paulownia elongata, aspen, sugar maple, southern hardwood mixture, and willow woodchips. Equations for computing oligomer concentrations calculation were developed based on HSQC corresponding resonance integrals of xylooligomer C1H1 and monomeric sugar standard curves. The degree of polymerization (DP) of xylooligomers in the hot water extract was computed by equation obtained from a series of xylooligomer standard solutions with DPs that ranged from 2 to 6. Another group of hot water wood extract that is served as a control group was hydrolyzed by 4% sulfuric acid at 121 °C for 60 min in order to convert all xylooligomer into xylose. As 2D-HSQC resonance response is different for acetylated xylo-units, as compared with non-acetylated units, proton NMR was used to calibrate the acetylated xylooligomer concentration. Xylooligomer concentrations determined from HSQC compared fairly well with data after hydrolysis.


Value choices and considerations when limiting intensive care treatment: a qualitative study.
To shed light on the values and considerations that affect the decision-making processes and the decisions to limit intensive care treatment. Qualitative methodology with participant observation and in-depth interviews, with an emphasis on eliciting the underlying rationale of the clinicians' actions and choices when limiting treatment. Informants perceived over-treatment in intensive care medicine as a dilemma. One explanation was that the decision-making base was somewhat uncertain, complex and difficult. The informants claimed that those responsible for taking decisions from the admitting ward prolonged futile treatment because they may bear guilt or responsibility for something that had gone wrong during the course of treatment. The assessments of the patient's situation made by physicians from the admitting ward were often more organ-oriented and the expectations were less realistic than those of clinicians in the intensive care unit who frequently had a more balanced and overall perspective. Aspects such as the personality and the speciality of those involved, the culture of the unit and the degree of interdisciplinary cooperation were important issues in the decision-making processes. Under-communicated considerations jeopardise the principle of equal treatment. If intensive care patients are to be ensured equal treatment, strategies for interdisciplinary, transparent and appropriate decision-making processes must be developed in which open and hidden values are rendered visible, power structures disclosed, employees respected and the various perspectives of the treatment given their legitimate place.


A multicenter study of biopsied oral and maxillofacial lesions in a Brazilian pediatric population.
The aim of this study was to investigate the prevalence of oral and maxillofacial lesions among children from representative regions of Brazil. A retrospective descriptive study was conducted. Biopsy records comprising the period from 2000 to 2015 were obtained from the archives of three Brazilian oral pathology referral centers. A total of 32,506 biopsy specimens were analyzed, and specimens from 1,706 children aged 0-12 years were selected. Gender, age, anatomical location and histopathological diagnosis were evaluated. Descriptive statistics was carried out. Likelihood ratio tests were used to evaluate the association between the categorical variables. The level of significance was set at 0.05. The post-hoc test was used to identify the subgroups that significantly differed from one another, and the Bonferroni correction was applied. A total of 1,706 oral and maxillofacial lesions were diagnosed in pediatric patients, including 51.9% girls. Oral mucocele was the most prevalent reactive/inflammatory lesion (64%). The most commonly affected sites were the lips (34.5%) and mandible (19.9%). A significant association was observed between age and the group of lesions of the oral cavity (p < 0.001), and between age and anatomical location (p < 0.001). Pediatric oral and maxillofacial lesions were frequent and showed wide diversity, with the prevalence of mucocele. Knowledge of oral lesions is important for pediatric dentists worldwide, since it provides accurate data for the diagnosis and oral health of children.


Epinephrine-induced QT interval prolongation: a gene-specific paradoxical response in congenital long QT syndrome.
To determine the effect of epinephrine on the QT interval in patients with genotyped long QT syndrome (LQTS). Between May 1999 and April 2001, 37 patients (24 females) with genotyped LQTS (19 LQT1, 15 LQT2, 3 LQT3, mean age, 27 years; range, 10-53 years) from 21 different kindreds and 27 (16 females) controls (mean age, 31 years; range, 13-45 years) were studied at baseline and during gradually increasing doses of intravenous epinephrine infusion (0.05, 0.1, 0.2, and 0.3 microg x k(-1) x min(-1)). The 12-lead electrocardiogram was monitored continuously, and heart rate, QT, and corrected QT interval (QTc) were measured during each study stage. There was no significant difference in resting heart rate or chronotropic response to epinephrine between LQTS patients and controls. The mean +/- SD baseline QTc was greater in LQTS patients (500+/-68 ms) than in controls (436+/-19 ms, P<.001). However, 9 (47%) of 19 KVLQT1-genotyped LQT1 patients had a nondiagnostic resting QTc (<460 milliseconds), whereas 11 (41%) of 27 controls had a resting QTc higher than 440 milliseconds. During epinephrine infusion, every LQT1 patient manifested prolongation of the QT interval (paradoxical response), whereas healthy controls and patients with either LQT2 or LQT3 tended to have shortened QT intervals (P<.001). The maximum mean +/- SD change in QT (AQT [epinephrine QT minus baseline QT]) was -5+/-47 ms (controls), +94+/-31 ms (LQT1), and -87+/-67 ms (LQT2 and LQT3 patients). Of 27 controls, 6 had lengthening of their QT intervals (AQT >30 milliseconds) during high-dose epinephrine. Low-dose epinephrine (0.05 microg x kg(-1) x min(-1)) completely discriminated LQT1 patients (AQT, +82+/-34 ms) from controls (AQT, -7+/-13 ms; P<.001). Epinephrine-triggered nonsustained ventricular tachycardia occurred in 2 patients with LQTS and in 1 control. Epinephrine-induced prolongation of the QT interval appears pathognomonic for LQT1. Low-dose epinephrine infusion distinguishes controls from patients with concealed LQT1 manifesting an equivocal QTc at rest. Thus, epinephrine provocation may help unmask some patients with concealed LQTS and strategically direct molecular genetic testing.


Enzyme-assisted nanolithography.
We have chemically immobilized alkaline phosphatase molecules onto the apex of a tip of an atomic force microscope. When the substrate BCIP is dephosphorylated by alkaline phosphatase, it will precipitate in the presence of NBT. By bringing the tip in the vicinity of a suitable sample, we could locally deposit this complex on the sample. Thus we combined the activity of an enzyme with the accuracy in positioning a tip in scanning probe microscopy to demonstrate a novel technique referred to as enzyme-assisted nanolithography. By use of other enzymes, this method will open the possibility to chemically modify surfaces on a nanometer scale.


Weight gain among men and women who have a child enter their home.
Parenthood is associated with decreased physical activity and dietary changes. Previously, mothers have been the focus of studies examining the influence of children on parents' body mass index (BMI), largely ignoring whether parenting affects fathers. This study assessed weight gain in mothers and fathers (by birth or other), using longitudinal repeated-measures models to assess BMI changes over time; parents were compared with nonparents. Data were from the Coronary Artery Risk Development in Young Adults cohort study and included 2,881 black and white adults, ages 18 to 30 years, without children at baseline (1985-1986), and from four urban locations. At each time point (years 2, 5, and 7), changes in BMI from baseline were analyzed, comparing those who had their children in their household at that time point (parents) and those without children (nonparents). The "child effect" is the mean difference in BMI change in parents compared with nonparents. In fathers, overall, the child effect was not significant (black males: 0.30; P=0.09; white males: 0.03; P=0.77). Among black men, however, interactions between age and parental status were significant (P=0.02). Black men who were aged 18 to 24 years at baseline and became fathers during the next 7 years demonstrated a significant child effect, gaining an average of 0.68 more in BMI than nonfathers (P=0.003). Mothers of both races demonstrated the child effect; for blacks it was 0.65 (P=0.003) and for whites it was 1.12 (P≤0.001). These data reveal that becoming a parent can affect the BMI of some adults and suggest that obesity-prevention interventions for children and adult-focused healthy-lifestyle interventions could have additional impact through a family focus targeting both parent and child outcomes.


Spectrum of biopsy proven renal diseases in Central China: a 10-year retrospective study based on 34,630 cases.
Chronic kidney diseases have become a major issue worldwide. The spectrum of biopsy proven renal diseases differs between locations and changes over time. It is therefore essential to describe the local epidemiological trends and the prevalence of renal biopsy in various regions to shine new light on the pathogenesis of various renal diseases and provide a basis for further hypothesis-driven research. We retrospectively analyzed 34,630 hospitalized patients undergoing native renal biopsy between January 1, 2009 and December 31, 2018. Indications for renal biopsy and histological diagnosis were analyzed to describe the prevalence of renal biopsy, and changing prevalence between period 1 (2009-2013) and period 2 (2014-2018) were further analyzed. Nephrotic syndrome (NS) was the most common indication for biopsy. Membranous nephropathy (MN, 24.96%) and IgA nephropathy (IgAN, 24.09%) were the most common primary glomerulonephritis (PGN). MN was most common in adults, with IgAN more prevalent in children. Lupus nephritis (LN) was the most common secondary glomerulonephritis (SGN) in adults, while Henöch-Schönlein purpura nephritis (HSPN) in children. The prevalence of MN increased significantly and nearly doubled from period 1 (15.98%) to period 2 (30.81%) (P = 0.0004). The same trend appeared with membranoproliferative glomerulonephritis (MPGN), diabetic nephropathy (DN) and obesity-related glomerulopathy (ORG), while the frequencies of minimal change disease (MCD), focal segmental glomerulosclerosis (FSGS), LN and hepatitis B associated glomerulonephritis (HBV-GN) significantly decreased between the two intervals. NS was the most common indication for biopsy across all age groups and genders. MN has overtaken IgAN to become the most common PGN in adults, while IgAN was the most common PGN in children. LN was the most common SGN in adults, and HSPN the most common in children.


Metasurfaces with high-Q resonances governed by topological edge state.
Achieving high-quality (Q)-factor resonances in metasurfaces is essential for various applications, including nano-lasers, nonlinear optics, and quantum optics. In this work, we propose a high-Q metasurface using a topological strategy: constructing the metasurface by stacking two conjugated nanopillar arrays with different topological invariants. Our study shows that a topological edge state steadily appears at the interfaces of the nanopillars, and a sharp transmission resonance with a Q-factor of more than 1000 can be obtained. The sensing application of such high-Q topological metasurface is also demonstrated, whose figure of merit reaches approximately 145. The proposed strategy and underlying theory can open up new avenues to realize ultrasharp resonances, which can promote numerous potential applications, such as biosensing, optical modulation, and slow-light devices.


Chirality-driven mass enhancement in the kagome Hubbard model.
We investigate the quasiparticle-mass enhancement in the Hubbard model on the frustrated kagome lattice by using a cluster extension of the dynamical mean-field theory. By analyzing the cluster density matrix, we find a hierarchy of energy scale among charge, spin, and chirality degrees of freedom. A large amount of entropy associated with the chirality is released at a much lower temperature than other energy scales for spin and charge fluctuations, leading to a sharp peak in the specific heat and the single-particle spectrum. The results manifest a generic mechanism of mass enhancement driven by an emergent composite degree of freedom under geometrical frustration.


The Pro-Oncogenic Sphingolipid-Metabolizing Enzyme β-Galactosylceramidase Modulates the Proteomic Landscape in BRAF(V600E)-Mutated Human Melanoma Cells.
β-Galactosylceramidase (GALC) is a lysosomal enzyme involved in sphingolipid metabolism by removing β-galactosyl moieties from β-galactosylceramide and β-galactosylsphingosine. Previous observations have shown that GALC may exert pro-oncogenic functions in melanoma and Galc silencing, leading to decreased oncogenic activity in murine B16 melanoma cells. The tumor-driving BRAF(V600E) mutation is present in approximately 50% of human melanomas and represents a major therapeutic target. However, such mutation is missing in melanoma B16 cells. Thus, to assess the impact of GALC in human melanoma in a more relevant BRAF-mutated background, we investigated the effect of GALC overexpression on the proteomic landscape of A2058 and A375 human melanoma cells harboring the BRAF(V600E) mutation. The results obtained by liquid chromatography-tandem mass spectrometry (LC-MS/MS) demonstrate that significant differences exist in the protein landscape expressed under identical cell culture conditions by A2058 and A375 human melanoma cells, both harboring the same BRAF(V600E)-activating mutation. GALC overexpression resulted in a stronger impact on the proteomic profile of A375 cells when compared to A2058 cells (261 upregulated and 184 downregulated proteins versus 36 and 14 proteins for the two cell types, respectively). Among them, 25 proteins appeared to be upregulated in both A2058-upGALC and A375-upGALC cells, whereas two proteins were significantly downregulated in both GALC-overexpressing cell types. These proteins appear to be involved in melanoma biology, tumor invasion and metastatic dissemination, tumor immune escape, mitochondrial antioxidant activity, endoplasmic reticulum stress responses, autophagy, and/or apoptosis. Notably, analysis of the expression of the corresponding genes in human skin cutaneous melanoma samples (TCGA, Firehose Legacy) using the cBioPortal for Cancer Genomics platform demonstrated a positive correlation between GALC expression and the expression levels of 14 out of the 27 genes investigated, thus supporting the proteomic findings. Overall, these data indicate for the first time that the expression of the lysosomal sphingolipid-metabolizing enzyme GALC may exert a pro-oncogenic impact on the proteomic landscape in BRAF-mutated human melanoma.


Promoting social capital to alleviate loneliness and improve health among older people in Spain.
Loneliness is especially frequent among older people in Southern Europe. Furthermore, promoting social capital to tackle loneliness and its health effects is an understudied intervention strategy. Therefore, a complex intervention was piloted in Spain in a pre-post study with a 2-year follow-up. Its aims were to explore the feasibility of the intervention and its short- and long-term effects. It was conducted in one mixed rural-urban and two urban areas of diverse socioeconomic levels from 2011 to 2012. The intervention framework was based on social capital theory applying a behaviour change model and care co-ordination. The intervention comprised: (i) a co-ordinated action aimed at building a network between primary healthcare centres and community assets in the neighbourhood and (ii) a group-based programme, which promoted social capital among lonely older people, especially social support and participation. Older people active in senior centres volunteered as gatekeepers. The main outcome domain was loneliness. Secondary outcome domains were participation, social support, self-perceived health, quality of life, depressive symptoms and use of health resources. Pre-post changes were assessed with t-test, Wilcoxon signed-rank test and McNemar's test. Differences between the three time points were assessed with a one-way ANOVA with repeated measures. Social workers and nurses were successfully involved as group leaders, 10 volunteers took part and 38 participants were included. After the intervention, loneliness decreased while social participation and support significantly increased. Furthermore, the number of visits to nurses increased. Exactly 65.8% of the participants built social contacts within the group and 47.4% became engaged in new activities. Two years later, social effects were maintained and depressive symptoms had decreased. Exactly 44.7% of the participants continued to be in contact with at least one person from the group and 39.5% continued participating. The intervention contributes a novel and feasible social capital-based approach for alleviating loneliness among older adults while prompting meaningful changes in their lives.


Physicochemical characterizations of piroxicam-poloxamer solid dispersion.
To develop an efficient topical delivery system for piroxicam using poloxamer gel formulation, physicochemical behavior of piroxicam in poloxamer was studied. The gelling property of poloxamer and the solubility of piroxicam in the poloxamer were investigated. The interaction between piroxicam and poloxamer was studied by x-ray diffractometry (XRD), infrared (IR) spectroscopy and differential thermal analysis (DTA) with a solid dispersion, coprecipitate, or physical mixture. Poloxamer 407 solutions showed the property of a gel when the concentration was higher than 15% (w/w) and poloxamer 407 increased the aqueous solubility of piroxicam by about 11-fold at the concentration of 22.5% (w/w). The results of XRD did not show the crystalline from of piroxicam in the solid dispersion and results of IR spectroscopic analysis showed an association between functional groups of piroxicam and poloxamer.


Stereologic studies on mitochondrial configuration in mitochondria of immobilized mice muscle.
The effect of long-term (3 weeks) immobilization on mice skeletal (m. gastrocnemius) and heart muscles was investigated. Morphometric determinations were carried out for analysis surface and volume parameters in mitochondria. The ultrastructural visual evaluation of mitochondria showed that there are no difference between control and experimental groups. Morphometric data obtained from stereological analysis were shown that mitochondria in immobilized skeletal and heart muscles represent tendency of transformation to low-energy states (condensed state). Based upon data obtained in this study we recommend that these changes in mitochondria there are reversible.


[Biosynthesis of sterols by various yeasts].
Biosynthesis of sterines by yeasts was studied with 112 strains belonging to 9 genera; 77 strains synthesized ergosterine and 15 most active strains were selected, which belonged to the following genera: Saccharomyces, Torulopsis, Hanseniaspora, Debaryomyces, Candida, Mechnikowia. Sterine composition was studied in these active strains. The main sterine in all studied strains was ergosterine.


A novel thiolase-reductase gene fusion promotes the production of polyhydroxybutyrate in Arabidopsis.
The production of polyhydroxybutyrate (PHB) involves a multigene pathway consisting of thiolase, reductase and synthase genes. In order to simplify this pathway for plant-based expression, a library of thiolase and reductase gene fusions was generated by randomly ligating a short core linker DNA sequence to create in-frame fusions between the thiolase and reductase genes. The resulting fusion constructs were screened for PHB formation in Escherichia coli. This screen identified a polymer-producing candidate in which the thiolase and reductase genes were fused via a 26-amino-acid linker. This gene fusion, designated phaA-phaB, represents an active gene fusion of two homotetrameric enzymes. Expression of phaA-phaB in E. coli and Arabidopsis yielded a fusion protein observed to be the expected size by Western blotting techniques. The fusion protein exhibited thiolase and reductase enzyme activities in crude extracts of recombinant E. coli that were three-fold and nine-fold less than those of the individually expressed thiolase and reductase enzymes, respectively. When targeted to the plastid, and coexpressed with a plastid-targeted polyhydroxyalkanoate (PHA) synthase, the fusion protein enabled PHB formation in Arabidopsis, yielding roughly half the PHB formed in plants expressing individual thiolase, reductase and synthase enzymes. This work represents a first step towards simplifying the expression of the PHB biosynthetic pathway in plants.


[Prospects of the development of quality control technologies for traditional Chinese medicine].
Quality control is one of bottleneck problem limiting the applications and development of traditional Chinese medicines (TCMs). In recent years, the research on TCMs has already made a great progress. In this review, the background of quality control is discussed from the requirement of TCMs industry, modernization of TCMs, the requirement of techniques as well as opportunities and challenges in the quality control of TCMs. The significance of quality control to improve the efficiency and safety of TCMs, to promote the development of TCMs industry and the globalization of TCMs are also discussed. The status quo of quality control is reviewed, in which the drawbacks of process control, safety control, the preparation of reference compounds and fingerprint in quality control of TCMs were analyzed. In addition, the technological prospects based on modern separation techniques and detection methods for the quality control of TCMs are put forward, including the key techniques of the quality control, the techniques of safety control, the standard systems of the quality control, the innovative techniques, the techniques for the preparation of reference compounds, the establishment of technique standards, etc.


Obesity-induced upregulation of myocardial endothelin-1 expression is mediated by leptin.
Several studies have shown that leptin, the product of the obese gene, may link obesity with cardiovascular diseases, and in particular with cardiac hypertrophy. In vitro studies suggest that the mechanism by which leptin causes cardiac hypertrophy involves the upregulation of endogenous endothelin-1 (ET-1), a potent vasoconstrictor and mitogen. Whether obesity-associated hyperleptinemia causes an increase in myocardial ET-1 expression in vivo remains unclear. To address this issue, we fed mice with a high-fat diet and analyzed serum levels of ET-1 and ET-1 mRNA in the heart. We found that in mice fed a high-fat diet, serum ET-1, myocardial ET-1, leptin and leptin receptor mRNA were all elevated. In contrast, in leptin-deficient obese (ob/ob) mice, both serum and myocardial ET-1 levels were not higher than in wild type mice. These findings suggest that upregulation of myocardial ET-1 by obesity is mediated by leptin.


Correcting the use of ensemble averages in the calculation of harmonics to noise ratios in voice signals (L).
A correcting formula for the estimation of harmonics-to-noise ratios (HNR) based on ensemble-averaging techniques is derived. The original method yields a biased approximation which is more accurate as the number of averaged pulses (N) increases. However, the method treats gradual waveform changes incorrectly as noise, which is worsened for large values of N. The obtained formula allows the use of as few averaged pulses as desired, while allowing the complete removal of the bias from the estimate of HNR.


The Stockholm birth centre trial: maternal and infant outcome.
To compare an in-hospital birth centre with standard maternity care regarding medical interventions and maternal and infant outcome. The birth centre care was characterised by comprehensive antenatal, intrapartum and postpartum care with the same team of midwives, restricted use of medical technology, and discharge within 24 h after birth. Of 1860 women meeting low risk medical criteria in early pregnancy and interested in birth centre care, 928 were randomly allotted birth centre care and 932 standard maternity care. Data were collected mainly from hospital records, and analysis was by intention-to-treat. Of the women in the birth centre group, 13% were transferred antenatally and 19% intrapartum. No statistical differences were observed in maternal morbidity or in perinatal mortality, neonatal morbidity, Apgar score or infant admissions to neonatal care. Perinatal mortality, defined as intrauterine death after 22 weeks of gestation and infant death within seven days of birth, occurred in eight cases (0.9%) in the birth centre group and in two cases (0.2%) in the standard care group (OR 4.04, 95% CI 0.80 to 39.17; P = 0.11). Subgroup analysis showed that a larger proportion of first-born babies in the birth centre group (15.6%) were admitted for neonatal care than in the standard care group (9.5%) (P = 0.003), whereas the converse was the case for the newborns of multiparous women: 4.7% and 8.4%, respectively (P = 0.04). The overall rates of operative delivery (e.g. caesarean section, vacuum extraction and forceps), 11.1% in the birth centre group and 13.4% in the standard care group, did not differ statistically (P = 0.12), but obstetric analgesia, induction, augmentation of labour and electronic fetal monitoring were less frequently used in the birth centre group. Labour was 1 h longer in the birth centre group. Birth centre care was associated with less medical interventions than standard care without any statistically significant differences in health outcomes. However, the excess of perinatal deaths and of morbidity in primigravidas' infants in the birth centre group gives cause for concern and necessitates further studies.


Fibromuscular dysplasia as the substrate for systemic and pulmonary hypertension in the setting of Moya-Moya disease.
We report the first case, to the best of our knowledge, of a child with the unusual association of Moya-Moya disease and both systemic and pulmonary hypertension. Histological examination revealed fibromuscular dysplasia as the common denominator for a diffuse arteriopathy.


Compounds with the dioxopyrimidine cycle inhibit cholinesterases from different groups of animals.
We firstly synthesized derivatives of 6-methyluracil, alloxazine, and xanthine, containing omega-tetraalkylammonium (TAA) groups at the N(1) and N(3) atoms in a pyrimidine cycle and assayed their anticholinesterase activities. Compounds with triethylpentylammoniumalkyl groups behaved as typical reversible inhibitors of acetylcholinesterase (AChE) (pI(50) 3.20-6.22) and butyrylcholinesterase (BuChE) (pI(50) 3.05-5.71). Compounds, containing two ethyl residues and a substituted benzyl fragment in the tetraalkylammonium group at N(3) atoms or two similar TAA groups at N(1) and N(3) atoms, possessed very high anticholinesterase activity. Although these compounds displayed the activity of typical irreversible AChE inhibitors (a progressive AChE inactivation; k(i) 7.6 x 10(8) to 3.5 x 10(9)M(-1)min(-1)), they were reversible inhibitors of BuChE (pI(50) 3.9-6.9). The efficiency of AChE inhibition by some of these compounds was more than 10(4) times higher than the efficiency of BuChE inhibition. Several synthesized TAA derivates of 6-methyluracil reversibly inhibited electric eel and cobra venom AChEs and horse serum BuChE. However, depending on their structure, the tested compounds possessed the time-progressing inhibition of mammalian erythrocyte AChE, typically of irreversible inhibitors. As shown upon dialysis and gel-filtration, the formed mammalian AChE-inhibitor complex was stable. Thus, a new class of highly active, selective, and irreversible inhibitors of mammalian AChE was described. In contrast to classical phosphorylating or carbamoylating AChE inhibitors, these compounds are devoid of acylating functions. Probably, these inhibitors interact with certain amino acid residues at the entrance to the active-site gorge.


Non-Eosinophilic Nasal Polyps Shows Increased Epithelial Proliferation and Localized Disease Pattern in the Early Stage.
Non-eosinophilic nasal polyps (NPs) show less inflammatory changes and are less commonly associated with lower airway inflammatory disorders such as asthma, compared with eosinophilic NPs. However, the development of non-eosinophilic NPs which is a predominant subtype in Asian population still remains unclear. A total of 81 patients (45 with non-eosinophilic NPs and 36 with eosinophilic NPs) were enrolled. Clinical information and computed tomography (CT), endoscopic, and histological findings were investigated. Tissue samples were analyzed for total IgE levels and for mRNA expression levels of interleukin (IL)-4, IL-5, IL-13, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, IL-17A, IL-22, IL-23p19, transforming growth factor (TGF)-β1, TGF-β2, TGF-β3, and periostin. Immunostaining assessment of Ki-67 as a proliferation marker was performed. We found that epithelial in-growing patterns such as pseudocysts were more frequently observed in histological and endoscopic evaluations of non-eosinophilic NPs, which was linked to increase epithelial staining of Ki-67, a proliferating marker. Eosinophilic NPs were characterized by high infiltration of inflammatory cells, compared with non-eosinophilic NPs. To investigate the developmental course of each subtype, CT was analyzed according to CT scores and subtypes. Non-eosinophilic NPs showed more localized pattern and maxillary sinus involvement, but lesser olfactory involvement in early stage whereas eosinophilic NPs were characterized by diffuse ethmoidal and olfactory involvement. In addition, high ethmoidal/maxillary (E/M) CT scores, indicating ethmoidal dominant involvement, were one of surrogate markers for eosinophilic NP. E/M CT scores was positively correlated with levels of TH2 inflammatory markers, including IL-4, IL-5, periostin mRNA expression and total IgE levels in NPs, whereas levels of the TH1 cytokine, IFN- γ were inversely correlated. Moreover, if the combinatorial algorithm meet the three of the four markers, including IL-5 (<2.379), periostin (<3.889), IFN-γ (>0.316), and E/M ratio (<2.167), non-eosinophilic CRSwNP are diagnosed with a sensitivity of 84.4% and a specificity of 84.8%. Histologic, immunologic and clinical data suggest that non-eosinophilic NPs showed enhanced epithelial alteration and more localized maxillary involvement. Combination of cutoff value on IL-5, periostin, IFN-γ, and E/M scores may be one of surrogate markers for non-eosinophil NP subtype.


Permeability of alkali metal cations in lobster muscle. A comparison of electrophysiological and osmometric analyses.
Single muscle fibers from lobster walking legs are effectively impermeable to Na, but are permeable to K. They shrink in hyperosmotic NaCl; they swell in low NaCl media which are hyposmotic or which are made isosmotic with the addition of KCl. In conformity, the membrane potential is relatively insensitive to changes in external Na, while it responds according to the Nernst relation for changes in external K. When the medium is made isosmotic or hyperosmotic with RbCl the volume and membrane potential changes are of essentially the same magnitudes as those in media enriched with KCl. The time courses for attaining equilibrium are slower, indicating that Rb is less permeant than K. Substitution of CsCl for NaCl (isosmotic condition) produces no change in volume of the muscle fiber. Addition of CsCl (hyperosmotic condition) causes a shrinkage which attains a steady state, as is the case in hyperosmotic NaCl. Osmotically, therefore, Cs appears to be no more permeant than is Na. However, the membrane depolarizes slowly in Cs-enriched media and eventually comes to behave as an ideal Cs electrode. Thus, the electrode properties of the lobster muscle fiber membrane may not depend upon the diffusional relations of the membrane and ions, and the osmotic permeability of the membrane for a given cation may not correspond with the electrophysiologically deduced permeability. Comparative data on the effects of NH(4) and Li are also included and indicate several other degrees of complexity in the cell membrane.


A case of dermatopathia pigmentosa reticularis with wiry scalp hair and digital fibromatosis resulting from a recurrent KRT14 mutation.
We report a patient of Malay ancestry with dermatopathia pigmentosa reticularis (DPR) resulting from a recurrent KRT14 p.R125C mutation. The patient has reticulate hyperpigmentation over his trunk and proximal limbs, together with onychodystrophy. Despite the absence of noncicatricial alopecia, he has acral nonscarring blisters, palmoplantar hyperkeratosis and hypoplastic dermatoglyphics, in addition to unusual abnormalities such as wiry scalp hair and digital fibromatous thickening.


Atomically precise placement of single dopants in si.
We demonstrate the controlled incorporation of P dopant atoms in Si(001), presenting a new path toward the creation of atomic-scale electronic devices. We present a detailed study of the interaction of PH3 with Si(001) and show that it is possible to thermally incorporate P atoms into Si(001) below the H-desorption temperature. Control over the precise spatial location at which P atoms are incorporated was achieved using STM H lithography. We demonstrate the positioning of single P atoms in Si with approximately 1 nm accuracy and the creation of nanometer wide lines of incorporated P atoms.


Bioethics for clinicians: 10. Research ethics.
Medical research involving human subjects raises complex ethical, legal and social issues. Investigators sometimes find that their obligations with respect to a research project come into conflict with their obligations to individual patients. The ethical conduct of research rests on 3 guiding principles: respect for persons, beneficience, and justice. Respect for persons underlies the duty to obtain informed consent from study participants. Beneficence demands a favourable balance between the potential benefits and harms of participation. Justice requires that vulnerable people not be exploited and that eligible candidates who may benefit from participation not be excluded without good cause. Studies must be designed in a way that ensures the validity of findings and must address questions of sufficient importance to justify the risks of participation. In any clinical trial there must be genuine uncertainty as to which treatment arm offers the most benefit, and placebo controls should not be used if effective standard therapies exist. Researchers have a responsibility to inform themselves about the ethical, legal and policy standards that govern their activities. When difficulties arise, they should consult the existing literature and seek the advice of experts in research ethics.


Five years' experience with levonorgestrel-releasing IUDs.
Two levonorgestrel-releasing IUDs and a copper-releasing IUD of the same shape were studied in a randomized comparative study over five years. The levonorgestrel-releasing IUDs released 20 micrograms or 30 micrograms per day. The Pearl index during the 10,600 woman-months of LNG-IUD use was 0.11. The control device releasing copper had a Pearl index of 1.6. The amount and duration of menstrual bleeding was greatly reduced, leading to a high incidence of oligo- or amenorrhea. The continuation rate in this pioneer trial was 53 per 100 users for the levonorgestrel-releasing IUD (LNG-IUDs) and 50 per 100 users for the copper-releasing IUD (Nova T). The removal rates for reasons other than amenorrhea were not significantly different. Discontinuation because of amenorrhea occurred during the first two years, the cumulative termination rate for this reason was 11.6 per 100 users at five years. The LNG-IUDs removed for investigation after five years of use revealed that the devices contained about 40 percent of the original load. The effective lifespan of the device has been demonstrated by this study to be five years; the residual steroid gives an additional safety period of two more years. The LNG-IUD is a highly effective reversible contraceptive method, which strongly reduced the amount and duration of bleeding. During the first two months there is scanty but frequent spotting which, like the high incidence of oligo- and/or amenorrhea, requires counselling of health personnel and women using LNG-IUDs.


[Evaluation of the sphincter function and rectal sensitivity in rectal prolapse associated with fecal incontinence].
It has been suggested that incontinent patients with rectal prolapse develop a relaxation of the internal sphincter at a lower filling volume than those with the same disorder who were continent. A constant relaxation of the internal sphincter during filling before the individual experiences a need to defecate could be a contributory cause of the incontinence in these patients. Aiming to evaluate the former observations we have investigated continent and incontinent patients, with complete and internal rectal prolapse, by mean of the recto-anal manovolumetry. 31 patients were studied, 21 had a complete rectal prolapse and 10 had internal rectal prolapse and 28 sex and age matched controls were selected among volunteers. No difference has been found between the continent and incontinent patients with rectal prolapse, in term of sensory function studied by graded isobaric distention. Moreover, no difference has been found between the rectal volume at which perception of filling and relaxation of the internal sphincter during filling occurs. Therefore, in this respect, the present data are not in agreement with the theory proposed.


Exploring pathogenesis of primary biliary cholangitis by proteomics: A pilot study.
To explore the pathogenesis of primary biliary cholangitis (PBC) by identifying candidate autoantibodies in serum samples by proteomics and bioinformatics. Nine antimitochondrial antibody (AMA)-positive PBC patients and nine age- and sex-matched AMA-negative PBC patients were recruited. Antigen enrichment technology was applied to capture autoantigens of human intrahepatic biliary epithelial cells (HiBECs) that are recognized by autoantibodies from the sera of PBC patients. Candidate autoantigens were identified by label-free mass spectrometry. Bioinformatics analysis with MaxQuant software (version 126.96.36.199), DAVID platform, and Cytoscape v.3.0 allowed illustration of pathways potentially involved in the pathogenesis of PBC. In total, 1081 candidate autoantigen proteins were identified from the PBC patient pool. Among them, 371 were determined to be significantly differentially expressed between AMA-positive and -negative PBC patients (P < 0.05). Fisher's exact test was performed for enrichment analysis of Gene Ontology protein annotations (biological processes, cellular components, and molecular functions) and the Kyoto Encyclopedia of Genes and Genomes pathways. Significantly different protein categories were revealed between AMA-positive and -negative PBC patients. As expected, autoantigens related to mitochondria were highly enriched in AMA-positive PBC patients. However, lower levels of AMA were also detected in AMA-negative PBC patients. In addition, autoantigens of AMA-negative PBC patients were mainly involved in B-cell activation, recognition of phagocytosis, and complement activation. AMA-negative PBC individuals may not exist, but rather, those patients exhibit pathogenesis pathways different from those of AMA-positive PBC. Comprehensive research is needed to confirm these observations.


Complete Genome Sequence of Veillonella atypica OK5, the First Transformable Strain in the Species.
The Veillonella atypica strain OK5 was isolated from a human saliva sample and was the first strain shown to be genetically transformable in the Veillonella genus. Genetic studies using this strain have helped us gain much insight into the ecology of human oral biofilms. Here, we report the complete genome sequence of V. atypica OK5.


Pharmacological management of nonalcoholic fatty liver disease in type 2 diabetes.
The prevalence of nonalcoholic fatty liver disease (NAFLD) in patients with type 2 diabetes (T2D) is high and it is associated with poor prognosis. Hepatic steatosis results as a consequence of excessive hepatic lipid accumulation which correlates with insulin resistance and lipotoxicity, with subsequent oxidative stress, inflammation, apoptosis and fibrosis. Areas covered: This article presents the main pathophysiologic mechanisms and currently available drugs evaluated for their therapeutic effects on NAFLD/nonalcoholic steatohepatitis (NASH) and drugs under development that target relevant pathogenetic pathways. However, to date there is no particular drug approved for treatment of NAFLD in patients with T2D. Expert commentary: Early recognition and intervention are essential to ameliorate disease progression. Specific recommendations are still needed for NAFLD/NASH screening and diagnosis and therapeutic algorithm in patients with T2D. Lifestyle optimization with significant weight loss is a key intervention in patients with NAFLD and T2D. Pioglitazone, liraglutide, vitamin E, OCA and pentoxifylline have proven some histological improvements in NASH and omega 3-PUFAs were shown to decrease liver fat, but no specific recommendation can be made for treatment of NASH. Perhaps a combination of agents that target different pathogenic pathways are needed to better control disease progression, but more robust evidence for these agents is still needed.


Sugar-coated signaling in follicular lymphoma.
In this issue of Blood, complementary studies by Amin et al and Linley et al demonstrate that sugar moieties linked to surface immunoglobulin (sIg) of follicular lymphoma (FL) cells directly interact with endogenous lectins within the lymphoma niche and lead to activation of downstream B-cell receptor (BCR) signaling pathways. In addition to providing further insight into the role of the microenvironment in lymphomagenesis, these findings expose a unique molecular interaction that may represent a viable target for therapeutic intervention.


A Hybrid Approach for Efficient Modeling of Medium-Frequency Propagation in Coal Mines.
An efficient procedure for modeling medium frequency (MF) communications in coal mines is introduced. In particular, a hybrid approach is formulated and demonstrated utilizing ideal transmission line equations to model MF propagation in combination with full-wave sections used for accurate simulation of local antenna-line coupling and other near-field effects. This work confirms that the hybrid method accurately models signal propagation from a source to a load for various system geometries and material compositions, while significantly reducing computation time. With such dramatic improvement to solution times, it becomes feasible to perform large-scale optimizations with the primary motivation of improving communications in coal mines both for daily operations and emergency response. Furthermore, it is demonstrated that the hybrid approach is suitable for modeling and optimizing large communication networks in coal mines that may otherwise be intractable to simulate using traditional full-wave techniques such as moment methods or finite-element analysis.


[Experimental study on the role of the mylohyoid muscle in the instability of dental prostheses].
Using cinematographic and radiocinematographic recording techniques, the authors studied the movements of inferior prosthesis unstable in the lingual region. Pronunciation of the diphthong La... La... La... causes upward and forward projection of the prosthesis. Electromyography records contraction of the mylo-hyoid during pronunciation of these diphthongs. In the totally edentulous patient, the crest of the mylo-hoid is very close to the alveolar crest. From a clinical standpoint, functional exploration of the mylohyoid is easy, the index finger being placed along the mylo-hyoid crest and the patient asked to say La... La... La... If the index finger is pushed forwards and up, surgical intervention to lower the mylo-hyoid (Trauner's operation) is indicated, in association or not with other procedures designed to treat the other causes of instability or the prosthesis.


ALK Inhibitors for Treating Cancer, Blood, and Kidney Diseases.
Cancers that traditionally have been resistant to apoptosis via chemical- or radiation-based therapies may respond when the treatments are combined with ALK-5 inhibition. Disclosures in this Patent Highlight provide inhibitors of activin-receptor-like kinases such as ALK-5, compositions and methods for increasing red blood cell or hemoglobin levels, and activin antagonists to treat, prevent, or reduce the progression rate or severity of kidney disease.


[An analysis of microorganisms isolated from blood samples of patients treated at the Department of Nephrology and Internal Diseases].
The development of invasive medical procedures involves a decrease of prevalence of generalized infections. Hemodialysed patients are at higher risk of infection. The aim of this study was to analyze microorganisms isolated from blood samples of patients treated at the Department of Nephrology and Internal Diseases of the Medical University in Bydgoszcz. The analysis was based on 959 blood samples obtained from 248 patients during the investigation period from 01.02.2001 to 30.11.2002. Among patients included in this analysis 52.0% were males, 48.0% females. The age oscillated between 18 and 95 years, average 58.6 +/- 17.6. Blood cultures were processed in the blood culture monitoring systems. Identification of isolated microorganisms was based on their morphological and biochemical features. Antibiotic sensitivity tests were conducted according to the recommendations of the NCCLS and National Reference Centre for Drug-Sensitivity of Microorganisms. Positive culture results were obtained for 151 (15.8%) samples. 1114 (75.5%) strains of Gram(+) bacteria and 36 (23.8%) strains of Gram(-) bacteria and one Candida albicans strain were isolated. The results of antibiotic sensitivity tests are also presented. High prevalence of CNS strains in our research may suggest contamination of the samples with normal skin flora, since those strains were usually only isolated one time. It might also be the result of the profile of the Department. The prevention of vascular access-related septicaemia is a vital element of the care of the hemodialysed patient, as it is a well-established risk factor of generalized infections. Elaborating of local standards for empirical antibiotic therapy of generalized infections based on the epidemiological data of a particular department or hospital ward is necessary.


Determination of residual moisture in lyophilized protein pharmaceuticals using a rapid and non-invasive method: near infrared spectroscopy.
This study examined the application of near-infrared (NIR) spectroscopy to analyze residual moisture in lyophilized protein pharmaceuticals sealed in glass vials. We demonstrated that NIR was able to determine residual moisture in five marketed and clinical products with the same precision as Karl Fischer titration. We further investigated how changes in product configuration and protein formulation affected NIR measurement accuracy using a lyophilized monoclonal antibody rhuMAb E25 containing 1% to 5% residual moisture. The results indicated that the lyophilized cake porosity and dimensions had no effect on NIR measurement when the cake height and diameter exceeded the NIR penetration depth. In addition, changing the buffer and surfactant concentrations in the formulation did not affect moisture determination by NIR. However, doubling or halving the concentration of a disaccharide, which was used as a lyoprotectant, caused significant deviation between the NIR and Karl Fischer data because the NIR absorbance of the disaccharide overlapped with the moisture signal. Furthermore, complete removal of the disaccharide resulted in alteration of the protein NIR spectra, suggesting that NIR may be used to evaluate solid-state protein structure. The disaccharide concentration must be kept constant in this formulation to obtain accurate moisture results by NIR.


Nitric Oxide Inhalation Therapy Attenuates Postoperative Hypoxemia in Obese Patients with Acute Type A Aortic Dissection.
To investigate the differences between inhaled nitric oxide (iNO) treatment and conventional therapy in the treatment of postoperative hypoxemia in obese patients with acute type A aortic dissection (ATAAD). ATAAD patients diagnosed and treated with emergency surgery in our hospital from June 2017 to December 2019 were retrospectively analyzed. Patients with postoperative hypoxemia were divided into the iNO group and control group. Propensity score matching was used to analyze clinical characteristics and results of the two groups. A total of 218 ATAAD patients with BMI ≥ 25 were treated with surgery. Among them, 115 patients developed refractory hypoxemia (64 in the control group and 51 in the iNO group). Patients in the iNO group had significantly shorter invasive mechanical ventilation time, intensive care unit (ICU) stay, and hospital stay. After 6 h of iNO treatment, the PaO2/FiO2 ratio in the iNO group increased significantly, and this ratio was higher than that in the control group at 6, 12, 24, 48, and 72 h after treatment. Low-dose iNO could improve oxygenation and shorten mechanical ventilation and ICU stay in patients with hypoxemia after ATAAD surgery, but without significant side effects or increase in postoperative mortality or morbidity. These findings provide a basis for a randomized multicenter controlled trial to assess the efficacy of iNO in the treatment of hypoxemia after ATAAD surgery.


Ultrastructural study of development of hepatic necrosis induced by TNF-alpha and D-galactosamine.
Recent studies have suggested an association between tumor necrosis factor-alpha (TNF-alpha) and the development and progression of acute liver failure. To investigate the role of TNF-alpha in the mechanism of massive hepatic necrosis, we studied a mouse model of TNF-alpha and D-galactosamine (GalN) -induced hepatic necrosis by ultrastructural analysis. Administration of GalN caused edema of hepatocellular microvilli and widening of sinusoidal endothelial fenestrae (SEF); administration of TNF-alpha caused only a widening of the SEF. Massive hepatic necrosis with hemorrhage was seen 6 hr after concomitant administration of TNF-alpha and GalN. In the ultrastructural analysis, edema of the hepatocellular microvilli, widening of the SEF, and transmigration of red blood cells (RBC) and platelets to the space of Disse without exfoliation and necrosis of the sinusoidal endothelial cells were observed. Fibrin deposits were seen in areas adjacent to injured hepatocytes. The diameter of the SEF was significantly greater than in the nontreated group and the groups treated with TNF-alpha or GalN alone. These results suggest that as a consequence of the increase in diameter of the SEF, transmigration of RBCs and platelets to the space of Disse may have resulted in massive hepatic necrosis due to occlusion of the microcirculation.


An alternate pathway of arsenate resistance in E. coli mediated by the glutathione S-transferase GstB.
Microbial arsenate resistance is known to be conferred by specialized oxidoreductase enzymes termed arsenate reductases. We carried out a genetic selection on media supplemented with sodium arsenate for multicopy genes that can confer growth to E. coli mutant cells lacking the gene for arsenate reductase (E. coli ΔarsC). We found that overexpression of glutathione S-transferase B (GstB) complemented the ΔarsC allele and conferred growth on media containing up to 5 mM sodium arsenate. Interestingly, unlike wild type E. coli arsenate reductase, arsenate resistance via GstB was not dependent on reducing equivalents provided by glutaredoxins or a catalytic cysteine residue. Instead, two arginine residues, which presumably coordinate the arsenate substrate within the electrophilic binding site of GstB, were found to be critical for transferase activity. We provide biochemical evidence that GstB acts to directly reduce arsenate to arsenite with reduced glutathione (GSH) as the electron donor. Our results reveal a pathway for the detoxification of arsenate in bacteria that hinges on a previously undescribed function of a bacterial glutathione S-transferase.


[Clinical study of invasive fungal infection secondary to systemic lupus erythematosus].
To study the clinical characteristics of invasive fungal infection secondary to systemic lupus erythematosus (SLE). We observed the clinical features and experimental examination in 91 patients treated in Xiangya Hospital in recent years, of which 48 patients with invasive fungal infection and 41 patients without invasive fungal infection. The invasive fungal infection secondary to SLE mainly occurred in the lungs, nervous system, and urinary system. The fungi were mainly Candida albins and Aspergillus. The rate of invasive fungal infection in SLE patients and the level of CRP and TNF-α in these patients were significantly increased. The occurrence of invasive fungal infection was positively correlated with the prolonged course of disease, long-term use of immunosuppressants and antibiotics, and occurrence of complications, such as hypoproteinemia, leukocytopenia, and so on. The levels of C-reactive protein (CRP) and tumor necrosis factor-α(TNF-α) were increased in SLE patients with invasive fungal infection. The clinical features of SLE patients with invasive fungal infections are long course of disease, long-time use of immunosuppressants or antibiotics, and occurrence of complications, such as hypoproteinemia or leukopenia. The level of CRP and TNF-α can be used as an important reference index for diagnosing invasive fungal infections.


Transient rRNA synthesis inhibition with CX-5461 is sufficient to elicit growth arrest and cell death in acute lymphoblastic leukemia cells.
Enhanced rRNA synthesis is a downstream effect of many of the signaling pathways that are aberrantly activated in cancer, such as the PI3K/mTOR and MAP kinase pathways. Recently, two new rRNA synthesis inhibitors have demonstrated therapeutic effects on cancer cells while sparing normal cells. One of them, CX-5461, is currently in phase 1 clinical trials for hematological malignancies. Here, we investigate the effectiveness of transient treatment with this drug on acute lymphoblastic leukemia cells. Our results show that short exposure to CX-5461 followed by drug washout is sufficient to induce persistent G2 cell-cycle arrest and irreversible commitment to cell death, in spite of rRNA synthesis returning to normal within 24 hours of drug washout. The magnitude of cell death after transient exposure is similar to continuous exposure, but the time to cell death is relatively delayed with transient exposure. In this report, we also investigate rational drug combinations that can potentiate the effect of continuous CX-5461 treatment. We show that the checkpoint abrogator UCN-01 can relieve CX-5461-induced G2 arrest and potentiate the cytotoxic effects of CX-5461. Finally, we show that ERK1/2 is activated upon CX-5461 treatment, and that pharmacological inhibition of MEK1/2 leads to enhanced cell death in combination with CX-5461. In summary, our results provide evidence for the effectiveness of CX-5461 pulse treatment, which may minimize drug related toxicity, and evidence for enhanced effectiveness of CX-5461 in combination with other targeted agents.


Appraisal of testicular volumes: volumes matching ultrasound values referenced to stages of genital development.
Testicular volumes obtained with orchidometers or external linear measurements in the scrotum (centimeter ruler or calipers) grossly over-estimate ultrasound volumes, have much variability and may not be accurate or reproducible. The reference of the values obtained by orchidometers or US, to age or Tanner stages is not useful to determine the normal values for stages of puberty, because overlapping of ages and values. Pubertal development is determined by two events, genital and pubic hair development, that should be analyzed independently because one could be out of step with the other. The ultrasound (US) measurement of testicular volumes is the gold standard but is somewhat inconvenient, because it requires another procedure and, mainly, is costly. The solution of the problems would be to determine testicular volumes matching US values, from the width of the testis obtained in the scrotum with a centimeter ruler, by formulas recently described, and to reference them to the stages of genital development. The width and length of the testes in the scrotum with a centimeter ruler were obtained in 159 study subjects, in different stages of genital development and adults, for a total of 318 testicular determinations, from the age of 3 to 34 years. The width obtained in the scrotum was corrected by subtracting the values of the double scrotal skin (ss). The formulas were then applied and the testicular volumes matching US values were calculated. The volumes and the range of ages for different stages of genital development were determined. Penile measurements were obtained in 145 subjects and pubic and other hair recorded. Paired and unpaired 2 tail student t-test was used to compare the means of the different groups expressed as means and SD and, in addition the Wilcoxon rank sum test and Bootstrap methods for the testicular volume groups. A p value of 0.05 or less was considered significant. The Institutional Review Board (IRB) of Nationwide Children's Hospital determined that this study did not require IRB approval. With a simple measurement of the width of the testis in the scrotum, with a centimeter ruler, testicular volumes matching US values were calculated and normative values for each stage of genital development were determined. This information should solve present problems.


Effects of zafirlukast on capsular contracture: controlled study measuring the mammary compliance.
Capsular contracture is a highly distressing, difficult complication after breast augmentation for both the patient and the surgeon. Although capsular contracture is a multifactorial process, one common denominator in the successful treatment of this complication is believed to be the abatement of inflammation. Leukotriene antagonists have recently emerged as effective prophylactic agents in reactive airway diseases. Anecdotal reports have indicated that zafirlukast (Accolate, AstraZeneca) effectively reverses capsular contracture. A prospective study of capsular contracture in 120 female patients in whom a total of 216 prostheses were implanted was performed. The hardness of capsular contracture was assessed by means of the mammary compliance method (Anton Paar Mammacompliance system). The patients were divided into two groups: patients in group A received zafirlukast for a 6-month period, while those in group B received vitamin E. The results show a significant decrease of the values of breast compliance after 6 months in group A but not in group B and that the variation in compliance after 6 months in group A compared to group B is statistically significant. In zafirlukast-treated patients, we observed a reduction in mammary compliance of 7.69 percent after 1 month, 16.78 percent after 3 months and 24.01 percent after 6 months. The present study suggests that zafirlukast may be effective in reducing pain and breast capsule distortion in patients with longstanding contracture who are either not surgical candidates or who do not wish to undergo surgery.


Phase I study of third-line palliative chemotherapy with low dose paclitaxel for pancreatic cancer.
The prognosis of patients with unresectable or recurrent pancreatic cancers is very poor. Prior to development of nab-paclitaxel (PTX) plus gemcitabine (GEM) therapy and FOLFIRINOX therapy, there was no recommended third-line chemotherapy after 5-fluorouracil (5-FU) and GEM-based regimens. The present study conducted a Phase I clinical trial of weekly low-dose PTX as a third-line palliative chemotherapy for patients with pancreatic cancer. PTX was administered on days 1, 8, 15, and 22 of each cycle, repeated twice as follows: Level 1, 40 mg/m2 (n=6); Level 2, 50 mg/m2 (n=4). During the two cycles, three patients developed Grade 3 neutropenia in level 2; thus, the recommended dose was defined as 40 mg/m2. The disease control rate was 40.0% (stable disease, n=4). Median time to treatment failure of the four patients with stable disease was 5.5 months. In conclusion, palliative chemotherapy with low-dose PTX after failure of GEM and 5-FU is well tolerated and safe for unresectable or recurrent pancreatic cancer patients. The unique ID issues by UMIN: 000008148.


Hypothyroidism presenting as puzzling myalgias and cramps in 3 patients.
Cramps and myalgias have many etiologies. Three cases of hypothyroidism presenting with musculoskeletal symptoms are described. Each case was different in presentation, but all had no weakness but high creatine kinase levels. Treatment with thyroxine resolved all symptoms. These patients emphasize the importance of considering hypothyroidism in anyone with unexplained muscle symptoms.