Varus-valgus laxity correlates with pain in osteoarthritis of the knee.
Pain during osteoarthritis (OA) of the knee does not necessarily correlate with the severity of the radiographic grade, and the mechanism of pain has not been completely clarified. The purpose of this study was to evaluate risk factors for pain in the knee OA using epidemiologic analyses. We evaluated 518 out of 4183 people over the age of 40 (156 males and 362 females) from Shinyoshitomi village, Japan. Mean ages were 63.8 years for men and 60.7 years for women. Screening included a physical examination of the knee and a standing AP roentgenogram of the bilateral knee. Radiographic OA was defined as a Kellgren-Lawrence grade 2 or higher. All data were coded and pain risk factors were evaluated using a multiple logistic regression model. Radiographic OA was observed in 18.4% of men and 26% of women. Of these subjects with OA, 10.9% of men and 32.5% of women complained of knee pain. Seven factors-age, gender, BMI, radiographic grade, varus-valgus laxity, torque of quadriceps muscles, and varus-valgus alignment-were evaluated as potential risk factors for pain. A significant increase in the odds ratio was observed with varus-valgus laxity (p=0.005; odds ratio, 3.04). Our results suggest that varus-valgus laxity is a risk factor for pain during knee OA.


Online patient education interventions in type 2 diabetes or cardiovascular disease: A systematic review of systematic reviews.
Online patient education is a growing form of support to patients with chronic conditions, including type 2 diabetes (Type 2 DM) and cardiovascular disease (CVD). Multiple systematic reviews have been undertaken on this topic with conflicting results. We aim to explore the applications of online patient education in Type 2 DM and CVD and synthesise current evidence. A systematic review of systematic reviews was performed. Ovid Medline, EMBASE and Cochrane Database were searched between January 2005 and May 2018. Systematic reviews considering patient outcomes of online education interventions for adults with Type 2 DM and/or CVD were included. Quality assessment and data extraction was carried out in duplicate, and data combined using narrative synthesis. The PROSPERO registration number is CRD42016034018. Twenty-three systematic reviews were eligible, synthesizing evidence from 87 distinct primary studies. Six reviews were high quality, nine used meta-analysis. Biological, behavioural, psychological, knowledge and self-efficacy measures are all potential targets. The outcomes most consistently showing benefits were knowledge and social support. Online patient education has wide ranging benefits for people with Type 2 DM or CVD. Strengths of this review include its comprehensive synthesis of the large amount of literature on this topic.


Synthesis of photocaged 6-O-(2-nitrobenzyl)guanosine and 4-O-(2-nitrobenzyl) uridine triphosphates for photocontrol of the RNA transcription reaction.
6-O-(2-Nitrobenzyl)guanosine and 4-O-(2-nitrobenzyl)uridine triphosphates (NBGTP, NBUTP) were synthesized, and their biochemical and photophysical properties were evaluated. We synthesized NBUTP using the canonical triphosphate synthesis method and NBGTP from 2',3'-O-TBDMS guanosine via a triphosphate synthesis method by utilizing mild acidic desilylation conditions. Deprotection of the nitrobenzyl group in NBGTP and NBUTP proceeded within 60s by UV irradiation at 365nm. Experiments using NBGTP or NBUTP in T7-RNA transcription reactions showed that NBGTP could be useful for the photocontrol of transcription by UV irradiation.


The type-2 peroxisomal targeting signal.
The type-2 peroxisomal targeting signal (PTS2) is one of two peptide motifs destining soluble proteins for peroxisomes. This signal acts as amphiphilic α-helix exposing the side chains of all conserved residues to the same side. PTS2 motifs are recognized by a bipartite protein complex consisting of the receptor PEX7 and a co-receptor. Cargo-loaded receptor complexes are translocated across the peroxisomal membrane by a transient pore and inside peroxisomes, cargo proteins are released and processed in many, but not all species. The components of the bipartite receptor are re-exported into the cytosol by a ubiquitin-mediated and ATP-driven export mechanism. Structurally, PTS2 motifs resemble other N-terminal targeting signals, whereas the functional relation to the second peroxisomal targeting signal (PTS1) is unclear. Although only a few PTS2-carrying proteins are known in humans, subjects lacking a functional import mechanism for these proteins suffer from the severe inherited disease rhizomelic chondrodysplasia punctata.


Morphokinetic parameter comparison between embryos from couples with high or low sperm DNA fragmentation index.
To study whether time-lapse imaging can identify morphokinetic events impacted by a high sperm DNA fragmentation index (DFI). Historical cohort study. Private university-affiliated in vitro fertilization center. A total of 978 zygotes cultured until day 5 in a time-lapse imaging incubator between March 2019 and August 2020, derived from 118 patients undergoing intracytoplasmic sperm injection as a result of idiopathic male factor infertility. Kinetic markers from the point of insemination were recorded. Generalized linear mixed models adjusted for potential confounders followed by the Bonferroni post hoc test were used to compare the timing of specific events in patients with a low (<30%) or high (≥30%) sperm DFI. The recorded kinetic markers were the following: timing to pronuclei appearance and fading; timing to 2, 3, 4, 5, 6, 7, and 8 cells; and timing to start blastulation and blastulation. Timing to blastulation. Embryos derived from sperm samples with ≥30% DFI showed significantly slower divisions compared with those with <30% DFI (mean differences of 0.7 hours in timing to pronuclei appearance, 1.2 hours in timing to pronuclei fading, 1.5 hours in timing to 2 cells, 2.5 hours in timing to 3 cells, 1.8 hours in timing to 4 cells, 3.3 hours in timing to 5 cells, 3.1 hours in timing to 6 cells, 3.2 hours in timing to 7 cells, 2.7 hours in timing to 8 cells, 8.4 hours in timing to start blastulation, and 3.8 hours in timing to blastulation). The incidences of reverse or direct cleavages (9.3% vs. 4.4%; odds ratio [OR], 2.24; 95% confidence interval [CI], 1.32-3.77) and multinucleation at 2-cell (18.9% vs. 12.0%; OR, 1.70; 95% CI, 1.12-2.58) and 4-cell (14.2% vs. 6.4%; OR, 2.42; 95% CI, 1.57-3.74) stages were significantly higher in embryos deriving from ≥30% DFI than from <30% DFI. The KIDScore ranked significantly different between embryos derived from samples with <30% and ≥30% DFI. Continuous DFI was positively correlated with all timings of specific events and with the incidences of abnormal cleavage patterns (OR, 1.042; 95% CI, 1.025-1.059) and multinucleation at 2-cell stage (OR, 1.053; 95% CI, 1.030-1.076) and inversely correlated with the KIDScore rank (B, -0.218; 95% CI, -0.044 to -0.007). No significant differences were observed in clinical outcomes between the groups. Embryo morphokinetic parameters are negatively impacted by high sperm DFI, resulting in delayed cell cleavage and blastulation.


Effect of cleaning materials on microtensile bond strength of resin composite to primary dentin contaminated with root canal sealers.
There have been no studies on the bond strength of adhesives with dentin surfaces contaminated with root canal sealers in primary teeth without underlying permanent teeth germs. This study investigated the cleaning materials used for primary tooth dentin contaminated with root canal sealers. The aim was to increase the success rate of root canal treatment in pedodontics clinics and retain the teeth for longer. The occlusal enamel layer was removed, followed by the application of root canal sealers (AH Plus or MTA Fillapex) to the dentin and cleaning using different irrigation solutions (saline, NaOCl, and ethanol). The specimens were restored using a self-etch adhesive and composite. Sticks with a thickness of approximately 1 mm were obtained from each sample, and the bond strengths were measured using a microtensile testing device. The interfacial morphology of the bonded space was evaluated using scanning electron microscopy. The control and AH Plus saline groups had the highest bond strengths. The groups cleaned using ethanol had the lowest bond strengths (p < 0.01). Cleaning the dentin with saline-soaked cotton pellets provided the best bond strengths. Therefore, saline is the most effective material for removing both epoxy resin- and calcium silicate-based root canal sealers from the access cavity.


Reference values of IGF-I in children from birth to 5 years of age, in Burkina Faso, using blood samples on filter paper.
The aims of this study were to validate the use of filter paper to measure insulin-like growth factor-I (IGF-I) and to establish normal levels of IGF-I in children appearing healthy, from birth to 5 years of age in an African population. We determined IGF-I from blood collected on filter paper. We validated this method by comparing the IGF-I values from dried blood spots on filter paper (kept at 4 degrees C and ambient temperature) and from serum among 13 children under 5. IGF-I were measured by the classical IGF-I RIA, after separation of the IGF-I from its binding proteins, using Sep-Pak chromatography. To establish normal levels of IGF-I, we conducted a cross-sectional study and collected blood samples with filter paper among 360 children in Ouagadougou (Burkina Faso). IGF-I determined from dried blood spots on filter paper were in good agreement with IGF-I levels obtained from blood serum, whether the filter papers were kept at 4 degrees C or at ambient temperature. The results of IGF-I-levels in apparently healthy children showed that geometric mean IGF-I ranged from 27 microg/l in boys younger than five months to 31 microg/l in 5-year-old boys. In girls, mean IGF-I ranged from 29 microg/l for girls younger than five months to 45 microg/l at the age of 5. From birth to 24 months, IGF-I decreased by 0.32+/-0.08 microg/l/month in boys and by 0.27+/-0.06 microg/l/month in girls and these decreases were not significantly different (p=0.95). After the age of 24 months, there was an increase in IGF-I of 4.9+/-1.3 microg/l/year in boys and of 8.4+/-0.8 microg/l/year in girls. This increase was indeed significantly different (p<0.001). Reference values of IGF-I for African boys and girls were determined. They will be used for endocrine evaluations and nutritional monitoring.


Safety of intermediate left main stenosis revascularization deferral based on fractional flow reserve and intravascular ultrasound: A systematic review and meta-regression including 908 deferred left main stenosis from 12 studies.
Current guidelines recommend intravascular ultrasound (IVUS) or fractional flow reserve (FFR) to decide upon ambiguous left main (LM) disease. However, no study has compared the safety of LM revascularization deferral based on FFR or IVUS. MEDLINE/PubMed was systematically screened for studies reporting on deferred treatment of angiographically ambiguous LM based upon FFR or IVUS evaluation. Baseline, angiographic and outcome data were appraised and pooled separately for each strategy according to random-effect models with inverse-variance weighting. A total of 908 LM stenoses from 7 FFR and 5 IVUS studies were included with median follow-up of 29.0 and 31.5 months respectively. Per year of follow-up occurrence of overall MACE were 5.1% in FFR group and 6.4% in IVUS group while death, myocardial infarction, LM revascularization were respectively 2.6%, 1.5% and 1.8% vs. 3.0%, 0.5% and 2.2%. Meta-regression analysis suggested the influence of a distal LM stenosis on MACE in FFR group (β = 0.06, p = 0.01) and age in IVUS group (β = 0.4, p = 0.001). In individual studies several independent predictors of MACE were identified including use of lower doses of intracoronary adenosine (OR 1.39, p = 0.04) in FFR group and plaque burden (OR 1.34, p = 0.025), number of other diseased vessels (OR 1.39, p = 0.04) and any untreated stenosis (OR 3.80; p = 0.037) in IVUS- studies. Deferring LM intermediate stenosis on the basis of FFR or IVUS showed an acceptable and similar risk of events in a mid-term follow-up. Conversely, several different variables related to each technique showed an interaction on outcome.


The effects of the beta 2-agonist drug clenbuterol on taurine levels in heart and other tissues in the rat.
The administration of a single subcutaneous dose of clenbuterol to rats altered the level of taurine in certain tissues. Taurine levels in cardiac tissue were significantly decreased 3 h after the administration of 250 micrograms/kg of clenbuterol and remained significantly depressed at 12 h post-dose only returning to control values by 24 h. The level of taurine in the liver increased 3 h after clenbuterol administration but was lower than the control value at 24 h post dose. Lung taurine levels were significantly lower than the control value at 12 hr post dose and remained depressed until 24 h post dose. Clenbuterol caused a significant increase in taurine levels in serum and muscle at 3 and 6 hr postdosing respectively but not at other time points. Serum creatine kinase (CK), activity was slightly but significantly raised at the 12 and 24 h time point. The effects of clenbuterol on tissue taurine content were not dose-dependent over the range studied (63-500 micrograms/kg). However taurine levels in the lung were significantly reduced at all doses and in the heart were significantly lower in the treated groups at all except the lowest dose, 12 h post dosing. Liver taurine levels were significantly increased at the highest dose of 500 micrograms/kg. The reduction of taurine concentrations in the heart, caused by clenbuterol, is of concern as taurine has been shown to have protective properties in many tissues especially the heart.


Early detection but delayed appearance of a bladder tumor.
Herein is described a case of bladder cancer first detected by urinary cytology 20 years before a definite cystoscopic diagnosis could be established. To our knowledge, this represents the longest such interval on record. The case indicates the reliability of Papanicolaou studies of the urine, especially when tumor cells are identified.


The chest X-ray in pulmonary capillary haemorrhage: correlation with carbon monoxide uptake.
Serial changes in carbon monoxide uptake (KCO or DLCO/VA) were used to monitor episodes of pulmonary capillary haemorrhage in Goodpasture's syndrome (13 cases), immune complex nephritis (three cases) and idiopathic haemosiderosis (one case). Air-space shadowing on the chest X-ray (on a scoring system 0 to 12) was assessed in the light of the changes of KCO. In 14 out of 27 episodes of bleeding, the rise and fall of KCO was matched in time by the appearance and disappearance of air-space shadowing on the chest X-ray. In six episodes the chest X-ray remained normal despite a rise of KCO. In two cases air-space shadowing appeared up to 48 h after the rise of KCO. On five occasions chest X-ray abnormalities preceded the rise of KCO but chest infection or fluid overload accounted for three of these. In cases with suspected pulmonary capillary haemorrhage, measurements of carbon monoxide uptake will provide additional information and will assist in the interpretation of the chest X-ray.


Hemoglobin Abruzzo (beta143 (H21) His replaced by Arg). Consequences of altering the 2,3-diphosphoglycerate binding site.
Hemoglobin Abruzzo is an abnormal human hemoglobin with a substitution at a residue known to be involved in the binding of 2,3-diphosphoglyceric acid. It has increased oxygen affinity and reduced heme-heme interaction in the absence of organic or inorganic phosphate cofactors. In inorganic phosphate buffers the Bohr effect and heme-heme interaction are normal, but the oxygen affinity remains higher than that of hemoglobin A. CO combination in inorganic phosphate is more strongly autocatalytic than in normal hemoglobin and a slower rate of oxygen dissociation is observed. Although many of the functional differences of this variant may be attributed to the high oxygen affinity of the mutant beta chains, the interactions between subunits are also affected by the histidine to arginine substitution at beta143. Stripped hemoglobin Abruzzo appears to be significantly more dissociated than hemoglobin A. Kinetic studies indicate that interaction with organic or inorganic phosphates decreases its subunit dissociation. In all of the functional properties examined, hemoglobin Abruzzo is more sensitive to the allosteric influence of organic and inorganic anions than is hemoglobin A.


Is there any association between genetically predicted periodontitis and ischemic stroke?
Mendelian randomization study. Using a Mendelian randomization framework, the causal relationship between periodontitis [chronic periodontitis (CP) and aggressive periodontitis (AgP)] and ischemic stroke and its subtypes [cardioembolic (CE) stroke, large artery atherosclerosis (LAA), and small vessel occlusion (SVO)], were studied. Data from three large databanks, namely, UK Biobank, genome-wide association study of European ancestry, and MEGASTROKE consortium of European ancestry were used to obtain genetic variant information of CP, AgP, and ischemic stroke, respectively. UK Biobank contributed 950 cases of CP and 455,398 controls. Genome-wide association study of European ancestry contributed 851 AgP cases and 6836 controls. MEGASTROKE consortium of European ancestry contributed 34,217 cases of ischemic stroke and its subtypes and 406,111 controls. Statistical tests including inverse variant weighted method and sensitivity analyses were performed to analyse the causal inference and to verify the strength of the results. A total of 15 single nucleotide polymorphisms (SNPs) for CP was obtained as genetic instruments. No causal inference of CP on ischemic stroke was found. Among the ischemic stroke subtypes, with the exception of CE stroke, no significant causal inference of CP on LAA or SVO was found. A total of 9 single nucleotide polymorphisms (SNPs) for AgP was obtained as genetic instruments. No causal inference of AgP on ischemic stroke and its subtypes were found. Neither CP nor AgP was found to have a causal inference on ischemic stroke and most of its subtypes. A probable causal effect of CP on CE stroke was noted.


Effect of chlorpyrifos on the inhibition of the enzyme acetylcholinesterase by cross-linking in water-supply samples and milk from dairy cattle.
A methodology for the determination of chlorpyrifos in water-supply samples and in milk from dairy cattle was developed. An amperometric biosensor was used to inhibit the enzyme acetylcholinesterase (AChE), which was immobilized by the cross-linking method (crosslinks between the enzyme and the sensor). The potential applied, the amount of enzyme to be immobilized and the acetylthiocholine (ACTh) concentration were optimized before calibration and analysis of the samples was performed. The concentration of chlorpyrifos was determined in the range of 1.0×10(-6) M to 5.0×10(-2) M with a detection limit of 5.0×10(-6) M. Spiked water samples showed high recoveries (91.32% and 93.98% for low and high chlorpyrifos levels, respectively), while milk samples exhibited a matrix effect with recoveries of 82.81% and 79.77% for high and low chlorpyrifos levels, respectively. The average concentration of chlorpyrifos in the water supply samples (5.11×10(-6) M), determined using the biosensor, was compared using gas chromatography and gave an average value of 3.04×10(-6) M. The results allow it to be concluded that although chromatographic methods are still more exact, biosensors are promising tools for the determination of analytes in the field, as they have a low cost, a reduced analysis time and good reproducibility in the data.


Histone H3 and H4 tails play an important role in nucleosome phase separation.
Chromatin organization and its dynamic regulation are crucial in governing the temporal and spatial accessibility of DNA for proper gene expression. Disordered chains of nucleosomes comprise the basis of eukaryotic chromatin, forming higher-level organization across a range of length scales. Models of chromatin organization involving phase separation driven by chromatin-associating proteins have been proposed. More recently, evidence has emerged that nucleosome arrays can phase separate in the absence of other protein factors, yet questions remain regarding the molecular basis of chromatin phase separation that governs this dynamic nuclear organization. Here, we break chromatin down into its most basic subunit, the nucleosome core particle, and investigate phase separation using turbidity assays in conjunction with differential interference contrast microscopy. We show that, at physiologically-relevant concentrations, this fundamental subunit of chromatin undergoes phase separation. Individually removing the H3 and H4 tails abrogates phase separation under the same conditions. Taking a reductionist approach to investigate H3 and H4 tail peptide interactions in-trans with DNA and nucleosome core particles supports the direct involvement of these tails in chromatin phase separation. These results provide insight into fundamental mechanisms underlying phase separation of chromatin, which starts at the level of the nucleosome core particle, and support that long-range inter-nucleosomal interactions are sufficient to drive phase separation at nuclear concentrations. Additionally, our data have implications for understanding crosstalk between histone tails and provide a lens through which to interpret the effect of histone post-translational modifications and sequence variants. STATEMENT OF SIGNIFICANCE: Emerging models propose that chromatin organization is based in phase separation, however, mechanisms that drive this dynamic nuclear organization are only beginning to be understood. Previous focus has been on phase separation driven by chromatin-associating proteins, but this has recently shifted to recognize a direct role of chromatin in phase separation. Here, we take a fundamental approach in understanding chromatin phase separation and present new findings that the basic subunit of chromatin, the nucleosome core particle, undergoes phase separation under physiological concentrations of nucleosome and monovalent salt. Furthermore, the histone H3 and H4 tails are involved in phase separation in a manner independent of histone-associating proteins. These data suggest that H3 and H4 tail epigenetic factors may modulate chromatin phase separation.


Self-reported mini olfactory questionnaire (Self-MOQ): A simple and useful measurement for the screening of olfactory dysfunction.
Olfactory dysfunction is a common problem. However, too little attention has been paid to questionnaires used to evaluate quantitative olfactory dysfunction. Therefore, the current study aimed to develop a simple self-reported Mini Olfactory Questionnaire (Self-MOQ) for the screening of quantitative olfactory dysfunction in clinical practice. Two hundred and eighty-five patients who had subjective complaints of olfactory disorder participated. The Sniffin' Sticks test score was used to define functional anosmia, hyposmia, or normosmia. We assessed the factor structure as well as internal consistency, convergent validity, and discrimination performance. The results showed that the final version of the Self-MOQ included only one factor with five items. The Self-MOQ has a good internal reliability (Cronbach's α = 0.84) and validity (r = -0.60, P < 0.001). The receiver operating characteristic analyses indicated that the Self-MOQ as compared to a visual analogue scale (VAS) is an effective measure for discriminating normosmic from hyposmic/anosmic patients, anosmic patients, and hyposmic patients. The Self-MOQ is a simple, reliable and valid questionnaire to screen olfactory dysfunction in clinical practice that appears to be superior to the use of VASs but does not replace olfactory testing. 4 Laryngoscope, 2019.


The impact of pharmacy benefit managers on community pharmacy: A scoping review.
The introduction of pharmacy benefit managers (PBMs) within the United States healthcare system occurred with the aim to decrease costs and increase quality. News media and legislation have painted a picture of decreased pharmacy competition and potential negative impacts on patients and their access to affordable medications. The objective of this scoping review was to evaluate the current research literature examining the impact of PBMs on the finances of community pharmacies. Scientific journal articles published between 2010 and 2022 were included if they met the predefined objective. This scoping review identified four articles that met inclusion criteria. None of the identified articles independently quantified the financial impact of PBMs on community pharmacies. Additional research should be completed to specifically understand the financial impact on community pharmacies to help ensure the viability of community pharmacy as an integral access point for patients.


The use of remote microphone systems in unilateral hearing loss: a preliminary study among Brazilian children and teenagers.
Due to the large number of individuals with Unilateral Hearing Loss (UHL) and the recommendation to use hearing assistive devices, studies are required to define possibilities of intervention for this population. To evaluate the performance of the Remote Microphone System (RMS) in children with UHL. Prospective clinical study with a convenience sample. Eleven children (mean age of 9.2 years) with severe and profound sensorineural UHL, hearing aid users and enrolled in regular schools participated in the study. They were evaluated using the Hearing in Noise Test (HINT), the Classroom Participation Questionnaire (CPQ), and the Sustained Auditory Attention Ability Test (SAAAT) with RMS. HINT results were analyzed using variance to three criteria of repeated measures, which revealed differences between intervention, position, and time factors and significant interaction between these three factors. The comparative analysis of the results from CPQ showed significant differences in the statistical t-test (p=<0.001) for all subscales. The analysis of variance at two repeated measures criteria used in the study of SAAAT revealed a difference between intervention and time, and both interacted significantly. The RMS associated with a hearing aid was effective for individuals with UHL.


Effect of remote ischaemic conditioning on clinical outcomes in patients presenting with an ST-segment elevation myocardial infarction undergoing primary percutaneous coronary intervention.
The rationale and study design for the multicentre, randomized, controlled CONDI2/ERIC-PPCI study are discussed.


Expressions of galectin-3, glutathione S-transferase A2 and peroxiredoxin-1 by nonylphenol-incubated Caco-2 cells and reduction in transepithelial electrical resistance by nonylphenol.
Nonylphenol, an estrogenic xenobiotic widely used in the manufacture of plastics and detergents, has been found in drinking water and may therefore enter the body through the oral route. Thus, intestinal cells lining the alimentary tract serve as the body's first line of defense against this compound. In this study, the effects of nonylphenol on the human intestinal cell line Caco-2 were determined using transepithelial electrical resistance (TEER) measurement and proteomics. Results show that 10 microM nonylphenol can disrupt the tight-junction permeability of Caco-2 cells in approximately 15 min. Incubating the cells with 1 or 10 microM nonylphenol for 6 days resulted in the enhanced expressions of galectin-3 (approximately 4-fold vs. control with 1 microM; 2-fold with 10 microM), glutathione S-transferase A2 (approximately 8-fold with 1 microM; 5-fold with 10 microM) and peroxiredoxin-1 (approximately 6-fold with 1 microM; 4-fold with 10 microM). These expressions may represent a possible consortium of mechanisms by which the cells protect themselves against nonylphenol-induced stresses. To the best of our knowledge, this is the first study on the effects of nonylphenol on Caco-2 cells.


Pseudoxanthoma elasticum (PXE): ultrastructural and biochemical study on proteoglycan and proteoglycan-associated material produced by skin fibroblasts in vitro.
Pseudoxanthoma elasticum is a genetic disease characterized by progressive mineralization of elastic fibers. Previous studies suggested that other components, apart from elastin, might be involved in the alterations of this connective tissue disorder (Martinez-Hernandez and Huffer, 1974; Pasquali Ronchetti et al., 1981; 1986). Evidence is presented that proteoglycan metabolism is altered in PXE-affected patient. Urinary GAGs suggests an increased degradation of glucosamine-containing GAGs in the patient. Pulse and chase experiments on in vitro skin fibroblasts indicated a decreased rate of synthesis of [35SO4] containing GAGs or an increase of their turnover rate in PXE. Moreover, when PGs produced from skin fibroblasts were identified by ultracentrifugation and gel filtration in associative conditions, PXE fibroblasts produced a significantly higher amount of the high molecular weight fraction of sulfated PGs. This high molecular weight material was present both in the medium and in the matrix and disappeared under dissociative conditions or after treatment with hyaluronidase or with pancreas elastase. By electron microscopy, PXE fibroblasts appeared to produce and secrete an enormous amount of toluidine blue 0 positive material organized as filaments and amorphous masses. These data are in agreement with previous observations of the presence of abnormal masses of microfilaments, in the dermis of PXE patients, which were sensitive to hyaluronidase and partially to trypsin and elastase (Pasquali Ronchetti et al., 1986). The results seem to confirm that at least some of the alterations of connective tissues in PXE are due to abnormal PGs metabolism and to their tendency to form abnormal aggregates in the extracellular space.


The plasma membrane redox system: a candidate source of aging-related oxidative stress.
The plasma membrane redox system (PMRS) is an electron transport chain in the plasma membrane that transfers electrons from either intra- or extracellular donors to extracellular acceptors. Unlike the superoxide-generating NADPH oxidase of phagocytes and the homologous (but much less active) enzymes found in some other cells, the PMRS is still incompletely characterised at the molecular level. Much is known, however, concerning its function and affinity for both physiological and non-physiological substrates. A role for it in aging, the 'reductive hotspot hypothesis' (RHH), was proposed in 1998 as part of an explanation for the apparently indefinite survival in vivo of cells that have entirely lost mitochondrial respiratory capacity as a result of the accumulation of mitochondrial mutations. Stimulation of the PMRS might allow the cell to maintain redox homeostasis even while continuing to operate the Krebs cycle, which may be advantageous in many ways. However, the PMRS may, like the mitochondrial respiratory chain, be prone to generate superoxide when thus dysregulated - and in this case superoxide would be generated outside the cell, where antioxidant defences are more limited than inside the cell and where much highly oxidisable material is present. Cascades of peroxidation chain reactions initiated by this process may greatly amplify the oxidative stress on the organism that is caused by rare mitochondrially mutant cells. Since such cells increase in abundance with aging (though remaining rare), this is an economical hypothesis to explain the rise in oxidative stress seen in (and generally believed to contribute substantially to) mammalian aging. In an extension of previously published accounts of RHH, I propose here that the lysosomal toxicity of oxidised cholesterol derivatives (oxysterols) may contribute to the toxicity of mitochondrial mutations by affecting lysosomal function in many cell types in the same way as they have been proposed to do in arterial macrophages.


Mycobacterial manipulation of the host cell.
Phagosome biogenesis, the process by which macrophages neutralize ingested pathogens and initiate antigen presentation, has entered the field of cellular mycobacteriology research largely owing to the discovery 30 years ago that phagosomes harboring mycobacteria are refractory to fusion with lysosomes. In the past decade, the use of molecular genetics and biology in different model systems to study phagosome biogenesis have made significant advances in understanding subtle mechanisms by which mycobacteria inhibit the maturation of its phagosome. Thus, we are beginning to appreciate the extent to which these pathogens are able to interfere with innate immune responses and manipulate defense mechanisms to enhance their survival within the human host cell. Here, we summarize current knowledge about phagosome maturation arrest in infected macrophages and the subsequent attenuation of the macrophage-initiated adaptive anti-mycobacterial immune defenses.


Managing variability in the summary and comparison of gait data.
Variability in quantitative gait data arises from many potential sources, including natural temporal dynamics of neuromotor control, pathologies of the neurological or musculoskeletal systems, the effects of aging, as well as variations in the external environment, assistive devices, instrumentation or data collection methodologies. In light of this variability, unidimensional, cycle-based gait variables such as stride period should be viewed as random variables and prototypical single-cycle kinematic or kinetic curves ought to be considered as random functions of time. Within this framework, we exemplify some practical solutions to a number of commonly encountered analytical challenges in dealing with gait variability. On the topic of univariate gait variables, robust estimation is proposed as a means of coping with contaminated gait data, and the summary of non-normally distributed gait data is demonstrated by way of empirical examples. On the summary of gait curves, we discuss methods to manage undesirable phase variation and non-robust spread estimates. To overcome the limitations of conventional comparisons among curve landmarks or parameters, we propose as a viable alternative, the combination of curve registration, robust estimation, and formal statistical testing of curves as coherent units. On the basis of these discussions, we provide heuristic guidelines for the summary of gait variables and the comparison of gait curves.


Effect of chloroquine on human immunodeficiency virus (HIV) vertical transmission.
Over 2 million children globally are HIV positive. More than 90% are infected in utero from their mothers. Current pharmacological methods to reduce the rate of vertical transmission are too expensive for the developing world. Chloroquine, a cheap, widely available drug, has anti-HIV properties. We conducted a pilot study to determine if chloroquine can reduce HIV vertical transmission. 287 samples of stored, frozen cord blood from a cohort of Ugandan infants born to HIV positive mothers were analyzed for concentrations of chloroquine and its two major metabolites, monodesethylchloroquine and didesethylchloroquine. The HIV status of each infant was determined by ELISA with Western Blot confirmation at 15 and 18 months of age. 49% of samples had measurable chloroquine or metabolite. Of those with measurable drug, the higher concentrations of chloroquine and its metabolites were more frequently associated with HIV negative infants. However, only the median concentration of didesethylochloroquine was significantly higher in HIV negative infants vs. HIV positive infants (1.6 ng/ml vs. 0.9 ng/ml, p=0.05). Nearly half of all infants in a Ugandan cohort are exposed to chloroquine in the last trimester of pregnancy. Such random maternal chloroquine use may be associated with a decreased rate of HIV vertical transmission. The issue of maternal chloroquine use requires controlled study before any clinical conclusions may be drawn.


Birefringence measurement of rapid structural changes during collagen denaturation.
Linear birefringence, an optical property that results from a material's structure and composition, can be used to study dynamic changes in tissue structure. Single, 200 microseconds-long pulses from a Ho:YAG laser emitting 2.1 microns radiation were used to induce changes in the linear birefringence of rat tail tendon. Such changes were measured on a millisecond timescale. The measured rate coefficients describing the denaturation are not predicted by previous studies of collagen denaturation induced by slower, lower-temperature heating. Two types of laser-induced collagen denaturation can be differentiated: thermal denaturation, which appears rate-limited, and thermomechanical denaturation, which is observed at higher laser radiant exposures. Neither process is described by standard Arrhenius-type kinetic models.


Urethral Mobilization as an Alternative Procedure for Distal Hypospadias Repair.
To evaluate the urethral mobilization procedure as the main technique in management of glanular, coronal, subcoronal, and distal penile hypospadias. Sixty patients with distal hypospadias were included in this prospective study between January 2013 and January 2015. All of them had urethral mobilization. All patients were followed up at 2 weeks, and at 1, 3, 6, and 12 months postoperatively. Duration of the operation, hospital stay, position of meatus and its shape before and after the operation, and the postoperative complications were recorded. The force and direction of the postoperative voiding stream were observed and assessed subjectively. The mean operative time and hospital stay were 38 minutes and 2.2 days, respectively. Postoperatively, 2 patients developed wound infection, 1 of them developed glanular suture dehiscence managed by simple glanular suturing 3 months later. None of our patients developed urethrocutaneous fistula. Three months postoperatively, 2 patients developed meatal stenosis, 1 of them managed by meatal dilation whereas the other patient required ventral meatotomy. Hypospadias surgery aim to construct a functionally and cosmetically normal penis with minimal complications. Recently, urethral mobilization started to regain its position in distal hypospadias repair. Its results are comparable with other popular procedures or even better. Urethral mobilization is an easy and simple procedure and it has an excellent cosmetic and functional outcome with minimal complications.


Measure of total body water, using infrared spectrometry of heavy water.
The total body water can be assayed by the measurement of the diffusion space of heavy water. A well reproducible and inexpensive method is the infrared spectrometry, using two differential comparisons.


Investigating the relationship between physical activity and self-reported outcomes in adults with rotator cuff related shoulder pain.
Rotator cuff related shoulder pain is common among older adults. While exercise is often recommended for this condition, the relationship between physical activity levels and self-reported outcomes is unclear. This study investigated whether self-reported outcomes (pain, function, depressive symptoms, self-efficacy, general mental and physical health) relate to physical activity, and whether those who participated in shoulder-specific exercise had better self-reported outcomes than those who performed whole-body exercise. Forty-six participants with rotator cuff related shoulder pain participated from which 35 had complete datasets. Questionnaires were used to assess physical activity, pain, physical function, general health, and self-efficacy. Physical activity levels were also measured using an accelerometer. Neither pain nor other self-reported outcomes were related to subjective or objective physical activity levels. Participants that completed shoulder-specific exercise had significantly higher exercise self-efficacy than those who completed nonspecific exercise (P = .01; d = 0.91). A significant relationship between pain or self-reported outcomes and physical activity was not identified. Those who self-reported regularly exercising their injured shoulder had higher exercise self-efficacy than those who did not. These findings have clinical implications, suggesting that strategies to boost exercise self-efficacy may be important for older adults with rotator cuff related shoulder pain.


Red-winged blackbird predation on periodical cicadas (Cicadidae: Magicicada spp.): bird behavior and cicada responses.
Predation by red-winged blackbirds Agelaius phoeniceus L. on 13-year periodical cicadas (Magicicada spp.) and reactions by periodical cicadas to predators were studied during emergence of Brood XIX during summer of 1985 in northwestern Arkansas (USA). Emergences of periodical cicadas are classic examples of predator satiation due to high local densities of cicadas and birds are the major predators of adult periodical cicadas. Reactions of periodical cicadas to predators were assessed by recording behaviors exhibited by cicadas when approached in trees by a human hand during the 3-week period of peak adult densitics. Most male cicadas made a noise in association with escape behaviors when approached, and 50% of the females, which are silent, attempted escape behaviors. Observations of predation attempts on cicadas by red-winged blackbirds were made during the period of peak predation pressure. Red-winged blackbirds spent less time searching for cicadas over that 2-week period of increasing predation pressure, and became increasingly efficient at capturing cicadas. Handling time of cicadas by red-winged blackbirds increased by about 20 seconds over that period, as blackbirds spent more time consuming female cicadas. The flysquawk response, used only by male cicadas, was effective in deterring red-winged blackbirds; only 5% of the attacks by blackbirds were successful when that behavior was exhibited. All cicadas that remained motionless and silent when approached by blackbirds were captured and consumed. Because females remained inactive when approached more often than did males, blackbirds may have consumed more female cicadas. Changes that appeared in reactions of preiodical cicadas to the model predator and to the attacks of blackbirds reflect both changes in the sex ratio of the cicada populations and changes in behaviors of cicadas associated with mating and egg laying. The loud noise made by male periodical cicadas at mating centers did not appear to deter predation by blackbirds. Changes in the behavior of blackbirds that appeared to be in association with greater predation on female periodical cicadas relates directly to aspects of foraging theory, particularly predictions concerning more selective foraging during periods of abundant food resources.


Treatment Patterns, Depression, and Anxiety Among US Patients Diagnosed with Hyperhidrosis: A Retrospective Cohort Study.
Hyperhidrosis is associated with social and emotional stress due to limitations on health-related quality of life. This study examined real-world treatment patterns and concomitant depression and/or anxiety in patients with hyperhidrosis. Commercial health plan members in the US with ≥ 2 hyperhidrosis diagnosis codes and/or antiperspirant prescription claims were identified from January 2010 through November 2017. A control cohort (CC) of patients without hyperhidrosis was matched to the hyperhidrosis cohort on demographic characteristics. Depression and/or anxiety were identified by ≥ 1 relevant diagnosis code or pharmacy claim. A multivariable logistic regression model estimated odds of treatment in the hyperhidrosis cohort, and depression/anxiety in the hyperhidrosis cohort and CC, adjusting for patient characteristics. A total of 44,484 patients with hyperhidrosis were identified, of whom 58.5% were female, with a mean (± standard deviation) age of 36.5 ± 16.5 years (83.5% ≥ 18 years). A small majority of patients (51.6%, 0.69/person-year) received treatment with prescription antiperspirants. Post-index oral systemic therapies, medical procedures, and surgical options were uncommon. At 12 months post-index, 48.4% of the sample had not filled a prescription for extra- or prescription-strength antiperspirants. Compared with the CC (n = 137,451), a higher percentage of patients with hyperhidrosis had depression or anxiety reported during follow-up (41.1 vs. 28.2%, p < 0.001); this corresponded to higher adjusted odds of depression/anxiety in patients with hyperhidrosis [odds ratio (OR) 1.76, 95% confidence interval (CI) 1.72-1.80, p < 0.001]. Baseline depression and/or anxiety were associated with lower odds of receiving hyperhidrosis treatment (OR 0.77, 95% CI 0.73-0.80), as was increasing age and male gender. Patients with hyperhidrosis also had more frequent incident depression/anxiety during follow-up (18.2 vs. 10.6%, p < 0.001). In this real-world analysis, hyperhidrosis was associated with increased odds of depression and/or anxiety. However, relatively low percentages of patients received prescription topical or oral treatments or underwent surgery, suggesting that tolerability, efficacy, and provider awareness may be limiting factors in the effective treatment of hyperhidrosis.


A simple spectrophotometric method for estimation of plasma angiotensin I converting enzyme activity.
The procedure described is rapid, fairly simple, and inexpensive. The method may easily be used even in a small clinical laboratory. This method, based on a specific reaction which gives a product, hippuric acid azlactone, which absorbs in the visible region, avoids the interference of reagents and solvents encountered in the UV spectrophotometric assay, in which hippuric acid is determined directly.


The adverse effect of chocolate on lower esophageal sphincter pressure.
Decreased lower esophageal sphincter (LES) pressure after ingestion of chocolate has been previously noted. We have further evaluated the effect of chocolate on the known ability of gastric alkalinization or bethanechol to increase LES tone. 9 normal subjects were studied using an infused open-tip recording system. Pressure was monitored for a 15-min basal period, and for 60 min after ingestion of 120 ml of chocolate syrup either alone or with the concurrent administration of commercial antacid, oral bethanechol, or subcutaneous bethanechol. After chocolate ingestion, mean basal LES pressure of 14.6 +/- 1.1 (+/-SEM) mm Hg decreased significantly (P less than 0.01) to 7.9 +/- 1.3 mm Hg. An identical LES response occurred when antacid was given with the chocolate dose. Oral bethanechol (25 mg) and chocolate together resulted in lesser decreases in LES pressure. Subcutaneous bethanechol (5 mg) and chocolate produced significant increases (P less than 0.05) in sphincter pressure, although of lesser magnitude than reported with bethanechol alone. These results indicate that the adverse effect of chocolate on the LES is not reversed by gastric alkalinization and suggest that bethanechol in sufficient dose may overcome chocolate-induced decreases in LES pressure.


Characterization of the human peripheral lung adenosine receptor.
Because adenosine narrows asthmatic airways, is released during hypoxia and by mast cells, and is antagonized by theophylline, it may play a role in asthma. I characterized the first step in pulmonary responses to adenosine: its adenosine receptor. Plasma membranes, prepared from macroscopically normal human peripheral lung, were incubated with 10 nM 5'-N-ethylcarboxamido[3H]adenosine ([3H]NECA) and various concentrations of competing ligand under experimentally determined optimal conditions: 4 degrees C, pH 7.4, 5 mM MgCl2, 1.8 mg protein/ml, 30-min incubation time. Bound and free ligand were separated by rapid vacuum filtration, and the radioactive counts were analyzed using a weighted, non-linear, least-squares curve-fitting program, LIGAND. Analyzed together, the data from the lungs of 6 patients revealed a single binding site with a dissociation constant (Kd) for NECA of 200 nM +/- 14% and a receptor concentration of 543 fmol/mg protein +/- 13%. Analyzed separately, the individual Kds ranged from 133 to 430 nM and the receptor concentrations from 338 to 811 fmol/mg protein. Adenosine receptor ligands competed with NECA in an A2 rank order of potency: NECA greater than 8-phenyltheophylline greater than 3-isobutyl-1-methylxanthine greater than theophylline greater than N6-L-phenylisopropyladenosine greater than N6-D-phenylisopropyladenosine greater than N6-cyclohexyladenosine. Theophylline bound to the receptor with an inhibition constant (Ki = 70.9 microM +/- 28%) near the therapeutic range (28 to 56 microM). Cromolyn also bound with high affinity (Ki = 5.42 microM +/- 47%). I conclude that human lung adenosine receptors: (1) are single-site receptors, probably of the A2 subtype and (2) bind to both theophylline and cromolyn.


American Joint Committee on Cancer staging system 7th edition versus 8th edition: any improvement for patients with squamous cell carcinoma of the tongue?
In this study, we evaluated the 8th edition of American Joint Committee on Cancer (AJCC) staging criteria and lymph node ratio (LNR) to identify patients affected by squamous cell carcinoma of the tongue (SCCT) with a poor prognosis. Seventy-three cases of SCCT were analyzed retrospectively. Tumor staging was revised according to the 7th and 8th editions of the AJCC criteria. Depth of invasion (DOI), extranodal extension (ENE), and LNR were evaluated. Twenty-five cases were reclassified: 17 patients received an upstage in the staging score, and in 8 cases in the same stage group, pT or pN was changed. In the pT-upstaged group, 7 patients experienced recurrence, and 8 died. In the pN-upstaged group, 9 patients developed recurrence, and 10 died. The number of disease recurrence or death was higher in the groups of patients who received an upstage in pN and in the staging score (P < .05). The pN-upstaged group showed worse disease-free survival (DFS) and overall survival (OS) (P < .05). LNR was higher in patients with recurrence, and among these, LNR was lower in patients with ENE (P <.05). The 8th edition of the AJCC criteria allows for better stratification of patients with SCCT. The implementation of ENE and LNR to pN classification seems to identify patients with worse DFS and OS.


Direct electrochemistry of novel affinity-tag immobilized recombinant horse heart cytochrome c.
During the last decade protein electrochemistry at miniaturized electrodes has become important not only for functional studies of the charge transfer properties of redox proteins but also for fostering the development of sensitive biosensor and bioelectronic devices. One of the major challenges in this field is the directed coupling between electronic and biologically active components. A prerequisite for a fast and reversible electron transfer between electrode and protein is that the protein can be bound to the electrode in a favourable orientation. We examined electrostatic and bioaffinity-tag binding strategies for the directed immobilization of horse heart cytochrome c (cytc) on gold electrode surfaces to achieve this goal. Horse heart cytc was expressed in E. coli either as non-modified or genetically modified, i.e. histidine (his)-tag containing protein. The his-tags were introduced at defined positions at the N- or C-terminus of the polypeptide. It was our aim to generate tagged-versions of cytc that facilitate strong electronic coupling between protein and electrode and, at the same time, retain their catalytic and regulatory properties. The combination of different immobilization strategies, e.g. his-tag and electrostatic immobilization also opens new avenues for bivalent immobilization of proteins. This is of interest for molecular bioelectronic and biosensing applications where the proteins are immobilized between two crossing electrodes.


[Bile duct involvement in autoimmune pancreatitis: classification and treatment].
Autoimmune pancreatitis is a rare disease characterized by inflammation of the pancreatic parenchyma, irregular narrowing of the pancreatic duct, periductal lymphoplasmacytic infiltration and fibrosis at histological examination, the presence of autoantibodies and hypergammaglobulinemia, as well as the possible association of cholangitis and other autoimmune diseases. There is a favorable response to steroid therapy. We report the case of a patient with autoimmune pancreatitis with bile duct involvement and peripheral eosinophilia, requiring long-term immunosuppressant treatment. The diagnosis of a diffuse form of AIP was made without direct histological evidence and based on indirect imaging, clinical and laboratory findings in an autoimmune context. The histological and imaging studies of bile duct involvement and the favourable response to steroids were additional arguments.


Low serum vitamin D is associated with axial length and risk of myopia in young children.
The aim of the study was to investigate the relationship between serum 25(OH)D levels and axial length (AL) and myopia in 6-year-old children. A total of 2666 children aged 6 years participating in the birth-cohort study Generation R underwent a stepwise eye examination. First, presenting visual acuity (VA) and AL were performed. Second, automated cycloplegic refraction was measured if LogMAR VA > 0.1. Serum 25-hydroxyvitamin D [25(OH)D] was determined from blood using liquid chromatography/tandem mass spectrometry. Vitamin D related SNPs were determined with a SNP array; outdoor exposure was assessed by questionnaire. The relationships between 25(OH)D and AL or myopia were investigated using linear and logistic regression analysis. Average 25(OH)D concentration was 68.8 nmol/L (SD ± 27.5; range 4-211); average AL 22.35 mm (SD ± 0.7; range 19.2-25.3); and prevalence of myopia 2.3 % (n = 62). After adjustment for covariates, 25(OH)D concentration (per 25 nmol/L) was inversely associated with AL (β -0.043; P < 0.01), and after additional adjusting for time spent outdoors (β -0.038; P < 0.01). Associations were not different between European and non-European children (β -0.037 and β -0.039 respectively). Risk of myopia (per 25 nmol/L) was OR 0.65 (95 % CI 0.46-0.92). None of the 25(OH)D related SNPs showed an association with AL or myopia. Lower 25(OH)D concentration in serum was associated with longer AL and a higher risk of myopia in these young children. This effect appeared independent of outdoor exposure and may suggest a more direct role for 25(OH)D in myopia pathogenesis.


A model for heterogeneous nuclear ribonucleoproteins in telomere and telomerase regulation.
The heterogeneous nuclear ribonucleoproteins (hnRNPs) are a large family of nucleic acid binding proteins that are often found in, but not restricted to, the 40S-ribonucleoprotein particle. Subsets of hnRNPs are strictly nuclear while others shuttle between the nucleus and cytoplasm. Members of the hnRNP family have been implicated to have roles in many aspects of mRNA maturation/turnover and in telomere and telomerase regulation. Telomeres are repetitive DNA elements mainly found at the ends of human chromosomes. In most normal cells, telomeres shorten with each cell division. Telomere shortening can be compensated for by a ribonucleoprotein complex, called telomerase. Telomerase, consisting of an integral RNA and catalytic protein component as well as several auxiliary factors, extends the 3'-G-rich strand of the ends of the telomeres. Here we present new data and describe a model that implicates the telomerase bound hnRNPs in promoting telomere access by interacting with telomeres. Telomere bound hnRNPs include hnRNP A1, A2-B1, D and E and telomerase bound hnRNPs including hnRNPA1 C1/C2 and D. The telomere and telomerase bound hnRNPs may prove to be good targets for regulating telomere length.


Annual dynamic dataset of global cropping intensity from 2001 to 2019.
The cropping intensity has received growing concern in the agriculture field in applications such as harvest area research. Notwithstanding the significant amount of existing literature on local cropping intensities, research considering global datasets appears to be limited in spatial resolution and precision. In this paper, we present an annual dynamic global cropping intensity dataset covering the period from 2001 to 2019 at a 250-m resolution with an average overall accuracy of 89%, exceeding the accuracy of the current annual dynamic global cropping intensity data at a 500-m resolution. We used the enhanced vegetation index (EVI) of MOD13Q1 as the database via a sixth-order polynomial function to calculate the cropping intensity. The global cropping intensity dataset was packaged in the GeoTIFF file type, with the quality control band in the same format. The dataset fills the vacancy of medium-resolution, global-scale annual cropping intensity data and provides an improved map for further global yield estimations and food security analyses.


Glycogen-rich and glycogen-depleted astrocytes in the oedematous human cerebral cortex associated with brain trauma, tumours and congenital malformations: an electron microscopy study.
The anaerobic mobilization of astrocyte glycogen in anoxic-ischemic regions of the oedematous human cerebral cortex is analysed. Seventeen cortical biopsies of patients with brain trauma, brain tumours and congenital malformations were examined by conventional transmission electron microscopy. Glycogen-rich and glycogen-depleted, clear or dense astrocytes cell bodies were observed in anoxic ischaemic regions of different brain cortical areas in perineuronal, neuropilar and perivascular localization. Glycogen-rich astrocytes showed clear or moderately dense cytoplasm and accumulation of both beta-type or monogranular glycogen granules and alpha-type or multigranular glycogen particles. Focal regions of translucent cytoplasm were observed in areas of glycogen degradation. Glycogen-depleted astrocytes exhibited a clear cytoplasm and scarce amount or absence of beta-type glycogen granules. Coexisting glycogen-rich and glycogen-depleted neuropilar astrocytic processes were observed in the vicinity of degenerated myelinated axons and degenerated axodendritic contacts. Glycogen-rich and glycogen-depleted perivascular astrocytic processes were also found surrounding injured and collapsed cerebral capillaries. The findings suggest astrocytic glycogen mobilization during anoxic and ischaemic conditions, revealing the important contribution of astrocytes on neuronal survival under conditions of energy substrate limitations.


Ethylene response factor ERF022 is involved in regulating Arabidopsis root growth.
Ethylene response factors (ERFs) are involved in the regulation of plant development processes and stress responses. In this study, we provide evidence for the role of ERF022, a member of the ERF transcription factor group III, in regulating Arabidopsis root growth. We found that ERF022-loss-of-function mutants exhibited increased primary root length and lateral root numbers, and also morphological growth advantages compared to wild-type. Further studies showed that mutants had enhanced cell size in length in the root elongation zones. These results were accompanied by significant increase in the expression of cell elongation and cell wall expansion related genes SAUR10, GASA14, LRX2, XTH19 in mutants. Moreover, ERF022-mediated root growth was associated with the enhanced endogenous auxin and gibberellins levels. Our results suggest that loss-of-function of ERF022 up-regulated the expression of cell elongation and cell wall related genes through auxin and gibberellins signal in the regulation of root growth. Unexpectedly, ERF022 overexpression lines also showed longer primary roots and more lateral roots compared to wild-type, and had longer root apical meristematic zone with increased cell numbers. Overexpression of ERF022 significantly up-regulated cell proliferation, organ growth and auxin biosynthesis genes EXO, HB2, GALK2, LBD26, YUC5, which contribute to enhanced root growth. Altogether, our results provide genetic evidence that ERF022 plays an important role in regulating root growth in Arabidopsis thaliana.


Hypoxia down-regulates secretion of MMP-2, MMP-9 in porcine pulmonary artery endothelial and smooth muscle cells and the role of HIF-1.
Primary cell culture, techniques of gene transfection, gelatin zymography, and Western blot were used to investigate the effect of hypoxia on the secretion of MMP-2 and MMP-9 in pulmonary artery endothelial cells (PAEC) and smooth muscle cells (PASMC), and the role of HIF-1. Our results showed that (1) after exposure to hypoxia for 24 h, the protein content and activity of MMP-2 in the PAEC medium as well as these of MMP-2 and MMP-9 in PASMC medium (P < 0. 01) decreased significantly in contrast to those in normoxic group (P < 0.05); (2) after transfection of wild type EPO3'-enhancer, a HIF-1 decoy, the content and activity of MMP-2 and MMP-9 in hypoxic mediums became higher than those in normoxic group (P < 0.01), while transfection of mutant EPO3'-enhancer didn't affect the hypoxia-induced down-regulation. It is concluded that hypoxia could inhibit the secretion and activity of MMP-2 and MMP-9 in PAEC and PASMC, which could be mitigated by the transfection of EPO3'-enhancer and that HIF-1 pathway might contribute to hypoxia-induced down-regulation of MMP-2 and MMP-9.


5-Hydroxymethyl-2-furaldehyde induces developmental toxicology and decreases bone mineralization in zebrafish larvae.
In this study, we aimed to assess the developmental toxicity and effects of 5-HMF in zebrafish as a model organism for toxicology studies. To this end, we treated zebrafish embryos with 1-100 μg/ml 5-HMF and observed bone staining, gene expression, and reactive oxygen species levels in order to investigate the toxicological effects of 5-HMF. The results showed that high concentrations of 5-HMF caused increased mortality and deformity rates in zebrafish larvae, inhibited cartilage development, reduced bone mineralization, increased reactive oxygen species levels, and disrupted the expression of genes related to bone development and reactive oxygen species enzyme activity. The antioxidant N-acetyl-l-cysteine partially rescued the toxicological effects caused by the high concentrations of 5-HMF. Overall, these findings showed that high concentrations of 5-HMF induce reactive oxygen species production, leading to developmental toxicity and decreased bone mineralization. Our results provide a reference for understanding the toxic effects of 5-HMF.


Landscape heterogeneity increases the stability of wild ungulate populations facing climatic variability in Mediterranean ecosystems.
Mediterranean environments are characterized by strong intra- and inter-annual fluctuations in plant primary production, which are likely to regulate the carrying capacity and density-dependent responses of ungulate populations. These effects may, however, vary across spatial and temporal scales. Habitat heterogeneity, particularly when associated to differentiated phenological responses, may allow wild ungulates to mitigate temporal fluctuations in plant production by using different resources along the year. In this work, we use a 15-years dataset (including remote-sensing data on vegetation distribution, phenology and production, as well as ungulate population counts) to assess how temporal variability in plant primary production and livestock abundance influence the population dynamics of two wild ungulates: native red deer, Cervus elaphus, and introduced fallow deer, Dama dama. Results show that temporal alternation in the phenological cycles of the four different vegetation types increased plant production, thus food availability for ungulates, within each year. Furthermore, complementarity in the responses of different vegetation types to variations in the amount and timing of rainfall increased the predictability of food availability across different years. This complementarity effect was further increased by the contrasting responses of ungulate populations to variation in the production of different vegetation types. Furthermore, domestic ungulates had positive effects on wild ungulate density at low to intermediate abundances, but high livestock densities decreased ungulate density and constrained the stability of the plant-ungulate system in response to the impact of climatic variation, particularly under climate change. Our findings deepen the knowledge on vegetation-ungulate interactions in Mediterranean areas, potentially contributing to develop better management strategies of ungulate populations and adapt them to ongoing climate change.


Studies on the genetic basis of human influenza A virus adaptation to mice: degrees of virulence of reassortants with defined genetic content.
A highly virulent mouse-adapted variant of influenza virus A/Aichi/2/68 (H3N2) was crossed either with the original A/USSR/90/77 (H1N1) influenza virus strain or with its mouse-adapted, moderately mouse virulent variant. The reassortants were characterized with respect to their genetic content and pneumovirulence for mice. The reassortants fell into three categories: avirulent, highly virulent (resembling in this respect the parent A/Aichi/2/68 virus) and moderately virulent (resembling the mouse-adapted A/USSR/90/77 parent virus). The analysis of the parental origin of the genes of 6 reassortants allowed to suggest that changes in the HA gene and in a polymerase gene (most likely, PB1) were necessary for the acquisition of virulence by the A/USSR/90/77 virus in the course of adaptation to mice, whereas the changes in two other polymerase genes as well as in the genes NA and NS were not involved. The low degree of pathogenicity characteristic of the mouse-adapted A/USSR/90/77 virus was determined by gene(s) other than HA.


Metabolic network model guided engineering ethylmalonyl-CoA pathway to improve ascomycin production in Streptomyces hygroscopicus var. ascomyceticus.
Ascomycin is a 23-membered polyketide macrolide with high immunosuppressant and antifungal activity. As the lower production in bio-fermentation, global metabolic analysis is required to further explore its biosynthetic network and determine the key limiting steps for rationally engineering. To achieve this goal, an engineering approach guided by a metabolic network model was implemented to better understand ascomycin biosynthesis and improve its production. The metabolic conservation of Streptomyces species was first investigated by comparing the metabolic enzymes of Streptomyces coelicolor A3(2) with those of 31 Streptomyces strains, the results showed that more than 72% of the examined proteins had high sequence similarity with counterparts in every surveyed strain. And it was found that metabolic reactions are more highly conserved than the enzymes themselves because of its lower diversity of metabolic functions than that of genes. The main source of the observed metabolic differences was from the diversity of secondary metabolism. According to the high conservation of primary metabolic reactions in Streptomyces species, the metabolic network model of Streptomyces hygroscopicus var. ascomyceticus was constructed based on the latest reported metabolic model of S. coelicolor A3(2) and validated experimentally. By coupling with flux balance analysis and using minimization of metabolic adjustment algorithm, potential targets for ascomycin overproduction were predicted. Since several of the preferred targets were highly associated with ethylmalonyl-CoA biosynthesis, two target genes hcd (encoding 3-hydroxybutyryl-CoA dehydrogenase) and ccr (encoding crotonyl-CoA carboxylase/reductase) were selected for overexpression in S. hygroscopicus var. ascomyceticus FS35. Both the mutants HA-Hcd and HA-Ccr showed higher ascomycin titer, which was consistent with the model predictions. Furthermore, the combined effects of the two genes were evaluated and the strain HA-Hcd-Ccr with hcd and ccr overexpression exhibited the highest ascomycin production (up to 438.95 mg/L), 1.43-folds improvement than that of the parent strain FS35 (305.56 mg/L). The successful constructing and experimental validation of the metabolic model of S. hygroscopicus var. ascomyceticus showed that the general metabolic network model of Streptomyces species could be used to analyze the intracellular metabolism and predict the potential key limiting steps for target metabolites overproduction. The corresponding overexpression strains of the two identified genes (hcd and ccr) using the constructed model all displayed higher ascomycin titer. The strategy for yield improvement developed here could also be extended to the improvement of other secondary metabolites in Streptomyces species.


A method to quantify several tyrosine kinase inhibitors in plasma by micellar liquid chromatography and validation according to the European Medicines Agency guidelines.
A procedure based on micellar liquid chromatography has been developed to monitor five tyrosine kinase inhibitors in plasma, prescribed against several kinds of cancer: erlotinib, imatinib, sunitinib, sorafenib and lapatinib. The sample was diluted in a micellar solution and directly injected, thus clean-up steps were not required. The analytes were resolved without interferences in <20 min using a C18 column and a mobile phase of 0.13 M SDS-4% 1-butanol, buffered at pH 3.5, running under isocratic mode at 1 mL/min. The detection was performed by UV-visible absorbance, using a wavelength program to maximize the signal-to-noise ratio. The method was validated following the guideline of the European Medicines Agency in terms of: selectivity, calibration range (0.05-5 μg/mol), linearity (r(2)>0.990), limit of detection (15-35 ng/mL), carry-over effect, accuracy (-10.4 to +11.0%), precision (<9.2%), matrix effect, robustness (<8.4%) and stability. The procedure is rapid, easy-to-handle, uses a low amount of toxic chemical provide reliable results. Finally, the method was successfully used to analyze the studied tyrosine kinase inhibitors in plasma from cancer patients.


Uncovering Philip Morris International's Fundamental Strategies for Product Placement in Thailand: Spotlighting Industry Penetration to Advance the Endgame.
Tobacco companies and their associated businesses know that placement - where one can see and purchase their products - is critical to their success. Placement is one of the four fundamental Ps of marketing along with product, price and promotion. Placement includes identifying retail locations in important places such as in shopping districts, within neighborhoods, near schools, at beaches, and in parks. In Southeast Asia, counteracting tobacco company placement strategies that result in market penetration is essential to advancing the endgame, namely ending tobacco use. However, in Southeast Asia research on the placement of tobacco products has been limited. We undertook to analyze how Philip Morris International (PMI) through its subsidiary Philip Morris Asia Inc. (PMAI), from the time the company entered Thailand's market once it was forced open in 1990, developed its successful product placement strategies and tactics. We examined over 4,000 PMI and PMAI internal documents using an historical, iterative and thematic approach. We analyzed the most relevant and illuminating documents, particularly those in which PMAI discussed retailer supply, retailer acceptance and retailer cooperation. Even before foreign tobacco brands were legally allowed to be sold in Thailand, PMAI was already doing customer research in Thailand. In 1989, PMAI conducted a study of potential Thai customers in which 24% of respondents' lack of availability (i.e., product placement) was one of the main reasons for not smoking PMI's products. Based on these findings, PMAI engaged in intensive internal efforts to address the placement barrier to gain share. PMAI placed considerable emphasis on "stimulating retail trade acceptance" by making payments to retailers who met agreed upon and contracted product sales targets. PMAI's initial successes incentivizing Thai retailers by essentially buying prime retail space for placement of their brands, to crowd out local and other foreign brands, became the foundation of what evolved into a sophisticated program to make placement highly lucrative for retailers. PMAI viewed aggressive product placement as essential to success as a new entrant in Thailand, and their product placement strategies contributed substantially to capturing a large share of the market. Therefore, endgame strategies must focus on restricting product placement through surveillance of tobacco industry legal, investment and retailer actions and through stricter tobacco retailer licensing requirements and penalties.


Scavenging of photogenerated ROS by Oxicams. Possible biological and environmental implications.
The profusely employed drugs Piroxicam (Piro), Tenoxicam (Teno) and Meloxicam (Melo) belonging to the non-steroidal antiinflammatory drug (NSAID) family of the Oxicams (Oxis) were studied in the frame of two specific conditions: (a) their ROS scavenging ability, in relation to a possible biological antioxidant action and (b) their photodegradability under environmental conditions, in the context of Oxi-contaminated waters. Singlet molecular oxygen (O2((1)Δg)) and superoxide radical anion (O2(-)) were photogenerated through Riboflavin (Rf, vitamin B2)-photosensitization in aqueous and aqueous-methanolic solutions in the presence of Oxi concentrations in the range 50-500 μM. The visible-light absorber vitamin is currently present in all types of natural waters and constitutes the most frequent endogenous photosensitizer in mammals. Hence, it was employed in order to mimic both natural sceneries of interest. All three Oxis quench O2((1)Δg) with rate constants in the order of 10(8)M(-1)s(-1) showing a significant photodegradation efficiency given by a dominant reactive fashion for deactivation of the oxidative species. Although this is not a desirable property in the context of photoprotection upon prolonged photoirradiation, constitutes in fact a promissory aspect for the degradation NSAIDs, in waste waters. Indirect evidence indicates that Melo is also oxidized through a O2(-)-mediated component. The simultaneous presence of Piro plus tryptophan or tyrosine under Rf-photosensitizing conditions, which has taken the amino acids as photooxidizable model residues in a proteinaceous environment, indicates that the NSAID induces a protection of the biomolecules against photodynamic degradation.


Enhanced cell killing, inhibition of recovery from potentially lethal damage and increased mutation frequency by 3-aminobenzamide in Chinese hamster V79 cells exposed to X-rays.
X-ray induced potentially lethal damage and its inhibition by the aromatic amide 3-aminobenzamide have been investigated in Chinese hamster V79 cells. 3-Aminobenzamide (3-AB) is a known inhibitor of polyadenosine diphosphoribose synthetase. With increasing concentrations of 3-AB an increasing inhibition of PLD repair was observed. Little inhibition of PLD repair was seen when 3-AB was added 3 h following irradiation. Utilizing the 6-thioguanine mutation assay, the effect of poly(ADP-R) synthetase inhibition under conditions of PLD repair upon mutation frequency were also studied. A large increase in mutation frequency following 24 h post-irradiation recovery in the presence of 3-AB was seen. These results favour a possible role of 3-AB in preventing repair by facilitating early damage fixation before repair can occur, simultaneously reducing G2-arrest.


Identification of bacteriophage phi 29 prohead RNA domains necessary for in vitro DNA-gp3 packaging.
Functional domains of the bacteriophage phi 29 prohead RNA (pRNA) that are essential for in vitro packaging of DNA-gp3 into the prohead were mapped using pRNA mutants. Oligonucleotide-directed mutant pRNAs were produced that contained deletions and sequence alterations but were predicted to retain the overall secondary structure of wild-type pRNA. Mutant pRNAs were compared to wild-type pRNA for prohead binding in a competition assay and for DNA packaging in the defined in vitro system. The prohead binding site was previously localized to residues 22-84 on the 120-residue domain I of pRNA by ribonuclease footprinting (Reid, R. J. D., Bodley, J. W., and Anderson, D. (1994) J. Biol. Chem. 269, 5157-5162). Mutations of pRNA within the prohead binding site resulted in substantial loss of prohead binding capacity, while mutations outside of the footprint had moderate effects on prohead binding. DNA-gp3 packaging activity was correlated with pRNA binding activity for mutations within the footprint. This mutational analysis showed that both sequence and secondary structure of residues 40-91 of pRNA were crucial for prohead binding and that elements of the A helix formed from residues 1-28 and 117-92 were needed for DNA packaging functions other than prohead binding.


Usefulness and safety of cimetidine in patients receiving mexiletine for ventricular arrhythmia.
Cimetidine, a commonly used H2 receptor antagonist, was found to adversely interact with many drugs metabolized by the liver, including class I antiarrhythmic agents, lidocaine and quinidine. Mexiletine is a new class I antiarrhythmic agent similar to lidocaine which when used orally may have significant gastric side effects. Since some patients with peptic ulcer disease or gastric hyperacidity on mexiletine may benefit from the addition of cimetidine, it was important to rule out any significant adverse interaction between the two drugs in such patients. Eleven patients currently receiving long-term oral mexiletine for the treatment of complex ventricular arrhythmia underwent a double-blind crossover trial where they were maintained on their usual dose of mexiletine, and cimetidine, 300 mg orally every 6 hours, or placebo were added for a 1-week period each. Peak and trough mexiletine blood levels were not significantly altered by cimetidine. Similarly, there was no significant change in the frequency and severity of ventricular arrhythmia when cimetidine was added to mexiletine. Cimetidine reduced gastric side effects of mexiletine in 50% of patients who had complained of such symptoms on mexiletine alone or on mexiletine and placebo. We conclude that cimetidine can effectively reduce gastric side effects of mexiletine in many patients without adversely affecting the plasma concentration or the efficacy of the drug.


The applicability of correction factor for corneal thickness on non-contact tonometer measured intraocular pressure in LASIK treated eyes.
To determine the applicability of central corneal thickness (CCT) based correction factor for non-contact tonometer (NCT) measured intraocular pressure (IOP) readings. A prospective, non-randomized study involved 346 eyes of 173 consecutive patients with age ⩾21 years undergoing laser-assisted in situ keratomileusis (LASIK) for myopia and/or myopic astigmatism. The CCT and IOP were measured before and after the LASIK procedure. The IOP pre and post-LASIK was compared after applying the correction factor for CCT. Patients not completing the 3 month postoperative follow-up were excluded. The median spherical equivalent before undergoing LASIK was -4.25D (inter-quartile range, -3.25D). The mean preoperative CCT was 536.82 ± 33.71 μm which reduced to 477.55 ± 39.3 μm (p < 0.01) post-LASIK. The mean IOP reduced from a preoperative value of 14.6 ± 2.32 mmHg to 10.64 ± 2.45 mmHg postoperatively (p < 0.01). On applying correction for the corneal thickness, the pre and postoperative IOP was 15.14 ± 2.8 mmHg and 15.37 ± 2.65 mmHg (p = 0.06) respectively with a strong positive correlation (r = 0.7, p < 0.01). Three hundred eyes (86.7%) had an absolute difference in IOP of less than 3.0 mmHg post-CCT correction which is within the retest variability of NCT. Only 46 eyes (13.3%) had an absolute difference of more than 3.0 mmHg. The modified Ehler's correction algorithm used in this study can be effectively applied in the normal IOP range in a majority of patients.


Factors Influencing Selection of Active Surveillance for Localized Prostate Cancer.
To determine how well demographic and clinical factors predict the initiation of Active Surveillance (AS). AS has been suggested as a way to reduce overtreatment of men who have prostate cancer; however, factors associated with the decision to choose AS are poorly quantified. Using the Michigan Urological Surgery Improvement Collaborative registry, we identified 2977 men with prostate cancer who made treatment decisions from January 1, 2012, through December 31, 2013. We used chi-square and Wilcoxon tests to examine the association between factors and initiation of AS. Logistic regression models were fit for D'Amico risk categories. Measures of model discrimination and calibration were estimated, including area under the curve (AUC) and Brier score (BS). Patient age, Gleason score, clinical T-stage, urology practice, and tumor volume (greatest percent of a core involved with cancer and proportion of positive cores) were associated with the decision to choose AS in the intermediate-risk cohort (AUC = 0.875, BS = 0.07) and the complete cohort (AUC = 0.89, BS = 0.10). Patient age, urology practice, and tumor volume were significant in the low-risk cohort (AUC = 0.71, BS = 0.22). The addition of urology practice increased AUC in the low-risk cohort from 0.71 to 0.76 and reduced BS from 0.22 to 0.21. The urology practice at which a patient is seen is an important predictor for whether patients will initiate AS. Predictions were least accurate for low-risk patients, suggesting that factors such as patient preference play a role in treatment decisions.


Switchable reactivity of 2-benzoyl glycals towards stereoselective access of 1-3 and 1-1 S/O linked disaccharides.
We have developed a synthesis of 1-3 and 1-1 disaccharides from 2-benzoyl glycal and anomeric thiol and/or hydroxy sugar acceptors under mild conditions at room temperature. The regio and stereo-selectivity of the newly formed inter-glycosidic linkages are dependent on the nature of the glycal donor (D or L) and anomeric acceptor.


Ketotifen-induced nocturnal bruxism.
Nocturnal bruxism is a common oromandibular movement disorder highly prevalent in children, but its pathophysiological mechanism has not been fully explained. Iatrogenic sleep bruxism has been described following treatment with several psychotropic medications. However, no case of antihistamine-induced bruxism has been reported to date. Herein, we describe a 4-year-old child who experienced nocturnal bruxism during treatment for bronchospasm and rhinitis with the antihistamine ketotifen. Drug rechallenge was also performed. The present case adds useful information to our knowledge of bruxism. Complex and poorly understood interactions between multiple central nervous system neurotransmitters, such as histamine, serotonin, and dopamine, are involved.


Post-transplant Lymphoproliferative Disorder in Recipient's Reconstructed Middle Hepatic Vein After Pediatric Living Donor Liver Transplant: A Case Report.
Post-transplant lymphoproliferative disorder (PTLD) is known as one of the most frequent post-transplant neoplastic diseases, which may lead to recipient and graft morbidity after liver transplant. A 14-year-old boy received pediatric living donor liver transplant (right living graft) 29 months ago, with etiology of biliary atresia. During 22-month follow-up after transplant, computed tomography and positron emission tomography with computed tomography scan showed a single progressive mass locating in the graft cutting surface with occlusive reconstructed middle hepatic vein (MHV). Serology was negative for Epstein-Barr virus serology. Mild fever and pneumonia did not improve after 1-week intravenous antibiotics. PTLD was considered. A surgical resection was scheduled and performed in compliance with the Helsinki Congress and the Istanbul Declaration. During laparotomy, a single mass located in the reconstructed MHV from segment V to the inferior vena cava was confirmed. Postoperative immunohistochemical result showed CD 3(+), CD 20(+), CD 38(+), CD 10(-), CD 56(-), Ki-67(+, 20%-30%), Epstein-Barr virus-encoded RNA(-), and Epstein-Barr virus nuclear antigen 2(-). Polymorphic PTLD was eventually diagnosed. No recurrence or new set lesions were detected after 6-month follow-up. This is the first case describing PTLD may originate from reconstructed MHV after pediatric living donor liver transplant. As a life-threatening complication of liver transplant, surgical resection should be considered as a safe and feasible treatment for the single resectable mass.


Prophylaxis of atopic disease by six months' total solid food elimination. Evaluation of 135 exclusively breast-fed infants of atopic families.
One hundred and thirty-five infants of atopic parents were exclusively breast-fed for 6 months without any cow's milk based supplements. Of these infants 70 received no nourishment except breast milk during the 6 months, and 65 were started on solid foods at the age of 3 months. The diet of all the infants was similar during 6 to 12 months of age. The children were examined at the age of one year. In the exclusive breast milk group atopic eczema and food allergy were less frequent than in the solid food group. The results suggest that total solid food elimination for the first 6 months of life, in addition to exclusive breast milk feeding, is prophylactic for atopic disease in children who are at hereditary risk.


Suppressive effect of ultraviolet (UVB and PUVA) radiation on superantigen production by Staphylococcus aureus.
It is well known that Staphylococcus aureus (S. aureus) proliferates on the moist skin lesion of atopic dermatitis. Reduction of bacteria colonization from skin lesions by antibiotics has been reported to be effective for the treatment of atopic dermatitis. S. aureus produces superantigens which can activate T cells and possibly enhance the inflammatory reaction. Photo(chemo)therapy has been successfully used for the treatment of severe cases of atopic dermatitis. We have previously reported that photo(chemo)therapy had bacteriostatic effect on S. aureus. Now we examined the effect of UVB and psoralen plus UVA (PUVA) on superantigen production from S. aureus. We isolated S. aureus from six atopic dermatitis patients. S. aureus was irradiated in vitro with UVB (0, 5, 10 mJ/cm2) or PUVA (0.001% psoralen plus 0, 5, 10 mJ/cm2 UVA) and incubated 4 h with 100 strokes per min. After incubation, the amounts of superantigens in the supernatant were measured using ELISA kit. The production of superantigens decreased in an ultraviolet dose-dependent manner. The suppressive effects of UV radiation on superantigen production may be involved in the therapeutic efficacy of photo(chemo)therapy for atopic dermatitis.


Identification and validation of a novel anoikis-related signature for predicting prognosis and immune landscape in ovarian serous cystadenocarcinoma.
Ovarian serous cystadenocarcinoma (OSC) is the most prevalent histological subtype of ovarian cancer (OV) and presents a serious threat to women's health. Anoikis is an essential component of metastasis, and tumor cells can get beyond it to become viable. The impact of anoikis on OSC, however, has only been the topic of a few studies. The mRNA sequencing and clinical information of OSC came from The Cancer Genome Atlas Target Genotype-Tissue Expression (TCGA TARGET GTEx) dataset. Anoikis-related genes (ARGs) were collected by Harmonizome and GeneCards websites. Centered on these ARGs, we used unsupervised consensus clustering to explore potential tumor typing and filtered hub ARGs to create a model of predictive signature for OSC patients. Furthermore, we presented clinical specialists with a novel nomogram based on ARGs, revealing the underlying clinical relevance of this signature. Finally, we explored the immune microenvironment among various risk groups. We identified 24 ARGs associated with the prognosis of OSC and classified OSC patients into three subtypes, and the subtype with the best prognosis was more enriched in immune-related pathways. Seven ARGs (ARHGEF7, NOTCH4, CASP2, SKP2, PAK4, LCK, CCDC80) were chosen to establish a risk model and a nomogram that can provide practical clinical decision support. Risk scores were found to be an independent and significant prognostic factor in OSC patients. The CIBERSORTx result revealed an inflammatory microenvironment is different for risk groups, and the proportion of immune infiltrates of Macrophages M1 is negatively correlated with risk score (rs = -0.21, P < 0.05). Ultimately, quantitative reverse transcription polymerase chain reaction (RT-PCR) was utilized to validate the expression of the seven pivotal ARGs. In this study, based on seven ARGs, a risk model and nomogram established can be used for risk stratification and prediction of survival outcomes in patients with OSC, providing a reliable reference for individualized therapy of OSC patients.


The retro-inverso form of a homeobox-derived short peptide is rapidly internalised by cultured neurones: a new basis for an efficient intracellular delivery system.
The capacity of a retro-inverso form of a 16-residue peptide from the Antennapedia homeodomain to be taken up by cultured neurones was tested. Like its homologue made of L-amino acids, it was rapidly internalised and distributed throughout the cytoplasm and even the cell nucleus. The amount of retro-inverso peptide in cells after incubation in culture medium was 3.4 times that of the L-peptide form. With a cholesteryl moiety attached to the C-terminus to increase its lipophilicity, the retro-inverso peptide was internalised 8 times better than the L-form. The greater efficiency of the peptidomimetic is probably due to the resistance to proteolytic degradation conferred by the D-amino acids, as the sequence contains two sites sensitive to neuronal endoproteases. The peptide might be the basis for development of system for delivering a variety of molecules into cells.


Peripheral T-cell lymphoma in childhood. A clinicopathological study of six cases.
A review of the pathological material from 42 children with non-Hodgkin's lymphoma seen over a 44 month period revealed 10 large cell tumours. Of these, six were classified as peripheral T-cell lymphoma, an entity rarely reported in childhood. Three patients were boys and three girls (median age 9.5 years), and extranodal presentation was a feature of two patients. Five had high-grade tumours; of these, three were classified as large cell anaplastic, Ki-1 positive and two as pleomorphic large cell. The remaining patient had a low-grade tumour of angioimmunoblastic type. T-cell subsets were examined in three cases and showed the following phenotypes: CD4-, CD8-; CD4+, CD8-; CD4-, CD8+. Three of the patients with high-grade tumours died, with a mean survival of 22 weeks. The remaining patients are alive and clinically disease-free for between 10 and 24 months after treatment.


Revisiting the basis for US ballast water regulations.
The transport and release of invasive organisms in ballast water has harmed ecosystems, economic activities and human health. Current US ballast water regulations intended to minimize the discharge of such organisms are based on results reported by a scientific advisory committee in 2011. Using the same methods, we re-analyzed the data evaluated by the committee as well as new data. We find that the committee's analysis was flawed, and that some treatment systems can meet limits that are 10 times (for zooplankton) or 1000 times (for phytoplankton) more stringent than the committee reported. These findings suggest that US ballast water standards, and similar standards in a recently ratified international agreement, should be re-evaluated.


Cortical maps: where theory meets experiments.
Primary visual cortex (V1) has remarkably systematic functional maps. One commonly used class of computational models proposes that such maps are generated by a mechanism that projects the multiple dimensions of neuronal responses smoothly onto the two dimensions of cortex. In this issue of Neuron, Mriganka Sur and colleagues find a close match between such model predictions and measurements from ferret V1.


Implementation of Kangaroo Care in a Pediatric Cardiac Intensive Care Unit.
With supporting the best neurodevelopmental outcomes possible, the doctor of nursing practice project was constructed. An educational program was created for nursing staff discussing the importance of kangaroo care and how to safely facilitate it in the intensive care setting. Following the education completion, kangaroo care was implemented. Data were collected exploring barriers to implementation, discharge on maternal breast milk, and parental feedback.


Kras Gene Analysis Using Liquid-Based Cytology Specimens Predicts Therapeutic Responses and Prognosis in Patients with Pancreatic Cancer.
Although several molecular analyses have shown that the Kras gene status is related to long-term survival of patients with pancreatic ductal adenocarcinoma (PDAC), the results remain controversial. Here, we examined the Kras gene status in a cohort of unresectable PDAC patients who underwent first-line therapy with gemcitabine and nab-paclitaxel (GA) and assessed differences in chemotherapy responses and survival. Patients with a histological diagnosis of PDAC (based on EUS-guided fine-needle aspiration) from 2017 to 2019 were enrolled. Tumor genomic DNA was extracted from residual liquid-based cytology specimens and Kras mutations were assessed using the quenching probe method. The relationships between the Kras status and progression-free survival (PFS) and overall survival (OS) were assessed. Of the 110 patients analyzed, 15 had wild-type Kras. Those with the wild-type gene showed significantly longer PFS and OS than those with mutant Kras (6.9/5.3 months (p = 0.044) vs. 19.9/11.8 months (p = 0.037), respectively). Multivariate analyses identified wild-type Kras as a significant independent factor associated with longer PFS and OS (HR = 0.53 (p = 0.045) and HR = 0.35 (p = 0.007), respectively). The analysis of the Kras gene status could be used to predict therapeutic responses to GA and prognosis in unresectable PDAC patients.


The Sforzesco brace and SPoRT concept: a brace to replace cast in worst curves.
Bracing still remains the most important conservative treatment for scoliosis; approaches to bracing are really many and diverse. The aim of this paper of this paper, is the presentation of the brace we recently developed (the Sforzesco brace and SPoRT concept) to face worst scoliosis, in the past treated through casting. The SPoRT concept was born while we were looking for a new brace; it means Symmetric, Patient oriented, Rigid, Three-dimensional and Active. Details of construction and the biomechanical principle of its corrective action are reported. This brace has been used widely in the last 3 years. The Sforzesco brace showed to be more effective than the Lyon brace and as effective as the Risser cast on different clinical and radiological outcome measures in 2 different prospective study of best clinical practice. When compared to the other braces, the Sforzesco can find its place in the most important curves, in which there are no alternatives to try avoiding surgery. The SPoRT Concept appear a useful tool to interpret what happens with this brace, even if in the future could be overcome by new theories according to new (and awaited) results in the literature.


Prediction of treatment response in patients with newly diagnosed type 2 diabetes: the Skaraborg diabetes register.
Type 2 diabetes is associated with cardiovascular complications. It is largely unknown which patients have poor treatment response and high complication risk; biomarkers are studied for this purpose. The aim of the study was to investigate the association between clinical factors such as HbA1c, level of biomarkers (C-peptide, copeptin) at diagnosis and changes in HbA1c, blood pressure or body mass index (BMI) after five years. Clinical data and blood samples from 460 newly diagnosed type 2 diabetes patients from the Skaraborg diabetes register (SDR) at diagnosis and after 5years and were analyzed with linear and logistic regressions. High BMI at diagnosis and smoking were associated with less reduction of HbA1c i.e. poorer treatment outcome after 5years. A high HbA1c at baseline predicted a greater reduction of HbA1c and need for insulin treatment. High systolic blood pressure and BMI at baseline were associated with greater reduction. The biomarkers were not associated with increase of blood pressure, HbA1c, BMI or need for insulin treatment. Smokers and patients with high HbA1c at diagnosis respond poorer to treatment over 5years. This highlights the importance of advice for non-smoking and weight reduction and more intensive treatment over time.


Sleep in Children With Neurodevelopmental Disabilities During COVID-19: An Integrative Review.
Sleep issues occur at higher rates in children with neurodevelopmental disorders than in the typical population. Little is known about the impact of COVID-19 on sleep issues in this population METHOD: This integrative review aimed to characterize studies during the COVID-19 pandemic (2020-2022) addressing the prevalence and management of sleep issues in children and youth with neurodevelopmental disorders.Comprehensive database searches were used to identify articles, and 31 studies were considered suitable for this review. Most studies reported worsening sleep disturbances during COVID-19 restrictions. All studies were conducted when clinics were closed; only two studies addressed treatment options such as music therapy and sleep hygiene education and found improvement in sleep issues. Future research needs to concentrate on developing interventions to assist families remotely and empower families with a toolkit of preparedness in times of crisis.