Factors affecting pregnancy outcome resulting from assisted reproductive technology (ART).
A retrospective, observational study of pregnancy outcome was performed on variables maintained in an ART database to determine factors that might affect miscarriage rate in pregnancies resulting from assisted reproduction technologies (ART). Previously infertile couples, where conception was achieved after ART, were included. Seven hundred and ninety-four consecutive clinical pregnancies, diagnosed by ultrasound documentation of the gestation sac in the first trimester were divided into 2 groups: 'miscarriage' and 'term birth'. Differences between the groups were analysed using crosstable regression analyses or t-test in second yearly cohorts. A statistically significant positive relationship was seen between age and spontaneous abortion rate (p = 0.008) with a major increase after the age of 38 years. There was no significant difference in the mean number of oocytes retrieved between groups (p = 0.17). While there was a significant negative correlation between maternal age and the total number of oocytes collected (p < 0.001), there was no statistical difference between those women who miscarried or delivered a live infant. No relationships were found with any other variables analysed. Maternal age is probably the most important factor in pregnancy outcome in ART. This survey could not determine any other variables as being major prognostic determinant for miscarriage once pregnancy was attained.


Susac's Syndrome--update.
Susac's Syndrome (SS) consists of the clinical triad of encephalopathy, branch retinal artery occlusion (BRAO), and hearing loss (HL). It is an autoimmune endotheliopathy affecting the precapillary arterioles of the brain, retina, and inner ear (cochlea and semicircular canals). The age range extends from 7 to 72 years, but young women (20-40) are most vulnerable. Headache routinely accompanies the encephalopathy and may be constant (best explained by leptomeningeal involvement), migrainous, or both. Multifocal neurological manifestations--particularly bilateral long-tract signs--commonly accompany the encephalopathy, which is laden with psychiatric features, confusion, memory loss and other cognitive changes. Left untreated, dementia can ensue. SS has an unexplained proclivity for attacking the central corpus callosum. In its encephalopathic form, pathognomonic callosal lesions permit an immediate diagnosis. We believe that the diagnosis of SS can be made when only the encephalopathy and pathognomonic MRI lesions are present; the BRAO and HL need not be present. We have also found the "string of pearls" MRI finding--the studding of the internal capsules with microinfarcts--to be most helpful--if not pathognomonic. This sign is always associated with the clusters of corpus callosum lesions, is especially striking on diffusion weighted imaging, and is associated with long-tract findings. We discuss the newly appreciated BRAO subset of SS and offer preliminary treatment suggestions for this subset. We also call attention to our development of an International Collaborative Study of SS and an educational website (http://www.ucalgary.ca/susac).


Movement disorder caused by FRRS1L deficiency may be associated with morphological and functional disorders in Purkinje cells.
Ferric Chelate Reductase 1 Like (FRRS1L) protein has been identified as an auxiliary regulatory protein for the α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPAR). FRRS1L is highly expressed in the cerebellum and other brain regions associated with the control of motor function. Loss of FRRS1L has been shown to lead to impaired synaptic transmission via AMPARs and to movement disorders. We found that deletion of the FRRS1L gene causes hyperactivity, reduced muscle strength, impaired coordination, and ataxia in mice. Deletion also impairs Purkinje cell dendritic spine formation and AMPAR expression in the cerebellum and damages the electrophysiological discharge rhythm of Purkinje cells. Cerebrospinal fluid examination and oleic acid (OA)-induced lipid accumulation monitoring in FRRS1L-knockdown SH-SY5Y cells indicated that FRRS1L deficiency could lead to aberrant metabolism of amino acids, glucose, and lipids. In summary, we found that the deletion of FRRS1L leads to impaired motor coordination and cerebellar ataxia in mice, which might be related to the reduced expression of AMPARs, metabolic deviations, and dysplastic functional defects in Purkinje cells.


Cell selection for increasing resistance of ornamental plants to copper.
Cell selection was used to obtain copper-resistant plants. Developed technologies for obtaining copper-resistant plants Agrostis stolonifera and Chrysanthemum carinatum can be applied to other plant species. We obtained copper-resistant plants Festuca rubra, Brachycome iberidifolia and Linum perenne. The concept of obtaining plants resistant to copper has been developed. This concept consists of two methods. The first method is applicable when calli is highly sensitive to copper. The second method is applicable when calli are moderately sensitive to copper.


A modified perineal approach to reconstruction of membranous urethra for stricture.
Difficulty in obtaining perineal exposure of the membranous urethra in cases of traumatic stricture led to the development of the combined perineosuprapubic and transpubic approaches. Use of the standard perineal incision for perineal prostatectomy with division of the urogenital diaphragm in the midline from behind up to and encircling the urethra, and temporary suturing of the flaps of the urogenital diaphragm that are created to the lateral edges of the skin incision provides good exposure of the membranous and proximal bulbous urethra, thus, making reconstruction easier. This approach should minimize the need for the transpubic or perineosuprapubic approaches. It has been used for urethroplasty in 12 patients with traumatic membranous strictures, 5 with traumatic bulbomembranous strictures, 7 with post-gonococcal strictures involving the proximal bulbous and distal membranous urethra, and 1 with a bulbomembranous urethral rupture.


Thiolated polymeric hydrogels for biomedical application: Cross-linking mechanisms.
This review focuses on the synthesis of hydrogel networks using thiomers such as thiolated hyaluronic acid, chitosan, cyclodextrin, poly(ethylene glycol) and dextran that are cross-linked via their thiol substructures. Thiomers have been widely investigated as matrix of hydrogels due to the high reactivity of these sulfhydryl moieties. They are well known for their in situ gelling properties due to the formation of inter- and intra-chain disulfide bonds. Furthermore, as thiol groups on the polymeric backbone of thiomers cannot only react with each other but also with different other functional groups, several "click" methods such as thiol-ene/yne, Michael type addition and thiol-epoxy reactions have been developed within the last decades to fabricate thiomer hydrogels. These hydrogels are meanwhile used as scaffolds for tissue engineering, regenerative medicine, diagnostics and as matrix for drug and protein delivery.


Physiologically-Based Pharmacokinetic Modeling for Drug Dosing in Pediatric Patients: A Tutorial for a Pragmatic Approach in Clinical Care.
It is well-accepted that off-label drug dosing recommendations for pediatric patients should be based on the best available evidence. However, the available traditional evidence is often low. To bridge this gap, physiologically-based pharmacokinetic (PBPK) modeling is a scientifically well-founded tool that can be used to enable model-informed dosing (MID) recommendations in children in clinical practice. In this tutorial, we provide a pragmatic, PBPK-based pediatric modeling workflow. For this approach to be successfully implemented in pediatric clinical practice, a thorough understanding of the model assumptions and limitations is required. More importantly, careful evaluation of an MID approach within the context of overall benefits and the potential risks is crucial. The tutorial is aimed to help modelers, researchers, and clinicians, to effectively use PBPK simulations to support pediatric drug dosing.


Thrombolysis as first-line therapy for Medtronic/HeartWare HVAD left ventricular assist device thrombosis.
We reviewed our institutional experience with intravenous thrombolysis (TL) as first-line therapy in patients with Medtronic/HeartWare HVAD left ventricular assist device pump thrombosis (PT). From March 2006 to November 2018, 30 Medtronic/HeartWare HVAD left ventricular assist device patients had 48 PT events. We analysed outcomes with intravenous Alteplase as a first-line therapy for PT. Pump exchange or urgent heart transplantation was only considered after the failure of TL or existing contraindications to TL. TL was used as the first-line therapy in 44 PT events in 28 patients without a contraindication to TL. TL was successful in 61.4% of PT events. More than 1 cycle of TL was necessary in 55.6% of events. The combined success of TL and heart transplantation or device exchange was 81.8%. In 15.9% of events, PT was fatal. Causes of death were severe complications (9.1%) related to TL or discontinuation of therapy for multi-organ failure (6.8%). Intracranial bleeding and arterial thromboembolism were observed in 4.5% and 11.5% of the PT events after TL. Intravenous TL as a first-line therapy for PT in Medtronic/HeartWare HVAD patients can be a reasonable treatment option and does not preclude subsequent heart transplantation or device exchange. However, thromboembolic and bleeding complications are common. The decision to perform TL or device exchange should, therefore, be made on an individual basis after balancing the risks and benefits of different treatment approaches.


Molecular Editing Enhances Oxidation Resistance of Menaquinone-Targeting Antibiotics Lysocin E and WAP-8294A2.
Lysocin E (1 a) and WAP-8294A2 (2 a) are peptidic natural products with 37- and 40-membered macrocycles, respectively. Compounds 1 a and 2 a have potent antibacterial activities against Gram-positive bacteria and share a unique mode of action. The electron-rich indole ring of d-Trp-10 of 1 a and 2 a interacts with the electron-deficient benzoquinone ring of menaquinone, which is a co-enzyme in the bacterial respiratory chain. Formation of the electron-donor-acceptor complex causes membrane disruption, leading to cell death. Despite the promising activities of 1 a and 2 a, the susceptibility of Trp-10 to oxidative degradation potentially deters the development of these compounds as antibacterial drugs. To address this issue, we replaced the indole ring with more oxidation-resistant aromatics having a similar shape and electron-rich character. Specifically, analogues with benzofuran (1 b/2 b), benzothiophene (1 c/2 c), and 1-naphthalene (1 d/2 d) rings were designed, and chemically prepared by full solid-phase total syntheses. Antibacterial assays of the six analogues revealed similar activities of 1 d/2 d and markedly reduced activities of 1 b/2 b and 1 c/2 c compared with 1 a/2 a. Equipotent 1 d and 2 d both showed high resistance to oxidation by peroxyl radicals. Hence, the present study demonstrates a new molecular editing strategy for conferring oxidation stability on natural products with pharmacologically useful functions.


Assessment of coronary flow velocity reserve with phase-contrast cine magnetic resonance imaging in patients with heavy coronary calcification.
Coronary flow velocity reserve (CFVR) can be noninvasively measured by phase-contrast cine magnetic resonance imaging (PC-MRI). Heavy coronary calcification degrades the diagnostic accuracy for the detection of coronary arterial stenosis on computed tomography (CT). The aim of this study was to evaluate the value of CFVR measurement with PC-MRI for detecting significant coronary stenoses in patients with heavy coronary calcification. Sixteen patients (71 ± 8 years) with coronary calcium score above 400 who had suspected moderate coronary stenosis (50-69% diameter stenosis) on CT angiography were prospectively studied. The CFVR values, calculated as the ratio of peak flow velocity during hyperemia to the peak flow velocity at rest, were measured using breath-hold PC-MRI with 3 T system, and were compared with the results of quantitative coronary angiography (QCA). The mean coronary calcium score was 985 ± 378. CFVR was successfully determined with PC-MRI in 17/18 (94%) vessels. Using a threshold of 1.4 for CFVR, the sensitivity, specificity, and positive and negative predictive value for detecting ≥ 50% stenosis on QCA was 88% (7/8), 89% (8/9), 88% (7/8), 89% (8/9), respectively. When MRI CFVR measurements was added to CT angiography for the evaluation of coronary stenosis, the positive predictive value was 88% (7/8), while the positive predictive value of CT angiography alone was 44% (8/18). PC-MRI can provide noninvasive detection of altered CFVR caused by significant stenosis in patient. CFVR measurement by PC-MRI is useful for diagnosing physiologically significant coronary stenosis in patients with high calcium score on CT.


Bricolage as an expression of self and of cultural and familial foodways among people living in prison-'You make what you can with anything you can get'.
In this article, we use Lévi-Strauss's (1962, The savage mind, University of Chicago Press) concept of 'bricolage' to explore informal food preparation among men in Scottish prisons. The art of 'making do with whatever is at hand', in innovative and creative ways, to give new functions to everyday items has recently been reimagined and applied to the field of food. It has been used to explore the practice of informal food networks in resource poor environments; investigate how small food businesses come up with new and innovative recipes ideas; and study the way Michelin-starred chefs responded to the COVID-19 pandemic through philanthropic activities. Our aim is to use bricolage as a lens through which to answer questions about whether more autonomy over food might contribute to overall health and wellbeing in prisons. Drawing on in-depth empirical data from qualitative interviews with 20 men in Scottish prisons, we explore how bricolage is used to escape the monotony of prison-issued meals and the tedium of the prison regime; counter threats to self and identity; create and maintain social relationships through joint enterprise and commensality; and create culinary experiences that afford a sense of control and normality in an environment synonymous with 'spoiled identity' (Goffman, 1961, Asylums: Essays on the social situation of mental patients and other inmates, Penguin.).


CGP 6809, a sugar-containing nitrosourea derivative: pharmacological and physicochemical properties.
CGP 6809 is a water-soluble nitrosourea derivative with quite distinct chemical and biological properties as compared with the well-known representatives of this class of compounds. It is related to the antibiotic streptozotocin, from which it is distinguished in the structure of the sugar moiety and the position of the methylnitrosourea residue. CGP 6809 possesses practically the same alkylating potential as streptozotocin; however, its carbamoylating activity is comparable with that of CCNU. In contrast to other nitrosourea derivatives, CGP 6809 showed relatively little activity in murine leukemias but was markedly active in solid transplantable melanomas (Harding-Passay, B16), in the 11095 prostate carcinoma, and in a substrain of Yoshida hepatoma (AH 7974) resistant to BCNU and CCNU. In the Ehrlich and Yoshida ascitic tumors complete responses were seen with no toxic death. Dose-dependent activity was found in the human lung carcinoma MBA 9812 and almost complete growth inhibition was achieved in the human melanoma WM 47 by both the oral and parenteral routes of administration. However, mammary tumor lines (Ca 755, 2661/61, R-3230AC), the Guerin-T8 uterus epithelioma, and the Rous sarcoma/S-R proved to be relatively refractory to this drug. This was also the case for the Lewis lung carcinoma implanted i.m. or s.c. However, development of lung metastases was markedly inhibited. Combination therapy using CGP 6809 with cyclophosphamide, 5-fluorouracil, or chlorambucil in the same model led to partial responses of the primary tumor as well as almost total eradication of lung metastases.


Local spectroscopy of a gate-switchable moiré quantum anomalous Hall insulator.
In recent years, correlated insulating states, unconventional superconductivity, and topologically non-trivial phases have all been observed in several moiré heterostructures. However, understanding of the physical mechanisms behind these phenomena is hampered by the lack of local electronic structure data. Here, we use scanning tunnelling microscopy and spectroscopy to demonstrate how the interplay between correlation, topology, and local atomic structure determines the behaviour of electron-doped twisted monolayer-bilayer graphene. Through gate- and magnetic field-dependent measurements, we observe local spectroscopic signatures indicating a quantum anomalous Hall insulating state with a total Chern number of ±2 at a doping level of three electrons per moiré unit cell. We show that the sign of the Chern number and associated magnetism can be electrostatically switched only over a limited range of twist angle and sample hetero-strain values. This results from a competition between the orbital magnetization of filled bulk bands and chiral edge states, which is sensitive to strain-induced distortions in the moiré superlattice.


High SYT7 expression is associated with poor prognosis in human non-small cell lung carcinoma.
Synaptotagmin 7 (SYT7) can encode a single-pass 46-kDa transmembrane protein which located on human chromosome 11q12.2. It has been reported to be dysregulated in several cancers; however, there are few reports on the role of SYT7 in non-small cell lung carcinoma (NSCLC). The purpose of our study was to investigate the expression of SYT7 in NSCLC and its relationship with the prognosis of NSCLC. Differences in SYT7 expression were explored by using a public database and tissue samples. The prognostic value of SYT7 and its expression correlation with clinical parameters were evaluated by statistical analysis. Our current study found that elevated mRNA and protein levels of SYT7 in NSCLC tissues compared to adjacent normal tissues. The high expression of SYT7 in NSCLC patients was positively correlated with tumour differentiation (P = 0.031) and pT (P = 0.041). The higher SYT7 expression had a shorter survival time than those with lower SYT7 expression in NSCLC patients. Furthermore, multivariate analysis demonstrated that the expression of SYT7 was an unfavourable independent prognostic factor for NSCLC (P = 0.044). In conclusion, SYT7 was upregulated in NSCLC tissues and maybe a prognostic and diagnostic factor of NSCLC.


Dual-Function Fibrous Co-Polypeptide Scaffolds for Neural Tissue Engineering.
This paper reports dual-function (high cell attachment and cell viability) fibrous scaffolds featuring aligned fibers, displaying good biocompatibility and no cytotoxicity. These scaffolds are fabricated through the electrospinning of a co-polypeptide comprising molar equivalents of N6 -carbobenzyloxy-l-lysine and γ-benzyl-l-glutamate, with the lysine moieties enhancing cell adhesion and the neural-stimulating glutamate moieties improving cell viability. These new scaffolds allow neural cells to attach and grow effectively without any special surface treatment or coating. Pheochromocytoma (PC-12) cells grown on these scaffolds exhibit better neuronal activity and longer neurite length, relative to those grown on scaffolds prepared from their respective homo-polypeptides. When the scaffolds are partially hydrolyzed such that they present net positive charge and increased hydrophilicity, the cell viability and neurite growth both increase further. Accordingly, these novel co-polypeptide fibrous scaffolds have potential applications in neural tissue engineering.


PRDM15 interacts with DNA-PK-Ku complex to promote radioresistance in rectal cancer by facilitating DNA damage repair.
Neoadjuvant radiotherapy is a standard treatment for locally advanced rectal cancer, however, resistance to chemoradiotherapy is one of the main obstacles to improving treatment outcomes. The goal of this study was to explore the role of PRDM15 involved in the radioresistance of colorectal cancer and to clarify the underlying mechanism. In present study, we demonstrated that, after DNA damage, PRDM15 was upregulated and localized to DNA damage sites, co-localizing with γ-H2AX. Knockdown of PRDM15 inhibited DNA damage repair and increased radiosensitivity in colorectal cancer cells. Mechanistically, PRDM15 promoted DNA repair by interacting with DNA-PKcs and Ku70/Ku80 complex. In preclinical models of rectal cancer, knockdown of PRDM15 sensitized cell derived xenograft and patient derived xenograft to radiotherapy. In 80 rectal cancer patients treated with neoadjuvant chemoradiotherapy, higher PRDM15 expression was observed associated with weaker tumor regression and poorer prognosis. Our findings revealed that inhibiting PRDM15 was potent to overcome radioresistance through abrogating DNA repair in colorectal cancer cells. Additionally, the expression level of PRDM15 could be applied to predict radiotherapy responsiveness and the outcome of neoadjuvant radiotherapy in rectal cancer patients.


Pharmacological and genetic evidence indicates that combined inhibition of NR2A and NR2B subunit containing NMDA receptors is required to disrupt prepulse inhibition.
Glutamate signalling through the N-methyl-D-aspartate (NMDA) receptor is of critical importance for normal central nervous system (CNS) function, as indicated by the marked behavioural disturbances produced by non-subtype selective NMDA antagonists such as dizocilpine (MK-801). The present studies were designed to investigate the involvement of the two major NMDA receptor subunits in the central nervous system, i.e. NR2A and NR2B, on sensorimotor gating in mice. These experiments utilised the non-subtype-selective NMDA antagonist dizocilpine, a line of NR2A-KO mice and the selective NR2B antagonist Ro 63-1908, in the study of pre-pulse inhibition of the startle response (PPI). The non-selective NMDA receptor antagonist dizocilpine (0.1-1 mg/kg, IP) robustly disrupted PPI in wild-type mice. Conversely, selective genetic or pharmacological inhibition of either the NMDA NR2A or NR2B receptor subunit containing receptors, respectively, had no effect on PPI. Thus, NR2A KO mice showed normal PPI compared with wild-type littermate controls, and administration of Ro 63-1908 (1-10 mg/kg IP) to wild-type mice did not affect PPI. However, selective inhibition of NR2A and NR2B by administration of Ro 63-1908 to NR2A KO mice significantly disrupted PPI. These data imply that concomitant inhibition of both NR2A and NR2B subunit-containing NMDA receptors is necessary to disrupt PPI, suggesting that inhibition of NR2A and NR2B-containing NMDA receptors is required to elicit behaviours suggestive of psychomimetic effects in man.


Varicella Zoster Virus Vasculopathy; An HIV adult presenting with multiple strokes.
VZV vasculopathy has been associated with granulomatous vasculitis where there is vessel wall damage and transmural inflammation. VZV vasculopathy has been previously called granulomatous angiitis, VZV vasculitis or post-varicella arteriopathy. Intracerebral VZV vasculopathy can occur in children and adults, either after primary infection or after viral reactivation. Where-as varicella primary infection is a common cause of stroke in children, in adults there is an increased risk of stroke after herpes zoster. Here we present a 38-year-old immunocompromised patient who presents to us with multiple cerebral infarcts post primary infection and imaging showing distribution similar to those in children.


Trends in Stroke Incidence and 28-Day Case Fatality in a Nationwide Stroke Registry of a Multiethnic Asian Population.
This study investigated trends in stroke incidence and case fatality overall and according to sex, age, ethnicity, and stroke subtype in a multiethnic Asian population. The Singapore Stroke Registry identifies all stroke cases in all public hospitals using medical claims, hospital discharge summaries, and death registry data. Age-standardized incidence rates and 28-day case-fatality rates were calculated for individuals aged ≥15 years between 2006 and 2012. To estimate the annual percentage change of the rates, a linear regression model was fitted to the log rates, and a Wald test was performed to test for trend. P values <0.05 were considered significant. A total of 40 623 cases were recorded. The total stroke incidence fell by ≈12.0%, and case fatality fell by 17.2% in the study. Declining trends in stroke incidence were stronger in women (female: -2.94; 95% confidence interval [CI], -3.43 to -2.44; male: -1.80; 95% CI, -2.58 to -1.02); in the older age groups (≥65 years: -3.62; 95% CI, -4.30 to -2.94; 50-64 years: -1.26; 95% CI, -1.97 to -0.55; <50 years: 3.33; 95% CI, 1.49 to 5.20), in Chinese (-2.64; 95% CI, -3.15 to -2.13), Indians (-3.78; 95% CI, -5.93 to -1.58), and others (-12.73; 95% CI, -18.93 to -6.06) compared with Malays (2.58; 95% CI, 1.17 to 4.02); and in ischemic stroke subtype (ischemic: -2.43; 95% CI, -3.13 to -1.73; hemorrhagic: -1.02; 95% CI, -2.04 to 0.01). Subgroup-specific findings for case fatality were similar. This is the first countrywide hospital-based registry study in a multiethnic Asian population, and it revealed marked overall reductions in stroke incidence and case fatality. However, it also identified important population groups with less favorable trends, especially younger adults and those of Malay ethnicity.


A review of Nipah and Hendra viruses with an historical aside.
The emergence of Hendra and Nipah viruses in the 1990s has been followed by the further emergence of these viruses in the tropical Old World. The history and current knowledge of the disease, the viruses and their epidemiology is reviewed in this article. A historical aside summarizes the role that Dr. Brian W.J. Mahy played at critical junctures in the early stories of these viruses.


The RESOLVE Trial for people with chronic low back pain: statistical analysis plan.
Statistical analysis plans describe the planned data management and analysis for clinical trials. This supports transparent reporting and interpretation of clinical trial results. This paper reports the statistical analysis plan for the RESOLVE clinical trial. The RESOLVE trial assigned participants with chronic low back pain to graded sensory-motor precision training or sham-control. We report the planned data management and analysis for the primary and secondary outcomes. The primary outcome is pain intensity at 18-weeks post randomization. We will use mixed-effects models to analyze the primary and secondary outcomes by intention-to-treat. We will report adverse effects in full. We also describe analyses if there is non-adherence to the interventions, data management procedures, and our planned reporting of results. This statistical analysis plan will minimize the potential for bias in the analysis and reporting of results from the RESOLVE trial. ACTRN12615000610538 (https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=368619).


Investigation of the subtype of alpha 2-adrenoceptor mediating prejunctional inhibition of cardioacceleration in the pithed rat heart.
1. We have investigated the subtype of alpha 2-adrenoceptor mediating prejunctional inhibition of cardioacceleration in the pithed rat heart in comparison with alpha 2-adrenoceptor ligand binding sites. 2. In pithed rats, prejunctional alpha 2-adrenoceptors were investigated in terms of the ability of alpha 2-adrenoceptor antagonists to shift the inhibitory potency of the alpha 2-adrenoceptor agonist, xylazine, against the tachycardia to a single electrical stimulus given via the pithing rod. 3. Antagonist potency at prejunctional alpha 2-adrenoceptors in pithed rat heart was correlated with antagonist affinity at alpha 2-adrenoceptor ligand binding sites in membranes of rat kidney and submandibular gland labelled with [3H]-yohimbine. 4. The correlation with the prejunctional alpha 2-adrenoceptor in pithed rat heart was best for the alpha 2D-adrenoceptor ligand binding site of rat submandibular gland (r = 0.98, n = 10, P < 0.0001), as compared to correlations with the alpha 2A-adrenoceptor ligand binding site of human platelet (r = 0.90, n = 9, P < 0.001), the alpha 2B-adrenoceptor ligand binding site of rat kidney (r = 0.82, n = 10, P < 0.01) and with published results for the alpha 2C-adrenoceptor ligand binding site (r = 0.48, n = 6, NS). 5. It is concluded that the functional prejunctional alpha 2-adrenoceptor of pithed rat heart closely resembles the alpha 2D-adrenoceptor ligand binding site of rat submandibular gland.


Role of adenine phosphoribosyltransferase in adenine uptake in wild-type and APRT- mutants of CHO.
Adenine uptake in cultured Chinese hamster fibroblasts showed biphasic saturation kinetics. The transport system was highly specific for adenine and was competitively inhibited by adenosine. Utilizing mutant clones of Chinese hamster fibroblasts that have either reduced or negligible adenine phosphoribosyltransferase (APRT) activity, we found that (1) adenine was not accumulated against a concentration gradient in the absence of APRT activity and (2) after rapid initial uptake equal to that of the parent the rates of adenine accumulation found for the mutants correlated strongly with their residual APRT activities. Furthermore, using either artificially depressed phosphoribosylpyrophosphate pool size and APRT activities or the mutants with decreased APRT activity, we found that adenine transport was independent of phosphorylation by APRT. These studies suggest that adenine is transported as the free base by facilitated diffusion and is subsequently phosphorylated by APRT.


Sexual Signals Persist over Deep Time: Ancient Co-option of Bioluminescence for Courtship Displays in Cypridinid Ostracods.
Although the diversity, beauty, and intricacy of sexually selected courtship displays command the attention of evolutionists, the longevity of these traits in deep time is poorly understood. Population-based theory suggests sexual selection could either lower or raise extinction risk, resulting in high or low persistence of lineages with sexually selected traits. Furthermore, empirical studies that directly estimate the longevity of sexually selected traits are uncommon. Sexually selected signals-including bioluminescent courtship-originated multiple times during evolution, allowing the empirical study of their longevity after careful phylogenetic and divergence time analyses. Here, we estimate the first transcriptome-based molecular phylogeny and divergence times of Cypridinidae. We report extreme longevity of bioluminescent courtship, a trait important in mate choice and probably under sexual selection. Our relaxed-clock estimates of divergence times coupled with stochastic character mapping show luminous courtship evolved only once in Cypridinidae-in a Sub-Tribe, we name Luxorina-at least 151 millions of years ago from cypridinid ancestors that used bioluminescence only in antipredator displays, defining a Tribe we name Luminini. This time-calibrated molecular phylogeny of cypridinids will serve as a foundation for integrative and comparative studies on the biochemistry, molecular evolution, courtship, diversification, and ecology of cypridinid bioluminescence. The persistence of luminous courtship for hundreds of millions of years suggests that sexual selection did not cause a rapid loss of associated traits, and that rates of speciation within the group exceeded extinction risk, which may contribute to the persistence of a diverse clade of signaling species. [Ancestral state reconstruction; Biodiversity; co-option; divergence time estimates; macroevolution; Ostracoda; phylogenomics; sexual selection.].


Identification of ADP in the iron-sulfur flavoprotein trimethylamine dehydrogenase.
Analysis of the 2.4-A resolution electron density map of trimethylamine dehydrogenase has revealed the unexpected presence of one molecule of ADP/subunit. This binding has been confirmed chemically. The binding site is located at the analogous position of the ADP moiety of FAD in glutathione reductase, the FAD and NADPH binding domains of which resemble two of the domains of trimethylamine dehydrogenase. Comparison of the environments of the ADP moieties in the two proteins indicates that 32 residues in 6 peptides are in equivalent positions with a root mean square deviation for C alpha positions of 1.11 A. Twelve of these amino acids are identical, based on the electron density-derived "x-ray" sequence of trimethylamine dehydrogenase. Detailed analysis of the environment of the ADP moiety indicates that most of the conserved residues are not in direct contact with the cofactor. Some of them probably represent the "fingerprint" of the beta alpha beta binding fold found in dinucleotide binding proteins, but the remaining conserved residues may indicate a closer evolutionary relationship between these two proteins.


The ecology of dippers Cinclus cinclus (L.) in relation to stream acidity in upland Wales: time-activity budgets and energy expenditure.
The time-activity budget and energy expenditure of a riverine bird, the dipper Cinclus cinclus, was studied from March 1988 to July 1989, across a range of streams of contrasting acidity in upland Wales. Differences in time-activity budgets of birds on acidic and circumneutral streams were consistent with documented differences in prey availability and diet. Birds spent a significantly greater proportion of their active day foraging, swimming and flying, and less time resting, on acidic streams. Activity measurements varied significantly through the year, through the day, and with river flow. Despite differences in time budgets, mean Daily Energy Expenditure (DEE) on acidic streams was only 4.5-7.0% greater than on circumneutral streams. However, rates of energy gain were greater for dippers on circumneutral streams for every month of the year, a pattern confirmed by differences in body condition. By spending more time feeding, dippers on acidic streams will have less time for other activities such as self-maintenance and predator surveillance; they may also be less able to meet the additional demands accompanying the initiation of breeding. These findings are discussed in relation to the feeding ecology and breeding performance of dippers on streams of contrasting water quality in upland Wales.


Pain of first-trimester abortion: a study of psychosocial and medical predictors.
Pain during first-trimester abortion by suction-curettage under local anesthesia alone was measured with the McGill Pain Questionnaire (MPQ), and verbal and visual analogue scales in 109 women. The average intensity of abortion pain ranked among moderately intense pain recorded with the MPQ. However, the pain scores had a wide range and appeared influenced by several demographic, psychosocial and medical variables. A correlation/stratified multiple regression design was used to examine the sources of individual variability in pain reports. The results indicated that over a third of the variance in pain reports could be predicted by a set of variables which included the patient's age, self-reports of pre-operative depression, anxiety, fear, low pain tolerance, social and moral concerns, and gynecological characteristics such as uterus retroversion, menstrual pain and gestational age. Pain scores were significantly higher for women who reported moderate to severe levels of pre-abortion depression and for younger patients (13-17 years); other psychosocial and gynecological features made small additional contributions to the prediction of pain scores. The implications of the results are discussed in relation to pain management strategies and underscore the special needs of each woman facing an abortion.


Outcome of periapical lesions in a rat model of type 2 diabetes: refractoriness to systemic antioxidant therapy.
This study evaluated the development of periapical lesions in a rat model of type 2 diabetes and assessed the potential actions of the antioxidant agent tempol in this model. Male Wistar rats were used; they received tap water (N = 5) or a 20% glucose solution (N = 15) during a period of 9 weeks. At the sixth week, periapical lesions were induced on the first mandibular molars, and the animals were subdivided into 4 groups. The subgroup 1 was composed of nondiabetic rats orally receiving saline solution (10 mL/kg). Chronically glucose-fed rats were divided into the following subgroups: (2) saline-treated animals (10 mL/kg by oral route), and animals treated with tempol by gavage at doses of (3) 50 mg/kg or (4) 100 mg/kg. The body weight was monitored thoroughly. After 21 days of apical periodontitis induction, the animals were killed, and the mandibles were collected and submitted to radiographic and histologic analysis. The livers were collected to determine free radicals, and the blood plasma was used to measure insulin levels. Type 2 diabetic rats displayed a significant decrease of body weight gain and a slight increase of insulin levels, which were allied to reduced levels of the antioxidant components catalase and reduced glutathione; these alterations were reversed by tempol. Concerning the periapical lesions, neither radiographic nor histologic analysis revealed any significant difference between control and type 2 diabetic rats. In diabetic rats, the apical periodontitis was refractory to tempol treatment. The extent and cellularity of periapical lesions in glucose-fed type 2 diabetic rats were similar to those seen in control rats. Despite affecting other parameters related to diabetes, tempol failed to improve the outcome of endodontic lesions in type 2 diabetic animals.


Comparative genomics of Paraburkholderia kururiensis and its potential in bioremediation, biofertilization, and biocontrol of plant pathogens.
Burkholderia harbors versatile Gram-negative species and is β-Proteobacteria. Recently, it was proposed to split the genus in two main branches: one of animal and plant pathogens and another, Paraburkholderia, harboring environmental and plant-beneficial species. Currently, Paraburkholderia comprises more than 70 species with ability to occupy very diverse environmental niches. Herein, we sequenced and analyzed the genome of Paraburkholderia kururiensis type strain KP23T , and compared to P. kururiensis M130, isolated in Brazil, and P. kururiensis susbp. thiooxydans, from Korea. This study focused on the gene content of the three genomes with special emphasis on their potential of plant-association, biocontrol, and bioremediation. The comparative analyses revealed several genes related to plant benefits, including biosynthesis of IAA, ACC deaminase, multiple efflux pumps, dioxygenases, and degradation of aromatic compounds. Importantly, a range of genes for protein secretion systems (type III, IV, V, and VI) were characterized, potentially involved in P. kururiensis well documented ability to establish endophytic association with plants. These findings shed light onto bacteria-plant interaction mechanisms at molecular level, adding novel information that supports their potential application in bioremediation, biofertilization, and biocontrol of plant pathogens. P. kururiensis emerges as a promising model to investigate adaptation mechanisms in different ecological niches.


Identification of the novel HLA-B*51:367 allele in a cord blood donor by next-generation sequencing.
The new HLA-B*51:367 differs from B*51:01:01:64 by four substitutions in exon 1.


[Electrophysiologic study of the connections of the 2d somatosensory area of the cortex with the neo-, paleocortical and striate formations of the brain].
2nd somatosensory area (C2) was shown to have modulating influence on evoked activity of the 1st somatosensory and visual areas. Electrical stimulation of C2 evoked short latency potentials in the structures of paleocortex (hippocampus) and striatum (putamen, caudate nucleus). Analysis of the data obtained here and elsewhere (19) suggests the important role of C2 in mechanisms of selection, comparison and integration of adequate sensory information under conditions of uncertainty, for the programming of behaviour.


Retrospective intra-scan motion correction.
This paper analyzes the effects of intra-scan motion and demonstrates the possibility of correcting them directly in k-space with a new automatic retrospective method. The method is presented for series of 2D acquisitions with Cartesian sampling. Using a reference k-space acquisition (corrected for translations) within the series, intra-scan motion parameters are accurately estimated for each trajectory in k-space of each data set in the series resulting in pseudo-random sample positions. The images are reconstructed with a Bayesian estimator that can handle sparse arbitrary sampling in k-space and reduces intra-scan rotation artefacts to the noise level. The method has been assessed by means of a Monte Carlo study on axial brain images for different signal-to-noise ratios. The accuracy of motion estimates is better than 0.1 degrees for rotation, and 0.1 and 0.05 pixel, respectively, for translation along the read and phase directions for signal-to-noise ratios higher than 6 of the signals on each trajectory. An example of reconstruction from experimental data corrupted by head motion is also given.


Temperature as a dependent variable in the study of cholinergic mechanisms.
1. Change in core temperature over time can be used as a dependent variable when studying the effects of manipulations on neurotransmitter systems. This article focuses on the measurement of core temperature as a strategy for detecting changes in the status of cholinergic systems. 2. Cholinergic neurons participate in the process of thermoregulation and interventions affecting them alter the thermal response to cholinomimetics. For example, chronic treatment with amitriptyline, chronic swim stress and inescapable footshock supersensitize rats to the hypothermic effects of oxotremorine. 3. This is consistent with the hypothesis that the pathophysiologies of tricyclic antidepressant withdrawal phenomena and stress involve supersensitivity of muscarinic mechanisms. 4. Uses of thermoregulation paradigms for investigating the actions of lithium ion, electroconvulsive shock and substances of abuse on muscarinic mechanisms are discussed. Applications to problems in the arena of clinical research are highlighted.


Histamine release into tracheal lumen and bronchial reactivity. The histamine liberation of proteolytic enzymes in the tracheal lumen in relation to the bronchomotoric reactivity.
The influence of tracheal lavage in a restricted area, with a proteolytic enzyme (Pronase) on the bronchial response to challenge with ACH and AE was studied in 15 dogs. Histamine concentrations measured in the tracheal liquid after lavage with Pronase were smaller than the values after tracheal lavage with allergen. Airway response to ACH inhalation was significantly increased after tracheal lavage with Pronase. Effects of proteolytic enzymes and allergens on the tracheal wall are compared. Allergens and proteolytic enzymes evidence a different mode of action in increasing reactivity of peripheral airways.


Cerebellar mutations affecting the postnatal survival of Purkinje cells in the mouse disclose a longitudinal pattern of differentially sensitive cells.
The pattern of surviving Purkinje cells (PCs) was investigated in three cerebellar mutant mice with severe postnatal PC death. Two of these mutations, nervous (nr) and Purkinje cell degeneration (pcd) mutations are already well characterized. The third mutation is a new one, which appeared spontaneously in DW/J-Pas mice and was called tambaleante (tbl). PCs were identified by immunocytochemistry using an antibody against vitamin D-dependent calcium-binding protein which labels all the PCs in adult control mice. In each of the three mutations, surviving PCs are arranged according to a different and reproducible pattern which is symmetric relative to the midline. In NR and young PCD mutants, PCs are closely packed in broad sagittal bands. In TBL, they are more loosely arranged in a rather patchy pattern. In PCD and in TBL mutants the death of resistant PCs is only shortly delayed but in NR there is little change in the number of surviving PCs after 3 months. The differential sensitivity of subsets of PCs to the effect of nr, pcd, and tbl mutations is topographically determined. These results provide a new evidence of the PC heterogeneity which has been previously demonstrated by histochemical and immunohistochemical techniques. Moreover, in the anterior vermis of control mice, three thin sagittal bands of PCs are labeled by the Q113 monoclonal antibody. Similarly, in the anterior lobe of the NR cerebellum, the thin longitudinal strips of missing PCs coincide with the absence of Q113 immunoreactivity: in this region the nr mutation affects specifically the survival of Q113 positive cells. However, other clusters of Q113 immunoreactive PCs do survive in NR mice suggesting that susceptibility to the nr mutation and Q113 positivity are two independent markers of the underlying PC compartmentalization.


Insulin-sensitive association of GLUT-4 with endocytic clathrin-coated vesicles revealed with the use of brefeldin A.
The interaction of the adipocyte/skeletal muscle glucose transporter (GLUT-4) with clathrin lattices may be important in maintaining its intracellular distribution. To better understand the role of clathrin lattices in the sorting of GLUT-4, we have attempted to determine the cellular origin of clathrin-coated vesicles (CCVs) that contain this transporter. The fungal toxin brefeldin A (BFA) causes the selective disassembly of clathrin lattices at the trans-Golgi network (TGN), but not at the plasma membrane (PM), thus providing a way of estimating the proportion of GLUT-4 in PM- versus TGN-derived clathrin lattices. Exposure of 3T3-L1 adipocytes to BFA resulted in a rapid disassembly of clathrin lattices at the TGN, observed by optical sectioning microscopy, and to a pronounced decrease in the yield of CCVs purified from these cells. Thus, CCVs isolated from BFA-treated cells are likely to be derived from the PM. Immunoblotting experiments revealed the presence of GLUT-4 in such CCVs, suggesting that under basal conditions the transporter is continually retrieved from the PM through the CCV pathway. Exposure of both BFA-treated or non-treated cells to insulin resulted in a 4-6-fold increase in the concentration of GLUT-4 at the PM. In parallel, the concentration of GLUT-4 in PM-derived CCVs decreased by 60%. These results suggest (a) that the effect of insulin to increase the cell surface concentration of GLUT-4 is not inhibited by BFA, and (b) that a decreased association of GLUT-4 with endocytic CCVs may be important in facilitating its increased cell surface concentration in response to the hormone.


Marital differences in blood pressure and the risk of hypertension among Polish men.
The aim of the study was to evaluate the relationship between marital status and blood pressure, and to assess the risk of hypertension in adult Polish men, after adjustment for BMI. Material comprised the data of 2,271 healthy men, aged 25-60, occupationally active inhabitants of Wroclaw (south-western Poland). Arterial hypertension was diagnosed when systolic blood pressure (SBP) > or =140 mm Hg and/or diastolic blood pressure (DBP) > or = 90 mm Hg. The following categories of marital status and educational level were applied: never married vs. currently married, and well-educated vs. poorly educated, respectively. The data on lifestyle elements were obtained from questionnaires. Multi-factorial analyses of covariance (ANCOVA) were used to compare mean values of SBP and DBP in married vs. never married in subsequent age categories with BMI as a covariate. Independent effects of marital status, life-style variables and body mass index (BMI) on the risk of hypertension in men were analysed using the multifactorial models of logistic regression. In our analysis an interesting epidemiological phenomenon was observed. Never married men had on average higher SBP and DBP than married men. Never married had also a higher risk of hypertension when compared to married men, even when adjusted for different demographic, socio-economic, life-style variables, and even that never married men had lower BMI than married subjects. Marital differences in psychological status (prolonged stress and low social support), dietary intake (mainly sodium and potassium intake) and economic aspects of living alone are suggested as factors, which might explain at least partly the marital diversity in blood pressure and the risk of hypertension in men.


Levels of PCDDs and PCDFs in Korean river sediments and their detection by biomarkers.
Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) are a group of toxic halogenated aryl hydrocarbons inducing various physiological disorders against biological organisms. Here, we investigated their levels in sediment samples taken from 12 different rivers in Korea. The levels of PCDD/PCDFs in sediment samples were expressed as concentrations and international TEQ values. Among 17 PCDD/PCDFs selected as target compounds in this study, the 1,2,3,4,7,8-HxCDD and OCDD were found in all river sediments with significant variation in various congener profiles of PCDD/PCDFs in sediments. PCDD/PCDFs could be monitored by sensitive biomarkers using insect immune system. Out of 12 river sediment samples, the biomarkers reported four spots (up, middle, and down Singil sites and Ansan) as putative contamination areas. When comparing both chemical and biological monitoring results, two methods agreed three spots of Singil as contamination areas (above 10ppt levels) as well as six river sediment samples as relatively less-contaminated areas, but differed in the results in Ansan and Miho, probably due to relative non-specificity of biomarkers. Despite some disparity between bio- and chemical monitoring results, the biomarkers can be recommended as a device warning the contamination of dioxins in the environment because of a fast and inexpensive detection method.


MiR196a-5p in extracellular vesicles released from human nasopharyngeal carcinoma enhance the phagocytosis and secretion of microglia by targeting ROCK1.
The microenvironment of the brain has become increasingly recognized as an essential regulator in metastatic and primary brain tumors. Recent studies demonstrate that circulating tumor-derived exosomes are critical for the brain tumor microenvironment. Nasopharyngeal carcinoma (NPC), a malignant tumor of the head and neck, often invades the skull base but infrequently extends to brain parenchyma. Neurobiological communication between microglia and tumor-derived extracellular vesicles (EVs) has been extensively studied, but how NPC cells regulate the immune microenvironment in the brain remains unknown. Here, we report that NPC derived EVs lead to increased microglial phagocytosis and proliferation, and heightened levels of IL-6, IL-8, CXCL1 and TGF-β1. Analysis of microRNAs in EVs reveal that miR196a-5p is the major effector microRNA. Moreover, we demonstrate an enrichment of miR196a-5p in the plasmatic EVs of NPC patients. Further investigation demonstrated that miR196a-5p was transferred to microglia and regulated microglial structure and functions by downregulating the expression of ROCK1. Therefore, these data indicate that NPC-derived EVs are potent modulators of microglial functions in brain microenvironment. Regardless of brain colonization, EVs-mediated functional changes in microglia may be a universal phenomenon that results in the alteration of the tumor host's microenvironment in the brain.


Involvement of steroids and cytochromes P(450) species in the triggering of immune defenses.
In vertebrates the wide variety of cytochromes P(450) (P(450)) is a key for elimination of low molecular weight xenobiotics and for the production and metabolism of steroid hormones. In contrast, xenobiotics of large molecular weight are processed and eliminated after the immune response. The suppression of immune response by native P(450)-produced glucocorticoid (GC) hormones constitutes a first link between P(450) and immunity. In the last decade, mechanisms and molecules responsible for the triggering of immune response were investigated and results showed that many tissues and organs transform native 3beta-hydroxysteroids into 7-hydroxylated metabolites that trigger immunity. Present data suggest that 7-hydroxysteroids are native anti-GCs that block the GC-induced immunosuppression. Because specific P(450) are responsible for the production of 7-hydroxylated steroids resulting into increased immunity, a second link exists between P(450) and immunity. Taken together, these findings support the proposal that P(450) are keys to all of the known defense mechanisms of vertebrates against all xenobiotic forms.


Ocular P2 receptors and glaucoma.
Adenosine triphosphate (ATP), an energy source currency in cells, is released or leaked to the extracellular space under both physiological and pathological conditions. Extracellular ATP functions as an intercellular signaling molecule through activation of purinergic P2 receptors. Ocular tissue and cells release ATP in response to physiological stimuli such as intraocular pressure (IOP), and P2 receptor activation regulates IOP elevation or reduction. Dysregulated purinergic signaling may cause abnormally elevated IOP, which is one of the major risk factors for glaucoma. Glaucoma, a leading cause of blindness worldwide, is characterized by progressive degeneration of optic nerves and retinal ganglion cells (RGCs), which are essential retinal neurons that transduce visual information to the brain. An elevation in IOP may stress RGCs and increase the risk for glaucoma pathogenesis. In the aqueous humor of human patients with glaucoma, the ATP level is significantly elevated. Such excess amount of ATP may directly cause RGC death via a specific subtype of P2 receptors. Dysregulated purinergic signaling may also trigger inflammation, oxidative stress, and excitotoxicity via activating non-neuronal cell types such as glial cells. In this review, we discussed the physiological roles of extracellular nucleotides in the ocular tissue and their potential role in the pathogenesis of glaucoma. This article is part of the Special Issue on 'Purinergic Signaling: 50 years'.


l-Arabinose Inhibits Colitis by Modulating Gut Microbiota in Mice.
l-Arabinose is a monosaccharide extracted from plants or fibers, which is known to have a variety of functional properties. In this study, we aim to investigate whether l-arabinose could inhibit colitis by modulating gut microbiota. l-Arabinose was administered in mice daily in a dextran sodium sulfate (DSS)-induced colitis model. The histological analysis, disease index, and the expression of inflammatory genes were measured. 16S-rRNA sequence analysis was performed to investigate gut microbiota. Intriguingly, we found that l-arabinose could repress DSS-induced colitis and inhibit p38-/p65-dependent inflammation activation. Besides that, our data revealed that l-arabinose-modulated DSS-induced gut microbiota were disturbed. Additionally, the perturbed gut microbiota was responsible for the suppressive effects of l-arabinose on DSS-induced colitis treated with antibiotics. Lastly, Caco-2 cells were used to confirm the protective effects of l-arabinose in colitis or inflammatory bowel disease. As expected, the protein expression levels in Caco-2 cells of pro-inflammatory genes, which were treated with l-arabinose and incubated with or without tumor necrosis factor alpha. Our work suggested that l-arabinose exerts anti-inflammation effects in DSS-induced colitis. These beneficial effects have correlations with the composition, diversity, and abundance of the gut microbiota regulated by l-arabinose. l-Arabinose could be a remarkable candidate as a functional food or novel therapeutic strategy for intestinal health.


Soluble antigen derived from IV larva of Angiostrongylus cantonensis promotes chitinase-like protein 3 (Chil3) expression induced by interleukin-13.
Angiostrongyliasis caused by Angiostrongylus cantonensis (A. cantonensis) is an emerging food-borne parasitic disease, which refers basically to eosinophilic meningitis. Chitinase-like protein 3 (Chil3), a member of chitinase-like protein family which has chemotactic activity for eosinophils, is reported to be highly upregulated in brain of mouse infected with A. cantonensis. The mechanisms of high expression of Chil3 and the association between A. cantonensis and Chil3 are rarely reported. In order to understand the mechanism of high expression of Chil3 in A. cantonensis-infected mouse, we measured the level of Chil3 in RAW 264.7 and BV2 cell lines stimulated with soluble antigen of A. cantonensis by qPCR and ELISA. To explore the role of Chil3 in inflammation caused by A. cantonensis, we extracted and cultured brain mononuclear cells (BMNCs) and detected the eosinophil chemotactic activity of Chil3 using transwell assay and flow cytometer. Furthermore, we treated the infected mice by injection with rmChil3 and then counted the number of larvae in brains of infected mice and treated mice to examine the association between the worm and Chil3. Our results showed the soluble antigen from A. cantonensis could promote the Chil3 expression in macrophage and microglial cell lines induced by interleukin-13. In conclusion, we supposed that high expression of Chil3 enhanced by soluble antigens from A. cantonensis might be the reason of serious eosinophil infiltration in mouse brain after A. cantonensis infection.


Effect of synbiotics on growth performance, gut health, and immunity status in pre-ruminant buffalo calves.
Synbiotics are employed as feed additives in animal production as an alternate to antibiotics for sustaining the gut microbiota and providing protection against infections. Dairy calves require a healthy diet and management to ensure a better future for the herd of dairy animals. Therefore, the present study was carried out to investigate the effect of synbiotics formulation on growth performance, nutrient digestibility, fecal bacterial count, metabolites, immunoglobulins, blood parameters, antioxidant enzymes and immune response of pre-ruminant Murrah buffalo calves. Twenty-four apparently healthy calves (5 days old) were allotted into four groups of six calves each. Group I (control) calves were fed a basal diet of milk, calf starter and berseem with no supplements. Group II (SYN1) calves were fed with 3 g fructooligosaccharide (FOS) + Lactobacillus plantarum CRD-7 (150 ml). Group III (SYN2) calves were fed with 6 g FOS + L. plantarum CRD-7 (100 ml), whereas calves in group IV (SYN3) received 9 g FOS + L. plantarum CRD-7 (50 ml). The results showed that SYN2 had the highest (P < 0.05) crude protein digestibility and average daily gain compared to the control. Fecal counts of Lactobacilli and Bifidobacterium were also increased (P < 0.05) in supplemented groups as compared to control. Fecal ammonia, diarrhea incidence and fecal scores were reduced in treated groups while lactate, volatile fatty acids and antioxidant enzymes were improved compared to the control. Synbiotic supplementation also improved both cell-mediated and humoral immune responses in buffalo calves. These findings indicated that synbiotics formulation of 6 g FOS + L. plantarum CRD-7 in dairy calves improved digestibility, antioxidant enzymes, and immune status, as well as modulated the fecal microbiota and decreased diarrhea incidence. Therefore, synbiotics formulation can be recommended for commercial use in order to achieve sustainable animal production.


Incidence, features, and outcomes of cytomegalovirus DNAemia in unmanipulated haploidentical allogeneic hematopoietic stem cell transplantation with post-transplantation cyclophosphamide.
Conflicting data have been published as to the risk of cytomegalovirus (CMV) DNAemia and CMV disease in patients undergoing haploidentical hematopoietic stem cell transplantation (haplo-HSCT) with post-transplantation cyclophosphamide. We conducted a multicenter retrospective study including 118 patients subjected to unmanipulated haplo-HSCT to further clarify this issue. An historic cohort comprising 165 patients undergoing other transplant modalities (HLA-matched related, matched unrelated or mismatched) was built for comparison purposes. Plasma CMV DNA monitoring was performed using two highly sensitive real-time PCR assays. Overall, the cumulative incidence of CMV DNAemia, recurrent CMV DNAemia, and CMV DNAemia requiring preemptive antiviral therapy in patients undergoing haplo-HSCT was 63.9%, 34.9%, and 50.1%, respectively. These figures were rather comparable for other transplant modalities (P = .22, P = .13 and P = .72, respectively). A trend toward longer duration of episodes and shorter CMV DNA doubling times was observed in haplo-HSCT patients in comparison with other transplant modalities. Furthermore, median CMV DNA peak load was significantly higher in haplo-HSCTs (P = .008), yet overall mortality by day 180 and 365 was the same across comparison groups. There were five cases of CMV disease, and all occurred in haplo-HSCT patients. This latter observation is worrying and merits further investigation. The incidence of initial and recurrent episodes of CMV DNAemia either requiring or not antiviral therapy in unmanipulated haplo-HSCT was comparable to other transplant modalities in our cohort.


Adverse drug reactions causing hospitalization can be monitored from computerized medical records and thereby indicate the quality of drug utilization.
To use computerized medical records to study the frequency of adverse drug reactions (ADR) as a cause for acute admission to a university hospital. Computerized medical records in 168 consecutively acutely admitted cases to a short-term internal medicine ward at a university hospital were retrospectively evaluated to see if an ADR could have caused the admission. In 18 cases (11%), an ADR was judged to be the reason for acute admission to the hospital. Augmented pharmacological effects (type A reactions) accounted for 89% of the cases. An additional eight cases (5%) were intentional intoxications with prescription drugs. ADR-associated hospital admissions were age-related (average age 72 compared to 65 in patients admitted for other reasons) and also associated with poly-pharmacy (8.3 drugs compared to 5.2). The ADR was considered to arise from a drug-drug interaction in four cases (22%). Although all ADRs fulfilled the criteria for mandatory reporting, none of them were reported to the Swedish Adverse Drug Reactions Advisory Committee (SADRAC). The observed frequency of ADR-related acute hospital admissions in this pilot study is similar to earlier reported data from Sweden and abroad. Almost all of these reactions were known pharmacological (type A) reactions and many therefore theoretically preventable. In contrast to many prospective and costly studies, this retrospective analysis of computerized medical records offered a simple method that can be used in routine health care and also serve as an indicator of the quality of drug use.


Assessment at UK medical schools varies substantially in volume, type and intensity and correlates with postgraduate attainment.
In the United Kingdom (UK), medical schools are free to develop local systems and policies that govern student assessment and progression. Successful completion of an undergraduate medical degree results in the automatic award of a provisional licence to practice medicine by the General Medical Council (GMC). Such a licensing process relies heavily on the assumption that individual schools develop similarly rigorous assessment policies. Little work has evaluated variability of undergraduate medical assessment between medical schools. That absence is important in the light of the GMC's recent announcement of the introduction of the UKMLA (UK Medical Licensing Assessment) for all doctors who wish to practise in the UK. The present study aimed to quantify and compare the volume, type and intensity of summative assessment across medicine (A100) courses in the United Kingdom, and to assess whether intensity of assessment correlates with the postgraduate attainment of doctors from these schools. Locally knowledgeable students in each school were approached to take part in guided-questionnaire interviews via telephone or Skype(TM). Their understanding of assessment at their medical school was probed, and later validated with the assessment department of the respective medical school. We gathered data for 25 of 27 A100 programmes in the UK and compared volume, type and intensity of assessment between schools. We then correlated these data with the mean first-attempt score of graduates sitting MRCGP and MRCP(UK), as well as with UKFPO selection measures. The median written assessment volume across all schools was 2000 min (mean = 2027, SD = 586, LQ = 1500, UQ = 2500, range = 1000-3200) and 1400 marks (mean = 1555, SD = 463, LQ = 1200, UQ = 1800, range = 1100-2800). The median practical assessment volume was 400 min (mean = 472, SD = 207, LQ = 400, UQ = 600, range = 200-1000). The median intensity (minutes per mark ratio) of summative written assessment was 1.24 min per mark (mean = 1.28, SD = 0.30, LQ = 1.11, UQ = 1.37, range = 0.85-2.08). An exploratory analysis suggested a significant correlation of total assessment time with mean first-attempt score on both the knowledge and the clinical assessments of MRCGP and of MRCP(UK). There are substantial differences in the volume, format and intensity of undergraduate assessment between UK medical schools. These findings suggest a potential for differences in the reliability of detecting poorly performing students, or differences in identifying and stratifying academically equivalent students for ranking in the Foundation Programme Application System (FPAS). Furthermore, these differences appear to directly correlate with performance in postgraduate examinations. Taken together, our findings highlight highly variable local assessment procedures that warrant further investigation to establish their potential impact on students.


Amphetamine-induced locomotor behavior of mice is influenced by DSIP.
The delta sleep-inducing peptide (DSIP) has been shown to induce effects other than only delta sleep. One of these effects was the paradoxical thermoregulatory and locomotor response of rats to amphetamine after DSIP administration. In the present investigation we found similar effects of DSIP on the locomotor activity in mice. However, two different doses of DSIP (30 and 120 nmol/kg) and 3 doses of amphetamine (4, 10, and 15 mg/kg) produced a complex pattern of effects in mice tested at 22 degrees C. In general, DSIP-treated mice showed lower locomotor activity after amphetamine than controls, but under two conditions, both using 15 mg/kg amphetamine, DSIP produced higher scores; this occurred in the first two hours after amphetamine for the 30 nmol/kg DSIP group and in the third hour for mice given 120 nmol/kg DSIP. The results indicate that the effects of DSIP on locomotor behavior were dependent on the dosage of the peptide and the time of measurement as well as the level of amphetamine stimulation.


Retraction: Down-regulated LncR-MALAT1 suppressed cell proliferation and migration by inactivating autophagy in bladder cancer.
[This retracts the article DOI: 10.1039/C8RA04876B.].


Astrocyte-Neuronal Communication and Its Role in Stroke.
Astrocytes are the most abundant glial cells in the central nervous system. These cells are an important hub for intercellular communication. They participate in various pathophysiological processes, including synaptogenesis, metabolic transformation, scar production, and blood-brain barrier repair. The mechanisms and functional consequences of astrocyte-neuron signaling are more complex than previously thought. Stroke is a disease associated with neurons in which astrocytes also play an important role. Astrocytes respond to the alterations in the brain microenvironment after stroke, providing required substances to neurons. However, they can also have harmful effects. In this review, we have summarized the function of astrocytes, their association with neurons, and two paradigms of the inflammatory response, which suggest that targeting astrocytes may be an effective strategy for treating stroke.


Glutamine supplementation attenuates ethanol-induced disruption of apical junctional complexes in colonic epithelium and ameliorates gut barrier dysfunction and fatty liver in mice.
Previous in vitro studies showed that glutamine (Gln) prevents acetaldehyde-induced disruption of tight junctions and adherens junctions in Caco-2 cell monolayers and human colonic mucosa. In the present study, we evaluated the effect of Gln supplementation on ethanol-induced gut barrier dysfunction and liver injury in mice in vivo. Ethanol feeding caused a significant increase in inulin permeability in distal colon. Elevated permeability was associated with a redistribution of tight junction and adherens junction proteins and depletion of detergent-insoluble fractions of these proteins, suggesting that ethanol disrupts apical junctional complexes in colonic epithelium and increases paracellular permeability. Ethanol-induced increase in colonic mucosal permeability and disruption of junctional complexes were most severe in mice fed Gln-free diet. Gln supplementation attenuated ethanol-induced mucosal permeability and disruption of tight junctions and adherens junctions in a dose-dependent manner, indicating the potential role of Gln in nutritional intervention to alcoholic tissue injury. Gln supplementation dose-dependently elevated reduced-protein thiols in colon without affecting the level of oxidized-protein thiols. Ethanol feeding depleted reduced protein thiols and elevated oxidized protein thiols. Ethanol-induced protein thiol oxidation was most severe in mice fed with Gln-free diet and absent in mice fed with Gln-supplemented diet, suggesting that antioxidant effect is one of the likely mechanisms involved in Gln-mediated amelioration of ethanol-induced gut barrier dysfunction. Ethanol feeding elevated plasma transaminase and liver triglyceride, which was accompanied by histopathologic lesions in the liver; ethanol-induced liver damage was attenuated by Gln supplementation. These results indicate that Gln supplementation ameliorates alcohol-induced gut and liver injury.


Intravenous heparin dosing strategy in hospitalized patients with atrial dysrhythmias.
Patients with non-valvular atrial fibrillation (AF) have an elevated stroke risk that is 2-7 times greater than in those without AF. Intravenous unfractionated heparin (UFH) is commonly used for hospitalized patients with atrial fibrillation and atrial flutter (AFL) to prevent stroke. Dosing strategies exist for intravenous anticoagulation in patients with acute coronary syndromes and venous thromboembolic diseases, but there are no data to guide providers on a dosing strategy for intravenous anticoagulation in patients with AF/AFL. 996 hospitalized patients with AF/AFL on UFH were evaluated. Bolus dosing and initial infusion rates of UFH were recorded along with rates of stroke, thromboemobolic events, and bleeding events as defined by the International Society on Thrombosis and Haemostasis criteria. Among 226 patients included in the analysis, 76 bleeding events occurred. Using linear regression analysis, initial rates of heparin infusion ranging from 9.7 to 11.8 units/kilogram/hour (U/kg/h) resulted in activated partial thromboplastin times that were within therapeutic range. The median initial infusion rate in patients with bleeding was 13.3 U/kg/h, while in those without bleeding it was 11.4 U/kg/h; p = 0.012. An initial infusion rate >11.0 U/kg/h yielded an OR 1.95 (1.06-3.59); p = 0.03 for any bleeding event. Using IV heparin boluses neither increased the probability of attaining a therapeutic aPTT (56.1 vs 56.3 %; p = 0.99) nor did it significantly increase bleeding events in the study (35.7 vs 31.3 %; p = 0.48). The results suggest that higher initial rates of heparin are associated with increased bleeding risk. From this dataset, initial heparin infusion rates of 9.7-11.0 U/kg/h without a bolus can result in therapeutic levels of anticoagulation in hospitalized patients with AF/AFL without increasing the risk of bleeding.


Opioid prescribing and risk of drug-opioid interactions in older discharged patients with polypharmacy in Australia.
Background Opioids are commonly prescribed to managing chronic pain in older persons. However, these patients are often at risk of drug-opioid interactions due to polypharmacy. Objectives To identify the prevalence of opioid prescribing and drug-opioid interactions in poly-medicated older patients and factors associated with opioid prescribing. Setting Patients were included if they were admitted to the Royal Adelaide Hospital between September 2015 and August 2016, aged ≥ 75 years and took ≥ 5 medications at discharge. Methods After ethics approval, data of were retrospectively collected from case notes. The Charlson Comorbidity Index and Drug Burden Index were determined and opioids were classified as strong or weak. The association between opioid use and concurrent medications was computed using logistic regression and the results presented as odds ratios (OR) and 95% confidence intervals (95% CI), adjusted for age, sex, Charlson Comorbidity Index, number of prescribed medications and modified-Drug Burden Index. Main outcome measure Association between concurrent medications and opioid prescribing. Results 15,000 geriatric admissions were identified, of which 1192 were included. A total of 283 (23.7%) patients were prescribed opioids, with oxycodone accounting for 56% of these prescriptions. Opioid users were prescribed more medications (11.2 vs. 9.0, P < 0.001) and had higher Drug Burden Index (1.2 vs. 0.14, P < 0.001) compared to non-users. Opioid use was associated with concurrent prescription of antiepileptics (OR = 1.7, 95% CI 1.1-2.6), and negatively associated with Charlson Comorbidity Index (OR = 0.9, 95% CI 0.8-0.98) and concurrent use of antipsychotics (OR = 0.5, 95% CI 0.3-0.9) and beta blocking agents (OR = 0.4, 95% CI 0.3-0.6). Conclusions Strong opioids were prescribed more often than weak opioids and opioid users presented with characteristics and concurrent medications which increased the risk of opioid related adverse drug effects.


Metalloprotease from Bacillus thuringiensis.
Bacillus thuringiensis var. kurstaki was shown to produce an extracellular, metal chelator-sensitive protease during the early stages of sporulation. Protease production in nutrient broth was dependent upon supplementation with Mn2+ or Ca2. The addition of Ca24 was required for enzyme stabilization...


The effect of a series of fixatives on the AgNOR technique.
With increasing interest being shown in nucleolar organizer regions (NORs) in pathology, it was considered of great importance to evaluate the effect of some of the more commonly used and more specialized fixatives on the demonstration of these moieties. NORs can be demonstrated in paraffin sections by a silver technique (AgNOR method) which was developed from a method used by cytogeneticists for the demonstration of NORs in chromosome spreads. The degree of staining is dependent on the fixation regime employed and results may vary greatly from one fixative to another. The fixative schedules and post-treatments used in this study were based on standard sequences from the literature. We have shown that, in general, alcohol-based fixatives give optimal results, Carnoy's fluid being especially recommended. Mercurial and dichromate-containing fixatives were found to have highly detrimental effects on NOR staining. 'Routine' 10 per cent formol saline fixation gave adequate results whereas 10 per cent neutral buffered formalin gave optimal staining, similar to alcohol-based fixation.


Segmental responses of abdominal motoneurons in decerebrate cats.
As a first step towards elucidating the synaptic organization underlying segmental responses of abdominal muscles I recorded the responses of branches of the left cranial (L(1)L) and caudal (L(2)L) and right caudal (L(2)R) lumbar (iliohypogastric) nerves to electrical shocks of different intensities to the caudal branch of L(2)L in nine decerebrate paralyzed and ventilated cats. If such reflex responses subserve a respiratory function, then they should be bilaterally similar; if they do not, lateral asymmetry should be evident. At intensities activating only large diameter axons (i.e. spindle and tendon organ afferents), stimulation typically elicited in the rostral branch of L(2)L a brief (approximately 1.6 ms) short-latency (approximately 1.8 ms) excitation followed by a suppression of activity (approximately 8-26 ms). Responses increased in amplitude as stimulus intensity increased, the suppression of activity being interrupted by an excitation (latency approximately 5. 4 ms, duration approximately 3.6 ms) in four cats. L(1)L responses were similar. Contralateral responses in the same segment (L(2)R) in five cats consisted of a suppression of activity in four, a short-latency (approximately 3.3 ms) excitation being present in three; increases in stimulus intensity in two additional cats elicited these excitatory and inhibitory responses. I conclude: (1) the variable responses between cats reflect differences in nerve bundles and, therefore, target muscles, from which the recordings were made; and (2) because of the lateral asymmetry of responses, abdominal afferent activation elicited postural (rotational) rather than respiratory reflexes.


Objective sonographic measures for characterizing myofascial trigger points associated with cervical pain.
The purpose of this study was to determine whether the physical properties and vascular environment of active myofascial trigger points associated with acute spontaneous cervical pain, asymptomatic latent trigger points, and palpably normal muscle differ in terms of the trigger point area, pulsatility index, and resistivity index, as measured by sonoelastography and Doppler imaging. Sonoelastography was performed with an external 92-Hz vibration in the upper trapezius muscles in patients with acute cervical pain and at least 1 palpable trigger point (n = 44). The area of reduced vibration amplitude was measured as an estimate of the size of the stiff myofascial trigger points. Patients also underwent triplex Doppler imaging of the same region to analyze blood flow waveforms and calculate the pulsatility index of blood flow in vessels at or near the trigger points. On sonoelastography, active sites (spontaneously painful with palpable myofascial trigger points) had larger trigger points (mean ± SD, 0.57 ± 0.20 cm(2)) compared to latent sites (palpable trigger points painful on palpation; 0.36 ± 0.16 cm(2)) and palpably normal sites (0.17 ± 0.22 cm(2); P < .01). Analysis of receiver operating characteristic curves showed that area measurements could robustly distinguish between active, latent, and normal sites (areas under the curve, 0.9 for active versus latent, 0.8 for active versus normal, and 0.8 for latent versus normal, respectively). Doppler spectral waveform data showed that vessels near active sites had a significantly higher pulsatility index (median, 8.3) compared to normal sites (median, 3.0; P < .05). The results presented in this study show that myofascial trigger points may be classified by area using sonoelastography. Furthermore, monitoring the trigger point area and pulsatility index may be useful in evaluating the natural history of myofascial pain syndrome.


Management of midface maxillofacial trauma.
The management of midface trauma continues to challenge maxillofacial surgeons. The complex local anatomy and functional and cosmetic importance of the region make precise surgical correction and reconstruction essential to success.


Iron Related Biomarkers Predict Disease Severity in a Cohort of Portuguese Adult Patients during COVID-19 Acute Infection.
Large variability in COVID-19 clinical progression urges the need to find the most relevant biomarkers to predict patients' outcomes. We evaluated iron metabolism and immune response in 303 patients admitted to the main hospital of the northern region of Portugal with variable clinical pictures, from September to November 2020. One hundred and twenty-seven tested positive for SARS-CoV-2 and 176 tested negative. Iron-related laboratory parameters and cytokines were determined in blood samples collected soon after admission. Demographic data, comorbidities and clinical outcomes were recorded. Patients were assigned into five groups according to severity. Serum iron and transferrin levels at admission were lower in COVID-19-positive than in COVID-19-negative patients. The levels of interleukin (IL)-6 and monocyte chemoattractant protein 1 (MCP-1) were increased in COVID-19-positive patients. The lowest serum iron and transferrin levels at diagnosis were associated with the worst outcomes. Iron levels negatively correlated with IL-6 and higher levels of this cytokine were associated with a worse prognosis. Serum ferritin levels at diagnosis were higher in COVID-19-positive than in COVID-19-negative patients. Serum iron is the simplest laboratory test to be implemented as a predictor of disease progression in COVID-19-positive patients.


Biodistribution and in vivo efficacy of genetically modified human mesenchymal stem cells systemically transplanted into a mouse bone fracture model.
Human mesenchymal stem cells (hMSCs) have generated a great deal of interest in clinical application due to their ability to undergo multi-lineage differentiation. Recently, ex vivo genetic modification of hMSCs was attempted to increase their differentiation potential. The present study was conducted to evaluate the biodistribution and in vivo efficacy of genetically modified hMSCs. To accomplish this, Runx2, which is a key transcription factor associated with osteoblast differentiation, was transduced into hMSCs using lentiviral vectors expressing green fluorescent protein (GFP) or luciferase. Here, we developed an experimental fracture in mice femur to investigate the effects of Runx2-transduced hMSCs on bone healing and migration into injury site. We conducted bio-luminescence imaging (BLI) using luciferase-tagged vector and quantitative real-time PCR using GFP probe to investigate the biodistribution of Runx2-transduced hMSCs in the fracture model. The biodistribution of hMSC cells in the fractured femur was observed at 14 days post-transplantation upon both BLI imaging and real-time PCR. Moreover, the fractured mice transplanted with Runx2-transduced hMSCs showed superior bone healing when compared to mock-transduced hMSC and MRC5 fibroblasts which were used as control. These data suggested that transplanted genetically modified hMSCs systemically migrate to the fractured femur, where they contribute to bone formation in vivo.


Oral triptans and nausea: treatment considerations in migraine.
The objective of this paper is to review evidence showing that migraine patients who are nauseated before using oral triptans tend to have a poor treatment response, as well as to establish a framework for further investigation of the association between response to oral medications and pretreatment nausea among migraineurs. SYNTHESIS AND CONCLUSION: In patients with migraine, pretreatment nausea predicts a poor response to oral triptans. This finding may be inherent in the oral route of delivery of medication, as pretreatment nausea is associated with gastric stasis, which can impair absorption of oral medications and reduce therapeutic efficacy. In addition, oral triptans contribute to the development of nausea among migraine patients who are nausea free before they treat, perhaps because oral tablet use triggers or exacerbates nausea in the same manner as eating or drinking among patients who are nauseated or vulnerable to nausea. Importantly, these observations are derived from a small evidence base and post-hoc analyses or, in the case of treatment-emergent nausea, adverse event reports. Further assessment of the relationships between nausea and oral triptans is necessary before drawing firm conclusions. Should these observations be validated, the use of oral triptans in migraine attacks with nausea or in patients prone to nausea should be reevaluated. Novel routes of administration for triptans allow patients to receive the benefits of migraine-specific therapy even when oral therapy is suboptimal.


A cryogenic 14-channel 13 C receiver array for 3T human head imaging.
This article presents a novel 14-channel receive-only array for 13 C human head imaging at 3 T that explores the SNR gain by operating at cryogenic temperature cooled by liquid nitrogen. Cryostats are developed to evaluate single-coil bench SNR performance and cool the 14-channel array with liquid nitrogen while having enough thermal insulation between the coils and the sample. The temperature distribution for the coil array is measured. Circuits are adapted to the -189°C environment and implemented in the 14-channel array. 13 C images are acquired with the array at cryogenic and room temperature in a 3T scanner. Compared with room temperature, the array at cryogenic temperature provides 27%-168% SNR improvement over all voxels and 47% SNR improvement near the image center. The measurements show a decrease of the element noise correlation at cryogenic temperature. It is demonstrated that higher SNR can be achieved by cryogenically cooling the 14-channel array. A cryogenic array suitable for clinical imaging can be further developed on the array proposed. The cryogenic coil array is most likely suited for scenarios in which high SNR deep in a head and decent SNR on the periphery are required.


Magnetic nanoparticles based on cerium MOF supported on the MWCNT as a fluorescence quenching sensor for determination of 6-mercaptopurine.
In this study, a new magnetic nanocomposite was developed as an efficient and fast-response fluorescence quenching sensor for determination of anticancer drug 6-mercaptopurine (6-MP). For this purpose, the needle-shape fluorescence metal-organic framework of cerium (Ce-MOF) were successfully synthesized on the surface of multiwalled carbon nanotubes using 1,3,5-benzenetricarboxylic acid ligand via a facile solvothermal assisted route and magnetized. The accuracy of the proposed synthesis was confirmed using the FT-IR, FE-SEM, XRD, and VSM methods. The obtained product as presented the fluorescence emission in 331 nm by excitation of 293 nm in excitation/emission slit widths of 10.0 nm. The operation of suggested method is based on quenching the fluorescence signal in accordance with increasing the 6-MP concentration. The proposed assay effectively detected the trace amount of 6-MP in the linear range of 1.0 × 10-6 to 7 × 10-5 M. The limit of detection and limit of quantification were obtained as 8.6 × 10-7 and 2.86 × 10-6 M, respectively. The analyte molecule was determined in real samples with satisfactory recoveries between 98.75 and 105.33.


Optogenetics in primates: a shining future?
To understand the functional role of specific neurons in micro- and macro-brain circuitry, health, and disease, it is critical to control their activity precisely. This ambitious goal was first achieved by optogenetics, allowing researchers to increase or decrease neural activity artificially with high temporal and spatial precision. In contrast to the revolution optogenetics engendered in invertebrate and rodent research, only a few studies have reported optogenetic-induced neuronal and behavioral effects in primates. Such studies are nonetheless critical before optogenetics can be applied in a clinical setting. Here, we review the state-of-the-art tools for performing optogenetics in mammals, emphasizing recent neuronal and behavioral results obtained in nonhuman primates.


Isolation of embryonic stem (ES) cells in media supplemented with recombinant leukemia inhibitory factor (LIF).
The isolation of pluripotent murine embryonic stem (ES) cells has previously been achieved by coculturing the ES cells with fibroblast feeder cells. In this report we demonstrate that ES cell lines can be isolated from murine 129/Sv He blastocysts in the absence of feeder cells in culture medium supplemented with recombinant leukemia inhibitory factor (LIF). Three of the ES cell lines (MBL-1, MBL-2, and MBL-3) were isolated by directly explanting blastocysts, whilst two ES cell lines (MBL-4 and MBL-5) were isolated from blastocysts pretreated by immunosurgery. Three of the ES cell lines contained the Y chromosome (MBL-1, MBL-2, and MBL-5) with a high proportion of the cells displaying a normal diploid karyotype with a modal chromosome number of 40. All of the ES cell lines tested expressed the stem cell markers ECMA-7 and alkaline phosphatase, which were lost on removal of LIF when the ES cells differentiated into a variety of cell types. The full developmental potential of the ES cells was determined by injecting cells from two of the independently derived ES cell lines, MBL-1 and MBL-5, into C57BL/6J blastocysts. A high proportion of the pups born were chimeric as judged by coat pigmentation. Subsequent breeding established that the ES cells had contributed to the germ line. These results demonstrate that feeder cells are not essential for the isolation of pluripotent ES cell lines.


Acculturation Timing among Newcomer and more Experienced Immigrant Youth: The Role of Language Use in Ethnic Friendship Homophily.
The usage of the new language is a crucial aspect in immigrant youth adaptation. However, despite substantial inter- and intraindividual variability and dynamic changes, language usage has been studied primarily with a focus on static interindividual differences. This study utilized a recently introduced Temporal Model of Acculturative Change to test associations between language acquisition and friendship homophily. More specifically, three concepts were tested: pace (individual rate of change), relative timing (the deviation from peers with similar length of residence), and transition timing (preparedness for the relocation). Data comprised a three-wave-longitudinal sample of 820 ethnic German adolescents from Eastern European States who immigrated to Germany (Mage = 16.1, 57% girls). Results revealed, particularly among recent immigrant adolescents, that transition timing predicted earlier relative acculturation timing in language usage and that early relative timing in language usage predicted levels and change rates in friendship homophily (over and above acculturation pace and the actual level of language usage). Findings highlight the need to better understand the dynamics in acculturation processes of immigrant youth.


Necroptosis-related lncRNAs: establishment of a gene module and distinction between the cold and hot tumors in glioma.
Gliomas are the most common primary tumors of the central nervous system and portend a poor prognosis. The efficacy of emerging and promising immunotherapies varies significantly among individuals. Distinction and transformation of cold and hot tumors may improve the antitumor efficacy of immunotherapy. In this study, we constructed a necroptosis-related lncRNA module based on public databases. The association of this module with survival was assessed using the Cox regression, Kaplan-Meier survival analysis, and nomogram, external validation was also conducted in another public database. Furthermore, we performed gene set enrichment analysis (GSEA), immune checkpoint and tumor microenvironment analysis, and in vitro qRT-PCR validation. Finally, we clustered all samples into 2 clusters based on the expression of model lncRNAs and identified cluster 1 as cold tumors with fewer infiltrating T cells. Identifying cold and hot tumors by necroptosis-related lncRNAs can help available immunotherapeutic strategies to achieve efficacy in the precise treatment of individuals. Prior treatment failure can be overcome by targeting necroptosis-related lncRNAs.


The early diagnosis of acute myocardial infarction. Comparison of a simple algorithm with a computer program for electrocardiogram interpretation.
The sensitivity and specificity of electrocardiographic (ECG) interpretation by a simple algorithm was compared with a computer read ECG machine. Clinical data and ECG findings on 264 consecutive patients admitted to a coronary care unit with suspected acute myocardial infarction were prospectively entered into an algorithm with 13 end-points. These end-points were compared with the interpretations of a computer read ECG machine (Marquette MAC PC). 86 patients (32.5%) had confirmed acute infarction. 85% of those with infarction had some form of ST elevation on their initial ECG. Patients with ST elevation presented earlier (4.9 +/- 4.9 versus 8.0 +/- 9.7 hours after symptom onset, p < 0.001), and were older (66.5 +/- 11.0 versus 62.0 +/- 12.5 years, p < 0.01) than those without infarction. According to the algorithm 94.2% of patients with infarction had some form of ECG abnormality, compared with 55.6% of those without infarction (p < 0.001). The area under the receiver operating characteristic (ROC) curve of the algorithm was 92.3% of the area of the graph. This was more (p < 0.01) than the area under the ROC curve of the interpretations of the computer read ECG machine (83.9%). Marked ST elevation with reciprocal changes was the most specific indicators of infarction (Likelihood ratio 51.7). The algorithm, therefore, was comparatively sensitive and specific in the early diagnosis of acute infarction.


Determination of the stable conformation of GABA(A)-benzodiazepine receptor bivalent ligands by low temperature NMR and X-ray analysis.
The stable conformations of GABA(A)-benzodiazepine receptor bivalent ligands 2 and 3 were determined by low temperature NMR spectroscopy and confirmed by single crystal X-ray analysis. The linear conformation was important for these dimers to access the binding site and exhibit potent in vitro affinity as illustrated for alpha5 subtype selective ligand 2 (15 nM). Bivalent ligand 3 with the 5 atom linker folded back upon itself both in solution and in the solid state, moreover, it did not bind to Bz receptors.


Grazoprevir plus peginterferon and ribavirin in treatment-naive patients with hepatitis C virus genotype 1 infection: a randomized trial.
Grazoprevir (MK-5172, Merck & Co., Inc.) is a selective inhibitor of the hepatitis C virus (HCV) NS3/4a protease. The aim of this study was to evaluate the safety and efficacy of grazoprevir at doses of 25-100 mg/day in combination with peginterferon and ribavirin (PEG-IFN/RBV). In this randomized, dose-ranging, multicentre trial, treatment-naive adults with chronic HCV genotype 1 infection received once-daily grazoprevir 25 mg, 50 mg or 100 mg plus PEG-IFN/RBV for 12 weeks. Patients with quantifiable HCV RNA (≥25 IU/mL) at week 4 received an additional 12 weeks of PEG-IFN/RBV. The primary endpoint was sustained virologic response (HCV RNA <25 IU/mL 12 weeks after completing therapy [SVR12]). Eighty-seven patients were randomly assigned and received ≥1 dose of therapy. Median time to undetectable HCV RNA was 16 days in the 100-mg arm and 22 days in the 25- and 50-mg arms. All patients except one had HCV RNA undetectable or unquantifiable at week 4 and received 12 weeks of therapy. SVR12 was achieved by 13 of 24 (54.2%), 21 of 25 (84.0%) and 23 of 26 (88.5%) patients in the 25-, 50- and 100-mg arms, respectively (per-protocol analysis). Three patients discontinued as a result of nonserious adverse events (AEs) and three patients experienced serious AEs. Transaminase elevations occurred in two patients (one each in the 25- and 100-mg arms). These data support further study of the grazoprevir 100-mg dose. Phase 3 studies of grazoprevir 100 mg in combination with elbasvir are currently ongoing (NCT01710501; protocol P038).


Periodic nocturnal myoclonus in a patient with hyperexplexia (startle disease).
The periodic nocturnal myoclonus of a patient with hyperexplexia has been studied. Evidence has been given that the jerks are spontaneous arousal reactions. The temporal characteristics of the jerks have been analysed. The jerks appear to be correlated with the respiratory rhythm and the data suggest a correlation of the jerks with circulatory and respiratory higher order waves. The results are discussed with relation to the literature concerning spontaneous sleep jerks.


High-quality superconducting α-Ta film sputtered on the heated silicon substrate.
Intrigued by the discovery of the long lifetime in the α-Ta/Al2O3-based Transmon qubit, researchers recently found α-Ta film is a promising platform for fabricating multi-qubits with long coherence time. To meet the requirements for integrating superconducting quantum circuits, the ideal method is to grow α-Ta film on a silicon substrate compatible with industrial manufacturing. Here we report the α-Ta film sputter-grown on Si (100) with a low-loss superconducting TiNx buffer layer. The α-Ta film with a large growth temperature window has a good crystalline character. The superconducting critical transition temperature (Tc) and residual resistivity ratio (RRR) in the α-Ta film grown at 500 °C are higher than that in the α-Ta film grown at room temperature (RT). These results provide crucial experimental clues toward understanding the connection between the superconductivity and the materials' properties in the α-Ta film and open a new route for producing a high-quality α-Ta film on silicon substrate for future industrial superconducting quantum computers.