Air pollution and hospital outpatient visits for conjunctivitis: a time-series analysis in Tai'an, China.
Conjunctivitis is one of the most common eye-related health problems and significantly influences patients' quality of life. Whether air pollution increased the risks of conjunctivitis is still unclear. Daily counts of outpatient visits for conjunctivitis, air pollution, and meteorological data during January 1, 2015-December 31, 2019 were collected from Tai'an, China. Generalized additive model with Poisson distribution was used to estimate the relationship between air pollution and visits for conjunctivitis, after controlling for the long-term and seasonal trends, weather variables, and day of the week. The effect of air pollution on visits for conjunctivitis was generally acute and significant at the current day and disappeared after 2 days. The relative risk of conjunctivitis visits associated with per 10 μg/m3 increases in PM2.5, PM10, SO2, and NO2 at lag 0-2 days was 1.006 (95% CI: 1.001-1.011), 1.003 (95% CI: 1.000-1.0107), 1.023 (95% CI: 1.009-1.037), and 1.025 (95% CI: 1.010-1.040), respectively. The impact of air pollution on visits for conjunctivitis varied greatly by individual characteristics. The impact of NO2 was higher in males than in females, with the opposite trend for SO2 and PM2.5. Effect estimates of air pollutants were higher among return visits for conjunctivitis, the elderly, and white-collar workers. Our study highlights that the vulnerable subpopulations should pay more attention to protect themselves from air pollution.


Rapid Makerspace Microfabrication and Characterization of 3D Microelectrode Arrays (3D MEAs) for Organ-on-a-Chip Models.
Integrated sensors in "on-a-chip" in vitro cellular models are a necessity for granularity in data collection required for advanced biosensors. As these models become more complex, the requirement for the integration of electrogenic cells is apparent. Interrogation of such cells, whether alone or within a connected cellular framework, are best achieved with microelectrodes, in the form of a microelectrode array (MEA). Makerspace microfabrication has thus far enabled novel and accessible approaches to meet these demands. Here, resin-based 3D printing, selective multimodal laser micromachining, and simple insulation strategies, define an approach to highly customizable and "on-demand" in vitro 3D MEA-based biosensor platforms. The scalability of this approach is aided by a novel makerspace microfabrication assisted technique denoted using the term Hypo-Rig. The MEA utilizes custom-defined metal microfabricated microelectrodes transitioned from planar (2D) to 3D using the Hypo-Rig. To simulate this transition process, COMSOL modeling is utilized to estimate transitionary forces and angles (with respect to normal). Practically, the Hypo-Rig demonstrated a force of ~40N, as well as a consistent 70° average angular transitionary performance which matched well with the COMSOL model. To illustrate the scalability potential, 3 × 3, 6 × 6, and 8 × 8 versions of the device were fabricated and characterized. The 3D MEAs, demonstrated impedance and phase measurements in the biologically relevant 1 kHz range of 45.4 kΩ, and -34.6° respectively, for polystyrene insulated, ~70μm sized microelectrodes.


Transition state isosteres of the gamma-glutamyl peptide bond hydrolysis: synthesis and characterization of the psi (CH2NH) pseudopeptide analogue of glutathione.
The fully deprotected glutathione analogue containing the aminomethylene unit as transition state isostere of the gamma-Glu-Cys peptide bond was synthesized for the first time and characterized in both the reduced and oxidized forms.


Observation of the Kondo effect in a spin-3/2 hole quantum dot.
We report the observation of Kondo physics in a spin-3/2 hole quantum dot. The dot is formed close to pinch-off in a hole quantum wire defined in an undoped AlGaAs/GaAs heterostructure. We clearly observe two distinctive hallmarks of quantum dot Kondo physics. First, the Zeeman spin splitting of the zero-bias peak in the differential conductance is independent of the gate voltage. Second, this splitting is twice as large as the splitting for the lowest one-dimensional subband. We show that the Zeeman splitting of the zero-bias peak is highly anisotropic and attribute this to the strong spin-orbit interaction for holes in GaAs.


Profiling of human stratum corneum ceramides by means of normal phase LC/APCI-MS.
The ceramides of the stratum corneum are critical to maintaining the epidermal barrier function of the skin. A number of skin diseases and disorders are known to be related to impairments of the ceramide pattern. Therefore, obtaining mass spectrometric profiles of the nine ceramide classes known to exist aids our understanding of the underlying molecular mechanisms, which should eventually lead to new diagnostic opportunities: for example, the mass spectrometric profiles of patients suffering from serious skin diseases such as atopic dermatitis and psoriasis can be compared to those of healthy controls. Previous work on mass spectrometric analysis of ceramides relied mostly on GC/MS after hydrolysis and derivatization. The introduction of ESI-MS and LC/ESI-MS has provided new options for directly analyzing intact ceramides. However, some of the ceramide classes are not accessible to ESI-MS. However, as shown in this work, these limitations of GC/MS and ESI-MS can be overcome using a new approach based on normal phase LC interfaced with APCI-MS. Separation and online detection of the stratum corneum ceramide classes became possible in one run. Ceramide species with C26 and/or C28 fatty acid chains were the most abundant ones in Cer [NP], Cer [NH], Cer [AP], and Cer [AH]. The main component of Cer [AS] was C16. The omega-esterified ceramide classes Cer [EOS], Cer [EOP] and Cer [EOH] contained mostly species with fatty acids >C30. This was also the case for Cer [NS], suggesting an analogy to the omega-esterified ceramides. In addition, evidence for a new ceramide class Cer [NdS] was found.


Molecular insights into formation of nitrogenous disinfection byproducts from algal organic matter in UV-LEDs/chlorine process based on FT-ICR analysis.
Eutrophication is a globally concerned issue, which brings algal cells and algal organic matter (AOM) into drinking water treatment plants. AOM is an important branch of nitrogenous disinfection byproduct (N-DBP) precursors. The variation of AOM composition in UV-LEDs/chlorine process, and its relationship with N-DBP formation still remain much uncertainty. Herein, we used fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) to investigate AOM transformation in UV-LEDs/chlorine process, with UV285 and UV365 as light source, and screen for typical precursors of N-DBPs. We found that more nitrogen-containing compounds were generated after UV-LEDs/chlorine process, leading to the larger formation of N-DBPs in postchlorination. Compounds such as lignin, proteins, and amino sugars tends to be oxidized by reactive species in UV-LEDs/chlorine process. Further, compounds with higher O/C and higher weighted average double bond equivalence (DBEw) are easier to form N-DBPs, including dichloroacetonitrile and trichloronitromethane. Also, influence factors including pH, UV fluence, post-chlorination time and bromide concentration on N-DBP formation were evaluated. The results show that N-DBP formation generally followed the order of UV285/chlorine-postchlorination, UV365/chlorine-postchlorination, and direct chlorination. Our study provides comprehensive information on N-DBP formation from AOM in UV-LEDs/chlorine-postchlorination from molecular levels.


Supporting tuberculosis program in active contact tracing: a case study from Pakistan.
Tuberculosis (TB) is on the rise in Pakistan and there could be multiple reasons including poverty, difficulty in access to TB treatment services, non-compliance with treatment, social stigma etc. According to the TB program managers, limited treatment and testing sites for tuberculosis and lack of trained human resources play a major role in compromising TB management. A major lacuna in the TB control program is the absence of active contact tracing strategy. This is essential for a disease where positive cases are known to be able to infect a further 10‒15 individuals in a year. Tackling tuberculosis in Pakistan has been beleaguered by funding challenges and other systems' bottlenecks such as lack of skilled human resources and insufficient supply of medicines, despite the fact that disease burden is one of the highest in the world. Although it is a notifiable disease, active case finding, contact tracing and reporting is notoriously low throughout the country. Access to diagnostics and treatment facilities has been limited and stigma attached to the disease remains deeply entrenched among the communities. Researchers have shown that enhanced and active approaches to contact investigation effectively identifies additional patients with TB among household contacts at a relatively modest cost. USAID's Integrated Health Systems Strengthening and Service Delivery Activity extended support to the Health Departments of Sindh and Khyber Pakhtunkhwa provinces. In collaboration with the two provincial TB programs, community based active contact tracing was conducted on 17,696 individuals, based on the index cases. Among the contacts traced, 243 cases were diagnosed as drug sensitive or drug resistant TB. Awareness sessions were conducted to sensitize people on the various aspects of disease and importance of getting tested. The project also supported establishing three satellite Programmatic Management of Drug Resistant Tuberculosis (PMDT) sites for drug resistant TB treatment, enhancing the programs' diagnostic and testing capacity.


Vocabulary Diversity in Personal Narratives Produced in Response to the Global TALES Protocol in Dutch-Speaking Students with and without Dyslexia.
This study examines whether there are differences in expressive vocabulary between participants with and without dyslexia in personal narratives in response to the Global TALES protocol. 22 monolingual Dutch-speaking participants aged 11-16 with dyslexia and 22 age and gender-matched peers without dyslexia were assessed on measures of decoding, reading comprehension, and spelling of words, pseudowords, verbs, and sentences. The participants also produced personal narratives in response to the six prompts contained in the Global TALES protocol. We analyzed the personal narratives for expressive vocabulary and counted the total number of different words (TNDW). The study revealed a significant relationship between TNDW and reading comprehension (r = 0.45, p = 0.002, BF10 = 17.70), spelling words (r = 0.42, p = 0.005, BF10 = 8.93), and spelling and writing conventions in sentences (r = 0.37, p = 0.016, BF10 = 3.11). The Global TALES protocol was successful in eliciting personal narratives in the Dutch-speaking participants with and without dyslexia. Participants with dyslexia used fewer different words (M = 192.27, SD = 64.37; 95% CI: [151.84-232.71]) compared to peers without dyslexia (M = 265.50, SD = 116.28; 95% CI: [225.06-305.93]; F(1, 42) = 6.68; p = 0.013; η2 = 0.14). When we compared the probability of models, Bayesian factors revealed moderate evidence for group differences in TNDW (BF = 3.94). Our findings indicate that older school-age participants with dyslexia may lag behind their peers in expressive vocabulary in a personal narrative discourse task that is relevant to everyday functioning. The results of this study highlight the relationship between expressive vocabulary and reading comprehension and the importance of the assessment of spoken language skills in children with dyslexia. Reading problems might lead to less advanced spoken language, which in turn may negatively affect the expressive vocabulary growth in individuals with dyslexia.


Knee biomechanics during popular recreational and daily activities in older men.
Physical activity is recommended for older adults, including those with knee pathology. However, demands on the knee during popular recreational activities are unclear. The study purpose was to determine knee biomechanics in healthy older men during golf and bowling and compare them to activities of daily living. Three-dimensional motion analysis was used to determine knee biomechanics in 19 healthy males (45-73 years): 11 golfers and eight bowlers. Subjects performed walking, stair ascent, stair descent, and either golf or bowling. Comparisons were made between the recreational activity and activities of daily living. During bowling, flexion angle at peak extensor moment was as high as during stair descent, and peak extensor moment was as high as during stair ascent. For the golf lead knee, flexion angle at peak extensor moment and peak extensor moment were as high as during stair ascent, and peak abduction moment, internal and external rotation angles were larger than during all activities of daily living. Peak external rotation angle for the golf trail knee was larger than all activities of daily living. The greatest challenge for the knee of healthy older males during bowling is eccentric control of knee flexion. Golf poses challenges in all three planes of motion for the lead knee and in the transverse plane for the trail knee. Comparing mechanical demands on the knee during bowling and golf to those of stair negotiation provides a reference for clinicians when recommending recreational activities for older adults with knee pathology.


The prognostic value of cell surface antigens in low grade, non-invasive, transitional cell carcinoma of the bladder.
Tumors in 23 patients who presented with a low grade, non-invasive transitional cell carcinoma of the bladder were studied for blood group antigens A, B or O on the cell surface. Of 14 patients without cell surface antigens initially 13 suffered an invasive tumor subsequently and 1 had diffuse carcinoma in situ. Of 9 patients with cell surface antigens initially 8 did not have an invasive recurrence during a 5 or more-year followup and 1 did. The presence or absence of blood group cell surface antigen on a low grade, non-invasive transitional cell carcinoma of the bladder would seem to have value in predicting future recurrence with muscle invasion.


Characterization of the C-terminal region of molecular forms of human milk bile salt-stimulated lipase.
Bile salt-stimulated lipase (BSSL) in human milk exists in multiple molecular forms and it has been shown that approximately one-third of lactating mothers secrete two forms. to determine the structural features of BSSL that may give rise to this heterogeneity. Oligosaccharides present in the proline-rich region in the C-terminus of BSSL were investigated using deglycosylating enzymes and lectin affinity probing to determine the origin of the multiple molecular forms. It was found that the variability in the molecular mass of BSSL is due predominantly to glycosylation. The molecular forms contain similar sugar chains; all forms possess the core disaccharide Galbeta1-3GalNAc and beta-D-galactose, fucose linked at alpha1-6 and sialic acid linkage alpha2-3 to galactose. The molecular mass difference in the BSSL molecular forms cannot be attributed to the type of carbohydrate moiety in the sugar chains of the N- and O-linked sites suggesting that the differences arise from the extent or quantity of glycosylation. The oligosaccharides in the C-terminal region contain Lewis x and b and, less prominently, Lewis a antigenic structures. Owing to the presence of these blood-group-related antigenic determinants, the C-terminal region of BSSL may have an adhesive function in cell-cell interactions.


Method for the assessment of neuromuscular integrity and burrowing choice in vermiform animals.
The study of locomotion in vermiform animals has largely been restricted to animals crawling on agar surfaces. While this has been fruitful in the study of neuronal basis of disease and behavior, the reduced physical challenge posed by these environments has prevented these organisms from being equally successful in the study of neuromuscular diseases. Our burrowing assay allowed us to study the effects of muscular exertion on locomotion and muscle degeneration during disease (Beron et al., 2015), as well as the natural burrowing preference of diverse Caenorhabditis elegans strains (Vidal-Gadea et al., 2015). We describe a simple, rapid, and affordable set of assays to study the burrowing behavior of nematodes and other vermiform organisms which permits the titration of muscular exertion in test animals. We show that our burrowing assay design is versatile and can be adapted for use in widely different experimental paradigms. Previous assays for the study of neuromuscular integrity in nematodes relied on movement through facile and homogeneous environments. The ability of modulating substrate density allows our burrowing assay to be used to separate animal populations where muscular fitness or health are not visible differentiable by standard techniques. The simplicity, versatility, and potential for greatly facilitating the study of previously challenging neuromuscular disorders makes this assay a valuable addition that overcomes many of the limitations inherent to traditional behavioral tests of vermiform locomotion.


Identifying, rating, and remembering environmental sound events.
Sound events are sequences of closely grouped and temporally related environmental sounds that tell a story or establish a sense of place. The goal of our project was to create a set of sound events depicting various scenarios (such as a car accident, cooking breakfast, and walking outdoors) and to gather normative data about how people understand them. Samples of college students listened to 22 sound events over headphones in three self-paced, computer-based studies. In the Identification Task, 43 participants used text boxes to type descriptions of what was happening in the sound events. In the Rating Task, 39 participants used Likert scales to rate the sound events on the attributes of familiarity, complexity, and pleasantness. In the Memory Task, 42 participants answered two multiple-choice questions immediately after listening to each sound event. Detailed tables are provided for the following: (1) Description of the sound events and their components; (2) accuracy and response time measurements for each of the 22 sound events across the three studies; and (3) rank-orderings of the sound events by ease of identification, recognition of details, and rated familiarity, complexity, and pleasantness. Digital files of the stimuli, which may be of interest to auditory cognition researchers and clinical neuropsychologists, may be downloaded from either www.psychonomic.org/archive or www.cofc.edu/-marcellm/sound event studies/sndevent.htm.


MiR-199a-5p regulates rat liver regeneration and hepatocyte proliferation by targeting TNF-α TNFR1/TRADD/CASPASE8/CASPASE3 signalling pathway.
Abnormally expressed miR-199a-5p (miR-199a) has been frequently reported in multiple types of malignancies. Nevertheless, its effect in liver regeneration (LR) is largely still unclear. Herein, we investigated the function of miR-199a in hepatocyte proliferation during LR. As a result, miR-199a expression was significantly increased 12-30 h, in rat hepatic tissue, after partial hepatectomy (PH). The down-regulated expression of miR-199a inhibited proliferation as well as promoted cell apoptosis of BRL-3A. Additionally, TNF-α was found as a target of miR-199a. The administration of TNF-α siRNA regulated the effects of miR-199a on hepatocyte proliferation as well as miR-199a-modulated TNF-α/TNFR1/TRADD/CASPASE8/CASPASE3 signalling pathways. Taken together, these present findings suggested that miR-199a promoted hepatocyte proliferation as well as LR via targeting TNF-α/TNFR1/TRADD/CASPASE8/CASPASE3.


Calcium currents in acutely isolated stellate and pyramidal neurons of rat entorhinal cortex.
Calcium currents were studied in morphologically identified pyramidal and stellate neurons acutely isolated from layer II/III of rat entorhinal cortex, using the whole-cell patch-clamp configuration. The peak amplitude of high-voltage activated current (HVA) measured at +10 mV was not different in both neuron populations with 0.94+/-0.08 nA for pyramidal and 1.03+/-0.08 nA for stellate cells. Stellate neurons had a larger capacitance (14.4+/-1. 1 pF) than pyramidal neurons (9.6+/-0.8 pF), indicating a 50% larger cell surface. Most striking was the difference between the current density in stellate (79+/-8 pA/pF) versus pyramidal neurons (113+/-13 pA/pF). The potential of half maximal inactivation was not different: -37+/-2 mV (pyramidals) and -37+/-3 mV (stellates). Half of the cells contained a low-voltage activated calcium current (LVA) with a peak amplitude that was twice as large in stellate as in pyramidal neurons (0.21+/-0.04 nA resp. 0.11+/-0.03 nA; at -50 mV). In contrast to the HVA component, the current density of the LVA component was not different between cell types (13+/-3 pA/pF vs. 13+/-2 pA/pF). This implies that the relative abundance of LVA and HVA currents in stellate and pyramidal neurons is different which could result in different firing characteristics. The potential of half maximal LVA inactivation was -88+/-4 mV (pyramidals) and -85+/-3 mV (stellates). The slope of the voltage dependent steady state inactivation was steeper in stellate (7+/-1 mV) than in pyramidal cells (10+/-2 mV).


Somatic Arc protein expression in hippocampal granule cells is increased in response to environmental change but independent of task-specific learning.
Activated neurons express immediate-early genes, such as Arc. Expression of Arc in the hippocampal granule cell layer, an area crucial for spatial learning and memory, is increased during acquisition of spatial learning; however, it is unclear whether this effect is related to the task-specific learning process or to nonspecific aspects of the testing procedure (e.g. exposure to the testing apparatus and exploration of the environment). Herein, we show that Arc-positive cells numbers are increased to the same extent in the granule cell layer after both acquisition of a single spatial learning event in the active place avoidance task and exploration of the testing environment, as compared to naïve (i.e. caged) mice. Repeated exposure the testing apparatus and environment did not reduce Arc expression. Furthermore, Arc expression did not correlate with performance in both adult and aged animals, suggesting that exploration of the testing environment, rather than the specific acquisition of the active place avoidance task, induces Arc expression in the dentate granule cell layer. These findings thus suggest that Arc is an experience-induced immediate-early gene.


Ocular artifacts in EEG and event-related potentials. I: Scalp topography.
The ocular artifacts that contaminate the EEG derive from the potential difference between the cornea and the fundus of the eye. This corneofundal or corneoretinal potential can be considered as an equivalent dipole with its positive pole directed toward the cornea. The cornea shows a steady DC potential of approximately +13 mV relative to the forehead. Blink potentials are caused by the eyelids sliding down over the positively charged cornea. The artifacts from eye-movements result from changes in orientation of the corneo-fundal potential. The scalp-distribution of the ocular artifacts can be described in terms of propagation factors--the fraction of the EOG signal at periocular electrodes that is recorded at a particular scalp location. These factors vary with the location of the scalp electrode. Propagation factors for blinks and upward eye-movements are significantly different.


Social Box: Influence of a New Housing System on the Social Interactions of Stallions When Driven in Pairs.
In order to improve the housing conditions of stallions in individual boxes, we tested a so-called "social box" allowing increased physical contact between neighbouring horses. This study investigated whether housing stallions in social boxes changes the number of social interactions during carriage driving. We hypothesised that the stay in social boxes would decrease the number of unwanted social interactions between stallions when driven in pairs. Eight Franches-Montagnes breeding stallions were observed when driven in pairs with a "neutral" stallion housed in a so-called "conventional box", strongly limiting physical contact. They were driven on a standardised route over the course of four days before, during, and after being housed in social boxes. The type and frequency of behaviours of the pairs and the interventions of the groom and the driver during the test drives were assessed live and using video recordings. Results from linear mixed-effect models show that unwanted social interactions decreased during and after the stallions were housed in the social box (p < 0.001). Stallions' interactions also decreased over the four days (p < 0.01), suggesting a habituation to the test conditions by learning not to interact, or by subtly settling dominance. The social box tended to decrease unwanted social behaviours of stallions driven in pairs and could therefore be used as an environmental enrichment for horses.


Technology platform for facile handling of 3D hydrogel cell culture scaffolds.
Hydrogels are used extensively as cell-culture scaffolds for both 2D and 3D cell cultures due to their biocompatibility and the ease in which their mechanical and biological properties can be tailored to mimic natural tissue. The challenge when working with hydrogel-based scaffolds is in their handling, as hydrogels that mimic e.g. brain tissue, are both fragile and brittle when prepared as thin (sub-mm) membranes. Here, we describe a method for facile handling of thin hydrogel cell culture scaffolds by molding them onto a polycaprolactone (PCL) mesh support attached to a commonly used Transwell set-up in which the original membrane has been removed. In addition to demonstrating the assembly of this set-up, we also show some applications for this type of biological membrane. A polyethylene glycol (PEG)-gelatin hydrogel supports cell adhesion, and the structures can be used for biological barrier models comprising either one or multiple hydrogel layers. Here, we demonstrate the formation of a tight layer of an epithelial cell model comprising MDCK cells cultured over 9 days by following the build-up of the transepithelial electrical resistances. Second, by integrating a pure PEG hydrogel into the PCL mesh, significant swelling is induced, which leads to the formation of a non-adherent biological scaffold with a large curvature that is useful for spheroid formation. In conclusion, we demonstrate the development of a handling platform for hydrogel cell culture scaffolds for easy integration with conventional measurement techniques and miniaturized organs-on-chip systems.


Anticardiolipin antibodies in Behçet's syndrome: a predictor of a more severe disease.
A high incidence of anticardiolipin antibodies were detected in 7 of 20 patients (35%) with Behçet's Syndrome. Three patients had IgG-ab, three had IgM-ab and one had both IgG and IgM antibodies. IgG-ACA was detected mainly in patients with ocular disease (30%) and one of them also has cerebral vascular disease. A lower incidence of ACA was found in the patients taking steroids compared with the ones taking other drugs. This work draws attention to the more severe disease present in patients with ACA and also the possibility of such tests become negative in patients taking immunosuppressive drugs.


How psychosis interrupts the lives of women and men differently: a qualitative comparison.
Psychosis disrupts how persons experience themselves and their lives. Despite knowledge that gender differences have been noted in presentation and course of psychosis symptoms, little is known about differences in how men and women experience these disruptions. The narratives of 26 men and 27 women diagnosed with psychosis, matched on age, education, and race, of the impact of psychosis on one's life, were compared. Using secondary data from semi-structured interviews, themes were identified using inductive, phenomenological qualitative analyses. Women and men discussed psychosis-related interruptions to the roles and relationships that shape their identity. Both genders discussed the impact of psychosis on their sense of self, work, and relationships. Nuanced gender differences emerged, informing unique areas of challenge related to (1) parenting and loss of parenting roles, (2) work and loss or changes in work trajectories, (3) isolation and strain on interpersonal relationships, and (4) manifestations of stigma. Psychosis may disrupt distinct aspects of life for men and women. Each gender faces the intersection of socially informed expectations that impact one's experiences of stigma, expectations of others, and manifestations of losses felt in one's role and sense of identity. Findings inform important considerations for therapy and other services. Gender-based socialized expectations, losses, and challenges that accompany psychosis are important areas for therapeutic consideration. Current treatments may neglect challenges that are more commonly experienced by women with psychosis.


Intestinal growth factors: potential use in the treatment of inflammatory bowel disease and their role in mucosal healing.
A key feature of inflammatory bowel disease (IBD) is impaired epithelial repair. Human growth factors comprise an array of signaling molecules that lead to ligand-specific signal transduction. Their downstream effects are associated with several cellular functions including epithelial healing in response to injury. Several studies have described specific growth factor deficiencies in patients with IBD, implicating their role in disease pathophysiology. The aim of this review was to describe currently known enterocyte-targeted growth factors, their mechanisms of action, and their potential therapeutic utility. The National Library of Medicine (http://www.pubmed.gov) and meeting abstracts were searched using the following terms: growth factor, intestine, colon, inflammatory bowel disease, Crohn's disease, ulcerative colitis, colitis, animal model, transforming growth factor, bone morphogenetic protein, activins, growth hormone, fibroblast growth factor, epidermal growth factor (EGF), keratinocyte growth factor (KGF), glucagon-like peptide II, granulocyte macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), vascular endothelial growth factor (VEGF) inhibitors, and trefoil factors. Several growth factors are therapeutic candidates in IBD. Growth hormone, KGF, EGF, teduglutide, GM-CSF/G-CSF have entered early clinical trials, whereas others are currently in preclinical evaluation. There are several growth factors responsible for epithelial repair. Preliminary studies using recombinant growth factors seem promising in IBD preclinical and clinical trials.


Three-point association of HLA-A,B,Bf haplotypes deduced in 200 parents of 100 families.
The association of HLA-A and -B antigens with Bf alleles was investigated in 200 parents from 100 unrelated families. There were significant associations between HLA-A3 and Bf-F, B7 and Bf-S, B8 and Bf-S, B12 and Bf-F, and BW 35 and Bf-F. Three-point HLA-A,B,Bf haplotype frequencies, linkage disequilibrium parameters, and chi-square values were determined both from the genotype and from the phenotype data. Although the HLA-B,Bf associations involve antigens that are also present in the highly associated A,B and B,D haplotypes of the Caucasian population, there was--with the possible exception of HLA-A3,B7,Bf-S--no significant three-point association for HLA-A,B,Bf.


A validated prognostic nomogram for patients with H3 K27M-mutant diffuse midline glioma.
H3 K27M-mutant diffuse midline glioma (H3 K27M-mt DMG) is a rare, highly invasive tumor with a poor prognosis. The prognostic factors of H3 K27M-mt DMG have not been fully identified, and there is no clinical prediction model for it. This study aimed to develop and validate a prognostic model for predicting the probability of survival in patients with H3 K27M-mt DMG. Patients diagnosed with H3 K27M-mt DMG in the West China Hospital from January 2016 to August 2021 were included. Cox proportional hazard regression was used for survival assessment, with adjustment for known prognostic factors. The final model was established using the patient data of our center as the training cohort and data from other centers for external independent verification. One hundred and five patients were ultimately included in the training cohort, and 43 cases from another institution were used as the validation cohort. The factors influencing survival probability in the prediction model included age, preoperative KPS score, radiotherapy and Ki-67 expression level. The adjusted consistency indices of the Cox regression model in internal bootstrap validation at 6, 12, and 18 months were 0.776, 0.766, and 0.764, respectively. The calibration chart showed high consistency between the predicted and observed results. The discrimination in external verification was 0.785, and the calibration curve showed good calibration ability. We identified the risk factors that affect the prognosis of H3 K27M-mt DMG patients and then established and validated a diagnostic model for predicting the survival probability of these patients.


[Von Willebrand factor].
During the last decade a considerable amount of research has been dedicated to von Willebrand factor. In the first part of this article recent data on primary structure of von Willebrand factor and biosynthesis of von Willebrand factor multimers are presented. The second part focuses on the role of von Willebrand factor in hemostasis.


Comparative study on the pollution status of organochlorine pesticides (OCPs) and bacterial community diversity and structure between plastic shed and open-field soils from northern China.
The pollution status of organochlorine pesticides (OCPs) and microbial community in plastic shed and open-field soils may be different due to the significant variations in environmental factors between the two cultivation modes. However, the differences remain unclear. We conducted a regional-scale survey to investigate the pollution level, distribution, and sources of 20 OCPs, and to evaluate the soil physicochemical properties and bacterial community in soils from plastic shed and open-field locating the north areas of China. We found that levels of total OCPs in the plastic shed soils were significantly higher than those in the nearby open-field soils. Most of these OCPs were attributed to historical application, except for dichlorodiphenyltrichloroethanes (DDTs) due to the fresh input along with dicofol application. Soil pH (for both cultivation modes) and total organic carbon (TOC) content (only for plastic sheds) were significantly correlated with the total OCP concentrations. Additionally, microbial diversity and richness were generally lower in plastic shed soils than in nearby open-field soils for each region. The bacterial community variation among different regions might be principally determined by the soil type. Soil pH had the greatest impact on the microbial community across all plastic shed and open-field samples. These results provide a better understanding of the environmental impact and ecological risk of OCPs in soils with different cultivation modes.


Proteomic analysis of the mouse brain after repetitive exposure to hypoxia.
Hypoxic preconditioning (HPC) is known to have a protective effect against hypoxic damage; however, the precise mechanisms involved remain unknown. In this study, an acute and repetitive hypoxia mouse model, two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) coupled with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/TOF-MS), and Western blot experiments were used to identify the differential expression of key proteins in the mouse brain during HPC. Approximately 2100 2D-DIGE spots were observed following gel imaging and spot detection. Significant differences (p < 0.05) in the expression of 66 proteins were observed between the 3× HPC treatment group and the control group, 45 proteins were observed between the 6× HPC treatment group and the control group, and 70 proteins were observed between the 3× HPC treatment group and the 6× HPC group. Consistent results among Western blot, 2D-DIGE and MS methods were observed for the proteins, ATP synthase subunit alpha, malate dehydrogenase, guanine nucleotide-binding protein subunit beta-1 and proteasome subunit alpha type-2. The proteins associated with ATP synthesis and the citric acid cycle were down-regulated, while those linked to glycolysis and oxygen-binding were up-regulated. This proteomic analysis of the mouse brain after HPC furthers understanding of the molecular pathways involved in the protective effect of HPC and these findings provide new insight into the mechanisms of hypoxia and HPC.


Encapsulation of isoflavone with milk, maltodextrin and gum acacia improves its stability.
This study was carried out for extraction of soy isoflavones and entrapment of the isoflavones so obtained into whole milk via encapsulation techniques. Three different solvent (ethanol, methanol and acetonitrile) were used for the extraction of isoflavone using three stage of extraction. The extracted isoflavone was encapsulated into 200 ml of whole milk by spray drying using different concentrations of gum acacia (4, 6 and 8% w/v) and 10% w/v maltodextrin DE 18. The ratio between cores to coating materials was 1:10. Though acetonitrile extracted higher amount of isoflavone, ethanol was selected for subsequent studies of extraction of isoflavone, as per the legislations regarding use of Food-grade solvents. There was no significant difference (p > 0.5) among all three samples 4% gum acacia+10% maltodextrin (A), 6% gum acacia+10% maltodextrin (B) and 8% gum acacia+10% maltodextrin (C) in terms of moisture content and hygroscopicity. However, insolubility index showed that sample A possessed a higher solubility index. Encapsulation techniques suggested that sample A showed higher encapsulation efficiency than others. Statistical analysis suggested that there was no significant difference among samples A, B and C during storage at 4°C for the time period (30 days) studied, in terms of isoflavone retention rate. However, samples stored at 25 and 37°C showed significant difference in the retention rate. Among all the three samples, sample B showed significantly lower isoflavone degradation rate of 3.80, 4.07 and 4.70 × 10-3/day at 4, 25 and 37°C, respectively. The highest amount of isoflavone degradation was observed at 37°C. Results indicate that isoflavone can be encapsulated using a combination of gum acacia either 4 or 6% w/v and 10% maltodextrin along with milk proteins at 4°C for longer shelf life.


Non-invasive aortoiliac assessment.
To assess the accuracy of Duplex ultrasound in the assessment of aortoiliac disease. Prospective, semi-blind study. Vascular laboratory and radiology departments, University Hospital. Ninety-two patients underwent assessment of the aortoiliac segment by femoral pulse palpation, Duplex ultrasound and biplanar arteriography. Of these 184 aortoiliac segments, 68 were also assessed by intraarterial pressure measurements and 80 by magnetic resonance angiography (MRA). Femoral pulses were abnormal in all 32 occluded aortoiliac segments. Of 152 patent segments, femoral pulse palpation was misleading in 50 (33%). MRA detected all occlusions and had a sensitivity of 71% and specificity of 68% for stenoses, compared to arteriography. Colour flow Duplex misdiagnosed four occlusions as stenoses. Duplex had a sensitivity of 91% and specificity of 93% for stenoses when compared to arteriography. Two stenoses, detected by Duplex and confirmed by pressure gradients, were missed by arteriography. Pressure measurements remain the gold standard for aortoiliac examination, arteriography providing only morphological information. The limitations of femoral pulse palpation should be appreciated. Although MRA was faster, Duplex examination proved slightly more sensitive to stenoses. At present, colour Duplex provides the best non-invasive assessment of aortoiliac disease and could prevent unnecessary arteriograms.


Caspase-cleaved tau is senescence-associated and induces a toxic gain of function by putting a brake on axonal transport.
The microtubule-associated protein tau plays a central role in tauopathies such as Alzheimer's disease (AD). The exact molecular mechanisms underlying tau toxicity are unclear, but aging is irrefutably the biggest risk factor. This raises the question of how cellular senescence affects the function of tau as a microtubule regulator. Here we report that the proportion of tau that is proteolytically cleaved at the caspase-3 site (TauC3) doubles in the hippocampus of senescent mice. TauC3 is also elevated in AD patients. Through quantitative live-cell imaging, we show that TauC3 has a drastically reduced dynamics of its microtubule interaction. Single-molecule tracking of tau confirmed that TauC3 has a longer residence time on axonal microtubules. The reduced dynamics of the TauC3-microtubule interaction correlated with a decreased transport of mitochondria, a reduced processivity of APP-vesicle transport and an induction of region-specific dendritic atrophy in CA1 neurons of the hippocampus. The microtubule-targeting drug Epothilone D normalized the interaction of TauC3 with microtubules and modulated the transport of APP-vesicles dependent on the presence of overexpressed human tau. The results indicate a novel toxic gain of function, in which a post-translational modification of tau changes the dynamics of the tau-microtubule interaction and thus leads to axonal transport defects and neuronal degeneration. The data also introduce microtubule-targeting drugs as pharmacological modifiers of the tau-microtubule interaction with the potential to restore the physiological interaction of pathologically altered tau with microtubules.


Validation of phlebotomy performance metrics developed as part of a proficiency-based progression initiative to mitigate wrong blood in tube.
The purpose of this study was to (1) characterise the procedure of phlebotomy, deconstruct it into its constituent parts and develop a performance metric for the purpose of training healthcare professionals in a large teaching hospital and to (2) evaluate the construct validity of the phlebotomy metric and establish a proficiency benchmark. By engaging with a multidisciplinary team with a wide range of experience of preanalytical errors in phlebotomy and observing video recordings of the procedure performed in the actual working environment, we defined a performance metric. This was brought to a modified Delphi meeting, where consensus was reached by an expert panel. To demonstrate construct validity, we used the metric to objectively assess the performance of novices and expert practitioners. A phlebotomy metric consisting of 11 phases and 77 steps was developed. The mean inter-rater reliability was 0.91 (min 0.83, max 0.95). The expert group completed more steps of the procedure (72 vs 69), made fewer errors (19 vs 13, p=0.014) and fewer critical errors (1 Vs 4, p=0.002) than the novice group. The metrics demonstrated construct validity and the proficiency benchmark was established with a minimum observation of 69 steps, with no critical errors and no more than 13 errors in total.


Three cases of amyotrophic lateral sclerosis in a common occupational environment.
Three unrelated school teachers taught in the same school classroom for 2-5 years and subsequently developed amyotrophic lateral sclerosis (ALS) over an 18-year period. This clustering was not accompanied by an increased death rate for ALS in the county where the teachers lived and worked. Statistical analysis revealed that ALS as the cause of death for three teachers from the same school would be highly improbable as a random event. These findings suggest that the patient's shared school environment may have been a source of exposure to an agent pathogenetically significant in the development of their disease.


Predictors of the variation of the ejection fraction during exercise in patients with and without coronary heart disease.
The aim of this study was to determine the factors influencing the different response of the ejection fraction (EF) of the left ventricle at exercise observed in patients with and without significant coronary heart disease. We have studied 98 patients referred for coronary angiography (82 men, 16 women), of whom 49 patients had a previous myocardial infarction and 71 patients had significant coronary heart disease. Exercise testing was performed and combined with a cardiac blood pool imaging at equilibrium. The variation of the EF between rest and peak exercise (delta EF) was measured. Twelve clinical, exercise-related, isotopic, and coronary arteriographic variables were examined in a linear univariate and statistical analysis. In the univariate regression, seven variables were significant regressors on the delta EF. In the multivariate regression, only four variables were significant regressors on the delta EF. Three independent predictors were found: the rate-pressure product, the ST depression, and the occurrence of a previous myocardial infarction. These three independent predictors reflect the myocardial functional reserve.


Spinal metastases from myxoid liposarcoma warrant screening with magnetic resonance imaging.
Myxoid liposarcoma (MLS) has an unusual tendency for extrapulmonary metastasis, particularly to the spine and soft tissues. The objective of this study was to determine the prevalence of spinal metastasis, treatment outcomes, and optimal screening method for spinal metastasis in patients with MLS. Data from patients with had spinal metastases were obtained from the authors' institutional soft tissue sarcoma database. The accuracy with which positron emission tomography (PET) scans and bone scans identified metastatic lesions was compared with the accuracy of magnetic resonance imaging (MRI). Clinical response to treatment was based on pain, neurologic scores, and survivorship analysis. There were 33 patients who developed spinal metastasis after a median 36 months of follow-up (range, from 7.5 months to 33 years). Known spinal metastases were detected by bone scans in 16% of patients and by PET scans in 14% of patients. Patients who underwent surgery had high-grade spinal cord compression more often than patients who did not undergo surgery (72% vs 19%, respectively; P = .002). Pain and neurologic function were improved or maintained in all patients who received radiation alone (n = 8 patients) and in all but 1 patient who underwent surgery (n = 18 patients). The median overall survival was 51.4 months from the time of primary diagnosis and 21.9 months from the time of first metastasis. Bone scans and PET scan lack sufficient sensitivity to detect spinal metastasis from MLS. Treatment of metastasis is palliative, but local treatment can yield long-term disease control in select patients. Screening with whole-spine MRI may lead to the earlier detection of spinal metastasis.


Zn2+ inhibits spatial memory and hippocampal place cell representation through high-affinity binding to the NMDA receptor GluN2A subunit.
A subset of glutamatergic neurons in the forebrain uses labile Zn2+ as a co-transmitter alongside glutamate. Synaptic Zn2+ plays a key role in learning and memory processes, but its mechanisms of action remain poorly understood. Here, we used a knock-in (KI) mouse line carrying a point mutation at the GluN2A Zn2+ binding site that selectively eliminates zinc inhibition of NMDA receptors. Ablation of Zn2+-GluN2A binding improves spatial memory retention and contextual fear memory formation. Electrophysiological recording of hippocampal neurons in the CA1 area revealed a greater proportion of place cells and substantial place field remapping in KI mice compared to wildtype littermates. Persistent place cell remapping was also seen in KI mice upon repeated testing suggesting an enhanced ability to maintain a distinct representation across multiple overlapping experiences. Together, these findings reveal an original molecular mechanism through which synaptic Zn2+ negatively modulates spatial cognition by dampening GluN2A-containing NMDA receptor signaling.


Parenting: what are the critical attributes?
Identifying the critical attributes of the parenting concept. The parenting attributes are the process, activity, and interaction regarding to rearing and educating a child, which is undertaken by parent or parental figure. Those aim at developing a child's growth, development, and well-being. The specific parenting attributes can be applied to practice, education, and research in the area of pediatric, family and community health care.


Identification of the receptor for toxin II alpha from scorpion Leirus quinquestriatus in isolated lobster and squid nerve membranes.
1. Isolated membranes from lobster and squid nerves were labeled with Leirus quinquestriatus toxin II alpha. 2. Dinitrofluorophenylazide 125I-ScTx derivative was synthesized and covalently bound to the membranes. 3. It was found that the membranes of these nonmyelinated nerves present a single polypeptide located at approximately 220-230 Kd range labeled by the ScTx. 4. Rat brain membrane used for comparison presents two polypeptidic zones at 220 and 50 Kd labeled by the toxin.


A patient with fulminant systemic vasculitis type polyarteritis nodosa and negative histology of small bowel infarction.
A patient with fatal polyarteritis nodosa is described. In spite of overwhelming clinical signs the diagnosis of PAN could not be confirmed by angiography nor histology of macroscopic clearly involved small bowel tissue. Histology of lower limb tissue after amputation confirmed the diagnosis.


Power-free colorimetric biosensing of foodborne bacteria in centrifugal tube.
Early finding of pathogens is significant to avoid foodborne diseases. Here, a novel lab-in-centrifugal-tube colorimetric biosensor was reported for Salmonella typhimurium detection using immune nickel nanowires (NNWs) to form capture nets for specific bacterial separation, gold@platinum nanozymes (GPNs) to mark target bacteria for effective signal amplification, and a smartphone App to analyze color change for quantitative bacterial determination. A 3D-printed cylindrical magnetic separator with air pressure self-regulating structure and NNW capture nets was elaboratively constructed and assembled inside the disposable centrifuge tube to simply perform the bacterial separation, label, wash, coloration and detection. Under optimal conditions, Salmonella typhimurium could be quantitatively detected in 2 h with a low detection limit of 21 CFU/mL. The recovery of target bacteria in spiked pork samples ranged from 87.0% to 97.6% with the averaged recovery of 93.9%. This biosensor was Affordable, Sensitive, Specific, User-friendly, Rapid and robust, Equipment-free and Deliverable to end-users (ASSURED), and had shown the potential for point-of-care testing of foodborne pathogens to ensure food safety.


Microbial community succession during crude oil-degrading bacterial enrichment cultivation and construction of a degrading consortium.
Microbial community succession during the enrichment of crude-oil-degrading bacteria was analyzed using Illumina high-throughput sequencing to guide bacterial isolation and construction of a bacterial consortium. Community change occurred in 6 days; the most abundant phylum changed from Proteobacteria to Actinobacteria; the most abundant genera were Dietzia and unspecified_Idiomarinaceae. Two crude oil-degrading strains, Rhodococcus sp. OS62-1 and Dietzia sp. OS33, and one weak-crude-oil-degrading strain, Pseudomonas sp. P35, were isolated. A consortium comprising Rhodococcus sp. OS62-1 and Pseudomonas sp. P35 showed the highest crude-oil-degrading efficiency, reaching 85.72 ± 3.21% within 7 days, over a wide pH range (5-11) and salinity (0-80 g·L-1). Consumption of saturated hydrocarbons, aromatic hydrocarbons, and resins was greater by the consortium than by a single strain, as was degradation of short-chain-alkanes (C13-C17) according to gas-chromatography. The bacterial consortium provides technical support for bioremediation of crude oil pollution.


Detection of Helicobacter pylori gene by means of immunomagnetic separation-based polymerase chain reaction in feces.
Detection of Helicobacter pylori is usually performed by culture, polymerase chain reaction (PCR), histology, or urease test on gastric biopsy samples. Although methods based on feces are non-invasive, their sensitivity has been relatively low. In this study, to improve its sensitivity, immunomagnetic separation (IMS) was used as a pre-PCR step for direct detection of H. pylori in feces. Fresh fecal samples were taken from 72 patients attending for endoscopy. Of these, 57 patients had a positive H. pylori status according to the results of culture, histology, and PCR on gastric biopsy samples. Anti-H. pylori antibody-sensitized immunomagnetic beads were used to concentrate the bacteria. PCR was then performed to detect the H. pylori urease A-encoding gene. Of the 57 H. pylori-positive patients, 35 (61.4%) had positive fecal samples by IMS-based PCR method. None of the 15 H. pylori-negative patients had positive fecal samples. The sensitivity of this method was 61.4%, and the specificity 100.0%. This study confirms that non-invasive diagnosis of H. pylori infection could be made from feces by using IMS-based PCR.


A high-fat diet and multiple administration of carbon tetrachloride induces liver injury and pathological features associated with non-alcoholic steatohepatitis in mice.
The aim of the present study was to establish a progressive steatohepatitis mouse model because few reported animal models of non-alcoholic steatohepatitis (NASH) show the progression from fatty liver to steatohepatitis. C57BL/6N mice were fed a high-fat diet (HFD) to develop obesity and were either administered carbon tetrachloride (CCl4 ) eight times (0.05 mL/kg, s.c., once, followed by 0.1 mL/kg, s.c., seven times) or not. Serum parameters and hepatic histopathology were examined. In a separate experiment, CCl4 was administered subcutaneously from 0 to eight times to HFD-fed obese mice to investigate progressive changes. Markers of oxidative stress, inflammation and apoptosis, as well as histopathological changes in the liver, were analysed. The HFD-fed obese mice showed fatty liver but not steatohepatitis. In contrast, HFD-fed mice administered CCl4 eight times showed histopathological features of steatohepatitis (fatty liver, inflammation, hepatocellular ballooning and fibrosis) and increased serum alanine aminotransferase levels. However, the multiple administration of CCl4 to obese mice reduced the ratio of reduced glutathione to oxidized glutathione, superoxide dismutase activity and mitochondrial DNA copy number, leading to the development of chronic oxidative stress, increased numbers of apoptotic cells and increased levels of both tumour necrosis factor-α and transforming growth factor-β mRNA. The resulting inflammation led to increased hydroxyproline content in the liver and fibrosis. The present study demonstrates that multiple administration of CCl4 to HFD-fed obese mice induces chronic oxidative stress that triggers inflammation and apoptosis and leads to the development of fibrosis in the liver, resulting in progression from fatty liver to steatohepatitis. This murine model will be useful in the research of hepatic disorders.


Touching The Human Neuron: User-Centric Augmented Reality Viewing and Interaction of in-vivo Cellular Confocal Laser Scanning Microscopy (CLSM) Utilizing High Resolution zStack Data Sets for Applications in Medical Education and Clinical Medicine Using GLASS and Motion Tracking Technology.
This is the first report of a prototype that allows for real-time interaction with high-resolution cellular modules using GLASS© technology. The prototype was developed* using zStack data sets which allow for real-time interaction of low polygon and direct surface models exported from primary source imaging. This paper also discusses potential educational and clinical applications of a wearable, interactive, user-centric, augmented reality visualization of cellular structures. * Prototype was developed using IMARIS© , ImageJ© , MAYA® , 3DCOAT© , UNITY© , ZBrush© , OcculusRift® , LeapMotion© , and output for multiplatform function with Google GLASS© technology as its viewing modality.


[Simultaneous urodynamic and electromyographic micturition studies].
The clinical significance of micturition investigations is to be seen not only in the quantification of urethral obstructions e.g. after operations for stress incontinence. Such measurements in fact give information on very different forms of voiding dysfunctions with and without morphological substrates. Simultaneous EMG measurements permit further differentiation, especially in the case of purely functional neurogenic or mitigated neurogenic voiding dysfunctions.


Symptomatic and silent cerebral infarction following surgical clipping of unruptured intracranial aneurysms: incidence, risk factors, and clinical outcome.
Cerebral infarction (CI) associated with clipping of unruptured intracranial aneurysms (UIAs) has not been completely studied. The role of individual and operative characteristics is not known, and the risk of silent CI has not been well described. To determine the incidence, risk factors, and clinical outcome of postoperative CI, we retrospectively analyzed 388 consecutive patients undergoing clipping of UIAs between January 2012 and December 2015. We reviewed the pre- and postoperative computed tomography (CT) images of each patient. Postoperative CI was defined as a new parenchymal hypodensity in the vascular territory of treated artery. Patient-specific, aneurysm-specific, and operative variables were analyzed as potential risk factors. Functional outcome at discharge was assessed with the modified Rankin Scale (mRS). Postoperative CI was found in 49 (12.6%) patients, 29 of whom manifested neurological deficits. The incidences of symptomatic stroke and silent CI were 7.5 and 5.2%, respectively. Multivariate analysis showed that larger aneurysm size and history of hypertension were significantly associated with CI. Disability (mRS > 2) rate was 42.9% among patients with CI, which was substantially higher than that among patients without (0.9%). In conclusion, the incidence of CI following clipping of UIAs was not low. Larger aneurysm size and history of hypertension were independent risk factors. Postoperative symptomatic stroke correlated with an extremely high risk of disability. Silent CI was seemingly nondisabling, but the possible cognitive consequence is pending.


Evaluation of BEH C18, BEH HILIC, and HSS T3 (C18) column chemistries for the UPLC-MS-MS analysis of glutathione, glutathione disulfide, and ophthalmic acid in mouse liver and human plasma.
Glutathione (GSH), glutathione disulfide (GSSG), and ophthalmic acid (OA) are important biological oxidative stress biomarkers to be monitored in pathological and toxicological studies. With the advent of liquid chromatography tandem mass spectrometry (LC-MS-MS) technology, sensitive and selective analysis of these biomarkers in biological samples is now being performed routinely. Due to the hydrophilic and polar natures of GSH and its endogenous derivatives, achieving good retention, resolution, and peak shape is often a chromatographic challenge. In this study, three ultra-performance (UP) LC column chemistries (namely, BEH C18, BEH HILIC, and HSS T3 [C18]) are evaluated for the UPLC-MS-MS analysis of GSH, GSSG, and OA extracted from mouse liver and human plasma samples. The chromatographic parameters evaluated are retentivity, tailing factor, MS sensitivity, and resolution of the three analytes. Based on the optimized method for each column chemistry, our results indicate that the HSS T3 (C18) column chemistry affords the best retention and separation of these analytes when operated under the ultra high-pressure chromatographic conditions.


Activation of farnesoid X receptor (FXR) induces crystallin zeta expression in mouse medullary collecting duct cells.
Crystallin zeta (CRYZ) is a phylogenetically restricted water-soluble protein and provides cytoprotection against oxidative stress via multiple mechanisms. Increasing evidence suggests that CRYZ is high abundantly expressed in the kidney where it acts as a transacting factor in increasing glutaminolysis and the Na+/K+/2Cl- cotransporter (BSC1/NKCC2) expression to help maintain acid-base balance and medullary hyperosmotic gradient. However, the mechanism by which CRYZ is regulated in the kidney remains largely uncharacterized. Here, we show that CRYZ is a direct target of farnesoid X receptor (FXR), a nuclear receptor important for renal physiology. We found that CRYZ was ubiquitously expressed in mouse kidney and constitutively expressed in the cytoplasm of medullary collecting duct cells (MCDs). In primary cultured mouse MCDs, CRYZ expression was significantly upregulated by the activation and overexpression of FXR. FXR-induced CRYZ expression was almost completely abolished in the MCD cells with siRNA-mediated FXR knockdown. Consistently, treatment with FXR agonists failed to induce CRYZ expression in the MCDs isolated from mice with global and collecting duct-specific FXR deficiency. We identified a putative FXR response element (FXRE) on the CRYZ gene promoter. The luciferase reporter and ChIP assays revealed that FXR can bind directly to the FXRE site, which was further markedly enhanced by FXR activation. Furthermore, we found CRYZ overexpression in MCDs significantly attenuated hypertonicity-induced cell death possibly via increasing Bcl-2 expression. Collectively, our findings demonstrate that CRYZ is constitutively expressed in renal medullary collecting duct cells, where it is transcriptionally controlled by FXR. Given a critical role of FXR in MCDs, CRYZ may be responsible for protective effect of FXR on the survival of MCDs under hypertonic condition during dehydration.


Beneficial effects of Wuzhi Capsule on tacrolimus blood concentrations in liver transplant patients with different donor-recipient CYP3A5 genotypes.
Tacrolimus (Tac) is an immunosuppressant that is widely used to prevent allograft rejection in patients after liver transplantation. Its metabolism mainly depends on the cytochrome P450 3A5 (CYP3A5), which has genetic polymorphisms. Recently, a Chinese herbal medicine known as Wuzhi Capsule (WZC) was shown to increase Tac blood concentrations by inhibiting the activity of CYP3A in animal studies in rats. To date, it remains unexplored whether WZC can be efficiently used to enhance the blood concentration of Tac in liver transplant patients with different donor-recipient CYP3A5 genotypes. A total of 185 liver transplant patients were enrolled and two-way ANOVA was carried out, then they were divided into four groups according to the combinations of donor-recipient CYP3A5 phenotypes. WZC was given to patients when the dose of Tac was ≥4 mg, and the dose-adjusted C0 (C0 /D) of Tac measured twice in succession was ≤1 ng/ml/mg. The blood trough concentration of Tac (C0 ), C0 /D, and dose- and body weight-adjusted C0 (C0 /D/W) was analysed on days 7 and 14 after liver transplantation. The genotypes of donor and recipient or WZC had significant effects on C0, C0/D and C0/D/W. There were significant differences in the Tac blood concentrations between the groups. The recipient expression (*1)/donor expression (*1) (R+/D+) group had the lowest C0 , C0 /D and C0 /D/W among the four groups. Furthermore, a larger proportion of patients in the CYP3A5 expression groups required Tac dose adjustment to achieve a therapeutic effect and were given Tac with WZC. Notably, the use of WZC significantly increased the blood concentrations of Tac in the CYP3A5 expression groups and greater increases in the C0 /D and C0 /D/W were significantly associated with higher doses of WZC in the CYP3A5 expression groups. What is more, WZC reduced the hospitalization cost of patients to a certain extent. WZC significantly increased the C0 , C0 /D and C0 /D/W in the CYP3A5 expression groups and reduced the hospitalization expenses of patients to a certain extent. What is more, greater increases in the C0 /D and C0 /D/W were significantly associated with higher doses of WZC.


Prevalence of apical periodontitis in endodontically-treated maxillary and mandibular posterior teeth in a Saudi Arabian population: a cone-beam computed tomography study.
The aim of this retrospective study was to assess the prevalence of apical periodontitis (AP) in root canal-treated maxillary and mandibular posterior teeth in a Saudi Arabian population based on findings from images taken using cone-beam computed tomography (CBCT). The sample included 300 CBCT images from patients (150 females and 150 males) aged between 18 and 80 years old and they were analyzed to detect AP in endodontically treated maxillary and mandibular premolars and molars. Also, the correlation between the prevalence of AP and gender along with location (right/left side of jaw) were analyzed. Periapical lesions were defined as lamina dura gaps appearing disrupted around the width of periodontal ligament and apex at the apical third of the roots. There was higher prevalence of AP in maxillary (19.1%) when compared to mandibular posterior teeth (17.3%). Mandibular first and maxillary second molars were the most affected teeth with AP (P = 0.05 and P = 0.04, respectively). Also, there was higher prevalence of AP cases with a predilection toward males (Maxillary P = 0.005; Mandibular P = 0.00) and on the right side of the mandibular jaw (P = 0.03). In this study, the prevalence of AP had a significant association between gender with a predilection in males and location with a tendency to occur on the mandibular right side as observed in endodontically treated maxillary and mandibular posterior teeth using CBCT.


The concentration and chemical speciation of arsenic in the Nanpan River, the upstream of the Pearl River, China.
The concentration and chemical speciation of arsenic (As) in different environmental matrixes (water, sediment, agricultural soils, and non-agricultural soils) were investigated in the Nanpan River area, the upstream of Pearl River, China. The results did not show any obvious transport of As along the flow direction of the river (from upstream to downstream). Total As concentrations in sediment were significantly different from those in agricultural soil. According to the comparison to quality standards, the As in sediments of the studied area have potential ecological risks and a minority of the sampling sites of agricultural soils in the studied area were polluted with As. As speciations were analyzed using sequential extraction and the percentage of non-residual fraction in sediment predominated over residual fraction. We thus believe that As in the studied area was with low mobility and bioavailability in sediment, agricultural soils, and non-agricultural soils. However, the bioavailability and mobility of As in sediment were higher than in both agricultural and non-agricultural soils, and thus, special attention should be paid for the risk assessment of As in the river in future studies.


The effect of coffee consumption on the non-alcoholic fatty liver disease and liver fibrosis: A meta-analysis of 11 epidemiological studies.
Non-alcoholic fatty liver disease (NAFLD) is a widespread chronic liver disease. It is considered a multifactorial disorder that can progress to liver fibrosis and cause a worldwide public health concern. Coffee consumption may have a protective impact on NAFLD and liver fibrosis. However, the evidence from the previous studies is inconsistent. This meta-analysis summarizes available literature. This study comprises two meta-analyses. The first meta-analysis summarizes the effect of coffee consumption on NAFLD in those who did or did not drink coffee. The second analysis compares the risk of liver fibrosis development between NAFLD patients who did or did not drink coffee. Pooled risk ratios (RR) and confidence intervals (CI) of observational studies were estimated. Of the total collected 321 articles, 11 met our eligibility criteria to be included in the analysis. The risk of NAFLD among those who drank coffee compared to those who did not was significantly lower with a pooled RR value of 0.77 (95% CI 0.60-0.98). Moreover, we also found a significantly reduced risk of liver fibrosis in those who drink coffee than those who did not drink in the NAFLD patients with the relative risk (RR) of 0.68 (95% CI 0.68-0.79). Regular coffee consumption is significantly associated with a reduced risk of NAFLD. It is also significantly associated with decreased risk of liver fibrosis development in already diagnosed NAFLD patients. Although coffee consumption may be considered an essential preventive measure for NAFLD, this subject needs further epidemiological studies.


Knowledge and experiences with augmentative and alternative communication by paediatric nurses: a pilot study.
The current mixed methods research study was designed to describe the experience and opinions of paediatric acute care nurses at a single paediatric medical centre regarding augmentative and alternative communication (AAC). This research serves as the beginning of a line of research to determine needs that can be met through later education and training on AAC with paediatric acute care nurses. Mixed methods were used to gather survey and interview data from nurses at a single paediatric hospital to investigate their knowledge of AAC, perceived relevance of AAC for their patient populations, and preferences for training programmes. Quantitative survey and qualitative interview data indicated a need for AAC education for these paediatric nurses. Nurses demonstrated limited knowledge and experience with AAC but communicated both a desire and perceived need to know more to effectively care for their patients. Nurses reported limited AAC knowledge but expressed a desire for support to use AAC with their patients. AAC education and cross-sector collaboration with healthcare workers such as speech-language pathologists (SLPs) and acute care paediatric nurses are indicated to support effective communication when interacting with patients with complex communication needs (CCN).Implications for rehabilitationImproved background knowledge to develop and implement future nurse education on AAC.Foundation for cross-sector collaboration (e.g., nurses and SLPs) on AAC implementation in paediatric acute care settings.


Terminal proliferation and synaptogenesis following partial deafferentation: the reinnervation of the inner molecular layer of the dentate gyrus following removal of its commissural afferents.
The inner one-third of the dendritic region of the dentate gyrus granule cells in adult rats receives projections primarily from the commissural fibers of the contralateral hippocampus and the associational fibers of the ipsilateral hippocampus. At two to four days following the complete removal of the contralateral hippocampus, approximately 25% of the terminals in the inner molecular layer are observed degenerating. This provides an excellent model system to investigate possible terminal proliferation induced by deafferentation since (1) the experimental lesion is easily reproducible, (2) no retrograde reactions occur in the granule cells as a direct result of the lesion, (3) no shrinkage is detected in this region following commissural deafferentation, (4) the same dendritic region can be relocated precisely in each animal, and (5) the synaptic counts are highly consistent between animals. Results from this study and from previous investigations demonstrate that the commissural projection is contained within a 0-80 mu zone directly above the granule cell layer; Complete photomontages of this zone were taken, but only the 40-80 mu zone was quantified for neuronal and glial changes in three normal, five 2- to 4-day, and five 50- to 75-day postlesion animals. The average synaptic count dropped to 64% of control values by 2 to 4 days, returned to 97% by 50- to 75 days postlesion, The number of terminals showing multiple synaptic contacts increased slightly in the long-term animals. Measurements of average terminal area showed no change between the short- and long-term survival groups. These results indicate that this dendritic region is reinnervated following partial deafferentation and that the reinnervation is due primarily to the formation of new terminals rather than the expansion of pre-existing terminals.


Anti-Tumor Activity and Underlying Mechanism of Phomoxanthone B in MCF7 Cells.
Xanthones are a class of heterocyclic natural products, which are promising sources of anti-cancer leads. Phomoxanthone B (PXB) and Phomoxanthone A(PXA)are xanthone dimers. PXA is wellstudied as an anti-cancer agent, but PXB is not. In our study, PXB was isolated from the endophytic fungus Phomopsis sp. By254. The purpose of this study was to identify the underlying anti-tumor mechanisms of PXB in breast cancer MCF7 cell line. Apoptosis, cell cycle, proliferation, invasion, and migration assays were used to assess the anti-tumor activity of PXB. RNA sequencing was used to analyze the effect of PXB treatment on gene expression in MCF7 cells. PXB showed cytotoxicity towards a variety of tumor cells, especially MCF7 cells. PXB inhibited the migration and invasion, arrested cell cycle at G2/M phase, and induced apoptosis associated with caspase-3 activation in MCF7 cells. The detailed transcriptome analysis revealed that PXB affected several pathways related to tumorigenesis, metabolisms, and oxidative phosphorylation in MCF7 cells. KEGG transcriptome analysis revealed that PXB upregulated pro-survival signal pathways, such as MAPK, PI3K-AKT, and STAT3 pathways. We found that PXB also significantly upregulated the expression of IL24, DDIT3, and XAF1, which may contribute to PXB-induced apoptosis. We further found that PXB may downregulate oxidative phosphorylation by decreasing the expression of electron transport chain genes, especially MT-ND1, which is a potential unfavorable prognostic marker for ER-positive breast cancer. PXB exerts strong cytotoxicity against human tumor cells and has a potential for ER-positive breast cancer treatment.


Efficacy of a modified short fully covered self-expandable metal stent for perihilar benign biliary strictures.
Endoscopic management of a benign biliary stricture (BBS) on the hilum is complicated and challenging. Although the placement of a fully covered self-expandable metal stent (FCSEMS) is possible to increase effectiveness, stent migration and stent-induced adverse events are problematic. We aimed to evaluate the usefulness of a modified short FCSEMS with a long lasso in patients with a difficult perihilar BBS. Patients with perihilar BBS within 2 cm from the hilar confluence that failed initially with plastic stents were enrolled. A modified short FCSEMS was deployed and then removed 5-6 months later. The primary outcome was clinical success. Other technical success, adverse events, endoscopic success of stent removal, and recurrence of stricture during the follow-up period were measured. Endoscopic intraductal placement was technically successful in all patients (n = 19). Combined contralateral plastic stent placement was performed in 13 patients (68.4%). The median duration of stent placement was 163 days (range, 138-196 days). Endoscopic stent removal was successful in all patients except one spontaneous distal migration. Stricture resolution without de novo focal stricture occurrence was 100%. Endoscopic stone removal after stricture improvement was successful in all 13 patients with bile duct stones above the stricture. During a follow-up period (median 635 days) after stent removal, only one recurrence developed. Temporary placement of a modified intraductal short FCSEMS with or without a contralateral plastic stent improved perihilar BBS in patients that primarily failed by plastic stents. Combined biliary stones were also successfully removed after stricture resolution.


Diabetic foot disease and the risk of major clinical outcomes.
Data on the long-term consequences of diabetic foot disease (DFD) are scarce. We examined the association between DFD and major clinical outcomes in patients with diabetes in the general population. We conducted a prospective cohort analysis of 1,428 participants with diabetes in the Atherosclerosis Risk in Communities Study. DFD and four clinical outcomes (nontraumatic lower-extremity amputation, cardiovascular disease, major fall, and death) were captured through 2018 using administrative data. We used Cox regression models to evaluate the association between incident DFD (modeled as a time-varying exposure) and the subsequent risk of clinical outcomes. During over two decades of follow-up (1996-1998 to 2018), the cumulative incidence of DFD was 33.3%. Risk factors for DFD included older age, poor glycemic control, long diabetes duration, and prevalent vascular disease (chronic kidney disease, retinopathy, cardiovascular disease). Following incident DFD, the five-year cumulative incidence of major clinical outcomes was 38.9% for mortality, 25.2% for cardiovascular disease, 14.5% for nontraumatic lower-extremity amputation, and 13.2% for major fall. DFD remained associated with all four clinical outcomes after multivariable adjustment, with hazard ratios ranging from 1.5 (cardiovascular disease) to 34.7 (lower-extremity amputation). DFD is common and confers substantial risk for major morbidity and mortality.


Monthly prospective analysis of hematopoietic chimerism after allogeneic hematopoietic cell transplantation.
Hematopoietic chimerism (HpC) was assayed monthly using a sensitive, polymerase chain reaction (PCR) -based method in consecutive patients. Between January 1998 and April 2002, 181 patients underwent non-T cell depleted allogeneic hematopoietic cell transplantation (HCT). A total of 163 patients were evaluable for HpC at 1 month (11 early deaths; no informative band for HpC analysis/no genomic DNA in seven). In all, 53 of 163 patients (33%, median recipient DNA of 15% (range 5-95)), 39 of 151 patients (26%), and 27 of 142 patients (19%) showed mixed chimerism (MC) at 1, 2, and 3 months after HCT, respectively. Conditioning regimen (busulfan-fludarabine-ATG vs BuCy, relative risk 3.99 (95% CI 1.16-10.92)), neutrophil engraftment (>/=day 17 vs </=day 16, relative risk 2.49 (95% CI 1.14-5.41)), and acute graft-versus-host disease within 30 days (none vs yes, relative risk 4.78 (95% CI 1.50-15.17)) were independent variables that showed significant correlation with having >/=5% recipient DNA at 1 month. Five patients experienced secondary graft failure. All five patients showed MC at 1 month with median recipient DNA of 40%. None of the 109 patients with complete chimerism experienced graft failure (P=0.002). Our study showed that MC shown on monthly analysis of HpC after allogeneic HCT is a significant predictor of secondary graft failure. Bone Marrow Transplantation (2003) 32, 423-431. doi:10.1038/sj.bmt.1704147


Off-pump Coronary Artery Bypass Grafting in patients on chronic hemodialysis.
Off-pump Coronary Artery Bypass Grafting(CABG) has gained much support but has been scarcely reported in patients on chronic hemodialysis (HD). The details of CABG for such patients on chronic HD are presented. Between January 1998 and December 1999, off-pump CABG was performed in 5 patients on chronic HD. All patients presented with unstable angina pectoris. The indication for off-pump CABG was suitable coronary anatomy (N = 5) and expected high risks associated with cardiopulmonary bypass due to reoperation (N = 1) and impaired cerebral blood flow (N = 2). The approach was median sternotomy (N = 4) and left anterior short thoracotomy (N = 1). The mean graft number was 2.0 +/- 1.0 (range, 1-3). In situ arterial grafts were mainly used: the in situ left internal thoracic artery was used in four patients, the in situ right gastroepiploic artery in two, the in situ right internal thoracic artery in two, and the saphenous vein in two patients. There were no deaths or complications. Nine of 10 grafts were confirmed fully patent by postoperative angiography before hospital discharge. In a mean follow-up of 17.2 months, no patients had developed recurrence of angina or any cardiac symptom. Off-pump CABG using in situ arterial grafts can be performed on patients on chronic hemodialysis subject to suitable anatomy of the target coronary arteries.


Vibration, acoustic and emission characteristics of the chlorella vulgaris microalgae oil in compression ignition engine to mitigate environmental pollution.
The petroleum fuel demand with high price and its exhaustion imposes a pressure to find an alternative. The fossil fuel shortage has been deteriorating over the past few years, because of the rapid increase in population. Many attempts have been made to increase the quality of biofuel with additives. In this paper, two types of nanoparticles such as carbon nanotubes (CNT) and alumina (Al2O3) in chlorella microalgae biofuel were analyzed by experimental method. The added CNT and alumina act as a catalyst that induces complete combustion with retarded emissions. In addition to above, the noise and vibration qualities are also measured. A series of test conducted using single cylinders, four stroke, naturally aspirated compression ignition diesel engine was run by using pure diesel and also different fuel blends 'such as B10CNT50A50 Chlorella (Microalgae Biodiesel 10% + Diesel 90% + CNT 50 ppm), B20CNT50A50 (Microalgae Biodiesel 20% + Diesel 80% + CNT 50 ppm + Al2O3 50 ppm) and B30CNT50A50 (Microalgae Biodiesel 30% + Diesel 70% + CNT 50 ppm + Al2O3 50 ppm). At a constant load condition, all experimental tests were conducted at four different speeds such as 1500 rpm, 2000 rpm, 2500 rpm and 3000 rpm. The reference fuel of diesel B0 results was compared with blended fuel. From the results, it has been found that the nano additives of CNT and alumina reduced the greenhouse gas emissions of CO compared to plain diesel. Only considering the blended fuel, as the percentage of biofuel increases, the emission of nitric oxide and carbon dioxide is decreased with significant reduction in the amount of noise and vibration and also the combustion and performance qualities were also improved. The highest benefit in terms of all factors was achieved in the fuel blend of B30A50CNT50 amongst the other blends.


BlobFinder, a tool for fluorescence microscopy image cytometry.
Images can be acquired at high rates with modern fluorescence microscopy hardware, giving rise to a demand for high-speed analysis of image data. Digital image cytometry, i.e., automated measurements and extraction of quantitative data from images of cells, provides valuable information for many types of biomedical analysis. There exists a number of different image analysis software packages that can be programmed to perform a wide array of useful measurements. However, the multi-application capability often compromises the simplicity of the tool. Also, the gain in speed of analysis is often compromised by time spent learning complicated software. We provide a free software called BlobFinder that is intended for a limited type of application, making it easy to use, easy to learn and optimized for its particular task. BlobFinder can perform batch processing of image data and quantify as well as localize cells and point like source signals in fluorescence microscopy images, e.g., from FISH, in situ PLA and padlock probing, in a fast and easy way.


Synthesis and catalytic application of magnesium complexes bearing pendant indolyl ligands.
Three novel indole-based ligand precursors [HIndPh(R), R = methoxy, HIndPh(OMe) (); thiomethoxy, HIndPh(SMe) (); and N,N'-dimethylamino, HIndPh(NMe2) ()] have been synthesized via Sonogashira and cyclization reactions with moderate to high yield. Reactions of these ligand precursors with 0.6 equivalent of Mg(n)Bu2 in THF afforded the magnesium bis-indolyl complexes , respectively. All the ligand precursors and related magnesium complexes have been characterized by NMR spectroscopy and elemental analyses. The molecular structures are reported for compounds and . Under optimized conditions, compound demonstrates efficient catalytic activities towards the ring opening polymerization of l-lactide and ε-caprolactone in the presence of BnOH.


Dynamic differentiation of activated human peripheral blood CD8+ and CD4+ effector memory T cells.
Two functionally different memory T cell subsets were originally defined based on their different CCR7 expression profile, but the lineage relationship between these subsets referred to as central memory T cells (T(CM)) and effector memory T cells (T(EM)), is not resolved. A prevalent model proposes a linear progressive differentiation from T(CM) to T(EM). Our results demonstrate that on activation, human CCR7-CD62L- peripheral blood CD8+ and CD4+ T(EM) cells exhibit a dynamic differentiation, involving transient as well as stable changes to T(CM) phenotype and properties. Whereas the larger fraction of T(EM) cells increases expression of effector molecules, such as perforin or IFN-gamma, a smaller fraction first acquires CCR7 expression. We demonstrate that this acquisition of lymph node homing potential is associated with strong proliferation similar to that of activated T(CM) cells. After proliferation, most of these cells lose CCR7 expression again and acquire effector functions (e.g., perforin production). A small proportion (approximately 6%), however, maintain phenotypic and functional T(CM) properties over a long time interval. These results suggest that T(EM) cells provide immediate effector function by a fraction of cells as well as self-renewal by others through up-regulation of CCR7 followed by either secondary peripheral effector function or long term maintenance of T(CM)-like properties.


Systematic analysis of coproporphyrinogen oxidase gene defects in hereditary coproporphyria and mutation update.
Hereditary coproporphyria (HC) is an acute hepatic porphyria with autosomal dominant inheritance caused by deficient activity of coproporphyrinogen III oxidase (CPO). Clinical manifestations of the disease are characterized by acute attacks of neurological dysfunction often precipitated by drugs, fasting, cyclical hormonal changes, or infectious diseases. Skin photosensitivity may also be present. The seven exons, the exon/intron boundaries and part of 3' noncoding sequence of the CPO gene were systematically analyzed by an exon-by-exon denaturing gradient gel electrophoresis (DGGE) strategy followed by direct sequencing in seven unrelated heterozygous HC patients from France, Holland, and Czech Republic. Seven novel mutations and two new polymorphisms were detected. Among these mutations: two are missense (G197W, W427R), two are nonsense (Q306X, Q385X), two are small deletions (662de14bp; 1168del3bp removing a glycine at position 390), and one is a splicing mutation (IVS1-15c-->g) which creates a new acceptor splice site. The pathological significance of the point mutations G197W, W427R, and the in-frame deletion 390delGly were assessed by their respective expression in a prokaryotic system using site-directed mutagenesis. These mutations resulted in the absence or a dramatic decrease of CPO activity. The two polymorphisms were localized in noncoding part of the gene: 1) a C/G polymorphism in the promotor region, 142 bp upstream from the transcriptional initiation site (-142C/G), and 2) a 6 bp deletion polymorphism in the 3' noncoding part of the CPO gene, 574 bp downstream of the last base of the normal termination codon (+574 delATTCTT). Five intragenic dimorphisms are now well characterized and the high degree of allelic heterogeneity in HC is demonstrated with seven new different mutations making a total of nineteen CPO gene defects reported so far.


Comparison of the API 20E and BBL Crystal E/NF identification systems for differentiating bacterial isolates from apparently healthy reared sea bass (Dicentrarchus labrax).
The ability of two commercial rapid identification systems, API 20E and BBL Crystal E/NF, to reliably identify bacterial isolates from the internal organs of reared sea bass were compared. The tests gave different results: API 20E identified bacteria as Pseudomonas spp. with 37% accuracy, while BBL Crystal E/NF identified them as Flavobacterium odoratum with 99% accuracy. Although F. odoratum is not a marine fish pathogen, conventional tests conducted with the same isolates were more indicative of them being Flavobacterium spp. than Pseudomonas spp., suggesting that BBL Crystal E/NF was more reliable in this identification. Both systems were found to be applicable for diagnostics of marine fish pathogens, but should be used with caution because of possible misinterpretation.


Pediatric Cancer.
In pediatric brain tumors, the intensification of chemotherapy has allowed for a reduction in radiotherapy (RT) volume to an involved field approach, particularly in patients with medulloblastoma. For patients with low-grade gliomas, the trend has remained to delay RT with chemotherapy; however, when RT is used, typically smaller clinical target volume margins are used. For patients with extracranial tumors, intensive chemotherapy to address systemic disease with local control is considered standard. Proton beam therapy shows significant promise in addressing both short-term and long-term toxicities in both central nervous system (CNS) and non-CNS pediatric tumors.


Characterization of reciprocal translocations in pigs using dual-colour chromosome painting and primed in situ DNA labelling.
We report the use of dual-colour chromosome painting to determine the exact nature of certain chromosome rearrangements observed in the pig (Sus scrofa domestica). The chromosomal abnormalities were detected by GTG- and RBG-banding techniques. The initially proposed interpretations were: (1) rcp(6;13)(p1.5;q4.1); (2) rcp(11;16)(p1.4;q1.4); (3) rcp(6;16)(p1.1;q1.1); (4) rcp(13;17)(q4.1;q1.1); (5) rcp(6;14)(q2.7;q2.1); (6) rcp(3;5)(p1.3;q2.3); (7) rcp(2; 14)(q1.3;q2.7); (8) rcp(15;17)(q1.3;q2.1). Hybridizations were carried out with biotin- and digoxigenin-labelled probes obtained by priming authorizing random mismatches polymerase chain reaction (PARM-PCR) amplification of porcine flow-sorted chromosomes. In some cases, i.e. (1), (4), (5), (6), (7) and (8), the fluorescence in situ hybridization (FISH) results allowed confirmation of the interpretations proposed with classical cytogenetic methods. Chromosome painting proved the reciprocity of the translocation in cases (1), (6) and (8), whereas modifications of the formula were proposed for case (2). Primed in situ DNA labelling (PRINS) experiments have also been carried out in case (3) using a primer specific for the centromeres of acrocentric chromosomes (first experiment) or a primer specific for the centromeres of a subset of meta- and submetacentric chromosomes including chromosome 6 (second experiment). It allowed us to demonstrate that the breakpoints occurred in the centromeric region of chromosome 16 and in the p. arm of chromosome 6, just above the centromere.


Effects of a motivational, individual and locally anchored exercise intervention (MILE) on cardiorespiratory fitness: a community-based randomised controlled trial.
Risk factors for chronic disease, including low cardiorespiratory fitness levels (VO2max), are often present in middle-aged populations. We aimed to evaluate the efficacy of a motivational, individual, and locally anchored physical activity intervention on increasing VO2max in 30-49 year-olds with low VO2max. 232 adult volunteers with low VO2max were randomised to intervention (n = 115) or routine care (n = 117). The intervention included four motivational interviews; six months' free membership to a local sports club; and a GPS-watch/activity monitor for uploading training data to an online platform/community. Routine care was one or two motivational interviews. Inclusion criteria were low VO2max based on the cut off levels: ≤ 39 and ≤ 35 ml O2/kg/min. For 30-39 and 40-49 year-old men respectively and ≤ 33 and ≤ 31 ml O2/kg/min for 30-39 and 40-49 year-old women, respectively. The primary outcome was change in VO2max from baseline to twelve months estimated with a maximal ergometer bicycle test. Secondary outcomes included physical activity, biochemical and anthropometric measures, and health-related quality of life. The primary analyses were based on all available data and sensitivity- and predefined sex analyses were performed. The between-group differences were estimated using independent t-tests and presented with 95% confidence intervals. No significant between-group differences in primary or secondary outcomes were found at twelve months' follow-up. The mean VO2max change from baseline in the intervention- and routine care (ml/kg/min) was 3.8 (95% CI: 2.6; 5.0) and 3.4 (95% CI: 1.7; 5.2), respectively. No changes in physical activity were observed. The mean VO2max (ml/kg/min) changes from baseline in the intervention- and routine care group in men were: 5.0 (95% CI: 3.5; 6.4) and 3.5 (95% CI: 1.5; 5.4); and in women: 1.5 (95% CI: -0.1; 3.1) and 3.4 (95% CI: -0.1; 7.8), respectively. Significant differences in VO2max between non-completers (44.2%) and completers were observed, 26.3 (95% CI: 25.1; 27.5) vs 28.2 ml/kg/min (95% CI: 27.1; 29.0; p = 0.02). Sensitivity analyses did not change the main result. Offering a multi-component physical activity intervention to 30-49 year-olds with low levels of VO2max had no effect on the change in VO2max from baseline to twelve months compared with routine care. ClinicalTrials.gov (no. NCT01801956 ). Registered 1 March 2013.


Optimal K-Wire Placement for Indirect Cannulated Screw Fixation of Coronoid Process Fractures: A Radiology-Based Guide.
Indirect fixation of coronoid process fractures of the ulna, especially arthroscopically, offers the possibility to avoid extensive medial surgical approaches. However, optimal placement of K-wires for cannulated screw fixation is technically challenging. The aim of the present study was to present guide values for K-wire placement for indirect screw fixation of coronoid tip (COT) and anteromedial facet (AMF) fractures of the ulna. Computed tomography scans of the elbows of 197 patients with an uninjured ulna were identified and evaluated following a standard measuring protocol. Optimal placement of K-wires was defined as the bisector between the anterior and articular cortex while respecting a 1.6-mm safe zone for each placement. This placement of the K-wires in the COT and the AMF was analyzed by measuring the wire angulation in relation to the posterior ulnar cortex (AUC), the distance from the entry point to the posterior olecranon edge (DPE), and the intraosseous length (IOL). Because the coronoid has a curved shape, measurements of the COT and AMF were expected to differ significantly. Sex was also expected to influence measurements. To determine whether this optimal placement of K-wires can be reliably identified, interobserver and intraobserver reliabilities were evaluated. To address tip fragments, we determined a mean AUC of 64°, DPE of 36 mm, and IOL of 38 mm. Regarding wire placement toward the AMF, the means for the AUC, DPE, and IOL were 79°, 27 mm, and 33 mm, respectively. Statistically significant differences were found between the COT and the AMF for the AUC, DPE, and IOL. Our measuring protocol showed good interobserver and intraobserver reliability. K-wire placement toward the COT demands a smaller angle, a longer IOL, and a more distal entry point than wires placed toward AMF. These differences reflect the curved and complex anatomy of the coronoid. This measuring algorithm can be used to plan osteosynthesis, and the obtained reference values can help understand the coronoid's complex anatomy.


ATR-FTIR spectroscopy in blood plasma combined with multivariate analysis to detect HIV infection in pregnant women.
The primary concern for HIV-infected pregnant women is the vertical transmission that can occur during pregnancy, in the intrauterine period, during labour or even breastfeeding. The risk of vertical transmission can be reduced by early diagnosis. Therefore, it is necessary to develop new methods to detect this virus in a quick and low-cost fashion, as colorimetric assays for HIV detection tend to be laborious and costly. Herein, attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy combined with multivariate analysis was employed to distinguish HIV-infected patients from healthy uninfected controls in a total of 120 blood plasma samples. The best sensitivity (83%) and specificity (92%) values were obtained using the genetic algorithm with linear discriminant analysis (GA-LDA). These good classification results in addition to the potential for high analytical frequency, the low cost and reagent-free nature of this method demonstrate its potential as an alternative tool for HIV screening during pregnancy.


Radiotherapy for oral cavity cancers.
Intensity modulated radiation therapy and brachytherapy are standard techniques of irradiation for the treatment of oral cavity cancers. These techniques are detailed in terms of indication, planning, delineation and selection of the volumes of interest, dosimetry and patients positioning control. This is an update of the guidelines of the French Society of Radiotherapy Correspondence.