Failure of observing NeuroD1-induced microglia-to-neuron conversion in vitro is not attributed to the low NeuroD1 expression level.
NeuroD1-induced microglia-to-neuron conversion is hotly debated. Recently, we published a paper in Neuron demonstrating that NeuroD1 cannot induce microglia-to-neuron cross-lineage conversion. In the same issue of Neuron, Matsuda et al., who observed the "NeuroD1-induced microglia-to-neuron conversion" phenotype, responded to our study. They claimed that we failed to observe NeuroD1-induced microglia-to-neuron conversion in vitro due to the low NeuroD1 expression efficiency in our experiment. They argued that the NeuroD1 upregulation in our study was around 200-fold (vs. control), whereas the upregulation in Nakashima lab was 3000-fold, 15 times higher than ours. In fact, this is not true. We compared the expression level from the original paper and found that our NeuroD1 expression level was comparable to that of Matsuda et al. (Neuron 101:472-485.e477, 2019), or even higher. Therefore, the failure of observing NeuroD1-induced microglia-to-neuron conversion cannot be attributable to the low expression level.


Reflex oral activity in decerebrate rats of different age.
Rats decerebrated at different ages (4 days to greater than 100 days) were tested for a rhythmic reflex 'suckling' response and for a simple digastric reflex. The ease with which the 'suckling' response could be elicited decreased with increasing age; the time course of this change approximated to that of changing characteristics of the digastric reflex. The mean latency of the simple reflex response decreased non-linearly from 70 msec in the 4 day postpartum animal to 5 msec in the adult animal; the duration of the response also decreased from 25 msec to 10 msec but this change took place mainly between 8 and 15 days postpartum. The latency of the digastric response also shortened in animals decerebrated on day 4 and kept for 2 days. It was concluded that the observed changes in the digastric reflex occurred independently of encephalic structures.


The PseEF efflux system is a virulence factor of Pseudomonas syringae pv. syringae.
An ATP-binding cassette (ABC) transporter, called the PseEF efflux system, was identified at the left border of the syr-syp genomic island of Pseudomonas syringae pv. syringae strain B301D. The PseEF efflux system was located within a 3.3-kb operon that encodes a periplasmic membrane fusion protein (PseE), and an ABC-type cytoplasmic membrane protein (PseF). The PseEF efflux system exhibited amino acid homology to a putative ABC efflux system (MacAB) of E. coli W3104 with identities of 47.2% (i.e., PseE to MacA) and 57.6% (i.e., PseF to MacB). A nonpolar mutation within the pseF gene was generated by nptII insertional mutagenesis. The resultant mutant strain showed significant reduction in secretion of syringomycin (74%) and syringopeptin (71%), as compared to parental strain B301D. Quantitative real-time RT-PCR was used to determine transcript levels of the syringomycin (syrB1) and syringopeptin (sypA) synthetase genes in strain B301D-HK7 (a pseF mutant). Expression of the sypA gene by mutant strain B301D-HK7 was approximately 6.9% as compared to that of parental strain B301D, while the syrB1 gene expression by mutant strain B301D-HK7 was nearly 14.6%. In addition, mutant strain B301D-HK7 was less virulent by approximately 67% than parental strain B301D in immature cherry fruits. Mutant strain B301D-HK7 was not reduced in resistance to any antibiotics used in this study as compared to parental strain B301D. Expression (transcript levels) of the pseF gene was induced approximately six times by strain B301D grown on syringomycin minimum medium (SRM) supplemented with the plant signal molecules arbutin and D-fructose (SRMAF), as compared to that of strain B301D grown on SRM (in the absence of plant signal molecules). In addition, during infection of bean plants by P. syringae pv. syringae strain B728a, expression of the pseF gene increased at 3 days after inoculation (dai). More than 180-fold induction was observed in transcript levels of the pseF gene by parental strain B728a as compared to strain B728a-SL7 (a salA mutant). Thus, the PseEF efflux system, an ABC-type efflux system, has an important role in secretion of syringomycin and syringopeptin, and is required for full virulence in P. syringae pv. syringae.


Local complications of ascending phlebography.
In order to estimate the rate of local complications of phlebography we studied a one-year series of 477 patients. Half of the patients were examined with a comparatively highly concentrated contrast medium and half with a more diluted one. Eight patients had local complications, verified by biopsy and/or thermography and plethysmography. One had a deep vein thrombosis, five had a superficial thrombosis and two had a skin necrosis. There were more complications after the use of the more concentrated medium, six against two. The frequency of clinically significant complications to phlebography is rather low, but high enough to warrant attention to the risks of the method. The concentration of the contrast medium should be as low as possible, and the time that the vessels are exposed to the chemical trauma as short as possible. A hypothesis is brought forward that skin necrosis with slow healing could be secondary to arterial thrombosis caused by extravasal contrast injection.


Mitochondria-wide association study observed significant interactions of mitochondrial respiratory and the inflammatory in the development of anxiety and depression.
The aim of this study was to investigate the possible interaction of mitochondrial dysfunction and inflammatory cytokines in the risk of anxiety and depression. We utilized the UK Biobank for the sample of this study. A mitochondria-wide association(MiWAS) and interaction analysis was performed to investigate the interaction effects of mitochondrial DNA (mtDNA)×C-reactive protein (CRP) on the risks of self-reported anxiety (N = 72,476), general anxiety disorder (GAD-7) scores (N = 80,853), self-reported depression (N = 80,778), Patient Health Questionnaire (PHQ-9) scores (N = 80,520) in total samples, females and males, respectively, adjusting for sex, age, Townsend deprivation index (TDI), education score, alcohol intake, smoking and 10 principal components. In all, 25 mtSNPs and 10 mtSNPs showed significant level of association with self-reported anxiety and GAD-7 score respectively. A total of seven significant mtDNA × CRP interactions were found for anxiety, such as m.3915G>A(MT-ND1) for self-reported anxiety in total subjects (P = 6.59 × 10-3), m.4561T>C(MT-ND2) (P = 3.04 × 10-3) for GAD-7 score in total subjects. For depression, MiWAS identified 17 significant mtSNPs for self-reported depression and 14 significant mtSNPs for PHQ-9 scores. 17 significant mtDNA associations (2 for self-reported depression and 15 for PHQ-9 score) was identified, such as m.14869G>A(MT-CYB; P = 2.22 × 10-3) associated with self-reported depression and m.4561T>C (MT-ND2; P value = 3.02 × 10-8) associated with PHQ-9 score in all subjects. In addition, 5 common mtDNA shared with anxiety and depression were found in MiWAS, and 4 common mtDNA variants were detected to interact with CRP for anxiety and depression, such as m.9899T>C(MT-CO3). Our study suggests the important interaction effects of mitochondrial function and CRP on the risks of anxiety and depression.


Chronic total occlusion is associated with a higher incidence of malapposition and uncovered stent struts: OCT findings at 6 months following DES implantation.
To compare stent coverage and malapposition in patients with chronic total occlusion (CTO) lesions and non-CTO lesions (including lipid-rich plaque [LRP] and non-lipid-rich plaque [non-LRP]) after drug-eluting stent (DES) implantation by optical coherence tomography (OCT). Different initial lesion characteristics may be related to heterogeneous vessel responses after DES implantation. However, the vessel response in patients with CTO and non-CTO lesions after stenting is unclear. Methods We retrospectively enrolled 64 patients with 68 target lesions. All of the patients underwent OCT imaging immediate after stenting and 6 months after stenting. LRP was defined as the plaque with lipid content in ≥2 quadrants. Non-LRP consisted of fibrous, fibrocalcific plaque, and lipid plaque with less than 2 quadrants lipid content. The malapposition (3.0%, 2.6% vs. 0.6%, P = 0.022), tissue protrusion (15.0% vs. 11.0% vs. 6.4%, P < 0.001), and intrastent thrombus (3.8% vs. 2.4% vs. 1.1%, P = 0.012) were more frequent in the CTO and LRP groups. At 6-month follow-up, malapposition (5.0% vs. 1.0% and 0.4%, P = 0.002) and cross sections with uncovered struts (23.4% vs. 8.2% and 6.6%, P < 0.001) were most frequently observed in the CTO group. Although the incidence of stent thrombosis was non-significantly higher in the CTO group than the other two groups, no events were observed in patients with CTO. Patients with CTO lesions showed unfavorable responses to DES in the acute phase as well as at the 6-month follow-up, indicating the important pathological link between the original lesion morphology underneath the stents and heterogeneous artery healing. © 2017 Wiley Periodicals, Inc.


Extraction, structural-activity relationships, bioactivities, and application prospects of Bletilla striata polysaccharides as ingredients for functional products: A review.
Bletilla striata is a well-known medicinal plant with high pharmaceutical and ornamental values. Polysaccharide is the most important bioactive ingredient in B. striata and has various health benefits. Recently, B. striata polysaccharides (BSPs) have attracted much attention from industries and researchers due to their remarkable immunomodulatory, antioxidant, anti-cancer, hemostatic, anti-inflammatory, anti-microbial, gastroprotective, and liver protective effects. Despite the successful isolation and characterization of BSPs, there is still limited knowledge regarding their structure-activity relationships (SARs), safety concerns, and applications, which hinders their full utilization and development. Herein, we provided an overview of the extraction, purification, and structural features, as well as the effects of different influencing factors on the components and structures of BSPs. We also highlighted and summarized the diversity of chemistry and structure, specificity of biological activity, and SARs of BSP. The challenges and opportunities of BSPs in the food, pharmaceutical, and cosmeceutical fields are discussed, and the potential development and future study direction are scrutinized. This article provides comprehensive knowledge and underpinnings for further research and application of BSPs as therapeutic agents and multifunctional biomaterials.


Comparison of lysine and tryptophan catabolizing enzymes in rat and bovine tissues.
Earlier studies indicate that alpha-aminoadipate aminotransferase (AadAT) and kynurenine aminotransferase (KAT) activities from rat tissues are associated with a single protein. However, our recent studies indicate that AadAT activity from bovine liver and kidney is not associated with KAT activity. To test whether the lysine and tryptophan catabolism in bovine tissues differ from that in rat tissues, we compared the activities of enzymes involved in lysine and tryptophan pathways in rat and bovine tissues. The activities of lysine catabolizing enzymes such as AadAT, lysine alpha-ketoglutarate reductase and saccharopine dehydrogenase in the bovine tissues were significantly lower than those found in rat tissues. The activities of tryptophan catabolizing enzymes such as KAT and kynurenine hydroxylase in the bovine tissues were negligible as compared to those in rat tissues. The results suggest that lysine is degraded via the saccharopine pathway in the livers and kidneys of both species but the metabolism of tryptophan in bovine tissues may be different from that in rat tissues.


Parenteral nutrition in short bowel syndrome patients, regardless of its duration, increases serum proinflammatory cytokines.
Short bowel syndrome is a severe malabsorption disorder, and prolonged parenteral nutrition is essential for survival in some cases. Among the undesirable effects of long-term parenteral nutrition is an increase in proinflammatory cytokines. The aim of the present study was to measure the serum levels of interleukin-6, interleukin-10, tumor necrosis factor alpha, and transforming growth factor beta, in patients with short bowel syndrome on cyclic parenteral nutrition and patients who had previously received but no longer require parenteral nutrition. The study was cross-sectional and observational. Three groups were studied as follows: Parenteral nutrition group, 9 patients with short bowel syndrome that receive cyclic parenteral nutrition; Oral nutrition group, 10 patients with the same syndrome who had been weaned off parenteral nutrition for at least 1 year prior to the study; Control group, 13 healthy adults, matched for age and sex to parenteral and oral groups. The following data were collected: age, tobacco use, drug therapies, dietary intake, body weight, height, blood collection. All interleukins were significantly higher in the parenteral group compared with the control group as follows: interleukin-6: 22 ± 19 vs 1.5 ± 1.4 pg/mL, P= .0002; transforming growth factor β: 854 ± 204 vs 607 ± 280 pg/mL, P= .04; interleukin-10: 8 ± 37 vs 0.6 ± 4, P= .03; tumor necrosis factor α: 20 ± 8 vs 8 ± 4 pg/mL, P< .0001. We concluded that parenteral nutrition in short bowel syndrome patients, regardless of its duration, increases serum proinflammatory cytokines.


cGAS-STING pathway aggravates early cerebral ischemia-reperfusion injury in mice by activating NCOA4-mediated ferritinophagy.
Stroke patients are often complicated by cerebral ischemia-reperfusion injury (CIRI) after the restoration of cerebral perfusion, and how to prevent CIRI at an early stage has received close attention. The imbalance of iron metabolism is one of the essential factors in the aggravation of CIRI, and NCOA4-mediated ferritinophagy, as a critical pathway to regulate iron metabolism, is expected to be an effective intervention target. We established a mouse model of cerebral ischemia-reperfusion (CIR) with NCOA4 silencing. We found that activation of NCOA4-mediated ferritinophagy atthe early stage of CIR mediated the onset of oxidative stress and contributed to autophagy and apoptosis, and eventually resulted in increased brain injury. This suggests that NCOA4-mediated ferritinophagy plays a vital role in early CIR and can be an effective target to prevent and treat CIRI. We next explored the upstream regulatory targets of NCOA4-mediated ferritinophagy. The previous evidence for the cGAS-STING pathway's importance during CIR and its strong relationship with autophagy attracted our attention. To investigate whether the cGAS-STING pathway regulates NCOA4-mediated ferritinophagy, we further administered a cGAS inhibitor to mice with CIR and overexpressed NCOA4. Along with the inhibition of the cGAS-STING pathway, ferritinophagy, oxidative stress, autophagy, and apoptosis were inhibited, and CIRI was ameliorated, which was attenuated by NCOA4 overexpression. In conclusion, our results suggest that activation of the cGAS-STING pathway exacerbates CIRI at the early stage of CIR, which may be achieved by mediating NCOA4-mediated ferritinophagy.


Functional characterization of Kaposi's sarcoma-associated herpesvirus small capsid protein by bacterial artificial chromosome-based mutagenesis.
A systematic investigation of interactions amongst KSHV capsid proteins was undertaken in this study to comprehend lesser known KSHV capsid assembly mechanisms. Interestingly the interaction patterns of the KSHV small capsid protein, ORF65 suggested its plausible role in viral capsid assembly pathways. Towards further understanding this, ORF65-null recombinant mutants (BAC-∆65 and BAC-stop65) employing a bacterial artificial chromosome (BAC) system were generated. No significant difference was found in both overall viral gene expression and lytic DNA replication between stable monolayers of 293T-BAC36 (wild-type) and 293T-BAC-ORF65-null upon induction with 12-O-tetradecanoylphorbol-13-acetate, though the latter released 30-fold fewer virions to the medium than 293T-BAC36 cells. Sedimentation profiles of capsid proteins of ORF65-null recombinant mutants were non-reflective of their organization into the KSHV capsids and were also undetectable in cytoplasmic extracts compared to noticeable levels in nuclear extracts. These observations collectively suggested the pivotal role of ORF65 in the KSHV capsid assembly processes.


Preparation and characterization of biocompatible chitosan nanoparticles for targeted brain delivery of peptides.
Here, we describe a nanocarrier system that can transfer chitosan nanoparticles loaded with either small peptides such as the caspase inhibitor Z-DEVD-FMK or a large peptide like basic fibroblast growth factor across the blood-brain barrier. The nanoparticles are selectively directed to the brain and are not measurably taken up by liver and spleen. Intravital fluorescent microscopy provides an opportunity to study the penetration kinetics of nanoparticles loaded with fluorescent agents such as Nile red, and has demonstrated that this nanomedicine formulation is rapidly transported across the blood-brain barrier.


Longitudinal development of specific and functional antibody in very low birth weight premature infants.
We evaluated the formation of specific and functional antibody in preterm infants born weighing less than 1500 g (mean 1088 g) and less than 32 wk gestational age (mean 28.8 wk). Plasma IgG antibody against tetanus and diphtheria toxoids were measured by an enzyme-linked immunosorbent assay. Opsonic activity of heat-inactivated plasma was measured using radiolabeled bacteria, adult polymorphonuclear leukocytes and exogenous human complement. In the presence of complement, the strain of coagulase negative staphylococcus used was opsonized by IgG antibody, and the strain of Escherichia coli by IgM. Geometric mean plasma levels of tetanus and diphtheria IgG antibody fell from birth to 4 months chronological age, but rose significantly by 9 months (approximately 2 months after the third dose of diphtheria, tetanus, pertussis vaccine). However, at 9 months they remained lower than the respective geometric mean levels in 9-month-old term infants (tetanus: p less than 0.001; diphtheria: p = 0.02). The preterm infants' mean plasma IgG staphylococcal opsonic activity fell from birth to 2.5 months, but by 9 months was comparable to that of term infants of the same age. Mean IgM opsonic activity for E. coli was very low at birth in both preterm and term infants. It rose with chronological age, correlating with the rise in total IgM (r = 0.48, p less than 0.001) and by 9 months the mean preterm and term infants' levels of IgM opsonic activity for E. coli were comparable.(ABSTRACT TRUNCATED AT 250 WORDS)


Vasoactive intestinal polypeptide and carbachol act synergistically to induce the hydrolysis of inositol containing phospholipids in the rat superior cervical ganglion.
The effects of vasoactive intestinal polypeptide (VIP) and carbachol on inositol lipid breakdown were assayed in isolated rat superior cervical ganglia. We report here that VIP and carbachol act synergistically to stimulate the formation of inositol phosphates. This synergistic interaction may explain the modulatory effect of VIP on the muscarinic transmission in the sympathetic ganglia.


Extinction Learning Across Development: Neurodevelopmental Changes and Implications for Pediatric Anxiety Disorders.
Alterations in extinction learning relate to the development and maintenance of anxiety disorders across the lifespan. While exposure therapy, based on principles of extinction, can be highly effective for treating anxiety, many patients do not show sufficient improvement following treatment. In particular, evidence suggests that exposure therapy does not work sufficiently for up to 40% of children who receive this evidence-based treatment.Importantly, fear learning and extinction, as well as the neural circuitry supporting these processes, undergo dynamic changes across development. An improved understanding of developmental changes in extinction learning and the associated neural circuitry may help to identify targets to improve treatment response in clinically anxious children and adolescents. In this chapter, we provide a brief overview of methods used to study fear learning and extinction in developmental populations. We then review what is currently known about the developmental changes that occur in extinction learning and related neural circuitry. We end this chapter with a discussion of the implications of these neurodevelopmental changes for the characterization and treatment of pediatric anxiety disorders.


Electron Oscillation-Induced Splitting Electroluminescence from Nano-LEDs for Device-Level Encryption.
Data security is a major concern in digital age, which generally relies on algorithm-based mathematical encryption. Recently, encryption techniques based on physical principles are emerging and being developed, leading to the new generation of encryption moving from mathematics to the intersection of mathematics and physics. Here, device-level encryption with ideal security is ingeniously achieved using modulation of the electron-hole radiative recombination in a GaN-light-emitting diode (LED). When a nano-LED is driven in the non-carrier injection mode, the oscillation of confined electrons can split what should be a single light pulse into multiple pulses. The morphology (amplitude, shape, and pulse number) of those history-dependent multiple pulses that act as carriers for transmitted digital information depends highly on the parameters of the driving signals, which makes those signals mathematically uncrackable and can increase the volume and security of transmitted information. Moreover, a hardware and software platform are designed to demonstrate the encrypted data transmission based on the device-level encryption method, enabling recognition of the entire ASCII code table. The device-level encryption based on splitting electroluminescence provides an encryption method during the conversion process of digital signals to optical signals and can improve the security of LED-based communication.


Subchronic milnacipran treatment increases basal extracellular noradrenaline concentrations in the medial prefrontal cortex of rats.
In this study, we investigated the acute effects of milnacipran, a serotonin-noradrenaline reuptake inhibitor, following subchronic treatment with milnacipran (30 mg/kg periorally for 7 days) on extracellular noradrenaline, dopamine and serotonin concentrations in the medial prefrontal cortex. Subchronic administration of milnacipran produced significantly higher basal levels of extracellular noradrenaline. Acute milnacipran administration following subchronic milnacipran treatment for 7 days produced a greater increase in extracellular noradrenaline than a single dose of milnacipran alone. The present results suggest that subchronic milnacipran treatment enhances noradrenergic neural transmission beyond that achieved with acute administration of milnacipran alone, but has no effect on serotonergic or dopaminergic neural transmission.


Robotic McKeown esophagectomy with recurrent laryngeal nerve lymph node dissection: how I do it.
We describe our standardized approach to robotic esophagectomy with special emphasis on the technical aspects pertaining to bilateral recurrent laryngeal nerve lymph node dissection.


A case of Currarino syndrome in an adult female presenting with refractory chronic constipation.
We present the case of a 25-year-old female who presented due to refractory chronic constipation and fecal incontinence. She had bowel movements every 7-30 days with an increased consistency (1-2 Bristol type stools), together with soiling and passive fecal incontinence (Wexner Scale: 12/20). She had previously undergone surgery shortly after birth for an anorectal malformation repair. The colonoscopy and histological study of the rectum were normal. A pelvic magnetic resonance imaging (MRI) was performed, which showed a right pararectal mass that compressed the rectum without invading it. This mass was compatible with a presacral teratoma or hamartoma. MRI also revealed coccyx agenesis and hypoplasia of the last sacral vertebrae (Image 1), which were consistent with Currarino syndrome (CS). The patient received 14 sessions of transcutaneous electrostimulation of the posterior tibial nerve, resulting in an increase in bowel movements (every 3 days) and a reduction in fecal incontinence. She was then referred to surgery for presacral mass removal.


[New Pathogenetic Concepts and Early Pharmacological Studies in Sarcoidosis].
The etiology of sarcoidosis is still elusive, yet there has been considerable progress in various areas of basic and clinical research. This review focuses on mechanisms of granuloma formation and on new findings in autoimmunity and genetics of sarcoidosis. A new promising concept arose, where serum amyloid A and/or mycobacterial antigens serve as nidus for granuloma formation. Furthermore, autoimmunity in sarcoidosis was neglected for a long time, yet new studies found autoantigens and abnormalities in antigen presentation in sarcoidosis. Last but not least, large genome-wide association studies discovered several new predisposing genes, leading to new hypotheses on pathomechanisms of sarcoidosis.In the second part, we focus on ongoing or recently completed clinical-pharmacological studies in patients with sarcoidosis: Positive studies were published in well characterized and homogenous subcohorts of sarcoid patients. Several drugs have shown a positive effect on sarcoidosis-associated fatigue, on sarcoidosis of the skin and on pulmonary hypertension in sarcoid patients. It seems that the generation of clinically closely defined subcohorts is necessary to achieve positive outcomes in studies on sarcoidosis.


Discovery of 4-aminoindole carboxamide derivatives to curtail alpha-synuclein and tau isoform 2N4R oligomer formation.
Alzheimer's disease (AD) is a multifactorial, chronic neurodegenerative disease characterized by the presence of extracellular β-amyloid (Aβ) plaques, intraneuronal neurofibrillary tangles (NFTs), activated microglial cells, and an inflammatory state (involving reactive oxygen species production) in the brain. NFTs are comprised of misfolded and hyperphosphorylated forms of the microtubule-binding protein tau. Interestingly, the trimeric form of the 2N4R splice isoform of tau has been found to be more toxic than the trimeric 1N4R isoform in neuron precursor cells. Few drug discovery programs have focused on specific tau isoforms. The present drug discovery project is centered on the anti-aggregation effect of a series of seventeen 4- or 5-aminoindole carboxamides on the 2N4R isoform of tau. The selection of the best compounds was performed using α-synuclein (α-syn). The anti-oligomer and -fibril activities of newly synthesized aminoindole carboxamide derivatives were evaluated with biophysical methods, such as thioflavin T fluorescence assays, photo-induced cross-linking of unmodified proteins, and transmission electron microscopy. To evaluate the reduction of inclusions and cytoprotective effects, M17D neuroblastoma cells expressing inclusion-forming α-syn were treated with the best amide representatives. The 4-aminoindole carboxamide derivatives exhibited a better anti-fibrillar activity compared to their 5-aminoindole counterparts. The amide derivatives 2, 8, and 17 exerted anti-oligomer and anti-fibril activities on α-syn and the 2N4R isoform of tau. At a concentration of 40 μM, compound 8 reduced inclusion formation in M17D neuroblastoma cells expressing inclusion-prone αSynuclein3K::YFP. Our results demonstrate the potential of 4-aminoindole carboxamide derivatives with regard to inhibiting the oligomer formation of α-syn and tau (2N4R isoform) for further optimization prior to pre-clinical studies.


Evaluation of Digital Health Strategy to Support Clinician-Led Critically Ill Patient Population Management: A Randomized Crossover Study.
To investigate whether a novel acute care multipatient viewer (AMP), created with an understanding of clinician information and process requirements, could reduce time to clinical decision-making among clinicians caring for populations of acutely ill patients compared with a widely used commercial electronic medical record (EMR). Single center randomized crossover study. Quaternary care academic hospital. Attending and in-training critical care physicians, and advanced practice providers. AMP. We compared ICU clinician performance in structured clinical task completion using two electronic environments-the standard commercial EMR (Epic) versus the novel AMP in addition to Epic. Twenty subjects (10 pairs of clinicians) participated in the study. During the study session, each participant completed the tasks on two ICUs (7-10 beds each) and eight individual patients. The adjusted time for assessment of the entire ICU and the adjusted total time to task completion were significantly lower using AMP versus standard commercial EMR (-6.11; 95% CI, -7.91 to -4.30 min and -5.38; 95% CI, -7.56 to -3.20 min, respectively; p < 0.001). The adjusted time for assessment of individual patients was similar using both the EMR and AMP (0.73; 95% CI, -0.09 to 1.54 min; p = 0.078). AMP was associated with a significantly lower adjusted task load (National Aeronautics and Space Administration-Task Load Index) among clinicians performing the task versus the standard EMR (22.6; 95% CI, -32.7 to -12.4 points; p < 0.001). There was no statistically significant difference in adjusted total errors when comparing the two environments (0.68; 95% CI, 0.36-1.30; p = 0.078). When compared with the standard EMR, AMP significantly reduced time to assessment of an entire ICU, total time to clinical task completion, and clinician task load. Additional research is needed to assess the clinicians' performance while using AMP in the live ICU setting.


Distinct monitoring strategies underlie costs and performance in prospective memory.
Prospective memory (PM) describes the ability to remember to perform goal-relevant actions at an appropriate time in the future amid concurrent demands. A key contributor to PM performance is thought to be the effortful monitoring of the environment for PM-related cues, a process whose existence is typically inferred from a behavioral interference measure of reaction times. This measure, referred to as "PM costs," is an informative but indirect proxy for monitoring, and it may not be sufficient to understand PM behaviors in all situations. In this study, we asked participants to perform a visual search task with arrows that varied in difficulty while concurrently performing a delayed-recognition PM task with pictures of faces and scenes. To gain a precise measurement of monitoring behavior, we used eye-tracking to record fixations to all task-relevant stimuli and related these fixation measures to both PM costs and PM accuracy. We found that PM costs reflected dissociable monitoring strategies: higher costs were associated with early and frequent monitoring while lower costs were associated with delayed and infrequent monitoring. Moreover, the link between fixations and PM costs varied with cognitive load, and the inclusion of fixation data yielded better predictions of PM accuracy than using PM costs alone. This study demonstrates the benefit of eye-tracking to disentangle the nature of PM costs and more precisely describe strategies involved in prospective remembering.


The effect of propofol on chemosensitivity of paclitaxel in cervical cancer cells.
Propofol is a drug with potential anticancer effect. This study aimed to explore the effect of propofol on chemosensitivity of cervical cancer cells to paclitaxel. HeLa and CaSki cells were selected for drug experiments. Cell viability was evaluated via CCK-8 assay, and the combination index (CI) was calculated by CompuSyn software. A clinically relevant concentration and IC30 of propofol were selected in combination with 5 nM paclitaxel. BrdU incorporation, transwell, and flow cytometry assays were utilized to evaluate cell proliferation, migration, invasion, and apoptosis. The expression of β-tubulin, stathmin 1, and GAPDH proteins was evaluated by Western blot. The stathmin 1 cDNA plasmid was used to establish stathmin 1-overexpressing CaSki cells. At clinically relevant concentrations (0-80 μM), propofol did not affect cancer cell viability, but high concentrations (100-800 μM) reduced cell viability. The CI values of propofol with IC30 (200 μM in HeLa; 400 μM in CaSki) combined with 5 nM paclitaxel were <1. The effect of propofol with IC30 combined with paclitaxel on cell proliferation, migration, invasion, and apoptosis were stronger than individual effect, while 30 μM propofol had no effect. The Western blot results showed 30 μM propofol did not affect β-tubulin and stathmin 1 expression in cells, although paclitaxel upregulated β-tubulin expression while downregulating stathmin 1 expression. Compared with paclitaxel alone, cotreatment with propofol at its IC30 and paclitaxel decreased stathmin 1 expression but had no effect on β-tubulin expression. High stathmin 1 expression weakened the effect of paclitaxel on cell viability and apoptosis, while propofol partially reversed these effect. Propofol at clinically relevant concentrations had no effect on the malignant biological behaviors of cervical cancer cells, while propofol at high concentrations decreased.Propofol with IC30 and paclitaxel had synergetic effect on cancer cells through a reduction in stathmin 1 expression.


Michel dysplasia. Common cavity inner ear deformity.
A case is reported in whom there was a unilateral complete arrest of differentiation of the otocyst associated with an ipsilateral mild hemifacial hypoplasia and auricular abnormalities. The patient's tympanic membrane and VII nerve function were normal and a normal ossicular mass was identified on CT Scan. This case illustrates that development of the middle ear cavity (exclusive of structures on the medial wall), external ear canal and VII nerve can be independent of development of the inner ear and that a unilateral absence of otic capsule differentiation can take place without severe associated abnormalities.


Spatial distribution of illicit drugs in surface waters of the natural park of Pego-Oliva Marsh (Valencia, Spain).
The Pego-Oliva Marsh is the second most important wetland in the Valencian Community (Spain). It is included in the RAMSAR agreement and represents one key point for migratory birds. Emerging contaminants from the human pressure, such as pharmaceuticals, illicit drugs and personal care product, are not included in the list of priority contaminants of the Water Framework Directive yet, and are neither monitored nor controlled. However, pollution of emerging contaminants can threaten the environment and even human health. In order to understand the status of the emerging contamination and recommend future rationalization of countermeasures, the occurrence of illicit drugs was investigated. Samples were collected at 23 sites from the main irrigation channels and the marsh. Illicit drugs were extracted using solid phase extraction and determined by liquid chromatography tandem mass spectrometry. The method detection limits ranged from 0.01 to 1.54 ng l(-1) and the recoveries from 57% to 120%. 3,4-Methylenedioxymethamphetamine, ketamine, morphine, benzoylecgonine, cocaine, methadone, 6-acetylmorphine and nor-9-carboxy-tetrahydrocannabinol were detected. The mean concentrations were 0.62, 21.33, 1.30, 1.92, 2.25, 0.32, 0.04 and 0.07 ng l(-1), respectively. The highest concentrations were in the north of Pego-Oliva Marsh. The pollution status by illicit drugs of the Pego-Oliva Marsh has been established. However, contamination levels in all the area of the natural park were low compared with those reported in other superficial waters.


CD86⁺/CD206⁺, Diametrically Polarized Tumor-Associated Macrophages, Predict Hepatocellular Carcinoma Patient Prognosis.
Tumor-associated macrophages (TAMs), the most abundant infiltrating immune cells in tumor microenvironment, have distinct functions in hepatocellular carcinoma (HCC) progression. CD68⁺ TAMs represent multiple polarized immune cells mainly containing CD86⁺ antitumoral M1 macrophages and CD206⁺ protumoral M2 macrophages. TAMs expression and density were assessed by immunohistochemical staining of CD68, CD86, and CD206 in tissue microarrays from 253 HCC patients. Clinicopathologic features and prognostic value of these markers were evaluated. We found that CD68⁺ TAMs were not associated with clinicopathologic characteristics and prognosis in HCC. Low presence of CD86⁺ TAMs and high presence of CD206⁺ TAMs were markedly correlated with aggressive tumor phenotypes, such as multiple tumor number and advanced tumor-node-metastasis (TNM) stage; and were associated with poor overall survival (OS) (p = 0.027 and p = 0.024, respectively) and increased time to recurrence (TTR) (p = 0.037 and p = 0.031, respectively). In addition, combined analysis of CD86 and CD206 provided a better indicator for OS (p = 0.011) and TTR (p = 0.024) in HCC than individual analysis of CD86 and CD206. Moreover, CD86⁺/CD206⁺ TAMs predictive model also had significant prognosis value in α-fetoprotein (AFP)-negative patients (OS: p = 0.002, TTR: p = 0.005). Thus, these results suggest that combined analysis of immune biomarkers CD86 and CD206 could be a promising HCC prognostic biomarker.


Interfacial growth of large-area single-layer metal-organic framework nanosheets.
The air/liquid interface is an excellent platform to assemble two-dimensional (2D) sheets of materials by enhancing spontaneous organizational features of the building components and encouraging large length scale in-plane growth. We have grown 2D molecularly-thin crystalline metal-organic-framework (MOF) nanosheets composed of porphyrin building units and metal-ion joints (NAFS-13) under operationally simple ambient conditions at the air/liquid interface. In-situ synchrotron X-ray diffraction studies of the formation process performed directly at the interface were employed to optimize the NAFS-13 growth protocol leading to the development of a post-injection method -post-injection of the metal connectors into the water subphase on whose surface the molecular building blocks are pre-oriented- which allowed us to achieve the formation of large-surface area morphologically-uniform preferentially-oriented single-layer nanosheets. The growth of such large-size high-quality sheets is of interest for the understanding of the fundamental physical/chemical properties associated with ultra-thin sheet-shaped materials and the realization of their use in applications.


Protective role of excitatory amino acid antagonists in experimental retinal ischemia.
Excitatory amino acids and their analogues (NMDA, kainate and AMPA) are implicated in the pathogenesis of ischemic brain injury. In order to fully understand their involvement in the pathogenesis of retinal ischemic injury, we studied the electrophysiological and histopathological effects of two excitatory amino acid antagonists, cis-PDA and MK 801, in an experimental retinal ischemia model. The two antagonists were injected intravitreously 15 min before ischemia was induced by elevatory intraocular pressure caused by external compression. Electrophysiological and histopathological evaluation was made 48 h after 45 min transient ischemia. The excitatory amino acid antagonists cis-PDA and MK 801 can partially protect against retinal ischemic injury; whereas the mean post-ischemic b-wave amplitude corresponded to 41% of the pre-ischemic value in the control group, it was 64% (P = 0.003) and 59% (P = 0.005) following administration of cis-PDA and MK 801 respectively. Histopathological study corroborated these data, showing significant differences for morphometric parameters (P = 0.011 and P = 0.007 respectively). These preliminary results suggest the possibility of limiting excito-toxicity, one of the lesion-forming mechanisms in ischemic retinal injury.


FUNYBASE: a FUNgal phYlogenomic dataBASE.
The increasing availability of fungal genome sequences provides large numbers of proteins for evolutionary and phylogenetic analyses. However the heterogeneity of data, including the quality of genome annotation and the difficulty of retrieving true orthologs, makes such investigations challenging. The aim of this study was to provide a reliable and integrated resource of orthologous gene families to perform comparative and phylogenetic analyses in fungi. FUNYBASE is a database dedicated to the analysis of fungal single-copy genes extracted from available fungal genomes sequences, their classification into reliable clusters of orthologs, and the assessment of their informative value for phylogenetic reconstruction based on amino acid sequences. The current release of FUNYBASE contains two types of protein data: (i) a complete set of protein sequences extracted from 30 public fungal genomes and classified into clusters of orthologs using a robust automated procedure, and (ii) a subset of 246 reliable ortholog clusters present as single copy genes in 21 fungal genomes. For each of these 246 ortholog clusters, phylogenetic trees were reconstructed based on their amino acid sequences. To assess the informative value of each ortholog cluster, each was compared to a reference species tree constructed using a concatenation of roughly half of the 246 sequences that are best approximated by the WAG evolutionary model. The orthologs were classified according to a topological score, which measures their ability to recover the same topology as the reference species tree. The full results of these analyses are available on-line with a user-friendly interface that allows for searches to be performed by species name, the ortholog cluster, various keywords, or using the BLAST algorithm. Examples of fruitful utilization of FUNYBASE for investigation of fungal phylogenetics are also presented. FUNYBASE constitutes a novel and useful resource for two types of analyses: (i) comparative studies can be greatly facilitated by reliable clusters of orthologs across sets of user-defined fungal genomes, and (ii) phylogenetic reconstruction can be improved by identifying genes with the highest informative value at the desired taxonomic level.


In Patients with Grade I and II Ankle Sprains, Dynamic Taping Seems to Be Helpful during Certain Tasks, Exercises and Tests in Selected Phases of the Rehabilitation Process: A Preliminary Report.
We aimed to investigate changes in postural stability on a stable surface after the application of dynamic tape for patients with inversion ankle sprains. This study enrolled 30 patients (age 25.5 ± 8.0 years) with grade I and II ankle sprains, which occurred 7−21 days before enrolment. Postural stability (balance, coordination, feedback) was assessed before and after the application of dynamic tape using a stabilographic platform. Three 32-s exercises were performed on the stabilographic platform, one with eyes open, one with eyes closed and one with visual feedback. After the application of dynamic tape, an improvement was observed in terms of the mean radius of sway (4.2 ± 1.3 mm vs. 3.4 ± 0.9 mm; p = 0.012) and coordination (48.8 ± 19.2% vs. 59.3 ± 5.8%; p = 0.021). Selected balance parameters did not improve significantly in the tests with open and closed eyes. Asymmetric load improved for all tests, but significant differences were only observed with eyes closed (34.9 ± 24.4 vs. 41.7 ± 30.5; p < 0.01). We concluded that the use of dynamic tape after an ankle sprain significantly improved balance and coordination on a stable surface. The benefits were shown in terms of a significant improvement in the asymmetric load of the injured limb in comparison to the healthy limb during the test with closed eyes and a considerable improvement in the asymmetric load that was evaluated with visual feedback on a stable surface.


The pea late nodulin gene PsNOD6 is homologous to the early nodulin genes PsENOD3/14 and is expressed after the leghaemoglobin genes.
The pea late nodulin gene PsNOD6 has been cloned and sequenced. PsNOD6 is homologous to the pea early nodulin genes PsNOD3 and PsENOD14. In situ hybridization experiments showed that, like the PsENOD3 and PsENOD14 genes, the PsNOD6 gene is only expressed in the infected cell type. The PsNOD6 gene is first expressed at the transition of the pre-fixation zone II into the interzone II-III (the amyloplast-rich zone preceding the fixation zone III), whereas the early nodulin genes PsENOD3 and PsENOD14 are already induced in the pre-fixation zone II. Thus these nodulin genes encoding homologous proteins are induced at consecutive stages of nodule development. The expression of the late nodulin genes encoding leghaemoglobin precedes the expression of the late nodulin gene PsNOD6. Therefore these late nodulin genes have to be regulated by different mechanisms despite the fact they are expressed in the same cell type. This conclusion is consistent with the fact that PsNOD6 lacks one of the conserved regions occurring in the promoters of all other late nodulin genes studied.


Cytogenetic effects of styrene and styrene oxide.
Styrene and styrene oxide induce various cytogenetic effects, similar in both human lymphocytes in vitro and onion root-tip cells in vivo. Styrene appears to cause chromosome breakage in both systems, and in Allium it shows a strong c-mitotic effect. Styrene oxide, on the other hand, seems to destroy the tertiary folding of the chromatin. Cytotoxicity of styrene oxide is very high (complete mitotic inhibition occurs on 0.03% v/v) in human lymphocytes, whereas, in Allium, styrene is slightly more toxic than styrene oxide. Styrene glycol, a further metabolite of styrene oxide, does not cause mitotic inhibition.


Lack of increase in concentrations of cerebrospinal fluid sodium in rats with various stages of DOCA-salt hypertension.
Experiments were conducted in conscious rats to determine whether DOCA-salt treatment could cause an elevation of sodium concentration of cerebrospinal fluid (CSF), which may be responsible for the enhanced activity of sympathetic nervous system (SNS) and increased secretion of vasopressin (AVP). Systolic blood pressure (SBP) and mean arterial pressure (MAP) were gradually but consistently increased by DOCA-salt treatment. Serum Na concentration was similarly increased with time by DOCA-salt, and significantly higher than control in the 4th treatment week. In contrast, DOCA-salt did not alter the CSF Na levels at any time during treatment. A relationship between SBP and CSF Na was never evident at any stage of the DOCA-salt hypertension. The decrease in MAP following administration of the vasopressin V1-receptor antagonist, d(CH2)5Tyr(Me)AVP (30 micrograms/kg), or hexamethonium (30 mg/kg) was enhanced in the DOCA-treated rats, as compared to findings in the controls. These hypotensive effects were gradually, but progressively enhanced with the development of hypertension by DOCA-salt treatment. We tentatively conclude that mechanisms accounting for the enhanced activity of SNS and AVP in DOCA-salt hypertensive rats are independent of an increased Na concentration in the CSF.


Phosphate removal from synthetic and real wastewater using steel slags produced in Europe.
Electric arc furnace steel slags (EAF-slags) and basic oxygen furnace steel slags (BOF-slags) were used to remove phosphate from synthetic solutions and real wastewater. The main objective of this study was to establish an overview of the phosphate removal capacities of steel slags produced in Europe. The influences of parameters, including pH, and initial phosphate and calcium concentrations, on phosphate removal were studied in a series of batch experiments. Phosphate removal mechanisms were also investigated via an in-depth study. The maximum capacities of phosphate removal from synthetic solutions ranged from 0.13 to 0.28 mg P/g using EAF-slags and from 1.14 to 2.49 mg P/g using BOF-slags. Phosphate removal occurred predominantly via the precipitation of Ca-phosphate complexes (most probably hydroxyapatite) according to two consecutive reactive phases: first, dissolution of CaO-slag produced an increase in Ca(2+) and OH(-) ion concentrations; then the Ca(2+) and OH(-) ions reacted with the phosphates to form hydroxyapatite. It was found that the release of Ca(2+) from slag was not always enough to enable hydroxyapatite precipitation. However, our results indicated that the Ca(2+) content of wastewater represented a further source of Ca(2+) ions that were available for hydroxyapatite precipitation, thus leading to an increase in phosphate removal efficiencies.


Epistatic gene interactions in the control of division in fission yeast.
THERE is currently much interest in the mechanism which controls the timing of cell division. Certain features of the control have been found to be common to a variety of eukaryotes. In particular, the importance of cell size as a parameter affecting cell cycle progress has been reported for mammalian cells(1,2) and for several single-celled eukaryotes(3-6). Another feature common to several systems is that growth conditions have a direct effect on the timing of division cycle events(7-9), and on cell size(9,10). In the fission yeast Schizosaccharomyces pombe, both cell size(6) and nutritional conditions(9) have been shown to affect cycle kinetics. The organism has been used extensively as a model eukaryotic system, largely because of the ease of measuring cell size and because division occurs by binary fission(11). More recently, its genetic tractability has led to the isolation of cell division cycle (cdc) mutants(12), and also of wee mutants altered in the control coordinating growth with the division cycle(13-15). The existence of such control mutants allows a more direct approach to the investigation of the molecular basis of division control, in contrast to the indirect methods used in other systems(4,16-18). wee mutants are so far unique to S. pombe. The most conspicuous property of wee mutants is their reduced cell size(13,14). Analysis of these mutants(15,19) and other evidence(9) has shown that control over cell division timing normally acts at entry to mitosis. As the function of a number of cdc genes is specifically required for mitosis(12), interactions between wee and cdc mutants which affect mitosis might be expected. I report here that the mitotic defect caused by a defective cdc25 allele is suppressed in wee mutants. Suppression by wee1 mutants is almost complete, while the wee2.1 mutation is a less effective suppressor. The significance of these findings for genetic models of the control of mitosis is considered.


Childhood bullous pemphigoid: report of three cases.
Bullous pemphigoid (BP) is a disorder that rarely occurs in children. We hereby describe three cases of childhood BP aged 2-4 months, which are among the youngest reported in the literature. BP was confirmed by histopathology, direct and indirect immunofluorescence with salt-split skin test and immunoblotting. These patients were successfully treated with systemic corticosteroids with a complete clinical remission.


The true cost of cardiovascular imaging: focusing on downstream, indirect, and environmental costs.
To develop a more realistic assessment of costs, herein named "true" costs, the extra-cancer from medical radiation, environmental damage from imaging paraphernalia and radioactive wastes must be included as long-term costs from imaging examinations. It is urgent to define the "true" costs across imaging modalities as it interferes on physicians' decision to request an exam and on research projects such as cost-effectiveness analysis. Cardiology is the specialty that most will benefit from the outcome as cardiovascular exams represent almost 30% of the total exams acquired annually worldwide.


Rotator cuff disorders.
Rotator cuff problems present with shoulder pain on repetitive overhead activity. Chronic irritation may develop into impingement tendonitis, with weakness of abduction and external rotation and night pain. Conservative management with rest, anti-inflammatory medicine and physiotherapy resolves the majority of symptoms. If these persist, surgical decompression affords good relief of pain.


Sex hormones impact early maturation and immune response in the arteriovenous fistula mouse model.
Arteriovenous fistulae (AVF) fail to mature more frequently in female patients compared with male patients, leading to inferior outcomes and decreased utilization. Since our mouse AVF model recapitulates sex differences in human AVF maturation, we hypothesized that sex hormones mediate these differences during AVF maturation. C57BL/6 mice (9-11 wk) were treated with aortocaval AVF surgery and/or gonadectomy. AVF hemodynamics were measured via ultrasound (days 0-21). Blood was collected for FACS and tissue for immunofluorescence and ELISA (days 3 and 7); wall thickness was assessed by histology (day 21). Inferior vena cava shear stress was higher in male mice (P = 0.0028) after gonadectomy, and they had increased wall thickness (22.0 ± 1.8 vs. 12.7 ± 1.2 µm; P < 0.0001). Conversely, female mice had decreased wall thickness (6.8 ± 0.6 vs. 15.3 ± 0.9 µm; P = 0.0002). Intact female mice had higher proportions of circulating CD3+ T cells on day 3 (P = 0.0043), CD4+ (P = 0.0003) and CD8+ T cells (P = 0.005) on day 7, and CD11b+ monocytes on day 3 (P = 0.0046). After gonadectomy, these differences disappeared. In intact female mice, CD3+ T cells (P = 0.025), CD4+ T cells (P = 0.0178), CD8+ T cells (P = 0.0571), and CD68+ macrophages (P = 0.0078) increased in the fistula wall on days 3 and 7. This disappeared after gonadectomy. Furthermore, female mice had higher IL-10 (P = 0.0217) and TNF-α (P = 0.0417) levels in their AVF walls than male mice. Sex hormones mediate AVF maturation, suggesting that hormone receptor signaling may be a target to improve AVF maturation.NEW & NOTEWORTHY After arteriovenous fistula creation, females have lower rates of maturation and higher rates of failure than males. In a mouse model of venous adaptation that recapitulates human fistula maturation, sex hormones may be mechanisms of the sexual dimorphism: testosterone is associated with reduced shear stress, whereas estrogen is associated with increased immune cell recruitment. Modulating sex hormones or downstream effectors suggests sex-specific therapies and could address disparities in sex differences in clinical outcomes.


Gene expression profiling of primary tumor cell populations using laser capture microdissection, RNA transcript amplification, and GeneChip microarrays.
Gene expression profiling from microdissected cell populations is a powerful approach to explore molecular processes involved in development and solid tumor biology. In this chapter, we detail robust and validated methods for tissue preparation and isolation of high-quality RNA from microdissected cell populations. A protocol is also provided for linear transcript amplification using as little as 10 ng of total RNA to produce labeled cRNA targets for hybridization to GeneChip high-density oligonucleotide microarrays. Particular emphasis is placed on troubleshooting each technical step in the protocol and measures of quality assurance for both RNA isolation and resulting microarray data.


Localization of human serum amyloid P component and heparan sulfate proteoglycan in in vitro-formed Abeta fibrils.
Ultrastructural studies of the localization of serum amyloid P component (SAP) in amyloid fibrils have given divergent results. We here report for the first time that electron microscopy of SAP coincubated with Abeta1-42 peptides or with mature Abeta1-42 fibrils, revealed SAP molecules coating the surface of the mature fibrils and that protofibrils of Abeta1-42 did not bind SAP. Also when incubated with extracted amyloid light chain (AL)-fibrils the SAP molecules aligned on the fibril surface. Heparan sulfate proteoglycan bound to the surface of the Abeta fibrils with a spacing of about 50 nm. We conclude that SAP does not bind to protofibrils but to the surface of mature Abeta fibrils and that it may stabilize and protect the fibrils.


Contraction of urinary bladder by central norepinephrine originating in the locus coeruleus.
Studies were performed to elucidate the role of the locus coeruleus, which is rich in norepinephrine-containing cell bodies, in vesical function using alpha-chloralose anesthetized cats. Stimulation of the locus coeruleus caused contraction of the urinary bladder, which was not affected by transection of the bilateral hypogastric nerves, but blocked by intravenous administration of hexamethonium, a ganglion blocking agent. In animals with transected hypogastric nerves, the locus coeruleus-induced contraction was inhibited by intrathecal administration of phentolamine (alpha-blocker) and prazosin (alpha 1-blocker), but not affected by intrathecal sotalol (beta-blocker) or yohimbine (alpha 2-blocker). In animals treated with reserpine, the locus coeruleus-induced contraction was enhanced by intravenous application of L-dopa, a precursor of norepinephrine. These results suggest that norepinephrine derived from the locus coeruleus activated preganglionic neurons in the sacral intermediolateral nuclei via alpha 1-receptors, thereby producing urinary bladder contraction.


In vitro and in vivo activity of antimicrobial peptides synthesized based on the insect defensin.
Synthetic antimicrobial 9-mer peptides were designed from the amino acid sequence of an active site of insect defensin to increase the number of positively charged amino acid residues. These peptides, RLRLRIGRR-NH2, RLLLRIGRR-NH2 and RLYLRIGRR-NH2, showed strong antimicrobial activity against bacteria and fungus. These peptides showed no growth inhibition activity against murine fibroblasts or macrophages and no hemolytic activity against rabbit erythrocytes in vitro. Furthermore, the administration of these peptides protected mice from a lethal methicillin-resistant Staphylococcus aureus (MRSA) challenge. In addition, these peptides suppressed tumor necrosis factor alpha (TNF-alpha) gene expression and production induced by lipopolysaccharide (LPS) or lipoteichoic acid (LTA) in murine macrophages.


Outcomes of microincision cataract surgery versus coaxial phacoemulsification.
To compare outcomes of microincision cataract surgery (MICS) with coaxial phacoemulsification. Prospective randomized consecutive case series. One hundred eyes of 50 patients with nuclear or corticonuclear cataract (grades 2+ to 4) with Lens Opacities Classification System III. One hundred eyes (50 patients) were randomly operated through clear corneal incisions using 2 techniques: coaxial phacoemulsification (50 eyes) and microincision cataract surgery (50 eyes). Mean phacoemulsification time, total phacoemulsification percent, effective phacoemulsification time (EPT) (calculated by multiplying total phacoemulsification time in seconds by the average power percent used), intraoperative total balanced salt solution (BSS) volume, visual outcome, vectorial astigmatic changes, corneal thickness, endothelial cell count, and anterior chamber flare and cells preoperatively and at 1 day, 1 month, and 3 months. Statistically significant differences were found between MICS and coaxial phacoemulsification regarding mean incision size, mean total phacoemulsification percent, and EPT. There were no significant differences between the techniques regarding the mean percent of endothelial cell loss, anterior chamber cell count and flare, mean phacoemulsification time, pachymetric measures or total BSS volume utilized, or visual outcome. The vectorial astigmatic changes in the MICS group showed a change of < or =0.25 diopters (D) in 35% of the eyes, 0.25 to 0.5 D in 50% of the eyes, and 0.5 to 1.0 D in 15% of the eyes. These changes were induced by the surgery. Vectorial astigmatic changes of >1 D were not observed. In the coaxial phacoemulsification group, vectorial astigmatic changes of <0.25 D were not observed either. Changes of 0.25 to 0.5 D were seen in 20% of the eyes, and changes of 0.5 to 1.0 D were seen in 30%. Fifty percent of the eyes showed changes of >1.0 D. Mean vectorial astigmatic changes were 0.36+/-0.23 D in the MICS group and 1.2+/-0.74 D in the coaxial phacoemulsification group (P<0.001). Microincision cataract surgery significantly lowered mean phacoemulsification time, mean total phacoemulsification percent, mean EPT, and surgically induced astigmatism when compared with coaxial phacoemulsification.


Persistent Lin28 Expression Impairs Neurite Outgrowth and Cognitive Function in the Developing Mouse Neocortex.
Many neurodevelopmental disorders feature learning and memory difficulties. Regulation of neurite outgrowth during development is critical for neural plasticity and memory function. Here, we show a novel regulator of neurite outgrowth during cortical neurogenesis, Lin28, which is an RNA-binding protein. Persistent Lin28 upregulation by in utero electroporation at E14.5 resulted in neurite underdevelopment during cortical neurogenesis. We also showed that Lin28-overexpressing cells had an attenuated response to excitatory inputs and altered membrane properties including higher input resistance, slower action potential repolarization, and smaller hyperpolarization-activated cation currents, supporting impaired neuronal functionality in Lin28-electroporated mice. When we ameliorated perturbed Lin28 expression by siRNA, Lin28-induced neurite underdevelopment was rescued with reduction of Lin28-downstream molecules, high mobility group AT-Hook 2, and insulin-like growth factor 1 receptor. Finally, Lin28-electroporated mice showed significant memory deficits as assessed by the Morris water maze test. Taken together, these findings demonstrate a new role and the essential requirement of Lin28 in developmental control of neurite outgrowth, which has an impact on synaptic plasticity and spatial memory. These findings suggest that targeting Lin28 may attenuate intellectual disabilities by correction of impaired dendritic complexity, providing a novel therapeutic candidate for treating neurodevelopmental disorders.


Differential effect of the ultra-low dose and standard estrogen plus dydrogesterone therapy on thrombin generation and fibrinolysis in postmenopausal women.
The objective of this study was to estimate the effects of different doses of oral hormone therapy (HT) on thrombin generation and fibrinolytic activity in postmenopausal women after 12 months of treatment. Thrombin generation, fibrinolysis activators and inhibitors were determined before and after 12 months of treatment. Participants (180) were assigned (1:1:1) as follows: (1) standard HT group, 17β-estradiol (1 mg/day) with dydrogesterone (5 mg/day); (2) ultra-low dose HT group, 17β-estradiol (0.5 mg/day) with dydrogesterone (2.5 mg/day); (3) control group, no treatment. The standard HT led to a higher concentration of prothrombin 1 + 2 fragment (by 5.8%) with lower antithrombin activity (by 6.1%). Compared with baseline, we observed a reduction in mean antithrombin activity in the standard HT group and increases in mean levels of prothrombin 1 + 2 fragment in two HT groups. We found decreases after treatment in both standard and ultra-low dose HT groups in plasminogen activator inhibitor-1 (PAI-1) activity (-32.4% and -19.6%, respectively) and PAI-1 antigen (-9.9% and -7.8%, respectively). Intergroup analysis revealed reduction in both mean PAI-1 activities and PAI-1 antigen levels in the two treatment groups when compared with the control. Contrary to the standard estrogen plus dydrogesterone treatment, ultra-low dose HT revealed positive effects on hemostasis by intensifying fibrinolysis through a decrease in both PAI-1 activity and antigen levels, and with no impact on thrombin generation.


The normal scalp topography of the middle latency auditory evoked potential Pa component following monaural click stimulation.
The scalp topography of the middle latency auditory evoked potential (MLAEP) Pa component following left and right ear click stimulation was investigated in 15 normal hearing and neurologically intact right-handed subjects. An unbiased reference was employed. The Pa component showed a broad voltage field that was recorded maximally at the Cz and Fz leads regardless of which ear was stimulated. A broad negative voltage field that occurred coincident in time with the Pa component was recorded posterior to the T3, P3, Pz, P4 and T4 electrode leads. This negative voltage field peaked in amplitude at the T5, O1, Oz, O2, and T6 electrode leads. An unexpected finding was that the peak latency of the Pa component occurred significantly earlier following stimulation of the right ear.


Reacting ultrastructure of the human oligodendrocyte (a study of cerebral cortex distant to brain tumours).
Electron microscopical examination of adjacent human cerebral cortex from patients with brain tumour revealed the following morphological distinctive features of the oligodendrocyte: the presence of a diffuse electron dense material, large quantities of free ribosomes or ribosomal rosettes, eccentric nucleus with irregular clumps of chromatin, swollen perinuclear cisterna, extensive cytoplasmic microtubules, well-developed Golgi apparatus opposite the eccentric nuclei, dilated endoplasmic reticulum and mitochondria. The majority of oligodendrocytes contain large indefinable heterogeneous electron-dense structures within their perikaryon or processes. Oligodendrocytes with similar inclusion bodies are seen in the vicinity of capillaries. For these reasons a possible phagocytic activity of the human oligodendrocyte seems likely.


A polysaccharide from Dictyophora indusiata inhibits the immunosuppressive function of cancer-associated fibroblasts.
Reversing the function of cancer-associated fibroblasts (CAFs) may improve the efficacy of cancer therapy. Here, we isolated a novel polysaccharide from Dictyophora indusiata (ZSP4) and examined its effects on the function of prostate CAFs. The supernatant of prostate CAFs can stimulate the proliferation of immune cells and inhibit the growth of CD4+/CD8+ T cells. However, after ZSP4 stimulation, the functions of prostate CAFs were inhibited. The mechanism experiment shows that ZSP4 can stimulate prostate CAFs by down-regulating the expression of α-smooth muscle actin. Polysaccharides extracted from Dictyophora indusiata stimulate the proliferation of immune cells and reverse the immune-suppressive functions of prostate CAFs, shedding new light on the development of novel anticancer strategies. The endocrine therapy used to treat prostate cancer aims to eliminate androgenic activity from prostatic tissue; these therapies are painful and of poor therapeutic effect. In this study, we found that polysaccharides extracted from Dictyophora indusiata may affect the micro-environment of tumours and inhibit the growth of the tumours. Our results suggest that polysaccharides may modulate negative immune regulation and enhance antitumour immunity, which is important for clinical therapy.


Fibrin protofibril packing and clot stability are enhanced by extended knob-hole interactions and catch-slip bonds.
Fibrin polymerization involves thrombin-mediated exposure of knobs on one monomer that bind to holes available on another, leading to the formation of fibers. In silico evidence has suggested that the classical A:a knob-hole interaction is enhanced by surrounding residues not directly involved in the binding pocket of hole a, via noncovalent interactions with knob A. We assessed the importance of extended knob-hole interactions by performing biochemical, biophysical, and in silico modeling studies on recombinant human fibrinogen variants with mutations at residues responsible for the extended interactions. Three single fibrinogen variants, γD297N, γE323Q, and γK356Q, and a triple variant γDEK (γD297N/γE323Q/γK356Q) were produced in a CHO (Chinese Hamster Ovary) cell expression system. Longitudinal protofibril growth probed by atomic force microscopy was disrupted for γD297N and enhanced for the γK356Q mutation. Initial polymerization rates were reduced for all variants in turbidimetric studies. Laser scanning confocal microscopy showed that γDEK and γE323Q produced denser clots, whereas γD297N and γK356Q were similar to wild type. Scanning electron microscopy and light scattering studies showed that fiber thickness and protofibril packing of the fibers were reduced for all variants. Clot viscoelastic analysis showed that only γDEK was more readily deformable. In silico modeling suggested that most variants displayed only slip-bond dissociation kinetics compared with biphasic catch-slip kinetics characteristics of wild type. These data provide new evidence for the role of extended interactions in supporting the classical knob-hole bonds involving catch-slip behavior in fibrin formation, clot structure, and clot mechanics.


Physiological responses to fire fighting activities.
Eight professional fire fighters participated in six fire fighting scenarios at a training facility. Data on heart rate (HR), rectal temperature (Tre), and skin temperatures at the chest and thigh were collected using a portable data acquisition system. Average HR ranged from 122 to 151 beats.min-1 during the six scenarios. Detailed analyses indicated that HR and Tre increases are related to both the physical and environmental stresses of the various activities carried out. The most demanding activity, that of building search and victim rescue, resulted in an average HR of 153 beats.min-1 and Tre rise of 1.3 degree C, while the least demanding activity, that of the crew captain who directs the fire fighting, resulted in an average HR of only 122 beats.min-1 and a Tre rise of only 0.3 degree C. This study shows that fire fighting is strenuous work for those directly entering a building and performing related duties, but that the physical demands of other activities are considerably less. The results further suggest that heat strain injuries in fire fighters could perhaps be reduced by rotating duties frequently with other crew members performing less stressful work.


Does the family affluence scale reflect actual parental earned income, level of education and occupational status? A validation study using register data in Sweden.
To examine the external validity of the Family Affluence Scale (FAS) among adolescents in Sweden by using register data for parental earned income, level of education and occupational status. Data from the baseline (2015-2019) of the Study of Adolescence Resilience and Stress (STARS), comprising 2283 13-year-olds in the region of Västra Götaland, were used. The FAS III consists of six items: unshared bedroom, car ownership, computer/tablet ownership, dishwasher, number of bathrooms and number of holidays abroad. Register data regarding earned income, educational level and occupational status from Statistics Sweden (2014-2018) were linked to adolescents. In total, survey data were available for 2280 adolescents, and register data were available for 2258 mothers and 2204 fathers. Total parental earned income was moderately correlated with adolescents' scoring on FAS (0.31 < r < 0.48, p < 0.001), depending on examination year. The low FAS group mainly comprised low-income households, and the high FAS group mainly comprised high-income households. Correlations between mothers' and fathers' educational level and adolescents' scoring on FAS were low (r = 0.19 and r = 0.21, respectively, p < 0.001). FAS was higher among adolescents whose parents were working, but the correlation between parents' occupational status and FAS was low (r = 0.22, p < 0.001). The FAS can mainly identify low- and high-income households in Sweden. It may be used as an alternative measure of parental earned income in studies using self-reported socioeconomic status among adolescents.


Recent Treatment Advances in Amblyopia.
Occlusion therapy has a long history as the gold standard treatment for amblyopia. Over the past two decades, large multicenter randomized controlled trials and objective dose-monitoring studies have characterized the effects of refractive correction, patching, and atropine penalization, providing insights into the impact of factors such as age and treatment dose. More recent approaches, whose development has been accelerated by advances in technology, are designed to provide different stimulation to the amblyopic eye and the fellow eye. This review explores a variety of such dichoptic approaches, categorized according to whether they primarily feature requisite use of the amblyopic eye in the face of fellow-eye masking, integration of visual information from both eyes, or reduction of stimulus salience in the fellow eye. It is still unclear whether dichoptic treatments are superior to traditional, low-cost treatment methods or whether their therapeutic mechanisms are fundamentally different from those of established treatments.


Effects of Cathepsin S Inhibition in the Age-Related Dry Eye Phenotype.
Aged C57BL/6J (B6) mice have increased levels of cathepsin S, and aged cathepsin S (Ctss-/-) knockout mice are resistant to age-related dry eye. This study investigated the effects of cathepsin S inhibition on age-related dry eye disease. Female B6 mice aged 15.5 to 17 months were randomized to receive a medicated diet formulated by mixing the RO5461111 cathepsin S inhibitor or a standard diet for at least 12 weeks. Cornea mechanosensitivity was measured with a Cochet-Bonnet esthesiometer. Ocular draining lymph nodes and lacrimal glands (LGs) were excised and prepared for histology or assayed by flow cytometry to quantify infiltrating immune cells. The inflammatory foci (>50 cells) were counted under a 10× microscope lens and quantified using the focus score. Goblet cell density was investigated in periodic acid-Schiff stained sections. Ctss-/- mice were compared to age-matched wild-type mice. Aged mice subjected to cathepsin S inhibition or Ctss-/- mice showed improved conjunctival goblet cell density and cornea mechanosensitivity. There was no change in total LG focus score in the diet or Ctss-/- mice, but there was a lower frequency of CD4+IFN-γ+ cell infiltration in the LGs. Furthermore, aged Ctss-/- LGs had an increase in T central memory, higher numbers of CD19+B220-, and fewer CD19+B220+ cells than wild-type LGs. Our results indicate that therapies aimed at decreasing cathepsin S can ameliorate age-related dry eye disease with a highly beneficial impact on the ocular surface. Further studies are needed to investigate the role of cathepsin S during aging.


Voriconazole inhibits cross-kingdom interactions between Candida albicans and Actinomyces viscosus through the ergosterol pathway.
Candida albicans and Actinomyces viscosus are prominent microbes associated with dental root caries. The aim of this study was to investigate the effect of C. albicans on A. viscosus biofilms and to identify the mechanisms associated with this interaction. A. viscosus and C. albicans strains (wide-type and mutants) were used to form biofilms in vitro and in vivo, which were subsequently analysed by crystal violet assay and scanning electron microscopy (SEM) to investigate the effect of C. albicans on A. viscosus growth. A viable plate count and survival curve for C. albicans mutants and A. viscosus combinations were used to identify which C. albicans pathway was crucial for cross-kingdom interactions. Voriconazole was used to block their interactions both in vitro and in vivo. SEM, fluorescence in situ hybridisation (FISH), quantitative PCR and survival curve analyses were performed to evaluate the activity of voriconazole on C. albicans and A. viscosus interactions. The biomass and virulence of mixed-species biofilms were significantly enhanced compared with the A. viscosus biofilm alone. However, this was not observed in the mixed-species biofilms with the C. albicans mutant erg11Δ/Δ in vitro and in vivo, indicating that azoles may work on the mixed-species biofilms. As expected, voriconazole can effectively reduce the biomass of mixed-species biofilms. A high concentration of voriconazole (1 µg/mL) reduced the abundance of C. albicans, whilst a low voriconazole concentration (0.25 µg/mL) blocked their interactions similar to the effect of the erg11Δ/Δ mutant. Voriconazole may be a candidate strategy to combat root caries pathogens.


Influence of carrier materials and coatings on retinal pigment epithelium cultivation and functions.
Properties of retinal pigment epithelium (RPE) are relevant for the development of cell culture models concerning an exact reproduction of the ocular cell biology. Here, we want to investigate how different carrier materials and coatings influence proliferation, differentiation and functions of RPE in regard to development of a three-dimensional cell culture model based on primary porcine RPE. Human RPE cell line ARPE-19 and primary porcine RPE were used. Cells were cultivated on plates which were coated with collagen I, collagen IV, laminin or fibronectin, respectively, and cell numbers were assessed after different time periods via trypan blue staining. Also, the ARPE-19 were cultivated on polydimethylsiloxane (PDMS), alginate, gelatin methacrylate (GelMA), poly-N-isopropylacrylamide (PNIPAM) and cells number were assessed. Primary RPE were cultured on PDMS material. Supernatants were collected and analyzed via ELISA for their vascular endothelial growth factor (VEGF) and transforming growth factor β (TGF-β) content. After day 14 cells were lysed and retinal pigment epithelium-specific 65 kDa protein (RPE65) and bestrophin-1 (BEST1) expression was investigated via Western blot. Cellular functions were tested on collagen I, collagen IV, laminin and fibronectin with and without PDMS. Scratch assay was performed to detect wound healing 24 and 48 h after scratch application. Immunolabeling was used to highlight tight junctions in concert with Hoechst staining and phalloidin to label cell nuclei and actin filaments, respectively. Phagocytosis of fluorescently labeled latex beads opsonized with photoreceptor outer segments (POS) was assessed via fluorescence microscopy. Transepithelial electrical resistance was measured for detection of cellular barrier. Gene expression of RDH11 (retinol dehydrogenase 11), BEST1 (bestrophin 1) and TGFB1 (transforming growth factor beta 1) was investigated via real-time PCR. Only PDMS carrier material was appropriate for primary RPE and ARPE-19 cell cultivation. Coating of PDMS with laminin led to increased proliferation. In primary RPE, VEGF secretion was increased if PDMS was coated with laminin or fibronectin compared to uncoated PDMS. No significant changes in phagocytic ability and generation of tight junctions were detected between different coatings, but RPE65 expression was reduced on fibronectin coated PDMS. Laminin coating decreased TGF-β and increased BEST1 protein expression. Also, RPE on collagen IV showed highest TEER on transwell plates. The genes RDH11 and TGFB1 were decreased when coated with collagen IV without PDMS as well as coated PDMS. Laminin and collagen IV coating led to an increased wound healing. Cultivation of RPE and ARPE-1 on PDMS is a possible alternative for cell culture models whereas alginate, GelMA and PNIPAM were not suitable. Coating with laminin increased the proliferation, wound healing and VEGF secretion of the cells. The results suggest that laminin coated PDMS as carrier material is suitable for the development of 3D culture model systems.


Effects of vanillin, quillaja saponin, and essential oils on in vitro fermentation and protein-degrading microorganisms of the rumen.
This study investigated the effects of vanillin on methanogenesis and rumen fermentation, and the responses of ruminal protein-degrading bacteria to vanillin (at concentrations of 0, 0.76 and 1.52 g/L), essential oils (clove oil, 1 g/L; origanum oil, 0.50 g/L, and peppermint oil, 1 g/L), and quillaja saponin (at concentration of 0 and 6 g/L) in vitro. Methane production, degradabilities of feed substrate, and ammonia concentration decreased linearly with increasing doses of vanillin. Concentration of total volatile fatty acids also decreased, whereas proportion of butyrate tended to increase linearly with increasing doses of vanillin. Protozoa population decreased, but abundances of Ruminococcus flavefaciens, Prevotella bryantii, Butyrivibrio fibrisolvens, Prevotella ruminicola, Clostridium aminophilum, and Ruminobacter amylophilus increased with increasing doses of vanillin. Origanum and clove oils resulted in lower ammonia concentrations compared to control and peppermint oil. All the tested essential oils decreased abundances of protozoa, Selenomonas ruminantium, R. amylophilus, P. ruminicola and P. bryantii, with the largest decrease resulted from origanum oil followed by clove oil and peppermint oil. The abundances of Megasphaera elsdenii, C. aminophilum, and Clostridium sticklandii were deceased by origanum oil while that of B. fibrisolvens was lowered by both origanum and clove oils. Saponin decreased ammonia concentration and protozoal population, but increased the abundances of S. ruminantium, R. amylophilus, P. ruminicola, and P. bryantii, though the magnitude was small (less than one log unit). The results suggest that reduction of ammonia production by vanillin and saponin may not be caused by direct inhibition of major known proteolytic bacteria, and essential oils can have different inhibitory effects on different proteolytic bacteria, resulting in varying reduction in ammonia production.


Membrane actions of progestins at dopamine type 1-like and GABAA receptors involve downstream signal transduction pathways.
In the ventral tegmental area (VTA), progestins facilitate lordosis via rapid actions at membrane dopamine Type 1-like (D(1)) and/or GABA(A) receptors (GBRs), rather than via cognate, intracellular progestin receptors (PRs). Downstream signal transduction pathways involved in these effects were investigated using lordosis as a bioassay. If progestins' actions at D(1) and/or GBRs in the VTA require activation of G-proteins, adenylyl cyclase, cyclic AMP-dependent protein kinase A (PKA), phospholipase C (PLC), and/or PKC, then pharmacologically blocking these pathways would be expected to attenuate progestin-facilitated lordosis and its enhancement by D(1) and GBR activity. Ovariectomized, estradiol-primed rats were infused first with vehicle or signal transduction inhibitor, and second with vehicle, a D(1) or GBR agonist, and then with vehicle or progestins to the VTA. Rats were tested for lordosis following infusions. Results indicated that initiation of G-proteins, adenylyl cyclase, PKA, PLC, or PKC in the VTA is required for rapid effects of progestins through D(1) and/or GBRs to facilitate lordosis. As well, progestins' actions at n-methyl-d-aspartate receptors (NMDARs) may modulate activity at D(1) and/or GBRs and mitogen activated protein kinase (MAPK) may be a common signaling pathway. Findings from a microarray study demonstrated that there was upregulation of genes associated with steroid metabolism, GBRs, D(1), NMDARs and signal transduction factors in the midbrain VTA of naturally receptive mated compared to non-mated rats. Thus, in the VTA, progestins have rapid membrane-mediated actions via D(1), GBRs, NMDARs and their downstream signal transduction pathways.


Estimating the environmental impact of micro-pollutants in the low Ebro River (Spain): an approach based on screening toxicity with Vibrio fischeri.
The aim of this study was to assess the likely impacts on the ecosystems due to agricultural, human, and industrial activities carried out in an ecologically important area of the Ebro River (Spain). For it, a screening site specific ecological risk assessment was conducted. Considering the presence of high levels of potentially toxic substances, such as metals and chlorinated organic compounds, aqueous and organic extracts were used to assess toxicity in sediments by using the photo-luminescent bacteria Vibrio fischeri (Microtox) as screening response variable. Sediment samples collected during 2005-2006 in the last course of the Ebro River and its Delta have been analyzed. Toxic responses have shown strong relationships to the levels of pollutants in the area. Moreover, various sites presented some toxicity level, probably because of other factors associated with reducing environments into the sediments. Results indicate that Microtox bioassay is an appropriate tool to perform risk assessment studies at screening level.


Investigation of Reflux Characteristics in Outpatients of Otorhinolaryngology-Head and Neck Surgery by Age and Gender in the Chinese Population.
To investigate the characteristics of patients with reflux diseases in the otorhinolaryngology-head and neck surgery clinics through the Reflux Symptom Scale-12 (RSS-12) and the Gastroesophageal Reflux Disease Questionnaire (GERD-Q). All included patients completed the RSS-12 and GERD-Q scales and were considered to have LPR with an RSS-12 score >11 and GERD with a GERD-Q score >7. Data were analyzed according to genders (male and female) and age (18-40, 41-60, and >60 years). A total of 977 patients were included. the mean RSS-12 and GERD-Q score were 11.32±21.34 and 6.31±1.21, and the positive rate of LPR and GERD were 28.76% and 8.90%, respectively. Males had a higher positive rate of LPR and GERD than females, and there were more males with LPR who also had GERD. Among those with both LPR and GERD, males had significantly higher ear-nose-throat (ENT) symptom scores such as hoarseness and excess throat mucus than females. However, females had significantly higher scores of gastrointestinal (GI) symptoms, mainly indigestion and abdominal pain, and elder patients (>60 years) had higher scores of ENT, GI, respiratory symptoms, as well as the impact of symptoms on quality of life than the young patients (18-40, and 41-60 years). Patients in the otorhinolaryngology-head and neck surgery clinics have different reflux characteristics by gender and age in the Chinese population. Males had more severe ENT-related symptoms of distress, while females had more complaints of GI symptoms. Older patients had higher scores for ENT, GI and respiratory symptoms.


Elevated expression of secondary, but not early, responding genes to phorbol ester tumor promoters in papillomas and carcinomas of mouse skin.
A single topical treatment of mouse skin with the potent tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) results in transient inductions of a variety of genes. Based on the time courses of their inductions, these genes can be classified into two main groups: "early" response genes whose mRNA expression reaches a maximum 0.5-2 h after TPA treatment and "secondary" response genes whose mRNA expression is maximal 4 h or more after treatment. The nuclear oncogenes c-fos, c-myc, and c-jun belong to the early response group, whereas the metallothionein, osteopontin, and urokinase genes belong to the secondary response group. The steady-state expressions of these early and secondary response genes are all very low in normal skin, except that of c-jun, which is relatively high. Steady-state levels of expression and inducibility of these genes by TPA were not altered in initiated skin or in apparently normal skin during tumor promotion. We examined the expressions of these genes in papillomas and carcinomas produced by two-stage (initiator-promoter) and three-stage (initiator-promoter-initiator) protocols in mouse skin. Steady-state expression of the early responding nuclear oncogenes in papillomas and carcinomas was found to remain at the same low level as in normal skin. However, all the secondary responding genes were found to be expressed constitutively at high levels in these tumors. Elevated expressions of the genes for transforming growth factor alpha and beta were also observed in papillomas and to varying extents in carcinomas. These observations suggest that the regulatory machinery for transcription by the protein kinase C-mediated pathway through nuclear oncogenes is altered during the processes of tumor promotion and progression. The genes whose expression is elevated may be associated directly or indirectly with tumor promotion and progression.


Asthma and allergies: is the farming environment (still) protective in Poland? The GABRIEL Advanced Studies.
Evidence exists that a farming environment in childhood may provide protection against atopic respiratory disease. In the GABRIEL project based in Poland and Alpine regions of Germany, Austria and Switzerland, we aimed to assess whether a farming environment in childhood is protective against allergic diseases in Poland and whether specific exposures explain any protective effect. In rural Poland, 23 331 families of schoolchildren completed a questionnaire enquiring into farming practices and allergic diseases (Phase I). A subsample (n = 2586) participated in Phase II involving a more detailed questionnaire on specific farm exposures with objective measures of atopy. Farming differed between Poland and the Alpine centres; in the latter, cattle farming was prevalent, whereas in Poland 18% of village farms kept ≥1 cow and 34% kept ≥1 pig. Polish children in villages had lower prevalences of asthma and hay fever than children from towns, and in the Phase II population, farm children had a reduced risk of atopy measured by IgE (aOR = 0.72, 95% CI 0.57, 0.91) and skin prick test (aOR = 0.65, 95% CI 0.50, 0.86). Early-life contact with grain was inversely related to the risk of atopy measured by IgE (aOR = 0.66, 95% CI 0.47, 0.92) and appeared to explain part of the farming effect. While farming in Poland differed from that in the Alpine areas as did the exposure-response associations, we found in communities engaged in small-scale, mixed farming, there was a protective farming effect against objective measures of atopy potentially related to contact with grain or associated farm activities.


Effect of copper ions on transformation of organic sulfur in cationic exchange resins in Li2CO3-Na2CO3-K2CO3 molten-salt system.
Cationic exchange resins (CERs) were applied for purification and clarifying process of radioactive wastewater in nuclear industry, which was a kind of sulfur-containing organic material. Molten-salt oxidation (MSO) method can be applied for the treatment of spent CERs and the absorption of acid gas (such as SO2). The experiments about the molten salt destruction of the original resin and Cu ions doped resin were conducted. The transformation of organic sulfur in Cu ions doped resin was investigated. Compared with the original resin, the content of tail gas (such as CH4, C2H4, H2S and SO2) released from the decomposition of Cu ions doped resin was relatively high at 323-657 °C. Sulfur elements in the form of sulfates and copper sulfides were fixed in spent salt through XRD analysis. The XPS result revealed that the portion of functional sulfonic acid groups (-SO3H) in Cu ions doped resin was converted into sulfonyl bridges (-SO2-) at 325 °C. With the enhancement of temperature, sulfonyl bridges (-SO2-) were further decomposed to sulfoxides sulfur (-SO-) and organic sulfide sulfur. The destruction of thiophenic sulfur to H2S and CH4 was prompted by copper ions in copper sulfide. Sulfoxide were oxidized to the sulfone sulfur in molten salt. Sulfones sulfur consumed by reduction of Cu ions at 720 °C was more than it produced by oxidation of sulfoxide through XPS analysis, and the relative proportion of sulfone sulfur was 16.51%.


Detection of a nucleolar 7-2 ribonucleoprotein and a cytoplasmic 8-2 ribonucleoprotein with autoantibodies from patients with scleroderma.
In studies on antinucleolar antibodies in sera from 24 patients with scleroderma, an autoimmune disease, one serum, designated "anti-To", contained antibodies against a nucleolar 7-2 ribonucleoprotein and a novel cytoplasmic 8-2 ribonucleoprotein. The 7-2 and 8-2 RNAs are distinct RNAs with a pppG terminus. They are partially conserved between rat and human species and are present in distinct ribonucleoprotein particles. Eight sera contained antibodies that precipitated particles containing nucleolar U3 RNA; these antibodies appear to be directed against preribosomal particles containing U3 ribonucleoprotein, rather than the U3 ribonucleoprotein particles alone. All these ribonucleoproteins required proteins for antigenicity. These antibodies will be of use in studies on the structure and function of these novel small ribonucleoproteins.


Examining Barriers as Risk Factors for Relapse: A focus on the Canadian Treatment and Recovery System of Care.
In 2016, the Canadian Centre on Substance Use and Addiction (CCSA) conducted the first survey of individuals in recovery from addiction in Canada. The findings revealed that many individuals in recovery lead meaningful lives, contributing to their families and society. However, participants also identified a number of barriers to starting and maintaining recovery. The current study examined the relationship between the barriers experienced and relapse during recovery. Data from the 2016 Life in Recovery (LIR) from Addiction in Canada survey were analyzed using descriptive and logistic regression analyses. Participants comprised 855 individuals (Mage = 47.3 years), all of whom self-reported being in recovery from addiction. Logistic regressions revealed that upon starting recovery, long delays for treatment, odds ratio (OR) = 1.77, 95% confidence interval (CI) = 1.21-2.60, P < 0.01, and not having stable housing, OR = 1.83, 95% CI = 1.14-2.95, P < 0.05, were associated with increased risk of relapse. Moreover, upon examining barriers to maintaining recovery, a lack of supportive social networks, OR = 2.10, 95% CI = 1.26-3.48, p < 0.01, a lack of programs or supports, OR = 1.75, 95% CI = 1.03-2.98, P < 0.05, and the costs of recovery services OR = 1.73, 95% CI = 1.02-2.91, P < 0.05 were associated with increased risk of relapse. Targeted investments to address the treatment-related barriers that most strongly relate to relapse, could significantly improve the lives of individuals struggling with addiction and those beginning and maintaining their recovery journey. En 2016, le Centre canadien de lutte contre l'alcoolisme et les toxicomanies (CCLAT) a mené un premier sondage auprès des personnes en rétablissement à la suite d'une dépendance au Canada. Les résultats ont révélé que de nombreuses personnes en rétablissement mènent des vies significatives, contribuant ainsi à leur famille et à la société. Cependant, les participants ont également identifié un certain nombre d'obstacles au démarrage et au maintien du rétablissement. La présente étude a examiné la relation entre les obstacles rencontrés et la rechute pendant le rétablissement. Les données de l'enquête Life in Recovery (LIR) de l'Enquête sur la toxicomanie au Canada de 2016 ont été analysées à l'aide d'analyses de régression descriptives et logistiques. Les participants comprenaient 855 individus (Moyenne d'âge = 47,3 ans), qui ont tous déclaré être en rétablissement après une dépendance. Les régressions logistiques ont révélé qu'au début du rétablissement, de longs délais de traitement, chance de réussite (CR) = 1,77, intervalle de confiance à 95% (IC) = 1,21-2,60, p < 0,01, et n'ayant pas de logement stable, CR = 1,83, IC 95% = 1,14-2,95, p < 0,05, étaient associés à un risque accru de rechute. De plus, en examinant les obstacles au maintien du rétablissement, un manque de réseaux sociaux de soutien, CR = 2.10, IC à 95% = 1.26-3.48, p < 0.01, manque de programmes ou de soutien, CR = 1.75, IC à 95% = 1.03-2,98, p < 0,05, et les coÛts des services de récupération CR = 1,73, IC 95% = 1,02-2,91, p < 0,05 étaient associés à un risque accru de rechute. Des investissements ciblés visant à surmonter les obstacles liés au traitement les plus étroitement liés à la rechute pourraient améliorer considérablement la vie des personnes aux prises avec une dépendance et de celles qui amorcent et poursuivent leur chemin vers le rétablissement.


Sulfite sensitivity: relationship between sulfite plasma levels and bronchospasm: case report.
The present study describes a case of a 38-year-old nurse with asthma and probable sensitivity to sulfites. A double-blind challenge with sulfite and placebo was performed, pulmonary function tests were measured along with plasma levels of sulfites. A deficiency in sulfite-oxidase is suggested as the trigger mechanism of bronchospasm.


Treatment of chronic lymphocytic leukaemia and well-differentiated lymphocytic lymphoma with continuous low- or intermittent high-dose prednimustine versus chlorambucil/prednisolone.
Prednimustine, a new antitumour drug, is a chlorambucil ester of prednisolone. The present prospective randomized study compares the effect of continuous low-dose (B) and intermittent high-dose (C) prednimustine in previously untreated patients with progressive CLL and WDLL. The control group received continuous chlorambucil/prednisolone therapy (A). One hundred and eighteen patients, 88 CLL and 30 WDLL, were evaluable. Response to therapy (greater than 50% improvement) was noted in 61, 55 and 57% in groups A, B and C respectively. The difference was not statistically significant. Time to response, response duration and survival did not show any differences between the groups. Responding patients survived longer than patients with stationary and progressive disease. Median survival time was 72 months from diagnosis and 52 months from start of therapy, with no differences between the treatment groups. Toxicity of prednimustine was usually mild and similar to that of the two constituents. Treatment schedule C showed a slight advantage with regard to frequency of side effects. In conclusion, in this study the therapeutic effect of prednimustine was equal to that of its constituents administrated separately.


[Thoracic, abdominal and thoraco-abdominal traumas. A statistical-epidemiological study of 1009 cases and diagnostic-therapeutic considerations].
A study of 1009 cases of thoracic, abdominal and thoraco-abdominal traumas is presented. Relative complications are examined and some diagnostic and therapeutic considerations are put forward in relation to reported data.