Investigation of the covalent binding of styrene-7,8-oxide to DNA in rat and mouse.
Styrene-7,8-oxide (SO), the main intermediate metabolite of styrene, induces hyperkeratosis and tumors in the forestomach of rats and mice upon chronic administration by gavage. The aim of this study was to investigate whether DNA binding could be responsible for the carcinogenic effect observed. [7-3H]SO was administered by oral gavage in corn oil to male CD rats at two dose levels (1.65 or 240 mg/kg). After 4 or 24 h, forestomach, glandular stomach and liver were excised, DNA was isolated and its radioactivity determined. At the 4 h time point, the DNA radioactivity was below the limit of detection in the forestomach and the liver. Expressed in the units of the covalent binding index, CBI = (mumol adduct/mol DNA nucleotide)/(mmol chemical administered/kg body wt), the DNA-binding potency was below 2.6 and 2.0 respectively. In the glandular stomach at 4 h, and in most 24 h samples, DNA was slightly radiolabeled. Enzymatic degradation of the DNA and separation by HPLC of the normal nucleotides showed that the DNA radioactivity represented biosynthetic incorporation of radiolabel into newly synthesized DNA. The limit of detection of DNA adducts in the glandular stomach was 1.0. In a second experiment, [7-3H]SO was administered by i.p. injection to male B6C3F1 mice. Liver DNA was analyzed after 2 h. No radioactivity was detectable at a limit of detection of CBI less than 0.6. In agreement with the relatively long half-life of SO in animals, the chemical reactivity of SO appears to be too low to result in a detectable production of DNA adducts in an in vivo situation. Upon comparison with the DNA-binding of other carcinogens, a purely genotoxic mechanism of tumorigenic action of SO is unlikely. The observed tumorigenic potency in the forestomach could be the result of strong tumor promotion by high-dose cytotoxicity followed by regenerative hyperplasia.


Multistep retrosynthesis combining a disconnection aware triple transformer loop with a route penalty score guided tree search.
Computer-aided synthesis planning (CASP) aims to automatically learn organic reactivity from literature and perform retrosynthesis of unseen molecules. CASP systems must learn reactions sufficiently precisely to propose realistic disconnections, while avoiding overfitting to leave room for diverse options, and explore possible routes such as to allow short synthetic sequences to emerge. Herein we report an open-source CASP tool proposing original solutions to both challenges. First, we use a triple transformer loop (TTL) predicting starting materials (T1), reagents (T2), and products (T3) to explore various disconnection sites defined by combining systematic, template-based, and transformer-based tagging procedures. Second, we integrate TTL into a multistep tree search algorithm (TTLA) prioritizing sequences using a route penalty score (RPScore) considering the number of steps, their confidence score, and the simplicity of all intermediates along the route. Our approach favours short synthetic routes to commercial starting materials, as exemplified by retrosynthetic analyses of recently approved drugs.


A novel SPAST gene mutation identified in a Chinese family with hereditary spastic paraplegia.
Hereditary spastic paraplegia is a heterogeneous group of clinically and genetically neurodegenerative diseases characterized by progressive gait disorder. Hereditary spastic paraplegia can be inherited in various ways, and all modes of inheritance are associated with multiple genes or loci. At present, more than 76 disease-causing loci have been identified in hereditary spastic paraplegia patients. Here, we report a novel mutation in SPAST gene associated with hereditary spastic paraplegia in a Chinese family, further enriching the hereditary spastic paraplegia spectrum. Whole genomic DNA was extracted from peripheral blood of the 15 subjects from a Chinese family using DNA Isolation Kit. The Whole Exome Sequencing of the proband was analyzed and the result was identified in the rest individuals. RaptorX prediction tool and Protein Variation Effect Analyzer were used to predict the effects of the mutation on protein tertiary structure and function. Spastic paraplegia has been inherited across at least four generations in this family, during which only four HSP patients were alive. The results obtained by analyzing the Whole Exome Sequencing of the proband exhibited a novel disease-associated in-frame deletion in the SPAST gene, and this mutation also existed in the rest three HSP patients in this family. This in-frame deletion consists of three nucleotides deletion (c.1710_1712delGAA) within the exon 16, resulting in lysine deficiency at the position 570 of the protein (p.K570del). This novel mutation was also predicted to result in the synthesis of misfolded SPAST protein and have the deleterious effect on the function of SPAST protein. In this case, we reported a novel mutation in the known SPAST gene that segregated with HSP disease, which can be inherited in each generation. Simultaneously, this novel discovery significantly enriches the mutation spectrum, which provides an opportunity for further investigation of genetic pathogenesis of HSP.


Decreased Cannabinoid CB1 Receptors in Male Tobacco Smokers Examined With Positron Emission Tomography.
Previous studies showed reduction of brain cannabinoid CB1 receptors in adults with cannabis and alcohol use disorders. Preclinical data suggest that these receptors also contribute to nicotine reward and dependence. Tobacco smoking may confound clinical studies of psychiatric disorders because many patients with such disorders smoke tobacco. Whether human subjects who smoke tobacco but are otherwise healthy have altered CB1 receptor binding in brain is unknown. We measured CB1 receptors in brains of 18 healthy men who smoke tobacco (frequent chronic cigarette smokers), and 28 healthy men who do not smoke tobacco, using positron emission tomography and [18F]FMPEP-d2, a radioligand for CB1 receptors. We collected arterial blood samples during scanning to calculate the distribution volume (VT), which is nearly proportional to CB1 receptor density. Repeated-measures analysis of variance compared VT between groups in various brain regions. Brain CB1 receptor VT was about 20% lower in subjects who smoke tobacco than in subjects who do not. Decreased VT was found in all brain regions, but reduction did not correlate with years of smoking, number of cigarettes smoked per day, or measures of nicotine dependence. Tobacco-smoking healthy men have a widespread reduction of CB1 receptor density in brain. Reduction of CB1 receptors appears to be a common feature of substance use disorders. Future clinical studies on the CB1 receptor should control for tobacco smoking.


Nonopsonic phagocytosis of nonmucoid Pseudomonas aeruginosa by human neutrophils and monocyte-derived macrophages is correlated with bacterial piliation and hydrophobicity.
We have shown previously that some strains of Pseudomonas aeruginosa from patients with cystic fibrosis are phagocytized by human polymorphonuclear leukocytes in the absence of serum opsonins. The purpose of this study was to identify the bacterial features which render certain strains susceptible to nonopsonic phagocytosis. Three strains were phagocytized by human neutrophils and monocyte-derived macrophages, and two were not, as determined by luminol-enhanced chemiluminescence, visual inspection of stained smears, and bactericidal assay. Strains that were phagocytized formed pellicles when grown in static broth, but the phagocytosis-resistant strains did not. The phagocytosis-susceptible strains were more heavily piliated and more hydrophobic than the resistant strains. Bacteria exposed to heat (60 degrees C) or UV irradiation were depiliated, as assessed by electron microscopy, and rendered resistant to phagocytosis. When P. aeruginosa was grown on agar, it was piliated, hydrophobic, and susceptible to nonopsonic phagocytosis, but when grown to stationary phase in shaken broth, it was nonpiliated, less hydrophobic, and resistant to phagocytosis. It appears that nonopsonic phagocytosis of certain P. aeruginosa strains by human polymorphonuclear leukocytes and macrophages is facilitated by hydrophobic interactions which may be determined in part by pili.


Lateral Meniscal Allograft Transplant via a Medial Approach Leads to Less Extrusion.
Accurate positioning of the bony bridge is crucial to prevent extrusion of meniscal allografts after transplant. However, oblique or lateralized placement of the bony bridge of the lateral meniscal allograft may occur due to technical error or a limited visual field. The patellar tendon may be an obstacle to approaching the anterior horn of the lateral meniscus, resulting in a laterally placed allograft. Therefore, lateral meniscal transplant through a medial arthrotomy would be an alternative approach. However, no report exists regarding allograft extrusion when comparing medial and lateral arthrotomy techniques in lateral meniscal transplants. Extrusion of the midbody of the allograft is less severe and the rotation of the bony bridge is less oblique in lateral meniscal allograft transplants through the medial parapatellar approach than those through the lateral approach. Cohort study; Level of evidence, 3. A bony bridge was used to perform 55 lateral meniscal transplants through either a medial or a lateral arthrotomy. Thirty-two allografts were transplanted through a medial arthrotomy and 23 were transplanted through a lateral arthrotomy, not randomly. Because correct positioning of the bony trough through the medial arthrotomy was easier than that through the lateral arthrotomy, the method of the arthrotomy was changed for the latter. The procedure for both groups was identical except for the arthrotomy technique, and rehabilitation was identical for both groups. Follow-up magnetic resonance imaging was conducted for all patients to measure the postoperative extrusion and obliquity of the bony bridge of the allograft. On the coronal view, extrusion was measured as the distance between the outer edge of the articular cartilage of the lateral tibial plateau and the outer edge of the meniscal allograft. On the axial view, a line (line B) was drawn along the longitudinal axis of the bony bridge. The posterior tibial condylar tangential line was drawn between the medial and lateral posterior tibial condylar cortices. A line (line T) was drawn perpendicular to the posterior tibial condylar tangential line. The angle (trough angle) between lines B and T was measured. Postoperative extrusion and the trough angle were compared between the medial and lateral arthrotomy groups. The median extrusion of the midbody of the allograft was 2.2 mm (interquartile range [IQR], 2.4 mm; range, 0-4.6 mm) in the medial arthrotomy group and 3.1 mm (IQR, 1.5 mm; range, 0-5.3 mm) in the lateral arthrotomy group ( P = .001). Seven (21.9%) patients demonstrated extrusion in the medial arthrotomy group, and 15 (65.2%) patients had extrusion in the lateral group ( P = .002). The median trough angle was 0.9° (IQR, 9.3°; range, -8.8-15.8°) in the medial arthrotomy group and 11.6° (IQR, 2.8°; range, 3-19.8°) in the lateral arthrotomy group ( P < .001). Based on this experience, lateral meniscal allograft transplant through a medial arthrotomy is preferred to decrease postoperative extrusion of the allograft.


Phenyl Esters Are Potent Inhibitors of Caseinolytic Protease P and Reveal a Stereogenic Switch for Deoligomerization.
Caseinolytic protease P (ClpP) represents a central bacterial degradation machinery that is involved in cell homeostasis and pathogenicity. The functional role of ClpP has been studied by genetic knockouts and through the use of beta-lactones, which remain the only specific inhibitors of ClpP discovered to date. Beta-lactones have served as chemical tools to manipulate ClpP in several organisms; however, their potency, selectivity and stability is limited. Despite detailed structural insights into the composition and conformational flexibility of the ClpP active site, no rational efforts to design specific non-beta-lactone inhibitors have been reported to date. In this work, an unbiased screen of more than 137 000 compounds was used to identify five phenyl ester compounds as highly potent ClpP inhibitors that were selective for bacterial, but not human ClpP. The potency of phenyl esters largely exceeded that of beta-lactones in ClpP peptidase and protease inhibition assays and displayed unique target selectivity in living S. aureus cells. Analytical studies revealed that while phenyl esters are cleaved like native peptide substrates, they remain covalently trapped as acyl-enzyme intermediates in the active site. The synthesis of 36 derivatives and subsequent structure-activity relationship (SAR) studies provided insights into conserved structural elements that are important for inhibition potency and acylation reactivity. Moreover, the stereochemistry of a methyl-substituent at the alpha position to the ester, resembling amino acid side chains in peptide substrates, impacted ClpP complex stability, causing either dissociation into heptamers or retention of the tetradecameric state. Mechanistic insights into this intriguing stereo switch and the phenyl ester binding mode were obtained by molecular docking experiments.


Molecular modeling studies of "flap up" mannosyl cation mimics.
The importance of inhibitors of glycosidases as therapeutic agents for viral, proliferative, and metabolic diseases is being increasingly recognised. Several years ago we reported that the activities of mannosidase inhibitors may be explained in terms of their similarity to the mannosyl cation intermediate postulated to form during the enzyme-catalyzed processing of oligosaccharide substrates. Recently, the validity of this model has been called in to question by some authors. We report recent molecular modeling studies undertaken to clarify this apparent contradiction. Mannostatin can indeed bind in a fashion which bears a close similarity to the mannosyl cation. Moreover we have shown that (-)-mannostatin is not able to adopt a similar binding mode to that of the mannosyl cation. As additional proof, Farr et al. have synthesized a trihydroxycyclopentylamine as a direct mimic of our mannosyl cation model. Satisfyingly, this compound shows potent inhibition of Jack Bean alpha-mannosidase, as predicted by the model. The inactivity of aminotrihydroxyhexahydro-1H-azepine against mannosidases can be explained in terms of the relative energies of the axial versus equatorial conformations of the critical hexahydroazepine ring substituents.


Monitoring the synthetic reaction of a polyamide/peptide conjugate using electrospray ionization mass spectrometry.
Recently, the chemical structures of a series of monoimidazole/polyamine conjugates were studied in this laboratory using electrospray ionization mass spectrometry (ESI-MS) combined with tandem mass spectrometry (ESI-MS/MS). The method was found to be a powerful tool for the identification of this class of compounds. During the synthesis of targeted polyamide/peptide conjugates as derivatives or analogues of netropsin and distamycin, the method was applied to analyze and track the coupling reaction for the formation of the polyamide, which was difficult to achieve using thin layer chromatography (TLC). Characteristic fragmentation pathways for a nitro-monoimidazole conjugate, an amino-monoimidazole conjugate, and the final product (a nitro-diimidazole conjugate) were explored. The fragmentations of these conjugates were strongly affected by the presence of an amino group instead of a nitro group in the molecule, and led to the identification of the three compounds in the reacting solution or in the final reaction mixture. Consequently, the reaction could be monitored successfully and the synthetic route optimized.


Effects of temperature on actin polymerized by Ca2+. Direct evidence of fragmentation.
When the temperature is lowered from 20 to 4 degrees C, the specific viscosity of actin polymerized in the presence of either 4 mM-CaCl2 or 2 mM-MgCl2, but not of actin polymerized in the presence of 90 mM-KCl, is decreased by 50% in the absence of free ATP. Addition of ATP restores the viscosity of the actin polymerized by Mg2+, but not that of actin polymerized by Ca2+, to the original value. The effect of temperature on actin polymerized in the presence of Ca2+ is due to (a) polymer-into-monomer conversion, (b) latero-lateral aggregation of filaments, and (c) fragmentation of the filaments. Fragmentation, as demonstrated by fractional centrifugation and electron microscopy, was the most important of these.


Quantitative, multiplexed workflow for deep analysis of human blood plasma and biomarker discovery by mass spectrometry.
Proteomic characterization of blood plasma is of central importance to clinical proteomics and particularly to biomarker discovery studies. The vast dynamic range and high complexity of the plasma proteome have, however, proven to be serious challenges and have often led to unacceptable tradeoffs between depth of coverage and sample throughput. We present an optimized sample-processing pipeline for analysis of the human plasma proteome that provides greatly increased depth of detection, improved quantitative precision and much higher sample analysis throughput as compared with prior methods. The process includes abundant protein depletion, isobaric labeling at the peptide level for multiplexed relative quantification and ultra-high-performance liquid chromatography coupled to accurate-mass, high-resolution tandem mass spectrometry analysis of peptides fractionated off-line by basic pH reversed-phase (bRP) chromatography. The overall reproducibility of the process, including immunoaffinity depletion, is high, with a process replicate coefficient of variation (CV) of <12%. Using isobaric tags for relative and absolute quantitation (iTRAQ) 4-plex, >4,500 proteins are detected and quantified per patient sample on average, with two or more peptides per protein and starting from as little as 200 μl of plasma. The approach can be multiplexed up to 10-plex using tandem mass tags (TMT) reagents, further increasing throughput, albeit with some decrease in the number of proteins quantified. In addition, we provide a rapid protocol for analysis of nonfractionated depleted plasma samples analyzed in 10-plex. This provides ∼600 quantified proteins for each of the ten samples in ∼5 h of instrument time.


Polymerizable 2-Propionic-3-methylmaleic Anhydrides as a Macromolecular Carrier Platform for pH-Responsive Immunodrug Delivery.
The design of functional polymers coupled with stimuli-triggered drug release mechanisms is a promising achievement to overcome various biological barriers. pH trigger methods yield significant potential for controlled targeting and release of therapeutics due to their simplicity and relevance, especially upon cell internalization. Here, we introduce reactive polymers that conjugate primary or secondary amines and release potential drugs under acidic conditions. For that purpose, we introduced methacrylamide-based monomers with pendant 2-propionic-3-methylmaleic anhydride groups. Such groups allow the conjugation of primary and secondary amines but are resistant to radical polymerization conditions. We, therefore, polymerized 2-propionic-3-methylmaleic anhydride amide-based methacrylates via reversible addition-fragmentation chain transfer (RAFT) polymerization. Their amine-reactive anhydrides could sequentially be derivatized by primary or secondary amines into hydrophilic polymers. Acidic pH-triggered drug release from the polymeric systems was fine-tuned by comparing different amines. Thereby, the conjugation of primary amines led to the formation of irreversible imide bonds in dimethyl sulfoxide, while secondary amines could quantitatively be released upon acidification. In vitro, this installed pH-responsiveness can contribute to an effective release of conjugated immune stimulatory drugs under endosomal pH conditions. Interestingly, the amine-modified polymers generally showed no toxicity and a high cellular uptake. Furthermore, secondary amine-modified immune stimulatory drugs conjugated to the polymers yielded better receptor activity and immune cell maturation than their primary amine derivatives due to their pH-sensitive drug release mechanism. Consequently, 2-propionic-3-methylmaleic anhydride-based polymers can be considered as a versatile platform for pH-triggered delivery of various (immuno)drugs, thus enabling new strategies in macromolecule-assisted immunotherapy.


What explains gray matter atrophy in long-standing multiple sclerosis?
To identify the measures of focal and diffuse white matter (WM) abnormalities that are related to whole-brain, deep, and cortical gray matter (GM) atrophy in long-standing multiple sclerosis (MS). The institutional review board approved the study; all subjects gave written informed consent. Magnetic resonance (MR) imaging was performed at 3 T in 208 patients with MS of long-standing duration (disease duration ≥ 10 years) and in 60 healthy control subjects. Normalized GM volume (NGMV), normalized WM volume (NWMV), normalized deep GM volume (NDGMV), cortical thickness, and normalized lesion volume (NLV) were quantified. Tissue integrity of normal-appearing WM (NAWM) and lesions was measured by using diffusion-tensor MR imaging. Multivariate associations between measures of GM atrophy and WM abnormalities were assessed in the patient group by using multiple linear regression. NGMV, NDGMV, and cortical thickness were reduced in patients with MS (all P < .001). The final model for NGMV consisted of NWMV, NLV, and patient age and sex (adjusted R(2) = 0.58, P < .001). NWMV, NLV, and patient sex were the explanatory variables for NDGMV (adjusted R(2) = 0.75, P < .001). The model for cortical thickness consisted of fractional anisotropy of NAWM, NLV, and patient age and sex (adjusted R(2) = 0.32, P < .001). The relationship between GM atrophy and WM abnormalities was weaker in primary and secondary progressive disease than in relapsing-remitting disease. Whole-brain and deep GM atrophy were particularly explained by WM atrophy and lesion volume, while cortical atrophy was associated with NAWM integrity loss. The weaker relationship between GM atrophy and WM abnormalities in patients with progressive disease might indicate a more independent neurodegenerative disease process in these patients.


Colistin Resistance among Enterobacteriaceae Isolated from Clinical Samples in Gaza Strip.
Bacterial infections, especially drug-resistant infections, are a major global health issue. The emergence of multidrug-resistant (MDR) strains of Enterobacteriaceae and the lack of new antibiotics have worrisome prospects for all of humanity. Colistin is considered the last-line drug for MDR Gram-negative bacteria (GNB), and it is often used for treatment of respiratory infections caused by MDR-GNB. In recent years, there has been a marked increase in the incidence of colistin-resistant infections. The main objective of this study was to investigate the presence of colistin resistance among clinical GNB isolated from Gaza Strip hospitals. Clinical Enterobacteriaceae isolates (100) were obtained from microbiology laboratories of the hospitals of different geographical locations in Gaza Strip Governorate over a period of six months. Samples were cultured, and bacterial identification was performed by standard microbiological procedures. Enterobacteriaceae isolates were tested for their antimicrobial susceptibility by the disk diffusion method and the MIC method for colistin. Varying degrees of susceptibility were observed for the isolates against the tested antimicrobials even within members of the same antimicrobial class. Amikacin was the most effective drug (74%), followed by chloramphenicol (48%), fosfomycin, and gentamicin (45%). High resistance was recorded against trimethoprim (85%) and tetracycline (83%). Only 59% of the tested isolates were interpreted as susceptible, while 41% was classified as resistant. The highest resistance to colistin was found to be among the Proteus spp. (63.2%), followed by Serratia spp. (57.1%). The lowest resistance was observed among Klebsiella isolates (31.6%). Only 39.0% of meropenem-resistant Enterobacteriaceae was susceptible to colistin, while 45.8% of imipenem-resistant Enterobacteriaceae was susceptible to colistin. The overall resistance to colistin was high (41%) among tested clinical isolates. Furthermore, 89% was MDR. These limit and complicate treatment options for the infections caused by Enterobacteriaceae in Gaza Strip. This calls for immediate actions to control and monitor the use of antimicrobials in general and colistin in particular.


Weathering the economic storm together: Family identification predicts future well-being during COVID-19 via enhanced financial resilience.
The economic crisis precipitated by the coronavirus disease (COVID-19) pandemic has placed considerable financial pressures on households across the world. These are compounded by the enforced isolation accompanying pandemic restrictions, during which individuals can struggle to access external assistance and often need to rely heavily on the social, emotional, and financial support of other family members. Previous research indicates that family financial stress has negative consequences for the mental health and well-being of members, but that heightened family identification can provide individuals with a stronger sense of collective financial resilience. In the present study, an online longitudinal survey of U.K. residents (N = 172) shows that, in summer 2020, the positive relationship between individuals' family identification and their well-being 1 month later was mediated by levels of perceived family financial efficacy and financial stress. These findings build upon existing evidence of the pivotal role of the family in financial well-being and suggest that supporting family units to cope with shared financial challenges may have psychological benefits over and above supporting individual family members. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Cyclin-Dependent Kinases 4/6 Inhibitors in Breast Cancer: Current Status, Resistance, and Combination Strategies.
Dysregulated activation of the cyclin-dependent kinases (CDKs) 4/6, leading to uncontrolled cell division, is hallmark of cancers. Further study of the cell cycle will advance the cancer treatment. As powerful and effective drugs, inhibitors of CDK 4/6 have been widely used in clinical practice for several malignancies, particularly against breast cancers driven by the estrogen receptor (ER). Three CDK4/6 inhibitors, including palbociclib (PD0332991), ribociclib (LEE011) and abemaciclib (LY2835219), have been approved by the US Food and Drug Administration (FDA) for the treatment of hormone receptor-positive, human epidermal growth factor receptor 2-negative advanced or metastatic breast cancer. However, CDK4/6 inhibitors act downstream of many mitogenic signaling pathways, and this has implications for resistance. It is worth to note that the mechanisms of resistance are not very clear. Up to now, a small number of preclinical and clinical studies have explored potential mechanisms of CDK4/6 inhibitors resistance in breast cancer. On this basis, rational and effective combination therapy is under development. Here we review the current knowledge about the mechanisms and efficacy of CDK4/6 inhibitors, and summarize data on resistance mechanisms to make future combination therapies more accurate and reasonable.


EPIC modeling of soil organic carbon sequestration in croplands of Iowa.
Depending on management, soil organic carbon (SOC) is a potential source or sink for atmospheric CO(2). We used the EPIC model to study impacts of soil and crop management on SOC in corn (Zea mays L.) and soybean (Glycine max L. Merr.) croplands of Iowa. The National Agricultural Statistics Service crops classification maps were used to identify corn-soybean areas. Soil properties were obtained from a combination of SSURGO and STATSGO databases. Daily weather variables were obtained from first order meteorological stations in Iowa and neighboring states. Data on crop management, fertilizer application and tillage were obtained from publicly available databases maintained by the NRCS, USDA-Economic Research Service (ERS), and Conservation Technology Information Center. The EPIC model accurately simulated state averages of crop yields during 1970-2005 (R(2) = 0.87). Simulated SOC explained 75% of the variation in measured SOC. With current trends in conservation tillage adoption, total stock of SOC (0-20 cm) is predicted to reach 506 Tg by 2019, representing an increase of 28 Tg with respect to 1980. In contrast, when the whole soil profile was considered, EPIC estimated a decrease of SOC stocks with time, from 1835 Tg in 1980 to 1771 Tg in 2019. Hence, soil depth considered for calculations is an important factor that needs further investigation. Soil organic C sequestration rates (0-20 cm) were estimated at 0.50 to 0.63 Mg ha(-1) yr(-1) depending on climate and soil conditions. Overall, combining land use maps with EPIC proved valid for predicting impacts of management practices on SOC. However, more data on spatial and temporal variation in SOC are needed to improve model calibration and validation.


What are the health impacts of nicotine and tobacco products on young people?
Narrative review. What are the implications of new nicotine and tobacco products on adolescent health? E-cigarettes and oral nicotine products such as pouches, lozenges, tablets, gum, and gummies. These products can be addictive and can cause respiratory, cardiovascular and oral potential health problems. They also have effects on brain development. Products are attractive to adolescents, with more than 1:10 American adolescents uses them. The use of nicotine and tobacco products by adolescents puts their health at risk and may, in some instances, lead to nicotine addiction. Those providing healthcare for adolescents have an opportunity to provide advice and signpost people to resources to help them stop using such products. There is also a need for legislation to restrict the sale of e-cigarettes and tobacco products.


Development of the Inhibitors that Target the PD-1/PD-L1 Interaction-A Brief Look at Progress on Small Molecules, Peptides and Macrocycles.
Cancer immunotherapy based on antibodies targeting the immune checkpoint PD-1/PD-L1 pathway has seen unprecedented clinical responses and constitutes the new paradigm in cancer therapy. The antibody-based immunotherapies have several limitations such as high production cost of the antibodies or their long half-life. Small-molecule inhibitors of the PD-1/PD-L1 interaction have been highly anticipated as a promising alternative or complementary therapeutic to the monoclonal antibodies (mAbs). Currently, the field of developing anti-PD-1/PD-L1 small-molecule inhibitors is intensively explored. In this paper, we review anti-PD-1/PD-L1 small-molecule and peptide-based inhibitors and discuss recent structural and preclinical/clinical aspects of their development. Discovery of the therapeutics based on small-molecule inhibitors of the PD-1/PD-L1 interaction represents a promising but challenging perspective in cancer treatment.


Paw paw and cancer: annonaceous acetogenins from discovery to commercial products.
Extracts of paw paw ( Asimina triloba, Annonaceae) are among the most potent of the 3500 species of higher plants screened for bioactive compounds in our laboratories at Purdue University. The paw paw is a small tree native to eastern North America; its edible fruits (sometimes referred to as "Indiana Bananas") have nurtured mankind for centuries. Activity-directed fractionation of the paw paw extracts, using the brine shrimp lethality bioassay, led to the isolation and molecular characterization of over 50 unique annonaceous acetogenins. Fractionation of extracts from related species resulted in the identification of over 150 additional acetogenins. The annonaceous acetogenins are derivatives of long-chain (C32 or C34) fatty acids. They are potent inhibitors of mitochondrial (complex I) as well as cytoplasmic (anaerobic) production of adenosine triphosphate (ATP) and the related nucleotides. The powerful cytotoxicity, in vivo antitumor, pesticidal, antimalarial, anthelmintic, piscicidal, antiviral, and antimicrobial effects indicated a myriad of potentially useful applications. Commercial development of these compounds uses natural mixtures of active components, incorporated into pesticidal, topical, and dietary supplement products. Successful applications and commercial products include a shampoo, highly effective in treating infestations of head lice, fleas, and ticks; a series of pesticidal sprays, which protects host plants against a diversity of pests; and an ointment for treatment of oral herpes (HSV-1) and other skin afflictions. The extract (in capsule form) enhances a mixture of natural anthelmintics. In addition, an encapsulated extract has been effectively used by certain cancer patients as a botanical supplement product.


[Treatment and metaphylaxis of gout complicated by nephropathy and urolithiasis].
The evaluation of clinical efficacy of combined treatment and metaphylaxis in 58 patients with gout complicated by nephropathy and urolithiasis was performed. The study included 41 (71%) men and 27 (29%) women aged 44 to 88 years (mean age - 58 +/- 7 years). All patients received parenteral therapy with trometamol H, 5 -10 infusion for the course, an average of 7 infusions. For the metaphylaxis, all patients received biologically active supplement urisan 2 tablets 2 times a day during next three months against the background of drug therapy. Findings indicate a high clinical efficacy of the trometamol H in the combined treatment of patients with gout, complicated by nephropathy and urolithiasis, considering that improvement of renal function, microcirculation in the renal parenchyma, increased glomerular filtration rate, normalization of nitrogenous wastes levels, partial or complete dissolution of concretions of the kidneys, a significant decrease in the tophs size, an increase in motor activity were observed, which ultimately improves the quality of life for these patients. Metaphylaxis using urisan for 3 months on a background of traditional therapy contributed to a stable normalization of blood uric acid levels, which prevented the exacerbation of underlying disease and recurrent stone formation. These data allow to recommend reducing the dose of traditional anti-gout drugs and conducting repeated course of metaphylaxis with the urisan after 5-6 months during 3 months.


Corticosteroids and bacterial meningitis.
Despite timely and appropriate antimicrobial therapy of bacterial meningitis, neurologic sequelae still occur in some children. This also remains true for the third generation cephalosporins such as ceftriaxone and cefotaxime despite their extraordinary high bactericidal titers in the cerebrospinal fluid of children with bacterial meningitis. The most effective way to prevent neurologic damage in these patients is to prevent the infection from developing in the first place. This may be achievable in the very near future for meningitis due to Haemophilus influenzae type b, the most common organism causing meningitis in children in many parts of the world. Unfortunately, even though such means of prevention may be available or under development, ensuring that all children receive these immunizations is very difficult to accomplish. Furthermore, some forms of bacterial meningitis are not likely to be preventable by immunization or other methods and thus efforts to develop new means to minimize the neurologic damage due to meningitis remain important. One such effort involves the use of antiinflammatory agents in addition to antimicrobial therapy for the treatment of this infection. This review will focus on the role of one such antiinflammatory agent, corticosteroids, as adjunctive therapy for bacterial meningitis.


Revaluation of clinical and histological criteria for diagnosis of dysmetabolic iron overload syndrome.
To re-evaluate the diagnostic criteria of insulin resistance hepatic iron overload based on clinical, biochemical and histopathological findings. We studied 81 patients with hepatic iron overload not explained by known genetic and acquired causes. The metabolic syndrome (MS) was defined according to ATPIII criteria. Iron overload was assessed by liver biopsy. Liver histology was evaluated by Ishak's score and iron accumulation by Deugnier's score; steatosis was diagnosed when present in >or=5% of hepatocytes. According to transferrin saturation levels, we observed significant differences in the amount of hepatic iron overload and iron distribution, as well as the number of metabolic abnormalities. Using Receiving Operating Curve analysis, we found that the presence of two components of the MS differentiated two groups with a statistically significant different hepatic iron overload (P<0.0001). Patients with >or=2 metabolic alterations and steatosis had lower amount of hepatic iron, lower transferrin saturation and higher sinusoidal iron than patients with <2 MS components and absence of steatosis. In our patients, the presence of >or2 alterations of the MS and hepatic steatosis was associated with a moderate form of iron overload with a prevalent sinusoidal distribution and a normal transferrin saturation, suggesting the existence of a peculiar pathogenetic mechanism of iron accumulation. These patients may have the typical dysmetabolic iron overload syndrome. By contrast, patients with transferrin saturation>or=60% had more severe iron overload, few or no metabolic abnormalities and a hemochromatosis-like pattern of iron overload.


Epidemiology of hemorrhagic fever with renal syndrome in endemic area of the Republic of Korea, 1995-1998.
We conducted an epidemiologic study to understand temporal and spatial patterns of hemorrhagic fever with renal syndrome (HFRS) in the Republic of Korea (ROK). We estimated the incidence among civilians in endemic areas through the active surveillance system during the major epidemic periods, from September to December, between 1996 and 1998. We also estimated the prevalence among Korean military personnel from 1995 to 1998. In addition, we assessed seroprevalence, subclinical infection rate, and vaccination rates in both civilians and military personnel. The incidence in civilians ranged from 2.1 to 6.6 per 100, 000 person-months. The annual prevalence in the military personnel was 40-64 per 100, 000 military populations, and remained generally constant throughout the study period with seasonal variation. This is the prospective epidemiologic data set on HFRS in the ROK since the inactivated Hantaan virus vaccine was licensed for use in the late 1990s. These results will be invaluable in establishing a national immunization program against HFRS.


Evidence of off-label inhalation therapy on pediatric asthma practice in Japan.
In Japan, many asthma inhalers do not have formal approval for use in the pediatric population because of the lack of domestic data. In real-world settings, however, numerous off-label medications are prescribed. Currently, the nature of off-label prescriptions of asthma inhalers on pediatric patients in Japan remains unclear. Using public open-source national medical claims data, we investigated the real-world descriptive epidemiology of off-label prescriptions for asthma inhalers for pediatric patients. We obtained the number of off-label prescriptions of formulations for patients aged 0-14 years from anonymously summarized prescription data for a 7-year period starting from April 2014. The actual prescription numbers and their chronology over time were then analyzed. In 2019, 143,439 asthma inhalers were used off label in children and adolescents. Overall, 96.1% were inhaled corticosteroids (ICSs) or long-acting beta stimulants (LABAs), and 3.9% were high-dose ICS. Of ICSs and LABAs, 18.8% were off-label prescriptions. The total number of off-label ICS/LABA prescriptions and their percentage relative to the overall formulations gradually decreased but a notable disparity was observed among inhaler types. There was a surprisingly large number of off-label prescriptions of asthma inhalers in the pediatric population in Japan. The proper use of ICSs/LABAs and expansion of insurance coverage should be advocated to reduce off-label use.


Worldwide child and adolescent mental health begins with awareness: a preliminary assessment in nine countries.
To temper untoward mental health outcomes in children and adolescents, the World Psychiatric Association's Presidential Global Child Mental Health Programme, in collaboration with the WHO and the International Association of Child and Adolescent Psychiatry and Allied Professionals, established a Child Mental Health Awareness Task Force headed by Sam Tyano. Its task was to develop methodologies to increase awareness among policy-makers, community leaders, health professionals, teachers, parents, and children. Based on a prior comprehensive international search for effective techniques for information dissemination, an awareness manual was written for use by health professionals in diverse communities so as to guide the design and implementation of location specific awareness campaigns. We assessed the children, parents and teachers both before and after the campaign to determine changes in knowledge, attitudes and understanding of mental health. The school-based studies were conducted in selected communities in nine countries on five different continents distinguished by their different languages, cultures and their differing levels of economic development: Armenia, Azerbaijan, Brazil, China, Egypt, Georgia, Israel, Russia, and Uganda. In the six sites that completed all assessments, indicators of positive change in awareness of child mental health were identified, and results demonstrated an increased willingness to discuss emotional problems freely. These data support the utility of collaborating with schools so as to foster better child mental health in such under-resourced communities.


[Antifungal therapy for infants, children and adolescents with suspected or documented invasive fungal infection].
Fungal nosocomial infections have gradually and consistently increased since the 90s.This increasing threat is closely related with the growing number of people with immune system disorders and their survival. It is also related with the destruction of their physical barriers against infection due to the use of cytotoxic drugs or invasive procedures, such is the case of cancer patients and bone marrow and solid organ transplant patients. Increased survival of patients with congenital or acquired immunodeficiency, premature babies and patients with complex congenital malformations, especially in the gastrointestinal tract, also add up to this scenario. The occurrence of yeast and filamentous fungi infections, especially of the Candida species, has been on the rise. Azole agents overuse, especially fluconazole, for the treatment and prophylaxis of fungal infections has put selective pressure on Candida spp. which resulted in an increase of non-albican species such as C. krusei, C. glabrata and C. famata, among others, as well as their growing resistance to these antifungal agents.


Improvement of the physiological response of barley plants to both Zinc deficiency and toxicity by the application of calcium silicate.
An adequate availability of Zinc (Zn) is crucial for plant growth and development given the essentiality of this element. Thus, both Zn deficiency and Zn toxicity can limit crop yields. In plants, the responses to Zn imbalances involve important physiological aspects such as reactive oxygen species (ROS) accumulation, phytohormone balance, tricarboxylic acid cycle (TCA) metabolism, and organic acids (OAs) accumulation. However, a way to improve tolerance to stresses such as those produced by nutritional imbalances is the application of beneficial elements such as silicon (Si). In this study, we grew barley plants in hydroponics under Zn deficiency and toxicity conditions, applying Si in the form of CaSiO3 in order to assess its effectiveness against Zn imbalances. Parameters related to plant growth, oxidative stress, TCA enzyme activities, phytohormones and OAs accumulation were analyzed. Both Zn deficiency and toxicity reduced leaf biomass, increased ROS accumulation, and affected phytohormone and OAs concentrations and TCA enzyme activities. CaSiO3 treatment was effective in counteracting these effects enhancing Zn accumulation under Zn deficient conditions and limiting its accumulation under toxic conditions. In addition, this treatment decreased ROS levels, and improved ascorbate/glutathione and phytohormonal responses, citrate synthase activity, and malate/oxalate ratio. Therefore, this study enhanced the notion of the efficacy of CaSiO3 in improving tolerance to Zn imbalances.


Honey as an indicator of long-term environmental changes: MP-AES analysis coupled with 14C-based age determination of Hungarian honey samples.
Several studies show that the elemental content of honey entirely depends on the botanical and geographical origin, but the information is incomplete regarding its time-dependent composition changes. Twenty-six acacia and three honey samples with unknown botanical origin were collected between 1958 and 2018 and analysed for elemental composition by Microwave Plasma Atomic Emission Spectrometry (MP-AES). The elemental analysis was coupled with independent dating method by Accelerator Mass Spectrometry (AMS) to confirm the calendar age of the honey samples and test the possibility of radiocarbon based dating of bee products, which has not been applied before. According to the analytical measurements and statistical analysis, we can conclude that the elemental composition shows change with time in the acacia honey during the last five decades. We have proven that honey preserves carbon isotopic and elemental information of its production time and thus can be applied as an environmental indicator (e.g. trace urban pollutants, precipitation, local industrial or agricultural emission) in reconstruction studies by analysing the non-degradable mineral content. Our results further show that acacia honey is a suitable material for radiocarbon dating, proved by the results compared to the atmospheric radiocarbon bomb-peak. The new approach presented for investigation of honey by radiocarbon-based age determination coupled with elemental analysis can be used in biological, dietary, archaeological or other multidisciplinary studies as well. Some samples show slightly depleted radiocarbon content. This could be an indication of local fossil CO2 emission. Based on these depleted 14C results, honey could be used for atmospheric monitoring of fossil CO2 urban or industrial hot-spots.


Indium Tungsten Oxide Thin Films for Flexible High-Performance Transistors and Neuromorphic Electronics.
Thin-film transistors (TFTs) with high electrical performances (mobility > 10 cm2/V s, Vth < 1 V, SS < 1 V/decade, on/off ratio ≈ 106) obtained from the silicon- and oxide-based single-crystalline semiconductor materials require high processing temperature and hence are not suitable for flexible electronics. Amorphous oxide-based transparent electronic devices are attractive to meet emerging technological demands where crystalline oxide-/silicon-based architectures cannot provide a solution. Here, we tackle this problem by using a novel amorphous oxide semiconducting material-namely, indium tungsten oxide (IWO)-as the active channel in flexible TFTs (FTFTs). Post-annealing temperature as low as 270 °C for amorphous IWO thin films deposited by radio frequency sputtering at room temperature could result in smooth morphology ( Rrms ≈ 0.42 nm), good adhesion, and high carrier density ( n ≈ 7.19 × 1018 cm-3). Excellent TFT characteristics of flexible devices could be achieved with linear field effect mobility μFE ≈ 25.86 cm2/V s, subthreshold swing SS ≈ 0.30 V/decade, threshold voltage Vth ≈ -1.5 V, and on/off ratio Ion/ Ioff ≈ 5.6 × 105 at 3 V and stable operation during bending of the FTFT. Additionally, IWO TFTs were implemented as synapses, the building block for neuromorphic computing. Paired-pulse facilitation up to 138% was observed and showed an exponential decay resembling chemical synapses. Utilizing this characteristic, a high-pass dynamic temporal filter was devised providing increased gain from 1.55 to 21 when frequency was raised from 22 to 62 Hz. The high performance and stability of flexible TFTs obtained with IWO films demonstrate their promise for low-voltage electronic applications.


The use of a blood conservation device to reduce red blood cell transfusion requirements: a before and after study.
Anaemia and the associated need for packed red blood cell (PRBC) transfusions are common in patients admitted to the intensive care unit (ICU). Among many causes, blood losses from repeated diagnostic tests are contributory. This is a before and after study in a medical ICU of a university hospital. We used a closed blood conservation device (Venous Arterial blood Management Protection, VAMP, Edwards Lifesciences, Irvine, CA, USA) to decrease PRBC transfusion requirements. We included all adult (> or =18 years) patients admitted to the ICU with indwelling arterial catheters, who were expected to stay more than 24 hours and were not admitted for active gastrointestinal or any other bleeding. We collected data for six months without VAMP (control group) immediately followed by nine months (active group) with VAMP. A restrictive transfusion strategy in which clinicians were strongly discouraged from any routine transfusions when haemoglobin (Hb) levels were above 7.5 g/dL was adopted during both periods. Eighty (mean age 61.6 years, 49 male) and 170 patients (mean age 60.5 years, 101 male) were included in the control and active groups respectively. The groups were comparable for age, gender, Acute Physiology and Chronic Health Evaluation (APACHE) II score, need for renal replacement therapy, length of stay, and Hb levels on discharge and at transfusion. The control group had higher Hb levels on admission (12.4 +/- 2.5 vs. 11.58 +/- 2.8 gm/dL, P = 0.02). Use of a blood conservation device was significantly associated with decreased requirements for PRBC transfusion (control group 0.131 unit vs. active group 0.068 unit PRBC/patient/day, P = 0.02) on multiple linear regression analysis. The control group also had a greater decline in Hb levels (2.13 +/- 2.32 vs. 1.44 +/- 2.08 gm/dL, P = 0.02) at discharge. The use of a blood conservation device is associated with 1) reduced PRBC transfusion requirements and 2) a smaller decrease in Hb levels in the ICU.


Optimization of the Promega PowerSeq™ Auto/Y system for efficient integration within a forensic DNA laboratory.
The application of massively parallel sequencing (MPS) is growing in the forensic DNA field, as forensic DNA laboratories are continuously seeking methods to gain information from a limited or degraded forensic sample. However, the laborious nature of current MPS methodologies required for successful library preparation and sequencing leave opportunities for improvement to make MPS a practical option for processing forensic casework. In this study, the Promega PowerSeq™ Auto/Y System Prototype, a MPS laboratory workflow that incorporates multiplex amplification, was selected for optimization with the objectives to introduce automation for relieving manual processing, and to reduce the number of steps recommended by the standard protocol. Successful changes in the optimized workflow included a switch from column-based PCR purification to automatable bead-based purification, adoption of the library preparation procedures by a liquid handling robot platform, and removal of various time-consuming quality checks. All data in this study were found to be concordant with capillary electrophoresis (CE) data and previously-generated MPS results from this workflow. Read abundance and allele balance, metrics related to sample interpretation reliability, were not significantly different when compared to samples processed with the manufacturer's protocol. All the modifications implemented resulted in increased laboratory efficiency, reduced the protocol steps associated with risk of contamination and human error events, and decreased manual processing time by approximately 12h. These findings provide forensic DNA laboratories a more streamlined option when considering implementation of a MPS workflow.


Flexible global calibration of multiple cameras with nonoverlapping fields of view using circular targets.
Global calibration of multicamera systems is a difficult problem. The key is to find an appropriate calibration target. This paper proposes a flexible method to construct a global calibration target with circular targets. Any object that is to be measured by the system can be used as a calibration target with the help of a hand-held scanner. The calibration method does not need a special calibration target or multiple shots, which makes it flexible in applications. Circular targets pasted on the calibration target are used as features that are captured by cameras. The particle swarm optimization method is employed to correct the eccentricity error in the projection of circular targets. The eccentricity correction method does not need any prior knowledge except the cameras' intrinsic parameters. A synthetic data experiment was performed to validate the eccentricity correction method, and a physical experiment in a train wheelset inner-side distance measurement system was performed to validate the global calibration method. The measurement accuracy of the system was better than 0.1 mm, and the eccentricity correction method improved the three-dimensional reconstruction accuracy by about 0.1 mm.


E-cadherin's dark side: possible role in tumor progression.
In the context of cancer, E-cadherin has traditionally been categorized as a tumor suppressor, given its essential role in the formation of proper intercellular junctions, and its downregulation in the process of epithelial-mesenchymal transition (EMT) in epithelial tumor progression. Germline or somatic mutations in the E-cadherin gene (CDH1) or downregulation by epigenetic mechanisms have been described in a small subset of epithelial cancers. However, recent evidence also points toward a promoting role of E-cadherin in several aspects of tumor progression. This includes preserved (or increased) E-cadherin expression in microemboli of inflammatory breast carcinoma, a possible "mesenchymal to epithelial transition" (MET) in ovarian carcinoma, collective cell invasion in some epithelial cancers, a recent association of E-cadherin expression with a more aggressive brain tumor subset, as well as the intriguing possibility of E-cadherin involvement in specific signaling networks in the cytoplasm and/or nucleus. In this review we address a lesser-known, positive role for E-cadherin in cancer.


Rotigotine transdermal patch in the management of Parkinson's disease (PD) and its night-time use for PD-related sleep disorders.
Rotigotine (Neupro) is a non-ergolinic dopamine agonist available as a transdermal patch that can be applied once daily. To date, it is approved as monotherapy for the treatment of early Parkinson's disease (PD), and as adjunctive therapy to levodopa in the treatment of PD, including the advanced stage of the disease, when the efficacy of levodopa is reduced or becomes inconsistent and there are fluctuations in the therapeutic effect (end-dose, 'ON-OFF' phenomenon). The potential advantages of the rotigotine patch include immediacy of effect onset as intestinal absorption is not needed, constant drug delivery, and ease of use. This review provides an overview of several aspects of the use of the rotigotine transdermal patch, including the important issue of the management of sleep disorders in advanced PD patients, as well as the chemistry, preclinical and clinical pharmacology, efficacy, safety and tolerability of the drug. Furthermore, the rationale for the treatment of PD with the rotigotine transdermal patch is discussed, focusing in particular on the related sleep disorders.


Intramitochondrial localization of delta-aminolaevulate synthetase and ferrochelatase in rat liver.
Intramitochondrial loci for delta-aminolaevulate synthetase and ferrochelatase, the initial and final enzymes in haem synthesis, have been found in rat liver. Two different methods of fractionation were applied to mitochondria: (a) sonication and density-gradient centrifugation; (b) treatment with digitonin and differential centrifugation. Similar results were obtained with each technique. delta-Aminolaevulate synthetase is distributed similarly to two known matrix enzymes, malate dehydrogenase and glutamate dehydrogenase. Ferrochelatase is firmly bound to the the inner mitochondrial membrane. These results are considered in terms of the regulation of haem synthesis and in relation to mitochondrial biogenesis.


ICAM-1-activated Src and eNOS signaling increase endothelial cell surface PECAM-1 adhesivity and neutrophil transmigration.
Polymorphonuclear neutrophil (PMN) extravasation requires selectin-mediated tethering, intercellular adhesion molecule-1 (ICAM-1)-dependent firm adhesion, and platelet/endothelial cell adhesion molecule 1 (PECAM-1)-mediated transendothelial migration. An important unanswered question is whether ICAM-1-activated signaling contributes to PMN transmigration mediated by PECAM-1. We tested this concept and the roles of endothelial nitric oxide synthase (eNOS) and Src activated by PMN ligation of ICAM-1 in mediating PECAM-1-dependent PMN transmigration. We observed that lung PMN infiltration in vivo induced in carrageenan-injected WT mice was significantly reduced in ICAM-1(-/-) and eNOS(-/-) mice. Crosslinking WT mouse ICAM-1 expressed in human endothelial cells (ECs), but not the phospho-defective Tyr(518)Phe ICAM-1 mutant, induced SHP-2-dependent Src Tyr530 dephosphorylation that resulted in Src activation. ICAM-1 activation also stimulated phosphorylation of Akt (p-Ser473) and eNOS (p-Ser1177), thereby increasing NO production. PMN migration across EC monolayers was abolished in cells expressing the Tyr(518)Phe ICAM-1 mutant or by pretreatment with either the Src inhibitor PP2 or eNOS inhibitor L-NAME. Importantly, phospho-ICAM-1 induction of Src signaling induced PECAM-1 Tyr686 phosphorylation and increased EC surface anti-PECAM-1 mAb-binding activity. These results collectively show that ICAM-1-activated Src and eNOS signaling sequentially induce PECAM-1-mediated PMN transendothelial migration. Both Src and eNOS inhibition may be important therapeutic targets to prevent or limit vascular inflammation.


Paradigm shifts in inpatient psychiatric care of children: Approaching child- and family-centered care.
This article describes the components of child- and family-centered care, including a review of the literature about the delivery of family-centered care and the barriers that often prevent the provision of such care. The article describes an inpatient child psychiatric unit that has implemented an approach to care that embraces these principles. The changes in structures and policies that the unit adopted are described, with specific examples cited as illustrations of the components of this care. To describe the elements of child- and familycentered care and the ways in which this model of care may be implemented on inpatient child psychiatric units. Literature review including journal reports and articles and books. Preliminary outcomes are encouraging in the decrease in use of practices such as restraint and seclusion, and physical holds. Further research examining parents' satisfaction is warranted in determining the success of such endeavors.


The effect of oleuropein from olive leaf (Olea europaea) extract on Ca²⁺ homeostasis, cytotoxicity, cell cycle distribution and ROS signaling in HepG2 human hepatoma cells.
Oleuropein, a phenolic compound found in the olive leaf (Olea europaea), has been shown to have biological activities in different models. However, the effects of oleuropein on Ca(2+) homeostasis, cytotoxicity, cell cycle distribution and ROS signaling in liver cells have not been analyzed. Oleuropein induced [Ca(2+)]i rises only in HepG2 cells but not in AML12, HA22T or HA59T cells due to the different status of 3-hydroxy-3-methylglutaryl-CoA reductase expression. In HepG2 cells, this Ca(2+) signaling response was reduced by removing extracellular Ca(2+), and was inhibited by the store-operated Ca(2+) channel blockers 2-APB and SKF96365. In Ca(2+)-free medium, pretreatment with the ER Ca(2+) pump inhibitor thapsigargin abolished oleuropein-induced [Ca(2+)]i rises. Oleuropein induced cell cycle arrest which was associated with the regulation of p53, p21, CDK1 and cyclin B1 levels. Furthermore, oleuropein elevated intracellular ROS levels but reduced GSH levels. Treatment with the intracellular Ca(2+) chelator BAPTA-AM or the antioxidant NAC partially reversed oleuropein-induced cytotoxicity. Together, in HepG2 cells, oleuropein induced [Ca(2+)]i rises by releasing Ca(2+) from the ER and causing Ca(2+) influx through store-operated Ca(2+) channels. Moreover, oleuropein induced Ca(2+)-associated cytotoxicity that involved ROS signaling and cell cycle arrest. This compound may offer a potential therapy for treatment of human hepatoma.


Optimal Fault Detection and Diagnosis in Transcriptional Circuits Using Next-Generation Sequencing.
We propose a methodology for model-based fault detection and diagnosis for stochastic Boolean dynamical systems indirectly observed through a single time series of transcriptomic measurements using Next Generation Sequencing (NGS) data. The fault detection consists of an innovations filter followed by a fault certification step, and requires no knowledge about the possible system faults. The innovations filter uses the optimal Boolean state estimator, called the Boolean Kalman Filter (BKF). In the presence of knowledge about the possible system faults, we propose an additional step of fault diagnosis based on a multiple model adaptive estimation (MMAE) method consisting of a bank of BKFs running in parallel. Performance is assessed by means of false detection and misdiagnosis rates, as well as average times until correct detection and diagnosis. The efficacy of the proposed methodology is demonstrated via numerical experiments using a p53-MDM2 negative feedback loop Boolean network with stuck-at faults that model molecular events commonly found in cancer.


[Changes in the immune reactivity of the body in cerebral arachnoiditis].
The author evaluates the dynamics of immune reactivity of the body in patients with cerebral arachnoiditis with consideration of main clinical syndrome--hypertensive-hydrocephalic, epileptiform, autonomic-vascular dystonia syndromes in different variants of the course of the process. It is shown that regression of immune insufficiency did not depend on the leading clinical syndrome. The question of including immunomodulators for correction of immune insufficiency is discussed.


Patterns of reading ability in children with autism spectrum disorder.
This study investigated reading skills in 41 children with autism spectrum disorder. Four components of reading skill were assessed: word recognition, nonword decoding, text reading accuracy and text comprehension. Overall, levels of word and nonword reading and text reading accuracy fell within average range although reading comprehension was impaired. However, there was considerable variability across the sample with performance on most tests ranging from floor to ceiling levels. Some children read accurately but showed very poor comprehension, consistent with a hyperlexia reading profile; some children were poor at reading words and nonwords whereas others were unable to decode nonwords, despite a reasonable level of word reading skill. These findings demonstrate the heterogeneous nature of reading skills in children with ASD.


Differentiation of Sialyl Linkages Using a Combination of Alkyl Esterification and Phenylhydrazine Derivatization: Application for N-Glycan Profiling in the Sera of Patients with Lung Cancer.
Alterations in oligosaccharides and types of sialic acid (SA) attachments have been associated with different pathological states. Matrix-assisted laser desorption mass spectrometry (MS) is commonly used for glycosylation studies. However, native sialylated glycans are suppressed or not detected during MS experiments. Consequently, different approaches have been employed to neutralize the negative charge of the carboxyl group. In this study, we present the advantage of phenylhydrazine (PHN) labeling for the detection and efficient discrimination of SA linkages when this derivatization follows alkyl esterification. As expected, PHN-labeled sialylated oligosaccharides with the 2,6-linkage type can be easily recognized according to the additional shift in mass corresponding to the presence of a methyl or ethyl group. Surprisingly, oligosaccharides with the 2,3-linked SA residue instead of a lactone were detected carrying the second PHN unit. This was beneficial as no further processing after esterification was needed to stabilize the lactone form. Moreover, during tandem mass experiments, all modified glycans produced favorable fragmentation patterns with a coherent recognition of SA linkages. Although both types of esterification, herein called the EST-PHN approach, provided comparable results, methylation exhibited marginally higher linkage specificity than ethyl esterification. The simplicity and effectiveness of the methodology are demonstrated on the model compound, sialyllactose, and its applicability for biological studies is presented on N-glycan profiling in the sera of lung cancer patients.


Malignant mixed epithelial and stromal tumour of the kidney: a case report and a literature review.
Mixed epithelial and stromal tumour of the kidney (MEST) is a rare and distinctive neoplasm accounting for 0.2% of all renal cancers. Most of these tumours behave in a benign fashion but 13 cases with malignant transformation have already been reported. We present the first case of an extremely aggressive MEST with rapid recurrence after radical treatment, demonstrating objective response to chemotherapy. A 31-year-old female presented to the hospital complaining of gross hematuria. Computed tomography (CT) revealed an intraparenchymal mass in the left kidney forming a tumour thrombus in the inferior vena cava (IVC). Metastatic disease was ruled out and, under the clinical diagnosis of renal cell carcinoma, left radical nephrectomy with IVC thrombectomy was performed. The histopathological examination confirmed malignant MEST of the kidney. At the follow-up 12 months after surgery, a recurrent tumour in the left paravertebral area and a tumour thrombus in the IVC were detected. A second surgery was recommended and the mass from the paravertebral area was removed, so resection of the IVC with prosthetic replacement was performed. The histopathologic examination confirmed a recurrent malignant MEST. At the follow-up three months after the second surgery disease progression was diagnosed, so chemotherapy with ifosfamide and doxorubicin was initiated. The CT scan performed 14 months after the chemotherapy confirmed a stable process of the disease with no signs of progression. A literature review and our case report confirm the existence of extremely aggressive malignant MEST that shows response to chemotherapy. However, more reports are needed to improve our understanding about the biology of the MEST to develop any recommendations on personalized therapy.


Using radiative transfer equation to model absorption by thin Cu(In,Ga)Se2 solar cells with Lambertian back reflector.
We investigate the optical absorption in a thin Cu(In,Ga)Se(2) solar cell with a Lambertian white paint beneath a transparent back contact. Although this configuration has been proposed more than 30 years ago, it turns out that rigorous simulation of Maxwell's equations demand powerful numerical calculations. This type of approach is time consuming and does not provide a physical insight in the absorption mechanisms. Here, we use the radiative transfer equation to deal with multiple scattering of the diffuse part of the light. The collimated part is treated accounting for wave effects. Our model is in good agreement with optical measurements.


Effect of altering automatic exposure control settings and quality reference mAs on radiation dose, image quality, and diagnostic efficacy in MDCT enterography of active inflammatory Crohn's disease.
The purpose of our study was to determine whether the MDCT enterography dose can be reduced by changing automatic exposure control (AEC) setting and quality reference milliampere-seconds (mAs) without altering subjective image quality or efficacy in active inflammatory Crohn's disease. This is a prospective study of 2,310 MDCT enterography procedures performed using 16- and 64-MDCT in three cohorts (original, intermediate, and final dose levels). For 16-MDCT, the original and intermediate dose level quality reference mAs was 200, and weight-based (1 pound [0.45 kg] = 1 mAs) for the final dose level. For 64-MDCT, the original dose level quality reference mAs was 260; the mAs was 220 for intermediate and weight-based for the final dose level. For the intermediate and final dose levels, AEC was changed from strong to weak increase for obese and weak to strong decrease for slim patients. Demographic data and volume CT dose index (CTDI(vol)) were analyzed. Three readers evaluated the cases for image quality and efficacy differentiating normal from active inflammatory Crohn's disease. For 16-MDCT, CTDI(vol) decreased from 12.82 to 10.14 mGy and 10.14 to 8.7 mGy between original to intermediate and intermediate to final dose levels. For 64-MDCT, the CTDI(vol) decreased from 15.72 to 11.42 mGy and 11.42 to 9.25 mGy between original to intermediate and intermediate to final dose levels. Images were rated suboptimal or nondiagnostic more often in the intermediate dose level (p < 0.05) but not in the final. There was no reduction in diagnostic efficacy as measured by area under the ROC curve (p > 0.1443 except for one comparison with one reader). Substantial dose reduction can be achieved using weight-based quality reference mAs and altering AEC settings without affecting diagnostic efficacy in active inflammatory Crohn's disease of the terminal ileum. However, subjective image quality can be compromised at these dose settings, depending on radiologist preference.


Related RNA polymerase-binding regions in human RAP30/74 and Escherichia coli sigma 70.
RAP30/74 is a heteromeric general transcription initiation factor that binds to mammalian RNA polymerase II. The RAP30 subunit contains a region that is similar in amino acid sequence to the RNA polymerase-binding domain of the Escherichia coli transcription initiation factor sigma 70 (sigma 70). Mammalian RNA polymerase II specifically protected a serine residue in the sigma 70-related region of RAP30 from phosphorylation in vitro. In addition, human RAP30/74 bound to Escherichia coli RNA polymerase and was displaced by sigma 70. These results suggest that RAP30 and sigma 70 have functionally related RNA polymerase-binding regions.


Silver-based hybrid materials from meta- or para-phosphonobenzoic acid: influence of the topology on silver release in water.
Three novel silver-based metal-organic frameworks materials, which were synthesized from either 3-phosphono or 4-phosphonobenzoic acid and silver nitrate, are reported. The novel hybrids were synthesized under hydrothermal conditions; the pH of the reaction media was controlled by adding different quantities of urea thereby producing different topologies. Compound 1 (Ag3(4-PO3-C6H4-COO)), synthesized in the presence of urea, exhibits a compact 3D structure in which both phosphonic acid and carboxylic acid functional groups are linked to the silver-based inorganic network. Compound 2 (Ag(4-PO3H-C6H4-COOH)), which was synthesized at lower pH (without urea), has a layered structure in which only the phosphonic acid functional groups from 4-phosphonobenzoic acid moieties are linked to the silver inorganic network; the carboxylic acid groups being engaged in hydrogen bonds. Finally, material 3 (Ag(3-PO3H-C6H4-COOH)) was synthesized from 3-phosphonobenzoic acid and silver nitrate without urea. This material 3 features a layered structure exhibiting carboxylic acid functional groups linked via hydrogen bonds in the interlayer space. After the full characterization of these materials (single X-ray structure, IR, TGA), their ability to release silver salts in aqueous environment was measured. Silver release, determined in aqueous solution by cathodic stripping voltammetry, shows that the silver release capacity of these materials is dependent on the topology of the hybrids. The more compact structure 1 is extremely stable in water with only trace levels of silver ions being detected. On the other hand, compounds 2 and 3, in which only the phosphonic acid functional groups were bonded to the inorganic network, released larger quantities of silver ions into aqueous solution. These results which were compared with the silver release of the previously described compound 4 (Ag6(3-PO3-C6H4-COO)2). The results clearly show that the release capacity of silver-based metal organic framework can be tuned by modifying their topology which, in the present study, is governed by the regio-isomer of the organic precursor and the synthetic conditions under which the hybrids are prepared.


Seeking a second opinion: do patients need a second opinion when practice guidelines exist?
Patients often search for a second opinion (i.e., a search for additional information on the diagnosis and/or treatment options and the potential prognosis, which will help the patient decide what to do or not to do, where, with whom and how). The scope of this phenomenon is not well documented. Also it is not clear if this is warranted or not. This paper aims to explore whether knowing that his clinician follows practice guidelines eliminates the need of a patient's to seek a second opinion. Given that practice guidelines should allow each patient to benefit from the best current clinical evidence, one might wonder if in such a context a second opinion is still necessary, and if so, for what reasons? We review the literature to find the reasons that may prompt a patient to seek a second opinion before the implementation of practice guidelines. We then analyse, from a conceptual point of view, whether for 'informed patient' (i.e., one who knows about and understands practice guidelines) these reasons still hold after the implementation of practice guidelines and if new reasons for seeking a second opinion have emerged. We also discuss practical limitations to searching for a second opinion. We show that even if some reasons, like the search for a second opinion to check whether the treatment prescribed by the first physician is appropriate or not, can be questioned after the implementation of practice guidelines, an 'informed patient' may still wish to seek a second opinion stemming from new reasons which have emerged after the introduction of practice guidelines, e.g., to determine which practice guidelines his physician follows and whether they are appropriate to his case. We conclude that the implementation of practice guidelines will not eliminate the need for a second opinion consultation. On the contrary, the use of guidelines can even stimulate a broader request for second opinions. This conclusion however needs to be validated in an empirical study.


Study on Oral Health-Related Quality of Life in Patients After Dental Implant Treatment with Patient-Reported Outcome.
The objective of this study was to use patient-reported outcome to explore patient characteristics that influence the oral health-related quality of life of post-implant patients. The study design was a nested case-control study. The target population was patients who had undergone implant treatment at the Department of Oral Implantology, Osaka Dental University, with a superstructure fitted ≥ 3 months previously. The survey period was from April 2014 to the end of October 2016. Oral health-related quality of life was evaluated using the General Oral Health Assessment Index (GOHAI). The relationship between patient characteristics and oral health-related quality of life was investigated by binomial logistic regression analysis. The Japanese GOHAI norm for the 50s and 60s age groups was used as the dependent variable for comparison with this study population. Explanatory variables were age, sex, time of survey, implant site, type of superstructure, pre-implant number of molar occlusal support zones, pre-implant deficit status, and number of missing teeth. Of the total of 1,967 subjects, data were missing for 176 subjects, the time of survey was < 3 months since fitting of the superstructure for 1,021 subjects, and 123 subjects did not meet the inclusion criteria. Therefore, 647 subjects were finally analyzed. As a result of implant treatment, 64.5% of subjects exceeded the Japanese GOHAI norm for the same age group (52.2). From logistic regression analysis, patient characteristics that were extracted as influencing oral health-related quality of life were age < 65 years (odds ratio: 1.8); women (odds ratio: 1.8); and 1 pre-implant molar occlusal support zone (reference value of 4 zones) (odds ratio: 2.5). This study showed that implant treatment is an effective method for dental prosthetic treatment from the perspective of patient-reported outcome. In addition, patient characteristics that influence the oral health-related quality of life of post-implant patients were shown to be age < 65 years, women, and one pre-implant molar occlusal support zone. This study may help to predict posttreatment oral health-related quality of life and determine treatment plans.


Surface-enhanced Raman imaging of intracellular bioreduction of chromate in Shewanella oneidensis.
This proposed research aims to use novel nanoparticle sensors and spectroscopic tools constituting surface-enhanced Raman spectroscopy (SERS) and Fluorescence Lifetime imaging (FLIM) to study intracellular chemical activities within single bioremediating microorganism. The grand challenge is to develop a mechanistic understanding of chromate reduction and localization by the remediating bacterium Shewanella oneidensis MR-1 by chemical and lifetime imaging. MR-1 has attracted wide interest from the research community because of its potential in reducing multiple chemical and metallic electron acceptors. While several biomolecular approaches to decode microbial reduction mechanisms exist, there is a considerable gap in the availability of sensor platforms to advance research from population-based studies to the single cell level. This study is one of the first attempts to incorporate SERS imaging to address this gap. First, we demonstrate that chromate-decorated nanoparticles can be taken up by cells using TEM and Fluorescence Lifetime imaging to confirm the internalization of gold nanoprobes. Second, we demonstrate the utility of a Raman chemical imaging platform to monitor chromate reduction and localization within single cells. Distinctive differences in Raman signatures of Cr(VI) and Cr(III) enabled their spatial identification within single cells from the Raman images. A comprehensive evaluation of toxicity and cellular interference experiments conducted revealed the inert nature of these probes and that they are non-toxic. Our results strongly suggest the existence of internal reductive machinery and that reduction occurs at specific sites within cells instead of at disperse reductive sites throughout the cell as previously reported. While chromate-decorated gold nanosensors used in this study provide an improved means for the tracking of specific chromate interactions within the cell and on the cell surface, we expect our single cell imaging tools to be extended to monitor the interaction of other toxic metal species.


Influence of a Structural Prior Mask on EIT Image Reconstruction.
Electrical Impedance Tomography (EIT) is a low-cost imaging method with promising results in visualizing ventilation distribution within the lungs. However, in clinical settings, the interpretability of EIT images is often limited by blurred anatomical alignment and reconstruction artifacts. Integrating structural priors into the EIT reconstruction process can enhance the interpretability of EIT images. In this contribution, we introduced a patient-specific structural prior mask into the EIT reconstruction process. Such prior mask ensures that only conductivity changes within the lung regions are reconstructed. With the aim to investigate the influence of the structural prior mask on the EIT images, we conducted numerical simulations in terms of four different ventilation status. EIT images were reconstructed with Gauss-Newton algorithm and discrete cosine transform-based EIT algorithm. We carried out quantitative analysis including the reconstruction error and figures of merit for the evaluation. The results show that the morphological structures of the lungs introduced by the prior mask are preserved in the EIT images, and the reconstruction artefacts are also limited. In conclusion, the incorporation of the structural prior mask enhances the interpretability of EIT images in clinical settings.Clinical relevance-The correct interpretation of an EIT image is crucial for a clinical diagnosis. This research demonstrates that a structural prior mask might have the potential to improve the interpretability of an EIT image, which facilitates the clinicians with a better understanding of EIT results.


Development of uniform fenofibrate-loaded biodegradable microparticle by membrane emulsification.
Fenofibrate has shown therapeutic effects on diabetic retinopathy. However, fenofibrate can be rapidly cleared from the eye after a single intravitreal injection. Here, we aim to develop fenofibrate loaded PLGA microparticles (Feno-MP) with high drug loading and sustained in vitro release up to 6 months suitable for intravitreal injection. First, orthogonal array experimental design was applied for formulation optimization. The selected formulation parameters were used to formulate Feno-MP using homogenization method and direct membrane emulsification method. Both methods generated Feno-MP with high drug loading and sustained in vitro drug release more than 140 days. Unlike the polydisperse Feno-MP prepared using homogenization method, membrane emulsification method generated Feno-MP with uniform size distribution. By controlling the membrane pore size, 1.5 µm, 8 µm and 16 µm Feno-MP were formulated and we found that larger Feno-MP demonstrated higher drug loading, more sustained drug release in vitro with less burst drug release than the smaller Feno-MP. In conclusion, we developed Feno-MP with high drug loading and sustained release profile, and elucidated that changing the particle size could have notable impacts on drug loading and release kinetics. Formulating Feno-MP with uniform size distribution by membrane emulsification method would benefit the batch-to-batch repeatability.


A methodology for specifying PET VOI's using multimodality techniques.
Volume-of-interest (VOI) extraction for radionuclide and anatomical measurements requires correct identification and delineation of the anatomical feature being studied. We have developed a toolset for specifying three-dimensional (3-D) VOI's on a multislice positron emission tomography (PET) dataset. The software is particularly suited for specifying cerebral cortex VOI's which represent a particular gyrus or deep brain structure. A registered 3-D magnetic resonance image (MRI) dataset is used to provide high-resolution anatomical information, both as oblique two-dimensional (2-D) sections and as volume renderings of a segmented cortical surface. VOI's are specified indirectly in two dimensions by drawing a stack of 2-D regions on the MRI data. The regions are tiled together to form closed triangular mesh surface models, which are subsequently transformed into the observation space of the PET scanner. Quantification by this method allows calculation of radionuclide activity in the VOI's, as well as their statistical uncertainties and correlations. The methodology for this type of analysis and validation results are presented.


Cerebellum and auditory function: an ALE meta-analysis of functional neuroimaging studies.
Over the past two decades neuroimaging data have accumulated showing that the cerebellum, traditionally viewed only as a motor structure, is also active in a wide variety of sensory and cognitive tasks. We have proposed that instead of explicit involvement in any particular motor, sensory, or cognitive task, the cerebellum performs a much more fundamental computation involving the active acquisition of sensory data. We carried out an activation likelihood estimate (ALE) meta-analysis to determine whether neuroimaging results obtained during a wide range of auditory tasks support this proposal. Specifically, we analyzed the coordinates of 231 activation foci obtained in 15 different auditory studies selected through an extensive search of the positron emission tomography (PET) and functional magnetic resonance imaging (fMRI) literature. The studies selected represent a wide variety of purely auditory tasks using highly controlled synthesized acoustic stimuli. The results clearly revealed that in addition to temporal auditory areas of cerebral cortex, specific regions in the cerebellum are activated consistently across studies regardless of the particular auditory task involved. In particular, one area in left lateral crus I area showed the greatest volume and ALE peak value among the extratemporal regions. A subanalysis was carried out that ruled out the specific association of this cerebellar cluster with attentional demand. The results are consistent with the hypothesis that the cerebellum may play a role in purely sensory auditory processing, and are discussed in light of the broader idea of the cerebellum subserving a fundamental sensory function.


Antibacterial activity of acyclic decapeptide analogs of gramicidin S.
Three acyclic decapeptide analogs of gramicidin S, although found to possess antibacterial activity, apparently have modes of action which differ from that of the naturally occurring cyclic antibiotic. In contrast to the immediate action of gramicidin S, one of the decapeptides produced complete bacteriostasis only after several cell divisions had occurred. Furthermore, mixtures of gramicidin S with either of two of the acyclic peptides were synergistic. Some implications of these findings are discussed.


Longitudinal β-Amyloid Deposition and Hippocampal Volume in Preclinical Alzheimer Disease and Suspected Non-Alzheimer Disease Pathophysiology.
Preclinical Alzheimer disease (AD) can be staged using a 2-factor model denoting the presence or absence of β-amyloid (Aβ+/-) and neurodegeneration (ND+/-). The association of these stages with longitudinal biomarker outcomes is unknown. To examine whether longitudinal Aβ accumulation and hippocampal atrophy differ based on initial preclinical staging. This longitudinal population-based cohort study used data collected at the Knight Alzheimer Disease Research Center, Washington University, St Louis, Missouri, from December 1, 2006, to June 31, 2015. Cognitively normal older adults (n = 174) were recruited from the longitudinal Adult Children Study and Healthy Aging and Senile Dementia Study at the Knight Alzheimer Disease Research Center. At baseline, all participants had magnetic resonance imaging (MRI) scans, positron emission tomography (PET) scans with carbon 11-labeled Pittsburgh Compound B (PiB), and cerebrospinal fluid assays of tau and phosphorylated tau (ptau) acquired within 12 months. Using the baseline biomarkers, individuals were classified into preclinical stage 0 (Aβ-/ND-), 1 (Aβ+/ND-), or 2+ (Aβ+/ND+) or suspected non-AD pathophysiology (SNAP; Aβ-/ND+). Subsequent longitudinal accumulation of Aβ assessed with PiB PET and loss of hippocampal volume assessed with MRI in each group. Among the 174 participants (81 men [46.6%]; 93 women [53.4%]; mean [SD] age, 65.7 [8.9] years), a proportion (14%-17%) of individuals with neurodegeneration alone (SNAP) later demonstrated Aβ+. The rates of Aβ accumulation and loss of hippocampal volume in individuals with SNAP were indistinguishable from those without any pathologic features at baseline (for Aβ accumulation: when hippocampal volume was used to define ND, t = 0.00 [P > .99]; when tau and ptau were used to define ND, t = -0.02 [P = .98]; for loss of hippocampal volume: when hippocampal volume was used to define ND, t = -1.34 [P = .18]; when tau and ptau were used to define ND, t = 0.84 [P = .40]). Later preclinical stages (stages 1 and 2+) had elevated Aβ accumulation. Using hippocampal volume to define ND, individuals with stage 1 had accelerated Aβ accumulation relative to stage 0 (t = 11.06; P < .001), stage 2+ (t = 2.10; P = .04), and SNAP (t = 9.32; P < .001), and those with stage 2+ had accelerated Aβ accumulation relative to stage 0 (t = 4.38; P < .001) and SNAP (t = 4.08; P < .001). When ND was defined using tau and ptau, individuals with stage 2+ had accelerated Aβ accumulation relative to stage 0 (t = 4.96) and SNAP (t = 4.06), and those with stage 1 had accelerated Aβ accumulation relative to stage 0 (t = 8.44) and SNAP (t = 6.61) (P < .001 for all comparisons). When ND was defined using cerebrospinal fluid biomarkers, individuals with stage 2+ had accelerated hippocampal atrophy relative to stage 0 (t = -3.41; P < .001), stage 1 (t = -2.48; P = .03), and SNAP (t = -2.26; P = .03). More advanced preclinical stages of AD have greater longitudinal Aβ accumulation. SNAP appears most likely to capture inherent individual variability in brain structure or to represent comorbid pathologic features rather than early emerging AD. Low hippocampal volumes or elevated levels of tau or ptau in isolation may not accurately represent ongoing neurodegenerative processes.


Synergistic antitumor effects of novel HDAC inhibitors and paclitaxel in vitro and in vivo.
Preclinical studies support the therapeutic potential of histone deacetylases inhibitors (HDACi) in combination with taxanes. The efficacy of combination has been mainly ascribed to a cooperative effect on microtubule stabilization following tubulin acetylation. In the present study we investigated the effect of paclitaxel in combination with two novel HDACi, ST2782 or ST3595, able to induce p53 and tubulin hyperacetylation. A synergistic effect of the paclitaxel/ST2782 (or ST3595) combination was found in wild-type p53 ovarian carcinoma cells, but not in a p53 mutant subline, in spite of a marked tubulin acetylation. Such a synergistic interaction was confirmed in additional human solid tumor cell lines harboring wild-type p53 but not in those expressing mutant or null p53. In addition, a synergistic cytotoxic effect was found when ST2782 was combined with the depolymerising agent vinorelbine. In contrast to SAHA, which was substantially less effective in sensitizing cells to paclitaxel-induced apoptosis, ST2782 prevented up-regulation of p21(WAF1/Cip1) by paclitaxel, which has a protective role in response to taxanes, and caused p53 down-regulation, acetylation and mitochondrial localization of acetylated p53. The synergistic antitumor effects of the paclitaxel/ST3595 combination were confirmed in two tumor xenograft models. Our results support the relevance of p53 modulation as a major determinant of the synergistic interaction observed between paclitaxel and novel HDACi and emphasize the therapeutic interest of this combination.


Integration and utilization of modern technologies in nephrolithiasis research.
Nephrolithiasis, or stones, is one of the oldest urological diseases, with descriptions and treatment strategies dating back to ancient times. Despite the enormous number of patients affected by stones, a surprising lack of conceptual understanding of many aspects of this disease still exists. This lack of understanding includes mechanisms of stone formation and retention, the clinical relevance of different stone compositions and that of formation patterns and associated pathological features to the overall course of the condition. Fortunately, a number of new tools are available to assist in answering such questions. New renal endoscopes enable kidney visualization in much higher definition than was previously possible, while micro-CT imaging is the optimal technique for assessment of stone microstructure and mineral composition in a nondestructive fashion. Together, these tools have the potential to provide novel insights into the aetiology of stone formation that might unlock new prevention and treatment strategies, and enable more effective management of patients with nephrolithiasis.


Time and type of prophylactic surgery for familial adenomatosis coli.
Data on 61 kinds of familial adenomatosis coli registered in Finland, including 185 affected members, were collected to evaluate the chronological evolution of carcinoma in this disease. The long-term results for 52 patients undergoing colectomy and ileorectostomy were reviewed. Colorectal carcinoma occurred in 105 patients, with cumulative percentages of 1, 6, 16, 28, and 42 at 20, 25, 30, 35, and 40 years of age. Only three out of 50 call-up patients (6%) had a carcinoma, as compared with 62% in the propositi group. No cases of rectal stump carcinomas were recorded after 52 ileorectostomies in a follow-up period of up to 21 years (mean 6 +/- 5 S.D. years). It was concluded that prophylactic colectomy must usually be performed at 20 to 25 years of age, at the latest. Despite conflicting earlier evidence, colectomy and ileorectal anastomosis may still be a viable alternative, providing that the rectal stump is very short (5 to 10 cm) and the surveillance and treatment of all new adenomas is effective.


Stability of Progranulin Under Pre-Analytical Conditions in Serum and Cerebrospinal Fluid.
Progranulin (PGRN) levels in blood and cerebrospinal fluid (CSF) are increasingly studied as potential markers for neurodegenerative disorders. We aimed to 1) characterize two commercially available PGRN ELISAs on several assay validation parameters, 2) assess the stability of PGRN in serum and CSF under pre-analytical conditions, and 3) compare stability in the two assays. Intra- and inter-assay variation, inter-lot variation, linearity, lower limit of detection, and kit correlations were assessed for the Adipogen and R&D PGRN ELISA kits. Blood and serum samples were experimentally exposed to ≤9 freeze/thaw cycles, delayed processing for ≤24 h at room temperature and 4°C, and to temperature stability tests for ≤3 weeks at -20°C, 4°C, room temperature, and 37°C. Both commercial PGRN ELISA kits showed acceptable ranges for intra- and inter-assay variation, where the R&D kit performed more accurate than the Adipogen kit, especially for inter-assay variation (intra-assay serum: 6.7 and 8.3%, respectively; inter-assay serum: 9.2 and 21.0%; intra-assay CSF: 3.6 and 12.0%; inter-assay CSF: 16.0 and 44.5%). Absolute serum PGRN concentrations were 1.9-fold higher in Adipogen than R&D (p < 0.001) and strongly correlated between both kits (ρ= 0.86, p < 0.0001) and CSF PGRN levels were on the borderline of detection in both kits. PGRN was typically stable under all pre-analytical conditions addressed, although two weeks at 37°C resulted in decreased PGRN concentrations in CSF, only when using the Adipogen kit. These results support further examination of PGRN as a potential marker in neurodegenerative diseases, since PGRN is stable in serum and CSF and can be measured using ELISA kits from several providers.


Correlation between the clinical severity of laryngomalacia and endoscopic findings.
To correlate the clinical severity of laryngomalacia (LM) with endoscopic findings, swallowing evaluations and polysomnography in a cohort of patients. Method: We conducted a retrospective analysis between 2017-2018 on a cohort of patients diagnosed with upper airway obstruction (UAO), stridor, noisy breathing or laryngomalacia. This study took place at the Pediatric Pulmonology Department, Riley Children's Hospital, Indianapolis, United States of America. There were 157 patients with laryngomalacia included in the study. Patients with severe LM were significantly younger than those with mild LM (p=0.0214) and moderate LM (p=0.0220). Subjects with type I of LM were significantly older than type III (p=0.0051). When associations were tested between polysomnogram (PSG) variables and clinical severity, there were significant associations with age at PSG. The overall apnea-hypopnea index (AHI) in mild (p=0.0103) and moderate (p=0.0242) were significantly lower than the severe group. The rapid eye movement (REM) AHI was significantly lower in moderate cases than severe (p=0.0134). The end-tidal carbon dioxide (EtCO2) peak was significantly lower in mild cases than severe (p=0.0141). The total sleep time (TST) peripheral capillary oxygen saturation (SpO2) 90% occurs in both mild (p=0.0197) and moderate (p=0.0498) were significantly lower than the severe group. The severity of the clinical manifestations of LM did not correlate with the different endoscopic types in our study. The presence of cyanosis was associated with type III LM. Rapid eye movement AHI and EtCO2 in polysomnogram were remained significantly associated with clinical severity.