HOXA3 accelerates wound healing in diabetic and aged non-diabetic mammals.
Chronic wounds are characterized by a persistent, hyper-inflammatory environment that prevents progression to regenerative wound closure. Such chronic wounds are especially common in diabetic patients, often requiring distal limb amputation, but occur in non-diabetic, elderly patients as well. Induced expression of HoxA3, a member of the Homeobox family of body patterning and master regulatory transcription factors, has been shown to accelerate wound closure in diabetic mice when applied topically as a plasmid encased in a hydrogel. We now provide independent replication of those foundational in vivo diabetic wound closure studies, observing 16% faster healing (3.3 mm wounds vs 3.9 mm wounds at Day 9 post original injury of 6 mm diameter) under treatment with observable microscopic benefits. We then expand upon these findings with minimal dose threshold estimation of 1 μg HoxA3 plasmid delivered topically at a weekly interval. Furthermore, we observed similarities in natural wound healing rates between aged non-diabetic mice and young diabetic mice, which provided motivation to test topical HoxA3 plasmid in aged non-diabetic mice. We observed that HoxA3 treatment achieved complete wound closure (0 mm diameter) at 2 weeks whereas untreated wounds were only 50% closed (3 mm wound diameter). We did not observe any gross adverse effects macroscopically or via histology in these short studies. Whether as a plasmid or future alternative modality, topical HoxA3 is an attractive translational candidate for chronic wounds.


Expression profiles of subtracted mRNAs during cellular senescence in human mesenchymal stem cells derived from bone marrow.
Cellular senescence is an irreversible cell cycle arrest that limits the replicative lifespan of cells. Senescence suppresses development of tumors by regulating aging factors, such as cyclin dependent kinase inhibitor (CKI) and telomerase. Suppression subtractive hybridization (SSH) was used to identify genes that were differentially expressed between young human mesenchymal stem cells (Y-hMSCs) and senescent human mesenchymal stem cells (S-hMSCs). We selected positive clones that were functionally characterized by referring to public databases using NCBI BLAST tool. This search revealed that 19 genes were downregulated, and 43 genes were upregulated in S-hMSCs relative to Y-hMSCs. Among subtracted clones in Y-hMSCs, most of genes markedly were related to metabolic functions. These genes, PDIA3, WDR1, FSTL1, COPG1, LMAN1, and PDIA6, significantly downregulated. Conversely, genes for subtracted clones in S-hMSCs were mostly associated with cell adhesion. In particular, the expression levels of 9 genes, HSP90B1, EID1, ATP2B4, DDAH1, PRNP, RAB1A, PGS5, TM4SF1 and SSR3, gradually increased during senescence. These genes have not previously been identified as being related to cellular senescence, but they seemed to be potentially affected during cellular senescence.


[Complex of single-stranded phage DNA and protein--a product of bacteriophage fl gene 3. Distribution of gene 3 protein in the DNA molecule].
Upon a 50% isopropanol treatment of phage fl in a 1 M NaCl solution protein A (gene 3 product)--DNA complex is precipitated while protein B (gene 8 product) was still solubilized. After such a treatment the DNA--protein complex containing 10--40% of protein A and less than 0.0025% of protein B was obtained. Evidence was obtained that there was no non-specific rearrangement of protein A during the isolation procedure. The complex was treated with endonuclease R.HAC III, followed by electrophoresis of the resulted fragments and estimation of the [14C] protein A (labeled with [14C]histidine) throughout the gel. The maximal radioactivity coincided with the DNA bands, being proportional to the DNA content in the respective bands. The data obtained indicate that protein A is iniformly arranged along the DNA molecule.


Treatment of Ventilator-Associated Pneumonia Using Intravenous Colistin Alone or in Combination with Inhaled Colistin in Critically Ill Children.
The objective of this study was to compare the safety and efficacy of inhaled plus intravenous (IV) colistin with that of IV colistin alone in critically ill children with ventilator-associated pneumonia (VAP) due to colistin-only susceptible (COS) Gram-negative bacteria (GNB). This retrospective cohort study included critically ill children aged 1 month to 18 years with culture-documented monomicrobial VAP due to COS GNB. Fifty patients were included, and 32 patients received IV colistin alone, whereas 18 patients received inhaled plus IV colistin. No between-cohort differences were observed in clinical (p = 0.49) and microbiological outcomes (p = 0.68), or VAP-related mortality (p = 0.99). Although the bacterial eradication rates did not differ in either treatment group, the median time to bacterial eradication (TBE) was significantly shorter in the inhaled plus IV colistin group than in the IV colistin group. The additional use of inhaled colistin was the only independent factor associated with TBE, and it shortened the median TBE by 3 days. Only one patient in the IV colistin group developed reversible nephrotoxicity. Mild bronchoconstriction was observed in three patients at the time of administration of the first doses of inhaled colistin, which did not require discontinuation of treatment. The present study has demonstrated that the addition of inhaled colistin to IV colistin led to a shorter TBE in critically ill children with VAP due to COS GNB. However, it did not lead to a significant difference in the clinical and microbiological outcomes of VAP.


The effects of iron supplementation on serum copper and zinc levels in pregnant women with high-normal hemoglobin.
To evaluate the effect of iron supplementation on serum copper and zinc levels. In a randomized, double-blind, placebo-controlled trial, 66 pregnant women with hemoglobin 13.2 g/dL or greater between the 13th and 18th week of pregnancy were randomized into case and control groups. From the 20th week until the end of pregnancy the case group received one ferrous sulfate tablet containing 50 mg elemental iron daily, while the control group received placebo. Hemoglobin, ferritin, copper, and zinc levels at 24-28 and 32-36 weeks of pregnancy were measured and compared. In the case group, serum copper levels in the second and third trimester were significantly lower than the control group (P<0.01 and P<0.001, respectively). Serum zinc levels in the case group in the second and third trimester were also significantly lower than the control group (P<0.001). Iron supplementation in pregnant women with hemoglobin greater than 13.2 g/dL reduces serum levels of copper and zinc.


An optimisation algorithm for determination of treatment margins around moving and deformable targets.
Determining treatment margins for inter-fractional motion of moving and deformable clinical target volumes (CTVs) remains a major challenge. This paper describes and applies an optimisation algorithm designed to derive such margins. The algorithm works by expanding the CTV, as determined from a pre-treatment or planning scan, to enclose the CTV positions observed during treatment. CTV positions during treatment may be obtained using, for example, repeat CT scanning and/or repeat electronic portal imaging (EPI). The algorithm can be applied to both individual patients and to a set of patients. The margins derived will minimise the excess volume outside the envelope that encloses all observed CTV positions (the CTV envelope). Initially, margins are set such that the envelope is more than adequately covered when the planning CTV is expanded. The algorithm uses an iterative method where the margins are sampled randomly and are then either increased or decreased randomly. The algorithm is tested on a set of 19 bladder cancer patients that underwent weekly repeat CT scanning and EPI throughout their treatment course. From repeated runs on individual patients, the algorithm produces margins within a range of +/-2 mm that lie among the best results found with an exhaustive search approach, and that agree within 3mm with margins determined by a manual approach on the same data. The algorithm could be used to determine margins to cover any specified geometrical uncertainty, and allows for the determination of reduced margins by relaxing the coverage criteria, for example disregarding extreme CTV positions, or an arbitrarily selected volume fraction of the CTV envelope, and/or patients with extreme geometrical uncertainties. An optimisation approach to margin determination is found to give reproducible results within the accuracy required. The major advantage with this algorithm is that it is completely empirical, and it is therefore particularly useful for CTVs where the geometrical uncertainties are difficult to model, such as the bladder.


Serial (1-3)-beta-D-Glucan (BDG) monitoring shows high variability among premature neonates.
This study aimed to characterize the baseline values and dynamics of serum (1,3)-Beta-D-Glucan (BDG) in neonates at high risk of neonatal invasive candidiasis (NIC); as well as to determine the effect of various clinical variables on these levels. Single center prospective cohort study was performed including 20 high-risk neonates (gestational age < 29 weeks and/or birth weight ≤ 1000 gr). Samples for BDG (Fungitell® assay) were obtained twice weekly during 6 weeks. Nineteen neonates were enrolled with a median gestational age of 25 weeks (IQR 24-27), median birth weight of 730 gr (IQR 650-810). None of the neonates was diagnosed with NIC. 190 serum samples were included. The median BDG value was 59 pg/ml (IQR 30-148), mean was 119 pg/ml (SD ± 154). A total of 42.1% (80/190) samples showed values ≥80 pg/ml, with all the neonates presenting at least one test above this cut-off. Neonatal age did not show an association with BDG levels. Exposure to steroids and the use of a heel prick as sampling method were associated with statistically significant higher BDG levels. The BDG levels showed high variability and in a significant proportion of samples values were above the threshold for positivity (e.g., ≥80 pg/ml) in the absence of NIC. The exposure to postnatal steroids and the heel prick as the method of blood sampling were associated with higher BDG levels. Neonatal invasive candidiasis (NIC) presents high morbi-mortality. The diagnosis of NIC is often challenging. Blood cultures have limitations and better diagnostic tools are needed. Beta-D-glucan is a diagnostic marker which could be potentially used, although still more clinical data are required.


[Respiratory muscle aids during an episode of aspiration in a patient with Duchenne muscular dystrophy].
We report the case of a Duchenne muscular dystrophy patient with good bulbar function but severely decreased forced vital capacity (9%) and spontaneous peak cough flow (PCF) (2.35 L/s). The patient needed continuous noninvasive ventilation (NIV) consisting of a volumetric ventilator with a nighttime nasal mask and a daytime mouthpiece. He also required application of manually assisted coughing techniques by insufflation with a resuscitation bag and chest thrust (manually assisted PCF after maximum insufflation capacity of 4.33 L/s). An episode of serious food aspiration was resolved by his main caregiver through NIV and manually assisted coughing. Bronchoscopy under sedation using NIV with a lip seal connection to his volumetric ventilator later revealed that no material remained. This case exemplifies the potential role of skilled respiratory management in some neuromuscular diseases.


Studies on Alternaria allergens. IV. Biologic activity of a purified Alternaria fraction (Alt-I).
We have previously isolated a purified Alternaria fraction (Alt-I) by means of immunochemical (RAST) assays to document allergenicity. In the present study we investigated the allergenicity of Alt-I by in vivo and in vitro biologic tests in a selected group of 16 nonimmunized Alternaria-sensitive patients, all of whom had positive skin tests to crude Alternaria extract; a control group of eight nonallergic individuals was also studied. Intradermal skin-test titration endpoints to Alt-I ranged from 60 pg/ml to 60 ng/ml in the Alternaria-sensitive patients. Alt-I induced greater than 30% in vitro leukocyte histamine in all Alternaria-sensitive patients at submicrogram concentrations. Alt-I did not produce positive skin tests or induce significant leukocyte histamine release in nonallergic individuals. In 14 of 16 Alternaria-sensitive patients bronchoprovocation testing produced greater than 20% fall in forced expiratory volume in one second at Alt-I test concentrations of 0.6 to 60 micrograms/ml. In a separate prospective study of 100 unselected individuals, concordant skin-test reactions were noted to crude Alternaria extract and Alt-I in 96 instances. These results confirm that the Alt-I fraction is a biologically active, major allergenic component of crude Alternaria extract.


Selection and constraint on regulatory elements in Drosophila simulans.
We utilized available Drosophila simulans molecular population genomic data to characterize sequence polymorphism in noncoding regulatory regions and their corresponding transcribed target genes or interacting transcription factors. We highlight two properties of regulatory evolution. First, we find that sequence divergence between D. melanogaster and D. simulans is greater at regulatory sites than expected from levels of intraspecific polymorphism, suggestive of positive selection. Second, we compared variation in cis-regulatory regions to that of the transcription factors and transcribed target genes with which they are associated. We find a correlation in polymorphism between cis-regulatory regions and the transcription factors which they bind, but not between cis-regulatory regions and the physically neighboring target genes which they directly regulate. This result indicates that the evolution of cis-regulatory elements is constrained by the interactions among transcription factors and regulatory regions. More broadly, these findings support the general importance of noncoding regulatory DNA in evolution.


Evaluation of top, angle, and side cleaned FIB samples for TEM analysis.
TEM specimens of a LaAlO(3)/SrTiO(3) multilayer are prepared by FIB with internal lift out. Using a Ga(+1) beam of 5 kV, a final cleaning step yielding top, top-angle, side, and bottom-angle cleaning is performed. Different cleaning procedures, which can be easily implemented in a dual beam FIB system, are described and compared; all cleaning types produce thin lamellae, useful for HRTEM and HAADF-STEM work up to atomic resolution. However, the top cleaned lamellae are strongly affected by the curtain effect. Top-angle cleaned specimens show an amorphous layer of around 5 nm at the specimen surfaces, due to damage and redeposition. Furthermore, it is observed that the LaAlO(3) layers are preferentially destroyed and transformed into amorphous material, during the thinning process.


Acrodermatitis enteropathica.
A 13-year-old girl presented with a history of red scaly plaques involving the chest, arms and legs beginning in infancy. Punch biopsy revealed psoriasiform hyperplasia and pallor of the epidermis. The patient's serum zinc level was 36 mug/dl [nl. 66-144 mug/dl]. A diagnosis of acrodermatitis enteropathica was established and the patient responded well to zinc replacement therapy. Acrodermatitis enteropathica is a rare autosomal recessive disorder caused by mutations in SLC39A4, which encodes the tissue-specific zinc transporter ZIP4.


ERK signalling and oncogene transformation are not impaired in cells lacking A-Raf.
Previous studies have indicated an important role for the Raf family of protein kinases in controlling cellular responses to extracellular stimuli and activated oncogenes, through their ability to activate the MEK/ERKs. To investigate the specific role of A-Raf in this process we generated A-Raf deficient mouse embryonic fibroblasts (MEFs) and embryonic stem (ES) cells by gene targeting and characterized their ability to undergo proliferation, differentiation, apoptosis, ERK activation, and transformation by oncogenic Ras and Src. The A-Raf deficient cells are not disrupted for any of these processes, despite the fact that this protein is normally expressed at high levels in both cell types. This implies either that A-Raf plays no role in MEK/ERK activation, that its function is fully compensated by other Raf proteins or MEK kinases or that its role in MEK/ERK activation is highly tissue-specific. Interestingly, B-Raf and Raf-1 activity towards MEK as measured by the immunoprecipitation kinase cascade assay are both significantly increased in the A-Raf deficient MEFs.


The children's brain tumor network (CBTN) - Accelerating research in pediatric central nervous system tumors through collaboration and open science.
Pediatric brain tumors are the leading cause of cancer-related death in children in the United States and contribute a disproportionate number of potential years of life lost compared to adult cancers. Moreover, survivors frequently suffer long-term side effects, including secondary cancers. The Children's Brain Tumor Network (CBTN) is a multi-institutional international clinical research consortium created to advance therapeutic development through the collection and rapid distribution of biospecimens and data via open-science research platforms for real-time access and use by the global research community. The CBTN's 32 member institutions utilize a shared regulatory governance architecture at the Children's Hospital of Philadelphia to accelerate and maximize the use of biospecimens and data. As of August 2022, CBTN has enrolled over 4700 subjects, over 1500 parents, and collected over 65,000 biospecimen aliquots for research. Additionally, over 80 preclinical models have been developed from collected tumors. Multi-omic data for over 1000 tumors and germline material are currently available with data generation for > 5000 samples underway. To our knowledge, CBTN provides the largest open-access pediatric brain tumor multi-omic dataset annotated with longitudinal clinical and outcome data, imaging, associated biospecimens, child-parent genomic pedigrees, and in vivo and in vitro preclinical models. Empowered by NIH-supported platforms such as the Kids First Data Resource and the Childhood Cancer Data Initiative, the CBTN continues to expand the resources needed for scientists to accelerate translational impact for improved outcomes and quality of life for children with brain and spinal cord tumors.


Applying genotoxicology tools to identify environmental stressors in support of river management.
Although bioassay approaches are useful for identifying chemicals of potential concern, they provide little understanding of the mechanisms of chemical toxicity. Without this understanding, it is difficult to address some of the key challenges that currently face aquatic ecotoxicology. To overcome this, the toxicity potential of the water samples was assessed and surviving organisms (Physa acuta) were used for protein activity measurements and gene expression profiling by making use of complementary DNA amplified fragment length polymorphism (cDNA-AFLP) analysis. From the data it was evident that the impacts of specific pollutants (e.g. sewage) on organisms at the cellular level could be identified, and that the expressed stressor genes can be used as bioindicators/markers/genetic signatures or fingerprints during identification of point source pollution. From an ecosystem management point of view these insights could assist with the forecasting and reduction of environmental risks on catchment level by implementing suitable management interventions.


Yeast ribosomal protein S3 possesses a β-lyase activity on damaged DNA.
Yeast ribosomal protein S3 has multifunctional activities that are involved in both protein translation and DNA repair. Here, we report that yeast Rps3p cleaves variously damaged DNA that contains not only AP sites and pyrimidine dimers but also 7,8-hydro-8-oxoguanine. This study also revealed that Rps3p has a β-lyase activity with a broad range of substrate specificity which cleaves phosphodiester bonds of UV or oxidatively damaged DNA substrates. Mutation analysis of the yeast Rps3 protein including introduction of domain deletions and residue replacements identified the residues Asp154 and Lys200 are important for the catalytic activity. In addition, the repair enzyme activity of yeast Rps3p was confirmed by complementation in xth, nfo Escherichia coli cells in which the DNA repair process is defective.


Biodegradation of nonylphenol in a continuous packed-bed bioreactor.
A packed bed bioreactor, with 170 ml glass bead carriers and 130 ml medium, was tested for the removal of the endocrine disrupter, nonylphenol, with a Sphingomonas sp. The bioreactor was first continuously fed with medium saturated with nonylphenol in an attempt to simulate groundwater pollution. At best, nonylphenol was degraded by 99.5% at a feeding rate of 69 ml h(-1) and a removal rate of 4.3 mg nonylphenol day(-1), resulting in a 7.5-fold decrease in effluent toxicity according to the Microtox. The bioreactor was then fed with soil leachates at 69 ml h(-1) from artificially contaminated soil (1 g nonylphenol kg(-1) soil) and a real contaminated soil (0.19 g nonylphenol kg(-1) soil). Nonylphenol was always completely removed from the leachates of the two soils. It was removed by 99% from the artificial soil but only 62% from real contaminated soil after 18 and 20 d of treatment, respectively, showing limitation due to nonylphenol adsorption.


Modeling Parkinson's disease-related symptoms in alpha-synuclein overexpressing mice.
Intracellular deposition of alpha-synuclein (α-syn) as Lewy bodies and Lewy neurites is a central event in the pathogenesis of Parkinson's disease (PD) and other α-synucleinopathies. Transgenic mouse models overexpressing human α-syn, are useful research tools in preclinical studies of pathogenetic mechanisms. Such mice develop α-syn inclusions as well as neurodegeneration with a topographical distribution that varies depending on the choice of promoter and which form of α-syn that is overexpressed. Moreover, they display motor symptoms and cognitive disturbances that to some extent resemble the human conditions. One of the main motives for assessing behavior in these mouse models is to evaluate the potential of new treatment strategies, including their impact on motor and cognitive symptoms. However, due to a high within-group variability with respect to such features, the behavioral studies need to be applied with caution. In this review, we discuss how to make appropriate choices in the experimental design and which tests that are most suitable for the evaluation of PD-related symptoms in such studies. We have evaluated published results on two selected transgenic mouse models overexpressing wild type (L61) and mutated (A30P) α-syn in the context of their validity and utility for different types of behavioral studies. By applying appropriate behavioral tests, α-syn transgenic mouse models provide an appropriate experimental platform for studies of symptoms related to PD and other α-synucleinopathies.


[Doppler ultrasound assisted hemorrhoid artery ligation. A new therapy in symptomatic hemorrhoids].
In 1995, Morinaga et al. (Japan) reported on a new technique in the treatment of hemorrhoids. We report the results of our first 105 patients thus treated. By a specially designed proctoscope coupled with a Doppler transducer, the hemorrhoidal arteries are looked for and ligated. All stages of hemorrhoid were treated. This method is painless, successful, and has a low rate of complications. It is for outpatients and is an alternative to all other methods in the treatment of hemorrhoids.


Digital advantage in the COVID-19 response: perspective from Canada's largest integrated digitalized healthcare system.
The SARS-CoV-2 pandemic has challenged healthcare systems worldwide. Uncertainty of transmission, limitations of physical healthcare system infrastructure and supplies as well as workforce shortages require dynamic adaption of resource deployment to manage rapidly evolving care demands, ideally based on real time data for the entire population. Moreover, shut down of traditional face-to-face care infrastructure requires rapid deployment of virtual health care options to avoid collapse of health organizations. The Alberta Electronic Health Record Information System is one of the largest population based comprehensive electronic medical record (EMR) installations. Alberta's long standing solid telehealth hardware-, training-, provider remuneration- and legislation infrastructure has enabled quick transition to virtual healthcare. Virtual health services including asynchronous secure clinical communications, real-time virtual care via messaging, telephony or video conferencing (telehealth) and ancillary functions like triage, scheduling, documentation and reporting, the previously established virtual hospital program with home monitoring, virtual health assessments, medication review, education and support for patients and families and coordination between family doctors, specialists and other health team members help to control viral transmission, protect healthcare personnel and save supplies. Moreover, rapid launch of online screening and triage tools to guide testing and isolation, online result sharing, infected patient and contact tracing including a smartphone exposure tracking application (ABTraceTogether), electronic best practice alerts and decision support tools, test and treatment order sets for standardized COVID-19 management, continuous access to population level real-time data to inform healthcare provider, public health and government decisions have become key factors in the management of a global crisis in Alberta.


Immunomodulatory effects of tretinoin in combination with clindamycin.
Since acne is a multifactorial skin disease, therapies affecting several etiologic factors can have a higher than expected effectiveness. A combination of the antibiotic clindamycin phosphate and the retinoic acid tretinoin was developed. Anti-inflammatory and immunomodulatory effects of tretinoin in vitro were studied on human keratinocytes and peripheral blood mononuclear cells (PBMCs). Effects of clindamycin phosphate on tretinoin effects were studied. Anti-inflammatory effects on keratinocytes were assessed using an in vitro model with PMA (phorbol ester)-stimulated A431 cells (human epidermoid carcinoma). Immunomodulatory effects were measured on superantigen (SEB) stimulated PBMCs. Tretinoin showed very potent inhibition of PMA-stimulated IL-6 (interleukin 6) release by A431 cells. The addition of clindamycin phosphate did not interfere with this effect. Tretinoin very potently stimulated IL-5 release, and inhibited IFN gamma release by SEB-stimulated human PBMCs. This indicates an immunomodulatory effect, stimulating Th2, and inhibiting Th1 dominated responses. These features have been related to the healing of acne lesions. The addition of clindamycin phosphate did not interfere with the immunomodulatory effects of tretinoin. The combination of tretinoin and clindamycin phosphate can be expected to be very effective in acne therapy.


Functional significance of central D1 receptors in cognition: beyond working memory.
The role of dopamine D1 receptors in prefrontal cortex function, including working memory, is well acknowledged. However, relatively little is known about their role in other cognitive or emotional functions. We measured both D1 and D2 receptors in the brain using positron emission tomography in healthy subjects, with the aim of elucidating how regional D1 and D2 receptors are differentially involved in cognitive and emotional functions beyond working memory. We found an inverted U-shaped relation between prefrontal D1 receptor availability and Wisconsin Card Sorting Test performance, indicating that too little or too much D1 receptor stimulation impairs working memory or set shifting. In addition, variability of D1 receptor availability in the amygdala and striatum was related to individual differences in emotional responses and decision-making processes, respectively. These observations suggest that the variability of available D1 receptors might be associated with individual differences in brain functions that require phasic dopamine release. An interdisciplinary approach combining molecular imaging of dopamine neurotransmission with cognitive neuroscience and clinical psychiatry will provide new perspectives for understanding the neurobiology of neuropsychiatric disorders such as schizophrenia, addiction and Parkinson's disease, as well as novel therapeutics for cognitive impairments observed in them.


iTRAQ-based quantitative proteomic analysis reveals potential factors associated with the enhancement of phenazine-1-carboxamide production in Pseudomonas chlororaphis P3.
Phenazine-1-carboxamide (PCN), a phenazine derivative, is strongly antagonistic to fungal phytopathogens. Pseudomonas chlororaphis HT66 is a PCN-producing, non-pathogenic biocontrol strain, and we obtained the mutant P. chlororaphis P3, which produces 4.7 times more PCN than the wild-type HT66 strain. To reveal the cause of PCN production enhancement in P3 and find potential factors related to PCN biosynthesis, an iTRAQ-based quantitative proteomic analysis was used to study the expression changes between the two strains. Of the 452 differentially expressed proteins, most were functionally mapped into PCN biosynthesis pathway or other related metabolisms. The upregulation of proteins, including PhzA/B, PhzD, PhzF, PhzG, and PhzH, involved in PCN biosynthesis was in agreement with the efficient production of PCN in P3. A number of proteins that function primarily in energy production, amino acid metabolism, and secondary metabolism played important roles in PCN biosynthesis. Notably, proteins involved in the uptake and conversion of phosphate, inorganic nitrogen sources, and iron improved the PCN production. Furthermore, the type VI secretion system may participate in the secretion or/and indirect biosynthetic regulation of PCN in P. chlororaphis. This study provides valuable clues to better understand the biosynthesis, excretion and regulation of PCN in Pseudomonas and also provides potential gene targets for further engineering high-yield strains.


Human umbilical cord hyaluronate. Neutral sugar content and carbohydrate-protein linkage studies.
Paper chromatography of neutral sugars and gas chromatography of their aldononitrile acetates indicated the presence of fucose, arabinose and a small amount of glucose in purified human umbilical cord hyaluronate. The molar ratios of serine, threonine and aspartic acid to neutral sugars were not unity, suggesting the non-involvement of the neutral sugars and the amino acids in a carbohydrate-protein linkage. The same was indicated by an increase in the percentage of the aforementioned amino acids and by the absence of sugar alditols in umbilical cord hyaluronate reduced eith NaBH4 -PdCl2, after alkali treatment. This reduction caused a decrease in the intrinsic viscosity and molecular wieght to about one-half and an appreciable decrease in the specific rota tion of hyaluronate, suggesting a separation of the two antiparallel chains o the double helical hyaluronate. The umbilical cord hyluronate containe contained bound silicon and it is possible that this bound silicon may cross-link the two chains at interspersed intervals through the uronic acid moiety and/or through neutral sugars.


Nano-photoluminescence of natural anyon molecules and topological quantum computation.
The proposal of fault-tolerant quantum computations, which promise to dramatically improve the operation of quantum computers and to accelerate the development of the compact hardware for them, is based on topological quantum field theories, which rely on the existence in Nature of physical systems described by a Lagrangian containing a non-Abelian (NA) topological term. These are solid-state systems having two-dimensional electrons, which are coupled to magnetic-flux-quanta vortexes, forming complex particles, known as anyons. Topological quantum computing (TQC) operations thus represent a physical realization of the mathematical operations involving NA representations of a braid group Bn, generated by a set of n localized anyons, which can be braided and fused using a "tweezer" and controlled by a detector. For most of the potential TQC material systems known so far, which are 2D-electron-gas semiconductor structure at high magnetic field and a variety of hybrid superconductor/topological-material heterostructures, the realization of anyon localization versus tweezing and detecting meets serious obstacles, chief among which are the necessity of using current control, i.e., mobile particles, of the TQC operations and high density electron puddles (containing thousands of electrons) to generate a single vortex. Here we demonstrate a novel system, in which these obstacles can be overcome, and in which vortexes are generated by a single electron. This is a ~ 150 nm size many electron InP/GaInP2 self-organized quantum dot, in which molecules, consisting of a few localized anyons, are naturally formed and exist at zero external magnetic field. We used high-spatial-resolution scanning magneto-photoluminescence spectroscopy measurements of a set of the dots having five and six electrons, together with many-body quantum mechanical calculations to demonstrate spontaneous formation of the anyon magneto-electron particles (eν) having fractional charge ν = n/k, where n = 1-4 and k = 3-15 are the number of electrons and vortexes, respectively, arranged in molecular structures having a built-in (internal) magnetic field of 6-12 T. Using direct imaging of the molecular configurations we observed fusion and braiding of eν-anyons under photo-excitation and revealed the possibility of using charge sensing for their control. Our investigations show that InP/GaInP2 anyon-molecule QDs, which have intrinsic transformations of localized eν-anyons compatible with TQC operations and capable of being probed by charge sensing, are very promising for the realization of TQC.


Mechanical Performance of Polymeric ARGF-Based Fly Ash-Concrete Composites: A Study for Eco-Friendly Circular Economy Application.
At present, low tensile mechanical properties and a high carbon footprint are considered the chief drawbacks of plain cement concrete (PCC). At the same time, the combination of supplementary cementitious material (SCM) and reinforcement of fiber filaments is an innovative and eco-friendly approach to overcome the tensile and environmental drawbacks of plain cement concrete (PCC). The combined and individual effect of fly ash (FA) and Alkali resistance glass fiber (ARGF) with several contents on the mechanical characteristics of M20 grade plain cement concrete was investigated in this study. A total of 20 concrete mix proportions were prepared with numerous contents of FA (i.e., 0, 10, 20, 30 and 40%) and ARGF (i.e., 0, 0.5, 1 and 1.5%). The curing of these concrete specimens was carried out for 7 and 28 days. For the analysis of concrete mechanical characteristics, the following flexural, split tensile, and compressive strength tests were applied to these casted specimens. The outcomes reveal that the mechanical properties increase with the addition of fibers and decrease at 30 and 40% replacement of cement with fly ash. Replacement of cement at higher percentages (i.e., 30 and 40) negatively affects the mechanical properties of concrete. On the other hand, the addition of fibers positively enhanced the flexural and tensile strength of concrete mixes with and without FA in contrast to compressive strength. In the end, it was concluded that the combined addition of these two materials enhances the strength and toughness of plain cement concrete, supportive of the application of an eco-friendly circular economy. The relationship among the mechanical properties of fiber-reinforced concrete was successfully generated at each percentage of fly ash. The R-square for general relationships varied from (0.48-0.90) to (0.68-0.96) for each percentage of FA fiber reinforced concrete. Additionally, the accumulation of fibers effectively boosts the mechanical properties of all concrete mixes.


Direct evidence of caeruloplasmin antioxidant properties.
The chain-breaking antioxidant potential of caeruloplasmin and bovine serum albumin (BSA) has been investigated in comparison with other well-established antioxidants. Their Oxygen Radical Absorbing Capacity (ORAC), was measured by using beta-phycocyanin (beta-PC) as a fluorescent indicator protein, 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH) as a peroxyl radical generator and the water soluble vitamin E analogue, Trolox, as a reference standard. The relative peroxyl absorbing capacities/mole for Trolox, caeruloplasmin, heat-denatured caeruloplasmin (hCP), catalase, bovine serum albumin (BSA), superoxide dismutase (SOD), and deferoxamine were 1; 2.6; 3.3; 3.7; 1.2; 0.1; 0.2, respectively. Caeruloplasmin was far more effective as a peroxyl radical scavenger than SOD, deferoxamine and BSA, but slightly less effective than catalase. The peroxyl radical absorbing capacity of caeruloplasmin was enhanced by heat-denaturation of the protein. Electron paramagnetic resonance (EPR) spectroscopy using 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as a spin-trap, was applied in order to measure the scavenger abilities of caeruloplasmin on superoxide radical and hydroxyl radical production and the concentration required to inhibit by 50% oxygen free radical formation (IC50) was determined. The IC50 values of caeruloplasmin, hCP, and BSA for the superoxide radical were 12, 2, 260 microM and for the hydroxyl radical 15, 2, 200 microM. These results show that caeruloplasmin is an effective chain-breaking antioxidant for a variety of radicals, independently of its catalytic ferroxidase activity.


Fine mapping of the restorer gene Rfp3 from an Iranian primitive rye (Secale cereale L.).
A comparative genetics approach allowed to precisely determine the map position of the restorer gene Rfp3 in rye and revealed that Rfp3 and the restorer gene Rfm1 in barley reside at different positions in a syntenic 4RL/6HS segment. Cytoplasmic male sterility (CMS) is a reliable and striking genetic mechanism for hybrid seed production. Breeding of CMS-based hybrids in cereals requires the use of effective restorer genes as an indispensable pre-requisite. We report on the fine mapping of a restorer gene for the Pampa cytoplasm in winter rye that has been tapped from the Iranian primitive rye population Altevogt 14160. For this purpose, we have mapped 41 gene-derived markers to a 38.8 cM segment in the distal part of the long arm of chromosome 4R, which carries the restorer gene. Male fertility restoration was comprehensively analyzed in progenies of crosses between a male-sterile tester genotype and 21 recombinant as well as six non-recombinant BC4S2 lines. This approach allowed us to validate the position of this restorer gene, which we have designated Rfp3, on chromosome 4RL. Rfp3 was mapped within a 2.5 cM interval and cosegregated with the EST-derived marker c28385. The gene-derived conserved ortholog set (COS) markers enabled us to investigate the orthology of restorer genes originating from different genetic resources of rye as well as barley. The observed localization of Rfp3 and Rfm1 in a syntenic 4RL/6HS segment asks for further efforts towards cloning of both restorer genes as an option to study the mechanisms of male sterility and fertility restoration in cereals.


Using the Resident Assessment Instrument-Mental Health (RAI-MH) to determine levels of care for individuals with serious mental illness.
This paper outlines the development of an algorithm to determine appropriate levels of care (LOC) for individuals with a serious mental illness (SMI). The algorithm, drew on several domains of the Resident Assessment Instrument-Mental Health (RAI-MH) to support a statistical model that would explain a maximum of variance with the gold standard, a consensus-based global rating of required LOC. The RAI-MH model explained 67.5% of the variance. The validity of the model was further examined by determining how the discrepancy between the current and predicted levels of care related to psychiatric outcomes. The results demonstrated that undersupported clients experienced significant negative psychiatric outcomes compared to clients receiving adequate care. Although the model based on the RAI-MH is not perfect, the results warrant further research to determine its usefulness in predicting required LOC.


Contribution of a locking cross-stitch to a modified kessler core suture: a new technique.
We compared the gap-producing forces, ultimate tensile strength, and slip resistance of a new core suture technique and that of a modified Kessler suture in freshly harvested chicken tendons. Biomechanical trials were carried out using a tensile-strength testing machine. Our technique proved superior to the modified Kessler suture with respect to its ultimate tensile strength and slip resistance.


Have driving restrictions reduced air pollution: evidence from prefecture-level cities of China.
With rapid economic development, air pollution became a serious problem in China. Vehicle's exhaust emissions are among the main sources of air pollution. With samples of 173 prefecture-level cities in China from 2006 to 2016, this paper takes the implementation of driving restrictions as a quasi-natural experiment to examine the relationship between driving restrictions and air quality through the difference-in-differences method based on the propensity score matching. Results indicate that driving restrictions have not improved the air quality in terms of average treatment effect as there is no clear evidence that people turn from private cars to busses or subways. Furthermore, heterogeneous effects of driving restrictions exist across different regions and cities of China. The implementation of driving restrictions has significantly positive impacts on air pollution in the Eastern and Central cities of China, while it has significantly negative effects in non-capital cities. This study implies that the development of local public transportation needs to be taken into account when formulating the policy of driving restrictions.


Repaglinide-loaded nanostructured lipid carriers with different particle sizes for improving oral absorption: preparation, characterization, pharmacokinetics, and in situ intestinal perfusion.
Repaglinide-loaded nanostructured lipid carriers (REP-NLCs) with different particle sizes were successfully designed and prepared to investigate the permeation and absorption ability by in situ single-pass intestinal perfusion (SPIP) study and pharmacokinetics. Both of the formulations prepared by solvent diffusion method exhibited a spherical shape under transmission electron microscopy (TEM) and similar zeta potential value of -11 mV. The particles size, encapsulation efficiency (EE), drug loading (DL) of REP-NLCs-Small and REP-NLCs-Large size preparations were about 79 nm and 325 nm, 96.83% and 98.60%, 4.41% and 3.05%, respectively. Besides, both REP-NLCs showed good colloidal stability and had no burst release phenomenon compared with REP-Sol. SPIP demonstrated the improved membrane permeability for NLCs compared with REP-Sol, especially NLCs-Small size preparation. The bioavailability increased sequentially in REP-Sol, REP-NLCs-Large, and REP-NLCs-Small, and the difference between each other was statistical significant. Our investigations demonstrate that NLCs with small particles size of 50-100 nm, such as 79 nm, are able to enhance absorption performance of a poorly soluble repaglinide compared with large particles size, such as 325 nm, by significantly improving the absorption in jejunum, and colon of rats and thus well improving oral bioavailability.


Genomic profiling of malignant phyllodes tumors reveals aberrations in FGFR1 and PI-3 kinase/RAS signaling pathways and provides insights into intratumoral heterogeneity.
Malignant phyllodes tumors of the breast are poorly understood rare neoplasms with potential for aggressive behavior. Few efficacious treatment options exist for progressed or metastatic disease. The molecular features of malignant phyllodes tumors are poorly defined, and a deeper understanding of the genetics of these tumors may shed light on pathogenesis and progression and potentially identify novel treatment approaches. We sequenced 510 cancer-related genes in 10 malignant phyllodes tumors, including 5 tumors with liposarcomatous differentiation and 1 with myxoid chondrosarcoma-like differentiation. Intratumoral heterogeneity was assessed by sequencing two separate areas in 7 tumors, including non-heterologous and heterologous components of tumors with heterologous differentiation. Activating hotspot mutations in FGFR1 were identified in 2 tumors. Additional recurrently mutated genes included TERT promoter (6/10), TP53 (4/10), PIK3CA (3/10), MED12 (3/10), SETD2 (2/10) and KMT2D (2/10). Together, genomic aberrations in FGFR/EGFR PI-3 kinase and RAS pathways were identified in 8 (80%) tumors and included mutually exclusive and potentially actionable activating FGFR1, PIK3CA and BRAF V600E mutations, inactivating TSC2 mutation, EGFR amplification and PTEN loss. Seven (70%) malignant phyllodes tumors harbored TERT aberrations (six promoter mutations, one amplification). For comparison, TERT promoter mutations were identified by Sanger sequencing in 33% borderline (n=12) and no (0%, n=8) benign phyllodes tumors (P=0.391 and P=0.013 vs malignant tumors, respectively). Genetic features specific to liposarcoma, including CDK4/MDM2 amplification, were not identified. Copy number analysis revealed intratumoral heterogeneity and evidence for divergent tumor evolution in malignant phyllodes tumors with and without heterologous differentiation. Tumors with liposarcomatous differentiation revealed more chromosomal aberrations in non-heterologous components compared with liposarcomatous components. EGFR amplification was heterogeneous and present only in the non-heterologous component of one tumor with liposarcomatous differentiation. The results identify novel pathways involved in the pathogenesis of malignant phyllodes tumors, which significantly increase our understanding of tumor biology and have potential clinical impact.


Climate Change, Pollution, Deforestation, and Mental Health: Research Trends, Gaps, and Ethical Considerations.
Climate change, pollution, and deforestation have a negative impact on global mental health. There is an environmental justice dimension to this challenge as wealthy people and high-income countries are major contributors to climate change and pollution, while poor people and low-income countries are heavily affected by the consequences. Using state-of-the art data mining, we analyzed and visualized the global research landscape on mental health, climate change, pollution and deforestation over a 15-year period. Metadata of papers were exported from PubMed®, and both relevance and relatedness of terms in different time frames were computed using VOSviewer. Co-occurrence graphs were used to visualize results. The development of exemplary terms over time was plotted separately. The number of research papers on mental health and environmental challenges is growing in a linear fashion. Major topics are climate change, chemical pollution, including psychiatric medication in wastewater, and neurobiological effects. Research on specific psychiatric syndromes and diseases, particularly on their ethical and social aspects is less prominent. There is a growing body of research literature on links between mental health, climate change, pollution, and deforestation. This research provides a graphic overview to mental healthcare professionals and political stakeholders. Social and ethical aspects of the climate change-mental health link have been neglected, and more research is needed.


A source of cutaneous maternal semiochemicals in the mink?
Unique hypertrophic apocrine sweat glands are described in the neck, perineal and inguinal skin of mink kits. These glands enlarge after birth, only to regress rapidly and become vestigal by weaning. No similar phenomenon has been recognized before in mammals. Behavioral studies indicate a possible role for the glandular secretion in maternal recognition of the young.


Use of accumulation images obtained by arterial-phase contrast-enhanced harmonic grayscale ultrasonography to evaluate tumor vessels in focal nodular hyperplasia and other hepatic tumors.
To evaluate the usefulness of accumulation images obtained by arterial-phase contrast-enhanced harmonic grayscale ultrasonography for examining tumor vessels in focal nodular hyperplasia and other hepatic lesions. After injecting a galactose-palmitic acid contrast agent, we used conventional contrast-enhanced harmonic grayscale ultrasonography to scan 8 focal nodular hyperplasia lesions, 21 hepatocellular carcinomas, 2 cholangiocellular carcinomas, 12 hepatic metastases, 17 hemangiomas, and 2 angiomyolipomas. We then accumulated and superimposed consecutive conventional images (accumulation images) and compared them with corresponding conventional images to evaluate serial images of hepatic tumor vessels. Serial tumor vessel findings obtained from accumulation images were superior to those obtained from conventional images in 51 (77%) of the 66 various hepatic lesions examined. Evidence of the spoke-wheel artery pattern in 2 of the 8 focal nodular hyperplasia lesions was equivocal in the conventional images, but accumulation images clearly depicted serial images of spoke-wheel arteries, clearly indicating a diagnosis of focal nodular hyperplasia. Accumulation images allowed the diagnosis of two additional focal nodular hyperplasia lesions that had not been correctly diagnosed as focal nodular hyperplasia from conventional images. Accumulation images obtained by arterial-phase contrast-enhanced harmonic grayscale ultrasonography are useful in evaluating hepatic tumor vessels, especially spoke-wheel arteries, which specifically indicate focal nodular hyperplasia.


Distinct α-Synuclein:Lipid Co-Structure Complexes Affect Amyloid Nucleation through Fibril Mimetic Behavior.
A hallmark of Parkinson's disease is the presence of Lewy bodies consisting of lipids and proteins, mainly fibrillated α-synuclein (aSN). aSN is an intrinsically disordered protein exerting its physiological role in an ensemble of states, one of which coexists in large assemblies with lipids, recently termed co-structures. Here, we decipher the kinetics of aSN:lipid co-structure formation to decode its mechanism of formation, and we show that the co-structures form with a distinct stoichiometry. Through seeded fibrillation assays, we demonstrate that aSN:lipid co-structures accelerate aSN fibril nucleation compared to lipid vesicles alone. A small-angle X-ray scattering-based model is proposed in which aSN decorates the lipid vesicle surface, yielding properties similar to those of the fibril surface, enhancing fibril nucleation. The delicate balance of aSN structural states close to and on the membrane may under given conditions, e.g., increased local concentrations, be a crucial switching factor between functional and pathological behavior.


Helicobacter pylori infection is associated with reduced circulating ghrelin levels independent of body mass index.
Ghrelin stimulates growth hormone and has orexigenic and adipogenic effects. Plasma ghrelin levels are reduced in obesity and possibly in Helicobacter pylori infection. To investigate whether there was a relation between H. pylori infection, body mass index (BMI) and serum ghrelin or leptin levels. University students undergoing an annual health check-up were invited to participate. H. pylori status was based on the presence of specific IgG H. pylori antibodies in urine. Fasting serum ghrelin, leptin levels, and pepsinogen I and II levels were measured by enzyme immunoassay (EIA). Eight hundred and one students volunteered. There was no significant difference in the height and BMI between those with and without H. pylori infection. The population of ghrelin study consisted of 132 (66 H. pylori-positive and 66 H. pylori-negative) students matched for age, sex, and BMI. The ghrelin level in the H. pylori-positive group was significantly lower (median 55 pmol/l) compared to the H. pylori-negative group (103 pmol/l) (p < .00001). Leptin, triglyceride, total cholesterol, and HDL-cholesterol were not different between the two groups, whereas LDL-cholesterol levels were significantly higher (106 versus 100 mg/dl) (p = .03) in the H. pylori-positive group. Leptin levels correlated with the BMI (r = 0.53) (p < .00001). Among H. pylori-positive subjects, ghrelin correlated only with pepsinogen I levels (r = 0.26, p = .04). H. pylori infection was associated with a reduction in circulating ghrelin levels independent of sex and BMI.


The role of substance P in the axon reflex in the rat.
The increase in vascular permeability in small blood vessels of rat skin induced by antidromic stimulation of sensory roots, nerves, or single identified polymodal nociceptive fibres or by intra-arterial injection of Substance P, has been examined by light and electron microscopy. The effects of local application of capsaicin (a Substance P-depleting agent) to both skin and muscle have also been studied. Nerve stimulation and Substance P induce leakage of a venular pattern similar to that caused by known permeability factors. However, capsaicin causes leakage from both venules and capillaries in a pattern more characteristic of direct endothelial injury. It is suggested that neurogenic leakage may be mediated by local liberation of Substance P, but caution is necessary in interpreting experiments involving the use of capsaicin because of its local toxic effects.


Full-term live birth in a woman with 17α-hydroxylase and 17,20-lyase deficiency with assisted reproductive technology: a case report.
17α-hydroxylase deficiency, which is caused by a CYP17A1 gene mutation, is a rare type of congenital adrenocortical hyperplasia that mainly manifests as hypertension, hypokalaemia and sexual dysplasia. To date, few pregnancies associated with this syndrome have been reported. We describe a 35-year-old Chinese woman with nonclassical congenital adrenal hyperplasia (NCCAH) due to 17α-hydroxylase/17,20-lyase deficiency who achieved pregnancy after in vitro fertilization (IVF) and frozen-thawed embryo transfer. She had secondary amenorrhea since she was 27, and subsequently, high level of progesterone in the follicular phase was found during a blood test. A compound heterozygous mutation was found in the CYP17A1 gene, c.1263G > A and c.985_987delinsAA. The patient was given standardized treatment with dexamethasone. Due to ovulation disorder, IVF was performed. She underwent whole embryo vitrification freezing. Frozen-thawed embryo transplantation was performed following the artificial cycle protocol of endometrium preparation, resulting in a singleton pregnancy. At 39 weeks and 1 day of gestation, caesarean section was performed due to the breech position of the foetus. A high level of progesterone reduces endometrial receptivity. Standardized treatment with dexamethasone and frozen-thawed embryo transfer with an artificial cycle protocol of endometrium preparation should be the choice for infertile female patients with CYP17A1 deficiency.


Pterygium excision with free conjunctival autograft (FCG) versus postoperative strontium 90 (90Sr) beta-irradiation. A prospective study.
In a prospective study the results of a pterygium excision in 54 patients (57 eyes) who underwent a superficial free conjunctival autograft (FCG) were compared to those of patients who were treated with postoperative 90Sr-irradiation. In 51 cases the minimum follow-up was six months, the maximum follow-up seven years. We divided the study up into a randomized part and an open part. In the randomized part, surgery of a primary pterygium was performed in 25 eyes, of which 16 were treated with a FCG and compared with 9 eyes with primary pterygium surgery and postoperative 90Sr beta-irradiation. In the same period 16 eyes were treated because of a recurrent pterygium: 8 with FCG and 8 with 90Sr-irradiation. In the open part of the study 16 eyes with primary pterygium were successively treated with FCG alone. The results showed in the randomized, as well as in the open study on primary surgery with a minimum follow-up of six month, one recurrence in each of the FCG-groups (2 out of 31 eyes = 6.4%), and no recurrences in the 90Sr-group (0%). In the randomized group of patients treated for a recurrent pterygium one recurrence developed in the FCG group (1 out of 8 eyes = 12.5%) and one in the 90Sr-group (1 out of 7 = 14.6%). Analysis of other clinical parameters showed that postoperative treatment with corticosteroids, nonsteroidal anti-inflammatory drugs, and artificial tears was necessary for a longer period in the FCG group than in the 90Sr-group.


Gastroduodenal intussusception due to Peutz-Jeghers syndrome in infancy.
A case of Peutz-Jeghers Syndrome (PJS) presenting in infancy with gastric-outlet obstruction is described. PJS may become symptomatic at any age and should be suspected when there are obstructive symptoms or gastrointestinal blood loss in a baby with a positive family history. Contrast studies and endoscopy are useful in diagnosis and surveillance. Treatment requires a combination of endoscopy and laparotomy/laparoscopy. Because of the reported cancer risk, we recommend surveillance from the age of diagnosis.


Expression, purification and osteogenic bioactivity of recombinant human BMP-4, -9, -10, -11 and -14.
Bone morphogenetic proteins (BMPs) are cytokines from the TGF-beta superfamily, with important roles during embryonic development and in the induction of bone and cartilage tissue differentiation in the adult body. In this contribution, we report the expression of recombinant human BMP-4, BMP-9, BMP-10, BMP-11 (or growth differentiation factor-11, GDF-11) and BMP-14 (GDF-5), using Escherichia coli pET-25b vector. BMPs were overexpressed, purified by affinity his-tag chromatography and shown to induce the expression of early markers of bone differentiation (e.g. smad-1, smad-5, runx2/cbfa1, dlx5, osterix, osteopontin, bone sialoprotein and alkaline phosphatase) in C2C12 cells and in human adipose stem cells. The described approach is a promising method for producing large amounts of different recombinant BMPs that show potential for novel biomedical applications.


Experimental verification of the half-life of 65Zn.
The half-life of 65Zn was determined experimentally and compared with literature values. Two ampoules of a 65Zn solution were measured at regular intervals in two ionisation chambers over a period corresponding to nearly two half-lives. The result of 243.8+/-0.3 days is in agreement with the value of 244.0+/-0.2 days obtained by De Roost et al. (Z. Phys. 250 (1975) 395) at the same institute (previously called CBNM) in 1972. Moreover, this half-life is supported by most of the other measurements reported in the literature.


Distribution ratios of ethanol and water between whole blood, plasma, serum, and erythrocytes: Recommendations for interpreting clinical laboratory results in a legal context.
This article reviews the scientific literature dealing with the distribution of ethanol and water between whole blood (WB), plasma, serum, and erythrocytes (red-blood cells). Knowledge of the ethanol distribution ratio is important when analytical results derived from hospital clinical laboratories are interpreted in a forensic context, such as during the prosecution of traffic offenders. Statutory blood-alcohol concentration (BAC) limits for driving are defined as the concentration of ethanol in WB and not in plasma, serum or red-blood cells. These bio-fluids differ in their water content and thereby the concentrations of ethanol. Plasma and serum contain ~90%-92% w/w water, WB ~78%-80% w/w and erythrocytes ~64%-66% w/w. The mean plasma/WB and serum/WB distribution ratios of ethanol are therefore expected to be ~1.15:1 (91/79 = 1.15), which is in good agreement with values determined empirically. However, in individual cases, the actual distribution ratio will depend on the person's age, gender, and biochemical and hematological properties of the blood specimen, such as its hematocrit. For legal purposes, we recommend that the concentration of ethanol in plasma or serum determined at hospital laboratories is divided by a factor of 1.2, which would provide a conservative estimate of the co-existing BAC and the chance of overestimating the true value is only 1 in 2000 (0.05%).


Early detection of Biscogniauxia nummularia in symptomless European beech (Fagus sylvatica L.) by TaqMan quantitative real-time PCR.
To develop a quantitative real-time PCR (Rt PCR) assay for the early detection of Biscogniauxia nummularia, a xylariaceous fungus that causes strip-canker and wood decay on European beech (Fagus sylvatica L.). The molecular assay was based on TaqMan chemistry using species-specific primers and a fluorogenic probe designed on the ITS1 sequence of rRNA gene clusters. The specificity of the oligonucleotides and the probe were tested using the DNA of B. nummularia isolates from different geographic areas, of phylogenetically related species, and of some fungi commonly colonizing European beech bark and wood. A total of 31 symptomless and symptomatic shoots of European beech were collected from three forest sites in the Apennine Mountains of Italy. The percentage of positive detections of B. nummularia with the TaqMan assay was 78.6%, compared with only 14.3% of positive isolations on growth media for two sites. In shoots, the quantitative Rt PCR assay detected down to 8.0-fg fungal DNA per microgram of total DNA extracted. The assay developed in quantitative Rt PCR, by using TaqMan chemistry, revealed a rapid and sensitive method useful for the early detection of B. nummularia in symptomless European beech twigs.


A case report of a giant rectal adenoma causing secretory diarrhea and acute renal failure: McKittrick-Wheelock syndrome.
The McKittrick-Wheelock syndrome is a rare depletion syndrome caused by a secretory villous adenoma or a carcinoma of the rectosigmoid tract. An aggressive hydroelectrolyte rebalancing is often needed, and curative treatment is obtained only with complete removal of the lesion, by endoscopy or surgery. Low clinical suspicion often delays the diagnosis, resulting in detrimental complications. We report the case of a 75-year-old woman, presenting to the emergency department with acute renal failure and electrolyte imbalance, reporting an history of recurrent episodes of dehydration and chronic diarrhea. After being admitted to the nephrology department she underwent diagnostic investigation that revealed the presence of a giant adenoma of the rectum. The patients received supportive therapy and was subsequently treated with surgery, with a favorable outcome. A prompt diagnosis plays an important role in the treatment of McKittrick-Wheelock syndrome. We describe a case of this condition in detail and review the related literature, underlining the typical diagnostic features and exploring the possible therapeutic options.


Five commercially-available antibodies react differentially with allelic forms of human HLA-DR beta chain.
Allelic variants of HLA-DRB1 have been associated with a variety of autoimmune and infectious diseases. Although the precise molecular mechanisms by which HLA-DRB1 alleles predispose to a particular disease are currently unclear, it has been shown that mRNA expression levels of HLA-DRB1 are dependent on the different alleles. We aimed to measure HLA-DR beta chain levels in peripheral blood mononuclear cells of individuals carrying HLA-DRB1*03:01/*04:01 and HLA-DRB1*03:01/*15:01 alleles by western blotting, using five commercially-available HLA-DRB antibodies. We observed highly heterogeneous binding of the tested antibodies to the different allelic forms of the HLA-DR beta chain. Overall, we show that current immunological research that employs available antibodies to detect HLA-DR beta chains is biased towards detection of specific variants of the protein; this may cause significant discrepancy in quantification of protein expression in a heterogeneous human population.


Is surgical duration associated with postoperative complications in primary shoulder arthroplasty?
Surgical duration is an independent predictor of short-term adverse outcomes after a variety of orthopedic procedures, both arthroscopic and open. However, this association in shoulder arthroplasty remains unclear. The purpose of this study was to identify the association between surgical duration and postoperative complications, as well as increased use of health care resources, after shoulder arthroplasty. Primary shoulder arthroplasty procedures performed from 2005 to 2016 were identified in the American College of Surgeons National Surgical Quality Improvement Program database using Current Procedural Terminology codes. Surgical duration was divided into 3 cohorts: (1) surgical procedures lasting less than 90 minutes, (2) those lasting between 90 and 120 minutes, and (3) those lasting more than 120 minutes. Baseline patient characteristics and outcome variables were compared using bivariate analysis. Outcome variables were compared using multivariate analysis. Overall, 14,106 patients were identified. Longer surgical duration was significantly associated with younger age, male patients, higher body mass index, and use of general anesthesia, (P < .001 for each), as well as smoking history (P < .39). Relative to operative times shorter than 90 minutes, surgical procedures lasting more than 120 minutes had higher rates of any complication (P = .002), return to the operating room (P = .008), urinary tract infection (P = .02), non-home discharge (P < .001), blood transfusion (P < .001), and unplanned 30-day hospital readmission (P = .03). Increasing surgical duration was associated with a variety of postoperative medical complications and increased use of health care resources including discharge to acute care facilities, blood transfusions, and hospital readmission. These data suggest that surgical duration should be considered for postoperative risk stratification, as well as patient counseling, and may be a surgeon-modifiable risk factor independent of patient risk factors.


Oral etodolac, a COX-2 inhibitor, reduces postoperative pain immediately after fast-track cardiac surgery.
The present study was designed to evaluate the efficacy of a cyclooxygenase (COX)-2 inhibitor, etodolac, on postoperative pain after fast-track cardiac surgery, and to examine the changes in plasma etodolac concentration after oral administration. Thirty patients scheduled for elective coronary artery bypass grafting (CABG) surgery were randomly assigned preoperatively in a double-blind fashion to receive either vehicle ( n = 15) or etodolac 400 mg ( n = 15) via a gastric tube at the end of the surgery. Standardized fast-track cardiac anesthesia was used. In both groups, postoperative pain was treated with buprenorphine suppository. Visual analogue pain scores (VASs) were recorded immediately after extubation and at 24 h after surgery. Plasma etodolac concentration was measured at 1, 2, and 6 h after administration ( n = 8). No difference was detected in time to extubation between the etodolac group (209 +/- 85 min, mean +/- SD) and the vehicle group (207 +/- 98 min). VASs were significantly lower in the etodolac (2.3 +/- 2.1) vs the vehicle group (5.8 +/- 2.0) immediately after extubation ( P = 0.009), but no difference was detected in pain scores at 24 h after surgery, or in the amount of buprenorphine administered in the intensive care unit (ICU), or in the incidence of side effects. Plasma etodolac concentration was within the pharmaceutically recommended range at 1 h, 2 h, and 6 h after administration. The oral use of etodolac with rectal buprenorphine reduces pain scores immediately after cardiac surgery without an increase in side effects.


Gain at chromosomal region 5p15.33, containing TERT, is the most frequent genetic event in early stages of non-small cell lung cancer.
Chromosomal imbalances resulting in altered gene dosage play a role in the molecular pathogenesis of non-small cell lung cancer (NSCLC), but the target genes remain to be identified. To identify early-stage genetic events that drive progression of NSCLC, we conducted a high-resolution array comparative genomic hybridization (CGH) study, using an array of 4,046 bacterial artificial chromosome clones to screen for DNA copy number changes associated with individual genes in 36 tumors obtained from patients in early stages of NSCLC. Multiple early genetic events occurring on chromosome 5p were identified, with a minimal detection region at 5p15.33 approximately 12. The most frequent finding involved gain of 5p15.33, observed in 15 of 19 stage I (A+B) cancers (79%) and in 28 of the total 36 NSCLC cases (78%). This locus harbors the genes TERT, SLC6A19, and SLC6A18 and is a telomeric boundary at bacterial artificial chromosome (BAC) clone 91_J20. Other potential candidate genes evidencing high numbers of genomic copy number changes (> or =40% of patients) included the following genes, encountered in >50% of 19 stage I (A+B) cancers: CEP72 and TPPP (14 of 19; 74%); AHRR, EXOC3 (previously SEC6L1), SLC9A3, LOC442126, ZDHHC11, BRD9, and TRIP13 (13/19; 68%); and CLPTM1L (alias CRR9), SLC6A3 (previously DAT1), and LOC401169 (10/19; 53%). Fluorescence in situ hybridization validated the array CGH findings. The gain of 5p15.33 is thus one of the most consistent alterations in the early stages of lung cancer, and a series of genes in the critical 5p15.33 region may be used as novel biomarkers for the early detection and classification of lung cancer.


Immunocytochemical localization of secretory component in Paneth cell secretory granules-rat Paneth cells participate in acquired immunity.
With the marker of Paneth cells-lysozyme, secretory component (SC) immunoreactivity was demonstrated exclusively in Paneth cells of rat small intestine. The other types of epithelial cells (columnar, goblet, endocrine) were negative. On electron microscopic level, many SC-positive colloidal gold particles were found in rough endoplasmic reticulum, Golgi complexes, basal membrane and secretory granules of Paneth cells. These results suggest that SC is not a component of ingested immune complex, but a membrane receptor on Paneth cell. It may function as receptor for polymeric IgA and mediate its transport across the mucosal epithelium. Thus, Paneth cells are responsible for SC synthesis and participate in IgA-mediated acquired immunity in rat small intestine.


Facilitation of adenoviral wild-type p53-induced apoptotic cell death by overexpression of p33(ING1) in T.Tn human esophageal carcinoma cells.
To investigate the effect of p33(ING1) on wild-type p53 gene therapy, T.Tn human esophageal carcinoma cells were stably transfected with p33(ING1) cDNA. Infection with Ad-p53 (recombinant adenovirus containing wild-type p53) into p33-transfected cells reduced cell viability, while infection with empty vector had little effect. This reduced viability was shown to be due to apoptotic cell death by the TUNEL (terminal deoxynucleotidyl transferase-mediated nick end-labeling) assay. Following infection with Ad-p53, levels of p53 were similar in p33-expressing cells and in the parental line. However, levels of p21 and Mdm2 were elevated in p33-transfected cells. Nonetheless, this enhanced expression of Mdm2 appeared to be ineffective in downregulating p53. Transient transfection with mutant Mdm2 prior to Ad-p53 infection provided a significant protection as compared with cells transfected with wild-type Mdm2. These results imply a synergistic effect between p33 and p53 in the induction of apoptosis of human esophageal carcinoma cells. A role for Mdm2 in this synergism is suggested.


Observation and quantification of aberrant crypts in the murine colon treated with a colon carcinogen: preliminary findings.
In the present study a methodological approach is taken which quantitates aberrant dysplastic crypts in the unsectioned murine colon. C57BL/6J or CF1 female mice (7-8 weeks old) were injected (i.p.) with azoxymethane (5 mg/kg body wt./week) for 4 weeks. Their colons were excised, cut open on the median axis and fixed flat in buffered formalin. Unsectioned colons were stained with methylene blue. The mucosal side was examined under a light microscope. The aberrant crypts, which are larger and have a thicker epithelial lining, were easily visualized using X 4 or X 10 objectives. CF1 mice, which are more sensitive to developing colon tumors, had a higher number of aberrant crypts/colon than their less sensitive counterparts, C57BL/6J mice (5.0 +/- 0.7 vs. 2.4 +/- 0.7). The usefulness of this observation as a possible measure of neoplastic events is discussed in the animal and human situation.


Role of delayed interval debulking for persistent residual disease after more than 5 cycles of chemotherapy for primary advanced ovarian cancer. An international multicenter study.
Standard of care in patients with advanced ovarian cancer (AOC) is upfront surgery followed by chemotherapy. Neoadjuvant chemotherapy (NACT) and interval debulking surgery (IDS) is an alternative in selected patients. Most data exist with IDS following 3-4 cycles chemotherapy, however, some patients experience a delay of IDS. So far, the impact of a "delayed" interval debulking surgery (DID) is poorly defined. We analyzed data from eight international gynecology-oncology referral centers. Patients were included if they had newly diagnosed AOC and were prone to DID (minimum 5 cycles of NACT) between 2011 and 2017. 308 patients underwent DID. 89.6% had a high-grade serous ovarian cancer. The median number of pre-op NACT was 6 cycles (range 5-9) and 6.1% of patients received additionally bevacizumab. The majority of patients had stage-IV disease (51.3%). Median duration of surgery was 210 min (range 34-561), the median surgical complexity score was 4 (range 1-16). Complete resection was achieved in 60.1%. The median number of post-op chemotherapy cycles was 2 (range 0-5). The rate of severe complications (Clavien-Dindo£3°) was 9.7% and 30 days post-op mortality was 0.3%. The median PFS and OS in patients with complete resection was 19.5 and 49.2 months compared to 14.8 and 33.0 months in patients with incomplete resection (p = 0.001), respectively. We did not observe any survival benefit for patients with cytoreduction to small residuals (1-10 mm) compared to residual disease >1 cm. Our data may suggest that offering surgery to patients with persistent disease after 5+ cycles could be associated with favorable outcome if a complete resection is achieved. Patients who had residual disease postoperatively may experience rather peri-operative treatment burden than any benefit from DID.


Toxicological effects of three different types of highly pure graphene oxide in the midge Chironomus riparius.
Graphene oxide (GO) is a carbon nanomaterial used in electronics, biomedicine, environmental remediation and biotechnology. The production of graphene will increase in the upcoming years. The carbon nanoparticles (NPs) are released into the environment and accumulated in aquatic ecosystems. Information on the effects of GO in aquatic environments and its impact on organisms is still lacking. The aim of this study was to synthesise and characterise label-free GO with controlled lateral dimensions and thickness - small GO (sGO), large GO (lGO) and monolayer GO (mlGO) - and determine their impact on Chironomus riparius, a sentinel species in the freshwater ecosystem. Superoxide dismutase (SOD) and lipid peroxidation (LPO) was evaluated after exposures for 24 h and 96 h to 50, 500, and 3000 μg/L. GOs accumulated in the gut of C. riparius and disturbed its antioxidant metabolism. We suggest that all types of GO exposure can upregulate of SOD. Moreover, both lGO and mlGO treatments caused LPO damage in C. riparius in comparison to sGO, proving its favourable lateral size impact in this organism. Our results indicate that GOs could accumulate and induce significant oxidative stress on C. riparius. This work shows new information about the potential oxidative stress of these NMs in aquatic organisms.


Chemical properties of technetium-99m-DL-homocysteine, a possible tumor-imaging agent.
The chemical properties of 99mTc-DL-homocysteine (99mTc-Hcy) showing high accumulation in several experimental tumors were investigated. The form of tumor-tropic 99mTc-Hcy was a polymeric complex which appeared at void volume on Sephadex G-15 by eluting with 5 mM Hcy. This complex changed into smaller complexes of ca. 600 molecular weight in the presence of 150 mM NaCl and 5 mM Hcy, suggesting that 99mTc-Hcy was a complex composed of smaller polymers which are weakly bound together by an ionic bond. The complex showed a negative charge. The Hcy/Tc molar ratio in the complex was approximately 2 and no Sn was detected.


Development of a paper-based lateral flow immunoassay for simultaneous detection of lipopolysaccharides of Salmonella serovars.
Lateral flow type detection is becoming interesting not only in regions with a poor medical infrastructure but also for practitioners in day-to-day clinical work or for veterinary control in case of possible epidemics. In this work, we describe the first steps of development of a multi-channel strip with potential internal calibration of multiparametric and colorimetric lateral flow assays for the simultaneous detection of the lipopolysaccharides (LPS) of Salmonella typhimurium (S. typhimurium) and Salmonella enteritidis (S. enteritidis). We structured four channels in the nitrocellulose membrane with a Yb:KGW solid-state femtosecond laser ("cold" ablation process) to form distinct tracks of porous material and used gold nanoparticles for the labeling of the antibodies. In addition, calibration curves of the spot intensities of both serovars are presented, and it was shown that no cross reactivity between the different capture antibodies and LPS occurred. Finally, we detected LPS of both Salmonella serovars simultaneously. The color changes (spot intensities of the reaction zones) were evaluated using the open-source image-processing program ImageJ. Graphical abstract Multiparametric testing, strip A was tested with LPS S. enteritidis ( c=0.01 g/L) and LPS S.typhimurium ( c=0.0001 g/L), strip B with LPS S. enteritidis ( c=0.001 g/L) and LPS S. typhimurium ( c=0.001g/L) and strip C with LPS S. enteritidis (c=0.0001 g/L) and LPS S. typhimurium ( c=0.01 g/L), and read-out.


Publisher Correction: Room-temperature superconductivity in a carbonaceous sulfur hydride.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Corticosteroid Dosing and Glucose Levels in COPD Patients Are Not Associated with Increased Readmissions.
Hospital admissions and readmissions for chronic obstructive pulmonary disease (COPD) exacerbations are associated with increased mortality and higher cost. The management of exacerbations with a shortened course of systemic corticosteroids has similar efficacy as compared to longer steroid courses, but actual overall steroid dose given is still variable. The outcomes associated with steroid side effects, such as hyperglycemia, need further evaluation. We hypothesized that the use of higher doses of corticosteroids, and the subsequent hyperglycemia, contributes to readmission. This is a retrospective study at a tertiary care referral center in central Texas between February 2014 and July 2016. Daily corticosteroid dose, blood glucose levels, and readmission rates at 30 and 31-90 days were recorded. Sample characteristics are described using descriptive statistics. A chi-square test or student's t-test were used to test for associations in bivariate comparisons. Multivariable logistic regression assessed the association between readmission rate and demographic and clinical characteristics. There were 1120 patients admitted for COPD exacerbation between February 2014 and July 2016. A total of 57% were female, mean age was 69 years (standard deviation [SD] 12), and average body mass index (BMI) was 29.4 (SD 9.8). Of the total, 349 (31%) had diabetes prior to admission. The 30-day readmission rate was 16%, and the readmission rate from 31-90 days was 14%. The average prednisone equivalent dose per day during hospitalization was 86 mg (SD 52). A multivariable logistic regression model did not show any significant association between readmission and average daily glucose, high maximum glucose (>180 mg/dL on any reading), or prednisone equivalent administered per day. Corticosteroid dose and hyperglycemia were not associated with an increased 30-day or 31-90-day readmission rate after COPD exacerbation discharge. In addition, using higher doses of corticosteroids instead of standard-of-care (prednisone 40 mg per day for a 5-day period) did not appear to affect the readmission rate in this cohort.


Neonatal aphakia is associated with altered levels of dopamine metabolites in the non-human primate retina.
Neonatal aphakia is associated with retardation of the axial elongation of the neonatal eye. In contrast, form deprivation increases axial elongation, an effect that has been associated with decreased retinal dopamine metabolism. The present investigation was conducted to test the hypothesis that neonatal aphakia induces an effect on the levels of retinal dopamine opposite to form deprivation. Lensectomy and vitrectomy were performed on the right eyes of rhesus monkeys at approximately 1 week of age; their left eyes were unmanipulated. Axial length was measured by A-scan ultrasonography. Prior to surgery, mean axial length of the right and left eyes was identical. Following lens removal, both eyes continued to elongate, however the aphakic eyes elongated at a slower rate resulting in a significant shorter axial length compared to that of the unmanipulated eye. Removal of the crystalline lens had no effect on steady-state dopamine levels in either central or peripheral retina. However, levels of the dopamine metabolites, 3,4-dihydroxyphenylacetic acid and homovanillic acid were significantly elevated in central retina, but not in the peripheral retina of aphakic eyes. Our results support the hypothesis that dopamine is a component of the retinal signaling pathways that are involved in the regulation of eye growth and emmetropization.


Enzymatic changes in peripheral blood leukocytes in rats in subacute benzene vapours poisoning. II. The activity of neutrophils enzymes.
The activity of some enzymes of neutrophils in peripheral blood of rats in subacute benzene vapour poisoning (an exposure to the concentration of benzene vapour of 27.000 mg/m3, 6 hr daily for 10 consecutive days) was determined cytochemically. Increase in the activity of alkaline phosphatase and decrease in that of acid hydrolases: acid phosphatase, beta-glucuronidase and beta-glucosaminidase were found. The results obtained indicate destructive action of benzene on lysosomes of peripheral blood neutrophils.


Evaluating the impact of pass/fail United States Medical Licensing Examination Step 1 scoring on pathology residency selection.
Beginning 26 January 2022, the United States Medical Licensing Examination Step 1 changed from a numerical score to Pass/Fail. Historically, residency programs have used Step 1 scores as a valuable metric in assessing the competitiveness of applicants. We assessed how residency program criteria will change when evaluating applicants after Step 1 becomes Pass/Fail. A survey was distributed to the program directors of all 144 pathology residency programs accredited by Accreditation Council for Graduate Medical Education. Survey questions evaluated the importance of using Step 1 and Step 2 Clinical Knowledge (CK) scores when assessing applicants. Participants were asked to rank a list of applicant criteria used before and after Step 1 becomes Pass/Fail. Data were analyzed using chi-squared and paired t-tests with significance at P < 0.05. A total of 34 residency program directors (23.6%) responded to the survey. 76.5% (P< 0.001) of responders believed Step 1 scores were able to predict a resident's ability to pass their board exams, while 41.2% believed Step 2 CK could predict a resident's ability to pass board exams and perform clinically in pathology (P = 0.282). 61.8% of responders agreed that an applicant's medical school ranking would become more important (P = 0.001). There were no significant differences in the relative importance of 16 selection criteria after the change of Step 1 to Pass/Fail. It does not appear that Step 2 CK will become more important. Although results are constrained by a 23.6% response rate, it can be a start to guiding future students through residency applications.


Glyoxal causes inflammatory injury in human vascular endothelial cells.
To explore mechanisms of diabetes-associated vascular endothelial cells (ECs) injury, human umbilical vein ECs were treated for 24h with high glucose (HG; 26mM), advanced glycation end-products (AGEs; 100mug/ml) or their intermediate, glyoxal (GO: 50-5000muM). HG and AGEs had no effects on ECs morphology and inflammatory states as measured by vascular cell adhesion molecule (VCAM)-1 and cyclooxygenase (COX)-2 expressions. GO (500muM, 24h) induced cytotoxic morphological changes and protein expression of COX-2 but not VCAM-1. GO (500muM, 24h) activated ERK but not JNK, p38 or NF-kappaB. However, ERK inhibitor PD98059 was ineffective to GO-induced COX-2. While EUK134, synthetic combined superoxide dismutase/catalase mimetic, had no effect on GO-mediated inflammation, sodium nitroprusside inhibited it. The present results indicate that glyoxal, a metabolite of glucose might be a more powerful inducer for vascular ECs inflammatory injury. Nitric oxide but not anti-oxidant is preventive against GO-mediated inflammatory injury.


Trefoil Factor Family 1 Inhibits the Development of Hepatocellular Carcinoma by Regulating β-Catenin Activation.
Recent studies have suggested that trefoil factor family 1 (TFF1) functions as a tumor suppressor in gastric and pancreatic carcinogenesis. To investigate the role of TFF1 in hepatocarcinogenesis, we performed immunohistochemical staining of surgically resected human liver samples, transfected a TFF1 expression vector into hepatocellular carcinoma (HCC) cell lines, and employed a mouse model of spontaneous HCC development (albumin-cyclization recombination/Lox-Stop-Lox sequence-Kirsten rat sarcoma viral oncogene homologG12D [KC]); the model mouse strain was bred with a TFF1-knockout mouse strain to generate a TFF1-deficient HCC mouse model (KC/TFF1-/- ). TFF1 expression was found in some human samples with HCC. Interestingly, TFF1-positive cancer cells showed a staining pattern contradictory to that of proliferating cell nuclear antigen, and aberrant DNA hypermethylation in TFF1 promoter lesions was detected in HCC samples, indicating the tumor-suppressive role of TFF1. In vitro, induction of TFF1 expression resulted in impaired proliferative activity and enhanced apoptosis in HCC cell lines (HuH7, HepG2, and HLE). These anticancer effects of TFF1 were accompanied by the loss of nuclear β-catenin expression, indicating inactivation of the β-catenin signaling pathway by TFF1. In vivo, TFF1 deficiency in KC mice accelerated the early development and growth of HCC, resulting in poor survival rates. In addition, immunohistochemistry revealed that the amount of nuclear-localized β-catenin was significantly higher in KC/TFF1-/- mice than in KC mice and that human HCC tissue showed contradictory expression patterns for β-catenin and TFF1, confirming the in vitro observations. TFF1 might function as a tumor suppressor that inhibits the development of HCC by regulating β-catenin activity.


Protocol for generation of 3D bone marrow surrogate microenvironments in a rotary cell culture system.
In this protocol, we describe how to generate 3D culture surrogates of chronic lymphocytic leukemia (CLL) and multiple myeloma (MM) bone marrow microenvironments. We detail the use of culturing scaffolds populated with BM stromal cells and tumor cells in the RCCS™ bioreactor. This 3D culture can efficiently recapitulate tumor-stroma crosstalk and allows the testing of drugs such as ibrutinib and bortezomib. Moreover, this protocol can be used for the generation of other and more complex tumor microenvironments. For complete details on the use and execution of this protocol, please refer to Belloni et al. (2018) and Barbaglio et al. (2021).


Floristic composition and vegetation analysis in Hail region north of central Saudi Arabia.
In this study, 19 sites representing different habitats in Hail region were regularly visited for two years, in each site 2-5 stands were selected for investigating floristic composition and vegetation types in the area. A total of 124 species representing 34 families were recorded. The family Asteraceae is represented by the highest number of species (21 species) followed by the Poaceae (17 species) and the Brassicaceae (10 species) whereas, 15 families including Acanthaceae, Convolvulaceae, Moraceae, Nyctaginaceae and Primulaceae, are represented by a single species each. Chronological analysis of the vegetation in the area revealed the domination of Saharo-Sindian elements in the wild vegetations and of weedy species in the cultivated plots. Therophytes and chamaephytes are the dominating life forms of the vegetation spectra; therophytes represent 49.20% and chamaephytes represent 29.00% of the total species in the study area. Application of TWINISPAN and DECORANA classification and ordination techniques to the data produced seven vegetation groups. Ruderal habitats comprised two small groups A and F dominated by Phragmites australis and Imperata cylindrical (A), Euphorbia peplus and Sisymbrium irio (F), respectively. Two vegetation groups (B and G) have been recognized in the mountains and slopes dominated by Launaea mucronata, Trigonella stellata (B) and Ficus palmate and Fagonia bruguieri (G). Other two groups (C and E) inhabit the desert and mountainous wadies; these are represented by Gymnocarpos decandrus and Ochradenus baccatus (C) and Senecio glaucus subsp. coronopifolius and Rumex equisetiforme (E). On the other hand, one group (D) inhabits the cultivated plots and is represented by Plantago albicans and Rumex vesicarius, the last group also includes species restricted to the sand dune habitat of the Al-Nafud desert north of Hail city and represented by Calligonum polygonoides and Halyxolon salicornicum. The vegetation analysis indicated the invasion of Hail Flora by some foreign weeds such as Solanum nigrum, Lactuca serriola and Amaranthus lividus. The presence of these weeds points out the need to monitor the vegetation change in Hail region, and also other regions of Saudi Arabia, in order to elucidate the human impact on the wild plants diversity as human activities change with the fast development in the kingdom.


Recombinant factor VIIa use in acute type A aortic dissection repair: A multicenter propensity-score-matched report from the Nordic Consortium for Acute Type A Aortic Dissection.
Surgery for acute type A aortic dissection (ATAAD) is often complicated by excessive bleeding. Recombinant factor VIIa (rFVIIa) effectively treats refractory bleeding associated with ATAAD surgery; however, adverse effects of rFVIIa in these patients have not been fully assessed. Here we evaluated rFVIIa treatment in ATAAD surgery using the Nordic Consortium for Acute Type A Aortic Dissection (NORCAAD) database. This was a multicenter, propensity score-matched, retrospective study. Information about rFVIIa use was available for 761 patients, of whom 171 were treated with rFVIIa. We successfully matched 120 patients treated with rFVIIa with 120 controls. Primary endpoints were in-hospital mortality, postoperative stroke, and renal replacement therapy (RRT). Survival data were presented using Kaplan-Meier estimates. Compared with controls, patients treated with rFVIIa received more transfusions of packed red blood cells (median, 9.0 U [4.0-17.0 U] vs 5.0 U [2.0-11.0 U]; P = .008), platelets (4.0 U [2.0-8.0 U] vs 2.0 U [1.0-4.4 U]; P <.001), and fresh frozen plasma (8.0 U [4.0-18.0 U] vs 5.5 U [2.0-10.3 U]; P = .01) underwent reexploration for bleeding more often (31.0% vs 16.8%; P = .014); and had greater 24-hour chest tube output (1500 L [835-2500 mL] vs 990 mL [520-1720 mL]). Treatment with rFVIIa was not associated with significantly increased rates of in-hospital mortality (odds ratio [OR], 0.74; 95% confidence interval [CI], 0.34-1.55; P = .487), postoperative stroke (OR, 1.75; 95% CI, 0.82-3.91; P = .163), or RRT (OR, 1.18; 95% CI, 0.48-2.92; P = .839). In this propensity-matched cohort study of patients undergoing ATAAD surgery, treatment with rFVIIa for major bleeding was not associated with a significantly increased risk of stroke, RRT, or mortality.


Thrombocytopenia and insufficient thrombopoietin production in human small-for-gestational-age infants.
Small-for-gestational-age (SGA) infants are at increased risk for transient thrombocytopenia. The aim of this study was to determine whether thrombocytopenia in human SGA infants is due to insufficient thrombopoietin (TPO) production. A prospective study of 202 infants with gestational age less than 37 weeks was conducted; 30 of them were SGA infants, and 172 were non-SGA infants. Thrombocytopenia was seen in 17 of 30 SGA infants and 40 of 172 non-SGA infants. Platelet counts were significantly lower in the SGA group than in the non-SGA group at the time of the lowest platelet count within 72 h of birth. The platelet count and immature platelet fraction (IPF) were negatively correlated in non-SGA infants, but not in SGA infants. In addition, the platelet count and TPO were negatively correlated in non-SGA infants. IPF and TPO were significantly lower in SGA than in non-SGA infants with thrombocytopenia. IPF increased with thrombocytopenia to promote platelet production in non-SGA infants due to increasing TPO, but not in SGA infants. This study found an association between insufficient TPO production and thrombocytopenia in SGA infants. In addition, this study is important for understanding the etiology of thrombocytopenia in SGA infants. The immature platelet fraction was low, and serum thrombopoietin was not increased in small-for-gestational-age (SGA) infants with thrombocytopenia. Thrombocytopenia in SGA infants is due to insufficient thrombopoietin production. This study is important for understanding the etiology of thrombocytopenia in SGA infants.


Risk stratification and hierarchy of antenatal care.
Cardiac disease is the leading cause of maternal death in the UK. The triennial maternal mortality reports have repeatedly highlighted failure to recognise the level of risk as a major contributing factor to the deaths of these women. Once the level of risk has been recognised, services then need to be organised in a way that supports the needs of the highest risk women, but avoids unnecessary intervention for women at lower risk. Risk scoring systems and lesion-specific indicators may help predict maternal and neonatal outcomes. Care can then be planned accordingly, to optimise the outcome for the woman and her baby.


Polycomb gene product Bmi-1 regulates stem cell self-renewal.
The Polycomb group (PcG) gene Bmi-1 has recently been implicated in the maintenance of hematopoietic stem cells (HSCs). However, the role of each component of PcG complex in HSCs and the impact of forced expression of PcG genes on stem cell self-renewal remain to be elucidated. To address these issues, we performed both loss-of-function and gain-of-function analysis on various PcG proteins. Expression analysis revealed that not only Bmi-1 but also other PcG genes are predominantly expressed in HSCs. Loss-of-function analyses, however, demonstrated that absence of Bmi-1 is preferentially linked with a profound defect in HSC self-renewal, indicating a central role for Bmi-1, but not the other components, in the maintenance of HSC self-renewal. Over-expression analysis of PcG genes also confirmed an important role of Bmi-1 in HSC self-renewal. Our findings indicate that the expression level of Bmi-1 is the critical determinant for the self-renewal capacity of HSCs. These findings uncover novel aspects of stem cell regulation exerted through epigenetic modifications by the PcG proteins.


Whole-Genome Sequence of "Candidatus Liberibacter asiaticus" from Guangdong, China.
The draft genome sequence of "Candidatus Liberibacter asiaticus" strain A4, isolated from a mandarin citrus in Guangdong, People's Republic of China, is reported. The A4 strain has a genome size of 1,208,625 bp, G+C content of 36.4%, 1,107 predicted open reading frames, and 53 RNA genes.