Microbiota associations with inflammatory pathways in asthma.
The airway microbiota plays an important role in asthma pathophysiology. However, the relationship between the airway microbiota and asthma phenotypes is still poorly understood. We aimed to characterize the airway microbiota in asthma patients and determine its correlation with airway inflammatory phenotypes and other phenotypic characteristics. The microbial composition of induced sputum specimens collected from asthma patients was determined using 16S rDNA gene sequencing. Patients with asthma had a higher abundance of bacterial taxa associated with Bacteroidetes, Fusobacteria and Proteobacteria and a reduced abundance of Firmicutes and Actinobacteria compared to healthy controls. This study classified the asthma-associated lung microbiota into three community types based on DMM models, which were defined as three pulmotypes (P1, P2 and P3). The lungs of patients with pulmotype 3 (P3) were dominated by Faecalibacterium and Bacteroides, while patients with pulmotype 1 (P1) had a greater abundance of Pasteurellaceae, Streptococcus and Rothia. P1 patients were older (p = .045) and had lower blood TGF levels (p = .028). P3 patients had fewer eosinophils (p = .016) and more neutrophils (p = .039) in induced sputa than P1 patients. Differences in asthma-associated airway microbiota pulmotypes are associated with and might influence asthma, particularly inflammatory phenotypes.


Co-localization of P450 enzymes in the rat substantia nigra with tyrosine hydroxylase.
Susceptibility to develop Parkinson's disease has been linked to abnormalities of P450 enzyme function. Multiple P450 enzymes are expressed in brain but the relationship of these to Parkinson's disease is unknown. We have investigated the expression of P450 enzymes in the rat substantia nigra and their co-localization in tyrosine hydroxylase-positive neurons and astrocytes. Immunohistochemistry was performed using anti-peptide antisera against the following P450 enzymes: CYP1A1, CYP1A2, CYP2B1/2, CYP2C12, CYP2C13/2C6, CYP2D1, CYP2D4, CYP2E1, CYP3A1, CYP3A2 and NADPH-P450 oxidoreductase. Immunoreactivity in nigral cells was found only for CYP2E1 and CYP2C13/2C6. CYP2E1 immunoreactivity was localized to many midbrain nuclei including the substantia nigra pars compacta but not the substantia nigra pars reticulata while immunoreactivity to CYP2C13/2C6 was found in the substantia nigra pars compacta, substantia nigra pars reticulata and many other midbrain nuclei. Sections of rat midbrain double labelled for either CYP2E1 or CYP2C13/2C6 and tyrosine hydroxylase or glial fibrillary acidic protein were examined for co-localization by confocal laser scanning microscopy. CYP2E1 and CYP2C13/2C6 immunoreactivity was found in tyrosine hydroxylase-positive neurons in the substantia nigra pars compacta but not in glial cells. CYP2C13/2C6, but not CYP2E1, was also found in non-glial, non-tyrosine hydroxylase-expressing cells in the substantia nigra pars reticulata. Isoniazid induction increased CYP2E1 fluorescence signal intensity from nigral dopaminergic neurons. At least two P450 enzymes are found in nigral dopamine containing cells and one, namely CYP2E1, is selectively localized to this cell population. CYP2E1 is a potent generator of free radicals which may contribute to nigral pathology in Parkinson's disease. The expression of CYP2E1 in dopaminergic neurons in substantia nigra raises the possibility of a causal association with Parkinson's disease.


Effect of modified genioglosuss advancement on hyoid bone position: Cephalometric study.
To assess Hyoid bone position and retrolingual airway space after Modified Genioglossus Advancement Surgery by cephalometry in patients with obstructive sleep apnea (OSA). Prospective study. Zagazig University Hospital. Eighteen patients with moderate to severe OSA having multilevel airway obstruction confirmed by fiberoptic endoscopy during Muller's maneuver and DISE. All patients underwent modified genioglossus advancement surgery associated with antrolateral advancement pharyngoplasty. Beside Polysomnography and Drug induced sleep endoscopy, Cephalometry was done preoperatively and 6 months postoperative. Improved Polysomnography parameters as Postoperative mean ± SD apnea hypopnea index decreased from 52 ± 17.1 to 17 ± 3 (P < 0.001, 95% confidence interval 27.71 to 42.41). LOS increased from 79.89 ± 4.43% to 83 ± 4.05% (P 0.07, 95% confidence interval -0.31 to 6.97). Cephalometry analysis showed a significant difference between preoperative and postoperative findings, including: Retrolingual airway space at three levels significantly increased; Level 1 from 6.1 ± 1.6 to 8.5 ± 1.7, Level 2 from 10.5 ± 2.4 to 13.9 ± 2.1, Level 3 from 15.7 ± 3.1 to 21 ± 4, H-GN decreased from 51 ± 7 to 39 ± 8, H-MP decreased from 31.6 ± 7.7 to 24.9 ± 7.3, HS decreased from 121 ± 15 to 102 ± 12, H-PH increased from 29 ± 8 to 43 ± 9. With a success rate defined as AHI <20 and a 50% decrease in AHI of the preoperative value, the surgical success rate was 83.33%. This study showed that Modified genioglossus advancement procedures done for OSA patients significantly changed the position of hyoid bone into a more anterior and superior position and this was reflected in the postoperative Polysomnography.


High osmolarity and L-DOPA augment release via the fusion pore in PC12 cells.
We have amperometrically measured dopamine release from rat pheochromocytoma cells (PC12 cells) in high osmolarity conditions with and without L-3,4-dihydroxyphenylalanine (L-DOPA) treatment. We observe an increase in the number of release events displaying a prespike feature or "foot" when the cells are stimulated in high osmolarity saline. We also see an increase in foot area and duration when cells are stimulated in high osmolarity saline, or high osmolarity saline subsequent to incubation with the dopamine precursor L-DOPA in isotonic saline, which serves to increase the vesicle size. The data suggest that membrane biophysics are an important component in defining the rate, duration and amount of neurotransmitter release via the fusion pore.


Widespread frontoparietal fMRI activity is greatly affected by changes in criterion placement, not discriminability, during recognition memory and visual detection tests.
Widespread frontoparietal activity is consistently observed in recognition memory tests that compare studied ("target") versus unstudied ("nontarget") responses. However, there are conflicting accounts that ascribe various aspects of frontoparietal activity to mnemonic evidence versus decisional processes. According to Signal Detection Theory, recognition judgments require individuals to decide whether the memory strength of an item exceeds an evidence threshold-the decision criterion-for reporting previously studied items. Yet, most fMRI studies fail to manipulate both memory strength and decision criteria, making it difficult to appropriately identify frontoparietal activity associated with each process. In the current experiment, we manipulated both discriminability and decision criteria across recognition memory and visual detection tests during fMRI scanning to assess how frontoparietal activity is affected by each manipulation. Our findings revealed that maintaining a conservative versus liberal decision criterion drastically affects frontoparietal activity in target versus nontarget response contrasts for both recognition memory and visual detection tests. However, manipulations of discriminability showed virtually no differences in frontoparietal activity in target versus nontarget response or item contrasts. Comparing across task domains, we observed similar modulations of frontoparietal activity across criterion conditions, though the recognition memory task revealed larger activations in both magnitude and spatial extent in these contrasts. Nonetheless, there appears to be some domain specificity in frontoparietal activity associated with the maintenance of a conservative versus liberal criterion. We propose that widespread frontoparietal activity observed in target versus nontarget contrasts is largely attributable to response bias where increased activity may reflect inhibition of a prepotent response, which differs depending on whether a person maintains a conservative versus liberal decision criterion.


Effects of dietary amino acids and repeated handling on stress response and brain monoaminergic neurotransmitters in Senegalese sole (Solea senegalensis) juveniles.
The present study aimed to assess the effects of increased availability of dietary amino acids (AA) on brain monoamine neurotransmitters and the metabolic processes resulting from stressful situations in fish. Senegalese sole (Solea senegalensis) juveniles (24.2±0.4g wet mass) were weekly subjected to an acute handling stressor (HDLG) or remained undisturbed (CTL). Additionally, both treatments were fed a control or a high protein (HP) diet (CTL, CTL HP, HDLG and HDLG HP). The HP diet slightly increased the levels of digestible indispensable AA, together with tyrosine and cysteine. Repeated handling induced a stress response after 14 and 28 days in fish held at both HDLG and HDLG HP treatments. While dietary treatment and handling stress activated the serotonergic system at 14 days, these effects were not observed after 28 days. In addition, the HP diet minimized the decrease in plasma indispensable AA due to repeated handling stress after 28 days. It was concluded that HP diet decreased post-stress plasma glucose and lactate levels in HDLG HP specimens only at 14 days of treatment. Moreover, dietary treatment was also effective in stimulating DA synthesis and release, thus dietary phenylalanine supplementation can increase DA biosynthesis in fish.


The Chinese highly pathogenic porcine reproductive and respiratory syndrome virus infection suppresses Th17 cells response in vivo.
Porcine reproductive and respiratory syndrome virus (PRRSV) has been shown to immunomodulate innate and adaptive immunity of pigs. The Chinese highly pathogenic PRRSV (HP-PRRSV) infection causes severe bacterial secondary infection in pigs. However, the mechanism in relation to the bacterial secondary infection induced by HP-PRRSV remains unknown. In the present study, Th17 cells response in peripheral blood, lungs, spleens and lymph nodes of piglets were analyzed, and bacterial loads in lungs of piglets were examined upon HP-PRRSV infection. Meanwhile the changes of CD4(+) and CD8(+) T cells in peripheral blood of the inoculated piglets were analyzed. The results showed that HP-PRRSV-inoculated piglets exhibited a suppressed Th17 cells response in peripheral blood and a reduced number of Th17 cells in lungs, and higher bacterial loads in lungs, compared with low pathogenic PRRSV. Moreover, HP-PRRSV obviously resulted in severe depletion of porcine T cells in peripheral blood at the early stage of infection. These findings indicate that HP-PRRSV infection suppresses the response of Th17 cells that play an important role in combating bacterial infections, suggesting a possible correlation between the suppression of Th17 cells response in vivo and bacterial secondary infection induced by HP-PRRSV. Our present study adds a novel insight into better understanding of the pathogenesis of the Chinese HP-PRRSV.


TALAIA: a 3D visual dictionary for protein structures.
Graphical analysis of the molecular structure of proteins can be very complex. Full-atom representations retain most geometric information but are generally crowded, and key structural patterns can be challenging to identify. Non-full-atom representations could be more instructive on physicochemical aspects but be insufficiently detailed regarding shapes (e.g. entity beans-like models in coarse grain approaches) or simple properties of amino acids (e.g. representation of superficial electrostatic properties). In this work, we present TALAIA a visual dictionary that aims to provide another layer of structural representations.TALAIA offers a visual grammar that combines simple representations of amino acids while retaining their general geometry and physicochemical properties. It uses unique objects, with differentiated shapes and colors to represent amino acids. It makes easier to spot crucial molecular information, including patches of amino acids or key interactions between side chains. Most conventions used in TALAIA are standard in chemistry and biochemistry, so experimentalists and modelers can rapidly grasp the meaning of any TALAIA depiction. We propose TALAIA as a tool that renders protein structures and encodes structure and physicochemical aspects as a simple visual grammar. The approach is fast, highly informative, and intuitive, allowing the identification of possible interactions, hydrophobic patches, and other characteristic structural features at first glance. The first implementation of TALAIA can be found at https://github.com/insilichem/talaia.


On the relevance of technical variation due to building pools in microarray experiments.
Pooled samples are frequently used in experiments measuring gene expression. In this method, RNA from different individuals sharing the same experimental conditions and explanatory variables is blended and their concentrations are jointly measured. As a matter of principle, individuals are represented in equal shares in each pool. However, some degree of disproportionality may arise from the limits of technical precision. As a consequence a special kind of technical error occurs, which can be modelled by a respective variance component. Previously published theory - allowing for variable pool sizes - has been applied to four microarray gene expression data sets from different species in order to assess the practical relevance of this type of technical error in terms of significance and size of this variance component. The number of transcripts with a significant variance component due to imperfect blending was found to be 4329 (23 %) in mouse data and 7093 (49 %) in honey bees, but only 6 in rats and none whatsoever in human data. These results correspond to a false discovery rate of 5 % in each data set. The number of transcripts found to be differentially expressed between treatments was always higher when the blending error variance was neglected. Simulations clearly indicated overly-optimistic (anti-conservative) test results in terms of false discovery rates whenever this source of variability was not represented in the model. Imperfect equality of shares when blending RNA from different individuals into joint pools of variable size is a source of technical variation with relevance for experimental design, practice at the laboratory bench and data analysis. Its potentially adverse effects, incorrect identification of differentially expressed transcripts and overly-optimistic significance tests, can be fully avoided, however, by the sound application of recently established theory and models for data analysis.


Predictive model for reduction of Escherichia coli during acetic acid decontamination of chicken skin.
The response surface methodology was used to evaluate the effect of operating variables (acetic acid concentration, spraying time and temperature) on the reduction of Escherichia coli populations on poultry breast skin in a laboratory showering process, as well as to identify the best conditions that are required to develop this operation. Skin samples were inoculated with a 24-h E. coli culture and afterwards treated according to experimental design under selected acetic acid concentration, spraying time, and solution temperature. The E. coli reduction model was significantly affected by the acetic acid concentration and spraying time (P < or = 0.05 and < or =0.01), while temperature did not show a significant effect (P > 0.05). The predictive model obtained was validated through additional confirmatory experiments and showed to be adequate, and it could be used as an approach to optimize the acetic acid spray washes during poultry carcasses processing. The use of acetic acid washes in the processing of poultry does not have the capability of eliminating E. coli populations from carcasses. However, significant reductions in the initial load could be achieved.


Functional divergence of the sarcomeric myosin, MYH7b, supports species-specific biological roles.
Myosin heavy chain 7b (MYH7b) is an evolutionarily ancient member of the sarcomeric myosin family, which typically supports striated muscle function. However, in mammals, alternative splicing prevents MYH7b protein production in cardiac and most skeletal muscles and limits expression to a subset of specialized muscles and certain nonmuscle environments. In contrast, MYH7b protein is abundant in python cardiac and skeletal muscles. Although the MYH7b expression pattern diverges in mammals versus reptiles, MYH7b shares high sequence identity across species. So, it remains unclear how mammalian MYH7b function may differ from that of other sarcomeric myosins and whether human and python MYH7b motor functions diverge as their expression patterns suggest. Thus, we generated recombinant human and python MYH7b protein and measured their motor properties to investigate any species-specific differences in activity. Our results reveal that despite having similar working strokes, the MYH7b isoforms have slower actin-activated ATPase cycles and actin sliding velocities than human cardiac β-MyHC. Furthermore, python MYH7b is tuned to have slower motor activity than human MYH7b because of slower kinetics of the chemomechanical cycle. We found that the MYH7b isoforms adopt a higher proportion of myosin heads in the ultraslow, super-relaxed state compared with human cardiac β-MyHC. These findings are supported by molecular dynamics simulations that predict MYH7b preferentially occupies myosin active site conformations similar to those observed in the structurally inactive state. Together, these results suggest that MYH7b is specialized for slow and energy-conserving motor activity and that differential tuning of MYH7b orthologs contributes to species-specific biological roles.


Cloning and expression analysis of the chicken DEAD box gene DDX1.
DEAD box proteins are putative RNA unwinding proteins found in organisms ranging from mammals to bacteria. While some DEAD box genes expressed in higher eukaryotes are ubiquitous, others have distribution profiles that suggest a cell-, tissue-, or developmental-specific role. The DEAD box gene, DDX1, was identified by differential screening of a subtracted retinoblastoma cDNA library. A limited survey of human fetal tissues indicated that DDX1 mRNA has a widespread distribution but is not uniformly expressed in all tissues. To further document the spatial and temporal distribution of DDX1 during embryonic development, we cloned the chicken DDX1 cDNA. The predicted amino acid sequence of chicken DDX1 was 93% identical to that of human DDX1. All DEAD box motifs, as well as a SPRY domain, were present in chicken DDX1. Northern and Western blot analyses showed highest levels of DDX1 at early stages of development. Tissue maturation was generally accompanied by a decrease in expression, although DDX1 levels remained elevated in late embryonic retina and brain. In situ hybridization of retinal tissue sections revealed widespread distribution of DDX1 mRNA at early developmental stages with preferential expression in amacrine and ganglion cells of the differentiated tissue. Preferential expression of DDX1 was also observed in specific areas of the brain in older embryos, such as the external granule layer of the cerebellum. These results suggest a specific role for DDX1 in subsets of differentiated cells as well as a more general role in undifferentiated cells.


Safety and feasibility study of ex vivo expanded allogeneic-NK cells infusion in patients with acute pneumonia caused by COVID-19.
NK cells are the most active innate immune cells in antiviral immunity, which are impaired by SARS-COV2 infection. Infusion of allogeneic NK cells might be a complementary treatment to boost immune system function in COVID-19 patients. In this project, we focused on COVID-19 patients with low inspiratory capacity (LIC). This project aims to evaluate the feasibility and safety of allogeneic NK cell infusion as an intervention for respiratory viral disease. A non-blind two arms pilot study was designed and conducted after signing the consent form. Ten matched patients, in terms of vital signs and clinical features, were enrolled in the control and intervention groups. Approximately 2 × 10^6 cells/kg of NK cells were prepared under GCP (good clinical practice) conditions for each patient in the intervention group. The control group was under the same conditions and drug regimen except for the treatment with the prepared cells. Then, infused intravenously during 20 min in the ICU ward of Masih Daneshvari Hospital. The clinical signs, serological parameters, and CTCAE (Common Terminology Criteria for Adverse Events) were recorded for safety evaluation and the feasibility of project management were evaluated via designed checklist based on CONSORT. There were no symptoms of anaphylaxis, hypersensitivity, significant changes in blood pressure, cardiovascular complications, and fever from injection time up to 48 h after cell infusion. The mean hospitalization period in the control and intervention groups was 10 and 8 days, respectively. The blood O2 saturation level was raised after cell infusion, and a significantly lower mean level of inflammatory enzymes was observed in the intervention group following discharge compared to the control group (p < 0.05). The inflammatory parameters differences at the discharge date in cell therapy group were highly negative. Intravenous infusion of ex vivo-expanded allogeneic NK cells was safe and feasible. However, the efficacy of this approach to reducing the severity of disease in COVID-19 patients with LIC could not be determined. Name of the registry: NKCTC. IRCT20200621047859N2. December 29, 2020. URL of trial registry record: https://www.irct.ir/trial/49382.


Alkali action on the urinary crystallization of calcium salts: contrasting responses to sodium citrate and potassium citrate.
Alkali therapy is used commonly to prevent recurrent stone formation in patients with distal renal tubular acidosis. We compared the effects of potassium citrate to those of sodium citrate in 6 well defined cases of incomplete distal renal tubular acidosis. The patients were studied during a control phase, during potassium citrate treatment (80 mEq. per day) and during sodium citrate treatment (80 mEq. per day) chosen in random order. Potassium citrate caused a decrease in urinary calcium and a significant increase in urinary citrate that resulted in a significant decrease in the urinary saturation of calcium oxalate. It did not alter the saturation of brushite and sodium urate. However, while sodium citrate also was able to increase the urinary citrate level, there was no decrease in the urinary calcium (owing to the increased sodium load). Thus, the urinary saturation of calcium oxalate did not decrease as much as with potassium citrate and the saturation of brushite increased significantly. Moreover, the urinary saturation of sodium urate increased significantly owing to the enhanced sodium excretion. The results suggest that potassium citrate therapy may retard the crystallization of calcium oxalate and may not cause calcium phosphate crystallization. In contrast, sodium citrate may have no effect or it sometimes may accentuate the crystallization of calcium salts. Thus, our study supports the potential clinical advantage of potassium citrate therapy over sodium alkali treatment in patients with incomplete distal renal tubular acidosis and recurrent calcium nephrolithiasis.


Effects of Nefiracetam, a novel pyrrolidone-type nootropic agent, on the amygdala-kindled seizures in rats.
Nefiracetam (NEF) is a novel pyrrolidonetype nootropic agent, and it has been reported to possess various pharmacologic effects as well as cognition-enhancing effects. The present study focused on the effects of NEF in amygdala-kindled seizures and its potential for antiepileptic therapy. Effects of NEF on fully amygdala-kindled seizures and development of amygdala-kindled seizures were investigated in rats and compared with those of levetiracetam (LEV), a pyrrolidone-type antiepileptic drug (AED). In fully amygdala-kindled rats, NEF (25, 50, and 100 mg/kg, p.o.) decreased afterdischarge induction, afterdischarge duration, seizure stage, and motor seizure duration in a dose-dependent manner. LEV (25, 50, and 100 mg/kg, p.o.) had no effects on afterdischarge induction and slightly decreased afterdischarge duration, whereas it markedly decreased seizure stage and motor seizure duration. In contrast to the results in fully amygdala-kindled rats, NEF (25 and 50 mg/kg/day, p.o.) had few or no effects on the development of amygdala-kindled seizures. As well as fully amygdala-kindled seizures, LEV (50 mg/kg/day, p.o.) markedly inhibited the development of behavioral seizures without reducing daily afterdischarge duration. Although NEF possesses potent anticonvulsant effects on fully amygdala-kindled seizures, it has few or no effects on the development of amygdala-kindled seizures. LEV shows marked anticonvulsant effects on both phases of kindling. In fully amygdala-kindled rats, NEF inhibits both electroencephalographic and behavioral seizures, whereas LEV inhibits only behavioral seizures. This double dissociation suggests that NEF has a distinct anticonvulsant spectrum and mechanisms from those of LEV.


Analysis of gene expression profile of TPM3-ALK positive anaplastic large cell lymphoma reveals overlapping and unique patterns with that of NPM-ALK positive anaplastic large cell lymphoma.
Anaplastic large cell lymphoma (ALCL) comprises a group of non-Hodgkin lymphomas characterized by the expression of the CD30/Ki-1 antigen. A subset of ALCL is characterized by chromosomal translocations involving the anaplastic lymphoma kinase (ALK) gene on chromosome 2. While the most common translocation is the t(2;5)(p23;q35) involving the nucleophosmin (NPM) gene on chromosome 5, up to 12 other translocations partners of the ALK gene have been identified. One of these is the t(1;2)(q25;p23) which results in the formation of the chimeric protein TPM3-ALK. While several of the signaling pathways induced by NPM-ALK have been elucidated, those involved in ALCLs harboring TPM3-ALK are largely unknown. In order to investigate the expression profiles of ALCLs carrying the NPM-ALK and TPM3-ALK fusions, we carried out cDNA microarray analysis of two ALCL tissue samples, one expressing the NPM-ALK fusion protein and the other the TPM3-ALK fusion protein. RNA was extracted from snap-frozen tissues, labeled with fluorescent dyes and analyzed using cDNAs microarray containing approximately 9,200 genes and expressed sequence tags (ESTs). Quantitative fluorescence RT-PCR was performed to validate the cDNA microarray data on nine selected gene targets. Our results show a significant overlap of genes deregulated in the NPM-ALK and TPM-ALK positive lymphomas. These deregulated genes are involved in diverse cellular functions, such as cell cycle regulation, apoptosis, proliferation, and adhesion. Interestingly, a subset of the genes was distinct in their expression pattern in the two types of lymphomas. More importantly, many genes that were not previously associated with ALK positive lymphomas were identified. Our results demonstrate the overlapping and unique transcriptional patterns associated with the NPM-ALK and TPM3-ALK fusions in ALCL.


Influence of the introduction of a triphenylphosphine group on the anticancer activity of a copper complex.
Aiming at obtaining new copper complexes with good cytotoxicity against cancer cells, triphenylphosphine (TPP) was introduced to obtain insight into the influence of the co-ligands. In this paper, two copper complexes, Cu(2-pbmq)(CH3OH)Br2 (1) and [Cu(2-pbmq)(TPP)Br]2 (2) were designed, synthesized, and characterized by X-ray crystallography, 2-((2-(pyrazin-2-yl)-1H-benzo[d]imidazol-1-yl)methyl))quinolone (2-pbmq), to investigate the influence of the TPP group on the anticancer activity of the metal complex. Although the presence of the TPP group diminished the intensity of the interaction properties of the complex with DNA, the in vitro anticancer activity and cellular uptake of the TPP-containing complex were markedly superior to those of its TPP-lacking counterpart. Detailed studies on the more potently cytotoxic complex 2 revealed that it accumulated in nucleus, arrested the cell cycle at the G0-G1 phase, causing mitochondrial dysfunction, involving the potential simultaneous mitochondrial membrane collapse, cellular ATP level depletion, and Ca2+ leakage, eventually inducing cell apoptosis. In summary, the introduction of a TPP group enhances the biological activity and cytotoxicity of the complex.


Age-related morphology trends of cortical sulci.
The age-related trends of the width and the depth of major cortical sulci were studied in normal adults. Ninety healthy subjects (47 males, 43 females) age 20-82 years were evaluated. Measurements of average sulcal width and depth in 14 prominent sulcal structures per hemisphere were performed with high-resolution anatomical MRI. The average sulcal width increased at a rate of about 0.7 mm/decade, while the average sulcal depth decreased at a rate of about 0.4 mm/decade. Sulcal age-related trends were found to be highly influenced by gender in the superior temporal, collateral, and cingulate sulci (P < 0.05), with males showing more pronounced age-related change in sulcal width than females. Sulcal structures located in multimodal cortical areas showed more profound age-related changes than sulcal structures in unimodal cortical areas (P < 0.05).


Partial reinforcement in human biofeedback learning.
This paper reviews the evidence for the efficacy of partial reinforcement in producing resistance to extinction in human biofeedback experiments. The methodological criteria necessary to demonstrate such effects are discussed, as is the status of the analogy of reinforcement and information feedback. It is suggested that the problem of maintaining responding in the absence of feedback should be tackled empirically rather than assuming the validity of findings from other areas of learning theory.


Prospective evaluation of kidney and liver disease in autosomal recessive polycystic kidney disease-congenital hepatic fibrosis.
We have previously published the characteristics of kidney and liver disease in a cohort of 73 individuals with molecularly confirmed autosomal recessive polycystic kidney disease-congenital hepatic fibrosis, based upon cross-sectional data. Here, we present prospective data on the same cohort. Comprehensive biochemical and imaging data on progression of kidney and liver disease in 60 of the 73 patients were prospectively collected at the NIH Clinical Center on multiple visits between 2003 and 2019. Of the 73 patients, 23 received a renal allograft at an average age of 17.5 years and 10 underwent liver transplantation at an average age of 20.3 years. Patients who presented perinatally and those who had corticomedullary disease required kidney transplantation significantly earlier. The mean eGFR slope in patients with corticomedullary disease was -1.6 ml/min/1.73 m2/y, in comparison to -0.6 ml/min/1.73 m2/y in those with medullary disease. Kidney size remained the same over time and normalized to the upper limit of normal by 20-25 years of age. The extent of renal disease on ultrasound remained largely unchanged; no patient progressed from the "medullary" to the "corticomedullary" group. There was no correlation between eGFR slope and kidney size. The synthetic function of the liver remained largely intact even in patients with advanced portal hypertension. Based on spleen length/height ratio, two thirds of patients had portal hypertension which remained stable in 39% and worsened in 61%. Patients with portal hypertension had lower platelet counts and relatively higher levels of AST, GGT, direct bilirubin and ammonia. The progression rates of kidney and liver disease were independent of each other. Patients with bi-allelic non-truncating PKHD1 variants had similar progression of kidney and liver disease in comparison to those who were compound heterozygous for a non-truncating and a truncating variant.


Automatic three-dimensional nasal and pharyngeal airway subregions identification via Vision Transformer.
Upper airway assessment requires a fully-automated segmentation system for complete or sub-regional identification. This study aimed to develop a novel Deep Learning (DL) model for accurate segmentation of the upper airway and achieve entire and subregional identification. Fifty cone-beam computed tomography (CBCT) scans, including 24,502 slices, were labelled as the ground truth by one orthodontist and two otorhinolaryngologists. A novel model, a lightweight multitask network based on the Swin Transformer and U-Net, was built for automatic segmentation of the entire upper airway and subregions. Segmentation performance was evaluated using Precision, Recall, Dice similarity coefficient (DSC) and Intersection over union (IoU). The clinical implications of the precision errors were quantitatively analysed, and comparisons between the AI model and Dolphin software were conducted. Our model achieved good performance with a precision of 85.88-94.25%, recall of 93.74-98.44%, DSC of 90.95-96.29%, IoU of 83.68-92.85% in the overall and subregions of three-dimensional (3D) upper airway, and a precision of 91.22-97.51%, recall of 90.70-97.62%, DSC of 90.92-97.55%, and IoU of 83.41-95.29% in the subregions of two-dimensional (2D) crosssections. Discrepancies in volume and area caused by precision errors did not affect clinical outcomes. Both our AI model and the Dolphin software provided clinically acceptable consistency for pharyngeal airway assessments. The novel DL model not only achieved segmentation of the entire upper airway, including the nasal cavity and subregion identification, but also performed exceptionally well, making it well suited for 3D upper airway assessment from the nasal cavity to the hypopharynx, especially for intricate structures. This system provides insights into the aetiology, risk, severity, treatment effect, and prognosis of dentoskeletal deformities and obstructive sleep apnea. It achieves rapid assessment of the entire upper airway and its subregions, making airway management-an integral part of orthodontic treatment, orthognathic surgery, and ENT surgery-easier.


Short-term outcomes in robot-assisted compared to laparoscopic colon cancer resections: a systematic review and meta-analysis.
Robot-assisted surgery is increasingly adopted in colorectal surgery. However, evidence for the implementation of robot-assisted surgery for colon cancer is sparse. This study aims to evaluate the short-term outcomes of robot-assisted colon surgery (RCS) for cancer compared to laparoscopic colon surgery (LCS). Embase, MEDLINE, and Cochrane Library were searched between January 1, 2005 and October 2, 2020. Randomized clinical trials and observational studies were included. Non-original literature was excluded. Primary endpoints were anastomotic leakage rate, conversion to open surgery, operative time, and length of hospital stay. Secondary endpoints were surgical efficacy and postoperative morbidity. We evaluated risk of bias using RoB2 and ROBINS-I quality assessment tools. We performed a pooled analysis of primary and secondary endpoints. Heterogeneity was assessed by I2, and possible causes were explored by sensitivity- and meta-regression analyses. Publication bias was evaluated by Funnel plots and Eggers linear regression test. The level of evidence was assessed by GRADE. Twenty studies enrolling 13,799 patients (RCS 1740 (12.6%) and LCS 12,059 (87.4%) were included in the meta-analysis that demonstrated RCS was superior regarding: anastomotic leakage (odds ratio (OR) = 0.54, 95% CI [0.32, 0.94]), conversion (OR = 0.31, 95% CI [0.23, 0.41]), overall complication rate (OR = 0.85, 95% CI [0.73, 1.00]) and time to regular diet (MD = - 0.29, 95% CI [- 0.56, 0.02]). LCS proved to have a shortened operative time compared to RCS (MD = 42.99, 95% CI [28.37, 57.60]). Level of evidence was very low according to GRADE. RCS showed advantages in colonic cancer surgery regarding surgical efficacy and morbidity compared to LCS despite a predominant inclusion of non-RCT with serious risk of bias assessment and a very low level of evidence.


Patterning-mediated supramolecular assembly of lipids into nanopalms.
At nanoconfined interfaces, a micellar ink of lipids was programmed to transform into various secondary structures such as discs, sheets, or sheet and discs via patterning-mediated self-assembly facilitated by polymer pen lithography. Nanoconfinement with printing force, humidity, temperature, pattern size, and total printing time all governed the intramolecular assembly of lipids and determined their structural shape and size. For example, disc or sheet architectures self-organized to produce cochleates or discotic liquid crystals, respectively. In contrast, the combined structure of sheet and discs produced a novel supramolecular output referred to as "nanopalms". The mechanism of nanopalms formation and the origin of their stability were investigated and discussed. Post patterning treatment helped to transform the patterned discs into ribbons and sheets into cochleates that could facilitate the curling of ribbons along their folding direction in a programmed manner via intermolecular self-organization producing the nanopalms.


Contour interaction in the presence of congenital nystagmus.
Using a computer-generated acuity task, the resolution of vertical and horizontal Landolt Cs was assessed at three levels of contrast, each for three different crowding conditions. Eighteen adults participated; six had congenital idiopathic nystagmus, six were oculocutaneous albinos and six served as controls. Contour interaction was evident when bars were placed 1 gap-width from the Landolt C and was more marked as contrast increased. When scaled to the individual resolution limit, the magnitude of contour interaction displayed by the albinos was not significantly different from the controls, however the idiopaths did exhibit a greater crowding effect than the controls. These results imply that fixation instability is a contributory factor in contour interaction, but only when retinal image velocity is above a certain threshold.


Core-shell Au@MIL-100 (Fe) as an enhanced substrate for flunixin meglumine ultra-sensitive detection.
This study aimed to develop and validate a simple and efficient surface-enhanced Raman spectroscopy (SERS) method to determine flunixin meglumine (FM) residues in animal tissues through using core-shell Au@MIL-100 (Fe) as enhanced substrate. Au@MIL-100 (Fe) composite material was synthesized by coating metal-organic framework materials (MOFs) on the surface of gold nanoparticles using the solvothermal method. Transmission electron microscopy (TEM), UV-vis spectrum, SERS spectrum, X-ray diffraction (XRD), Infrared spectrum (FT-IR), and EDX elemental mapping results revealed that the structural composition of the compound has good properties with localized surface plasmon resonance (LSPR) properties, high adsorption capacity, excellent SERS sensitivity and stability. When it was used as SERS substrate, the results of quantitative analysis of FM in pork showed a linear range of 0.10-50 mg·L-1 with a correlation coefficient (R2) of 0.9819, the limit of detection (LOD) of 0.15 mg·g-1, the recovery rate of 88.94%∼104.77%, the intra- and inter- batch relative standard deviation (RSD) of 3.57%∼14.22% and 0.18%∼3.44% respectively. Further verification results of the existing standard methods showed no significant difference between the SERS and UV methods (P < 0.05), as well as demonstrating that the SERS method has optimal precision, accuracy, and practicality. These results exposed that Au@MIL-100 (Fe) as a SERS substrate has great potential in rapid and on-site detection analysis.


Innervation of dog ciliary ganglion.
There are species differences with regard to the composition of the ciliary ganglion. For instance, in rabbits and cats it consists solely of oculomotor nerves and has no sympathetic or sensory innervation. The purpose of this study is to clarify the participation of these nerves in the ciliary ganglion of the dog by histochemical methods. Cholinesterase (ChE) activity was studied by Karnovsky's method and catecholamine fluorescence by the glyoxylic acid method. Furthermore, the origins of the respective nerves were investigated by a serial preparation method, involving unilateral cervical sympathectomy and tracer dye injection in the ganglion. The results obtained were: (1) Ciliary ganglion cells showed intense ChE activity. Oculomotor nerve fibers leading to the ganglion showed moderate ChE activity, while the reaction in the short ciliary nerves was strong. (2) Aminergic nerves were present in the intercellular space of the ciliary ganglion, and bilateral or central innervation was suggested by the results of cervical sympathectomy. (3) Connection between the ciliary and trigeminal ganglia was proved by the dye tracer study. The results show that the ciliary ganglion in dogs is composed of oculomotor, trigeminal and sympathetic nerves.


Directing two-dimensional molecular crystallization using guest templates.
The use of a coronene guest template directs the formation of a 2D Kagomé network in preference to alternative close packed and parallel hydrogen-bonded structures of tetracarboxylic acid tectons self-assembled from solution on a graphite surface.


Percutaneous cholecystostomy is safe and effective option for acute calculous cholecystitis in select group of high-risk patients.
In high-risk patients with acute calculous cholecystitis (ACC), percutaneous cholecystostomy (PC) can serve as a bridging option to cholecystectomy [laparoscopic cholecystectomy (LC)] or as definitive treatment. The purpose of this study was to identify predictors of the need for permanent PC. Data from 257 PCs performed for ACC (mean age 67.3 ± 14) was collected for a 10-year period. Demographic and clinical characteristics at initial admission, co-morbidities were analyzed. Patients who underwent interval LC were defined as the surgery group (SG; n = 163, 63.4 %) and the remaining patients as the non-SG (NSG; n = 94, 36.6 %). Patients in the SG were significantly younger and had a shorter length of hospital stay (p < 0.01). The rate of coronary artery disease (CAD; 63.2 vs. 20.2 %), chronic renal failure (14.9 vs. 6.1 %), and the mean number of co-morbidities (2.2 vs. 1.4) were significantly higher in the NSG. Sepsis at admission was more common in the NSG (19.1 vs. 4.9 %, p < 0.001). 56 patients (34.4 %) in the SG and 24 patients (25.5 %) in the NSG developed tube-related complications. In hospital mortality was similar between the groups. Multivariate analysis showed that age ≥75, increased alkaline phosphatase (ALK-P), history of CAD, were predictors of PC as a definite treatment in this high-risk group of patients with ACC. High operative risk due to older age and CAD preclude LC in more than one-third of patients following PC especially presenting with sepsis and elevated ALK-P. This study suggests that PC could be a safe treatment option in this select group of high-risk patients.


Search Methods for Systematic Reviews and Bibliographic Articles Can Improve: Responsibilities of Authorship Are Vast.
Certain types of scientific articles, including bibliographic articles, systematic reviews, and meta-analyses, require systematic search of electronic databases. Literature must be searched using clearly specified search terms, dates, and algorithms; article inclusion and exclusion criteria; and explicitly named databases. Search methods must be described in detail to allow reproducibility. In addition, responsibilities of all authors include contributing to study conception, design, data acquisition, analysis or interpretation; drafting or critically revising the work; approving the final version to be published; being accountable for the accuracy and integrity; being available to respond to queries including after publication; being able to identify which co-authors are responsible for which parts; and maintaining primary data and underpinning analysis for at least 10 years. The responsibilities of authorship are vast.


Assessment of nitric oxide metabolites concentrations in plasma, saliva, and breast milk and their relationship in lactating women.
Nitric oxide (NO) plays a role in many biological mechanisms. The amounts of physiologically produced NO are associated with the concentrations of its metabolites nitrate and nitrite. This study investigated whether there is any association between the concentrations of NO metabolites nitrate, nitrite, and nitrosylated species (RXNO) in mature breast milk, saliva, and plasma in healthy lactating women (N = 30). We hypothesized that the NO metabolites concentrations in plasma are associated with those found in saliva and in breast milk. NO metabolites concentrations were measured using chemiluminensce-based assays. Nitrate concentrations in breast milk are twice as much as plasma concentrations, whereas nitrate concentrations in saliva are about eightfold higher (both P < 0.001). Similar differences were found when nitrite concentrations were taken into consideration. RXNO concentrations in breast milk were negligible, and RXNO concentrations in saliva were approximately sixfold higher than those found in plasma samples (P < 0.0001). Nitrate concentrations in plasma are associated with nitrate concentrations in saliva (rs = 0.474, P = 0.004). However, no significant association was found between nitrate concentrations in breast milk and in plasma (P > 0.05). Our results show a significant association between nitrate concentrations in plasma with those found in saliva, whereas all other relationships were not significant. In conclusion, this report shows for the first time that the physiological concentrations of NO metabolites in human breast milk are probably independent of circulating NO metabolites concentrations and may depend mostly on endogenous NO synthesis in the breast. These findings may have clinical implications for newborns and lactating women.


Analysis of the peanut agglutinin molten globule-like intermediate by limited proteolysis.
These studies attempt to characterize the molten globule-like intermediate in the unfolding pathway of peanut agglutinin (PNA). PNA is the only known example of a homotetrameric protein that lacks the 2,2,2 or the fourfold symmetry. Previous studies have shown that PNA describes a non two-state unfolding process populated with a clearly defined intermediate. The intermediate is monomeric and has lost most of its tertiary structure and has a substantial amount of secondary structure still intact, thus described as a molten-globule (MG)-like intermediate. It was also shown by isothermal titration calorimetry to bind to lactose and some other ligands with an affinity similar to that of the native protein. This paper describes limited protease cleavage experiments on the intermediate using trypsin and protease V8 for its structural characterization. There are two hydrophobic cores in the PNA subunit. These experiments suggest that in the MG-like intermediate, the second hydrophobic core, near the sugar-binding loop of the protein loosens up. This effect is significantly reduced by the presence of 90% saturating lactose, as deduced by a reduction in cleavage propensity. This is also supported by the gain in the tertiary structure as observed by near-UV CD.


Days spent on non-invasive ventilation support: can it determine when to initiate VV- ECMO? Observational study in a cohort of Covid-19 patients.
The study evaluates the impact of the time between commencing non-invasive ventilation (NIV) support and initiation of venovenous extracorporeal membrane oxygenation (VV-ECMO) in a cohort of critically ill patients with coronavirus disease 2019 (COVID-19) associated acute respiratory distress syndrome (ARDS). Prospective observational study design in an intensive Care Unit (ICU) of a tertiary hospital in Barcelona (Spain). All patients requiring VV-ECMO support due to COVID-19 associated ARDS between March 2020 and January 2022 were analysed. Survival outcome was determined at 90 days after VV-ECMO initiation. Demographic data, comorbidities at ICU admission, RESP (respiratory ECMO survival prediction) score, antiviral and immunomodulatory treatments received, inflammatory biomarkers, the need for vasopressors, the thromboprophylaxis regimen received, and respiratory parameters including the length of intubation previous to ECMO and the length of each NIV support (high-flow nasal cannula, continuous positive airway pressure and bi-level positive airway pressure), were also collated in order to assess risk factors for day-90 mortality. The effect of the time lapse between NIV support and VV-ECMO on survival was evaluated using logistic regression and adjusting the association with all factors that were significant in the univariate analysis. Seventy-two patients finally received VV-ECMO support. At 90 days after commencing VV-ECMO 35 patients (48%) had died and 37 patients (52%) were alive. Multivariable analysis showed that at VV-ECMO initiation, age (p = 0.02), lactate (p = 0.001), and days from initiation of NIV support to starting VV-ECMO (p = 0.04) were all associated with day-90 mortality. In our small cohort of VV-ECMO patients with COVID-19 associated ARDS, the time spent between initiation of NIV support and VV-ECMO (together with age and lactate) appeared to be a better predictor of mortality than the time between intubation and VV-ECMO.


A model qualification method for mechanistic physiological QSP models to support model-informed drug development.
Mechanistic physiological modeling is a scientific method that combines available data with scientific knowledge and engineering approaches to facilitate better understanding of biological systems, improve decision-making, reduce risk, and increase efficiency in drug discovery and development. It is a type of quantitative systems pharmacology (QSP) approach that places drug-specific properties in the context of disease biology. This tutorial provides a broadly applicable model qualification method (MQM) to ensure that mechanistic physiological models are fit for their intended purposes.


Zinc metabolism and immune function in healthy elderly people.
Changes in immune function have been described in the elderly, and zinc is known to influence immune parameters. Zinc supplementation has, however, produced contradictory results. We evaluated zinc metabolism and immunity in 15 healthy non-institutionalised elderly people on a self-selected diet and compared the results with those obtained from 10 young healthy subjects. The elderly subjects had reduced serum zinc levels and their zinc absorption values were lower than in the young subjects. Circulating immune parameters were within the normal range, while delayed immune response was altered in 8 of the 15 elderly subjects. Zinc supplementation (25 mg/day) did not result in a significant improvement in dermal hypersensitivity responses although some effect was seen in 5 cases. It is concluded that zinc supplementation is unnecessary in elderly healthy subjects even if they present zinc metabolism alterations, but may be indicated in the presence of disease.


[Synovial chondrosis. Chondromatosis--osteochondromatosis].
Clinical and pathological findings as well as diagnosis of Chondrosis synovialis (Henderson-Jones Syndrome) are presented and completed with authors observations. According to relevant literature the treatment should depend on the stage of the disease.


An N-terminal SIAH-interacting motif regulates the stability of the ubiquitin specific protease (USP)-19.
The Ubiquitin Specific Protease-19 (USP19) regulates cell cycle progression and is involved in the cellular response to different types of stress, including the unfolded protein response (UPR), hypoxia and muscle atrophy. Using the unique N-terminal domain as bait in a yeast-two hybrid screen we have identified the ubiquitin ligases Seven In Absentia Homolog (SIAH)-1 and SIAH2 as binding partners of USP19. The interaction is mediated by a SIAH-consensus binding motif and promotes USP19 ubiquitylation and proteasome-dependent degradation. These findings identify USP19 as a common substrate of the SIAH ubiquitin ligases.


Chromatographic resolution of atropisomers for toxicity and biotransformation studies in pharmaceutical research.
Atropisomerism can be a complex concept for those who have not encountered it before. This paper discusses the experiments for identification, isolation, thermal stability, toxicity and biotransformation of various species. The identified atropisomers are a series of rotational hindered biaryl, rotational hindered amide, ring flip, and macrocycles atropisomers identified using supercritical fluid chromatography (SFC) and high performance liquid chromatography (HPLC). These technologies offered the advantage of separating various atropoenantiomers, atropdiastereomers and mixed atropisomers with other forms of stereoisomers in both analytical and preparative scales. With ultra-performance convergence chromatography (UPC(2)), the detection of N-oxide atropisomer metabolites can be obtained at very low level thus enabling the observation of conversion in human plasma possible. As the resolution of atropisomers are related to the energy barriers on the rotational axis, a calculated computational protocol was developed to predict the formation. A threshold of 10kcal/mol was established for possible detection of the atropisomers' existence with chromatographic technologies at room temperature or above. The atropisomer with higher energy barrier (>20kcal/mol) were isolated via preparative chromatography and the isolates studied in vitro and in vivo for evaluation of their stability in human plasma. The detailed analytical method development to analyze the biotransformation of the atropisomers in human plasma are also discussed in this paper.


Colostomy fistula caused by segmental absence of intestinal musculature: a case report.
Segmental Absence of Intestinal Musculature (SAIM) is a rare condition associated with intestinal obstruction and perforation. Colostomy fistula as a presentation of SAIM and their association with anastomotic failure have not been previously reported. This case report aimed to raise awareness of this unique manifestation and its potential implications. A 58-year-old male with a history of type 2 diabetes, hypertension, and lumbar hernia presented with diarrhea. Lower gastrointestinal endoscopy revealed a tumor in the rectum, for which he was diagnosed with a well-differentiated adenocarcinoma. The patient underwent a laparoscopic Hartmann operation. After the operation, an entero-entero-fistula was identified at the sigmoid colostomy site. Subsequently, laparoscopic reconstruction of the colostomy was performed, and the patient had a favorable postoperative course without complications. Histopathological examination confirmed the localized absence of the muscularis propria in the resected colon, with fibrosis and nearby ganglion cells. This case highlights the rarity of a colostomy fistula as a manifestation of SAIM and emphasizes the need to consider SAIM in the differential diagnosis for such cases. The presence of SAIM-affected lesions poses a risk of anastomotic failure, underscoring the importance of assessing the risk of complications during future surgeries. Surgeons should be aware of the etiology and potential implications of SAIM to ensure appropriate management and minimize postoperative morbidity. Further studies are warranted to explore the underlying mechanisms and optimize surgical strategies for patients with SAIM and its associated complications. Increased awareness among clinicians is crucial for timely diagnoses and tailored interventions to improve patient outcomes.


Psychometric testing of the Persian version of the Nursing Care Quality Scale: A methodological study.
To evaluate the psychometric properties of the Persian version of the Cambodian Nursing Care Quality Scale. Methodological design. This study was carried out in several steps: a forward-backward translation was done, face and construct validity was measured using exploratory and confirmatory factor analysis and the reliability was evaluated. A convenience sampling method was employed to recruit 350 nurses from May 2021 to March 2022. The exploratory factor analysis extracted six factors that explained 60.76% of the total variance. The six-factor model is supported by confirmatory factor analysis. The Cronbach's alpha and the intra-class correlation coefficient were 0.94 and 0.85, respectively. Evaluation of the quality of care can lead to the enhancement of the quality of nursing services and patient safety. This will subsequently increase the patients' and community satisfaction.


Cerebrospinal Fluid Polymerase Chain Reaction in the Diagnosis of Neonatal Bacterial Meningitis: A Single-Center Experience From Vietnam.
To compare the performance of cerebrospinal fluid (CSF) polymerase chain reaction (PCR) with bacterial culture for the diagnosis of neonatal bacterial meningitis (NBM). The CSF analysis of neonate with confirmed bacterial meningitis was performed with PCR and bacterial culture, and results were compared. Among 24 neonates, the pathogens Identified included E. coli K1, GBS, Streptococcus pneumoniae and Listeria. PCR identified 20 (83.3%) pathogens, and culture 4 (16.7%) pathogens. Prior antibiotics were administered to 20 (83.3%) neonates in whom PCR identified 17 (85%) and culture 3 (15%) pathogens. CSF PCR had a higher yield of pathogens than CSF culture in confirmed neonatal bacterial meningitis with a high rate of prior antibiotic therapy.


Low-Dose Ticagrelor in Patients With High Ischemic Risk and Previous Myocardial Infarction: A Multicenter Prospective Real-World Observational Study.
Prolonged dual antiplatelet therapy after 12 months in patients with previous myocardial infarction (MI) is attractive to reduce long-term ischemic complications. In the PEGASUS-TIMI 54, the use of low-dose ticagrelor (60 mg b.i.d.) plus aspirin after 12 months from MI reduced the risk of ischemic events, at the price of limited increase on bleeding complications. However, data on the use of low-dose ticagrelor in real-world practice lack. We aim at providing data on prescription/eligibility criteria and outcomes in patients receiving low-dose ticagrelor in the real-world setting. We enrolled consecutive patients eligible for ticagrelor 60 mg according to Italian national regulation in 3 high-volume centers and collected 1-year outcomes. The primary objective of the study is to generate real-world data about clinical characteristics, eligibility criteria, major adverse cardiovascular events, bleeding, and adverse event in patients receiving low-dose ticagrelor from our cohort. One hundred eighty-one patients were consecutively enrolled with a median follow-up of 18 months. The most used and the least used prescription criteria were multivessel coronary disease (72.4%) and chronic kidney disease (15.5%), respectively. At 1-year follow-up, the rate of major adverse cardiovascular events was 4.97%; of these, 3.86% of patients had a MI, and 1.1% had a stroke/transient ischemic attack, whereas no major bleeding occurred. In conclusion, in a real-world study, including patients with previous MI, low-dose ticagrelor for prolonged dual antiplatelet therapy showed to be effective and safe, with no major bleeding occurring at follow-up.


SPECT/CT of Total Ankle Arthroplasty.
Use of SPECT/CT (Single Photon Emission Computed Tomography/Computed Tomography) is increasing providing additional information in patients with inconclusive clinical examination and unremarkable imaging findings presenting with chronic pain after total ankle arthroplasty. To differentiate the cause of pain after total ankle arthroplasty can be challenging. SPECT/CT combines structural and metabolic imaging as a hybrid tool leading to higher specificity and overall diagnostic accuracy presumably in cases of gutter impingement, prosthetic loosening, and osteoarthritis of adjacent joints. Moreover, SPECT/CT can complement diagnostic work up in periprosthetic joint infections. Basal tracer enhancement has to be considered for the interpretation of imaging findings.


Correction: Do honey bee (Apis mellifera) foragers recruit their nestmates to native forbs in reconstructed prairie habitats?
[This corrects the article DOI: 10.1371/journal.pone.0228169.].


Encapsulation and release of living tumor cells using hydrogels with the hybridization chain reaction.
Circulating tumor cells (CTCs) enable noninvasive liquid biopsy and identification of cancer. Various approaches exist for the capture and release of CTCs, including microfluidic methods and those involving magnetic beads or nanostructured solid interfaces. However, the concomitant cell damage and fragmentation that often occur during capture make it difficult to extensively characterize and analyze living CTCs. Here, we describe an aptamer-trigger-clamped hybridization chain reaction (atcHCR) method for the capture of CTCs by porous 3D DNA hydrogels. The 3D environment of the DNA networks minimizes cell damage, and the CTCs can subsequently be released for live-cell analysis. In this protocol, initiator DNAs with aptamer-toehold biblocks specifically bind to the epithelial cell adhesion molecule (EpCAM) on the surface of CTCs, which triggers the atcHCR and the formation of a DNA hydrogel. The DNA hydrogel cloaks the CTCs, facilitating quantification with minimal cell damage. This method can be used to quantitively identify as few as 10 MCF-7 cells in a 2-µL blood sample. Decloaking of tumor cells via gentle chemical stimulus (ATP) is used to release living tumor cells for subsequent cell culture and live-cell analysis. We also describe how to use the protocol to encapsulate and release cells of cancer cell lines, which can be used in preliminary experiments to model CTCs. The whole protocol takes ~2.5 d to complete, including downstream cell culture and analysis.


Antiproliferative and ultrastructural effects of BPQ-OH, a specific inhibitor of squalene synthase, on Leishmania amazonensis.
Parasites of the Leishmania genus require for the growth and viability the de novo synthesis of specific sterols as such as episterol and 5-dehydroepisterol because cholesterol, which is abundant in their mammalian hosts, does not fulfill the parasite sterol requirements. Squalene synthase catalyzes the first committed step in the sterol biosynthesis and has been studied as a possible target for the treatment of high cholesterol levels in humans. In this work we investigated the antiproliferative and ultrastructural effects induced by 3-(biphenyl-4-yl)-3-hydroxyquinuclidine (BPQ-OH), a specific inhibitor of squalene synthase, on promastigote and amastigote forms of Leishmania amazonensis. BPQ-OH had a potent dose-dependent growth inhibitory effect against promastigotes and amastigotes, with IC(50) values 0.85 and 0.11 microM, respectively. Ultrastructural analysis of the treated parasites revealed several changes in the morphology of promastigote forms. The main ultrastructural change was found in the plasma membrane, which showed signs of disorganization, with the concomitant formation of elaborated structures. We also observed alterations in the mitochondrion-kinetoplast complex such as mitochondrial swelling, rupture of its internal membrane and an abnormal compaction of the kinetoplast. Other alterations included the appearance of multivesicular bodies, myelin-like figures, alterations of the flagellar membrane and presence of parasites with two or more nuclei and kinetoplasts. We conclude that the BPQ-OH was a potent growth inhibitor of L. amazonensis, which led to profound changes of the parasite's ultrastructure and might be a valuable lead compound for the development of novel anti-Leishmania agents.


Shape complementarity, binding-site dynamics, and transition state stabilization: a theoretical study of Diels-Alder catalysis by antibody 1E9.
Antibody 1E9 is a protein catalyst for the Diels-Alder reaction between tetrachlorothiophene dioxide and N-ethylmaleimide. Quantum mechanical calculations have been employed to study the 1E9-catalyzed Diels-Alder reaction in the gas phase. The transition states and intermediates were all determined at the B3LYP/6-31G*//HF/6-31G* level. The cycloaddition step is predicted to be rate-determining, and the endo reaction pathway is strongly favored. Binding of the reactants and the transition states to antibody 1E9 was investigated by docking and molecular dynamics simulations. The linear interaction energy (LIE) method was adopted to estimate the free energy barrier of the 1E9-catalyzed Diels-Alder reaction. The catalytic efficiency of antibody 1E9 is achieved by enthalpic stabilization of the transition state, near-perfect shape complementarity of the hydrophobic binding site for the transition state, and a strategically placed hydrogen-bonding interaction.


Transgender Adolescents' Uses of Social Media for Social Support.
Transgender adolescents are at higher risk for negative mental health outcomes, such as depression, anxiety, and suicidality, compared with cisgender adolescents. Social media may be an effective venue for addressing these health disparities because most adolescents have access to online information and socialization. This study used qualitative inquiry to explore transgender adolescents' uses of social media for social support. Transgender adolescents between the ages of 15 and 18 years with a social media profile were recruited from a pediatric gender clinic in the Midwestern U.S. A 30- to 60-minute semistructured interview assessed ways participants used social media to access transgender-related support. Thematic analysis was used to develop categories and code the transcripts. Coding discrepancies were resolved by two researchers following initial coding. Results represent data from 25 interviews comprising 13 transmasculine, 11 transfeminine, and one nonbinary participant(s), with 68% of the sample identifying as white, non-Hispanic. Categories emerged regarding forms of support participants received from transgender-related online communities including emotional support through peers and role models, appraisal support for validating their experiences, and informational support for navigating health decisions and educating family and friends. Participants also referenced negative experiences, including harassment and exclusionary behavior online. Social media platforms represent hubs of community for transgender adolescents. These communities provide emotional, appraisal, and informational support that transgender youth may not otherwise be able to access. Future research should use the affordances of social media to identify approaches to addressing health disparities and improving the well-being of transgender adolescents.


Building an improved transcription factor-centered yeast one hybrid system to identify DNA motifs bound by protein comprehensively.
Identification of the motifs bound by a transcription factor (TF) is important to reveal the function of TF. Previously, we built a transcription factor centered yeast one hybrid (TF-Centered Y1H) that could identify the motifs bound by a target TF. However, that method was difficult to comprehensively identify all the motifs bound by a TF. Here, we build an improved TF-Centered Y1H to comprehensively determine the motifs bound by a target TF. Recombination-mediated cloning in yeast was performed to construct a saturated prey library that contains 7 random base insertions. After TF-Centered Y1H screening, all the positive clones were pooled together to isolate pHIS2 vector. The insertion regions of pHIS2 were PCR amplified and the PCR product was subjected to high-throughput sequencing. The insertion sequences were then retrieved and analyzed using MEME program to identify the potential motifs bound by the TF. Using this technology, we studied the motifs bound by an ethylene-responsive factor (BpERF2) from birch. In total, 22 conserved motifs were identified, and most of them are novel cis-acting elements. Both the yeast one hybrid and electrophoretic mobility shift assay verified that the obtained motifs could be bound by BpERF2. In addition, chromatin immunoprecipitation (ChIP) study further suggested that the identified motifs can be bound by BpERF2 in cells of birch. These results together suggested that this technology is reliable and has biological significance. This method will have wide application in DNA-protein interaction studies.


Hypermutability and mismatch repair deficiency in RER+ tumor cells.
A subset of sporadic colorectal tumors and most tumors developing in hereditary nonpolyposis colorectal cancer patients display frequent alterations in microsatellite sequences. Such tumors have been thought to manifest replication errors (RER+), but the basis for the alterations has remained conjectural. We demonstrate that the mutation rate of (CA)n repeats in RER+ tumor cells is at least 100-fold that in RER- tumor cells and show by in vitro assay that increased mutability of RER+ cells is associated with a profound defect in strand-specific mismatch repair. This deficiency was observed with microsatellite heteroduplexes as well as with heteroduplexes containing single base-base mismatches and affected an early step in the repair pathway. Thus, a true mutator phenotype exists in a subset of tumor cells, the responsible defect is likely to cause transitions and transversions in addition to microsatellite alterations, and a biochemical basis for this phenotype has been identified.


Absence of immunoglobulin and T-cell receptor gene rearrangements in myelodysplastic syndromes and acute nonlymphocytic leukemias.
The arrangement of the immunoglobulin and T-cell receptor genes has been analysed in 72 cases of primary myelodysplastic syndrome and 17 cases of acute nonlymphocytic leukemia. DNA was extracted from bone marrow aspirates, digested with at least two restriction enzymes, and hybridised with probes for the joining region of the immunoglobulin heavy chain gene, the constant region of the T-cell receptor beta chain gene, and the joining region of the T-cell receptor gamma chain gene. All cases showed germline arrangements of the immunoglobulin and the T cell receptor genes. Thus, true interlineage infidelity, myeloid to lymphoid, is a rare occurrence in myelodysplasia and in myeloid leukemias.


A 'catch and release' strategy towards HPLC-free purification of synthetic oligonucleotides by a combination of the strain-promoted alkyne-azide cycloaddition and the photocleavage.
A convenient strategy to purify oligonucleotides (ONs) synthesized by solid phase synthesis on an automatic DNA/RNA synthesizer was described. By attaching a photocleavable azide linker as the last phosphoramidite unit in the ON synthesis, only the desired full-length sequence was 'caught' on a controlled pore glass (CPG) resin possessing an aza-dimethoxycyclooctyne (DIBAC) derivative. Washing the resulting CPG resin to remove all unbounded species, the subsequent photoirradiation allowed the pure ONs to be 'released' without leaving any chemical modifications on native ON structure or chemical reagents from the solid phase ON synthesis.


Indirect impact of Covid-19 on hospital care pathways in Italy.
Earlier in 2020, seven Italian regions, which cover 62% of the Italian population, set up the Mimico-19 network to monitor the side effects of the restrictive measures against Covid-19 on volumes and quality of care. To this aim, we retrospectively analysed hospital discharges data, computing twelve indicators of volume and performance in three clinical areas: cardiology, oncology, and orthopaedics. Weekly indicators for the period January-July 2020 were compared with the corresponding average for 2018-2019; comparisons were performed within 3 sub-periods: pre-lockdown, lockdown, and post-lockdown. The weekly trend of hospitalisations for ST-segment elevation myocardial infarction (STEMI) showed a 40% reduction, but the proportion of STEMI patients with a primary PTCA did not significantly change from previous years. Malignant neoplasms surgery volumes differed substantially by site, with a limited reduction for lung cancer (< 20%) and greater declines (30-40%) for breast and prostate cancers. The percentage of timely surgery for femoral neck in the elderly remained constantly higher than the previous 2 years whereas hip and knee replacements fell dramatically. Hospitalisations have generally decreased, but the capacity of a timely and effective response in time-dependent pathways of care was not jeopardized throughout the period. General trends did not show important differences across regions, regardless of the different burden of Covid-19. Preventive and primary care services should adopt a pro-active approach, moving towards the identification of at-risk conditions that were neglected during the pandemic and timely addressing patients to the secondary care system.


[Analysis of EBV-LMP1 gene mutation in plasma of patients with nasopharyngeal carcinoma in Chongqing].
To detect and analyze the EBV-LMP1 gene N terminus XhoI-site mutation and C terminus 30 bp deletion in plasma of patients with nasopharyngeal carcinoma (NPC) in Chongqing of China. DNA extraction and PCR amplification was used in plasma of 48 NPC patients and 40 control non-NPC cases from Chongqing of China. All the PCR production of N terminus was digested by enzyme XhoI, then segregated in 8% PAGE. All- the PCR production of C terminus was segregated in 8% PAGE too. Compared with B95-8 cell, some of the PCR production were sequenced and analyzed with software. LMP1 was amplified successfully from 48 of 48 NPC cases (100%) and from 38 of 40 non-NPC cases (95%) by PCR, but none of LMP1 XhoI-site mutation and none of 30 bp deletion was found by digesting, segregating and Sequencing. EBV-LMP1 gene N terminus XhoI-site and C terminus 30 bp deletion have no mutation in plasma of NPC or non-NPC cases in Chongqing of China. The precise relationship of EBV-LMP1 gene mutation or deletion with NPC pathogenesis needs further investigation.


Novel opportunities from bioimaging to understand the trafficking and maturation of intracellular pulmonary surfactant and its role in lung diseases.
Pulmonary surfactant (PS), a complex mixture of lipids and proteins, is essential for maintaining proper lung function. It reduces surface tension in the alveoli, preventing collapse during expiration and facilitating re-expansion during inspiration. Additionally, PS has crucial roles in the respiratory system's innate defense and immune regulation. Dysfunction of PS contributes to various respiratory diseases, including neonatal respiratory distress syndrome (NRDS), adult respiratory distress syndrome (ARDS), COVID-19-associated ARDS, and ventilator-induced lung injury (VILI), among others. Furthermore, PS alterations play a significant role in chronic lung diseases such as chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF). The intracellular stage involves storing and releasing a specialized subcellular organelle known as lamellar bodies (LB). The maturation of these organelles requires coordinated signaling to organize their intracellular organization in time and space. LB's intracellular maturation involves the lipid composition and critical processing of surfactant proteins to achieve proper functionality. Over a decade ago, the supramolecular organization of lamellar bodies was studied using electron microscopy. In recent years, novel bioimaging tools combining spectroscopy and microscopy have been utilized to investigate the in cellulo intracellular organization of lamellar bodies temporally and spatially. This short review provides an up-to-date understanding of intracellular LBs. Hyperspectral imaging and phasor analysis have allowed identifying specific transitions in LB's hydration, providing insights into their membrane dynamics and structure. A discussion and overview of the latest approaches that have contributed to a new comprehension of the trafficking and structure of lamellar bodies is presented.


Evaluation of the clinical performance of the HISCL-5000 analyzer in the detection of Krebs von den Lungen-6 antigen and its diagnostic value in interstitial lung disease.
The sputum saccharide chain antigen (Krebs von den Lungen-6 [KL-6]) is a serum biomarker of lung injury. We aimed to evaluate the clinical performance of the automated immunoassay analyzer HISCL-5000 in detecting KL-6 by comparing it with LUMIPULSE G1200 and determine the diagnostic value of KL-6 in interstitial lung disease (ILD). A total of 145 serum samples from patients were tested using the two automated immunoassay analyzers in parallel. With a cutoff level of 500 U/mL, comparing the two systems, the agreement, sensitivity, specificity, and kappa value were 99.20%, 100%, 98.63%, and 0.984 (95% CI, 0.952-1.000), respectively. Spearman's correlation and ICC showed that there was a strong correlation between serum KL-6 levels measured by the two systems (rS = .991 [95% CI, 0.981-0.995], ICC = 0.984 [95% CI, 0.978-0.989], P < .01). The clinical diagnosis agreement rate in both systems was >80%. The kappa value was 0.707 (95% CI, 0.582-0.832; SYSTEM B) and 0.707 (95% CI, 0.588-0.826; SYSTEM A). The KL-6 level in the ILD group (1339.5, 662.5-2363) was significantly higher than that in the non-ILD groups (252, 158.5-353; Mann-Whitney U = 381.5, P < .01), and the KL-6 level (1558, 726-2772.5) in the ILD group detected by SYSTEM A was significantly higher than that in the lung cancer group (339, 207-424), other respiratory disease group (249, 194-366), and control group (198, 131.5-297; Kruskal-Wallis H = 63.19, P < .01). HISCL-5000 showed well-concordant results with those of HISCL-5000 in the KL-6 tests. In patients with ILD, KL-6 showed a good diagnostic performance.


Traumatic spinal cord injury in Tianjin, China: a single-center report of 354 cases.
Hospital-based retrospective study. The objective of this study was to describe the epidemiological profile of traumatic spinal cord injury (TSCI) in Tianjin Medical University General Hospital, China, from 2009 to 2014. Tianjin Medical University General Hospital. Hospital medical records of patients with TSCI admitted to hospital from 1 January 2009 to 31 December 2014 were reviewed. Collected variables included gender, age, marital status, ethnic group, occupation, etiology, neurological level of injury, American Spinal Injury Association (ASIA)-ISCoS impairment scale at admission, the severity, death and its cause, concomitant injuries and treatment choice. During the study period, 354 cases were identified. Male-to-female ratio was 2.34:1, with a mean age of 50.1±15.5 years. Falls (55.1%), comprising low falls and high falls (33.6% and 21.5%, respectively), were the leading cause, followed by motor vehicle collisions (MVCs) (35.9%). The most common injury site was the cervical spinal cord, especially C4-C6, accounting for 59.3%. Surgery was the major treatment choice (57.6%). The number of TSCI patients increased annually in our center. The mean age at the time of injury was older, and the proportion of males was higher. The leading two causes were falls and MVCs. The SCIs caused by MVCs were increasing. Peasants, workers and unemployed individuals were those at higher risk. Surgery was the major treatment choice. These data may be useful to implement those preventive strategies focused on the characteristics of different groups and pay more attention to high-risk populations.


Hepatitis B virus envelope proteins can serve as therapeutic targets embedded in the host cell plasma membrane.
Hepatitis B virus (HBV) infection is a major health threat causing 880,000 deaths each year. Available therapies control viral replication but do not cure HBV, leaving patients at risk to develop hepatocellular carcinoma. Here, we show that HBV envelope proteins (HBs)-besides their integration into endosomal membranes-become embedded in the plasma membrane where they can be targeted by redirected T-cells. HBs was detected on the surface of HBV-infected cells, in livers of mice replicating HBV and in HBV-induced hepatocellular carcinoma. Staining with HBs-specific recombinant antibody MoMab recognising a conformational epitope indicated that membrane-associated HBs remains correctly folded in HBV-replicating cells in cell culture and in livers of HBV-transgenic mice in vivo. MoMab coated onto superparamagnetic iron oxide nanoparticles allowed to detect membrane-associated HBs after HBV infection by electron microscopy in distinct stretches of the hepatocyte plasma membrane. Last but not least, we demonstrate that HBs located on the cell surface allow therapeutic targeting of HBV-positive cells by T-cells either engrafted with a chimeric antigen receptor or redirected by bispecific, T-cell engager antibodies. TAKE AWAYS: HBs become translocated to the plasma membrane. Novel, recombinant antibody confirmed proper conformation of HBs on the membrane. HBs provide an interesting target by T-cell-based, potentially curative therapies.


Exstrophic bladder duplication in the sagittal plane: Surgical management of a rare case.
The bladder-Exstrophy-Epispadias complex (BEEC) contains a wide spectrum of congenital malformations. A treatment naïve refugee was referred to our center with what was identified as BEEC. A 27-year-old female patient was referred for total incontinence since birth by the general practitioner from the refugee center. An exstrophic bladder with blind ending ureteral orifices and a second non-exstrophic bladder with two orthotopic ureters was identified, demonstrating the bladder duplication in the sagittal plane. Laparotomy was performed, dissecting the exstrophic bladder plate caudally and using it as a ventral onlay to augment the non-exstrophic bladder. A Mitchell-type bladder neck reconstruction was performed with an autologous fascia sling around the bladder neck to obtain continence. As the patient had never voluntarily voided, chances of spontaneous voiding after surgery were low. Therefore creation of a continent Mitrofanoff-type vesicostomy was additionally realized and genital reconstruction was achieved. 12 months post operatively, the patient was completely continent, had a bladder capacity of 250 ml, and performed self-catheterization 5 times a day. No post-operative complications were observed. Admission of political refugees can implicate challenging surgeries for congenital malformations in adults, such as BEEC. This demonstrates the importance of multidisciplinary transitional care.


Non-invasive monitoring of chronic liver disease via near-infrared and shortwave-infrared imaging of endogenous lipofuscin.
Monitoring the progression of non-alcoholic fatty liver disease is hindered by a lack of suitable non-invasive imaging methods. Here, we show that the endogenous pigment lipofuscin displays strong near-infrared and shortwave-infrared fluorescence when excited at 808 nm, enabling label-free imaging of liver injury in mice and the discrimination of pathological processes from normal liver processes with high specificity and sensitivity. We also show that the near-infrared and shortwave-infrared fluorescence of lipofuscin can be used to monitor the progression and regression of liver necroinflammation and fibrosis in mouse models of non-alcoholic fatty liver disease and advanced fibrosis, as well as to detect non-alcoholic steatohepatitis and cirrhosis in biopsied samples of human liver tissue.


β-Galactosidases: A great tool for synthesizing galactose-containing carbohydrates.
β-Galactosidases, an important class of glycosidases, naturally catalyze the hydrolysis of β-galactosidic bonds in oligosaccharides and polysaccharides. Traditionally, these enzymes have been used to degrade lactose in dairy products, which are beneficial for lactose-intolerant people. Attractively, β-galactosidases exhibit glycosyl transfer activity under certain conditions in vitro. They are capable of synthesizing carbohydrates from cheap starting substrates in a facile, efficient, and environment-friendly way. The condensation of lactose into the well-known prebiotic galacto-oligosaccharides by β-galactosidases has become a key aspect of the industrial interest in the synthetic activity in recent years. At present, the transglycosylation activity of these enzymes has been greatly extended. It can be used not only in building glycan blocks of crucial glycoconjugates to elucidate their biological functions, but also in glycosylation of vital molecules, which have been applied in food, medicine and cosmetic industries to improve solubility, stability and bioactivity. Further molecular engineering of β-galactosidases has significantly improved their synthetic activity, expanded the substrate spectrum and made them more powerful in carbohydrate synthesis. This review covers the classification, structure and mechanism of β-galactosidases, galactosylation reactions catalyzed by these enzymes, and various strategies of enzyme engineering, with an emphasis on recent advances.


Live-cell imaging reveals redox metabolic reprogramming during zygotic genome activation.
Metabolic programming is deeply intertwined with early embryonic development including zygotic genome activation (ZGA), the polarization of zygotic cells, and cell fate commitment. It is crucial to establish a noninvasive imaging technology that spatiotemporally illuminates the cellular metabolism pathways in embryos to track developmental metabolism in situ. In this study, we used two high-quality genetically encoded fluorescent biosensors, SoNar for NADH/NAD+ and iNap1 for NADPH, to characterize the dynamic regulation of energy metabolism and redox homeostasis during early zygotic cleavage. Our imaging results showed that NADH/NAD+ levels decreased from the early to the late two-cell stage, whereas the levels of the reducing equivalent NADPH increased. Mechanistically, transcriptome profiling suggested that during the two-cell stage, zygotic cells downregulated the expression of genes involved in glucose uptake and glycolysis, and upregulated the expression of genes for pyruvate metabolism in mitochondria and oxidative phosphorylation, with a decline in the expression of two peroxiredoxin genes, Prdx1 and Prdx2. Collectively, with the establishment of in situ metabolic monitoring technology, our study revealed the programming of redox metabolism during ZGA.


Viscerotopic control of regional vascular beds by discrete groups of neurons within the midbrain periaqueductal gray.
It is well established that a group of bulbospinal neurons within the rostral ventrolateral medulla plays a crucial role in the tonic and phasic control of arterial pressure. In the cat, these neurons are confined to a discrete region which has been termed the subretrofacial (SRF) nucleus. Recent evidence suggests that this nucleus is viscerotopically organized with respect to its control over different vascular beds. These observations raise the question as to whether functionally different subgroups of SRF pressor neurons receive inputs from supramedullary cell groups that also exert a specific control over particular vascular beds. To answer this question retrogradely transported tracers (i.e. rhodamine or fluorescein-labelled microspheres, wheat germ agglutinin-horseradish peroxidase) were injected into physiologically identified sites within the rostral or caudal parts of the SRF nucleus of the cat. Separate groups of neurons in the midbrain periaqueductal gray region (PAG) were found to project specifically to subgroups of cells within the rostral and caudal parts of the SRF nucleus. These findings, together with the results of recent functional studies of the PAG suggest that these distinct projections from the PAG to the SRF nucleus are involved in the expression of different patterns of emotionally coupled cardiovascular responses.


Role of the blue mechanism in wavelength discrimination.
The role of blue cones as well as the pathways they supply (collectively called the "blue mechanism") is evaluated by comparing ordinary wavelength discrimination functions with those obtained using two methods designed to inhibit the blue mechanism selectively. These methods use a just-noticeable-border criterion (JNB), instead of the usual one of just-noticeable-difference, and a yellow preadapting field to induce transient tritanopia. Without transient tritanopia, the data obtained using the just-noticeable-border criterion reveal a small contribution of the blue mechanism to wavelength discrimination. Transient tritanopia, with JNB, produces an additional selective loss of wavelength discrimination in a spectral region flanking 460 nm, which yields a function resembling those for tritanopes previously examined.


Integrated approach toward the discovery of glyco-biomarkers of inflammation-related diseases.
Glycobiology has contributed tremendously to the discovery and characterization of cancer-related biomarkers containing glycans (i.e., glyco-biomarkers) and a more detailed understanding of cancer biology. It is now recognized that most chronic diseases involve some elements of chronic inflammation; these include cancer, Alzheimer's disease, and metabolic syndrome (including consequential diabetes mellitus and cardiovascular diseases). By extending the knowledge and experience of the glycobiology community regarding cancer biomarker discovery, we should be able to contribute to the discovery of diagnostic/prognostic glyco-biomarkers of other chronic diseases that involve chronic inflammation. Future integration of large-scale "omics"-type data (e.g., genomics, epigenomics, transcriptomics, proteomics, and glycomics) with computational model building, or a systems glycobiology approach, will facilitate such efforts.


Five percent human albumin in lactated Ringer's solution for resuscitation from hemorrhagic shock: efficacy and cardiopulmonary consequences.
Using an ovine model of acute hemorrhagic shock, we evaluated the utility of 5% albumin in lactated Ringer's (5% ALR) solution as a resuscitation solution. After instrumentation and obtaining baseline values for BP, mean arterial pressure (MAP), pulmonary capillary wedge pressure (WP), CVP, cardiac output, extravascular lung water (EVLW), and blood gases (mixed venous and arterial), animals were rapidly exsanguinated to an MAP of 50 mm Hg. After 30 min at this pressure, measurements were repeated and 5% ALR was administered until two of three variables (WP, MAP, cardiac output) were restored to baseline values. The administration of 5% ALR was continued as needed to maintain baseline values of these variables. Sixty minutes later, data were again recorded. For induction of shock, 15.7 +/- 5.2 ml of blood/kg body weight was removed. Pulmonary artery pressure, WP, MAP, and cardiac output all significantly decreased with shock. After resuscitation, all values except MAP returned to baseline. The resuscitation volume of 5% ALR was 25.2 +/- 18.4 ml/kg. There were no changes in EVLW or intrapulmonary shunt. Oxygen delivery was significantly compromised during shock but returned to baseline after resuscitation. We conclude that in a model such as ours, 5% ALR can reverse the hemodynamic effects of acute hemorrhagic shock.


Blood eosinophilic relative count is prognostic for breast cancer and associated with the presence of tumor at diagnosis and at time of relapse.
Cancer outcome is associated with circulating immune cells, including eosinophils. Here we analyze the relative eosinophil count (REC) in different breast cancer subtypes. Stage I-III breast cancer patients were included in the study and classified as REC-high vs low (cutoff 1.5%) or relative lymphocyte count (RLC)-high vs low (cutoff 17.5%). The co-primary endpoints were the breast cancer-specific survival (BCSS) or the time to treatment failure (TTF) in the REC groups. Overall 930 patients were included in the study. We observed a benefit for REC-high vs REC-low in TTF (HR 0.610, 95% CI 0.458-0.812), and in BCSS (HR 0.632, 95% CI 0.433-0.923). Similarly, we observed a better TTF (HR 0.421, 95% CI 0.262-0.677) and BCSS (HR 0.350, 95% CI 0.200-0.614) in RLC-high vs low. A lower relapse rate was observed in the REC-high vs REC-low group (17.1% vs 24.7%, p = 0.005), not confirmed in the multivariate analysis. A lower median REC at baseline and at relapse was observed compared to REC after surgery and during cancer-free follow-up (p < .0001). REC could be a new promising, affordable and accessible predictive and prognostic biomarker in all breast cancer subtypes.


Implementing CQI projects in hospitals.
Quality improvement approaches such as continuous quality improvement (CQI) and total quality management are widely used, but little is known about how much it costs to use the principles and techniques required to implement CQI processes. In the Robert Wood Johnson Foundation's Improving the Quality of Hospital Care (IQHC) program, four consortia of hospitals were funded in the early 1990s. Interviews with quality managers at 38 of the consortia hospitals were conducted in 1995 to determine the costs of conducting CQI projects to allow an estimation of the marginal cost of using CQI processes (particularly cross-disciplinary teams) to improve quality of care. Quality managers described 69% of project outcomes as critical to clinical services. Team members identified the issues their teams addressed and selected the project 64% of the time, the methods of analysis 87% of the time, and the approaches to resolving the problem or issue 97% of the time. Most of the respondents agreed that the team members had the authority to resolve the problem without appealing to higher levels of management. Costs for hospitals' most recently completed projects varied widely, from $148 for the entire project to $18,590. The length or duration of the projects also varied widely, from 1 month to 66 months. In the hospitals included in this sample, all of which were highly self-selected (evidenced by their participation in a voluntary consortium of hospitals focused on quality of care), knowledge of CQI processes appeared to be fairly thorough. Teams appeared to have a reasonable amount of autonomy. New CQI projects should be subjected to scrutiny in terms of their likely contribution to quality of care, as distinct from other positive outcomes.


Hearing in Noise Test Brazil: standardization for young adults with normal hearing.
Individuals with the same ability of speech recognition in quiet can have extremely different results in noisy environments. To standardize speech perception in adults with normal hearing in the free field using the Brazilian Hearing in Noise Test. Contemporary, cross-sectional cohort study. 79 adults with normal hearing and without cognitive impairment participated in the study. Lists of Hearing in Noise Test sentences were randomly in quiet, noise front, noise right, and noise left. There were no significant differences between right and left ears at all frequencies tested (paired t-1 test). Nor were significant differences observed when comparing gender and interaction between these conditions. A difference was observed among the free field positions tested, except in the situations of noise right and noise left. Results of speech perception in adults with normal hearing in the free field during different listening situations in noise indicated poorer performance during the condition with noise and speech in front, i.e., 0°/0°. The values found in the standardization of the Hearing in Noise Test free field can be used as a reference in the development of protocols for tests of speech perception in noise, and for monitoring individuals with hearing impairment.


Involvement of chromosomes 7 and 12 in large bowel cancer: trisomy 7 and 12q-.
Cytogenetic analysis of an adenocarcinoma of the large bowel with some villous features revealed consistent chromosome changes: trisomy of chromosomes No. 7, 12, and 20 and a deletion of part of the long arm of a chromosome No. 12. Examination of the chromosome findings in previously published cases of large bowel cancers and the data presented in this paper lead us to suggest that trisomy No. 7 and 12q- may be nonrandom (primary) karyotypic changes in a subgroup of large bowel cancers.


Statutory provisions of the Health and Safety at Work Act 1974.
Case Scenario: Rose is working as a staff nurse in the operating theatre of Roger Park Hospital when there is an explosion. She is severely burnt. Following an inspection by the Health and Safety Inspectorate, the Roger Park NHS Trust is informed that it will be prosecuted. It appears that the cause of the explosions was the result of gases igniting because of an electrical fault in a patient monitor.


A magic kick for regeneration: role of mesenchymal stromal cell secretome in spermatogonial stem cell niche recovery.
Injury of stem cell niches may disturb tissue homeostasis and regeneration coordinated by specific niche components. Yet, the mechanisms of stem cell niche restoration remain poorly understood. Herein, we examined the role of mesenchymal stromal cells (MSCs) as pivotal regulators of stem cell niche recovery focusing on the effects of their secretome. The spermatogonial stem cell (SSC) niche was selected as a model. SSC niches were injured by inducing abdominal cryptorchidism in rats. Briefly, testes of anesthetized rats were elevated into the abdominal cavity through the inguinal canal for 14 days. After descent of testes, MSC or MSC secretome treatment was applied to the animals by local subtunical injections. Local administration of MSC or MSC secretome was sufficient to recover spermatogenesis and production of functional germ cells. The effects of MSC and their secreted components were comparable, leading to restoration of Sertoli cell pools and recovery of Leydig cell secretory functions. Our data suggest that MSCs mimic the functions of lost supportive cells within the stem cell niche, transiently providing paracrine stimuli for target cells and triggering tissue regenerative processes after damage.