TM-Aligner: Multiple sequence alignment tool for transmembrane proteins with reduced time and improved accuracy.
Membrane proteins plays significant role in living cells. Transmembrane proteins are estimated to constitute approximately 30% of proteins at genomic scale. It has been a difficult task to develop specific alignment tools for transmembrane proteins due to limited number of experimentally validated protein structures. Alignment tools based on homology modeling provide fairly good result by recapitulating 70-80% residues in reference alignment provided all input sequences should have known template structures. However, homology modeling tools took substantial amount of time, thus aligning large numbers of sequences becomes computationally demanding. Here we present TM-Aligner, a new tool for transmembrane protein sequence alignment. TM-Aligner is based on Wu-Manber and dynamic string matching algorithm which has significantly improved its accuracy and speed of multiple sequence alignment. We compared TM-Aligner with prevailing other popular tools and performed benchmarking using three separate reference sets, BaliBASE3.0 reference set7 of alpha-helical transmembrane proteins, structure based alignment of transmembrane proteins from Pfam database and structure alignment from GPCRDB. Benchmarking against reference datasets indicated that TM-Aligner is more advanced method having least turnaround time with significant improvements over the most accurate methods such as PROMALS, MAFFT, TM-Coffee, Kalign, ClustalW, Muscle and PRALINE. TM-Aligner is freely available through http://lms.snu.edu.in/TM-Aligner/ .


Spatial variation in the degradation rate of the pesticides isoproturon, azoxystrobin and diflufenican in soil and its relationship with chemical and microbial properties.
The extent of within field variability in the degradation rate of the pesticides isoproturon, azoxystrobin and diflufenican, and the role of intrinsic soil factors and technical errors in contributing to the variability, was investigated in sites on sandy-loam and clay-loam. At each site, 40 topsoil samples were taken from a 160 x 60 m area, and pesticides applied in the laboratory. Time to 25% dissipation (DT25) ranged between 13 and 61 weeks for diflufenican, 5.6 and 17.2 weeks for azoxystrobin, and 0.3 and 12.5 weeks for isoproturon. Variability in DT25 was higher in the sandy-loam in which there was also greatest variability in soil chemical and microbial properties. Technical error associated with pesticide extraction, analysis and lack of model fit during derivation of DT25 accounted for between 5.3 and 25.8% of the variability for isoproturon and azoxystrobin, but could account for almost all the variability for diflufenican. Azoxystrobin DT25, sorption and pH were significantly correlated.


Validation of the Prague C & M criteria for the endoscopic grading of Barrett's esophagus by gastroenterology trainees: a multicenter study.
The Prague C & M criteria, developed for the endoscopic grading of Barrett's esophagus (BE), (C = circumferential length, M = maximal length) were previously validated among a panel of 29 expert endoscopists with a special interest in BE. Its performance among gastroenterology trainees is unknown. To test interobserver agreement among gastroenterology trainees for the Prague C & M criteria, identification of the gastroesophageal junction (GEJ) and the diaphragmatic hiatus. A prospective study. Two tertiary referral centers. Standardized endoscopic videos were used. Interobserver agreement. Eighteen high-quality videos (normal esophagus, short and long lengths of BE, equally distributed) were independently evaluated by 18 gastroenterology trainees (year 1, n = 5; year 2, n = 6; year 3, n = 7) after administration of a formal teaching module by an expert endoscopist. Overall intraclass correlation coefficients for assessment of the C and M extent of the endoscopic BE segment above the GEJ were 0.94 (95% CI, 0.89-0.98) and 0.96 (95% CI, 0.94-0.98), respectively. The overall intraclass correlation coefficients for GEJ and diaphragmatic hiatus location recognition were 0.92 (0.86-0.96) and 0.90 (0.82-0.95), respectively. The year of training did not affect interobserver agreement. The use of videos for endoscopic evaluation. After standardized teaching, the Prague C & M criteria have high overall validity among gastroenterology trainees irrespective of the level of training for endoscopic evaluation of visualized BE lengths as well as key endoscopic landmarks.


Ultrasmall dual-modality silica nanoparticle drug conjugates: Design, synthesis, and characterization.
The physicochemical design and synthesis of effective cancer-directed and particle-based nanotherapeutic imaging agents remains a challenging task. Of critical importance is the ability to demonstrate maximum delivery, retention, and treatment efficacy for platforms designed to deposit their cargo at sites of disease without attendant dose-limiting toxicity. In this work, we describe dual-modality nanoparticle drug conjugates (NDCs) which utilize protease sensitive linkers to attached drug compounds and imaging labels to a clinically translated class of ultrasmall silica nanoparticle (C' dots). We describe the synthesis and characterization of these linker-drug constructs. Linkers incorporating dipeptide enzyme substrates are attached to analogs of a prototypical epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), through a cleavable amide bond or para-aminobenzyloxycarbonyl (PABC) group. These constructs are conjugated onto C' dots leading to the desired NDCs. These NDCs exhibit fast and predictable release kinetics in the presence of model proteases, and are stable in various biological media. Finally, in vitro assays show NDCs to be highly active in reducing phosphorylated EGFR levels in H1650 cells, a human tumor-derived cell line. The data suggests that NDCs exhibit desirable properties that warrant further development toward oncological therapy.


Quantitative texture analysis in two-dimensional echocardiography: application to the diagnosis of myocardial amyloidosis.
Qualitative and subjective analysis of two-dimensional echocardiographic images of the myocardial wall allows one to identify amyloid heart disease; the quantitative analysis of regional image texture might be an accurate method to differentiate normal from amyloid myocardial structures. To test this hypothesis, two-dimensional echocardiograms of nine normal subjects and six patients with histologically documented amyloid heart disease were evaluated. Quantitative texture measurements of the first order (mean gray level, skewness, kurtosis, energy and entropy) overlapped between the two groups. Among the second order statistics variables, entropy was significantly and consistently higher in amyloid versus normal patient data (septum in parasternal long-axis view: 6.3 +/- 0.3 versus 5.9 +/- 0.4; septum in apical four chamber view: 6.2 +/- 0.2 versus 5.8 +/- 0.3). Therefore, amyloid-involved myocardial walls show ultrasound image texture alterations that may be quantified with digital image analysis techniques.


Effects of cyclosporine in severe systemic lupus erythematosus.
Thirteen patients with severe steroid-resistant or steroid-dependent forms of systemic lupus erythematosus were treated with cyclosporine (average dose 5 mg/kg/d) for an average period of 12 months. In eight patients the disease activity decreased, as substantiated by the reduction in the amount of steroid required to control the clinical manifestations. Interruption of cyclosporine treatment was associated with relapse or worsening of disease in five subjects. These favorable clinical results occurred in the absence of changes in the levels of antinuclear, anti-double-stranded deoxyribonucleic acid autoantibodies or plasma complement components; plasma IgG concentration increased significantly. Six patients had signs of moderate cyclosporine nephrotoxicity that disappeared when the administration of the drug was discontinued. Hypertension was the most serious side effect observed in eight subjects; in every case it was controlled by antihypertensive medicine. These data indicate that cyclosporine may be beneficial in the treatment of some patients with severe forms of systemic lupus erythematosus.


GYT: Get Yourself Tested Campaign Awareness: Associations With Sexually Transmitted Disease/HIV Testing and Communication Behaviors Among Youth.
The GYT: Get Yourself Tested campaign promotes sexually transmitted disease (STD) and HIV testing and communication with partners and providers among youth. We evaluated these behaviors in relation to campaign awareness among youth through a national survey. We collected data from 4017 respondents aged 15 to 25 years through an online panel survey designed to be representative of the US population. The GYT campaign targeted 4 key behaviors: STD testing, HIV testing, talking to partners about testing, and talking to providers about testing. Respondents who were aware of the GYT campaign (24.4%) were more likely to report engaging in each of the 4 target behaviors. Associations remained significant when stratified by race and sex and when taking into account sexuality, sexual activity, age, insurance status, and use of campaign partner-provided services. Awareness of the GYT campaign is related to the 4 target behaviors promoted by the campaign, suggesting that health promotions campaigns oriented toward youth can be successful in increasing STD-related, health-seeking behavior, including among populations disproportionately affected by STD.


Reliability of age estimation from iliac auricular surface in a subactual Chilean sample.
The aim of this study was to test the reliability of Osborne et al. (2004) [8] age estimation based on the iliac auricular surface of the ilium. We selected 172 skeletons with documented sex and age-at-death and established six uniformly distributed age intervals for analysis. ANOVA was employed to assess the association of the documented age with sex and the auricular surface classification. We employed Bonferroni post-hoc tests to find any statistical differences across documented ages within each phase of Osborne's criteria. While the ANOVA showed a significant association between the documented age and the auricular surface classification, post-hoc tests found that phases 2 and 3 were the only consecutive phases with significant differences. We argue that a lack of statistical significance between consecutive phases undermines the reliability of this method for forensic purposes especially in middle adults. It may be possible that the collapse of intermediate phases would increase the accuracy of this method.


Protein damage, radiation sensitivity and aging.
This paper promotes a concept that protein damage determines radiation resistance and underlies aging and age-related diseases. The first bottleneck in cell recovery from radiation damage is functional (proteome) rather than informational (DNA), since prokaryotic and eukaryotic cell death correlates with incurred protein, but not DNA, damage. Proteome protection against oxidative damage determines survival after ionizing or UV irradiation, since sufficient residual proteome activity is required to turn on the DNA damage response activating DNA repair and protein renewal processes. Extreme radiation and desiccation resistance of rare bacterial and animal species is accounted for by exceptional constitutive proteome protection against oxidative damage. After excessive radiation their well-protected proteome faithfully reconstitutes a transcription-competent genome from hundreds of DNA fragments. The observation that oxidative damage targeted selectively to cellular proteins results in aging-like phenotypes suggests that aging and age-related diseases could be phenotypic consequences of proteome damage patterns progressing with age.


Pathologic features associated with fibrosis in nonalcoholic fatty liver disease.
Nonalcoholic fatty liver disease (NAFLD) represents a spectrum of clinicopatholologic conditions ranging from steatosis alone to nonalcoholic steatohepatitis (NASH), with varying risks for progression to cirrhosis. Although steatosis alone seems to be nonprogressive, some patients with NASH can progress. This study focuses on the clinical and pathological characteristics of patients with NAFLD associated with the development of histological fibrosis. Patients with an established diagnosis of nonalcoholic fatty liver were identified through our NAFLD database containing extensive clinical, demographic, and laboratory data. Liver biopsy specimens were read blindly by one hepatopathologist using a 19-item pathological protocol and by another hepatopathologist using a second pathological protocol. Clinical and pathological data were matched to the presence of different types of histological fibrosis. Univariate and multivariate analyses helped determine all of the variables independently associated with histological fibrosis. Of 132 NAFLD patients, 21.2% had advanced fibrosis (septal/bridging fibrosis or well-established cirrhosis). Sinusoidal fibrosis was present in 20.3% of patients, whereas perivenular fibrosis was seen in 17.2%. Ballooning degeneration and Mallory bodies were independently associated with both sinusoidal fibrosis and perivenular fibrosis. Aspartate aminotransferase/alanine aminotransferase ratio and ballooning degeneration were also independently associated with periportal-portal fibrosis. We conclude that the presence of hepatocyte injury in NAFLD is associated with fibrosis. These pathological features can be used to establish the pathological criteria for diagnosis of the progressive form of NAFLD or NASH.


Visualization and molecular analysis of nuclear import of protein kinase CK2 subunits in living cells.
We have generated fusion proteins between the subunits of CK2 and GFP and characterized their behaviour in living cells. The expressed fusion proteins were functional and interacted with endogenous CK2. Imaging of NIH3T3 cells expressing low level of GFP-CK2alpha or GFP-CK2beta showed that both proteins were mostly nuclear in interphase. Both CK2 subunits contain nuclear localization domains that target them independently to the nucleus. Once in the nucleus, both subunits diffused rapidly in the nucleoplasm. In mitotic cells, CK2 subunits were dispersed throughout the cytoplasm and were not associated to chromatin. Our data are compatible with the idea that each subunit can translocate individually to the nucleus to interact with each other or with important cellular partners. Understanding the molecular mechanisms which regulate the dynamic localization of CK2 subunits will be of central importance.


Incidence of Major Arterial Abnormality in Patients with Wound Dehiscence after Lower Extremity Orthopedic Procedures.
Surgical dehiscence can occur after lower extremity orthopedic procedures. Underlying vascular aberrancy and localized ischemia contribute to chronic wound development requiring advanced techniques such as free tissue transfer. Localized vascular abnormality is an underrecognized contributing factor to such dehiscence. The authors reviewed their lower extremity free tissue transfer experience in this population to analyze the incidence of arterial abnormality and outcomes. The authors conducted a retrospective review of 64 lower extremity free tissue transfers performed for chronic wounds after orthopedic procedures from 2011 to 2018. The primary outcome was major arterial abnormality as identified on angiography. Secondary outcomes were flap success, limb salvage, and ambulation status. The median age was 58 years, and 44 were men (69 percent). Comorbidities included osteomyelitis (77 percent), diabetes (39 percent), and peripheral vascular disease (17 percent). The incidence of arterial abnormality on angiography was 47 percent. Defect location correlated with angiosome of arterial abnormality in 53 percent. The flap success rate was 92 percent. Limb salvage and ambulation rates were 89 and 89 percent, respectively, at an average follow-up of 17.6 months. Men demonstrated an increased rate of limb salvage (p = 0.026). Diabetes (p = 0.012), arterial abnormality (p = 0.044), and arterial flap complication (p = 0.010) correlated with amputation. The high incidence of arterial abnormality in this population highlights the importance of expedient multidisciplinary care, including vascular and plastic surgery. Diagnostic angiography is important for identifying major arterial abnormality and the need for free tissue transfer for definitive coverage.


Growth and Nutritional Status of Phenylketonuric Children and Adolescents.
The goal of this study was to assess the anthropometric and biochemical parameters of children and adolescents with phenylketonuria (PKU). The participants in this cross-sectional study ranged in age from four to 18 years old. Biochemical markers such as vitamin B12, folic acid, iron, ferritin, calcium, 25-hydroxy vitamin D3, zinc, plasma phenylalanine (Phe) and tyrosine (Tyr) levels in blood were evaluated, as well as demographics and anthropometric measurements. A three-day dietary recall questionnaire was completed by all individuals. 80% (64) of the 80 patients (42 females, 52.5%) had typical PKU. Consanguineous marriages were found in 57.5% (46) of the patients' parents. According to the height for age index, 17.5% of the study group (n = 14) were short or very short. According to age-related weight and body mass index (BMI), 37.5% (n = 30) and 43.8% (n = 35) of people are obese or overweight, respectively. Biochemical tests revealed increased vitamin B12 levels and 25-hydroxy vitamin D3 deficiency in 35% (n = 28) of the patients, insufficient folic acid in 12.5% (n = 10), and elevated phenylalanine levels in 70.3% (n = 45) of children under 12 years old, and adolescents 62.5% (n = 10). A high Phe intake (OR = 4.44, CI %95 = 1.27-15.57) is a risk factor for obesity and overweight. Patients with PKU had a high rate of overweight and obesity. PKU patients who are overweight or obese do not differ from normal-weight patients in terms of dietary intake or laboratory findings (except for serum iron levels). One-third of patients with phenylketonuria were vitamin D deficient and had a BMI/A index of overweight/obese. It is recommended to use special medical food to help solve energy and nutrient deficiencies.


In vitro and in vivo studies on potentiation of cytotoxic effects of anticancer drugs or cobalt 60 gamma ray by interferon on human neoplastic cells.
A possibility that interferon may potentiate the cytotoxic effects of anticancer drugs or 60Co gamma ray on human neoplastic cells was studied by in vitro and in vivo experimental procedures. The human neoplastic cells used were HeLa (uterine cervical cancer) and WI-38 CT-1 (embryonic lung fibroblasts transformed in culture by 60Co gamma ray) cells. As normal human cells, WI-38 cells were used. Interferon was a preparation of beta-type produced by human fibroblasts. The cytotoxicity was determined by colony formation for in vitro experiments and by tumor growth for animal experiments. Of 17 anticancer drugs, the cytotoxic effects of six drugs, namely, peplomycin, bleomycin, aclacinomycin, cisplatin, 5-fluorouracil (5-FU), and Adriamycin (doxorubicin) were potentiated by concomitant application of interferon. The cytolethal effects of 60Co gamma ray were also enhanced by interferon. The growth of tumor induced by transplantation of HeLa cells into a nude mouse was remarkably reduced by combination therapy of interferon and 5-FU. The current results indicate a possibility that combined therapy of certain types of anticancer drugs or 60Co gamma ray with interferon may be effective in treatment of cancer patients.


The neurophenomenology of early psychosis: An integrative empirical study.
The integration of various domains or levels of analysis (clinical, neurobiological, genetic, etc.) has been a challenge in schizophrenia research. A promising approach is to use the core phenomenological features of the disorder as an organising principle for other levels of analysis. Minimal self-disturbance (fragility in implicit first-person perspective, presence and agency) is emerging as a strong candidate to play this role. This approach was adopted in a previously described theoretical neurophenomenological model that proposed that source monitoring deficits and aberrant salience may be neurocognitive/neurobiological processes that correlate with minimal self-disturbance on the phenomenological level, together playing an aetiological role in the onset of schizophrenia spectrum disorders. The current paper presents full cross-sectional data from the first empirical test of this model. Fifty ultra-high risk for psychosis patients, 39 first episode psychosis patients and 34 healthy controls were assessed with a variety of clinical measures, including the Examination of Anomalous Self-Experience (EASE), and neurocognitive and neurophysiological (EEG) measures of source monitoring deficits and aberrant salience. Linear regression indicated that source monitoring (composite score across neurocognitive and neurophysiological measures), with study group as an interaction term, explained 39.8% of the variance in EASE scores (R2 = 0.41, F(3,85) = 14.78, p < 0.001), whereas aberrant salience (composite score) explained only 6% of the variance in EASE scores (R2 = 0.06, F(3,85) = 1.44, p = 0.93). Aberrant salience measures were more strongly related to general psychopathology measures, particularly to positive psychotic symptoms, than to EASE scores. A neurophenomenological model of minimal self-disturbance in schizophrenia spectrum disorders may need to be expanded from source monitoring deficits to encompass other relevant constructs such as temporal processing, intermodal/multisensory integration, and hierarchical predictive processing. The cross-sectional data reported here will be expanded with longitudinal analysis in subsequent reports. These data and other related recent research show an emerging picture of neuro-features of core phenomenological aspects of schizophrenia spectrum disorders beyond surface-level psychotic symptoms.


Actions of endothelin and corticotropin releasing factor in the guinea-pig ileum: no evidence for an interaction with capsaicin-sensitive neurons.
Both endothelins and corticotropin releasing factor (CRF) appear in capsaicin-sensitive neurons. We have investigated the effects of human endothelin-1 (ET-1) and CRF in the guinea-pig ileum longitudinal and circular preparations and sought for ways of specific antagonism. With the aid of tachyphylaxis to capsaicin (i.e., rendering capsaicin-sensitive neurons functionally impaired) it was tested if these neurons played a mediating role in the effects of ET-1 or CRF. We also tried to find out whether endogenous endothelin or CRF plays a role in the excitatory and inhibitory effects of capsaicin in the ileum. In preparations at basal tone, both exogenous ET-1 (1-100 nM) and CRF (3-100 nM) caused contraction. These responses were not influenced by capsaicin tachyphylaxis. The contractile effect of ET-1 was not affected by tetrodotoxin (1 microM), atropine (1 microM), methysergide (100 nM), chloropyramine (100 nM) or SR140333 (100 nM) but was significantly inhibited or even abolished by the receptor antagonist BQ123 (3 microM) or BQ788 (3 microM). CRF caused contraction that was fully sensitive to tetrodotoxin (1 microM), tachyphylaxis to CRF or to atropine (1 microM) plus the tachykinin NK1 receptor antagonist SR140333 (200 nM). Atropine alone had a weak inhibitory effect on the contractile action of CRF. Neither the antagonist BQ123 (3 microM) nor CRF tachyphylaxis inhibited the contractile action of capsaicin (2 microM), even in the presence of a mixture of GR82334 (3 microM) and SR142801 (100 nM), for blocking tachykinin NK1 and NK3 receptors, respectively--a treatment that by itself significantly reduced the effect of capsaicin. Exogenous ET-1 (0.3-5 nM), but not CRF (30-100 nM), caused relaxation of the atropine-treated, histamine-precontracted ileum. This effect of ET-1 was significantly inhibited or abolished by BQ123 (10 microM), or BQ788 (3 microM), but was not influenced by capsaicin tachyphylaxis. Likewise, relaxation of the atropine-treated, histamine-precontracted ileum in response to capsaicin was not influenced by the endothelin receptor antagonist BQ788 (3 microM) or BQ788 (3 microM) plus BQ123 (3 microM). Apamin (300 nM) was also without effect on the capsaicin-induced relaxation. In circular muscle strips ET-1 inhibited the indomethacin-induced spontaneous activity. This effect was abolished by BQ123 (3 microM) or BQ788 (3 microM). CRF caused a stimulation of the circular muscle. This stimulatory effect was not influenced by atropine (1 microM) alone, but was inhibited by atropine plus tachykinin NK1 and NK2 receptor antagonists (SR140333 (200 nM) and SR48968 (200 nM)) and also by tetrodotoxin (1 microM). It is concluded that capsaicin-sensitive neurons do not play a role in the effects of exogenous ET-1 or CRF in the guinea-pig ileum. ET-1 can both contract and relax the ileal longitudinal smooth muscle directly, probably via both ETA and ETB receptors. CRF acts by specifically stimulating excitatory (but not inhibitory) neurons of the myenteric plexus. Neither endogenous ET-1 nor CRF seems to play a role in the excitatory or inhibitory effects of capsaicin.


Hepatic expression of cytochrome P450 in type 2 diabetic Goto-Kakizaki rats.
Although hepatic expression of cytochrome P450 (CYP) changes markedly in diabetes, the role of ketone bodies in the regulation of CYP in diabetes is controversial. The present study was performed to determine the expression and activity of CYP in non-obese type II diabetic Goto-Kakizaki (GK) rats with normal levels of ketone bodies. In the present study, basal serum glucose levels increased 1.95-fold in GK rats, but acetoacetate and β-hydroxybutyrate levels were not significantly different. Hepatic expression of CYP reductase and CYP3A2 was up-regulated in the GK rats, and consequently, activities of CYP reductase and midazolam 4-hydroxylase, mainly catalyzed by CYP3A2, increased. In contrast, hepatic expression of CYP1A2 and CYP3A1 was down-regulated and the activities of 7-ethoxyresorufin-O-deethylase and 7-methoxyresorufin-O-demethylase, mainly catalyzed by CYP1A, also decreased in GK rats. Hepatic levels of microsomal protein and total CYP and hepatic expression of cytochrome b(5), CYP1B1, CYP2B1 and CYP2C11 were not significantly different between the GK rats and normal Wistar rats. Moreover, the expression and activity of CYP2E1, reported to be up-regulated in diabetes with hyperketonemia, were not significantly different between GK rats and control rats, suggesting that elevation of ketone bodies plays a critical role in the up-regulation of hepatic CYP2E1 in diabetic rats. Our results showed that the expression of hepatic CYP is regulated in an isoform-specific manner. The present results also show that the GK rat is a useful animal model for the pathophysiological study of non-obese type II diabetes with normal ketone body levels.


Undergraduate nursing students' experience of learning respiratory system assessment using flipped classroom: A mixed methods study.
Knowledge and skill acquisition to perform an accurate respiratory system assessment is a key competency expected in undergraduate nursing students. Learning physical assessment requires the integration of multiple knowledge bases and skills; hence, applying an innovative teaching approach, such as the flipped-classroom (FC) approach, fosters an active and student-centered learning environment for physical assessment class. This study evaluated FC's feasibility in delivering respiratory system assessment content in a health assessment course and explored the changes in nursing students' perceptions regarding student-centeredness and active learning environments before and after applying FC. A single group pre- and post-test concurrent mixed-methods design was used. This study was conducted in a private nursing college in South Korea. A convenience sample of 91 second year undergraduate nursing students enrolled in a health assessment course. FC was offered at one didactic session of a physical assessment course. In the FC, students completed a self-directed pre-class activities using online lecture videos and reading materials prior to the class and participated in interactive team-based learning activities inside the classroom. Skills lab practicum took place after the FC. Students' perceptions regarding student-centeredness and active learning environments, in terms of teaching, social, and cognitive presences were measured before (T1) and after (T2) conducting the FC. Qualitative data were obtained at T2 using free-response questions, which required students to comment on their FC experience. Participants' perceptions of student-centeredness significantly increased from T1 to T2. Although student-perceived teaching and social presence in their learning environment showed upward trends from T1 to T2, these changes were not statistically significant. Students considered FC an acceptable approach to foster active learning in a supportive learning environment. This study revealed that incorporating FC to deliver respiratory system assessment content was feasible and considered acceptable by undergraduate nursing students.


Progressive myoclonic epilepsy type 1 (EPM1) patients present with abnormal 1H MRS brain metabolic profiles associated with cognitive function.
Progressive myoclonic epilepsy, type 1A (EPM1, Unverricht-Lundborg disease), is a rare neurodegenerative autosomal recessive disorder characterized by stimulus-sensitive and action myoclonus and tonic-clonic epileptic seizures. Patients develop neurological symptoms, including ataxia, intention tremor, and dysarthria, over time, with relatively limited and nonspecific MRI atrophy findings. The effects of the disease on brain metabolism are largely unknown. Eighteen EPM1 patients (9 M, 9F) underwent clinical evaluation and neuropsychological testing, which included the assessment of intellectual ability, verbal memory, and psychomotor and executive functions. Magnetic resonance spectroscopy (MRS) and imaging (MRI) were performed on a 1.5 T MRI system. 2D MRS chemical shift imaging (CSI) maps (TE = 270) were obtained from the following regions of the brain: basal ganglia, thalamus, insula, splenium, and occipital white and gray matter, and N-acetyl-aspartate (NAA)-, choline (Cho)-, and lactate (Lac)-to-creatine (Cr) ratios were analyzed. Ten healthy age-and sex-matched subjects (5M, 5F) were used as controls for MRS. We found significant brain metabolic changes involving lactate, NAA, and choline, which are widespread in the basal ganglia, thalamic nuclei, insula, and occipital areas of EPM1 patients. Changes, especially in the right insula, basal ganglia, and thalamus, were associated with intellectual abilities and impairment of the psychomotor and executive functions of EPM1 patients. Multiple brain metabolic alterations suggest the presence of neurodegeneration associated with EPM1 progression. The changes in metabolite ratios are associated with the neurocognitive dysfunction caused by the disease. However, the role of MRS findings in understanding pathophysiology of EPM1 warrants further studies.


Combined prognostic significance of white blood cell count and D-dimer on in-hospital outcomes of acute ischemic stroke.
The prognostic significance of combination of white blood cell (WBC) and D-dimer on acute ischemic stroke (AIS) remains to be explored. We aimed to investigate the combined effect of WBC and D-dimer levels on in-hospital outcomes of AIS patients. 801 AIS patients were included. Patients were divided into four groups according to the cut-point identified by receiver operating characteristic (ROC) curve of D-dimer (1.105 μg/L) and WBC (7.05 × 109/L): LWLD (low WBC count and low D-dimer), LWHD (low WBC count and high D-dimer), HWLD (high WBC count and low D-dimer), and HWHD (high WBC count and high D-dimer). HWHD group had the highest cumulative incidence of in-hospital mortality (hazard ratio, 5.79; 95%CI, 1.71-19.58, P = 0.006). Patients in HWHD group were 4.14 fold more likely to have in-hospital pneumonia (odds ratio, 4.14; 95%CI, 2.09-8.21; P < 0.001), compared with those in LWLD group. The area under curve (AUC) of the combination of WBC and D-dimer levels for in-hospital mortality and pneumonia was larger than that of WBC and D-dimer alone (0.920 vs. 0.900 vs. 0.915; 0.831 vs. 0.829 vs. 0.807). The combination of WBC count and D-dimer levels at admission was independently associated with in-hospital outcomes of AIS patients. The addition of WBC to D-dimer levels had a tendency to improve the predictive power for in-hospital mortality and pneumonia.


Cognition, criminal conduct, and virtual reality: Understanding and reducing offending using simulated environments.
Over the past years, research has shown that virtual reality (VR) technology can be used to observe, interpret, and change human behavior and cognition in a variety of domains. This chapter explores the potential of VR as a tool to observe, interpret, and change human behavior and cognition as they relate to antisocial behavior. We review the criminological research literature as well as research literature from related disciplines on VR applications that has focused on observing and reducing antisocial behavior. The main findings of our review suggest that the key merits of VR in the domain of crime and antisocial behavior are its ability to provide safe learning environments that would otherwise involve risk, the possibility of generating ethical and ecologically valid virtual alternatives for real-life situations, and the development of stimuli that are impossible to create in real life. These unique characteristics make VR a promising tool to observe criminal behavior as it takes place and develop intervention programs to reduce antisocial behavior.


Development and application of a monoclonal rat peroxidase antiperoxidase (PAP) immunocytochemical reagent.
Monoclonal antibodies are being increasingly used in immunocytochemistry but their localisation by the peroxidase antiperoxidase (PAP) procedure requires the use of rat or mouse PAP. In this paper we describe the development and application of a monoclonal rat PAP. This reagent has been used successfully for immunocytochemistry at light and electron microscopy level in combination with rat monoclonal antibodies against serotonin (5-HT), substance P and somatostatin. The monoclonal rat PAP has several advantages over conventional polyclonal rat PAP and is likely to be a valuable developing reagent in immunocytochemistry using rat monoclonal antibodies.


Endogenous calcitonin gene-related peptide in cerebrospinal fluid and early quality of life and mental health after good-grade spontaneous subarachnoid hemorrhage-a feasibility series.
The vasodilatory calcitonin gene-related peptide (CGRP) is excessively released after spontaneous subarachnoid hemorrhage (sSAH) and modulates psycho-behavioral function. In this pilot study, we prospectively analyzed the treatment-specific differences in the secretion of endogenous CGRP into cerebrospinal fluid (CSF) during the acute stage after good-grade sSAH and its impact on self-reported health-related quality of life (hrQoL). Twenty-six consecutive patients (f:m = 13:8; mean age 50.6 years) with good-grade sSAH were enrolled (drop out 19% (n = 5)): 35% (n = 9) underwent endovascular aneurysm occlusion, 23% (n = 6) microsurgery, and 23% (n = 6) of the patients with perimesencephalic SAH received standardized intensive medical care. An external ventricular drain was inserted within 72 h after the onset of bleeding. CSF was drawn daily from day 1-10. CGRP levels were determined via competitive enzyme immunoassay and calculated as "area under the curve" (AUC). All patients underwent a hrQoL self-report assessment (36-Item Short Form Health Survey (SF-36), ICD-10-Symptom-Rating questionnaire (ISR)) after the onset of sSAH (t1: day 11-35) and at the 6-month follow-up (t2). AUC CGRP (total mean ± SD, 5.7 ± 1.8 ng/ml/24 h) was excessively released into CSF after sSAH. AUC CGRP levels did not differ significantly when dichotomizing the aSAH (5.63 ± 1.77) and pSAH group (5.68 ± 2.08). aSAH patients revealed a higher symptom burden in the ISR supplementary item score (p = 0.021). Multiple logistic regression analyses corroborated increased mean levels of AUC CGRP in CSF at t1 as an independent prognostic factor for a significantly higher symptom burden in most ISR scores (compulsive-obsessive syndrome (OR 5.741, p = 0.018), anxiety (OR 7.748, p = 0.021), depression (OR 2.740, p = 0.005), the supplementary items (OR 2.392, p = 0.004)) and for a poorer performance in the SF-36 physical component summary score (OR 0.177, p = 0.001). In contrast, at t2, CSF AUC CGRP concentrations no longer correlated with hrQoL. To the best of our knowledge, this study is the first to correlate the levels of endogenous CSF CGRP with hrQoL outcome in good-grade sSAH patients. Excessive CGRP release into CSF may have a negative short-term impact on hrQoL and emotional health like anxiety and depression. While subacutely after sSAH, higher CSF levels of the vasodilator CGRP are supposed to be protective against vasospasm-associated cerebral ischemia, from a psychopathological point of view, our results suggest an involvement of CSF CGRP in the dysregulation of higher integrated behavior.


[Impact of the suicidal crisis intervention training program on the confidence and skills of hospital professionals in the Hauts-de-France region].
Suicide is a major public health issue given its huge human and economic consequences. Symptoms prior to suicide are often not specific. Nevertheless, the majority of suicidal people express suicidal thoughts, and nearly one in two meet a health professional in the period preceding the act. Being able to recognize the warnings and intervene during the suicidal crisis, defined as a mental crisis where the major risk is suicide, is to seize the opportunity to postpone the suicidal plan and to gain time to implement in place lasting strategies to combat suffering. Thus, the training for suicidal crisis intervention is a major axis of the suicide prevention strategy. Recently, crisis intervention training programs have been updated with knowledge accumulated since the early 2000's. In France, one of the countries most concerned by suicide, the Hauts-de-France region is one of the most impacted. In this context, the Regional Health Agency of Hauts-de-France included in its Regional Health Program of 2018-2023 the training of healthcare workers who work with high suicidal risk patients. The suicidal crisis intervention training program (SCIT) has been introduced to hospital staffs in Hauts-de-France. The purpose of this study was to evaluate this program. Eight training sessions with 15 to 21 participants were carried out from 2019 November to 2021 January in the Hauts-de-France region. Participants were volunteer healthcare professionals in direct contact with suicidal crisis patients. The training included three modules. The first one concerned the suicidal crisis intervention training: definition of the suicidal crisis, typology of the crisis, vulnerability development, crisis evaluation and crisis intervention practice. The second concerned the evaluation with the RED scale (Risk-Emergency-Danger) and the adequate patient orientation to a psychiatric unit. The third was dedicated to the Gatekeeper training with the constitution of a Gatekeeper network to enhance the capacity to detect suicidal risk and to orient the concerned person towards an adequate evaluation or care organization. We evaluated the first two levels of the Kirkpatrick's model: level 1) the participant's satisfaction (rated out of 10), and level 2) the degree of confidence in their professional abilities (rated out of 10) and their skills in responding to a person in a suicidal crisis (using the SIRI-2-VF - French version of the Suicide Intervention Response Inventory-2). The participants were interviewed before (T0), just after (T1) and at one month of training (T2). Among the 141 health professionals who followed the training, 139 answered the questionnaire at least one time (13 psychologists, 22 doctors, 97 nurses and 7 head nurses). The participation rates were 99.3 % at T0, 96.4 % at T1 and 46.0 % at T2. Most of the participants were nurses (69.8 %), and 33.1 % of the respondents declared they had already followed a suicidal crisis training. The satisfaction with the training was evaluated at 8.6 (± 1.3) out of 10. There was no significant difference among the professions, neither between those having already received or not a previous training. The self-perceived capacity to manage a suicidal crisis was rate 6.8 (± 1.8) out of 10 at T0. There was a significant increase just after the training (8.1±1.2 vs 6,8±1,8, p<0,001) which persisted at 1 month (8.1±1.1 vs 6.8±1.8, P<0.001). The score at the SIRI-2-VF was 15.0 (± 4.2) out of 30 at T0. There was a significant increase just after the training (17.5±3.5 vs 15.0±4.2, P<0.001), which persisted at 1 month (17.0±4.0 vs 15.0±4.2, P<0.001). This is the first evaluation of the suicidal crisis intervention training program. This program increased and homogenized the competency of the participants to manage suicidal ideation and behaviors. Those who followed a previous training maintained higher scores than the others, which shows the importance of repeated training to maintain a satisfying level of knowledge over the long term. One of the strengths of this training is the use of roleplay which enhances the learning and abilities to interact with people at suicidal risk. It seems important to integrate a suicidal crisis intervention training in the cursus of health students to avoid suicide and the dramatic consequences for the entourage and the health professionals who are confronted with it. The SCIT program showed encouraging results in terms of confidence and capacity of the healthcare professionals to intervene in suicidal crisis.


The growth of healthy Australian infants in relation to infant feeding and social group.
A joint survey of infant-feeding practices and infant growth was carried out in Western Australia and Tasmania in 1984-1985. Birthweights and growth from birth to one year of age were similar in both States and to the international growth references. Infants who were never breast-fed or who were breast-fed for only a short time gained significantly more weight over the year (most of it after the age of three months) than did infants who were breast-fed for six months or longer. Infants from families of a lower socioeconomic group gained more weight after six months of age than did those infants from higher socioeconomic families. Analysis showed that the growth differences between the social groups was a result of differences in feeding practices. Linear growth was similar among infants of all social groups, irrespective of the feeding method.


Risk of unplanned visits for colorectal cancer outpatients receiving chemotherapy: a case-crossover study.
This study was conducted to evaluate the impact of chemotherapy on the risk of unplanned visit in a cohort of colorectal cancer outpatients. Chief complaints for unplanned visits and risk factors for hospital admission were also analyzed. Clinical data of 229 consecutive colorectal cancer patients who were unexpectedly presented to our acute oncology clinic between 2006 and 2009 were reviewed. A case-crossover statistical analysis was applied to study the association between exposure to chemotherapy (trigger event) and the occurrence of unplanned visit (acute outcome) in three time windows (7, 15, and 21 days from the closest previous chemotherapy treatment). Cox model was used to assess the risk factors for hospitalization. There were 469 unplanned visits registered. Most of the patients had Eastern Cooperative Oncology Group performance status (ECOG PS) 0-1 (80 %) and advanced cancer stage (78 %). The majority of unplanned visits (72 %) occurred within 30 days since last chemotherapy. The most frequent presenting complaints were pain, fatigue, and anorexia. The two time windows associated with higher risk of visit were 15 and 21 days from last treatment, both for early (odds ratio [OR] 3.8, CI 1.4-10.2 and OR 3.8, CI 1.4-10.2) and advanced disease stage (OR 1.71, CI 1-2.9 and OR 3, CI 1.5-5.9). Of the unplanned visits, 10 % resulted in hospital admission. Presenting with multiple symptoms and with deteriorated PS were both predictors for hospitalization. Chemotherapy exposition triggers the need for unplanned visits over the second and third week after treatment. The prompt and effective management of unexpected events may be cost- and time-saving and reduce pressure on oncology services.


Trehalose metabolism is activated upon chilling in grapevine and might participate in Burkholderia phytofirmans induced chilling tolerance.
During the last decade, there has been growing interest in the role of trehalose metabolism in tolerance to abiotic stress in higher plants, especially cold stress. So far, this metabolism has not yet been studied in Vitis vinifera L., despite the economic importance of this crop. The goal of this paper was to investigate the involvement of trehalose metabolism in the response of grapevine to chilling stress, and to compare the response in plants bacterised with Burkholderia phytofirmans strain PsJN, a plant growth-promoting rhizobacterium that confers grapevine chilling tolerance, with mock-inoculated plants. In silico analysis revealed that the V. vinifera L. genome contains genes encoding the enzymes responsible for trehalose synthesis and degradation. Transcript analysis showed that these genes were differentially expressed in various plant organs, and we also characterised their response to chilling. Both trehalose and trehalose 6-phosphate (T6P) were present in grapevine tissues and showed a distinct pattern of accumulation upon chilling. Our results suggest a role for T6P as the main active molecule in the metabolism upon chilling, with a possible link with sucrose metabolism. Furthermore, plants colonised by B. phytofirmans and cultivated at 26°C accumulated T6P and trehalose in stems and leaves at concentrations similar to non-bacterised plants exposed to chilling temperatures for 1 day. Overall, our data suggest that T6P and trehalose accumulate upon chilling stress in grapevine and might participate in the resistance to chilling stress conferred by B. phytofirmans.


A novel β-xylosidase from Anoxybacillus sp. 3M towards an improved agro-industrial residues saccharification.
An intracellular β-xylosidase (AbXyl), from the thermoalkaline Anoxybacillus sp. 3M, was purified and characterized. The homodimeric enzyme (140 kDa) was optimally active at 65 °C and pH 5.5, exhibited half life of 10 h at 60 °C, 78 and 88% residual activity after 24 h, at pH 4.5 and 8.0, respectively. Fe2+, Cu2+, Al3+, Ag+ and Hg2+ inhibited the enzyme; the activity was moderately stimulated by SDS and not influenced by β-mercaptoethanol. In the presence of p-nitrophenyl-β-d-xylopyranoside, AbXyl exhibited Km of 0.19 mM, Kcat of 453.29 s-1, Kcat Km-1 of 2322 s-1 mM and was moderately influenced by xylose (Ki 21.25 mM). The enzyme hydrolyzed xylo-oligomers into xylose and catalyzed transxylosilation reactions also in presence of alcohols as acceptors, producing xylo-oligosaccharides and alkyl-xylosides. Finally AbXyl was applied towards a statistically optimized process of brewery's spent grain bioconversion, highlighting the important role of this biocatalyst in reaching high yields of fermentable sugars.


[The experimental study of the delayed replantation after cryopreservation of mature tooth in dogs].
To study the conditions of healing of mature tooth in dogs after long cryopreservation followed by replantation. The tooth was extracted after filling the root canals and then preserved for one week and three months respectively by Schwartz's method of cryopreservation of periodontal membrane. The specimens were took out eight weeks after replantation. It was demineralized in a 5% solution of formic acid,and then sectioned perpendicular to the long axes of tooth at a thickness of 5microm. The tooth were sectioned in step-serial sections at 500 microm intervals. The sections were stained with hematoxylin-eosin and followed by evaluation according to Andreasen. The percentage of healing of periodontal membrane (PHPM) of the group of cryopreservation for one week was 64.53%. It was lower than the PHPM of the group of immediate replantation (85%), but the difference demonstrated no significance (P>0.05). The PHPM of the group of cryopreservation for three months was 50.3%. It was lower than the PHPM of the group of immediate replantation significantly (P<0.05). The human's periodontal membrane may be cryopreserved longly and heals after the replantation of tooth as the human's tissues of corneas, skin, islets, et al may be done. The alveolar cavities to be newly made affect the healing of periodontal membrane in some degree.


Suturing techniques of surgeons utilizing two different needle-holder grips.
Sixteen surgeons familiar with grip 1 or grip 2 were randomly selected to test the accuracy of their suture method. The accuracy was tested in two ways: suture with and suture without wavering, while searching for an expected exit point after suturing started and by the distance deviated from a point in a 2 cm suture stride. With grip 1, the first and fourth fingers were put in the rings and the second finger was put around the leg of the holder, whereas with grip 2, all the fingers were put around the holder. Without waver, the mean value was 2 +/- 1.5 mm with grip 1 and 1.4 +/- 1.1 mm with grip 2 (p less than 0.01) and with waver, 1.2 +/- 1.1 mm and 1.1 +/- 10.9 mm (p value not significant). Wavering reduced the deviation (p less than 0.01 and p less than 0.05, respectively). In conclusion, the accuracy was not as precise as the surgeon expected and grip 2 was more accurate, although equivalent accuracy was gained by more wavering with grip 1, thus, there was a greater potential for injuring the tissue.


Cerebral arterial flow dynamics during aneurysm haemorrhage.
A case of intraoperative early rupture of an infraclinoid aneurysm is reported with data showing the specific dynamics of cerebral arterial flow during aneurysm haemorrhage. Internal carotid artery flow and arteriial blood pressure were recorded throughout the surgical procedure. The rupture took place during preparations for dural opening at a mean blood pressure of 160 mmHg and at a low intracranial pressure of 5 to 6 mmHg. The dura became instantly tight, and was non-pulsatile within 10 seconds. The parietal epidural pressure rose rapidly to 110 mmHg. There was a brief initial increase in flow as the haemorrhage started, followed by a marked reduction within seconds. End-diastolic blood flow arrest was observed at this stage, demonstrating the self-limiting effect of a subarachnoid haemorrhage. This impaired cerebral perfusion pressure, or transmural pressure, is discussed with regard to the risk of cerebral damage as well as the beneficial effect in the staunching of haemorrhage. Carotid artery flow showed a moderately impaired autoregulation and an increased vascular resistance suggesting a no-reflow phenomenon subsequent to these events.


Dependent pooling: a contrast-enhanced sign of cardiac arrest during CT.
The purpose of this study is to describe the imaging features of cardiac arrest that occur during CT. CT features of cardiac arrest are characterized by a pooling of contrast agent in the dependent parts of the right side of the body, including the venous system and the right lobe of the liver. If medical professionals are aware of these specific imaging features, prompt cardiac resuscitation can be initiated to avoid permanent brain damage and death.


Changes in prescribing for bipolar disorder between 2009 and 2016: national-level data linkage study in Scotland.
People with bipolar disorder typically require long-term pharmacological treatment to prevent episodes of depression or mania. However, evidence-based guidelines are often not followed by prescribers and, in some countries, prescribing of lithium is in decline. Polypharmacy is also common in bipolar disorder.AimsTo employ a data linkage approach to describe and evaluate prescribing patterns in bipolar disorder in Scotland between 2009 and 2016. By linking prescribing data to the electronic Scottish Morbidity Records, we identified a cohort of 23 135 patients with bipolar disorder who were prescribed psychotropic medication between 2009 and 2016. We examined trends in proportions of patients prescribed each of six drug categories. Random effects logistic models examined change in prescribing over years of interest. The most common form of treatment was antidepressant monotherapy (24.96%), with only 5.90% of patients receiving lithium monotherapy. Prescribing of antipsychotics and anti-epileptics increased from 2009 to 2016 (antipsychotics: odds ratio 1.16, 95% CI 1.15-1.18; anti-epileptics: odds ratio 1.34, 95% CI 1.32-1.36), whereas prescribing of lithium decreased (odds ratio 0.83, 95% CI 0.82-0.85). Prescribing of valproate decreased from 2009-2016 in women, but increased in men (women: odds ratio 0.93, 95% CI 0.90-0.97; men: odds ratio 1.11, 95% CI 1.04-1.18). Antidepressant monotherapy was the most common form of treatment for bipolar disorder in Scotland and prescribing of lithium has declined between 2009 and 2016. The findings are concerning and represent a gap between treatment guidelines and clinical practice.Declaration of interestNone.


Indications for an expandable metallic stent for tracheobronchial stenosis.
An expandable metallic stent was used in 9 patients with tracheobronchial stenosis. Of the 8 patients with malignant stenosis, 6 had extrinsic compression and 2 had intraluminal tumor invasion. The 1 patient with nonmalignant stenosis had postreconstruction bronchomalacia and granulation tissue affecting different parts of the tracheobronchial tree. The expandable metallic stent successfully dilated tracheobronchial stenosis due to extrinsic tumor compression and malacia. However, it was not effective for stenosis due to intraluminal tumor invasion or granulation tissue because of the growth of tumor or granulation tissue between the wires of the stent. However, an expandable metallic stent covered with Dacron mesh was effective in 1 patient with intraluminal tumor invasion because tumor growth was blocked by the mesh. We conclude that an expandable metallic stent is effective for tracheobronchial stenosis due to extrinsic tumor compression, but not for stenosis due to intraluminal tumor invasion or granulation tissue.


Neuro-ophthalmic features of cerebral venous obstruction.
To characterize the neuro-ophthalmologic manifestations of impaired cranial venous outflow. A retrospective study of 20 patients who developed increased intracranial pressure as the result of impaired cerebral drainage. Three neuro-ophthalmologic referral centers. Ten patients had noncompressive thrombosis of sagittal or lateral sinuses (noncompressive group), four had compression of the sagittal sinus or jugular veins (compressive group), and six had transient sinus thrombosis following surgical procedures (iatrogenic group). Medical and/or surgical treatment of increased intracranial pressure and of underlying conditions predisposing to venous obstruction. Symptoms (headache and esodeviation) and signs (visual acuity, visual fields, and optic disc appearance) of increased intracranial pressure. Headache was more severe, esodeviation was more prevalent, and onset was more abrupt in the noncompressive group than in the compressive and iatrogenic groups. Among the 10 patients with noncompressive thrombosis, factors predisposing to thrombosis were present in seven patients but sometimes overlooked. Following treatment, visual outcome was generally favorable in all three groups. All patients had 20/30 or better visual acuity in at least one eye, and substantial visual field loss was present in both eyes of only two patients. The clinical manifestation of impaired cerebral venous outflow vary depending on the underlying mechanism. Although most patients maintain good vision, more aggressive treatment is sometimes indicated in patients with thrombosis or compression. The abrupt onset and marked severity of symptoms in patients with thrombosis should help to differentiate these patients from those with idiopathic intracranial hypertension.


Rebuilding for the future: a forward-looking master plan.
A 10-state redevlopment plan will provide a new structure with minimal interruption of service and loss of revenue.


The neuroprotective effect of hyperoxygenate hydrogen-rich saline on CO-induced brain injury in rats.
This study was designed to investigate the neuroprotective effect of hyperoxygenate hydrogen-rich saline (HOHS) against brain injury induced by carbon monoxide (CO) poisoning in rats. A rat model of CO poisoning was established by administering CO via intraperitoneal injection to male Sprague-Dawley rats. Forty-eight adult male rats were randomly divided into the following groups: normal control group (NG), CO poisoning group (CO), HOS treatment group (hyperoxygenated solution, HOS) and HOHS treatment group (HOHS). After CO poisoning, the carboxyhemoglobin (COHb) contents in the blood of rats in all the CO poisoning groups were increased significantly. However, HOS and HOHS significantly decreased COHb contents, furthermore, the HOHS group had lower COHb contents than the HOS group. Arterial oxygen partial pressure (PaO2) and arterial oxygen saturation (SaO2) results showed that HOS and HOHS could improve the oxygenation of the rats with CO poisoning. Compared with the CO group, the HOS group and the HOHS group had persistently neuroprotective effect on CO-induced brain injury, as assessed by modified neurological severity score (mNSS), furthermore, the HOHS group had better neurological functional recovery than the HOS group. The neuronal apoptosis induced by CO was also evaluated. Except the NG group, all the CO-poisoning groups had varying degrees of neuronal apoptosis. There was lesser degree of neuronal apoptosis in both the HOS group and the HOHS group than that in the CO group. Moreover, the HOHS group had more minor degree of neuronal apoptosis than the HOS group. Compared with the CO group, the free radicals production in the HOS group and the HOHS group were significantly inhibited. In addition, there were significantly difference in the free radicals production between the HOS group and the HOHS group. We could conclude that HOHS exerted a stronger neuroprotective effect against CO-induced brain injury than HOS, and the neuroprotective mechanism of HOHS may be related with inhibition of both neuronal apoptosis and free radicals.


Populus alba cationic cell-wall-bound peroxidase (CWPO-C) regulates the plant growth and affects auxin concentration in Arabidopsis thaliana.
The poplar cationic cell-wall-bound peroxidase (CWPO-C) mediates the oxidative polymerization of lignin precursors, especially sinapyl alcohols, and high molecular weight compounds that cannot be oxidized by other plant peroxidases, including horseradish peroxidase C. Therefore, CWPO-C is believed to be a lignification-specific peroxidase, but direct evidence of its function is lacking. Thus, the CWPO-C expression pattern in Arabidopsis thaliana (Arabidopsis) was determined using the β-glucuronidase gene as a reporter. Our data indicated that CWPO-C was expressed in young organs, including the meristem, leaf, root, flower, and young xylem in the upper part of the stem. Compared with the wild-type control, transgenic Arabidopsis plants overexpressing CWPO-C had shorter stems. Approximately 60% of the plants in the transgenic line with the highest CWPO-C content had curled stems. These results indicate that CWPO-C plays a role in cell elongation. When plants were placed horizontally, induced CWPO-C expression was detected in the curved part of the stem during the gravitropic response. The stem curvature associated with gravitropism is controlled by auxin localization. The time needed for Arabidopsis plants overexpressing CWPO-C placed horizontally to bend by 90° was almost double the time required for the similarly treated wild-type controls. Moreover, the auxin content was significantly lower in the CWPO-C-overexpressing plants than in the wild-type plants. These results strongly suggest that CWPO-C has pleiotropic effects on plant growth and indole-3-acetic acid (IAA) accumulation. These effects may be mediated by altered IAA concentration due to oxidation. The online version contains supplementary material available at 10.1007/s12298-022-01241-0.


Antigen delivery via hydrophilic PEG-b-PAGE-b-PLGA nanoparticles boosts vaccination induced T cell immunity.
Here, we evaluate the use of hydrophilic PEG-b-PAGE-b-PLGA (PPP) for the preparation of antigen loaded nanoparticles (NPs) as a platform for prophylactic vaccination. To investigate the suitability of PPP-NPs for antigen delivery, we used the double emulsion evaporation technique to prepare NPs of different sizes, antigen-loading efficiencies and -release kinetics for the model antigen Ovalbumin (OVA). Prior to applying the PPP-NPs in biological in vitro or in vivo models, all materials were tested for absence of cytotoxicity and endotoxins. While the uptake of NPs in antigen presenting cells was size but not polymer dependent, the efficiency of cross presentation of NP-associated antigen on MHC I molecules for CD8 T cell activation depended on the polymer type. T cell activation by antigen-presenting cells was significantly increased in vitro if antigen was delivered via PPP NPs compared to PLGA NPs or soluble OVA, although antigen content was the same in all tested formulations. Subcutaneous application of PPP-OVA-NPs even without adjuvants led to generation of potent CD8 T cell-mediated OVA-specific cytotoxicity in vivo that was more pronounced than after application of OVA alone or PLGA-OVA-NPs. Our data suggest that PPP-NPs can serve as platform for antigen-delivery in future vaccination formulations. Although PPP-NPs already bear intrinsic adjuvant-function, the complementation with TLR ligands loaded inside NPs may further strengthen the immune response to a point, where it might be possible to use it as a therapeutic vaccine to break immune tolerance in chronic disease states.


Blonanserin suppresses impulsive action in rats.
High impulsivity will increase the risk of criminal behavior, drug abuse, and suicide. We chose two drugs by following a strategy recently we proposed for identifying potential anti-impulsivity drugs, and examined the effects on impulsive action in rats by using a 3-choice serial reaction time task. We showed that the administration of blonanserin, an atypical antipsychotic, reduced impulsive actions in a U-shaped manner. 1-(2-Pyriidinyl)-piperazine, an active metabolite of buspirone or tandospirone, also slightly reduced impulsive actions, though it impaired motor functions. These results affirm the validity of our strategy, but require its refinement for developing anti-impulsivity drugs.


Sacred Practices and Family Processes in a Jewish Context: Shabbat as the Weekly Family Ritual Par Excellence.
The present article provides a deep and more focused look at the utility, meaning, processes, and power involved in a specific, family-level, sacred practice or ritual from Judaism: Shabbat (Sabbath). Content analysis of in-depth interviews with 30 diverse, marriage-based Jewish families living in the United States (N = 77 individuals) yielded three emergent themes: (a) "Shabbat brings us closer together"; (b) How Shabbat brings the family together; and (c) The Power of Blessing the Children. These themes will be discussed respectively, along with related verbatim data from participants' in-depth qualitative interviews.


Androgen Receptor Is Expressed in the Majority of Breast Cancer Brain Metastases and Is Subtype-Dependent.
We aimed to evaluate the expression of the "targetable" androgen receptor (AR) in breast cancer brain metastases (BrM). An established, retrospective 57-patient cohort with metastatic breast cancer who underwent surgery for BrM at the Sunnybrook Odette Cancer Centre between 1999-2013 was studied. AR expression in BrM samples was assessed in triplicate using immunohistochemistry (IHC). AR positive status was defined as nuclear AR expression ≥ 10% by IHC using the SP107 antibody. The median age of patients was 52 years (range 32-85 years). 28 (49%) of BrM were HER2+, 17 (30%) were hormone receptor positive (HR+)/HER2-, and 12 (21%) were triple negative breast cancers (TNBCs). 56% (n = 32/57) of BrM were AR positive, and median AR expression was 20% (CI 1.6-38.3%). AR expression was different across breast cancer subtypes; AR was most frequently expressed in HER2+ (n = 21/28), followed by HR+/HER2- (n = 9/17), and lowest in TNBC (n = 2/12) BrM (p = 0.003). Patients with AR positive versus AR negative BrM had similar overall survival (12.5 vs. 7.9 months, p = 0.6), brain-specific progression-free survival (8.0 vs. 5.1 months, p = 0.95), and time from breast cancer diagnosis to BrM diagnosis (51 vs. 29 months, p = 0.16). AR is expressed in the majority of breast cancer BrM and represents a potential therapeutic target.


Cortical brain activity in transfemoral or knee-disarticulation prosthesis users performing single- and dual-task walking activities.
Walking with a prosthesis while performing secondary tasks increases demand on cognitive resources, compromising balance and gait. This study investigated effects of a secondary task on patterns of brain activity and temporospatial gait parameters in individuals using a prosthesis with or without a microprocessor-controlled prosthetic knee(MPK) and controls. A cross-sectional study with repeated measures was performed. Twenty-nine individuals with amputations and 16 controls were recruited. Functional near-infrared spectroscopy was used to evaluate changes in oxygenated and de-oxygenated haemoglobin in the prefrontal cortex and temporospatial variables during single-and dual-task walking. Differences in brain activity were observed within the MPK-group and controls without changes in temporospatial parameters. The Trail-Walking test was associated with highest levels of brain activity in both groups. No differences were observed between single- and dual-task walking in the non-MPK-group (p > 0.05). The Non-MPK and the MPK-group recorded higher levels of brain activity than controls during single-task walking and poorer results on temporospatial variables compared to controls. For the MPK-group and controls, introduction of a secondary task led to an increase in brain activity. This was not seen in the Non-MPK-group. Significant differences in brain activity were observed in the absence of changes in temporospatial parameters.


Natural occurrence of ochratoxin A and citrinin in food stuffs in Egypt.
Natural occurrence of ochratoxin A (OA) and citrinin in cereals (274 samples) and animal tissues (250 samples) have been investigated during a period of more than 2 years. OA was found in cereals and animal tissues while citrinin was found in cereals only. The highest level of OA (up to 80.0 μg/kg) was found in yellow corn, 52.8% of contaminated samples while respectively 55.9% and 39.4% of barley and rice samples were contaminated with citrinin, with the highest level up to 100.0 and 27.92 μg/kg for barley and rice respectively. The frequent contamination of animal kidney with OA (28% positive out of 150 tested) average concentration 12.33 μg/kg. 2% of liver and 4% of muscles tissue were observed.


Applying a model of saccadic prediction to patients' saccadic eye movements.
A mathematical model of eye tracking of periodic square wave target motion has previously been proposed. In this work the model was further studied with regard to saccadic response time to symmetrical square wave target motion in 6 patients with unilateral parietal lesions following cerebrovascular accidents. Three types of saccadic response time were noted. In patients with type I, saccades contralateral to the damaged hemisphere had similar characteristics to those found in ipsilateral direction, but with longer response time. In those with type II, the saccadic response time was about the same on movement in either direction for repetitive target cycle durations of 1.2 or 2.0 s. In patients with type III, the target motion could not be predicted, in either direction.


Quantification of yessotoxin using the fluorescence polarization technique and study of the adequate extraction procedure.
Yessotoxin (YTX) is a polycyclic ether toxin produced by phytoplanktonic microalgae from the group of dinoflagellates. It has been shown that YTX increases the 3',5'-cyclic nucleotide phosphodiesterases (PDEs) activity and that there is a binding between these proteins and the toxin. Fluorescence polarization (FP) is a spectroscopic technique that can be used to study the interactions between molecules. It is based on exciting a fluorescent molecule with plane-polarized light and measuring the polarization degree of the emitted light. In this study, the FP is applied to the study of the interaction between YTX and phosphodiesterases I and II (PDE I and II). The phosphodiesterases are labeled with a reactive succinimidyl esther of carboxyfluorescein, and the FP of the protein-dye conjugate is measured when the YTX concentration in the medium increases. The results show that in both cases the fluorescence polarization of the conjugates decreases when they bind to YTX. For the PDE I, it is possible to draw a Gaussian curve or a straight line that relates the two variables (FP and YTX concentration). The concentration of this toxin in a spiked mussel extract (which contains the conjugate) can be quantified measuring its FP and using the equations of those lines. Different extraction methods are tried in this study, and those that can be used to obtain an appropriate mussel extract to be quantified with this technique are determined.


Noncontiguous finished genome sequence and description of Planococcus massiliensis sp. nov., a moderately halophilic bacterium isolated from the human gut.
We propose the main phenotypic characteristics and the complete genome sequence and annotation of Planococcus massiliensis strain ES2(T) (= CSUR P1103 = DSM 28915), the type strain of P. massiliensis sp. nov., isolated from a faeces sample collected from a healthy Senegalese man. It is an aerobic, Gram-positive, moderately halophilic, motile and rod-shaped bacterium. The 3 357 017 bp long genome exhibits a G+C content of 46.0% and contains 3357 protein-coding genes and 48 RNA genes.


Using 'draw, write and tell' to understand children's health-related experiences.
In recognising the capability and rights of children to express their experiences, 'draw, write and tell' (DWT) has emerged as a participatory qualitative research method. DWT enables children to communicate their experiences by drawing, writing words and telling the story of their pictures in response to interview questions. To discuss the challenges and benefits of using DWT to explore children's experiences of pain. Aspects that affect the quality of data in DWT include the materials used and the influences of the primary caregiver. Experience suggests that if trust between the child and researcher has been established, the duration of the interview is unimportant. While many methods of analysis can be used with data gathered using DWT, it is important to ensure children's perspectives are represented accurately. Furthermore, children's capacity as active participants in research should be reflected. Future studies could examine the potential of using drawings to share information in adult and paediatric clinical settings.


Annexin A protein family in atherosclerosis.
Atherosclerosis, a silent chronic vascular pathology, is the cause of the majority of cardiovascular ischaemic events. Atherosclerosis is characterized by a series of deleterious changes in cellularity, including endothelial dysfunction, transmigration of circulating inflammatory cells into the arterial wall, pro-inflammatory cytokines production, lipid accumulation in the intima, vascular local inflammatory response, atherosclerosis-related cells apoptosis and autophagy. Proteins of Annexin A (AnxA) family, the well-known Ca2+ phospholipid-binding protein, have many functions in regulating inflammation-related enzymes and cell signaling transduction, thus influencing cell adhesion, migration, differentiation, proliferation and apoptosis. There is now accumulating evidence that some members of the AnxA family, such as AnxA1, AnxA2, AnxA5 and AnxA7, play major roles in the development of atherosclerosis. This article discusses the major roles of AnxA1, AnxA2, AnxA5 and AnxA7, and the multifaceted mechanisms of the main biological process in which they are involved in atherosclerosis. Considering these evidences, it has been proposed that AnxA are drivers- and not merely participator- on the road to atherosclerosis, thus the progression of atherosclerosis may be prevented by targeting the expression or function of the AnxA family proteins.


Detection of Alzheimer's disease using features of brain region-of-interest-based individual network constructed with the sMRI image.
Brain networks constructed with regions of interest (ROIs) from the structural magnetic resonance imaging (sMRI) image are widely investigated for detecting Alzheimer's disease (AD). However, the ROI is generally represented by spatial domain-based features, so attentions are hardly paid to constructing a brain network with the frequency domain-based feature. In order to accurately characterize the ROI in the frequency domain and then construct an individual network, in this study, a novel method, which can describe the ROI properly by directional subbands and capture correlations between those ROIs, is proposed to construct a shearlet subband energy feature-based individual network (SSBIN) for AD detection. Specifically, the SSBIN is constructed with 90 ROIs which are segmented from the pre-processed sMRI image based on the automated anatomical labeling atlas, the 90 ROIs are represented by directional subband-based energy feature vectors (SVs) formed by jointing energy features extracted from their directional subbands, and the weight values of the SSBIN are computed by Pearson's correlation coefficient (PCC). Subsequently, two network features are extracted from the SSBIN: the node feature vector (NV) is computed by averaging the 90 SVs; the low dimensional edge feature vector (LV) is obtained by kernel principal component analysis (KPCA). Following that the concatenation of NV and LV is used as a SSBIN-based feature for the sMRI image. Finally, we use support vector machine (SVM) with the radial basis function kernel as classifier to categorize 680 subjects selected from the AD Neuroimaging Initiative (ADNI) database. Experimental results validate that the ROI can be properly characterized by the NV, and correlations between ROIs captured by the LV play an important role in AD detection. Besides, a series of comparisons with four current state-of-the-art approaches demonstrate the higher AD detecting performance of the SSBIN method.


A new Rothamsted long-term field experiment for the twenty-first century: principles and practice.
Agriculture faces potentially competing societal demands to produce food, fiber and fuel while reducing negative environmental impacts and delivering regulating, supporting and cultural ecosystem services. This necessitates a new generation of long-term agricultural field experiments designed to study the behavior of contrasting cropping systems in terms of multiple outcomes. We document the principles and practices of a new long-term experiment of this type at Rothamsted, established at two contrasting sites in 2017 and 2018, and report initial yield data at the crop and system level. The objective of the Large-Scale Rotation Experiment was to establish gradients of system properties and outcomes to improve our fundamental understanding of UK cropping systems. It is composed of four management factors-phased rotations, cultivation (conventional vs reduced tillage), nutrition (additional organic amendment vs standard mineral fertilization) and crop protection (conventional vs smart crop protection). These factors were combined in a balanced design resulting in 24 emergent cropping systems at each site and can be analyzed at the level of the system or component management factors. We observed interactions between management factors and with the environment on crop yields, justifying the systems level, multi-site approach. Reduced tillage resulted in lower wheat yields but the effect varied with rotation, previous-crop and site. Organic amendments significantly increased spring barley yield by 8% on average though the effect again varied with site. The plowed cropping systems tended to produce higher caloric yield overall than systems under reduced tillage. Additional response variables are being monitored to study synergies and trade-offs with outcomes other than yield at the cropping system level. The experiment has been established as a long-term resource for inter-disciplinary research. By documenting the design process, we aim to facilitate the adoption of similar approaches to system-scale agricultural experimentation to inform the transition to more sustainable cropping systems. The online version contains supplementary material available at 10.1007/s13593-023-00914-8.


Microstructural retinal findings by spectral-domain optical coherence tomography after vitrectomy repair of rhegmatogenous retinal detachments.
To evaluate macular microstructural changes after macula-involving rhegmatogenous retinal detachment repair and correlate with visual recovery. A prospective, observational study was performed of select patients presenting with macula-involving rhegmatogenous retinal detachments (RRD). Spectral-domain optical coherence tomography (SD-OCT) imaging was performed at postoperative visits over the course of the first year after operative repair of the RRD. The maculas of seven patients were evaluated. Postoperative best corrected visual acuity (BCVA) at the final follow-up ranged from 20/20 to 20/70 for all patients. Eyes with final BCVA greater than 20/40 had intact external limiting membrane and outer photoreceptor structures, while those with BCVA of less than 20/70 had poorer definition to those structures. Serial images showed resolution of irregularities in both the external limiting membrane and outer photoreceptor structures. Macular structure and function can progressively improve in the first year after successful RRD repair, including in the external limiting membrane, damage to which has previously been thought to be irreversible. SD-OCT serves as a useful tool to monitor postoperative retinal recovery.


Do community health workers affect non-urgent, ambulatory healthcare utilization among low-income, minority patients with diabetes?
Community health workers (CHWs) can reduce emergent care among low-income, ethnic minority patients with type 2 diabetes. A secondary analysis of a randomized controlled trial evaluated the effect of CHWs on non-urgent, ambulatory healthcare utilization. Within this trial, no effect on ambulatory care was found.


Further analysis on the structural proteins of infectious pancreatic necrosis virus.
The structural proteins of infectious pancreatic necrosis virus (IPNV) have been analyzed. Two-dimensional gel electrophoresis showed that IPNV proteins are slightly acidic with apparent pIs ranging from 5.8 to 6.6. To identify the IPNV surface-located proteins, purified virus was labelled either with fluorescein isothiocyanate (FITC) or with Na 125I. After analysis by SDS-PAGE, only the major viral protein, VP2, was labelled by either procedure. The accessibility of VP2 to these reagents suggests that this protein is externally located. In addition, using Concanavalin A conjugated with FITC and IPNV labelling with 3H-mannose, evidence is present that VP2 contains carbohydrate residues.


Pharmacological evidence for the presence of cholecystokinin-containing neurones in the mesenteric nerves supplying the guinea-pig ileum.
In guanethidine-treated, capsaicin-desensitized segments of guinea-pig isolated ileum electrical stimulation of mesenteric nerves resulted in contractions reaching 10 to 30% of the maximal longitudinal spasm. These responses were abolished by tetrodotoxin or atropine, and were reduced by half by the selective cholecystokinin antagonist lorglumide. It is concluded that neuronal processes whose endings release cholecystokinin-like material within the gut wall run in the mesenteric nerves and contribute to the capsaicin-resistant contractile response by releasing endogenous acetylcholine.


Erythropoietin to minimize perioperative blood transfusion: a systematic review of randomized trials. The International Study of Peri-operative Transfusion (ISPOT) Investigators.
Our aim was to perform a systematic review to determine the efficacy and side-effects of erythropoietin, given with or without autologous predonation, to patients undergoing orthopaedic or cardiac surgery. A number of studies have been done to determine whether erythropoietin minimizes exposure to perioperative allogeneic red cell transfusion. A systematic review of all randomized trials will provide the best estimate of the efficacy and side-effects of erythropoietin therapy. All randomized trials of erythropoietin in cardiac or orthopaedic surgery that reported the proportion of patients receiving perioperative allogeneic transfusion were included. The efficacy of erythropoietin was evaluated in subgroups of patients depending upon the route of administration, dose of erythropoietin, the type of control and the methodological quality of the study report. The odds ratio for the proportion of patients transfused with allogeneic blood in studies of erythropoietin to augment autologous donation was 0.42 (95% confidence limits 0.28-0.62; P < 0.0001) for orthopaedic surgery and 0.25 (95% CI 0.08-0.82; P = 0.02) for cardiac surgery. The odds ratio for erythropoietin alone was 0.36 (95% CI 0.24-0.56; P = 0.0001) in orthopaedic surgery and 0.25 (95% CI 0.06-1.04; P < 0.06) in cardiac surgery. The route of administration, dose of erythropoietin, type of control and methodological quality of the study report had no statistically significant effect upon the odds ratios. Although there was no convincing evidence that erythropoietin used alone increased the frequency of thrombotic complications, some studies found an excess of events in erythropoietin-treated patients, and the number of patients studied was relatively small. Erythropoietin, when given alone or to augment autologous donation, decreased exposure to perioperative allogeneic transfusion in orthopaedic and cardiac surgery. Further studies are required to definitively establish the safety of erythropoietin alone, to determine the optimal dose of perioperative erythropoietin, and to compare its efficacy and cost-effectiveness with other methods of minimizing perioperative transfusion.


An early outward conductance modulates the firing latency and frequency of neostriatal neurons of the rat brain.
An in vitro slice preparation was used to obtain intracellular recordings of neostriatal neurons. Indirect evidence for the presence of an early outward conductance in neostriatal neurons is presented. With near threshold stimulation neostriatal neurons fired very late during the pulse. The long firing latency was associated with a slow (ramp-like) depolarization. In the presence of TTX the slow depolarization was lost and outward-going rectification dominated the subthreshold response. This finding demonstrated that both, outward- and inward-going conductances play a role during the ramp-like depolarization. Outward-going rectification during depolarizing responses could be further augmented if the depolarizing stimulus was preceded by a conditioning hyperpolarization. A conditioning hyperpolarization prolonged the firing latency and slowed the firing frequency. A conditioning depolarization had opposite effects. After TTX treatment, the response showed a hyperpolarizing "sag" when depolarizing stimulation was preceded by conditioning hyperpolarization. 4-AP (0.5-2.5 mM) blocked the effects of the conditioning hyperpolarization on the firing latency and on the voltage trajectory. 4-AP also disclosed a slow depolarization which could produce neuronal firing very early during the pulse. This depolarization was TTX-sensitive and Co++-insensitive. In contrast to 4-AP, TEA (20 mM) did not produce a reduction in the firing latency but disclosed a membrane oscillatory behavior most probably produced by the interplay of these opposing conductances: the slow inward (probably Na+) and the transient outward (probably K+). Repetitive firing during 4-AP treatment was of the "phasic-tonic" type with an initial burst riding on the initial Co++-insensitive slow depolarization and a somehow irregular train of spikes during the remainder of the stimulation. Action potentials during 4-AP treatment were followed by an afterdepolarization which dominated the initial part of the interspike interval.


Homology-dependent DNA transfer from plants to a soil bacterium under laboratory conditions: implications in evolution and horizontal gene transfer.
DNA transfer was demonstrated from six species of donor plants to the soil bacterium, Acinetobacter spp. BD413, using neomycin phosphotransferase (nptII) as a marker for homologous recombination. These laboratory results are compatible with, but do not prove, DNA transfer in nature. In tobacco carrying a plastid insertion of nptII, transfer was detected with 0.1 g of disrupted leaves and in oilseed rape carrying a nuclear insertion with a similar quantity of roots. Transfer from disrupted leaves occurred in sterile soil and water, without the addition of nutrients. It was detected using intact tobacco leaves and intact tobacco and Arabidopsis plants in vitro. Transfer was dose-dependent and sensitive to DNase, and mutations in the plant nptII were recovered in receptor bacteria. DNA transfer using intact roots and plants in vitro was easily demonstrated, but with greater variability. Transfer varied with plant genome size and the number of repeats of the marker DNA in the donor plant. Transfer was not detected in the absence of a homologous nptII in the receptor bacteria. We discuss these results with reference to non-coding DNA in plant genomes (e.g., introns, transposons and junk DNA) and the possibility that DNA transfer could occur in nature.


Tree-Based Analysis of Dietary Diversity Captures Associations Between Fiber Intake and Gut Microbiota Composition in a Healthy US Adult Cohort.
Diet patterns are a significant and modifiable contributing factor to the composition of the human gut microbiota. We set out to identify reproducible relationships between diet and gut microbial community composition in a diverse, healthy US adult cohort. We collected 2 to 3 automated self-administered 24-hour dietary recalls over 10-14 days, together with a single stool sample, from 343 healthy adults in a cross-sectional phenotyping study. This study examined a multi-ethnic cohort balanced for age (18-65 years), sex, and BMI (18.5-45 kg/m2). Dietary data were edited to a tree format according to published methods. The tree structure was annotated with the average total grams of dry weight, fat, protein, carbohydrate, or fiber from each food item reported. The alpha and beta diversity measurements, calculated using the tree structure, were analyzed relative to the microbial community diversity, determined by a Quantitative Insights Into Microbial Ecology (QIIME) 2 analysis of the bacterial 16S ribosomal RNA V4 region, sequenced from stool samples. K-means clustering was used to form groups of individuals consuming similar diets, and gut microbial communities were compared among groups using differential expression analysis for sequence count data. The alpha diversity of diet dry weight was significantly correlated with the gut microbial community alpha diversity (r = 0.171). The correlation improved when diet was characterized using grams of carbohydrates (r = 0.186) or fiber (r = 0.213). Bifidobacterium was enriched with diets containing higher levels of total carbohydrate from cooked grains. Lachnospira, was enriched with diet patterns containing high consumption of fiber from fruits excluding berries. The tree structure, annotated with grams of carbohydrate, is a robust analysis method for comparing self-reported diet to the gut microbial community composition. This method identified consumption of fiber from fruit robustly associated with an abundance of pectinolytic bacterial genus, Lachnospira, in the guts of healthy adults. This trial was registered at clinicaltrials.gov as NCT02367287.


[Ultrastructure of reparation collagen and its relations to residual hyaline joint cartilage in inflammatory cartilage destruction].
The development of collagenous fibrils in reparative tissues replacing hyaline cartilage in arthritic joints was investigated by electron microscopy: granulation tissue consists of a loose network of collagenous fibrils, scar tissue is characterized by dense bundles of parallel arranged collagenous fibrils, chondroid tissue that may develop under certain instances is characterized by a network of random orientated fibrils that differ from those of the normal hyaline cartilage by their uniformity. The fibrils of the reparative tissues often exhibited an intense contact with those of the original hyaline cartilage.


Methodological comparison of FACS and MACS isolation of enriched microglia and astrocytes from mouse brain.
Microglia and astrocytes, the two innate cells in CNS, are thought to protect and remodel of synapses for proper maintenance and plasticity of neuronal circuits. The two types of cells are the major responders by producing and releasing inflammatory mediators. Isolation of microglia and astrocytes from CNS tissue provides a powerful tool to study basic cell biology and examine the effects of in vivo treatments on microglia and astrocytes immunophenotype and function. The widely used approach of enrichment microglia and astrocytes from CNS was MACS (Magnetic activated cell sorting) and FACS (Fluorescence activated cell sorting). Here we described an optimized protocol of enzymatic dissociation generating single cell suspensions from brain tissue. Then the ability of the two methods to isolate microglia and astrocytes from brain dissociated cells was compared. Both MACS and FACS processing could obtain microglia and astrocytes with high viability (>85%). Microglia sorted by MACS comprises a slight myeloid cells contamination but with a little bit higher efficiency than that sorted by FACS. MACS processing was faster than FACS for either single or multiple samples. ACSA2 can be used to isolate astrocytes from both postnatal and adult brain, and is more suitable for purify astrocytes from newborn. FACS could get purer microglia which is helpful for deep sequencing and other related research. ACSA2 is a good marker of astrocytes.


Dystonic reaction associated with ondansetron administration in a patient with normal pressure hydrocephalus.
A 77-year-old female with normal pressure hydrocephalus underwent urgent revision of the abdominal component of a ventriculoperitoneal shunt. Upon emergence from an uneventful general anesthetic, the patient exhibited cogwheel rigidity with decerebrate posturing and a markedly irregular respiratory rate. This prohibited extubation and prompted an unplanned intensive care unit admission. Ondansetron was the only medication administered that had previously been associated with dystonic reactions. In a previous procedure, the patient had been given ondansetron without a similar reaction. This presentation may have been provoked by existing pathology such as her underlying normal pressure hydrocephalus and small increases in intracranial pressure.


Identification of CD25 as STAT5-Dependent Growth Regulator of Leukemic Stem Cells in Ph+ CML.
In chronic myelogenous leukemia (CML), leukemic stem cells (LSC) represent a critical target of therapy. However, little is known about markers and targets expressed by LSCs. The aim of this project was to identify novel relevant markers of CML LSCs. CML LSCs were examined by flow cytometry, qPCR, and various bioassays. In addition, we examined the multipotent CD25(+)CML cell line KU812. In contrast to normal hematopoietic stem cells, CD34(+)/CD38(-)CML LSCs expressed the IL-2 receptor alpha chain, IL-2RA (CD25). STAT5 was found to induce expression of CD25 in Lin(-)/Sca-1(+)/Kit(+)stem cells in C57Bl/6 mice. Correspondingly, shRNA-induced STAT5 depletion resulted in decreased CD25 expression in KU812 cells. Moreover, the BCR/ABL1 inhibitors nilotinib and ponatinib were found to decrease STAT5 activity and CD25 expression in KU812 cells and primary CML LSCs. A CD25-targeting shRNA was found to augment proliferation of KU812 cellsin vitroand their engraftmentin vivoin NOD/SCID-IL-2Rγ(-/-)mice. In drug-screening experiments, the PI3K/mTOR blocker BEZ235 promoted the expression of STAT5 and CD25 in CML cells. Finally, we found that BEZ235 produces synergistic antineoplastic effects on CML cells when applied in combination with nilotinib or ponatinib. CD25 is a novel STAT5-dependent marker of CML LSCs and may be useful for LSC detection and LSC isolation in clinical practice and basic science. Moreover, CD25 serves as a growth regulator of CML LSCs, which may have biologic and clinical implications and may pave the way for the development of new more effective LSC-eradicating treatment strategies in CML.


Preparation and purification of estriol, estradiol, and progesterone labeled with radioactive iodine.
Steroids labeled with radioactive isotopes are useful tracers for radio-immunoassay and for in vivo nuclear medecine purposes. We here report and discuss methods for direct labeling of the steroids estriol, estradiol, and progesterone with radioiodine-125 or -131. The purification procedure and some chemical and physical properties of the purified labeled materials are referred to and discussed.


Measurement of 5-hydroxytryptophol and 5-hydroxyindoleacetic acid in human and rat brain and plasma.
The levels of 5-hydroxyindoleacetic acid (5-HIAA) and free and total 5-hydroxytryptophol (5-HTOL) in human and rat brain regions and plasma were determined by a specific capillary column gas chromatographic--mass spectrometric method. The human brains were obtained 2-3 hours post mortem, and the levels of 5-HIAA were in the range of 0.48-31.3 nmoles/g in the regions investigated. The levels of free and total 5-HTOL were 10.9-387 pmoles/g and 14.5-821 pmoles/g, respectively. The ratio of total 5-HTOL to 5-HIAA was in the range of 0.6-5.5%. In human plasma the levels of free and total 5-HTOL were 0.9 +/- 0.3 and 2.9 +/- 0.8 pmoles/ml +/- S.E.M., respectively. In regions of rat brain, the 5-HIAA levels ranged from 0.37-2.84 nmoles/g. Free and total 5-HTOL were in the range of 11.4-56.1 and 16.2-77.1 pmoles/g, respectively. The ratio of total 5-HTOL and 5-HIAA ranged from 2.3-5.1%. Higher levels of 5-HIAA and 5-HTOL occurred in the rat pineal gland. In rat plasma the levels of free and total 5-HTOL were 1.34 +/- 0.06 and 21.6 +/- 1.6 pmoles/ml +/- S.E.M., respectively.


His electrogram alternans (Zhang's phenomenon) and a new model of dual pathway atrioventricular node conduction.
In contrast to the current textbook model and the current clinical index of dual pathway atrioventricular (AV) nodal conduction, here we summarize the discovery and validation of Zhang's phenomenon (originally His electrogram alternans) as a new index of dual pathway conduction. We also describe the new findings of transverse-versus-longitudinal electrical propagation within the AV node as the electrophysiological basis underlining this new index. Thus, a new index and a new model of dual pathway AV conduction are being developed. We have reviewed current literature and provided evidence supporting a new index and a new model of dual pathway AV conduction. Recent data revealed that during fast pathway conduction, electrical excitation in the AV node propagates in a superior to inferior direction across AV conduction axis and fiber orientation to reach first the superior His bundle fibers. However, this transverse conduction can fail easily within the superior nodal domain at fast rates. The failing of transverse propagation permits electrical excitation formed at the posterior/inferior nodal region to propagate longitudinally along fiber orientation in a posterior to anterior direction through the inferior nodal domain to reach the inferior His bundle (slow pathway conduction). This transverse-versus-longitudinal electrical propagation within the AV node results in a functional dissociation in the distal node and formation of dual inputs into the His bundle, providing the electrophysiological basis for the formation of Zhang's phenomenon (His electrogram alternans). Based on strong experimental data, a new index and a new model of dual pathway AV nodal conduction are emerging, although they are still awaiting clinical validation.


Contaminations of metal in tissues of Siberian gull Larus heuglini: gender, age, and tissue differences.
The objective of the present study was to investigate the levels of metals, namely cadmium, lead, and zinc, in Siberian gull (Larus heuglini) (n = 15), in order to: (1) examine the sex and gender related variation in trace metal accumulation, and (2) to determine the significant between metal concentrations in the kidney, liver, and pectoral muscle. The concentrations were different between the tissues of bird as well as among the interaction (sex × age), but this difference (except cadmium in liver and zinc in kidney) between the gender (male and female) and age (adult and juvenile) didn't exist. Results showed that the metal concentrations in the Siberian gull were decreased in sequence of kidney > liver > muscle. The cadmium, lead, zinc concentration overall means they were measured as 2.2 ± 0.7, 8.8 ± 2.5, and 91.1 ± 37. 1 μg/g for kidney, 1.1 ± 0.2, 5.1 ± 0.8, and 68.3 ± 27.8 μg/g for liver, and 0.8 ± 0.1, 3.4 ± 0.6, and 34.4 ± 23.2 μg/g for pectoral muscle, respectively.