Promiscuity comes at a price: catalytic versatility vs efficiency in different metal ion derivatives of the potential bioremediator GpdQ.
The glycerophosphodiesterase from Enterobacter aerogenes (GpdQ) is a highly promiscuous dinuclear metallohydrolase with respect to both substrate specificity and metal ion composition. While this promiscuity may adversely affect the enzyme's catalytic efficiency its ability to hydrolyse some organophosphates (OPs) and by-products of OP degradation have turned GpdQ into a promising candidate for bioremedial applications. Here, we investigated both metal ion binding and the effect of the metal ion composition on catalysis. The prevalent in vivo metal ion composition for GpdQ is proposed to be of the type Fe(II)Zn(II), a reflection of natural abundance rather than catalytic optimisation. The Fe(II) appears to have lower binding affinity than other divalent metal ions, and the catalytic efficiency of this mixed metal center is considerably smaller than that of Mn(II), Co(II) or Cd(II)-containing derivatives of GpdQ. Interestingly, metal ion replacements do not only affect catalytic efficiency but also the optimal pH range for the reaction, suggesting that different metal ion combinations may employ different mechanistic strategies. These metal ion-triggered modulations are likely to be mediated via an extensive hydrogen bond network that links the two metal ion binding sites via residues in the substrate binding pocket. The observed functional diversity may be the cause for the modest catalytic efficiency of wild-type GpdQ but may also be essential to enable the enzyme to evolve rapidly to alter substrate specificity and enhance k(cat) values, as has recently been demonstrated in a directed evolution experiment. This article is part of a Special Issue entitled: Chemistry and mechanism of phosphatases, diesterases and triesterases.


A hypothalamic endorphinic lesion attenuates acquisition of cocaine self-administration in the rat.
The present study explores the role of beta-endorphin-producing neurons of the arcuate nucleus in the behavioral effects of cocaine (i.e. acquisition of cocaine self-administration). Eight-week-old female rats were treated with a single estradiol valerate injection that causes a progressive lesion that is specific to beta-endorphin-producing neurons throughout the arcuate nucleus. Cocaine acquisition was suppressed following estradiol valerate pretreatment, while water reinforced behavior was similar to controls. Since estradiol valerate treated rats exhibit low estrogen plasma levels, estrogen replacement was performed but cocaine self-administration acquisition remained suppressed. In addition, analysis of beta-endorphin, dopamine, and DOPAC tissue levels confirmed the specificity of the endorphinic lesion resulting from estradiol valerate treatment. The suppression of cocaine self-administration acquisition following estradiol valerate treatment provides evidence for a significant role for beta-endorphin in cocaine reward.


Epilepsy and driving in North Carolina: an exploratory study.
Epileptic drivers offer a risk to the general driving population, both because of affected brain function and because of possible effects of medication. A 1982 pilot study examined the driving records of 112 persons using North Carolina Division of Health Services clinics for the treatment of epilepsy who also held a North Carolina driver's license. Of those undergoing treatment in the clinics, 26% were known by the Division of Motor Vehicles (DMV) to be epileptics. This group had a reported crash rate 1.4 times that of the general driving population, whereas the rate for epileptics not known to the DMV was 1.1 times the general rate. Epileptics with grand mal and temporal or psychomotor seizures accounted for all recorded crashes. Implications for highway safety administrators and for future research are discussed.


Primary structure of the heterosaccharide of the surface glycoprotein of Methanothermus fervidus.
The outer surface of the cells of the hyperthermophile Methanothermus fervidus is covered by crystalline glycoprotein subunits (S-layer). From the purified S-layer glycoprotein, a heterosaccharide was isolated. The heterosaccharide consists of D-3-O-methylmannose, D-mannose, and D-N-acetylgalactosamine in a molar ratio of 2:3:1 corresponding to a relative molecular mass of 1061.83 Da. 3-O-methylmannose could be partly replaced by 3-O-methylglucose. The primary structure of the glycan was revealed by methylation analysis, by plasma desorption mass spectrometry, and by high field NMR spectroscopy. The purified heterosaccharide is linked via N-acetylgalactosamine to an asparagine residue of the peptide moiety. The following structure is proposed for the heterosaccharide: alpha-D-3-O-MetManp-(1-->6)-alpha-D-3-O-MetManp-((1-->2)-alp ha-D-Manp)3-(1-->4) - D-GalNAc.


Butenyl-spinosyns, a natural example of genetic engineering of antibiotic biosynthetic genes.
Spinosyns, a novel class of insect active macrolides produced by Saccharopolyspora spinosa, are used for insect control in a number of commercial crops. Recently, a new class of spinosyns was discovered from S. pogona NRRL 30141. The butenyl-spinosyns, also called pogonins, are very similar to spinosyns, differing in the length of the side chain at C-21 and in the variety of novel minor factors. The butenyl-spinosyn biosynthetic genes (bus) were cloned on four cosmids covering a contiguous 110-kb region of the NRRL 30141 chromosome. Their function in butenyl-spinosyn biosynthesis was confirmed by a loss-of-function deletion, and subsequent complementation by cloned genes. The coding sequences of the butenyl-spinosyn biosynthetic genes and the spinosyn biosynthetic genes from S. spinosa were highly conserved. In particular, the PKS-coding genes from S. spinosa and S. pogona have 91-94% nucleic acid identity, with one notable exception. The butenyl-spinosyn gene sequence codes for one additional PKS module, which is responsible for the additional two carbons in the C-21 tail. The DNA sequence of spinosyn genes in this region suggested that the S. spinosa spnA gene could have been the result of an in-frame deletion of the S. pogona busA gene. Therefore, the butenyl-spinosyn genes represent the putative parental gene structure that was naturally engineered by deletion to create the spinosyn genes.


Determination of high mobility group A1 (HMGA1) expression in hepatocellular carcinoma: a potential prognostic marker.
Our objective was to investigate the expression of HMGA1 mRNA and protein in hepatocellular carcinoma (HCC) and the correlation between its expression and clinical pathological characteristics and prognosis. HMGA1 expression was determined at both the mRNA level and the protein level in 30 HCC tissues and their corresponding paracancer liver tissues (PCLTs) and 2 normal liver tissues by RT-PCR and IHC. Follow-up study was done on the 30 patients involved in this research. HMGA1 mRNA was detected in nine cases of HCC tissues and two PCLTs, for a positivity rate of 30% and 6.7%, respectively (P < 0.05), whereas no HMGA1 mRNA expression was found in normal liver tissues. Clinicopathological analysis revealed that HMGA1 mRNA expression was significantly correlated with Edmondson's grade (P < 0.05). HMGA1 protein was detected in four HCC tissues by IHC and located mainly in the nuclei; no positive staining was found in PCLTs. Follow-up study showed that HMGA1 mRNA-positive patients had a higher risk of recurrence/metastasis and a shorter survival than negative cases (P < 0.05). Our findings indicate that HMGA1 may be involved in the carcinogenesis and invasiveness of HCC and the determination of HMGA1 can be of great value in predicting the prognosis of patients with HCC.


E. coli interactions, adhesion and transport in alumino-silica clays.
Bacterial adhesion and transport in the geological formation are controlled by their mutual complex interactions, which have been quantified by the traditional and extended Derjaguin-Landau-Verwey-Overbeek (DLVO) theory as well as direct atomic force microscopy (AFM) measurements. In this research, the DLVO forces calculated based on the independently determined bacterial and porous media surface thermodynamic properties were compared with those of AFM measurements. Although differences in the order of several magnitudes existed, forces obtained from both ways could explain the observations of E. coli attachment to alumino-silica clays evaluated in laboratory columns under saturated and steady-state flow conditions. E. coli deposition in alumino-silica clays was simulated using a two-site convection-dispersion transport model against E. coli transport breakthrough curves, which was then linked to the interactions forces. By exploring the differences of the two force measurements, it was concluded that the thermodynamic calculations could complement the direct force measurements in describing bacterial interactions with the surrounding environment and the subsequent transport in the porous media.


No wrong decisions in an all-wrong situation. A qualitative study on the lived experiences of families of children with diffuse intrinsic pontine glioma.
Diffuse intrinsic pontine glioma (DIPG) is a rare, but lethal pediatric brain tumor with a median survival of less than 1 year. Existing treatment may prolong life and control symptoms, but may cause toxicity and side effects. In order to improve child- and family-centered care, we aimed to better understand the treatment decision-making experiences of parents, as studies on this topic are currently lacking. The data for this study came from 24 semistructured interviews with parents whose children were diagnosed with DIPG in two children's hospitals in Switzerland and died between 2000 and 2016. Analysis of the dataset was done using reflexive thematic analysis. For most parents, the decision for or against treatment was relatively straightforward given the fatality of the tumor and the absence of treatment protocols. Most of them had no regrets about their decision for or against treatment. The most distressing factor for them was observing their child's gradual loss of independence and informing them about the inescapability of death. To counter this powerlessness, many parents opted for complementary or alternative medicine in order to "do something." Many parents reported psychological problems in the aftermath of their child's death and coping strategies between mothers and fathers often differed. The challenges of DIPG are unique and explain why parental and shared decision-making is different in DIPG compared to other cancer diagnoses. Considering that treatment decisions shape parents' grief trajectory, clinicians should reassure parents by framing treatment decisions in terms of family's deeply held values and goals.


Bicuculline-induced enhancement of sensory responses and cross-correlations between reticular formation and cortical neurons.
The visual, auditory and somatosensory responses of neurons in the brain stem reticular formation (RF) and pericruciate cortex of the cat are enhanced by intravenous administration of subconvulsant doses of bicuculline. The degree of enhancement in RF neurons is somewhat greater in magnitude and occurs in a greater percentage of RF neurons. The latency of response is shorter in the RF than in the cortex in 70% of cases. A large percentage of simultaneously recorded RF and cortical neurons which became responsive to the same stimulus exhibited consistent convulsant-induced cross-correlations of firing which were not present before drug treatment. The latency and correlation data are consistent with the possibility that the RF may subserve the cortical enhancement. Auditory response thresholds in RF neurons are reduced by bicuculline administration. Enhancement of RF neuronal responsiveness has previously been observed with several other convulsant drugs which are thought to act on different neurotransmitters suggesting that it may reflect a general action of these agents beyond the effects on specific neurotransmitters. The bicuculline-induced correlation of firing of RF and pericruciate neurons may be involved in the mechanism of initiation of convulsant-mediated seizure generalization induced by sensory stimuli.


Interactions of trichloroethylene with DNA in vitro and with RNA and DNA of various mouse tissues in vivo.
The covalent binding of 14C-1,1,2-trichloroethylene (14C-TRI) metabolites to calf thymus DNA in vitro and to RNA and DNA of mouse brain, lung, liver, kidney, spleen, pancreas, and testis after repeated i.p. injections has been studied. Hydrolysates of DNA reacted with 14C-TRI in vitro and hydrolysates of RNA and DNA from selected organs were separated on Aminex A6 for quantitation of alkylation products. The presence of 3,N4-etheno(deoxy)cytidine, 1,N6-etheno(deoxy)adenosine and 1,N6-ethenoadenine was investigated. No radioactivity could be registered in DNA incubated with 14C-TRI in the absence of liver microsomes. Covalent binding of 14C-TRI to DNA took place in the presence of liver microsomes from control mice. The binding was enhanced by 50% if liver microsomes from phenobarbital pretreated mice were used. The radioactivity in DNA reacted with 14C-TRI and microsomes from control mice was eluted in early fractions and together with thymidine. The same two peaks appeared on chromatography of DNA incubated with 14C-TRI and liver microsomes from phenobarbital pretreated mice. In addition, radioactivity was eluted together with 1,N6-ethenoadenine. Radioactivity was registered in RNA and DNA from all of the studied organs after i.p. injections of 14C-TRI. The radioactivity in RNA increased in the order brain less than testis less than pancreas less than kidney less than liver less than lung less than spleen. The radioactivity in DNA increased in the order brain less than kidney less than testis less than lung less than pancreas less than liver less than spleen. Aminex A6 chromatography revealed that the entire radioactivity in RNA from liver and kidney and in DNA from kidney, testis, lung, pancreas, and spleen was due to metabolic incorporation, particularly into guanine and adenine. This finding indicates that the C-C bond in TRI is split, with the formation of C1-fragments, during biotransformation in vivo. In liver DNA, the metabolic incorporation of radioactivity was insignificant. Instead, the dominant part of the radioactivity in liver DNA was eluted in early fractions. The elution profile of radioactivity in liver DNA gave no direct evidence of the formation of TRI-DNA adducts in vivo. No etheno-derivatives were identified as alkylation products of TRI in vivo, which is consistent with current theories of the metabolic fate of TRI.


Sterol regulatory element binding protein 1 interacts with pregnane X receptor and constitutive androstane receptor and represses their target genes.
Sterol regulatory element binding protein 1 (SREBP-1) is a lipogenic transcription factor of the basic helix-loop-helix family. SREBP-1 binds to sterol regulatory elements (SREs) in the promoter of lipogenic genes and induces fatty acid and triglyceride synthesis. Decreased drug clearance has been observed in obese and other dyslipidemic rodents as well as in diabetic, obese or overfed rodents. A hallmark of these conditions is increased expression of SREBP-1 in the liver. We therefore searched for a possible link between regulation of cytochromes P450 (CYPs) and SREBP-1. We combined gene expression analysis, lipid analysis, effects of high levels of SREBP-1 in hepatocyte cultures to characterize the effects and protein interaction and chromatin immunoprecipitation assays to define the underlying mechanism. Finally, mice were fed a diet enriched in cholesterol to demonstrate the relevance of our data in vivo. By analyzing gene expression and lipids in cholesterol-fed mice or transfection of recombinant SREBP-1 in hepatocyte cultures the effect on CYPs was characterized. By use of protein interaction assays and chromatin immunoprecipitation the underlying mechanism was defined. We observed that SREBP-1 represses drug-mediated induction of hepatic CYPs, mainly members of the 2B and the 3A subfamilies. These drugs induce transcription of CYPs and other drug metabolizing enzymes via activation of the nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR). Here we report that the activation of SREBP-1 by insulin or cholesterol in mouse liver and primary human hepatocytes inhibits the transcriptional effects in PXR and CAR. Our results suggest that SREBP-1 functions as a non-DNA binding inhibitor and blocks the interaction of PXR and CAR with cofactors such as steroid receptor coactivator 1. Consequently, mRNA induction of CYPs by drugs and other xenochemicals is impaired. We conclude that PXR and CAR respond to lipid accumulation by direct interaction with SREBP-1 and show that drug metabolism and lipid metabolism are interconnected within a complex network of transcriptional regulators.


Celebrating scientific excellence and global health impact: The 2022 Canada Gairdner Awards.
From mRNA vaccines to community interventions in global child health, the 2022 Canada Gairdner awards demonstrate the importance of fundamental science and its translation into improved human health and well-being.


[Myxovirus structural investigations by spin probe methods. II. Influence of rimantadine on the structure of viral and artificial lipid membranes].
The lipid membrane structure of two myxoviruses (influenza and Sendai viruses), red blood cells and liposomes (prepared from total lipids of Sendai virus) have been investigated in the presence of rimantadine by means of two spin probes C5 and C6--the iminoxyl derivatives of stearic acids. Rimantadine was shown to penetrate the lipid membranes of myxoviruses and red blood sells and change the structure of the lipid phase no less than 0.8 nm apart from the viral surface. In the depth of 2.2 nm the structure of these lipid membranes remains unchanged after addition of rimantadine. Rimantadine also displaces the phase transition points of cell and viral lipid membranes to lower temperatures. Rimantadine does not change the lipid bilayer structure of liposomes 0.8 nm apart from its surface. These finding allow to suggest that rimantadine affects the cell and viral lipoprotein membranes.


Synthesis of novel 6'-spirocyclopropyl-5'-norcarbocyclic adenosine phosphonic Acid analogues as potent anti-hiv agents.
Novel 5'-norcarbocyclic adenosine phosphonic acid analogues with 6'-electropositive moiety such as spirocyclopropane were designed and synthesized from the commercially available diethylmalonate 5. Regioselective Mitsunobu reaction proceeded in the presence of an allylic functional group at a low reaction temperature in polar cosolvent [dimethylformamide (DMF)/1,4-dioxane] to give purine analogue 15. To improve cellular permeability and enhance the anti-human immunodeficiency virus (HIV) activity of this phosphonic acid, a SATE phosphonodiester nucleoside prodrug 23 was prepared. The synthesized adenosine phosphonic acids analogues 18-21 and 23 were subjected to antiviral screening against HIV-1.


Application of a non-species dependent ELISA for the detection of antibodies in sera of Burkholderia pseudomallei-immunized goats.
A modified, non-species dependent ELISA was performed to detect antibodies in sera of Burkholderia pseudomallei-immunized goats using protein G- or protein A-peroxidase conjugates. The rise of antibody titers during the immunization period exhibited corresponding results by modified ELISA comparison to conventional ELISA and the IHA. Regarding the increase of antibody levels from the pre-immunized baseline to the post-immunized status, the antibody titer detected by modified ELISA was higher than IHA but lower than conventional ELISA. Further efforts are needed to improve the modified ELISA for the diagnosis of melioidosis in animals.


The preventive effect of NR2B and NR2D-containing NMDAR antagonists on Aβ-induced LTP disruption in the dentate gyrus of rats.
Amyloid β-protein (Aβ) in the brain of Alzheimer's disease (AD) potently inhibits the synaptic plasticity subsequently causing the cognitive deficits. Long-term potentiation (LTP) of synaptic transmission is thought to be an important cellular mechanism underlying memory formation. Different NR2 subunits are involved in NMDA receptor-dependent LTP. In the present study, we investigated the roles of NR2B and NR2D-containing NMDAR on Aβ(1-42)-induced LTP deficits in the hippocampal slices of rats by using selective NMDAR antagonists. First, we found that Aβ(1-42) significantly inhibited the LTP in the dentate gyrus of slices as reported before. Following that the Aβ(1-42)-induced LTP inhibition was prevented by the pre-perfusion of the specific NR2B-containing NMDAR antagonists ifenprodil (approximately >200-fold selectivity for NR2B) and Ro25-6981 (>3,000-fold selectivity for NR2B), as well as PPDA, a specific NR2D receptor antagonist. Meanwhile, the antagonists on their own had no or only partial effects on the normal LTP in the same dose condition. These findings not only support the effects of NR2B and NR2D subunits on Aβ(1-42)-induced LTP deficits, but also imply that preferentially targeting NR2B- and NR2D-containing NMDARs may provide an effective means to prevent cognitive deficits in the early AD.


Analysis of NF1 transcriptional regulatory elements.
Mutations of the NF1 locus cause neurofibromatosis 1 (NF1), a clinically variable autosomal dominant disease. Expression of neurofibromin, the protein product of the NF1 gene, is regulated in a tissue- and developmentally-specific fashion, but the basis for this regulation is not understood. We used phylogenetic footprinting and other bioinformatic methods to identify potential transcriptional regulatory regions in the 5' upstream region and intron 1 of the NF1 gene from human, mouse, rat, and pufferfish. Three genomic segments that have equal or higher homology than the coding region were found in the NF1 5' upstream region, and four more very highly homologous regions were found in intron 1. Five of these highly homologous regions are confidently predicted to contain transcription factor binding sites. One highly homologous segment in the 5' upstream region spans the transcription start site and contains several potential transcription factor binding sites. This segment includes a novel 24-bp sequence (acttccggtggggtgtcatggcgg) that lies 310-333 bp upstream of the translation initiation site. This sequence, which is identical in human, mouse, and rat and differs by only 1-bp in Fugu, may contain the core promoter element for NF1 transcription.


Comparison of cyclosporin A, verapamil, PSC-833 and cremophor EL as enhancing agents of VP-16 in murine lymphoid leukemias.
Although verapamil, cyclosporin A. cremophor EL and PSC-833 are active as multidrug resistance modulators, there has been limited study of these compounds as possible chemotherapy enhancing agents against drug-sensitive tumors. We compared these agents as modifiers of VP-16 cytotoxicity in vitro and modifiers of VP-16 efficacy in vivo against drug-sensitive P388 and L1210 leukemias. Our study indicates that cyclosporin A enhances VP-16 cytotoxicity to a significantly greater extent than equimolar concentrations of verapamil or PSC-833. Although cremophor EL shows significantly greater activity than verapamil in VP-16 cytotoxicity enhancement in vitro, it is ineffective when added to VP-16 therapy of mice bearing L1210 leukemia.


Age-related differences in vitamin D status in Polish centenarians compared with 65-year-olds.
Vitamin D status is known to change with age. However, little is known about vitamin D status in centenarians. The aim of the study was to assess vitamin D status and correlations among the levels of parathyroid hormone (PTH), 25‑hydroxycholecalciferol (25[OH]D), 1,25-dihydroxycholecalciferol (1,25[OH]D), calcium, inorganic phosphorus, and alkaline phosphatase (ALP) activity in centenarians. The study group included 97 participants: 81 women and 16 men (median [interquartile range [IQR]) age, 101.4 [100.5-102.16] years). Centenarians were visited at their homes where examinations were conducted and blood samples collected. The control group consisted of 57 elderly subjects: 35 women and 22 men (median [IQR] age, 65.9 [65.3-66.5] years). The concentrations of PTH, 25(OH)D, and 1,25(OH)D were measured in frozen plasma samples, and calcium, phosphorus, and ALP levels, in serum samples. The median calcium level was 8.88 mg/dl in centenarians versus 9.52 mg/dl in 65‑year-old subjects (P <0.01); ALP, 223 IU versus 190 IU (P = 0.01); phosphorus, 3.01 mg/dl versus 3.23 mg/dl (P = 0.13); PTH, 45.59 pg/ml versus 29.27 pg/ml (P <0.01); 25(OH)D, 7.39 ng/ml versus 19.81 ng/ml (P <0.01); 1,25(OH)D, 57.5 pmol/l versus 78.6 pmol/l (P <0.01). Only centenarians demonstrated correlations among the measured laboratory parameters. Considering lower 25(OH)D, 1,25(OH)D, and calcium concentrations in the majority of centenarians, as well as the negative correlation between vitamin D active metabolites and PTH, vitamin D and calcium should be systematically supplemented in the oldest of the elderly.


Characterization of a hypermutable strain of Drosophila simulans.
A hypermutable strain of Drosophila simulans that originated from a single spontaneous mutant male was characterized. Seven different mutations were isolated from roughly 100 generations of offspring. The genetic analysis of the viable mutants showed two mutations on the X chromosome, one in the lozenge locus and the other in the ruby gene. The autosomic mutations characterized were a dpp-heldout-like, a blistered-like and a homoeotic dominant mutant with an antenna-to-leg transformation and ectopic eyes that we called Zoinho-napata.


[Epidemiological and clinical characteristics of dissociative disorders and somatoform disorders in Burkina Faso].
The aim of this study was to determine the epidemiological and clinical features of dissociative and somatoform disorders in the psychiatry department of the Centre Hospitalier Universitaire Yalgado Ouédraogo, in Burkina Faso. We carried out a retrospective and descriptive study over five years, from January 2003 to December 2007, of patients admitted in the psychiatric department for dissociative and somatoform disorders. The data were extracted from the medical notes and registers of consultation. After identification of the patients, the medical notes were exploited with a structured collect form for data on sociodemographic characteristics, life events and ICD-10 diagnoses. All the data were recorded by the same person. Data analysis was performed with Epi info 2007. During a period of 5 years, 3967 patients were registered, 179 had the diagnosis of dissociative and somatoform disorders (3.65% of the admissions) and 145 were included in our study. The majority of the patients were followed as outpatients (71%). Women were more represented with a sex ratio of four women to one man. The majority of the patients were young with an average age of 30 and mostly single (57.9%). The majority of the patients had a low academic standard, in connection with young age of the population. A striking biographic event was found in the majority of the cases; essentially family difficulties and death. Neurological symptoms were the most frequent in 63% of the cases. According to ICD-1O, dissociative disorders were most frequent (65.6%). Depression was mainly associated in15 cases. Dissociative disorders and somatoform disorders are frequent in our hospital; we must overcome the difficulties due to the complexity of the diagnosis and the cultural resistances to modern health care to determine the real prevalence of these disorders among the population in Burkina Faso.


Unusual cause of cerebral calcifications in an 8-year-old girl.
Genetic counseling and genetic screening for hyperoxaluria should be recommended for children with urinary lithiasis for early management to avoid progression to oxalosis especially if there is a family history of lithiasis. Primary hyperoxaluria type 1 (PH1) is caused by a deficiency of the liver peroxisomal enzyme alanine-glyoxylate aminotransferase (AGT) resulting in overproduction of calcium oxalates. In its later stage, a systemic deposit of calcium oxalates is observed. We present the case of an 8-year-old girl with exceptional neurological involvement secondary to this disease.


Time to reperfusion and other procedural characteristics of emergency coronary artery bypass surgery after unsuccessful coronary angioplasty.
A databank search was performed and 148 consecutive patients (mean age 59.5 +/- 10.4 years) were identified who underwent emergency coronary artery bypass surgery at the Mayo Clinic between November 20, 1979, and February 12, 1992, immediately after unsuccessful coronary angioplasty. At the end of the angioplasty procedure, there was no anterograde coronary blood flow in the treated artery in 54%, ongoing chest pain in 78%, and hemodynamic compromise requiring intravenous vasopressor therapy in 25% of patients; 127 patients (86%) had at least 1 of these adverse characteristics. After leaving the catheterization laboratory, the median time to arrival in the operating room was 12 minutes. Median time from arrival in the operating room to initiation of cardiopulmonary bypass was 86 minutes, to administration of cardioplegia was 98 minutes, and to removal of the aortic cross-clamp was 135 minutes. In-hospital mortality was 11%, and 18% developed nonfatal Q-wave myocardial infarction. Thus, significant time is required to achieve surgical reperfusion after unsuccessful coronary angioplasty.


The binding of isocolchicine to tubulin. Mechanisms of ligand association with tubulin.
Isocolchicine is a structurally related isomer of colchicine altered in the methoxytropone C ring. In spite of virtual structural homology of colchicine and isocolchicine, isocolchicine is commonly believed to be inactive in binding to tubulin and inhibiting microtubule assembly. We have found that isocolchicine does indeed bind to the colchicine site on tubulin, as demonstrated by its ability to competitively inhibit [3H]colchicine binding to tubulin with a KI approximately 400 microM. Isocolchicine inhibits tubulin assembly into microtubules with an I50 of about 1 mM, but the affinity of isocolchicine for the colchicine receptor site, 5.5 +/- 0.9 x 10(3) M-1 at 23 degrees C, is much less (approximately 500-fold) than that of colchicine. Unlike colchicine, isocolchicine binds rapidly, and the absorption and fluorescence properties of the complex are only modestly altered compared to free ligand. It is proposed that the binding of isocolchicine to tubulin may be rationalized either in terms of conformational states of colchicinoids when liganded to tubulin or by the structural requirements for C-10 substituents for high affinity binding to the colchicine receptor.


Evaluation of different measures of functional connectivity using a neural mass model.
We use a neural mass model to address some important issues in characterising functional integration among remote cortical areas using magnetoencephalography or electroencephalography (MEG or EEG). In a previous paper [Neuroimage (in press)], we showed how the coupling among cortical areas can modulate the MEG or EEG spectrum and synchronise oscillatory dynamics. In this work, we exploit the model further by evaluating different measures of statistical dependencies (i.e., functional connectivity) among MEG or EEG signals that are mediated by neuronal coupling. We have examined linear and nonlinear methods, including phase synchronisation. Our results show that each method can detect coupling but with different sensitivity profiles that depended on (i) the frequency specificity of the interaction (broad vs. narrow band) and (ii) the nature of the coupling (linear vs. nonlinear). Our analyses suggest that methods based on the concept of generalised synchronisation are the most sensitive when interactions encompass different frequencies (broadband analyses). In the context of narrow-band analyses, mutual information was found to be the most sensitive way to disclose frequency-specific couplings. Measures based on generalised synchronisation and phase synchronisation are the most sensitive to nonlinear coupling. These different sensitivity profiles mean that the choice of coupling measures can have dramatic effects on the cortical networks identified. We illustrate this using a single-subject MEG study of binocular rivalry and highlight the greater recovery of statistical dependencies among cortical areas in the beta band when mutual information is used.


Effects of experience and available cues on estrous versus diestrous preferences in male prairie voles, Microtus ochrogaster.
Male prairie vole preferences for estrous versus diestrous females and associated stimuli were investigated. The role of sexual experience in engendering preferences proved more complex than reported for other species. Naive males did not display preferences. Neither males receiving sexual experience through monogamous cohabitation, nor males housed with two females displayed preferences. Males exposed to both estrous and diestrous females, and males housed with other males and females in a semi-naturalistic setting, displayed preferences. Thus, preferences seem to be "tuned" by experiential manipulations. Cues available for discrimination also influenced the display of preferences, as different results were obtained using apparatus that differed in cue availability. The observed pattern of responsiveness to sex odors may be related to the social organization and mating system of the prairie vole, a mammal that is believed to be monogamous.


Change in Youth Mental Health During the COVID-19 Pandemic in a Majority Hispanic/Latinx US Sample.
Children and adolescents, individuals from racial and ethnic minority groups, and those with mental health conditions may be at greater risk for worsened mental health because of the COVID-19 pandemic. This study examined change in mental health from before to during the pandemic among predominantly Hispanic/Latinx adolescents. A total of 322 young adolescents (mean age = 11.99 years, 55% female and 45% male), with a racial/ethnic composition of 72.7% Hispanic/Latinx, 9.3% Black or African American, 5.9% multiple races, 5.0% Asian, 1.6% White, and 1.2% American Indian, completed a mental health screening measure prior to the COVID-19 pandemic and at 3 time-points beginning 1 month after COVID-19 stay-at-home measures were implemented. A subsample also completed a survey about their experience at home during COVID-19. Repeated-measures mixed analysis of covariance was used to evaluate change in each mental health domain, and whether youths who had elevated symptoms at baseline differed in their level of change, controlling for age and gender. For youths who had elevated levels of mental health problems before the pandemic, symptoms were significantly reduced across domains during the pandemic. Reductions in internalizing, externalizing, and total problems were clinically significant. For other youths, there were statistically significant reductions in internalizing and total problems, and no change in attention or externalizing problems. Post hoc analyses revealed that better family functioning was consistently related to lower mental health symptoms in youths during COVID-19 follow-ups. COVID-19 stay-at-home regulations may offer protective effects for youth mental health. Study results may be specific to this population of predominantly Hispanic/Latinx youths from a large city in the southwestern United States.


Long-acting somatostatin analog controls acid and gastrin secretion in benign, not in malignant, Zollinger-Ellison syndrome.
The long-acting somatostatin (SMS) analog, SMS 201-995 has beneficial effects on APUDomas. In two Zollinger-Ellison syndrome (ZES) patients we assessed basal acid output (BAO) and 24-h pH under SMS and compared them to controls. We also assessed total gastrin, gastrin 17, insulin, glucagon, C-peptide, and SMS by radioimmunoassay. In the benign gastrinoma, an acid-controlling action of SMS was shown, elevating the 24-h pH threshold over the pH range 1.5-5 of 55-10% compared with control. A parallel inhibition of the gastrins greater than 90% was apparent. We found no beneficial effect on gastric acid secretion and on tumor gastrin in the malignant gastrinoma despite a fourfold higher plasma SMS level. Non-tumor-related peptides were suppressed by approximately 50% and in contrast to gastrin they again reached pre-SMS levels before the next dose of the drug. We conclude that SMS is more effective in benign than in malignant gastrinomas, and may be exclusively so.


Reassessing the role of antitachycardia pacing in fast ventricular arrhythmias in primary prevention implantable cardioverter-defibrillator recipients: Results from MADIT-RIT.
In Multicenter Automatic Defibrillator Implantation Trial - Reduce Inappropriate Therapy (MADIT-RIT), high-rate cutoff (arm B) and delayed therapy (arm C) reduced the risk of inappropriate implantable cardioverter-defibrillator (ICD) interventions when compared with conventional programming (arm A); however, appropriate but unnecessary therapies were not evaluated. The purpose of this study was to assess the value of antitachycardia pacing (ATP) for fast ventricular arrhythmias (VAs) ≥ 200 beats/min in patients with primary prevention ICD. We compared ATP only, ATP and shock, and shock only rates in patients in MADIT-RIT treated for VAs ≥ 200 beats/min. The only difference between these randomized groups was the time delay between ventricular tachycardia detection and therapy (3.4 seconds vs 4.9 seconds vs 14.4 seconds). In arm A, 11.5% patients had events, the initial therapy was ATP in 10.5% and shock in 1%, and the final therapy was ATP in 8% and shock in 3.5%. In arm B, 6.6% had events, 4.2% were initially treated with ATP and 2.4% with shock, and the final therapy was ATP in 2.8% and shock in 3.8%. In arm C, 4.7% had events, 2.5% were initially treated with ATP and 2.3% with shock, and the final therapy was ATP in 1.4% and shock in 3.3%. The final shock rate was similar in arm A vs arm B (3.5% vs 3.8%; P = .800) and in arm A vs arm C (3.5% vs 3.3%; P = .855) despite the marked discrepancy in initial ATP therapy utilization. In MADIT-RIT, there was a significant reduction in ATP interventions with therapy delays due to spontaneous termination, with no difference in shock therapies, suggesting that earlier interventions for VAs ≥ 200 beats/min are likely unnecessary, leading to an overestimation of the value of ATP in primary prevention ICD recipients.


Effects of exogenous and endogenous histamine on the respiratory system.
There is a statistically strong correlation between the increase of the histamine concentration in the arterial plasma and the increase of airway resistance. The histamine causes a reflex bronchoconstriction on the sensory receptors. Histamine in low concentration increases the unspecific reaction in the bronchial system. This can also be achieved using low concentrations of allergen aerosols. Patients with chronic obstructive bronchitis show high concentrations of histamine in the sputum. The bronchoconstrictive potency of this histamine was found to be not always active. Higher concentrations of histamine can be liberated by washing the inner surface of the bronchial tree. Histamine injected in similar concentrations as that measured in the sputum causes bronchoconstriction. There should be high concentrations of histamine receptors as well as mast cells near the surface of the mucosa of the bronchial wall. There are many questions remaining open, which will be discussed.


Metabolic profiling and pharmacokinetic studies of sinapine thiocyanate by UHPLC-Q/TOF-MS and UHPLC-MS/MS.
Sinapine thiocyanate (ST) is an index component and pharmacological active component of Semen Sinapis and Semen Raphani, and it is widely used to relieving cough and asthma. This study aimed to obtain the metabolic and pharmacokinetic characterization of ST. The metabolic profiles of ST were obtained from rat plasma, urine, and feces via ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UHPLC-Q/TOF-MS). Thirteen metabolites were structurally identified, and the proposed metabolic pathways of ST included deamination, demethylation, hydrogenation, dehydration, and extensive conjugation, including glucuronidation and sulfonation. ST was selected as the plasma marker for the pharmacokinetic study. A simple and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for the quantitation of ST in rat plasma. The linear range of ST was 0.1-500 ng/mL (R2 = 0.9976), and the lowest limit of quantification was 0.1 ng/mL. The intra-precision and inter-precision of the assay were 1.31-5.12% and 2.72-7.66%, and the accuracy (RE%) ranged from - 4.88% to 3.82% and - 3.47% to 6.18%. The extraction recovery, matrix effect, and stability of ST were within acceptable limits. The established method was validated and successfully applied to the pharmacokinetic study of ST. For pharmacokinetic experiments, the male Sprague-Dawley rats were administrated with ST solution intravenously (2 mg/kg) or orally (100 mg/kg). The oral absolute bioavailability of ST was calculated as 1.84%, and the apparent volume of distribution of intravenous and intragastric administrations were 107.51 ± 21.16 L/kg and 78.60 ± 14.44 L/kg, respectively. The maximum plasma concentration was 47.82 ± 18.77 nM, and the time to maximum peak was 88.74 ± 20.08 min for the intragastric dosing group. According to the pharmacokinetic and metabolic profiling results, metabolites with high abundance of ST in bio-fluids would be the next object in tissue distribution and pharmacodynamic study.


Unique chemical activity in porous YbB2C2 ceramics with high porosity and high compressive strength.
High purity layered YbB2C2 powder is synthesized by a boro/carbothermic reduction method using YbBO3, B4C and graphite powders as raw materials. Its X-ray diffraction data are presented, and the space group P4/mbm (No. 127) is confirmed. The lattice parameters are a = b = 5.3389 Å and c = 3.5683 Å, and the atom positions are Yb (0.0000, 0.0000, 0.0000), B (0.3621, 0.8621, 0.5000), and C (0.1606, 0.6606, 0.5000). Porous YbB2C2 ceramics have a high porosity in the range of 69.89-58.11% and a high compressive strength in the range of 19.49-63.44 MPa. Furthermore, the as-produced porous YbB2C2 ceramics show unique chemical activity. Porous YbB2C2 ceramic with a porosity of 69.89% emits so much heat that it can burn a piece of paper when this ceramic is wetted by water. The rate of reaction between the porous YbB2C2 ceramic and water can be simply controlled by adjusting the porosity. The solid reaction products are YbB6, C and an unknown amorphous phase.


Right renal artery arising from the celiac artery - A rare anatomical variation.
Variations in origin and number of renal artery are common. We present a very rare anatomical variation where superior right renal artery was arising from celiac artery. This variation has important surgical implication.


ISAAC-based asthma and atopic symptoms among Ha Noi school children.
Childhood asthma and atopy prevalence patterns in the developing world are only beginning to be defined. No such information exists for Vietnam. Estimates would assist in anticipating health service needs as well as add to the growing database on global patterns of atopy. To estimate the prevalence of atopic symptoms in school children in Ha Noi, Vietnam, a cross-sectional survey was conducted of children aged 5- to 11-years-old in two schools using the parent self-administered International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire. The response rate was 66.4% (969 responses). The overall prevalence of selected symptoms were: 'ever wheezed' 24.9%, 'wheezed in past 12 months' 14.9%, 'ever had asthma' 12.1%, 'doctor-diagnosed asthma' 13.9%, 'ever experienced allergic rhinitis (AR) symptoms' 34.9%, 'AR-conjunctivitis symptoms in past 12 months' 10.7%, 'ever had hay fever' 7.8%, 'doctor-diagnosed hay fever' 11.2%, 'ever had eczema' 3.3% and 'doctor-diagnosed eczema' 3.2%. Kappa statistics demonstrated high within symptom category consistency for 'ever had asthma/doctor-diagnosed asthma' (0.728) and 'ever had eczema/doctor-diagnosed eczema' (0.906). Age and gender adjusted odds ratios (OR) were also consistently significant across wheeze and allergic rhinitis symptom categories [highest OR = 10.10 (95% CI 6.23-16.35) between allergic rhinoconjunctivitis and wheeze in past 12 months]. There is a high prevalence of ISAAC-based symptoms in school children in Ha Noi, Vietnam, often above global averages. The high level of association between atopic symptoms suggests some degree of reliability and validity. Childhood atopy symptom prevalence in Vietnam is more similar to that in developed countries rather than developing countries.


Genetic and environmental determinants of musical ability in twins.
Analyses of musical ability data from the Loehlin and Nichols National Merit Scholarship study are presented. Musical ability is indexed by four measures: interest in a profession in music, performance in school, performance outside of school, and receiving honors in music. These variables pose a challenge for behavior genetic analysis since they do not conform to the assumptions of traditional linear models. For example, there is a dependent relationship between the honors and the performance variables; one cannot obtain honors without performance. Several methods were employed to deal with these relationships, and the following conclusions appeared regardless of the method used. First, twin correlations were always high, ranging from 0.44 to 0.90 in monozygotic (MZ) twins and from 0.34 to 0.83 in dizygotic (DZ) twins. Second, although there was evidence for heritable variation, the effects of common environment were almost always larger than the effects of heredity. Third, marital assortment was not of sufficient magnitude to account for these common environment effects. In the young adults in this sample, musical ability is influenced more by shared family environment than by shared genes.


Two pharmacologically distinct histamine receptors mediating membrane hyperpolarization on identified neurons of Aplysia californica.
Two distinct hyperpolarizing responses are produced when histamine is iontophoretically applied onto the somal membranes of identified neurons within the cerebral ganglion of Aplysia: a biphasic response consisting of a rapid component (less than 5 sec) usually superimposed upon a slowly developing component; or a monophasic slowly developing response 5-20 sec in duration. The reversal potential values for the fast (typically -65 mV) and the slow (typically -89 mV) responses, and their shift to new values when the external potassium or chloride concentrations were altered, revealed that the fast and slow potentials are produced predominantly by conductance increases to chloride and potassium ions, respectively. The effects of histamine H1- and H2-receptor agonists and antagonists were studied to characterize the pharmacological properties of histamine receptors mediating these two ionically dissimilar hyperpolarizing responses. The slow potassium-dependent hyperpolarization could be mimicked by several histamine analogues; the most potent tested were the H1-receptor agonist, 2-methylhistamine, and the H2-receptor agonist, 4-methylhistamine. Neither of these agents mimicked the fast chloride-dependent histamine response. The slow potassium-dependent responses induced by histamine or histamine agonists were completely and reversibly blocked by the H2-receptor antagonist, cimetidine. By contrast, the slow potassium-dependent hyperpolarizations produced by iontophoretically applied acetylcholine or by dopamine to the same neurons were unaffected by cimetidine. Other H1 and H2 antagonists tested were either ineffective, or only partially blocked the slow hyperpolarizations in a non-selective manner. The fast chloride-dependent hyperpolarizations were not selectively antagonized by any of the H1 or H2 reagents tested, although they were effectively suppressed by tubocurarine and strychnine. These data indicate that two pharmacologically distinct histamine receptors mediate potassium- and chloride-dependent hyperpolarizations in Aplysia neurons. Neither of these receptors, however, could be classified as strictly H1 or H2 according to criteria presently used in non-neuronal tissues. The selectivity and reversibility of cimetidine indicate that this particular antihistaminic could be a valuable pharmacological tool for defining putative histaminergic synapses in Aplysia and perhaps other nervous systems.


Using mean field approach independent component analysis to fatty acid characterization with overlapped GC-MS signals.
In this paper, mean field independent component analysis (MF-ICA) was applied as a deconvolution method to separate complex gas chromatographic-mass spectrometric (GC-MS) signals obtained from fatty acid analysis of fish oil. The separation which is a blind operation was used as a complementary method in identification of the unknown components of a mixture and in quantification purposes, as well. In MF-ICA, the sources (mass spectra) are recovered from the mean of their posterior distributions and mixing matrix (chromatograms) and noise level are estimated through the maximum a posterior (MAP) solution. The number of independent components (ICs) in the overlapping signals can be estimated by the difference between the reconstructed and original GC-MS data. It was found that the chromatographic profiles and the mass spectra of the components in overlapping multicomponent GC-MS data can be accurately recovered with and without previously background correction. The resolved mass spectral sources satisfactory are identified using mass spectral search system. The recovered chromatographic area and the relative content of each analyte considering selected number of ICs are calculated and the results are compared with the ones obtained previously by using heuristic evolving latent projections (HELP) method.


Inflammatory conditions promote a switch of oligosaccharyltransferase (OST) catalytic subunit isoform expression.
Oligosaccharyltransferase (OST) complex catalyzes the N-glycosylation of nascent polypeptides in the endoplasmic reticulum. Glycoproteins are critical for normal cell-cell interactions, especially during an immune response. Abnormal glycosylation is an insignia of several inflammatory diseases. However, the mechanisms that regulate the differential N-glycosylation are not fully understood. The OST complex can be assembled with one out of two catalytic subunits, STT3A or STT3B, which have different enzymatic properties. In this work, we investigated the expression of STT3A and STT3B in several mouse models such as a crossbreeding of normal and abortion-prone mice and an intestinal inflammation model. These animals were either exposed or not to acoustic stress (acute or chronic). The expression of the isoforms was analysed by immunohistochemistry and protein immunoblot. Finally, we investigated the gene regulatory elements employing public databases. Results demonstrated that inflammation alters the balance between the expression of both isoforms in the affected tissues. In homoeostatic conditions, STT3A expression predominates over STT3B, especially in epithelial cells. This relation is reversed as a consequence of inflammation. An increase in STT3B activity was associated to the generation of mannose-rich N-glycans. Accordingly, this type of N-glycans were found to decorate diverse inflamed tissues. The STT3A and STT3B genes are differentially regulated, which could account for the differences in the expression levels observed here. Our results support the idea that these isoforms could play different roles in cellular physiology. This study opens the possibility of studying the STT3A/STT3B expression ratio as a biomarker in acute inflammation or chronic diseases.


Does puerperal illness distinguish a subgroup of bipolar patients?
Puerperal psychosis was found to be the 1st illness for 13 (36%) of a series of 36 mothers with bipolar affective disorder. The risk of recurrence with childbirth in those with established bipolar disorder was found to be 25-40%. Those women who survived childbirth without illness but then became ill at a later time reported a worse illness course. Recent fatherhood was not a precipitant of 1st affective illness for any of the 28 bipolar men.


Information technology ability mediates the association between older adults' subjective age and social activity: A STROBE-compliant cross-sectional analysis.
The association between subjective age and social activity has been reported in the extant literature, but whether this association is mediated by information technology ability and its domains (i.e., internet use assessment, packaged software use assessment, and innovativeness attitude) has not been examined. To assess the association between subjective age and social activity and to ascertain whether this association is mediated by information technology ability. This study adopted a cross-sectional design characterising sensitivity analyses and common methods bias. The participants were 895 community-dwelling older adults aged 60 years or higher in Accra, Ghana. We measured subjective age, information technology ability, and social activity with previously validated Likert scales, each of which was internally consistent at a Cronbach's α ≥0.7. The data were analysed with partial least squares structural equation modelling (PLS-SEM) and hierarchical linear regression (HLR) analysis. Subjective age was positively associated with social activity, and this association was partially mediated by information technology ability but none of the three domains of information technology ability mediated this relationship. Subjective age was positively associated with information technology ability and its three domains. Information technology ability (but not its domains) was positively associated with social activity. Older subjective age was associated with higher social activity through information technology ability. Social activity and information technology ability levels among older adults depend on subjective age, which has implications for ageing and gerontology as reported in this paper.


Flow cytometric quantitation of EpCAM-positive extracellular vesicles by immunomagnetic separation and phospholipid staining method.
Extracellular vesicles (EV) have attracted attention as circulating biomarkers for many diseases, particularly cancer. Conventional immunofluorescence staining has been used for the detection of target antigens on EV by flow cytometry. However, the staining intensity depends on the amount of antigen expressed on the vesicles and is often only around the noise level. Instead of immunofluorescence, we combined immunomagnetic separation using nanosize MACS® MicroBeads with phospholipid staining of EV (IMS-PS method). EpCAM-positive EV were prepared from the culture supernatants of OVCAR3 (EpCAM-high), A431 (EpCAM-low) or Colon-26 (non-human control) cells as cancer models and were examined by the IMS-PS method using EpCAM mAb-coated MicroBeads. By employing Polaric-500c6F as the dye for staining EV phospholipids and using appropriate flow cytometry settings, autofluorescence was excluded, whereas pretreatment of the MicroBeads with conventional blocking agents reduced nonspecific binding to non-target vesicles. These modifications resulted in a linear relation between the number of EV detected and the sample volume, regardless of the level of EpCAM expression on the vesicles. A431 EV spiked into healthy volunteer plasma were enumerated with good accuracy. The IMS-PS method may be useful for clinical evaluation of EV with low levels of antigen expression that are difficult to detect by conventional immunofluorescence.


TGF-β induces HLA-G expression through inhibiting miR-152 in gastric cancer cells.
Mounting evidences have showed the important role of transforming growth factor-β (TGF-β) in immunological surveillance of tumors. Some studies have also indicated human leukocyte antigen (HLA)-G-associated immune escape involving TGF-β management in gastric cancer (GC). However, the mechanism underlying it is unclear. This study aims to verify the correlations between HLA-G and TGF-β, involving the potential targeting of miR-152 on HLA-G. TGF-β and HLA-G levels were analyzed in blood samples from twenty GC patients with ELISA assays, while TGF-β showed directly proportional to HLA-G levels in GC patients, and TGF-β induced HLA-G up-regulation was also confirmed in GC cell lines. Furthermore, miR-152 expression could be inhibited by TGF-β, and the negative post-transcriptionally regulation of miR-152 on HLA-G was also demonstrated through gain- and loss-of-function studies. Besides, miR-152 overexpression repressed HLA-G up-regulation induced by TGF-β. And, miR-152 expression levels showed inversely proportional to both HLA-G and also TGF-β levels in GC patients. TGF-β could induce HLA-G expression in GC by inhibiting miR-152, involving its negative regulation on HLA-G. Since TGF-β induced HLA-G up-regulation plays important role in immune escape, a potential application of miR-152 was suggested in GC treatment, or miR-152 might be one potential biomarker for GC.


Neoadjuvant pertuzumab plus trastuzumab in combination with anthracycline- free chemotherapy regimen in patients with HER2 positive breast cancer-Real-world data from a single center in India.
Dual targeted therapy with chemotherapy is one of the therapeutic approaches as neoadjuvant treatment in HER2/neu positive breast cancer (BC). However, the safety and efficacy data of dual-targeted, chemotherapy regimen (docetaxel, carboplatin, trastuzumab, & pertuzumab [TCH-P] is limited from the Indian subcontinent. This retrospective study aims to evaluate the efficacy and toxicity of neoadjuvant TCH-P regimen in early, locally advanced, and oligometastatic (OM) HER2-positive BC, at All India Institute of Medical Sciences, New Delhi, India, in between the period 2015-2020. Total 6 cycles of 3-weekly neoadjuvant chemotherapy (NACT) protocol containing docetaxel (75 mg/m2), carboplatin (AUC = 6), trastuzumab (8 mg/kg loading followed by 6 mg/kg) and pertuzumab (840 mg loading followed by 420 mg) were planned. Subcutaneous peg-filgrastim was prophylactically administered on day 2 of each cycle. The primary outcome was the pathological complete response (pCR), and secondary outcomes were clinical overall response rate (ORR), rate of breast conservation surgery (BCS) for patients for whom modified radical mastectomy (MRM) was planned and toxicity. Forty-five patients with a median age of 48 years (31-65) were included in this study. The TNM (AJCC-7th edition) stage distribution was stage II, 14 (31.1%); stage III, 29 (64.5%); and stage IV (OM), 2 (4.4%). Clinical node positivity disease was found in 26 (57.8%) cases. Nineteen (42.2%) patients had hormone-positive and 26 (57.8%) cases were premenopausal. The clinical ORR and CR were seen in 100% and 60% respectively. Overall pCR rate was observed in 25 (55.6%) patients (70% in stage II). BCS was performed in 23 (51.1%) cases. In 12 (26.6%) cases, planned MRM was changed to BCS following NACT. Grade 3 and 4 toxicities were diarrhea 7 cases, thrombocytopenia in 6, neutropenia in 4, febrile neutropenia in 1, and anaemia in 2 cases. Ten patients required dose modification. No patient had congestive heart failure or induction death. This is the first study of the non-anthracycline-based neoadjuvant protocol in HER2 positive BC from India. The TCH-P is an effective, safe, and well tolerated protocol with a pCR rate of 55.6% and 26.6% BCS conversion rate from planned MRM.


Ouabain-induced increase in dopamine release from mouse striatal slices is antagonized by riluzole.
We have examined the effects of riluzole, a neuroprotective drug which stabilizes voltage-dependent sodium channels in their inactivated state and inhibits the release of glutamate in-vivo and in-vitro, on the release of newly taken up [3H]dopamine induced by ouabain, a potent and selective inhibitor of Na+/K+-ATPase in mouse striatal slices in-vitro. Riluzole potently (IC50 (concentration resulting in 50% inhibition) = 0.9+/-0.3 microM) and dose-dependently antagonized ouabain-stimulated [3H]dopamine release, the effect being observed at low concentrations. Tetrodotoxin (1 microM) and nomifensine (10 microM) also abolished ouabain-induced [3H]dopamine release. Blockade of glutamate receptors with dizocilpine (1 microM) and 6-(1H-imidazol-1-yl)-7-nitro-2,3(1H,4H)-quinoxalinedione (YM-90K; 10 microM), alone or in combination, was without effect. Incubation of striatal slices with 50 microM La3+, which blocks voltage-dependent calcium channels, did not inhibit [3H]dopamine release induced by ouabain. The potent effects of riluzole observed in this model are probably related to its ability to block voltage-dependent sodium channels. The consequences of this activity are critically discussed in relation to the protective action of riluzole previously reported in various models of Parkinson's disease and other neurodegenerative disorders.


Isolation of nonobese diabetic mouse T-cells that recognize novel autoantigens involved in the early events of diabetes.
Insulin-dependent diabetes mellitus (IDDM) is thought to result from chronic, cell-mediated, autoimmune islet damage. Our aim was to identify the earliest T-cell autoantigen in IDDM, reasoning that this antigen could be causally involved in the initiation of the disease. Identification of the earliest beta-cell-specific autoantigen is extremely important in allowing advances in prevention and treatment of initial events in the development of inflammatory insulitis that precedes beta-cell destruction and overt diabetes. Therefore, we analyzed the proliferative responses of peripheral T-cells from young, female nonobese diabetic (NOD) mice to extracts of pancreatic beta-cell lines. We were able to demonstrate that T-cells responsive to beta-cell antigens exist in the peripheral lymphoid tissue of these mice in the absence of deliberate priming before the manifestation of histologically detectable insulitis. T-cell lines and clones isolated from the peripheral lymphatic tissues of young, unimmunized, female NOD mice were also shown to react with extracts of beta-cells. Fractionation of the beta-cell extracts showed that these T-cell clones recognized multiple beta-cell-specific autoantigens but none of the previously reported putative autoantigens (glutamic acid decarboxylase [GAD]65, GAD67, Hsp65, insulin, ICA 69, carboxypeptidase-H, and peripherin). Thus, we can conclude that these responses are specific for novel beta-cell autoantigens. Finally, NOD T-cell proliferative responses were also seen to an extract of human islets suggesting potential shared antigenic determinants between human and mouse beta-cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Infection control: reducing the risk to medical patients.
Patients on medical wards are at risk of infections, both from their conditions and invasive procedures carried out on them. Scrupulous infection control by nurses can do much to reduce this risk.


Reviewing health promotion in nursing education.
The [World Health Organisation, 2000. Nurse and Midwives for Health: A WHO European Strategy for Nursing and Midwifery Education. WHO Regional Office for Europe, Copenhagen] European Strategy for Nursing and Midwifery Education calls for the explicit inclusion and application of health promotion in all nursing curricula. Prior to this strategy, and subsequently, studies have explored the nature and extent of health promotion in nursing education. This article extensively reviews this body of literature. Overall, the literature, both included in this review and its supporting discussion, presents a picture suggesting that the call for effective inclusion of health promotion has in many cases gone unheeded. The literature also identifies that the educational delivery of broader elements of health promotion is muted in comparison to the 'traditional' constructs of health education. Considerations for wider reform, born out of the literature, are presented.


A Randomized Clinical Trial Evaluating the Effects of Oligosaccharides on Transfer of Passive Immunity in Neonatal Dairy Calves.
Bacterial contamination of colostrum is common and can decrease IgG absorption in neonatal calves. Strategies that mitigate this situation without complicating colostrum management will benefit dairy calf health and survival. To evaluate the effects of supplementing colostrum with oligosaccharides (OS) on serum IgG concentration and apparent efficiency of absorption of IgG (AEA%) in calves fed unpasteurized colostrum and characterize these outcomes with respect to colostrum bacterial exposures. One hundred twenty-three neonatal dairy calves. Randomized, blinded, controlled clinical trial conducted at a commercial dairy operation. Calves were enrolled at birth in 1 of 4 treatment groups. Data were complete for 123 calves, which were distributed across the treatment groups as follows: mannan-oligosaccharides (MOS), n = 33; Saccharomyces galacto-oligosaccharides (SGOS), n = 31; Bifidobacterium galacto-oligosaccharides (BGOS), n = 28; and lactose control (CON), n = 31. A commercial radial immunodiffusion kit was used to determine colostrum and serum IgG concentrations. Conventional microbiology methods were used to enumerate colostrum bacterial counts. Bacterial counts were not significantly different among treatment groups. Total bacterial plate counts (TPC) were relatively low for the majority of colostrum samples, but TPC had a significant negative effect on serum IgG concentration and AEA% in the lactose-supplemented control group but not the OS treatment groups. These results suggest that a complement of OS structures may mitigate adverse effects of bacteria on transfer of passive immunity (TPI).


Focal foveal atrophy of unknown etiology: clinical pictures and possible underlying causes.
Focal foveal atrophy is defined as the presence of a small, focal, ill-defined, hypopigmented foveal or juxtafoveal lesion, with the remaining retina unaffected. The purpose of this study was to report the clinical characteristics and optical coherence tomography (OCT) in patients with focal foveal atrophy of unknown etiology. The study was a retrospective observational case series. Data collected included complete ocular examination results for best corrected visual acuity (BCVA), ophthalmoscopy, fundus photography, fluorescein angiography, color sense discrimination tests, visual field tests, and OCT examinations. Twenty-three eyes in 21 patients were examined. The mean patient age was 49.2 ± 15.4 years. The mean BCVA was 20/25. The 21 patients were divided into three groups according to OCT results. Group 1 eyes (n = 10) had intact inner and outer hyperreflective layers (HRLs), with the signal of the inner HRL corresponding to the junction between the inner and outer photoreceptor segments and the outer HRL corresponding to the retinal pigment epithelium (RPE). Group 2 eyes (n = 9) had small hyporeflective defects with defects in the inner HRL at the fovea but an intact outer HRL. Group 3 eyes (n = 4) had small hyporeflective defects in both the inner and outer HRLs at the fovea. Groups 3 eyes had significantly lower visual acuity compared to Group 1 eyes and Group 2 eyes. There was no significant difference in visual acuity between Group 1 and Group 2 eyes. There were no significant differences among the groups with respect to color vision or foveal thickness. This is the first report of clinical presentations for patients with focal foveal atrophy of unknown etiology. OCT aided in the diagnosis and assessment of the degree of retinal structural abnormalities, but the real etiology of foveal atrophy remains unclear.


Green preparation of carbon quantum dots with wolfberry as on-off-on nanosensors for the detection of Fe3+ and l-ascorbic acid.
A green and facile hydrothermal synthesis approach is proposed for the preparation of nitrogen-doped carbon quantum dots (N-CQDs) with wolfberry. These N-CQDs were developed as a highly sensitive fluorescent 'on-off-on' switch sensor for the sensing of Fe3+ and l-ascorbic acid (AA). The N-CQDs displayed superior fluorescence characteristics of CQDs with a quantum yield up to 22%. The N-CQDs were demonstrated to selectively react with Fe3+, leading to fluorescence quenching effect, which was successfully used for the detection of Fe3+ with a limit of detection at 3 μmoL•L-1. The addition of AA is supposed to repair the surface defects, and result in the fluorescence recovery. Based on this effect, the strategy of 'on-off-on' detection of AA was established with a limit of detection at 1.8 μmoL•L-1. Furthermore, the practical application of the detection of Fe3+ lake water and AA in medical tablet was demonstrated, promising an effective and efficient 'on-off-on' nanosensor with low-cost, green synthesis for Fe3+ and AA detection.


TDR Technique for Estimating the Intensity of Evapotranspiration of Turfgrasses.
The paper presents a method for precise estimation of evapotranspiration of selected turfgrass species. The evapotranspiration functions, whose domains are only two relatively easy to measure parameters, were developed separately for each of the grass species. Those parameters are the temperature and the volumetric moisture of soil at the depth of 2.5 cm. Evapotranspiration has the character of a modified logistic function with empirical parameters. It assumes the form ETR(θ (2.5 cm), T (2.5 cm)) = A/(1 + B · e (-C · (θ (2.5 cm) · T (2.5 cm)), where: ETR(θ (2.5 cm), T (2.5 cm)) is evapotranspiration [mm · h(-1)], θ (2.5 cm) is volumetric moisture of soil at the depth of 2.5 cm [m(3) · m(-3)], T (2.5 cm) is soil temperature at the depth of 2.5 cm [°C], and A, B, and C are empirical coefficients calculated individually for each of the grass species [mm · h(1)], and [-], [(m(3) · m(-3) · °C)(-1)]. The values of evapotranspiration calculated on the basis of the presented function can be used as input data for the design of systems for the automatic control of irrigation systems ensuring optimum moisture conditions in the active layer of lawn swards.


Prestress as an optimal biomechanical parameter for needle penetration.
Drug delivery requires precise intradermal and subcutaneous injections of formulations to clinically relevant penetration depths. However, penetration depth is confounded by skin deflection, which occurs prior to and during penetration as the skin surface deforms axially with the needle, and which varies profoundly due to differing intrinsic mechanical (e.g. viscoelastic) tissue properties, disease state, aging, and ethnicity. Herein, an ex vivo model was utilized to study factors that affect skin deflection and the efficacy of injection, including prestress applied at the tissue surface, needle gauge, velocity, and actuation depth. The application of prestress minimized skin deflection during needle penetration and allowed for needle actuation to the targeted penetration depths with minimum variability. The force required to achieve target penetration depths was found to increase with prestress and decrease with needle gauge. Our findings emphasize the need for prestress applied to the skin surface to minimize variation in skin properties and administer formulations for intradermal and subcutaneous treatments with maximum precision.


Pregnancy-induced hypertension decreases Kv1.3 potassium channel expression and function in human umbilical vein smooth muscle.
Voltage-gated potassium (Kv) channels are the largest superfamily of potassium (K) channels. A variety of Kv channels are expressed in the vascular smooth muscle cells (SMC). Studies have shown that gestational diabetes mellitus (GDM) and pregnancy-induced hypertension (PIH) cause various changes in the human umbilical vein (HUV). Recently, we have shown that 4-AP, a nonspecific Kv1-4 channel inhibitor, significantly decreases vasorelaxation induced by K channel opener pinacidil in vascular SMCs of the HUVs from normal pregnancies, but not in GDM and PIH. The goal of this study was to provide more detailed insight in the Kv channel subtypes involved in pinacidil-induced vasodilation of HUVs, as well as to investigate potential alterations of their function and expression during GDM and PIH. Margatoxin, a specific blocker of Kv1.2 and Kv1.3 channels, significantly antagonized pinacidil-induced vasorelaxation in normal pregnancy, while in HUVs from GDM and PIH that was not the case, indicating damage of Kv1.2 and Kv1.3 channel function. Immunohistochemistry and Western blot revealed similar expression of Kv1.2 channels in all groups. The expression of Kv1.3 subunit was significantly decreased in PIH, while it remained unchanged in GDM compared to normal pregnancy. Phrixotoxin, specific blocker of Kv4.2 and Kv4.3 channels, did not antagonize response to pinacidil in any of the groups. The major novel findings show that margatoxin antagonized pinacidil-induced relaxation in normal pregnancy, but not in GDM and PIH. Decreased expression of Kv1.3 channels in HUV during PIH may be important pathophysiological mechanism contributing to an increased risk of adverse pregnancy outcomes.


Learning new sounds of speech: reallocation of neural substrates.
Functional magnetic resonance imaging (fMRI) was used to investigate changes in brain activity related to phonetic learning. Ten monolingual English-speaking subjects were scanned while performing an identification task both before and after five sessions of training with a Hindi dental-retroflex nonnative contrast. Behaviorally, training resulted in an improvement in the ability to identify the nonnative contrast. Imaging results suggest that the successful learning of a nonnative phonetic contrast results in the recruitment of the same areas that are involved during the processing of native contrasts, including the left superior temporal gyrus, insula-frontal operculum, and inferior frontal gyrus. Additionally, results of correlational analyses between behavioral improvement and the blood-oxygenation-level-dependent (BOLD) signal obtained during the posttraining Hindi task suggest that the degree of success in learning is accompanied by more efficient neural processing in classical frontal speech regions, and by a reduction of deactivation relative to a noise baseline condition in left parietotemporal speech regions.


Single clonal origin of neoplastic B-cells with different immunoglobulin light chains in a patient with Richter's syndrome.
A 71-year-old man was found to have chronic lymphocytic leukemia (CLL) and diffuse large cell lymphoma (DLC) simultaneously and was diagnosed as Richter's syndrome. The CLL had mu lambda surface immunoglobulin (sIg) whereas the DLC had mu kappa sIg. However, the immunoglobulin (Ig) gene rearrangement and surface marker analysis demonstrated that both CLL and DLC had identical rearrangement patterns of the Ig heavy chain (IgH) and identical surface markers CD5+, CD19+, and CD20+. These facts imply that in this case the two malignancies are of single clonal origin initially, and that different sIg of CLL and DLC do not, therefore, necessarily indicate the biclonality of these malignancies. The origin of DLC in Richter's syndrome remains controversial. This case suggests difficulty in concluding the biclonality of these malignancies. For a conclusion on clonality to be definitive, there is a need for cloning and nucleotide sequencing of rearranged Ig genes in more patients with Richter's syndrome.


Initial Evaluation of Therapy Response after Adjuvant Radioiodine Therapy in Patients with Early-Stage Papillary Thyroid Cancer-Does Time Matter?
The aim was to assess ablation success after initial radioiodine (RAI) therapy in early-stage PTC patients and compare outcomes of first diagnostic control after 6 and 9 months (6m/9m-DC) to examine whether time could possibly avoid unnecessary overtreatment. There were 353 patients who were matched regarding age, sex, and tumor stage and divided in two groups depending on time of first DC (6m- and 9m-DC). Therapy response was defined as thyroglobulin level <0.5 ng/mL, no pathological uptake in the diagnostic I-131 whole-body scintigraphy (WBS), and no further RAI therapy courses. The 6m-DC group was further divided into endogenously and exogenously stimulated TSH before RAI therapy and compared regarding outcome. No significant differences were found between 6m-DC vs. 9m-DC regarding I-131 uptake in WBS (p = n.s.), Tg levels (p = n.s.), re-therapy rates (p = n.s.), and responder rates (p = n.s.). Significantly less relevant pathological I-131 uptake was found in WBS (p = 0.006) in endogenously compared to exogenously stimulated 6m-DC patients, resulting in lower re-therapy (p = 0.028) and higher responder rates (p = 0.001). DC at 6 months after RAI therapy and stimulation with recombinant human thyroid-stimulating hormone (rhTSH) represent the most balanced solution. Particularly regarding quality of life and mental relief of patients, early DC with rhTSH represents sufficient and convenient assessment of ablation success.


Remote Controlled Autonomous Microgravity Lab Platforms for Drug Research in Space.
Research conducted in microgravity conditions has the potential to yield new therapeutics, as advances can be achieved in the absence of phenomena such as sedimentation, hydrostatic pressure and thermally-induced convection. The outcomes of such studies can significantly contribute to many scientific and technological fields, including drug discovery. This article reviews the existing traditional microgravity platforms as well as emerging ideas for enabling microgravity research focusing on SpacePharma's innovative autonomous remote-controlled microgravity labs that can be launched to space aboard nanosatellites to perform drug research in orbit. The scientific literature is reviewed and examples of life science fields that have benefited from studies in microgravity conditions are given. These include the use of microgravity environment for chemical applications (protein crystallization, drug polymorphism, self-assembly of biomolecules), pharmaceutical studies (microencapsulation, drug delivery systems, behavior and stability of colloidal formulations, antibiotic drug resistance), and biological research, including accelerated models for aging, investigation of bacterial virulence , tissue engineering using organ-on-chips in space, enhanced stem cells proliferation and differentiation.


Human blood dendritic cells from allergic subjects have impaired capacity to produce interferon-alpha via Toll-like receptor 9.
High-affinity IgE receptor (Fc epsilon RI) expression on blood dendritic cells reportedly correlates with serum IgE levels. Our studies demonstrate that plasmacytoid dendritic cells (pDCs) secrete pro-inflammatory cytokines (IL-6, TNF-alpha) following Fc epsilon RI stimulation - a mode of activation that simultaneously reduces expression of Toll-like receptor 9 (TLR9). Whether or not TLR9 and/or Fc epsilon RI levels and their function on dendritic cells relate to allergic status is unknown. The aim of this study is to compare the innate (TLR9-mediated) immune response of human pDCs to TLR9 and Fc epsilon RI alpha receptor expression in allergic and non-allergic subjects. Basophil-depleted mononuclear cell fractions containing pDCs were prepared from peripheral blood of allergic and non-allergic subjects. Intracellular TLR9 and surface Fc epsilon RI alpha expression in blood dendritic cell antigen-2-positive cells were determined by flow cytometry. Activating anti-IgE antibody, anti-Fc epsilon RI alpha antibody, and TLR9 agonist were used to stimulate cell suspensions, with cytokine levels determined by ELISA. No difference in the frequency of pDCs was detected among allergic (n=9) vs. non-allergic (n=11) subjects (P=0.261). While there was also no difference in the baseline expression of TLR9, pDCs from allergic subjects produced sixfold less IFN-alpha when stimulated with CpG (P=0.002). Conversely, there was higher Fc epsilon RI alpha expression (P=0.01) on the pDCs of allergic subjects. Impaired TLR9-dependent immune responses in human pDCs are associated with allergic status and inversely correlated with Fc epsilon RI alpha expression. This impaired innate immune response among dendritic cells of allergic subjects may lead to more targeted therapeutic approaches and could provide a better understanding of the mechanisms underlying conventional and CpG-based immunotherapy.


Dynamics of complex-valued fractional-order neural networks.
The dynamics of complex-valued fractional-order neuronal networks are investigated, focusing on stability, instability and Hopf bifurcations. Sufficient conditions for the asymptotic stability and instability of a steady state of the network are derived, based on the complex system parameters and the fractional order of the system, considering simplified neuronal connectivity structures (hub and ring). In some specific cases, it is possible to identify the critical values of the fractional order for which Hopf bifurcations may occur. Numerical simulations are presented to illustrate the theoretical findings and to investigate the stability of the limit cycles which appear due to Hopf bifurcations.


Growth inhibition of subcutaneous mouse melanoma and induction of natural killer cells by liposome-mediated interferon-beta gene therapy.
In this study we investigated the antitumour effect and mechanism of action of cationic liposome-mediated murine interferon-beta (IFNbeta) gene therapy in mouse B16F1 melanoma cells in vitro and in vivo. Murine IFNbeta gene transfer by cationic liposome resulted in substantial growth inhibition of B16F1 melanoma cells in culture when compared with phosphate buffered saline or recombinant murine IFNbeta treatment, or lacZ control gene transfer. Use of video-enhanced contrast-differential interference contrast (VEC-DIC) microscopy revealed that liposomes containing the murine IFNbeta gene [lip(pSV2muIFNbeta)], but not recombinant murine IFNbeta, induced dramatic morphological changes that characterize apoptosis, including bleb formation, shrinkage of cells, nuclear condensation and 'ballooning', in approximately 30% of the cells treated. Intratumoral administration of lip(pSV2muIFNbeta) resulted in a 5.5-fold reduction in the mean volume of subcutaneous melanoma lesions in syngeneic mice 15 days after treatment and eradicated the tumour in 18% of the mice treated. Immunocytochemical analysis demonstrated that a larger number of natural killer (NK) cells infiltrated the tumour following lip(pSV2muIFNbeta) treatment than in controls. In vivo depletion of NK cells using the anti-asialoGM1 antibody reduced the efficacy of lip(pSV2muIFNbeta) treatment. Taken together, our data suggest that cationic liposome-mediated IFNbeta gene therapy could be effective against melanoma by directly inducing cell death and stimulating NK cells.


Immediate extended latissimus dorsi flap reconstruction after skin-sparing mastectomy for breast cancer associated with paraffinoma: report of a case.
Paraffin oil injection into the breast, which had been used for breast augmentation in the past, can cause various complications. Complete removal of paraffinoma lesions with immediate breast reconstruction using autogenous tissue is a very satisfactory treatment option. However, diffuse random distribution of paraffin oil through the subdermal layer makes it impossible to remove all of the lesions with overlying skin without resulting in a shortage of available skin and poor cosmesis. We herein report the case of a patient with breast cancer associated with paraffinoma, treated with skinsparing mastectomy and axillary node dissection with immediate extended latissimus dorsi flap reconstruction, resulting in good cosmetic outcome, and showing no complications during the wound-healing process.


Variability in agreement between physicians and nurses when measuring the Glasgow Coma Scale in the emergency department limits its clinical usefulness.
To assess the interrater reliability of the Glasgow Coma Scale (GCS) between nurses and senior doctors in the ED. This was a prospective observational study with a convenience sample of patients aged 18 or above who presented with a decreased level of consciousness to a tertiary hospital ED. A senior ED doctor (emergency physicians and trainees) and registered nurse each independently scored the patient's GCS in blinded fashion within 15 min of each other. The data were then analysed to determine interrater reliability using the weighted kappa statistic and the size and directions of differences between paired scores were examined. A total of 108 eligible patients were enrolled, with GCS scores ranging from 3 to 14. Interrater agreement was excellent (weighted kappa > 0.75) for verbal scores and total GCS scores, and intermediate (weighted kappa 0.4-0.75) for motor and eye scores. Total GCS scores differed by more than two points in 10 of the 108 patients. Interrater agreement did not vary substantially across the range of actual numeric GCS scores. Although the level of agreement for GCS scores was generally high, a significant proportion of patients had GCS scores which differed by two or more points. This degree of disagreement indicates that clinical decisions should not be based solely on single GCS scores.


Stepwise strategy to improve Cervical Cancer Screening Adherence (SCAN-CC): automated text messages, phone calls and face-to-face interviews: protocol of a population-based randomised controlled trial.
Screening is highly effective for cervical cancer prevention and control. Population-based screening programmes are widely implemented in high-income countries, although adherence is often low. In Portugal, just over half of the women adhere to cervical cancer screening, contributing for greater mortality rates than in other European countries. The most effective adherence raising strategies are based on patient reminders, small/mass media and face-to-face educational programmes, but sequential interventions targeting the general population have seldom been evaluated. The aim of this study is to assess the effectiveness of a stepwise approach, with increasing complexity and cost, to improve adherence to organised cervical cancer screening: step 1a-customised text message invitation; step 1b-customised automated phone call invitation; step 2-secretary phone call; step 3-family health professional phone call and face-to-face appointment. A population-based randomised controlled trial will be implemented in Portuguese urban and rural areas. Women eligible for cervical cancer screening will be randomised (1:1) to intervention and control. In the intervention group, women will be invited for screening through text messages, automated phone calls, manual phone calls and health professional appointments, to be applied sequentially to participants remaining non-adherent after each step. Control will be the standard of care (written letter). The primary outcome is the proportion of women adherent to screening after step 1 or sequences of steps from 1 to 3. The secondary outcomes are: proportion of women screened after each step (1a, 2 and 3); proportion of text messages/phone calls delivered; proportion of women previously screened in a private health institution who change to organised screening. The intervention and control groups will be compared based on intention-to-treat and per-protocol analyses. The study was approved by the Ethics Committee of the Northern Health Region Administration and National Data Protection Committee. Results will be disseminated through communications in scientific meetings and peer-reviewed journals. NCT03122275.


Early versus late surgical intervention for central cord syndrome: A nationwide all-payer inpatient analysis of length of stay, discharge destination and cost of care.
Central cord syndrome (CCS) is the most common incomplete spinal cord injury (SCI), resulting in various degrees of neurologic compromise below the level of the affected cervical cord. The management of CCS is controversial regarding not only whether to surgically intervene, but also when surgery should occur. In this study, we conduct the largest multi-center study to date examining differences in inpatient outcomes, general discharge disposition, length of stay, and cost associated with early versus late surgical intervention for CCS. The National Inpatient Sample (NIS) was queried for years 2012-2015 for patients who underwent surgery with a primary diagnosis of CCS. The median interval between admission and intervention was noted. Patients operated upon prior to this timepoint were placed in the early surgery group, and others into the later surgery group. The groups were then compared, while using 1:1 propensity score matching to control for baseline presentation, with respect to mortality, discharge disposition, length of stay, and total charges. A total of 422 patients met inclusion and exclusion criteria. The median time from admission to intervention was 2 days. Patients with higher initial severity of injury were more likely to undergo early surgery. Upon controlling for severity of initial presentation, earlier intervention did not appear to affect mortality or post-operative length of stay. However, patients operated upon earlier had more favorable discharge destinations (p = 0.025) and a lower associated cost of care ($198,050.70 vs. $243,048.10, p = 0.009). Earlier surgical intervention for CCS may result in better patient disposition and less total charges. III.


miR-15b and miR-16 regulate TNF mediated hepatocyte apoptosis via BCL2 in acute liver failure.
Acute liver failure (ALF) still has an unacceptable high mortality rate, despite substantial improvements with multidisciplinary care. The precise underlying mechanism of ALF remains to be explored. It has been reported that microRNAs (miRNAs) are novel regulators in a number of liver diseases, but the role of miRNAs in the development of ALF is not fully understood. An ALF murine model was generated by ip injection of D: -GalN/LPS, which was confirmed with histopathology and biochemistry. The hepatic miRNA expression profile in ALF was determined by microarray and verified by qRT-PCR. The functions and signal pathways of the targeted genes of these deregulated miRNAs were predicted, using bioinformatics analysis. The possible underlying mechanism was investigated by exploring the relationship between miRNA modification and hepatocyte apoptosis. There were a total of 95 significantly changed miRNAs in ALF compared to mock-treated (P < 0.01). Among these 95 miRNAs, 20 were up-regulated and 26 were down-regulated at both 5 and 7 h time points. Bioinformatics analysis predicted that some of these 46 miRNAs were involved in apoptosis. Among the up-regulated miRNAs involved in apoptosis, miR-15b and miR-16 showed the highest enrichment and targeted the common anti-apoptotic gene, BCL2. Our in vitro data demonstrated that miR-15b and/or miR-16 regulated BCL2 at the protein level. Inhibition of miR-15b and/or miR-16 reduced hepatic apoptosis and TNF production. These data suggest that miR-15b and miR-16 regulate TNF mediated hepatic apoptosis via BCL2 during ALF, and may shed light on the development of a therapeutic strategy for treatment of ALF.


Kindlin-2 protects pancreatic β cells through inhibiting NLRP3 inflammasome activation in diabetic mice.
Diabetes mellitus has been a major public health problem worldwide, characterized by insulin resistance and dysfunction of β-cells. A previous study showed that Kindlin-2 loss in β-cells dramatically reduces insulin secretion and decreases β-cell mass, resulting in severe diabetes-like phenotypes. It suggests that Kindlin-2 in β-cells play an important role in regulating glucose homeostasis. However, the effect of Kindlin-2 on the function of β-cells under chronic hyperglycemia in diabetes has not been explored. Here we report that Kindlin-2 overexpression ameliorates diabetes and improves insulin secretion in mice induced by streptozocin. In contrast, Kindlin-2 insufficiency exacerbates diabetes and promotes β-cells dysfunction and inflammation in β-cells induced by a high-fat diet (HFD). In vitro, Kindlin-2 overexpression prevented high-glucose (HG)-induced dysfunction in β-cells. Kindlin-2 overexpression also decreased the expression of pro-inflammatory cytokines and NLRP3 inflammasome expression in β-cells exposed to HG. Furthermore, the loss of Kindlin-2 aggravates the expression of inflammatory cytokines and NLRP3 induced by HG in β-cells. Collectively, we demonstrate that Kindlin-2 protects against diabetes by inhibiting NLRP3 inflammasome activation.


Pharmacological cognitive enhancement: treatment of neuropsychiatric disorders and lifestyle use by healthy people.
Neuropsychiatric disorders typically manifest as problems with attentional biases, aberrant learning, dysfunctional reward systems, and an absence of top-down cognitive control by the prefrontal cortex. In view of the cost of common mental health disorders, in terms of distress to the individual and family in addition to the financial cost to society and governments, new developments for treatments that address cognitive dysfunction should be a priority so that all members of society can flourish. Cognitive enhancing drugs, such as cholinesterase inhibitors and methylphenidate, are used as treatments for the cognitive symptoms of Alzheimer's disease and attention deficit hyperactivity disorder. However, these drugs and others, including modafinil, are being increasingly used by healthy people for enhancement purposes. Importantly for ethical and safety reasons, the drivers for this increasing lifestyle use of so-called smart drugs by healthy people should be considered and discussions must occur about how to ensure present and future pharmacological cognitive enhancers are used for the benefit of society.


Regulation of immune cell adhesion and migration by regulator of adhesion and cell polarization enriched in lymphoid tissues.
Rap1 has emerged as an important regulator of adhesion in multicellular organisms. In the immune system, Rap1 functions as an inside-out signalling molecule for leucocyte integrins following stimulation with chemokines or antigens. Regulator of adhesion and cell polarization enriched in lymphoid tissues (RAPL) is a novel Rap1 effector molecule that mediates Rap1 signalling to integrins. The Rap1-RAPL complex regulates the spatial distribution of the integrin lymphocyte function-associated antigen-1 as well as cell polarization. The linking of inside-out signalling with polarization synergistically promotes highly efficient lymphocyte trafficking. Targeted deletion of RAPL in mice has demonstrated multiple indispensable roles for this protein in lymphocyte and dendritic cell trafficking critical for immunosurveillance.


Adjunctive Pimavanserin in Patients with Major Depressive Disorder: Combined Results from Two Randomized, Double-Blind, Placebo-Controlled Phase 3 Studies.
In a phase 2 study, pimavanserin demonstrated efficacy as adjunctive treatment for major depressive disorder (MDD). Subsequently, two phase 3 studies (NCT03968159 in the US; NCT03999918 in Europe) were initiated to examine the efficacy and safety of adjunctive pimavanserin in subjects with MDD and inadequate response to antidepressant treatment. Studies were combined with a prespecified statistical analysis plan owing to recruitment challenges related to the COVID-19 pandemic. The randomized, double-blind studies enrolled 298 patients with MDD and inadequate response to current antidepressants. Patients were randomly assigned 1:1 to pimavanserin or placebo added to current antidepressant for 6 weeks. Primary endpoint was change from baseline to week 5 in the Hamilton Rating Scale for Depression, 17-item version (HAM-D-17). There was no effect of pimavanserin in change from baseline to week 5 in the HAM-D-17 (pimavanserin [n = 138]: least-squares mean [LSM] [standard error {SE}], -9.0 [0.58]; placebo [n = 135]: -8.1 [0.58]; mixed-effects model for repeated measures LSM [SE] difference, -0.9 [0.82], P = 0.2956). Nominal improvement with pimavanserin was observed on 2 secondary endpoints: Clinical Global Impressions-Severity scale, Karolinska Sleepiness Scale. Treatment-emergent adverse events occurred in 58.1% of pimavanserin-treated and 54.7% of placebo-treated patients. Adjunctive pimavanserin did not significantly improve depressive symptoms, although pimavanserin was well tolerated.


Time-space trade-offs in population protocols for the majority problem.
Population protocols are a model for distributed computing that is focused on simplicity and robustness. A system of n identical agents (finite state machines) performs a global task like electing a unique leader or determining the majority opinion when each agent has one of two opinions. Agents communicate in pairwise interactions with randomly assigned communication partners. Quality is measured in two ways: the number of interactions to complete the task and the number of states per agent. We present protocols for the majority problem that allow for a trade-off between these two measures. Compared to the only other trade-off result (Alistarh et al. in Proceedings of the 2015 ACM symposium on principles of distributed computing, Donostia-San Sebastián, 2015), we improve the number of interactions by almost a linear factor. Furthermore, our protocols can be made uniform (working correctly without any information on the population size n), yielding the first uniform majority protocols that stabilize in a subquadratic number of interactions.