Prolapsed valve of the foramen ovale in newborns and infants with coarctation of the aorta.
Out of 60 newborns and infants with symptomatic coarctation of the aorta, 34 had a significant left-to-right shunt at atrial level. In 20 of these 34 children, the atrial shunt had disappeared after surgical correction of the coarctation. All patients with this "reversible" shunt, thought to be secondary to an incompetent valve of the foramen ovale, were younger than 35 days at first cardiac catheterization. Left atrial pressure and left atrial volume of these patients did not differ significantly from those in a control group without atrial shunt. The conditions necessary for the development of a valve-incompetent foramen ovale include not only an increased left ventricular work load, but also a thin and pliable valve of the foramen ovale. It is only in infants younger than one month of age that the valve is thin and pliable enough to prolapse through the foramen ovale and form a channel between the two atria, which results in a left-to-right atrial shunt.


Microhomology-mediated End Joining and Homologous Recombination share the initial end resection step to repair DNA double-strand breaks in mammalian cells.
Microhomology-mediated end joining (MMEJ) is a major pathway for Ku-independent alternative nonhomologous end joining, which contributes to chromosomal translocations and telomere fusions, but the underlying mechanism of MMEJ in mammalian cells is not well understood. In this study, we demonstrated that, distinct from Ku-dependent classical nonhomologous end joining, MMEJ--even with very limited end resection--requires cyclin-dependent kinase activities and increases significantly when cells enter S phase. We also showed that MMEJ shares the initial end resection step with homologous recombination (HR) by requiring meiotic recombination 11 homolog A (Mre11) nuclease activity, which is needed for subsequent recruitment of Bloom syndrome protein (BLM) and exonuclease 1 (Exo1) to DNA double-strand breaks (DSBs) to promote extended end resection and HR. MMEJ does not require S139-phosphorylated histone H2AX (γ-H2AX), suggesting that initial end resection likely occurs at DSB ends. Using a MMEJ and HR competition repair substrate, we demonstrated that MMEJ with short end resection is used in mammalian cells at the level of 10-20% of HR when both HR and nonhomologous end joining are available. Furthermore, MMEJ is used to repair DSBs generated at collapsed replication forks. These studies suggest that MMEJ not only is a backup repair pathway in mammalian cells, but also has important physiological roles in repairing DSBs to maintain cell viability, especially under genomic stress.


[Change in the evoked potentials in the visual centers of the rat brain following administration of tetanus toxin to the lateral geniculate body].
Changes in visual evoked potentials were studied under conditions of tetanus-toxin injection into the lateral geniculate body in rats. The main abnormality of the evoked potentials in the geniculate body was the function of a new phase of the primary response. At the same time a strong increase in the amplitude of the first negative component of the evoked response in the ipsilateral cortical visual field was observed. The results show the formation of disturbances in the relay function of the lateral geniculate body during the formation of a generator of pathologically enhanced excitation in this nucleus.


In silico analysis and expression of a new chimeric antigen as a vaccine candidate against cutaneous leishmaniasis.
Since leishmaniasis is one of the health problems in many countries, the development of preventive vaccines against it is a top priority. Peptide vaccines may be a new way to fight the Leishmania infection. In this study, a silicon method was used to predict and analyze B and T cells to produce a vaccine against cutaneous leishmaniasis. Immunodominant epitope of Leishmania were selected from four TSA, LPG3, GP63, and Lmsti1 antigens and linked together using a flexible linker (SAPGTP). The antigenic and allergenic features, 2D and 3D structures, and physicochemical features of a chimeric protein were predicted. Finally, through bioinformatics methods, the mRNA structure was predicted and was produced chemically and cloned into the pLEXY-neo2 vector. Results indicated, polytope had no allergenic properties, but its antigenicity was estimated to be 0.92%. The amino acids numbers, molecular weight as well as negative and positive charge residuals were estimated 390, ~41KDa, 41, and 30, respectively. The results showed that the designed polytope has 50 post-translationally modified sites. Also, the secondary structure of the protein is composed of 25.38% alpha-helix, 12.31% extended strand, and 62.31% random coil. The results of SDS-PAGE and Western blotting revealed the recombinant protein with ~ 41 kDa. The results of Ramachandran plot showed that 96%, 2.7%, and 1.3% of amino acid residues were located in the preferred, permitted, and outlier areas, respectively. It is expected that the TLGL polytope will produce a cellular immune response. Therefore, the polytope could be a good candidate for an anti-leishmanial vaccine.


The long-term effect of specific type II 5alpha-reductase inhibition with finasteride on bone mineral density in men: results of a 4-year placebo controlled trial.
We determine the effect of long-term suppression of dihydrotestosterone with finasteride, a specific type II 5alpha-reductase inhibitor, on bone mineral density. As part of a large (3,040 cases) 4-year, double-blind, placebo controlled trial designed to assess the long-term effects of finasteride in men with benign prostatic hyperplasia, 157 men 46 to 76 years old who were randomized to receive either 5 mg. finasteride or placebo underwent dual energy x-ray absorptiometry of the lumbar spine at baseline and at years 2, 3 and 4. Of 117 patients who had a baseline measurement and at least 1 additional measurement during the study baseline mean plus or minus standard deviation bone mineral density values were 1.12 +/- 0.17 gm./cm.2 in the finasteride group (63) and 1.10 +/- 0.17 gm./cm.2 in the placebo group (54). After 4 years bone mineral density was not different between treatment groups (finasteride 1.14 +/- 0.17 gm./cm.2 and placebo 1.13 +/- 0.18 gm./cm.2). Similar results were obtained for the 33 finasteride and 25 placebo treated patients who completed the study with year 4 bone mineral density measurements. These data demonstrate that long-term inhibition of type II 5alpha-reductase with finasteride does not adversely affect bone mineral density.


Introduction of the Rapid Deployment Aortic Valve System Use in Elderly Patients With Endocarditis.
The rapid-deployment valve system (RDVS) was introduced to facility minimally invasive aortic valve replacement. In this study we evaluate the potential benefits of RDVS in elderly high-risk patients with endocarditis of the aortic valve. Since the introduction of RDVS in our institution in December 2017 through October 2021, EDWARDS INTUITY rapid-deployment prosthesis (Model 8300A, Edwards Lifesciences, Irvine, CA, USA) has been implanted in a total of 115 patients for different indications by a single surgeon. Out of one-hundred and fifteen cases of RDVS implantation, seven patients with a median age of 77 yrs. (range 62-84yrs.), suffered from active infective endocarditis of the aortic valve. The median EuroSCORE II of these highly selected patient cohort was 77% (range 19-80%). Patient data were evaluated perioperatively including intra-operative data as well as in-hospital morbidity/mortality and follow-up after discharge from hospital. Three patients underwent previous cardiac surgery. Concomitant procedures were performed in six patients including, ascending aorta replacement (n = 3), mitral valve repair (n = 1), pulmonary valve replacement (n = 1), bypass surgery (n = 1), left atrial appendix resection (n = 1) and anterior mitral valve repair (n = 1). Median aortic cross-clamp and cardiopulmonary bypass time was 56 min (range 29-122 min) and 81 min (range 45-162 min.), respectively. Post-operative complications in these elderly high-risk patients were atrial fibrillation (n = 3) and re-exploration for pericardial effusion (n = 1). One pacemaker implantation was required on postoperative day 6 due to sick sinus syndrome. There was one in-hospital death (14%) and one during follow-up (14%). Rapid-deployment aortic valve system seems to be a viable option with acceptable morbidity and mortality in elderly high-risk patients with active infective endocarditis of the aortic valve.


Immunoglobulin A Antibodies Against Myelin Oligodendrocyte Glycoprotein in a Subgroup of Patients With Central Nervous System Demyelination.
Differential diagnosis of patients with seronegative demyelinating central nervous system (CNS) disease is challenging. In this regard, evidence suggests that immunoglobulin (Ig) A plays a role in the pathogenesis of different autoimmune diseases. Yet little is known about the presence and clinical relevance of IgA antibodies against myelin oligodendrocyte glycoprotein (MOG) in CNS demyelination. To investigate the frequency of MOG-IgA and associated clinical features in patients with demyelinating CNS disease and healthy controls. This longitudinal study comprised 1 discovery and 1 confirmation cohort derived from 5 centers. Participants included patients with suspected or confirmed demyelinating diseases and healthy controls. MOG-IgA, MOG-IgG, and MOG-IgM were measured in serum samples and cerebrospinal fluid (CSF) of patients, who were assessed from September 2012 to April 2022. Frequency and clinical features of patients who were seropositive for MOG-IgA and double-seronegative for aquaporin 4 (AQP4) IgG and MOG-IgG. After the exclusion of 5 participants with coexisting AQP4-IgG and MOG-IgA, MOG-IgG, and/or MOG-IgM, 1339 patients and 110 healthy controls were included; the median follow-up time was 39 months (range, 0-227 months). Of included patients with isolated MOG-IgA, 11 of 18 were female (61%), and the median age was 31.5 years (range, 3-76 years). Among patients double-seronegative for AQP4-IgG and MOG-IgG (1126/1339; 84%), isolated MOG-IgA was identified in 3 of 50 patients (6%) with neuromyelitis optica spectrum disorder, 5 of 228 patients (2%) with other CNS demyelinating diseases, and 10 of 848 patients (1%) with multiple sclerosis but in none of the healthy controls (0/110). The most common disease manifestation in patients seropositive for isolated MOG-IgA was myelitis (11/17 [65%]), followed by more frequent brainstem syndrome (7/16 [44%] vs 14/75 [19%], respectively; P = .048), and infrequent manifestation of optic neuritis (4/15 [27%] vs 46/73 [63%], respectively; P = .02) vs patients with MOG-IgG. Among patients fulfilling 2017 McDonald criteria for multiple sclerosis, MOG-IgA was associated with less frequent CSF-specific oligoclonal bands (4/9 [44%] vs 325/351 [93%], respectively; P < .001) vs patients with multiple sclerosis who were MOG-IgG/IgA seronegative. Further, most patients with isolated MOG-IgA presented clinical attacks after recent infection or vaccination (7/11 [64%]). In this study, MOG-specific IgA was identified in a subgroup of patients who were double-seronegative for AQP4-/MOG-IgG, suggesting that MOG-IgA may be a novel diagnostic biomarker for patients with CNS demyelination.


Neurodevelopmental sequelae associated with gray and white matter changes and their cellular basis: A comparison between Autism Spectrum Disorder, ADHD and dyslexia.
Many psychiatric diseases, such as major depression and schizophrenia, are accompanied by patterns of gray matter and white matter changes in the cortex that may be due to structural pathologies of synapses and their dendrites in the gray matter on the one hand and to pathologies in myelinating oligodendrocytes on the other. Here the possibility has been briefly examined that such a generalization might also hold for Autistic Spectrum Disorders (ASD). Evidence is presented that gray matter changes that accompany ASD may in fact reflect changes in synapses and subsequently of their dendrites, whereas those in the white matter reflect changes in myelination due to pathologies of oligodendrocytes. It is proposed that such structural pathologies during development provide a coherent biological model not only for the onset and course of ASD but also provide the basis for development and systematic evaluation of new treatment strategies.


Simple sequential boundaries for treatment selection in multi-armed randomized clinical trials with a control.
In situations when many regimens are possible candidates for a large phase III study, but too few resources are available to evaluate each relative to the standard, conducting a multi-armed randomized selection trial is a useful strategy to remove inferior treatments from further consideration. When the study has a relatively quick endpoint such as an imaging-based lesion volume change in acute stroke patients, frequent interim monitoring of the trial is ethically and practically appealing to clinicians. In this article, I propose a class of sequential selection boundaries for multi-armed clinical trials, in which the objective is to select a treatment with a clinically significant improvement upon the control group, or to declare futility if no such treatment exists. The proposed boundaries are easy to implement in a blinded fashion, and can be applied on a flexible monitoring schedule in terms of calendar time. Design calibration with respect to prespecified levels of confidence is simple, and can be accomplished when the response rate of the control group is known only up to an interval. One of the proposed methods is applied to redesign a selection trial with an imaging endpoint in acute stroke patients, and is compared to an optimal two-stage design via simulations: The proposed method imposes smaller sample size on average than the two-stage design; this advantage is substantial when there is in fact a superior treatment to the control group.


[Outpatient dermatoplasty in open injuries to the fingers and hand].
A method of skin plasty of tissue defects on hands and fingers by skin flaps on feeding pedicle cut on the femur is described. Under conditions of the out-patient clinic the author has made operations on 12 patients with good results.


The microbiota of farmed mink (Neovison vison) follows a successional development and is affected by early life antibiotic exposure.
On many mink farms, antibiotics are used extensively during the lactation period to reduce the prevalence and severity of pre-weaning diarrhoea (PWD) in mink kits (also referred to as greasy kit syndrome). Concerns have been raised, that routine treatment of PWD with antibiotics could affect the natural successional development of the gut microbiota, which may have long lasting consequences. Here we investigated the effects of early life antibiotic treatment administered for 1 week (postnatal days 13-20). Two routes of antibiotic administration were compared to a non-treated control group (CTR, n = 24). Routes of administration included indirect treatment, through the milk from dams receiving antibiotics by intramuscular administration (ABX_D, n = 24) and direct treatment by intramuscular administration to the kits (ABX_K, n = 24). A tendency for slightly increased weight at termination (Day 205) was observed in the ABX_K group. The gut microbiota composition was profiled by 16S rRNA gene sequencing at eight time points between Day 7 and Day 205. A clear successional development of the gut microbiota composition was observed and both treatment regimens caused detectable changes in the gut microbiota until at least eight days after treatment ceased. At termination, a significant positive correlation was identified between microbial diversity and animal weight.


Drug safety of systemic treatments for psoriasis: results from The German Psoriasis Registry PsoBest.
The German Psoriasis Registry PsoBest was conducted in 2008 in order to investigate the long-term outcomes and safety of systemic treatments for moderate-to-severe psoriasis. Safety analysis of antipsoriatic drugs with special focus on serious adverse events (SAE) for infections, malignancies and major cardiac events (MACE) was done. Nationwide non-interventional patient treatment registry conducted in 251 active dermatology centers. Until June 2012, n = 2444 patients [40 % female; mean age 47.3 (SD 14.1) years; mean duration of disease 18.2 (SD 14.7) years] were recruited, including n = 1791 patients (3842 patient years) with conventional systemic drugs and n = 908 (3442 patient years) with biological drugs. Mean PASI (Psoriasis Area and Severity Index) at inclusion was 14.7, mean DLQI (Dermatology Life Quality Index) 11.1, mean BMI (Body Mass Index) 28.2. The overall rate of SAE per 100 patient years were 1.3 (SD 0.9) per 100 patient years in conventional systemic and 1.5 (SD 1.2) in biologics (p > 0.5, no significant difference). The rates per 100 patient years for single severe adverse events were as follows (systemic/biologics): serious infections, 0.33/0.65 [CI (confidence interval) 0.13-0.54/0.35-0.98]; MACE, 0.56/0.77 (CI 0.29-0.97/0.41-1.31); malignancies (except non-melanoma skin cancer), 0.46/0.49 (CI 0.22-0.84/0.21-0.97). There were no significant differences between single drugs in any of the safety parameters. The conventional systemic and biologic drugs for psoriasis show satisfying safety under routine psoriasis care in Germany with respect to infections, MACE and malignancies.


Translating basic research into clinical rheumatology.
Findings from basic research in combination with precise clinical observations of the disease course in rheumatoid arthritis (RA) have led to the development of a multistage model to explain the pathophysiology of RA. Different cellular and soluble mediators, which play principal roles at different phases of the disease, have been identified. New therapeutic agents, which specifically target these factors, now allow us to intervene at several levels of the pathogenesis. This has already resulted in significant improvements for patients suffering from RA, and the development of new promising agents continues at a high pace. However, many questions concerning the optimal use of the new therapies remain unanswered. Combined efforts of basic research and clinical trials investigating the optimal timing and combination of the new treatments will be necessary to allow them to achieve their full potential and to result in the maximum benefit for patients.


Cell proliferation detected using [18F]FLT PET/CT as an early marker of abdominal aortic aneurysm.
Abdominal aortic aneurysm (AAA) is a focal aortic dilatation progressing towards rupture. Non-invasive AAA-associated cell proliferation biomarkers are not yet established. We investigated the feasibility of the cell proliferation radiotracer, fluorine-18-fluorothymidine ([18F]FLT) with positron emission tomography/computed tomography (PET/CT) in a progressive pre-clinical AAA model (angiotensin II, AngII infusion). Fourteen-week-old apolipoprotein E-knockout (ApoE-/-) mice received saline or AngII via osmotic mini-pumps for 14 (n = 7 and 5, respectively) or 28 (n = 3 and 4, respectively) days and underwent 90-minute dynamic [18F]FLT PET/CT. Organs were harvested from independent cohorts for gamma counting, ultrasound scanning, and western blotting. [18F]FLT uptake was significantly greater in 14- (n = 5) and 28-day (n = 3) AAA than in saline control aortae (n = 5) (P < 0.001), which reduced between days 14 and 28. Whole-organ gamma counting confirmed greater [18F]FLT uptake in 14-day AAA (n = 9) compared to saline-infused aortae (n = 4) (P < 0.05), correlating positively with aortic volume (r = 0.71, P < 0.01). Fourteen-day AAA tissue showed increased expression of thymidine kinase-1, equilibrative nucleoside transporter (ENT)-1, ENT-2, concentrative nucleoside transporter (CNT)-1, and CNT-3 than 28-day AAA and saline control tissues (n = 3 each) (all P < 0.001). [18F]FLT uptake is increased during the active growth phase of the AAA model compared to saline control mice and late-stage AAA.


Effect of oil substitution in chiral microemulsion electrokinetic chromatography.
In a previous publication (Pascoe, R., Foley, J. P., Analyst 2002, 127, 710-714), a novel chiral microemulsion based on 1.0% w/v dodecoxycarbonylvaline (DDCV), 0.50% v/v ethyl acetate and 1.2% v/v 1-butanol, was shown to provide rapid enantiomeric separations of various pharmaceutical compounds. The two deficiencies noted with this method were that the peak shapes obtained were asymmetric and the efficiencies were lower than those previously obtained using DDCV micelles (Peterson, A. G., Ahuja, E. S., Foley, J. P., J. Chromatogr. B 1996, 683, 15-28). This study examines the use of three alternative low-interfacial-tension oils (methyl acetate, methyl propionate, and methyl formate), in combination with DDCV, to characterize their effect on the elution range, efficiency, resolution, and enantioselectivity of various pharmaceutical enantiomers. The oils were evaluated in both the same volume percentage and the same molar concentration as ethyl acetate in the original DDCV microemulsion system. Including ethyl acetate, a total of seven microemulsion systems were examined. For the compounds that were separated, average enantioselectivities ranged from 1.09 to 1.28, with corresponding efficiencies of 14,000-20,000. While some interesting differences were observed, ethyl acetate still proved to be the most advantageous in terms of enantioselectivity, resolution, and elution range.


Function and dysfunction of prefrontal brain circuitry in alcoholic Korsakoff's syndrome.
The signature symptom of alcohol-induced persisting amnestic disorder, more commonly referred to as alcoholic Korsakoff's syndrome (KS), is anterograde amnesia, or memory loss for recent events, and until the mid 20th Century, the putative brain damage was considered to be in diencephalic and medial temporal lobe structures. Overall intelligence, as measured by standardized IQ tests, usually remains intact. Preservation of IQ occurs because memories formed before the onset of prolonged heavy drinking--the types of information and abilities tapped by intelligence tests--remain relatively well preserved compared with memories recently acquired. However, clinical and experimental evidence has shown that neurobehavioral dysfunction in alcoholic patients with KS does include nonmnemonic abilities, and further brain damage involves extensive frontal and limbic circuitries. Among the abnormalities are confabulation, disruption of elements of executive functioning and cognitive control, and emotional impairments. Here, we discuss the relationship between neurobehavioral impairments in KS and alcoholism-related brain damage. More specifically, we examine the role of damage to prefrontal brain systems in the neuropsychological profile of alcoholic KS.


NAD(+) administration decreases ischemic brain damage partially by blocking autophagy in a mouse model of brain ischemia.
Nicotinamide adenine dinuleotide (NAD(+)) plays critical roles in multiple biological functions. Previous studies have indicated that NAD(+) treatment decreases oxidative stress-induced death of primary neurons and astrocytes. Intranasal administration of NAD(+) also reduces brain damage in a rat model of transient focal brain ischemia. However, the mechanisms underlying this protective effect remain unknown. In this study, we used a mouse model of brain ischemia to test our hypothesis that NAD(+) decreases ischemic brain damage partially by preventing autophagy. Adult male mice were subjected to transient middle cerebral artery occlusion (tMCAO) for 90min, and NAD(+) was administered intraperitoneally (i.p.) immediately after reperfusion started. We found that administration with 50mg/kg NAD(+) led to significant decreases in infarct size, edema formation, and neurological deficits at 48h after ischemia. NAD(+) administration also significantly decreased brain ischemia-induced autophagy in the cortex and hippocampus. We further found that prevention of autophagy by 3-methyladenine (3-MA), a selective autophagy inhibitor, significantly reduced ischemic brain damage, suggesting an important role of autophagy in the ischemic brain injury in our animal model. Collectively, our findings have suggested that NAD(+) administration decreases ischemic brain damage at least partially by blocking autophagy. This is the first suggested mechanism regarding the protective effects of NAD(+) in cerebral ischemia, which further highlights the promise of NAD(+) for treating brain ischemia.


Slice-layer COFs-aerogel: a regenerative dispersive solid-phase extraction adsorbent for determination of ultra-trace quinolone antibiotics.
A novel type of three-dimensional network structure, covalent organic frameworks (COFs) aerogel, was fabricated and applied to dispersive solid-phase extraction (dSPE) of quinolone antibiotics (QAs). Density functional theory (DFT) was applied to investigate the possible interaction mechanism and results confirmed that the strong adsorption affinity is attributed to the intralayer hydrogen bonds and π-π interaction. Furthermore, a sensitive analytical method based on COFs-aerogel for determining quinolone antibiotics residues in water and honey samples was developed and HPLC-MS/MS was used for sample detection and quantification. Under the optimal conditions, COFs-aerogel exhibited a wide linearity (0.1-500 ng∙L-1), low limits of detection (0.02-0.06 ng∙L-1), and good precision (RSD ˂ 10%) for selected QAs. A preliminary practical application of the developed method was proved by the efficient detection of quinolone antibiotics in water and food samples with good recoveries (68.2-104% and 64.0-100% for water and honey samples, respectively). Combining the experimental data with theoretical calculation, results illustrated that COFs-aerogel holds a great potential to capture contaminants and address environmental and food safety issues.


Oligodendrocyte lineage is severely affected in human alcohol-exposed foetuses.
Prenatal alcohol exposure is a major cause of neurobehavioral disabilities. MRI studies in humans have shown that alcohol is associated with white matter microstructural anomalies but these studies focused on myelin abnormalities only after birth. Only one of these studies evaluated oligodendrocyte lineage, but only for a short period during human foetal life. As data are lacking in humans and alcohol is known to impair oligodendrocyte differentiation in rodents, the present study aimed to compare by immunohistochemistry the oligodendrocyte precursor cells expressing PDGFR-α and immature premyelinating/mature oligodendrocytes expressing Olig2 in the ganglionic eminences and the frontal cortex of 14 human foetuses exposed to alcohol from 15 to 37 weeks' gestation with age-matched controls. The human brains used in this study were obtained at the time of foetal autopsies for medical termination of pregnancy, in utero or post-natal early death. Before birth, PDGFR-α expression was strongly increased in the ganglionic eminences and the cortex of all foetuses exposed to alcohol except at the earliest stage. No massive generation of Olig2 immunoreactive cells was identified in the ganglionic eminences until the end of pregnancy and the density of Olig2-positive cells within the cortex was consistently lower in foetuses exposed to alcohol than in controls. These antenatal data from humans provides further evidence of major oligodendrocyte lineage impairment at specific and key stages of brain development upon prenatal alcohol exposure including defective or delayed generation and maturation of oligodendrocyte precursors.


Cytochemical demonstration of the molecular forms of cardiac glycosides in the heart muscle.
Selective topo-optical staining of vicinal-OH groups with aldehyde-bisulphite-toluidine blue (ABT) has been used for studying the localization and molecular structural order of cardiac glycoside in heart muscle. The glycoside has strong metachromatic basophilia and negative birefringence. In addition to confirming the electron microscopic suitability of the ABT reaction has offered also ultrastructural evidence for the reliability of the topo-optical method. The localization appeared to be in extracellular connection with the outer side of sarcolemma membrane, internal surface of endothelial cell, and in the subsarcolemmal cysterne. The findings suggest that the topo-optical reaction is suitable for molecular analysis of cardiac glycoside.


Salvador-warts-hippo signaling promotes Drosophila posterior follicle cell maturation downstream of notch.
The Salvador Warts Hippo (SWH) network limits tissue size in Drosophila and vertebrates [1]. Decreased SWH pathway activity gives rise to excess proliferation and reduced apoptosis. The core of the SWH network is composed of two serine/threonine kinases Hippo (Hpo) and Warts (Wts), the scaffold proteins Salvador (Sav) and Mats, and the transcriptional coactivator Yorkie (Yki) [1]. Two band 4.1 related proteins, Merlin (Mer) and Expanded (Ex), have been proposed to act upstream of Hpo, which in turn activates Wts ([1] for review). Wts phosphorylates and inhibits Yki, repressing the expression of Yki target genes [2-4]. Recently, several planar cell polarity (PCP) genes have been implicated in the SWH network in growth control [5-8]. Here, we show that, during oogenesis, the core components of the SWH network are required in posterior follicle cells (PFCs) competent to receive the Gurken (Grk)/TGFalpha signal emitted by the oocyte to control body axis formation. Our results suggest that the SWH network controls the expression of Hindsight, the downstream effector of Notch, required for follicle cell mitotic cycle-endocycle switch. The PCP members of the SWH network are not involved in this process, indicating that signaling upstream of Hpo varies according to developmental context.


MicroRNA-410-3p Binds to TLR2 and Alleviates Myocardial Mitochondrial Dysfunction and Chemokine Production in LPS-Induced Sepsis.
Mitochondrial dysfunction and chemokine production have been reported to be involved in the pathogenesis of sepsis. Our initial bioinformatics analysis identified differentially expressed TLR2 in sepsis and the upstream regulatory microRNA-410-3p (miR-410-3p). Hence, the current study was performed to characterize the potential mechanism by which miR-410-3p modulates mitochondrial dysfunction and chemokine production in lipopolysaccharide (LPS)-induced mice in vivo and cardiomyocytes in vitro. Next, we identified that miR-410-3p was downregulated, while TLR2 was upregulated in LPS-induced mice and cardiomyocytes. In addition, miR-410-3p was confirmed to target and inhibit the TLR2 expression. Thereafter, gain- or loss-of-function experiments were conducted to investigate the effect of miR-410-3p and TLR2 on mitochondrial function and chemokine production. TLR2 knockdown or miR-410-3p overexpression was found to alleviate mitochondrial membrane damage and mitochondrial swelling, in addition to augmenting the levels of adenosine triphosphate, mitochondrial membrane potential, and the expression levels of CCL7, CCL5, CXCL1, and CXCL9 in vivo and in vitro. In conclusion, miR-410-3p-mediated TLR2 inhibition alleviated mitochondrial dysfunction and reduced chemokine production in LPS-induced experimental sepsis. Therefore, the overexpression of miR-410-3p may represent a potential strategy for the treatment of sepsis-induced myocardial injury.


Differential ontogeny of GABA(B)-receptor-mediated pre- and postsynaptic modulation of GABA and glycine transmission in respiratory rhythm-generating network in mouse.
Rhythm generation in mature respiratory networks is influenced strongly by synaptic inhibition. In early neonates, GABA(A)-receptor- and glycine-receptor-mediated inhibition is not present, thus the question arises as to whether GABA(B)-receptor-mediated inhibition plays an important role. Using brainstem slices of neonatal mice (postnatal day, P0-P15), we analysed the role of GABA(B)-mediated modulation of GABA and glycine synaptic transmission in the respiratory network. Blockade of GABA uptake by nipecotic acid (0.25-2 mM) reduced the respiratory frequency. This reduction was prevented by the selective GABA(B) receptor antagonist CGP55845A (CGP) alone at P0-P3, but by bicuculline as well as CGP at P7-P15. Blockade of GABA(B) receptors by CGP increased the respiratory frequency at P0-P3, whereas it caused a reduction of frequency in older animals. The effect of CGP on respiratory frequency was diminished in the presence of bicuculline and strychnine in older but not in younger animals. The relative contribution of GABA(B)-receptor-mediated pre- and postsynaptic modulation was examined by analysing the effect of GABA(B) receptors on spontaneous and miniature IPSCs. In younger animals (P0-P3), the GABA(B) receptor agonist baclofen had no detectable effect on IPSC frequency, but caused a significant decrease in the amplitude. In older animals (P7-P15), baclofen decreased both the frequency and amplitude of spontaneous and miniature IPSCs. These results demonstrate that GABA(B)-receptor-mediated postsynaptic modulation plays an important role in the respiratory network from P0 on. GABA(B)-receptor-mediated presynaptic modulation develops with a longer postnatal latency, and becomes predominant within the first postnatal week.


Multiphoton microscopy is a nondestructive label-free approach to investigate the 3D structure of gas cell walls in bread dough.
During the different steps of bread-making, changes in the microstructure of the dough, particularly in the gas cell walls (GCW), have a major influence on the final bread crumb texture. Investigation of the spatial conformation of GCWs is still a challenge because it requires both high resolutions and 3D depth imaging. The originality of the present work lies in the use of label-free non-destructive multiphoton microscopy (NLOM) to image the 3D structure of GCWs, shedding light on their behavior and organization in wheat bread dough. We demonstrated that second and third harmonic generation (SHG, THG) allow imaging, respectively, of starch granules and interfaces in bread dough, while the gluten matrix was detected via two-photon excitation fluorescence (TPEF). Last, a distinction between the gluten network and starch granules was achieved using gluten endogenous fluorescence (EF) imaging, while the position, size, and 3D orientation of starch granules in GCWs were determined from harmonic imaging, made possible by the acquisition of backward and forward SHG with linear polarization. These innovative experiments highlight the strengths of NLOM for a label-free characterization of bread dough microstructure for the first time, in order to understand the role of starch granules in dough stabilization.


Effects of high-heeled footwear on static and dynamic pelvis position and lumbar lordosis in experienced younger and middle-aged women.
There is still conflicting evidence about the effect of high-heeled footwear on posture, especially if methodological confounders are taken into account. The purpose of this study was to investigate the effect of high-heeled footwear on lumbopelvic parameters in experienced younger and middle-aged women while standing and walking. Thirty-seven experienced younger (n=19:18-25 years) and middle-aged (n=18:26-56 years) women were included in this randomized crossover study. Using a non-invasive back shape reconstruction device (rasterstereography), static (pelvic tilt and lumbar lordosis angle) and dynamic (pelvic rotation, median lumbar lordosis angle and range of motion) parameters representing pelvis position and lumbar curvature were measured. In order to analyse standing and walking on a treadmill (0.83m/s), the effects of high-heels (7-11cm) were compared to standard control shoes. There were no effects on the lumbar lordosis angle or range of motion under static or dynamic conditions (p>0.05, d≤0.06). But there was a small effect for a reduced pelvic tilt (p=0.003, d=0.24) and a moderate effect for an increased transversal pelvic rotation (p=0.001, d=0.63) due to high heel shoed standing or walking, respectively. There were no significant age-group or interaction effects (p>0.05). Altered pelvic parameters may be interpreted as compensatory adaptations to high-heeled footwear rather than lumbar lordosis adaptations in experienced wearers. The impact of these findings on back complaints should be revisited carefully, because muscular overuse as well as postural load relieving may contribute to chronic consequences. Further research is necessary to examine clinically relevant outcomes corresponding to postural alterations.


Synthesis and biological evaluations of novel benzimidazoles as potential antibacterial agents.
A series of novel benzimidazole derivatives were synthesized via parallel solution-phase chemistry. Many of these compounds were found to inhibit the growth of Staphylococcus aureus and Escherichia coli. Several analogues exhibited low micromolar minimal inhibitory concentrations (MIC) against both Gram-positive and Gram-negative bacteria of clinical relevance and could serve as leads for further optimizations for antibacterial research.


Systematic analysis of occurrence and variation tendency about 58 typical veterinary antibiotics during animal wastewater disposal processes in Tianjin, China.
Residue of veterinary antibiotics (VAs) in the animal breeding industry has become a problematic environmental issue. However, the residual levels of VAs as well as their variation tendency, degradation mechanisms and relationships with other parameters during animal wastewater disposal processes are still obscure. This study measured different samples during wastewater disposal processes from three farms, and systematically analyzed the residue, migration and removal of 58 kinds of typical VAs (6 classes) in Tianjin, China. The results showed that about 44 kinds of VAs were quantitatively detected. Tetracycline antibiotics (TCs) usually had higher residual concentrations than other classes of VAs in the raw wastewater; the highest residual concentration was 130.67 ± 5.90 μg/L which occurred for chlortetracycline (CTC). Pig farms generally had more VAs species and higher residual concentrations than dairy farms, and the proportion of different VAs was similar for dairy farms. The final removal rates of different VAs classes varied largely (negative to > 99.87%), and the highest removal rates usually occurred in biological processes for adsorption and biodegradation effects, and occasionally occurred in the final effluents. The correlation coefficients between VAs removal rates and chemical oxygen demand (COD) removal rates were much higher than those of total nitrogen (TN), total phosphorus (TP) and ammonia nitrogen (NH4-N) in pig farms, while opposite conclusion was obtained in dairy farms. Among different classes, TCs presented the highest daily mass loading of ND ~ 10,453.8 ± 471.7 mg/d in the influent and ND ~ 1141.6 ± 58.9 mg/d in the effluent in farm 1.


Pyelo-choledochal fistula accompanying operative cholangiography.
Fistulous communications between the biliary system and the urinary tract are encountered infrequently and, routinely, they are associated with operative dissections involving both systems. We report a case of opacification of the right renal collecting system during cholangiography following uneventful cholecystectomy. Prompt spontaneous resolution was verified by subsequent excretory urography.


The Role of Biomarkers in Acute Kidney Injury.
Several biomarkers have been developed to detect acute kidney injury (AKI) and predict outcomes. Most AKI biomarkers have been shown to be expressed before serum creatinine and to be more sensitive and specific than urine output. Only a few studies have examined how implementation can change clinical outcomes. A second generation of AKI biomarkers have been developed. These markers, including tissue inhibitor of metalloproteinases-2 (TIMP-2) and insulinlike growth factor-binding protein 7 (IGFBP7), have obtained regulatory approval in many countries based on large, rigorous clinical studies and small, single-centered trials and have begun to establish clinical utility.


Caring for Older Adults With Vision Impairment and Dementia.
Dementia and vision impairment (VI) are common among older adults but little is known about caregiving in this context. We used data from the 2011 National Health and Aging Trends Study, a nationally representative survey of Medicare beneficiaries, linked to their family/unpaid helpers from the National Study of Caregiving. Vision impairment was defined as self-reported blindness or difficulty with distance/near vision. Probable dementia was based on survey report, interviews, and cognitive tests. Our outcomes included hours of care provided, and number of valued activities (scored 0-4) affected by caregiving, per month. Among 1,776 caregivers, 898 (55.1%, weighted) assisted older adults without dementia or VI, 450 (21.9%) with dementia only, 224 (13.0%) with VI only, and 204 (10.0%) with dementia and VI. In fully adjusted negative binomial regression analyses, caregivers of individuals with dementia and VI spent 1.7 times as many hours (95% confidence interval [CI] = 1.4-2.2) providing care than caregivers of those without either impairment; however, caregivers of individuals with dementia only (95% CI = 1.1-1.6) and VI only (95% CI = 1.1-1.6) spent 1.3 times more hours. Additionally, caregivers of individuals with dementia and VI had 3.2 times as many valued activities affected (95% CI = 2.2-4.6), while caregivers of dementia only and VI only reported 1.9 times (95% CI = 1.4-2.6) and 1.3 times (95% CI = 0.9-1.8) more activities affected, respectively. Our results suggest that caring for older adults with VI involves similar time demands as caring for older adults with dementia, but that participation impacts are greater when caring for older adults with both dementia and VI.


A new "closed" in situ vein bypass technique results in a reduced wound complication rate.
This prospective randomised multicentre trial was conducted to test whether a new "closed" technique for in situ vein bypass would result in a lower frequency of wound complications, without negative effects on patency rates and without an intolerable increase in residual arteriovenous fistulae compared to the conventional "open" technique. We have developed a new "closed" technique using a co-axial catheter embolisation system for intra-operative coil embolisation of side branches, in order to avoid long incisions. In four centres and 95 patients, 97 in situ bypasses were performed: 47 "closed" and 50 "open". Randomisation was stratified for below knee femoropopliteal bypasses (60) and femorocrural bypasses (37). Indications were disabling intermittent claudication (29), restpain (26) or ulcers and/or necrosis (42). Postoperative mortality was 2% (one in the "closed", one in the "open" group). A total number of 16 (34%) wound complications (grade 1, 2 and 3) occurred in the closed group compared to 36 (72%) in the open group (p < 0.05). Deep wound complications (grade 2) occurred in six patients (13%) of the "closed" group, compared to 15 (30%) in the "open" group. In both groups, three patients (6%) developed deep wound complications including the bypass area (grade 3). In the "closed" group, 20 patients needed additional treatment for arteriovenous fistulae, compared to four in the "open" group. One-year patency rates did not show a statistically significant difference: primary patency rates were 65% and 61% and secondary patency rates were 86% and 76% respectively for the "closed" and "open" group. These results indicate that a "closed" technique reduces wound complication rate, without negative effects on the short term patency rates. The "closed" technique results in an increased number of postoperative treatments for residual arteriovenous fistulae.


A hybrid approach for patent ductus arteriosus closure using the Amplatzer Duct Occluder.
We performed closure of the patent ductus arteriosus (PDA) using a hybrid approach with an Amplatzer Duct Occluder. Six patients (two males and four females) underwent PDA closure at a mean age of 7.8 months (range 2-24 months) and a mean weight of 6.6 kg (range 4.5-13 kg). The main pulmonary artery (MPA) was exposed via a minimally invasive left parasternal second intercostal space incision. Under transesophageal echocardiography guidance, the PDA occluder was implanted via direct puncture of the MPA. The procedure was successful in all patients with no residual shunt. There were no hospital deaths, and the postoperative course was uneventful. All patients were discharged on the 3rd to 4th day. There was no residual shunt in any patient on midterm follow-up. The novel hybrid approach is a safe, minimal invasive procedure. Further experience and longer follow-up of these patients is necessary to conclude whether this technique is applicable to all the patients with a PDA.


DNA asymmetry in stem cells - immortal or mortal?
The immortal strand hypothesis proposes that stem cells retain a template copy of genomic DNA (i.e. an 'immortal strand') to avoid replication-induced mutations. An alternative hypothesis suggests that certain cells segregate sister chromatids non-randomly to transmit distinct epigenetic information. However, this area of research has been highly controversial, with conflicting data even from the same cell types. Moreover, historically, the same term of 'non-random sister chromatid segregation' or 'biased sister chromatid segregation' has been used to indicate distinct biological processes, generating a confusion in the biological significance and potential mechanism of each phenomenon. Here, we discuss the models of non-random sister chromatid segregation, and we explore the strengths and limitations of the various techniques and experimental model systems used to study this question. We also describe our recent study on Drosophila male germline stem cells, where sister chromatids of X and Y chromosomes are segregated non-randomly during cell division. We aim to integrate the existing evidence to speculate on the underlying mechanisms and biological relevance of this long-standing observation on non-random sister chromatid segregation.


Insulin resistance, cognition and Alzheimer's disease biomarkers: Evidence that CSF Aβ42 moderates the association between insulin resistance and increased CSF tau levels.
Mounting evidence implicates insulin resistance (IR) with reduced cognition, increased dementia risk and changes in Alzheimer's disease biomarkers. It's unclear how, and at what stage IR has the greatest impact on Alzheimer's disease biomarker progression indicative of cognitive decline. Exploration of potential factors influencing this relationship continue. We have previously reported IR to be associated with cognitive function, and increased CSF tau in a cognitively unimpaired cohort. Now, we aimed to determine if CSF total (t-tau) or phosphorylated tau (p-tau) mediated the relationship between HOMA-IR and cognition, and explore sex or amyloid-β (Aβ) biomarkers as moderators of this relationship. Mediation analysis demonstrated that CSF tau does not directly influence the association between HOMA-IR and cognition. Moderation analysis revealed CSF Aβ42 moderates the relationships between HOMA-IR and CSF tau. The combination of lower CSF Aβ42 and higher HOMA-IR was associated with increases in CSF tau. The CSF Aβ42 moderation finding has potential to be considered when assessing type 2 diabetic risk for tau pathology and cognitive decline.


ALK inhibitor resistance in ALK(F1174L)-driven neuroblastoma is associated with AXL activation and induction of EMT.
The crizotinib-resistant ALK(F1174L) mutation arises de novo in neuroblastoma (NB) and is acquired in ALK translocation-driven cancers, lending impetus to the development of novel anaplastic lymphoma kinase (ALK) inhibitors with different modes of action. The diaminopyrimidine TAE684 and its derivative ceritinib (LDK378), which are structurally distinct from crizotinib, are active against NB cells expressing ALK(F1174L). Here we demonstrate acquired resistance to TAE684 and LDK378 in ALK(F1174L)-driven human NB cells that is linked to overexpression and activation of the AXL tyrosine kinase and epithelial-to-mesenchymal transition (EMT). AXL phosphorylation conferred TAE684 resistance to NB cells through upregulated extracellular signal-regulated kinase (ERK) signaling. Inhibition of AXL partly rescued TAE684 resistance, resensitizing these cells to this compound. AXL activation in resistant cells was mediated through increased expression of the active form of its ligand, GAS6, that also served to stabilize the AXL protein. Although ectopic expression of AXL and TWIST2 individually in TAE684-sensitive parental cells led to the elevated expression of mesenchymal markers and invasive capacity, only AXL overexpression induced resistance to TAE684 as well. TAE684-resistant cells showed greater sensitivity to HSP90 inhibition than did their parental counterparts, with downregulation of AXL and AXL-mediated ERK signaling. Our studies indicate that aberrant AXL signaling and development of an EMT phenotype underlie resistance of ALK(F1174L)-driven NB cells to TAE684 and its derivatives. We suggest that the combination of ALK and AXL or HSP90 inhibitors be considered to delay the emergence of such resistance.


Microdeletion 8q22.2-q22.3 in a 40-year-old male.
Interstitial deletions at chromosome 8q22.2-q22.3 have been rarely reported in the literature. To date, six patients have been described in the literature with deletions varying in size from 1.36 Mb to 6.44 Mb. These patients range in age from early childhood to early adulthood. The interstitial deletion phenotype has been described to involve moderate to severe intellectual disability, seizures and a distinct facial phenotype. We report on a 40-year-old male with a 3.351 Mb deletion at chromosome 8q22.2-q22.3 who presents with moderate intellectual disability, autism spectrum disorder, childhood seizure disorder, congenital heart defect and hearing loss. He is the oldest known patient to date. Array comparative genomic hybridization (aCGH) was performed on DNA extracted from peripheral blood. This is the first report of an individual with chromosome 8q22.2-q22.3 interstitial deletion associated with congenital heart disease and hearing loss. Haploinsufficiency of the GRHL2 gene contained within the microdeletion is proposed as a candidate genetic mechanism for this patient's hearing loss.


Urothelial bladder afferents selectively project to L6/S1 levels and are more peptidergic than those projecting to the T13/L1 levels in female rats.
This neuroanatomical study in four, adult, Sprague-Dawley female rats quantified the number of Urothelial (labeled by intravesical DiI dye administration) and Non-Urothelial (labeled by intraparenchymal injection of Fast blue dye) bladder primary afferent neurons (bPANs) located in the T13, L1, L6 and S1 dorsal root ganglia. Additional immunohistochemical labeling using antibodies to detect either Substance P or CGRP further characterized the bPAN samples as peptidergic or non-peptidergic. Cell counts indicated that Urothelial bPANs were more common at the L6/S1 levels and more likely to be identified as peptidergic when compared with bPANs characterized at T13/L1 levels and with Non-Urothelial bPANs. These studies provide additional evidence that at least two distinct neuronal populations, with differing localization of sensory terminals, differing peptide content, and differing projections to the central nervous system, are responsible for bladder sensation.


Blood and aqueous humor phoenixin, endocan and spexin in patients with diabetes mellitus and cataract with and without diabetic retinopathy.
Peptide phoenixin (PNX), endocan (EDC), and spexin (SPX) are associated with diabetes. Therefore, the purpose of this study was to investigate the levels of PNX, EDC and SPX in the blood and aqueous humor (AH) of patient with type 2 diabetes with and without DRP and cataract. 30 type 2 diabetes patients with cataract (DM + C), 30 DRP patient with cataract (DRP + C), 30 non-diabetic patient with only cataract and 30 control participants were enrolled into this study. PNX, EDC, and SPX were measured in blood and AH by ELISA. In patients with DRP + C, the levels of PNX and EDC were significantly higher in both AH and blood samples compared with the group of patients without DRP + C (<0.05). Also, in patients with DM + C, the levels of PNX and EDC were higher in both AH and blood samples compared with the group of patients without DM + C. However, in patients with DRP + C, the levels of SPX were significantly lower in both AH and blood samples compared with the group of patients without DRP + C (<0.05). Also, in patients with DM + C, the levels of SPX were also lower in both AH and blood samples compared with the group of patients without DM + C. These findings suggest that increased PNX, EDC, and decreased SPX amounts in blood and AH of DM + C and DRP + C groups when compared with control and cataract groups show that they might have a role in the pathophysiology of DM + C, especially in the DRP + C.


Implementation and validation of real-time algorithms for atrial fibrillation detection on a wearable ECG device.
Due to the growing epidemic of atrial fibrillation (AF), new strategies for AF screening, diagnosis, and monitoring are required. Wearable devices with on-board AF detection algorithms may improve early diagnosis and therapy outcomes. In this work, we implemented optimized algorithms for AF detection on a wearable ECG monitoring device and assessed their performance. The signal processing framework was composed of two main modules: 1) a QRS detector based on a finite state machine, and 2) an AF detector based on the Shannon entropy of the symbolic word series obtained from the instantaneous heart rate. The AF detector was optimized off-line by tuning its parameters to reduce the computational burden while preserving detection accuracy. On-board performance was assessed in terms of detection accuracy, memory usage, and computation time. The on-board implementation of the QRS detector produced an overall accuracy of 99% on the MIT-BIH Arrhythmia Database, with memory usage = 672 bytes, and computation time ≤90 μs. The on-board implementation of the optimized AF algorithm gave an overall accuracy of 98.1% (versus 98.3% of the original version) on the MIT-BIH AF Database, with increased sensitivity (99.2% versus 98.5%) and decreased specificity (97.3% versus 98.2%), memory usage = 4648 bytes, and computation time ≤ 75 μs (consistent with real-time detection). This study demonstrated the feasibility of real-time AF detection on a wearable ECG device. It constitutes a promising step towards the development of novel ECG monitoring systems to tackle the growing AF epidemic.


A study on the membrane depolarization of skeletal muscles caused by a scorpion toxin, sea anemone toxin II and crotamine and the interaction between toxins.
Quinquestriatus toxin (QTX) isolated from the venom of a scorpion (Leiurus quinquestriatus) and sea anemone (Anemonia sulcata) toxin II enhanced the twitch response of the rat and mouse diaphragms and like crotamine (isolated from the venom of Crotalus durissus terrificus) caused spontaneous fasciculation of the muscle. Trains of action potentials in muscles at 70-250 Hz, which could not be antagonized by (+)-tubocurarine, were triggered by single stimulation or occurred spontaneously after treatment with these toxins. QTX and toxin II prolonged the rat muscle action potential 3 to 4 fold whereas crotamine prolonged the action potential by only 30%. The membrane potential was depolarized from about -82 mV to -55 mV by crotamine 2 micrograms ml-1, -41 mV by toxin II 5 micrograms ml-1 and to -50 mV by QTX 1 microgram ml-1. The concentrations to induce 50% maximal depolarization (K0.5) were 0.07, 0.15 and greater than 0.4 microgram ml-1, respectively, for QTX, crotamine and toxin II, whereas the rates of depolarization were in the order toxin II greater than or equal to crotamine greater than QTX. The depolarizing effects of crotamine and QTX, but not of toxin II, were saturable. The depolarizing effects of all three toxins were irreversible whereas the membrane potential could be restored by tetrodotoxin non-competitively. Simultaneous treatment with crotamine and QTX or crotamine and toxin II at concentrations below K0.5 caused only additive effects on depolarization. When the muscle was depolarized by pretreating with a saturating concentration of crotamine, the onset of depolarization by QTX was greatly retarded whereas that by toxin II was unaffected. Action potentials were further prolonged in both cases. 8 It is inferred that all three peptide toxins act at sites on the sodium channel and the binding sites for QTX and crotamine overlap to a considerable extent. On the other hand, the site for toxin II appears not to overlap with that of crotamine but may overlap with that of QTX.


Comparative study of commercial media to improve GMP manufacturing of recombinant human interferon β-1a by CHO cells in perfusion bioreactor.
Chinese hamster ovary cells are the main cellular factories for production of a wide range of recombinant proteins in biopharmaceutical industry. Recombinant human Interferon beta-1a (rh-IFN β-1a), as a cytokine is broadly used to treat multiple sclerosis. In this work, the cell line producing rh-IFN β-1a was studied to improve cell density along with the specific expression. For this reason different cell culture experiments were done using different commercial serum-free media to find the appropriate media providing higher cell density. It was shown DMEMF12, DMEM:ProCHO5, and CHO-S-SFM II led to higher cell density and shorter doubling time. Next, using these media, fed-batch, and perfusion culture with temperature shift were implemented to investigate the best condition for industrial-scale manufacturing of rh-IFN β-1a in terms of higher cell density and product expression yield. The results demonstrated that CHO-S-SFM II media and a thermally biphasic condition provide enhanced expression of rh-IFN β-1a in perfusion bioreactor.


Exploring the impact of cattle on human exposure to malaria mosquitoes in the Arba Minch area district of southwest Ethiopia.
The success of indoor interventions that target mosquitoes for malaria control is partially dependent on early evening and outdoor biting behaviours of mosquito vectors. In southwest Ethiopia, people and cattle live in proximity, which calls to investigate whether the presence of cattle increase or decrease bites from malaria mosquito vectors. This study assessed both host-seeking and overnight activity of malaria mosquito vectors given the presence or absence of cattle in Chano Mille village, Arba Minch district, Ethiopia. Anopheles species density and activity time was compared when a calf was: (i) placed inside; (ii) 1 m away from; or (iii) absent from a tent with a human volunteer resting insides using hourly human landing catches (HLC) conducted from 18:00-0:00 h for 3 months. This trial was performed close to the shore of the Lake Abaya to minimize the interference of other animals on mosquito movement. The overnight activity of malaria vectors was assessed within a Chano village from 18:00-6:00 h with collections carried out both indoors and outdoors by HLC. Generalized estimating equations were used to statistically assess differences. Anopheles pharoensis was significantly more prevalent when a calf was present either inside (42%, P < 0.001), or adjacent to (46%, P = 0.002) a tent relative to a tent without a calf present. The presence of a calf did not affect densities of the primarily anthropophilic species A. gambiae (s.l.), or An. tenebrosus. Anopheles gambiae (s.l.) (P < 0.001) and An. pharoensis (P = 0.015) both had a tendency for early evening biting between 19:00 h and 20:00 h. Anopheles gambiae (s.l.) was mainly biting humans outdoors in the village. The presence of calves within and close to human dwellings acts to draw malaria mosquitoes toward the human occupant with the potential to increase their risk of malaria. Hence, deployment of cattle far from human residence could be recommended to reduce human exposure. Outdoor and early evening biting could threaten the success of current indoor-based interventions. Hence, tools could be designed to reduce this threat.


Cognitive, language and motor development of infants exposed to risk and protective factors.
Risk factors are biological or environmental characteristics increasing the likelihood of delays in child development. Meanwhile, protective factors are conditions that can minimize risks and favor the acquisition of skills. Infants with risk indicators for hearing loss (RIHL) tend to live in less stimulating environments which may lead to lower cognitive, language, and motor development. To compare the cognitive, language, and motor development of infants under the influence of risk and protective factors. A cross-sectional observational study in which 259 infants aged 8-10 months were assessed for cognitive, language, and motor development using the Bayley Scales of Infant and Toddler Development - Third Edition (BSITD-III). The groups were formed according to the presence or absence of RIHL and the quality of the resources, being: SG-AE (Study Group with Adequate Environment), SG-IE (Study Group with Inadequate Environment), CG-AE (Compared Group with Adequate Environment)) and CG-IE (Compared Group with Inadequate Environment). Affordances were assessed using the questionnaire Affordances in the Home Environment for the Motor Development - Infant Scale (AHEMD-IS). The groups were compared using the Chi-square test or the Kruskal-Wallis test, followed by Dunn's multiple comparison test. The level of significance adopted was 5%. Infants from the CG-AE performed better than the other groups in cognition and motor skills. The CG-AE language was statistically superior to the SG-IE. SG-IE showed the highest frequency of delays in all domains. CG-IE and SG-AE showed a similar frequency of delays. Adequate environment associated with the absence of RIHL (CG-AE) leads to better cognitive, language, and motor performances. Biological and environmental risk factors have a similar impact on development, but the accumulation of both tends to increase the risks of developmental delay. The absence of RIHL and quality environments worked as protective factors and favored the acquisition of skills.


Qualitative research in dentistry.
This paper is the first in a series of four that provides an overview of the key elements of qualitative research. In particular, it discusses issues such as what qualitative research is, when its use is appropriate, what it can offer dentistry and approaches to data collection and analysis. Where appropriate, examples of dental studies that have used qualitative methods are also provided for practical purposes.


Effects of Raman scattering across the visible spectrum in clear ocean water: a Monte Carlo study.
Raman-scattering activity in clear ocean waters is documented for the visible spectrum from Monte Carlo simulations. The Raman-scattering activity has a significant effect on the upwelling irradiance value in air and on the submarine light field at the water surface across the visible spectrum. A reduction in Raman-scattering activity at 440 nm that is due to Fraunhofer lines at the Raman source wavelengths is also demonstrated. At wavelengths greater than 500 nm, Raman scattering makes a significant contribution to the in-water light field at depth.


An association study of 13 SNPs from seven candidate genes with pediatric asthma and a preliminary study for genetic testing by multiple variants in Taiwanese population.
Asthma is one of the most common chronic diseases in children. It is caused by complex interactions between various genetic factors and exposures to environmental allergens and irritants. Because of the heterogeneity of the disease and the genetic and cultural differences among different populations, a proper association study and genetic testing for asthma and susceptibility genes is difficult to perform. We assessed 13 single-nucleotide polymorphisms (SNPs) in seven well-known asthma susceptibility genes and looked for association with pediatric asthma using 449 asthmatic subjects and 512 non-asthma subjects in Taiwanese population. CD14-159 C/T and MS4A2 Glu237Gly were identified to have difference in genotype/allele frequencies between the control group and asthma patients. Moreover, the genotype synergistic analysis showed that the co-contribution of two functional SNPs was riskier or more protective from asthma attack. Our study provided a genotype synergistic method for studying gene-gene interaction on polymorphism basis and genetic testing using multiple polymorphisms.


Premenopausal women with breast cancer in the early post-partum period show molecular profiles of invasion and are associated with poor prognosis.
Young premenopausal women develop breast cancer (BC) within 5-10 years of the last childbirth, known as post-partum breast cancers (PPBC), often present with aggressive disease. The exact mechanisms that lead to poor prognosis in these patients are largely unknown. We have evaluated the association of clinical and reproductive factors with BC in a cohort of women ≤ 45 years (N = 155) with long-term follow-up. Based on duration since last childbirth (LCB), grouped patients into PPBC1 (LCB ≤ 5 years), PPBC2 (LCB between 6 and 10 years), PPBC3 (LCB > 10 years), and NPBC (age-matched nulliparous BC patients). We compared disease-free survival and hazard associated with recurrence/metastasis between the groups. RNA sequencing of tumor samples was performed from three parous groups (n = 10), and transcriptomic data were analyzed for differentially expressed genes and altered pathways. Women in the PPBC1 group had an early menarche and late age at first and last childbirth compared to other groups. Survival analysis within lymph node-positive tumors showed that PPBC1 tumors had a worse prognosis than PPBC2 and NPBC tumors (p = 0.015 and p = 0.026, respectively). Clustering of the differentially expressed genes between the groups showed distinct expression in early PPBC (E-PPBC) tumors. Pathway analysis revealed upregulation of invasive-related pathways along with T cell exhaustion, extracellular matrix remodeling, angiogenesis, and epithelial-to-mesenchymal transition in E-PPBC tumors. Early PPBC is a unique subtype with aggressive clinical features and distinct biology. Further research is needed to accurately project the risk of recurrence and optimal treatment strategies in these young patients.


Pyridinium 1,4-zwitterionic thiolates as a useful class of sulfur-containing synthons: application to the synthesis of 2,5-dihydro-1,4,5-thiadiazepines.
A novel [4+3] cascade cyclization reaction of pyridinium 1,4-zwitterionic thiolates with azoalkenes derived from α-halo hydrazones in situ has been developed. This reaction was found to allow expedient access to an array of 2,5-dihydro-1,4,5-thiadiazepines. Libraries of highly functionalized sulfoxide and sulfone analogues of 2,5-dihydro-1,4,5-thiadiazepines were also obtained via selective oxidation with m-CPBA.


Molecular Detection of Human Adenovirus and Rotavirus in Feces of White-Eared Opossums.
The white-eared opossums (Didelphis albiventris) is the largest Brazilian marsupial and a great example of animal synanthropy. Considering the high potential as a carrier of viruses originating from environmental contamination, the presence of Human adenovirus (AdV) and rotavirus was investigated in the feces of rescued white-eared opossums, which were in the process of rehabilitation. The feces of 49 animals were initially investigated by immunochromatography, with three samples positive for AdV and one sample positive for rotavirus. When submitted to PCR and nested PCR, the samples of six animals were positive for AdV and three animals were positive for group A rotavirus. Two positive samples in the immunochromatographic assay were not confirmed by PCR. After sequencing and phylogenetic analysis of AdV samples, all were identified within the genus Mastadenovirus, one being HAdV-C, four HAdV-E, and one being similar to a Mastadenovirus found in primates. This is the first report of molecular confirmation of human adenovirus and rotavirus in white-eared opossums. These data could be important of anticipation some emerging diseases and their effects on ecosystems health.


Use of stellate ganglion block to salvage an ischemic hand caused by the extravasation of vasopressors.
This case report describes the successful use of early stellate ganglion block to salvage an acutely ischemic hand caused by the extravasation of vasopressors. A young man with a gunshot wound to the right inguinal area was brought to the operating room for surgical hemostasis and exploration. After discovery that the central line had been inserted in the proximity of the area of injury (right femoral vein), the vasopressor infusions were changed to a 14-gauge intravenous line inserted in the dorsum of the right hand. When the intravenous line became infiltrated, the hand was found to be swollen, cold, and underperfused. A right stellate ganglion block was immediately performed to overcome the intense peripheral vasoconstriction and, thus, re-establish circulation to the hand. Stellate ganglion block may prove to be an early measure in the treatment of upper-extremity ischemia caused by vasopressor extravasation.


Dynamically regulated two-site interaction of viral RNA to capture host translation initiation factor.
Many RNA viruses employ internal ribosome entry sites (IRESs) in their genomic RNA to commandeer the host's translational machinery for replication. The IRES from encephalomyocarditis virus (EMCV) interacts with eukaryotic translation initiation factor 4 G (eIF4G), recruiting the ribosomal subunit for translation. Here, we analyze the three-dimensional structure of the complex composed of EMCV IRES, the HEAT1 domain fragment of eIF4G, and eIF4A, by cryo-electron microscopy. Two distinct eIF4G-interacting domains on the IRES are identified, and complex formation changes the angle therebetween. Further, we explore the dynamics of these domains by using solution NMR spectroscopy, revealing conformational equilibria in the microsecond to millisecond timescale. In the lowly-populated conformations, the base-pairing register of one domain is shifted with the structural transition of the three-way junction, as in the complex structure. Our study provides insights into the viral RNA's sophisticated strategy for optimal docking to hijack the host protein.


Beneficiary reported experience and voluntary disenrollment in Medicare managed care.
Disenrollment rates have often been used as indicators of health plan quality, because they are readily available and easily understood by purchasers, health plans, and consumers. Over the past few years, however, indicators that more directly measure technical quality and consumer experiences with care have become available. In this observational study, we examined the relationship between voluntary disenrollment rates from Medicare managed care (MMC) plans and other measures of health plan quality. The results demonstrate that voluntary disenrollment rates are strongly related to direct measures of patient experiences with care and are an important complement to other measures of health plan performance.


[Immunoprophylaxis and serodiagnostic studies of abortion in cows caused by bacteria of the Mimeae group and the genus Yersinia].
Tested was the prophylactic effect of a vaccine against abortions in cows caused by bacteria immunologically related to R Brucella organisms. Results pointed to the substantial drop in the number of abortions in the vaccinated animals. Reported are also results of serologic investigations by the complement-fixation test with cows showing abortions and sterility.


Tissue optimization strategies for high-quality ex vivo diffusion imaging.
Ex vivo diffusion imaging can be used to study healthy and pathological tissue microstructure in the rodent brain with high resolution, providing a link between in vivo MRI and ex vivo microscopy techniques. Major challenges for the successful acquisition of ex vivo diffusion imaging data however are changes in the relaxivity and diffusivity of brain tissue following perfusion fixation. In this study we address this question by examining the combined effects of tissue preparation factors that influence signal-to-noise ratio (SNR) and consequently image quality, including fixative concentration, contrast agent concentration and tissue rehydration time. We present an optimization strategy combining these factors to manipulate the T 1 and T 2 of fixed tissue and maximize SNR efficiency. We apply this strategy in the rat brain, for a diffusion-weighted spin echo protocol with TE = 27 ms on a 9.4 T scanner with a 39 mm volume coil and 660 mT/m 114 mm gradient insert. We used a reduced fixative concentration of 2% paraformaldehyde (PFA), rehydration time more than 20 days, 15 mM Gd-DTPA in perfusate and TR 250 ms. This resulted in a doubling of SNR and an increase in SNR per unit time of 135% in cortical grey matter and 88% in white matter compared with 4% PFA and no contrast agent. This improved SNR efficiency enabled the acquisition of excellent-quality high-resolution (78 μ m isotropic voxel size) diffusion data with b = 4000 s/mm 2 , 30 diffusion directions and a field of view of 40 × 13 × 18 mm3 in less than 4 days. It was also possible to achieve comparable data quality for a standard resolution (150 μ m) diffusion dataset in 2 1 4 h. In conclusion, the tissue optimization strategy presented here may be used to improve SNR, increase spatial resolution and/or allow faster acquisitions in preclinical ex vivo diffusion MRI experiments.


Effects of di-(2-ethylhexyl) phthalate and four of its metabolites on steroidogenesis in MA-10 cells.
Phthalate plasticizers are used in the plastics industry to aid in processing and impart flexibility to plastics. Due to the broad use of plastics, and the tendency of plasticizers to leach out of polymers, plasticizers have become ubiquitous in the environment. Concerns about the testicular toxicity of phthalate plasticizers, in particular di-(2-ethylhexyl) phthalate (DEHP), have arisen due to their ability to cause male reproductive tract abnormalities in animal models. It has been assumed that the DEHP metabolite, mono-(2-ethylhexyl) phthalate (MEHP), is the active compound, however, metabolites such as 2-ethylhexanol, 2-ethylhexanal and 2-ethylhexanoic acid, have not been thoroughly investigated. The aim of this study was to evaluate the anti-androgenic potential of these metabolites in vitro with a mouse Leydig tumor cell line, MA-10 cells. DEHP, MEHP and 2-ethylhexanal were found to decrease cell viability, as well as steroidogenic potential. The latter was assessed using an enzyme-linked immunosorbent assay (ELISA) to quantify steroid production and quantitative real-time polymerase chain reaction (qRT-PCR) to assess gene expression analysis of key steroidogenic enzymes. 2-Ethylhexanal proved to be the most potent steroidogenic disruptor, offering intriguing implications in the search for the mechanism of phthalate testicular toxicity. Overall, the study suggests the involvement of multiple active metabolites in the testicular toxicity of DEHP.


Zearalenone contamination of the aquatic environment as a result of its presence in crops.
The aim of this study was to establish a relation between zearalenone contamination of crops in the Polish province of Wielkopolska and its occurrence in aquatic ecosystems close by the crop fields. Water samples were collected from water bodies such as drainage ditches, wells, or watercourses located in four agricultural areas. Moreover, control water samples were collected from the Bogdanka river, which was located outside the agricultural areas and near an urban area. Cereal samples were collected in the harvest season from each agricultural area close to tested water bodies. Zearalenone (ZEA) was found in all water and cereal samples. The highest concentrations were recorded in the postharvest season (September to October) and the lowest in the winter and spring. Mean ZEA concentrations in water ranged between 1.0 ng L(-1) and 80.6 ng L(-1), and in cereals from 3.72 ng g(-1) to 28.97 ng g(-1). Our results confirm that mycotoxins are transported to aquatic systems by rain water through soil.


CD4(+)CD25(+) T Cells in primary malignant hypertension related kidney injury.
CD4(+)CD25(+) T cells are critical for maintenance of immunologic self-tolerance. We measured the number of CD4(+)CD25(+) cells in the patients with primary malignant hypertension related kidney injury, to explore the molecular pathogenesis of this disease. We selected 30 patients with primary malignant hypertension related kidney injury and 30 healthy volunteers. Information on clinical characteristics and laboratory tests was obtained from each subject. The number of CD4(+)CD25(+) cells and glomerular injury were assessed by flow cytometry and histopathology, respectively. Both serum IL-2, IL-4, and IL-6 and endothelial cell markers were analyzed by ELISA. ADAMTS13 antibody was detected by Western blotting. CD4(+)CD25(+) cells were significantly reduced in patients with primary malignant hypertension related kidney injury compared to controls (P < 0.05). The number of CD4(+)CD25(+) cells was negatively related to blood urea nitrogen, serum uric acid, proteinuria, and supernatant IL-4; whereas positively associated with estimated glomerular filtration rate in patients. Gradually decreasing CD4(+)CD25(+) cells were also found as increasing renal injury. Additionally, patients exhibited increasing supernatant IL-4, serum IL-2 and IL-6, endothelial cell markers, and anti-ADAMTS13 antibody compared with controls (all P < 0.05). CD4(+)CD25(+) cells may play a key role in the pathogenesis of primary malignant hypertension related kidney injury.


Use of established hepatoma cell lines in biotoxicology.
In conclusion, hepatoma cell lines displaying multiple combinations of bioactivating and inactivating enzymes, even if do not seem to represent liver-specific bio-indicators of toxicity, are a polyvalent liver system, suitable to evaluate particular end-points.


Intrapericardial echocardiography: a novel catheter-based approach to cardiac imaging.
Transvascular catheter-based intracardiac echocardiography has been successfully used to help guide catheter ablation and electrophysiologic procedures. It has recently been demonstrated that catheters can be safely placed into the pericardial space to allow for epicardial cardiac mapping and ablation. We evaluated the feasibility of catheter-based intrapericardial echocardiography (IPE) during such procedures to identify cardiac structures and visualize intracardiac catheters. IPE was performed in 7 goats by placing a phased-array ultrasound transducer contained within a 10F steerable catheter into the pericardial space using the same transthoracic subxyphoid approach as used to map and ablate epicardial ventricular tachycardia. Images were obtained of cardiac structures and of intracardiac ablation catheters. After the procedure, the hearts were harvested to assess for possible IPE-related lesions. The IPE catheter could be easily placed inside the pericardial space in all animals. In 7 of 7 cases, longitudinal and short-axis views of right- and left-sided chambers and valves were obtained, similar in orientation to transesophageal echocardiography. Visualization of atrial appendages (6/7), pulmonary veins (6/7), coronary arteries (6/7), and coronary sinus (3/6) was also feasible. Assessment of intracardiac transvalvar and venous blood flow was achieved by spectral and color Doppler. The ablation catheter could be clearly visualized inside cardiac chambers. No arrhythmias were induced with IPE catheter manipulation. After harvesting the hearts, no lesions resulting from the procedure were observed. In this experimental setting, IPE was able to provide detailed images of cardiac structures and establish the relative position of the ablation catheter.


Early closure of a multicenter randomized clinical trial of endoscopic stenting versus surgery for stage IV left-sided colorectal cancer.
The introduction of self-expandable metal stents has offered a promising alternative for palliation of malignant left-sided colonic obstruction. This randomized clinical trial aimed to assess whether a nonsurgical policy, with endoluminal stenting, is superior to surgical treatment in patients with stage IV left-sided colorectal cancer and imminent obstruction. Patients with incurable left-sided colorectal cancer who fulfilled the study criteria were randomly assigned to nonsurgical or surgical treatment. The primary outcome measure was survival in good health out of hospital (World Health Organization performance scores 0 or 1). A high number of serious adverse events in the nonsurgical arm led to premature closure of the trial. Ten patients were allocated to surgical treatment and 11 patients to nonsurgical palliation. The median survival in good health out of hospital during the first year was 56 days (interquartile range 7.5 - 338.5 days) in the surgical arm vs. 38 days (interquartile range 5.25 - 288.75 days) in the nonsurgical arm (P = 0.68). Eleven adverse events (six perforations) occurred in the nonsurgical arm vs. one adverse event in the surgical arm (P < 0.001). Of the six perforations, two were stent-related because they occurred at the proximal edge of the stent by erosion through a normal colon wall; one was probably stent-related (it was located in the region of the proximal half of the stent); one was a colon blowout; and two were late tumor perforations in patients on chemotherapy. The unexpected high rate of perforation in the nonsurgical arm might be specifically WallFlex-related or enteral stent-related in patients on chemotherapy and warrants attention.


Technical aspects of oxygen saving devices.
Oxygen economizing devices have been extensively studied, both at rest and during muscular exercise, in an attempt to increase the autonomy of a portable oxygen apparatus. The aim of this study is threefold: first, to suggest a simple method to verify in a simple way the technical accuracy of a demand flow oxygen delivery device; second, to suggest how we can monitor in a simple way the clinical efficacy of an economizer; and third, to remember that we can utilize an oxygen saving device to give a better protection than nasal prongs against the worsening of HbO2 desaturation induced by exercise.


Red blood cell transfusion and mortality effect in aneurysmal subarachnoid hemorrhage: a systematic review and meta-analysis protocol.
Aneurysmal subarachnoid hemorrhage (aSAH) is a devastating disease that leads to important morbidity and mortality in a young patient population. Anemia following aSAH is common and may be exacerbated by the treatments instituted by clinicians as part of standard care. The role and optimal thresholds for red blood cell (RBC) transfusion in this patient population remains unknown. We will conduct a systematic review of the literature using MEDLINE, EMBASE, and EBM Reviews (including Cochrane Central databases) using a comprehensive search strategy for observational and interventional studies of RBC transfusion in aSAH. Our primary objective is to evaluate the association of RBC transfusion with mortality in aSAH patients. Secondary objectives include a) determining associations between RBC transfusion and poor neurologic outcome, b) defining an optimal RBC transfusion threshold in aSAH patients, and c) describing complications associated with RBC transfusion in aSAH patients. We plan a descriptive reporting of all included citations including study characteristics, methodological quality, and reported outcomes. Clinical and statistical heterogeneity observed between studies will be described. If appropriate, meta-analyses of suitable studies and interpretation of their results will be performed. Effect measures will be converted to obtain relative risks and odds ratios (RR and ORs) with 95% confidence intervals and pooled according to study design (randomized trials and observational studies respectively) using a random effects model. This review will summarize the existing observational and trial evidence regarding RBC transfusion in aSAH patients. The analytical plan has made considerations for different study designs, both observational and interventional in nature, and will summarize the best available evidence to inform the end user and policy and guideline producers and to highlight areas in need of further study. PROSPERO CRD42014014806.


Rotational Atherectomy Combined with Drug Coated-Balloons for in-Stent Restenosis.
Refractory coronary in-stent restenosis remains a clinically relevant problem in interventional cardiology despite the use of drug coated balloon angioplasty and further drug eluting stent deployment. In this study, we investigated whether the novel approach of lesion debulking with rotational atherectomy prior to drug coated balloon angioplasty for challenging coronary in-stent restenosis is safe and effective. Procedural and registry data was retrospectively analysed for 26 patients who underwent rotational atherectomy immediately followed by drug coated balloon angioplasty to 43 coronary in-stent restenosis lesions with mean follow up of 19 months. Lesion success was achieved in all cases with no major procedural complications. There were no instances of death or myocardial infarction in the follow up period. Target lesion revascularisation occurred in six patients and target vessel revascularisation occurred in eight patients. All target lesion revascularisation occurred in lesions that had already failed drug coated balloon angioplasty without debulking previously while four such lesions were free of lesion failure in the follow up period. Lesion debulking with rotational atherectomy followed by drug coated balloon angioplasty is a feasible treatment option for selected cases of in-stent restenosis. Further study is needed to fully assess its efficacy in comparison to conventional treatment.


Facilitating nursing students' skill training in distance education via online game-based learning with the watch-summarize-question approach during the COVID-19 pandemic: A quasi-experimental study.
Key challenges for clinical education during the COVID-19 pandemic include how to continue teaching and learning, how to teach core clinical skills, and how to demonstrate professional and practical skills in various clinical situations. Therefore, nursing students need to learn how to assist with in-patient intubation, eliminate accumulated sputum overflow, and the basic techniques of sputum suction. We proposed and investigated an approach to integrating online game-based learning with the watch-summarize-question strategy to improve nursing students' learning achievement, self-efficacy, learning engagement, and learning satisfaction in sputum suction skill training. A quasi-experimental study with pretest and posttest design. We randomly allocated 45 first-year nursing students to an experimental group (n = 21) or a control group (n = 24) at a school of nursing in a university. The experimental group adopted the online game-based learning and watch-summarize-question strategy, while the control group used video-based learning. Participants were assessed on learning achievement of sputum suction skills, self-efficacy, learning engagement, and learning satisfaction before and after the intervention. The experimental group, which used the proposed approach, achieved statistically significant higher learning achievement, self-efficacy, learning engagement, and learning satisfaction than the control group. The integration of online game-based learning with the watch-summarize-question strategy demonstrated a positive impact on nursing students' sputum suction skill training. Nurse educators and researchers should consider integrating computer technology and teaching strategies to facilitate nursing education.


Proliferation of T8-positive cytolytic T lymphocytes in response to thyroglobulin in human autoimmune thyroiditis: analysis of cell interactions and culture requirements.
These experiments were designed to analyze the involvement of T-lymphocyte subpopulations in autoimmune thyroid disorders such as Graves' Disease (GD) and Hashimoto's Disease (HD). In a first set of experiments, lymphocytes isolated from thyroid infiltrates or from peripheral blood of GD and HD patients were analyzed for the expression of various surface antigens. While HLA-DR + T cells were numerous among thyroid infiltrating T lymphocytes in both groups of patients, the proportions of T8 + cells (as defined by their reactivity with the B 9.4 monoclonal antibody specific for T8 surface molecule) were strikingly different in HD and GD. In the latter group of patients only 19% of infiltrating T cells were T8 +, whereas these cells represented approximately 50% in four HD patients. Given the previous demonstration that all T cells expressing T8 antigen are cytolytic T lymphocytes (CTL) or their precursors (CTL-P) in conjunction with the fact that lymphocytes from HD or GD patients are known to proliferate in vitro in response to human tg (Htg), we further analyzed the T-cell subset(s) responsible for in vitro proliferation to Htg. In these experiments, peripheral blood T lymphocytes purified from patients with GD or HD were cultured with 1 microgram/ml Htg and irradiated autologous T-depleted mononuclear cells as the source of antigen presenting cells (APC). The proportions of T8 + cells declined considerably during culture in GD patients, but at Days 6 to 9, T8 + cells represented as much as 51% of cultured T lymphocytes from patients with HD. Moreover, the majority of T8 + cells were medium-large size lymphoblasts. Removal of Htg at Day 6 resulted in both abrogation of proliferative responsiveness and in decreases of T8 + percentages. Further analysis of the cell interactions leading to T8 + cell proliferation in response to Htg showed that helper/inducer T cells, as defined by 5/9 antigen expression, were strictly required. Collectively, these features are reminiscent of the T-cell involvement in experimental autoimmune thyroiditis of mice and stress for the first time the potential role of CTL in tissue damage occurring in Hashimoto's thyroiditis.


Asynchronous ventricular relaxation: an angiographic temporal analysis of asynchronous left ventricular relaxation in man.
Segmental "early relaxation" is a common angiographic finding. An attempt was made to elucidate the temporal characteristics of this event. Twenty subjects with and 20 without segmental early relaxation were studied. The left ventricular diastolic relaxation time was precisely determined angiographically, and the findings were as follows: 0.12 +/- 0.04 second (mean +/- standard deviation) in the normal patients, 0.14 +/- 0.03 second in the patients with coronary artery disease but no segmental early relaxation and 0.20 +/- 0.04 second in the patients with segmental early relaxation. These findings indicate that early relaxation is associated with a significantly prolonged ventricular relaxation time. The use of the term asynchronous ventricular relaxation is proposed to denote the disturbed diastolic properties of the ventricle with "early relaxation."


Necrotizing pneumonia in cats associated with infection by EF-4a bacteria.
Two cases of severe necrotizing pneumonia in cats caused by Gram-negative bacteria called "Eugonic Fermenter 4a" (EF-4a) are described. The gross lesions resembled to those commonly seen with lung tuberculosis in other animal species.


IL-37b suppresses epithelial mesenchymal transition in hepatocellular carcinoma by inhibiting IL-6/STAT3 signaling.
Interleukin-37b (IL-37b), a vital negative regulator of the innate immune system, has been reported to be a tumor inhibitor in different type of cancers. However, little is known about the relationship between IL-37b and hepatocellular carcinoma (HCC). The present study aimed to investigate the potential roles of IL-37b in HCC progression. Subjects (n = 237) were recruited, and serum IL-37b was measured using ELISA. The tumor-suppressive capacity and underlying mechanisms of IL-37b in HCC were investigated in vitro and in vivo. Compared to healthy controls, serum IL-37b levels were elevated in chronic hepatitis B (CHB) patients but decreased significantly in HBV-HCC patients, especially for those with portal venous tumor thrombus. Low level serum IL-37b in HBV-HCC patients correlated with high HCC stage and poor overall survival and disease-free survival. In vitro and in vivo, recombinant human IL-37b inhibited proliferation and metastasis in HCC cells. Furthermore, IL-37b inhibited epithelial mesenchymal transition in HCC cells in vitro by downregulating IL-6, pSTAT3 (Y705), N-cadherin, and vimentin expression and by upregulating E-cadherin expression. These effects were partially reversed by transfection of adenovirus encoding human IL-6. IL-37b inhibits HCC growth, metastasis and epithelial mesenchymal transition by regulating IL-6/STAT3 signaling. Serum IL-37b may be a biomarker for HBV-HCC and its staging.


Finding time.
We understand time through our models of it. These are typically models of our physical chronometers, which we then project into our subjects. A few of these models of the nature of time and its effects on the behavior of organisms are reviewed. New models, such as thermodynamics and spectral decomposition, are recommended for the potential insights that they afford. In all cases, associations are essential features of timing. To make them, time must be discretized by stimuli such as hours, minutes, conditioned stimuli, trials, and contexts in general. Any one association is seldom completely dominant, but rather shares control through proximity in a multidimensional space, important dimensions of which may include physical space and time as rendered by Fourier transforms.


Chuanminshen violaceum polysaccharides improve the immune responses of foot-and-mouth disease vaccine in mice.
Water-soluble polysaccharides from Chuanminshen violaceum (CVPS) were evaluated for their physicochemical properties, monosaccharide composition, and adjuvant potential to specific cellular and humoral immune responses in a mouse model of foot-and-mouth disease virus (FMDV) vaccination. The average molecular weight (Mw) of the CVPS was 968.31 kDa. The monosaccharide components of the CVPS was rhamnose, arabinose, fucose, mannose, glucose, and galactose with a relative mass of 6.29%, 21.87%, 16.59%, 12.54%, 13.07%, and 28.05%, respectively. Administering CVPS as an adjuvant significantly enhanced the phagocytic capacity of peritoneal macrophages, splenocyte proliferation, and the activity of NK cells and CTL as well as increased FMDV-specific IgG and IgG subclass antibody titers. Moreover, CVPS increased the expression of IL-2, IFN-γ, and IL-4 in CD4(+) T cells and IFN-γ expression in CD8(+) T cells. Additionally, CVPS enhanced CD40(+), CD80(+), and CD86(+) expression on DCs. Moreover, CVPS upregulated MHC-I/II, TLR-2/4 mRNA levels. In contrast, CVPS downregulated TGF-β mRNA expression and the frequency of CD4(+)CD25(+)Foxp3(+) Treg cells. Taken together, these results indicate that administering CVPS as an adjuvant enhances both cellular and humoral immune responses via the TLR-2 and TLR-4 signalling pathways, thereby promoting DC maturation and suppressing TGF-β expression and Treg frequency.


Pathogenic compound heterozygous ATP7B mutations with hypoceruloplasminaemia without clinical features of Wilson's disease.
The authors report a 44-year-old man with a history of attention deficit and hyperactivity disorder, obsessive compulsive behaviour, vocal tics, depression, and anxiety, in whom a compound heterozygous ATP7B mutation was found, associated with hypoceruloplasminemia, but without clinical or pathological manifestation of Wilson's disease (WD). Genetic testing revealed a compound heterozygous ATP7B mutation already described in WD, p.Met645Arg (C1934TG/c.51+4A→T). Hypoceruloplasminaemia was detected but no clinical manifestations (hepatic or central nervous system) of WD were present. The authors conclude that patients can carry a heterozygous mutation of the ATP7B gene that is associated with hypoceruloplasminaemia and display no overt clinical hepatic and/or central nervous system manifestations of WD.


Interleukin-6 serum levels correlate with footpad swelling in adjuvant-induced arthritic Lewis rats treated with cyclosporin A or indomethacin.
To investigate the role of interleukin 6 (IL-6) in adjuvant-induced arthritis, serum from adjuvant-immunized Lewis rats treated with cyclosporin, indomethacin, or saline was evaluated for IL-6 activity. Inflammation was quantitated by measuring paw volume. We found that an increase in serum IL-6 activity parallels the kinetics of paw edema development in adjuvant-immunized rats. Daily treatment with 5 mg cyclosporin A/kg prevented the increase in paw volume and held serum IL-6 activity to levels observed in untreated (normal) rats. Daily treatment with 1 mg indomethacin/kg resulted in a 50% reduction in serum IL-6 levels and a significant decrease (approximately 50%) in paw volume on Day 17 compared to saline-treated rats. Linear regression analysis confirmed the positive correlation between mean paw volume and mean serum IL-6 activity (R2 = 0.783, P less than 0.01 on Day 17) in normal, arthritic, and cyclosporin A- or indomethacin-treated groups. These results are consistent with a role for IL-6 in the pathology of arthritis and suggest that serum IL-6 activity may be a useful parameter for monitoring disease activity.


Multicomponent multicatalyst reactions (MC)2R: one-pot synthesis of 3,4-dihydroquinolinones.
A Rh/Pd/Cu catalyst system led to an efficient synthesis of dihydroquinolinones in one-pot, two operations. The reaction features the first triple metal-catalyzed transformations in one reaction vessel, without any intermediate workup. The conjugate-addition/amidation/amidation reaction sequence is highly modular, divergent, and practical.