A plasmid that replicates in both mouse and E. coli cells.
Cloned bovine papilloma virus (BPV) DNA induces cellular transformation when introduced into mouse cells growing in culture; the transferred viral DNA replicates as an extrachromosomal, closed circular element. BPV DNA is therefore an inviting replicon to construct a "shuttle" vector that can replicate in both mammalian cells and E. coli. Although BPV DNA devoid of bacterial plasmid sequences has been successfully employed to reintroduce cloned genes into rodent cells, construction of a true shuttle plasmid has been hampered by a disruptive influence of bacterial plasmid sequences that appear to block cellular transformation when included on the transferred molecule. We constructed a molecule, pGP, containing the transforming region of the BPV genome, the rat growth hormone gene, and bacterial plasmid pBR 327, and have found unexpectedly that the intact molecule can induce cellular transformation of mouse cells at high efficiency despite the presence of bacterial sequences in the transferred plasmid. A similar plasmid without the growth hormone segment does not transform mouse cells. The pGP molecule replicates as a stable plasmid in both mouse cells, where there are 30 to 80 monomer episomes per cell, and E. coli, and may be shuttled back and forth unaltered between the two kinds of cells. The growth hormone gene is transcribed in the mouse cells and gives rise to a transcript that is longer than authentic rat growth hormone mRNA and does not appear to be regulated by glucocorticoids. When pGP is cotransferred into mouse L-cells with herpes simplex virus tk gene, it appears to integrate, and free monomer episomes are not observed.


Microglial Phagocytosis of Neurons: Diminishing Neuronal Loss in Traumatic, Infectious, Inflammatory, and Autoimmune CNS Disorders.
Errors in neuron-microglial interaction are known to lead to microglial phagocytosis of live neurons and excessive neuronal loss, potentially yielding poorer clinical outcomes. Factors that affect neuron-microglial interaction have the potential to influence the error rate. Clinical comorbidities that unfavorably impact neuron-microglial interaction may promote a higher rate of neuronal loss, to the detriment of patient outcome. This paper proposes that many common, clinically modifiable comorbidities have a common thread, in that they all influence neuron-microglial interactions. Comorbidities like traumatic brain injury, infection, stress, neuroinflammation, loss of neuronal metabolic integrity, poor growth factor status, and other factors, all have the potential to alter communication between neurons and microglia. When this occurs, microglial phagocytosis of live neurons can increase. In addition, microglia can shift into a morphological form in which they express major histocompatibility complex II (MHC-II), allowing them to function as antigen presenting cells that present neuronal debris as antigen to invading T cells. This can increase risk for the development of CNS autoimmunity, or can exacerbate existing CNS autoimmunity. The detrimental influence of these comorbidities has the potential to contribute to the mosaic of factors that determine patient outcome in some CNS pathologies that have neuropsychiatric involvement, including TBI and CNS disorders with autoimmune components, where excessive neuronal loss can yield poorer clinical outcomes. Recognition of the impact of these comorbidities may contribute to an understanding of the common clinical observation that many seemingly disparate factors contribute to the overall picture of case management and clinical outcome in these complex disorders. In a clinical setting, knowing how these comorbidities can influence neuron-microglial interaction can help focus surveillance and care on a broader group of potential therapeutic targets. Accordingly, an interest in the mechanisms underlying the influence of these factors on neuron-microglial interactions is appropriate. Neuron-microglial interaction is reviewed, and the various mechanisms by which these potential comorbidities influence neuro-microglial interaction are described.


Reciprocal relationships between parenting behavior and disruptive psychopathology from childhood through adolescence.
Theoretical models suggest that child behaviors influence parenting behaviors, and specifically that unpleasant child behaviors coerce parents to discontinue engaging in appropriate discipline. This study examined reciprocal relationships between parenting behaviors (supervision, communication, involvement, timid discipline and harsh punishment) and child disruptive disorder symptoms (ADHD, ODD and CD) in a clinic-referred sample of 177 boys. Annual measures, including structured clinical interviews, were obtained from the beginning of the study (when boys were between the ages of 7 to 12) to age 17. Specific reciprocal influence was observed; only timid discipline predicted worsening behavior, namely ODD symptoms, and ODD symptoms predicted increases in timid discipline. Greater influence from child behaviors to parenting practices was found: ODD also predicted poorer communication and decreased involvement, and CD predicted poorer supervision. ADHD was neither predictive of, nor predicted by, parenting behaviors. The results are specifically supportive of a coercive process between child behaviors and parenting behaviors, and generally suggestive of greater influence of child behaviors on parenting behaviors than of parenting behaviors on child behaviors.


Headache in the Older Population: Causes, Diagnoses, and Treatments.
Primary headaches are less common and differ in presentation in older versus younger individuals. Secondary headaches become more common among older patients. Diagnosis and management of headaches in those > 65 years are discussed. Migraine and tension-type headaches are rarely new onset in this age group and should be a diagnosis of exclusion. In older individuals, migraine is more likely to be bilateral with less sensory sensitivities. Migraine aura may present without headache; careful assessment is needed to exclude stroke. Other primary headaches discussed include cough, hypnic, and other headaches. Secondary causes discussed include giant cell arteritis, trigeminal post-herpetic neuropathy, sleep apnea, cardiac cephalgia, cervicogenic pain, vascular etiologies, medications, and burning-mouth syndrome. In older individuals, primary headaches are diagnoses of exclusion, and treatment is affected by comorbidities and polypharmacy. Secondary headaches are a major consideration requiring appropriate workup. Many treatments can safely be offered regardless of age.


Antioxidant supplementation ameliorates molecular deficits in Smith-Lemli-Opitz syndrome.
Smith-Lemli-Opitz syndrome (SLOS) is an inborn error of cholesterol biosynthesis characterized by diminished cholesterol and increased 7-dehydrocholesterol (7-DHC) levels. 7-Dehydrocholesterol is highly reactive, giving rise to biologically active oxysterols. 7-DHC-derived oxysterols were measured in fibroblasts from SLOS patients and an in vivo SLOS rodent model using high-performance liquid chromatography tandem mass spectrometry. Expression of lipid biosynthesis genes was ascertained by quantitative polymerase chain reaction and Western blot. The effects of an antioxidant mixture of vitamin A, coenzyme Q10, vitamin C, and vitamin E were evaluated for their potential to reduce formation of 7-DHC oxysterols in fibroblast from SLOS patients. Finally, the effect of maternal feeding of vitamin E enriched diet was ascertained in the brain and liver of newborn SLOS mice. In cultured human SLOS fibroblasts, the antioxidant mixture led to decreased levels of the 7-DHC-derived oxysterol, 3β,5α-dihydroxycholest-7-en-6-one. Furthermore, gene expression changes in SLOS human fibroblasts were normalized with antioxidant treatment. The active ingredient appeared to be vitamin E, as even at low concentrations, it significantly decreased 3β,5α-dihydroxycholest-7-en-6-one levels. In addition, analyzing a mouse SLOS model revealed that feeding a vitamin E enriched diet to pregnant female mice led to a decrease in oxysterol formation in brain and liver tissues of the newborn Dhcr7-knockout pups. Considering the adverse effects of 7-DHC-derived oxysterols in neuronal and glial cultures and the positive effects of antioxidants in patient cell cultures and the transgenic mouse model, we believe that preventing formation of 7-DHC oxysterols is critical for countering the detrimental effects of DHCR7 mutations.


Ribosomal protein S6 kinase beta-1 gene variants cause hypertrophic cardiomyopathy.
Hypertrophic cardiomyopathy (HCM) is a genetic heart muscle disease with preserved or increased ejection fraction in the absence of secondary causes. Mutations in the sarcomeric protein-encoding genes predominantly cause HCM. However, relatively little is known about the genetic impact of signalling proteins on HCM. Here, using exome and targeted sequencing methods, we analysed two independent cohorts comprising 401 Indian patients with HCM and 3521 Indian controls. We identified novel variants in ribosomal protein S6 kinase beta-1 (RPS6KB1 or S6K1) gene in two unrelated Indian families as a potential candidate gene for HCM. The two unrelated HCM families had the same heterozygous missense S6K1 variant (p.G47W). In a replication association study, we identified two S6K1 heterozygotes variants (p.Q49K and p.Y62H) in the UK Biobank cardiomyopathy cohort (n=190) compared with matched controls (n=16 479). These variants are neither detected in region-specific controls nor in the human population genome data. Additionally, we observed an S6K1 variant (p.P445S) in an Arab patient with HCM. Functional consequences were evaluated using representative S6K1 mutated proteins compared with wild type in cellular models. The mutated proteins activated the S6K1 and hyperphosphorylated the rpS6 and ERK1/2 signalling cascades, suggesting a gain-of-function effect. Our study demonstrates for the first time that the variants in the S6K1 gene are associated with HCM, and early detection of the S6K1 variant carriers can help to identify family members at risk and subsequent preventive measures. Further screening in patients with HCM with different ethnic populations will establish the specificity and frequency of S6K1 gene variants.


Clinical outcomes following proton therapy for children with central nervous system tumors referred overseas.
International, multidisciplinary care of children with central nervous system (CNS) tumors presents unique challenges. The aim of this study is to report patient outcomes of U.K. children referred for proton therapy to a North American facility. From 2008 to 2016, 166 U.K. children with approved CNS tumors were treated with proton therapy at a single academic medical center in the United States. Median age was 7 years (range, 1-19). Median follow-up was 2.6 years. The 3-year actuarial overall survival (OS) and local control (LC) rates were 96% and 91%, respectively, for the overall group, 92% and 85% for the ependymoma subgroup (n = 57), 95% and 88% for the low-grade glioma subgroup (n = 54), and 100% and 100%, respectively, for the craniopharyngioma subgroup (n = 45). Cyst expansion was observed in 13 patients, including one case resulting in visual impairment. Serious side effects included new-onset seizures in three patients (1.8%), symptomatic vasculopathy in three patients (1.8%), and symptomatic brainstem necrosis in one patient (0.6%). In this cohort of British children referred overseas for proton therapy, disease control does not appear compromised, toxicity is acceptable, and improvement in long-term function is anticipated in survivors owing to the reduced brain exposure afforded by proton therapy.


Expression and activity of microsomal epoxide hydrolase in follicles isolated from mouse ovaries.
Microsomal epoxide hydrolase (mEH) is involved in the detoxification of xenobiotics that are or can form epoxide metabolites, including the ovotoxicant, 4-vinylcyclohexene (VCH). This industrial chemical is bioactivated by hepatic CYP450 to the diepoxide metabolite, VCD, which destroys mouse small preantral follicles (F1). Since ovarian mEH may play a role in VCD detoxification, these studies investigated the expression and activity of mEH in isolated ovarian fractions. Mice were given 1 or 15 daily doses (ip) of VCH (7.4 mmol/kg/day) or VCD (0.57 mmol/kg/day); 4 h following the final dose, ovaries were removed, distinct populations of intact follicles (F1, 25-100 microm; F2, 100-250 microm; F3, > 250 microm) and interstitial cells (Int) were isolated, and total RNA and protein were extracted. Real-time polymerase chain reaction and the substrate cis-stilbene oxide (CSO; 12.5 microM) were used to evaluate expression and specific activity of mEH, respectively. Confocal microscopy evaluated ovarian distribution of mEH protein. Expression of mRNA encoding mEH was increased in F1 (410 +/- 5% VCH; 292 +/- 5% VCD) and F2 (1379 +/- 4% VCH; 381 +/- 11% VCD) follicles following repeated dosing with VCH or VCD. Catalytic activity of mEH increased in F1 follicles following repeated dosing with VCH/VCD (381 +/- 11% VCH; 384 +/- 27% VCD). Visualized by confocal microscopy, mEH protein was distributed throughout the ovary with the greatest staining intensity in the interstitial cells and staining in the theca cells that was increased by dosing (56 +/- 0.8% VCH; 29 +/- 0.9% VCD). We conclude that mEH is expressed and is functional in mouse ovarian follicles. Additionally,in vivo dosing with VCH and VCD affects these parameters.


Model predictions of increased knee joint loading in regions of thinner articular cartilage after patellar tendon adhesion.
Patellar tendon adhesion is a complication from anterior cruciate ligament (ACL) reconstruction that may affect patellofemoral and tibiofemoral biomechanics. A computational model was used to investigate the changes in knee joint mechanics due to patellar tendon adhesion under normal physiological loading during gait. The calculations showed that patellar tendon adhesion up to the level of the anterior tibial plateau led to patellar infera, increased patellar flexion, and increased anterior tibial translation. These kinematic changes were associated with increased patellar contact force, a distal shift in peak patellar contact pressure, a posterior shift in peak tibial contact pressure, and increased peak tangential contact sliding distance over one gait cycle (i.e., contact slip). Postadhesion, patellar and tibial contact locations corresponded to regions of thinner cartilage. The predicted distal shift in patellar contact was in contrast to other patellar infera studies. Average patellar and tibial cartilage pressure did not change significantly following patellar tendon adhesion; however, peak medial tibial pressure increased. These results suggest that changes in peak tibial cartilage pressure, contact slip, and the migration of contact to regions of thinner cartilage are associated with patellar tendon adhesion and may be responsible for initiating patellofemoral pain and knee joint structural damage observed following ACL reconstruction.


A mutator factor in a strain of Drosophila melanogaster: identified by use of mutation, reversion rates and male recombination.
A set of 1,000 "mutation accumulation" lines Drosophila melanogaster, which originated from two different wild-type, lethal-bearing second chromosomes (Yamaguchi and Mukai 1974; Mukai and Cockerham 1977), was examined for evidence of a mutator factor by using the occurrence of recessive visible mutations and male recombination to identify its presence. The 1,000 lines were screened at approximately generation 240 for the presence of recessive visible mutation at twelve loci, by outcrossing to a balanced multiply marked second chromosome stock (Muller's "12ple" Bowling Green). Twenty-three lines were found to carry a visible mutation at one of the vg locus. mutations found in three lines, two at the dp locus and one at the vg locus, demonstrated instability as revertants to the wild type and were recovered and verified in these three cases. The three revertant lines, and three lines showing no reversion, were tested for their ability to induce male recombination. Male recombination was observed in the three lines in which revertants were recovered. Male and female sterility assays indicated conclusively that these "hybrid dysgenic" characteristics could not be used to identify lines potentially carrying mutator factors, whereas the consistent ability of the lines to induce high rates of reversion and male recombination was successful in determining that the "mutation accumulation lines" do possess mutator factors.


Design of ruthenium-cytochrome c derivatives to measure electron transfer to cytochrome c peroxidase.
A new technique has been introduced to measure interprotein electron transfer which involves photoexcitation of a tris(bipyridine)ruthenium (Ru) complex covalently attached to one of the proteins. Four different strategies have been developed to specifically attach Ru to protein lysine amino groups, histidine imidazole groups, and cysteine sulhydryl groups. These strategies have been used to prepare more than 20 different singly-labeled Ru-cytochrome c derivatives. The new ruthenium photoexcitation technique has been used to study the mechanism for electron transfer between cytochrome c and cytochrome c peroxidase. Laser excitation of a complex between Ru-cytochrome c and cytochrome c peroxidase compound I results in formation of Ru(II*) which is a strong reducing agent, and rapidly transfers an electron to heme c Fe(III) to form Fe(II). The heme c Fe(II) then rapidly transfers an electron to the Trp-191 radical cation in CMPI. The rate constant for this reaction is 6 x 10(4) s-1 for a horse Ru-cytochrome c derivative labeled at lysine 27, and greater than 10(6) s-1 for yeast Ru-cytochrome c derivatives. A second laser flash results in electron transfer from heme c to the oxyferryl heme in cytochrome c peroxidase compound II with a rate constant of 350 s-1. The ruthenium photoreduction technique has been used to study the interaction domain between the two proteins, the pathway for electron transfer to the radical cation and the oxyferryl heme, and the specific residues in the heme crevice which control the electron transfer properties of the Trp-191 radical cation and the oxyferryl heme.


Neural systems for word meaning modulated by semantic ambiguity.
One important issue in neuroimaging research on language is how the brain processes and represents lexical semantics. Past studies with various paradigms reveal that the left inferior prefrontal and mid-superior temporal regions play a crucial role in semantic processing. Those studies, however, typically utilize words having a precise and dominant meaning as stimuli and have not manipulated lexico-semantic ambiguity, a key feature of human language, as an experimental variable. Here, we used a word generation paradigm to examine whether neuroanatomical networks for meaning are modulated by lexical ambiguity. We found that, compared with semantically precise words, semantically ambiguous words were mediated by strong brain activations in the left dorsal-lateral frontal areas, the anterior cingulate, and the right inferior parietal lobe. Semantically precise words, instead, were associated with the left inferior prefrontal and mid-superior temporal sites. These findings indicate that semantic analysis of written words is a dynamic process involving coordination of widely distributed neural subsystems, which are weighted by semantic ambiguity.


Gene therapy for muscular dystrophy: current progress and future prospects.
Muscular dystrophies refer to a group of inherited disorders characterized by progressive muscle weakness, wasting and degeneration. So far, there is no effective treatment but new gene-based therapies are currently being developed with particular noted advances in using conventional gene replacement strategies, RNA-based approaches, or cell-based gene therapy with a main focus on Duchenne muscular dystrophy (DMD). DMD is the most common and severe form of muscular dystrophy and current treatments are far from adequate. However, genetic and cell-based therapies, in particular exon skipping induced by antisense strategies, and corrective gene therapy via functionally engineered dystrophin genes hold great promise, with several clinical trials ongoing. Proof-of-concept of exon skipping has been obtained in animal models, and most recently in clinical trials; this approach represents a promising therapy for a subset of patients. In addition, gene-delivery-based strategies exist both for antisense-induced reading frame restoration, and for highly efficient delivery of functional dystrophin mini- and micro-genes to muscle fibres in vivo and muscle stem cells ex-vivo. In particular, AAV-based vectors show efficient systemic gene delivery to skeletal muscle directly in vivo, and lentivirus-based vectors show promise of combining ex vivo gene modification strategies with cell-mediated therapies.


Social Disadvantage, Access to Care, and Disparities in Physical Functioning Among Children Hospitalized with Respiratory Illness.
Understanding disparities in child health-related quality of life (HRQoL) may reveal opportunities for targeted improvement. This study examined associations between social disadvantage, access to care, and child physical functioning before and after hospitalization for acute respiratory illness. From July 1, 2014, to June 30, 2016, children ages 8-16 years and/or caregivers of children 2 weeks to 16 years admitted to five tertiary care children's hospitals for three common respiratory illnesses completed a survey on admission and within 2 to 8 weeks after discharge. Survey items assessed social disadvantage (minority race/ ethnicity, limited English proficiency, low education, and low income), difficulty/delays accessing care, and baseline and follow-up HRQoL physical functioning using the Pediatric Quality of Life Inventory (PedsQL, range 0-100). We examined associations between these three variables at baseline and follow-up using multivariable, mixed-effects linear regression models with multiple imputation sensitivity analyses for missing data. A total of 1,325 patients and/or their caregivers completed both PedsQL assessments. Adjusted mean baseline PedsQL scores were significantly lower for patients with social disadvantage markers, compared with those of patients with none (78.7 for >3 markers versus 85.5 for no markers, difference -6.1 points (95% CI: -8.7, -3.5). The number of social disadvantage markers was not associated with mean follow-up PedsQL scores. Difficulty/delays accessing care were associated with lower PedsQL scores at both time points, but it was not a significant effect modifier between social disadvantage and PedsQL scores. Having social disadvantage markers or difficulty/delays accessing care was associated with lower baseline physical functioning; however, differences were reduced after hospital discharge.


[Thymic mucosa-associated lymphoid tissue (MALT) lymphoma developing during long-term follow-up for Sjögren's syndrome and idiopathic thrombocytopenic purpura;report of a case].
Mucosa-associated lymphoid tissue (MALT) lymphoma is a rare type of lymphoma that arises in small CD20-positive lymphocytes. We encountered a case of thymic MALT lymphoma treated with surgical intervention during long-term follow-up for Sjögren's syndrome and idiopathic thrombocytopenic purpura (ITP). Although symptomatic remission of Sjögren's syndrome and ITP had already been achieved, the levels of anti-SSA and anti-SSB antibodies remained high. Chronic stimulation by these antibodies may contribute to the development of MALT lymphoma. A careful follow-up may be indicated for this case with a complex immunological background.


The intermediate enzymes of isoprenoid metabolism as anticancer targets.
Inhibitors of isoprenoid biosynthesis are widely used to treat human disease including statins and nitrogenous bisphosphonates. Due to the importance of core human isoprenoid biosynthesis for diverse cellular processes related to cancer cell growth and metastasis, inhibition of this pathway may produce beneficial anticancer consequences. For example, ras oncogenes are well known; ras proteins are overexpressed in many human cancers, and these proteins must be isoprenylated to function. The rho proteins are important for regulating cell motility, and also must be isoprenylated. This has drawn significant attention to inhibitors of protein prenyl transferases. In addition to the reactions that are targeted in current clinical applications, there are other enzymes that have not been studied as extensively. Inhibition of these enzymes, from mevalonate kinase to geranylgeranyl diphosphate synthase, could be attractive as a single agent therapy or in combination with current agents for treatment of cancers in which isoprenylated proteins have been implicated. While detailed in vivo data for many of these putative targets is lacking, there have been several breakthroughs in recent years that could facilitate further studies. In particular, compounds that specifically inhibit some of the downstream isoprenoid biosynthesis enzymes have been developed and their effects in cancer models are emerging. This review will discuss current knowledge of these lesser known isoprenoid pathway enzymes, identify trends in the development of their small molecule inhibitors, and describe the applications and effects of these compounds in cancer models.


Cell wall assembly by Pneumocystis carinii. Evidence for a unique gsc-1 subunit mediating beta -1,3-glucan deposition.
Pneumocystis carinii remains a persistent cause of severe pneumonia in immune compromised patients. Recent studies indicate that P. carinii is a fungal species possessing a glucan-rich cyst wall. Pneumocandin antagonists of beta-1,3-glucan synthesis rapidly suppress infection in animal models of P. carinii pneumonia. We, therefore, sought to define the molecular mechanisms of beta-glucan cell wall assembly by P. carinii. Membrane extracts derived from freshly purified P. carinii incorporate uridine 5'-diphosphoglucose into insoluble carbohydrate, in a manner that was completely inhibited by the pneumocandin L733-560, an antagonist of Gsc-1-type beta-glucan synthetases. Using degenerative polymerase chain reaction and library screening, the P. carinii Gsc-1 catalytic subunit of beta-1,3-glucan synthetase was cloned and characterized. P. carinii gsc1 exhibited homology to phylogenetically related fungal beta-1,3-glucan synthetases, encoding a predicted 214-kDa integral membrane protein with 12 transmembrane domain structure. Immunoprecipitation of P. carinii extracts, with a synthetic peptide anti-Gsc-1 antibody, specifically yielded a protein of 219.4 kDa, which was also capable of incorporating 5'-diphosphoglucose into insoluble glucan carbohydrate. As opposed to other fungi, the expression of gsc-1 mRNA is uniquely regulated over P. carinii's life cycle, having minimal expression in trophic forms, but substantial expression in the thick-walled cystic form of the organism. These results indicate that P. carinii contains a unique catalytic subunit of beta-1,3-glucan synthetase utilized in cyst wall formation. Because synthesis of beta-1,3-glucan is absent in mammalian cells, inhibition of the P. carinii Gsc-1 represents an attractive molecular target for therapeutic exploitation.


S100B-RAGE-mediated augmentation of angiotensin II-induced activation of JAK2 in vascular smooth muscle cells is dependent on PLD2.
Angiotensin II (Ang II), a vasoactive peptide that is also considered a growth factor, has been implicated in both normal and diabetic cellular proliferation. We recently found that activation of janus kinase 2 (JAK2) is essential for the Ang II-induced proliferation of vascular smooth muscle cells (VSMCs) and that high glucose augments Ang II-induced proliferation of VSMCs by increasing signal transduction through activation of JAK2. Here, we demonstrate that S100B, a ligand for the receptor of advanced glycation end products (RAGEs), augmented both Ang II-induced tyrosine phosphorylation of JAK2 and cell proliferation in VSMCs in a receptor-dependent manner. We also found that S100B-RAGE interaction triggered intracellular generation of reactive oxygen species (ROS), VSMC proliferation, and JAK2 tyrosine phosphorylation via activation of phospholipase D (PLD)2. These results provide direct evidence for linkages between PLD2, ROS production, and S100B-RAGE-induced enhancement of Ang II-induced cell proliferation and activation of JAK2 in VSMCs.


Activation of PPAR gamma in colon tumor cell lines by oxidized metabolites of linoleic acid, endogenous ligands for PPAR gamma.
The nuclear hormone receptor peroxisome proliferator-activated receptor (PPAR) gamma plays an important role in the differentiation of intestinal cells and other tissues. Real-time PCR examination of PPAR mRNA for gamma1, gamma2 and gamma3, in Caco-2 and HCT-116 colon cell lines showed that gamma3 is the most abundant message in both lines. Treatment of Caco-2 cells with sodium butyrate, which induces cell differentiation, also leads to an increase in all three PPAR mRNAs. In contrast, treatment of HCT-116 cells with sodium butyrate, which does not lead to differentiation of these cells, causes a decrease in the amount of all three PPAR mRNAs. Furthermore, the amount of PPAR mRNA is greater in Caco-2 cells than in HCT-116 cells at all times examined. As several oxidative metabolites of linoleic acid, including 13-hydroxyoctadecadienoic acid (13-HODE) and 13-oxooctadecadienoic acid (13-OXO) have been shown to bind PPAR, and there is a strong positive correlation between enzymes for metabolism of linoleate oxidation products, intestinal cell differentiation and the distribution of PPAR, we also performed a detailed investigation of the activation of PPAR gamma by 13-HODE and 13-OXO. For these experiments, Caco-2 and HCT-116 cells were transfected with constructs containing PPAR gamma1 or gamma2 then a PPRE-luc reporter construct. Exposure of transfected cells to micromolar concentrations of 13-HODE or 13-OXO produced concentration-dependent increases in luciferase activity. In addition, the two linoleate metabolites activate endogenous PPAR in these cell lines transfected with only PPRE-luc. The data substantiate the contention that oxidation products of linoleic acid are metabolically produced endogenous ligands for PPAR gamma and that PPAR gamma plays an important role in the differentiation of intestinal cells.


Targeting of antitumor ımmune responses with live-attenuated Leishmania strains in breast cancer model.
Cancer is a major cause of death worldwide and most of the therapeutic approaches are relatively ineffective in eliminating cancer especially due to drug resistance. As an alternative, therapy with live microorganisms can induce a robust proinflammatory and anti-cancer immune response in the microenvironment of the tumor. In the present study, we aimed to establish a model for taking the advantages of immune responses against intracellular protozoan parasites for cancer treatment. Leishmania infantum and L. tropica were used in our study as agents of visceral and cutaneous forms of the infection, respectively. After establishing 4T1 breast cancer in mice groups, live-attenuated L. infantum (At-Li) and live-attenuated L. tropica (At-Lt) treatments were performed and results were evaluated according to tumor volume, immune markers and histological examination. Live-attenuated Leishmania strains regressed 4T1-breast cancer in mice and are nonpathogenic, and these strains induce an immune response against 4T1 breast cancer. It is shown that At-Lt is found to be more effective than At-Li in breast cancer treatment using different methods included in the study as analyses of immune parameters, and histopathological examination in tumor tissue besides spleen cells. The tumor grew more slowly by the immune-stimulant effect of live-attenuated Leishmania parasites. This promising therapy should be investigated for optimization in further studies with different cancer types and L. tropica may be designed to express antigens to enhance tumor antigen-specific responses, which may further improve efficacy and immune memory development.


Targeting senescence improves angiogenic potential of adipose-derived mesenchymal stem cells in patients with preeclampsia.
Preeclampsia is a pregnancy-specific hypertensive disorder characterized by impaired angiogenesis. We postulate that senescence of mesenchymal stem cells (MSC), multipotent cells with pro-angiogenic activities, is one of the mechanisms by which systemic inflammation exerts inhibitory effects on angiogenesis in preeclampsia. MSC were isolated from abdominal fat tissue explants removed during medically indicated C-sections from women with preeclampsia (PE-MSC, n = 10) and those with normotensive pregnancies (NP-MSC, n = 12). Sections of the frozen subcutaneous adipose tissue were assessed for inflammation by staining for tumor necrosis factor (TNF)-alpha and monocyte chemoattractant protein (MCP)-1. Viability, proliferation, and migration were compared between PE-MSC vs. NP-MSC. Apoptosis and angiogenesis were assayed before and after treatment with a senolytic agent (1 μM dasatinib) using the IncuCyte S3 Live-Cell Analysis System. Similarly, staining for senescence-associated beta galactosidase (SABG) and qPCR for gene expression of senescence markers, p16 and p21, as well as senescence-associated secretory phenotype (SASP) components, IL-6, IL-8, MCP-1, and PAI-1, were studied before and after treatment with dasatinib and compared between PE and NP. After in vitro exposure to TNF-alpha, MSC demonstrated upregulation of SASP components, including interleukins-6 and -8 and MCP-1. Staining of the subcutaneous adipose tissue sections revealed a greater inflammatory response in preeclampsia, based on the higher levels of both TNF-alpha and MCP-1 compared to normotensive pregnancies (p < 0.001 and 0.024, respectively). MSC isolated from PE demonstrated a lower percentage of live MSC cells (p = 0.012), lower proliferation (p = 0.005), and higher migration (p = 0.023). At baseline, PE-MSC demonstrated a senescent phenotype, reflected by more abundant staining for SABG (p < 0.001), upregulation of senescence markers and SASP components, as well as lower angiogenic potential (p < 0.001), compared to NP-MSC. Treatment with dasatinib increased significantly the number of apoptotic PE-MSC compared to NP-MSC (0.011 vs. 0.093) and decreased the gene expression of p16 and six SASP components. The mechanistic link between senescence and impaired angiogenesis in PE was confirmed by improved angiogenic potential of PE-MSC (p < 0.001) after dasatinib treatment. Our data suggest that MSC senescence exerts inhibitory effects on angiogenesis in preeclampsia. Senolytic agents may offer the opportunity for mechanism-based therapies.


Lifestyle Modifications and Policy Implications for Primary and Secondary Cancer Prevention: Diet, Exercise, Sun Safety, and Alcohol Reduction.
Improved cancer treatments and cancer detection methods are not likely to completely eradicate the burden of cancer. Primary prevention of cancer is a logical strategy to use to control cancer while also seeking novel treatments and earlier detection. Lifestyle modification strategies to improve primary prevention and risk reduction for the development of cancer include choosing a healthy diet with an emphasis on plant sources, maintaining a healthy weight throughout life, being physically active, regularly using sunscreen and wearing protective clothing, limiting sun exposure during the hours of 10 AM to 2 PM, avoiding indoor tanning, and reducing or eliminating alcohol use. In addition to continued use of ongoing education of the public, health care providers, and cancer support communities, other policy and public health efforts should be pursued as well. Examples of supported and successful policy approaches are included in this article, including efforts to limit indoor tanning and improve community-wide interventions to reduce ultraviolet radiation exposure as well as to formally support various alcohol policy strategies including increasing alcohol taxes, reducing alcohol outlet density, improving clinical screening for alcohol use disorders, and limiting youth exposure to alcohol marketing and advertising. These prevention strategies are expected to have the largest impact on the development of melanoma as well as breast, colorectal, head and neck, liver, and esophageal cancers. The impact of these strategies as secondary prevention is less well understood. Areas of additional needed research and implementation are also highlighted. Future areas of needed research are the effects of these modifications after the diagnosis of cancer (as secondary prevention).


Alzheimer's Disease-Related Neuropathology Among Patients with Medication Treated Type 2 Diabetes in a Community-Based Autopsy Cohort.
Diabetes is a risk factor for Alzheimer's disease and related dementias (ADRD). Epidemiologic evidence shows an association between diabetes medications and ADRD risk; cell and mouse models show diabetes medication association with AD-related neuropathologic change (ADNC). This hypothesis-generating analysis aimed to describe autopsy-measured ADNC for individuals who used diabetes medications. Descriptive analysis of ADNC for Adult Changes in Thought (ACT) Study autopsy cohort who used diabetes medications, including sulfonylureas, insulin, and biguanides; total N = 118. ADNC included amyloid plaque distribution (Thal phasing), neurofibrillary tangle (NFT) distribution (Braak stage), and cortical neuritic plaque density (CERAD score). We also examined quantitative measures of ADNC using the means of standardized Histelide measures of cortical PHF-tau and Aβ1-42. Adjusted analyses control for age at death, sex, education, APOE genotype, and diabetes complication severity index. Adjusted analyses showed no significant association between any drug class and traditional neuropathologic measures compared to nonusers of that class. In adjusted Histelide analyses, any insulin use was associated with lower mean levels of Aβ1-42 (-0.57 (CI: -1.12, -0.02)) compared to nonusers. Five years of sulfonylureas and of biguanides use was associated with lower levels of Aβ1-42 compared to nonusers (-0.15 (CI: -0.28, -0.02), -0.31 (CI: -0.54, -0.07), respectively). Some evidence exists that diabetes medications are associated with lower levels of Aβ1-42, but not traditional measures of neuropathology. Future studies are needed in larger samples to build understanding of the mechanisms between diabetes, its medications, and ADRD, and to potentially repurpose existing medications for prevention or delay of ADRD.


Common Sequence Variation in the VEGFC Gene Is Associated with Diabetic Retinopathy and Diabetic Macular Edema.
To investigate associations between single nucleotide polymorphisms (SNPs) in the VEGFC gene and the development of diabetic retinopathy (DR) in white patients with type 1 diabetes mellitus (T1DM) or type 2 diabetes mellitus (T2DM). Cross-sectional, case control study. White patients with T1DM or T2DM (n = 2899) were recruited from ophthalmology and endocrine clinics in Australia and the United Kingdom. Patients with T2DM were required to have diabetes mellitus (DM) for at least 5 years and be receiving oral hypoglycemic treatment or insulin. Participants were categorized according to their worst-ever DR grading, as having "no DR" (no history of nonproliferative DR [NPDR], proliferative DR [PDR], or diabetic macular edema [DME]) or "any DR" (further subclassified as NPDR or PDR, without or with DME). Clinical characteristics, glycemic control (hemoglobin A1c [HbA1c]), and presence of diabetic complications were determined at recruitment. Genotyping was performed for 13 VEGFC tag SNPs. Odds ratios (ORs) were determined for associations with DR of VEGFC tag SNPs, individually and within haplotypes. Logistic regression was used to adjust for clinical covariates, including DM type, age, sex, DM duration, hypertension, nephropathy, HbA1c, and smoking. Participants with DM but "no DR" (n = 980) were compared with 1919 participants with DM and "any DR." Three VEGFC SNPs were associated with DR after logistic regression: rs17697419 (P = 0.001; OR, 0.67; confidence interval [CI], 0.52-0.85), rs17697515 (P = 0.001; OR, 0.62; CI, 0.47-0.81), and rs2333526 (P = 0.005; OR, 0.69; CI, 0.54-0.90). rs17697515 Was also specifically associated with DME in those with T2DM (P = 0.004; OR, 0.53; CI, 0.35-0.82). Haplotype analysis revealed 2 significantly associated haplotypes, both protective against DR development. Significant associations were found between VEGFC tag SNPs (individually and within haplotypes) and the presence of any DR or DME in white participants with T1DM and T2DM.


PET/CT with Fluorodeoxyglucose During Neoadjuvant Chemoradiotherapy in Locally Advanced Rectal Cancer.
The aim of the present study is to evaluate the accuracy of Positron Emission Tomography/Computed Tomography (PET/CT) with Fluorodeoxyglucose ([18F]FDG) to predict treatment response in patients with locally advanced rectal cancer (LARC) during neoadjuvant chemoradiotherapy. Forty-one LARC patients performed [18F]FDG-PET/CT at baseline (PET0). All patients received continuous capecitabine concomitant to radiotherapy on the pelvis, followed by intermittent capecitabine until two weeks before curative surgery. [18F]FDG-PET/CT was also carried out at 40 Gy-time (PET1) and at the end of neoadjuvant therapy (PET2). PET imaging was analysed semi-quantitatively through the measurement of maximal standardised uptake value (SUVmax) and the tumour volume (TV). Histology was expressed through pTNM and Dworak tumor regression grading. Patients were categorised into responder (downstaging or downsizing) and non-responder (stable or progressive disease by comparison pretreatment parameters with clinical/pathological characteristics posttreatment/after surgery). Logistic regression was used to evaluate SUVmax and TV absolute and percent reduction as predictors of response rate using gender, age, and CEA as covariates. Progression-free survival (PFS) and overall survival (OS) were estimated by the Kaplan-Meier method. Survivals were compared by the Log-Rank test. Twenty-three responders (9 ypCR, 14 with downstaged disease) and 18 non-responders showed differences in terms of both early and posttreatment SUVmax percent reduction (median comparison: responder = 63.2%, non-responder = 44.2%, p = 0.04 and responder = 76.9%, non-responder = 61.6%, p = 0.06 respectively). The best predictive cut-offs of treatment response for early and posttreatment SUVmax percent reduction were ≥57% and ≥66% from baseline (p = 0.02 and p = 0.01 respectively). [18F]FDG-PET/CT is a reliable technique for evaluating therapy response during neoadjuvant treatment in LARC, through a categorical classification of the SUV max reduction during treatment.


Self-reported neurological symptoms in relation to CO emissions due to problem gas appliance installations in London: a cross-sectional survey.
Previous research by the authors found evidence that up to 10% of particular household categories may be exposed to elevated carbon monoxide (CO) concentrations from poor quality gas appliance installations. The literature suggests certain neurological symptoms are linked to exposure to low levels of CO. This paper addresses the hypothesis that certain self-reported neurological symptoms experienced by a householder are linked to an estimate of their CO exposure. Between 27 April and 27 June 2006, 597 homes with a mains supply of natural gas were surveyed, mainly in old, urban areas of London. Qualified gas engineers tested all gas appliances (cooker, boiler, gas fire, and water heater) and reported, according to the Gas Industry Unsafe Situations Procedure, appliances considered At Risk (AR), Immediately Dangerous (ID) or Not to Current Standards (NCS). Five exposure risk categories were defined based on measurement of CO emitted by the appliance, its features and its use, with "high or very high" exposure category where occupants were considered likely to be exposed to levels greater than 26 ppm for one hour. The prevalence of symptoms at each level of exposure was compared with that at lowest level of exposure. Of the households, 6% were assessed as having a "high or very high" risk of exposure to CO. Of the individuals, 9% reported at least one neurological symptom. There was a statistically significant association between "high or very high" exposure risk to CO and self-reported symptoms compared to "no exposure" likelihood, for households not in receipt of benefit, controlling for "number of residents" and presence of pensioners, OR = 3.23 (95%CI: 1.28, 8.15). Risk ratios across all categories of exposure likelihood indicate a dose-response pattern. Those households in receipt of benefit showed no dose-response pattern. This study found an association between risk of CO exposure at low concentration, and prevalence of self-reported neurological symptoms in the community for those households not in receipt of benefit. As health status was self-reported, this association requires further investigation.


Delineating the mode of action of adenosine A1 receptor allosteric modulators.
Despite the identification of 2-amino-3-benzoylthiophenes (2A3BTs) as the first example of small-molecule allosteric potentiators of agonist function at a G protein-coupled receptor (GPCR)-the adenosine A(1) receptor-their mechanism of action is still not fully understood. We now report the mechanistic basis for the complex behaviors noted for 2A3BTs at A(1) receptors. Using a combination of membrane-based and intact-cell radioligand binding, multiple signaling assays, and a native tissue bioassay, we found that the allosteric interaction between 2A3BTs and the agonists 2-chloro-N(6)-[(3)H]cyclopentyladenosine or (-)-N(6)-(2-phenylisopropyl)adenosine (R-PIA) or the antagonist [(3)H]8-cyclopentyl-1,3-dipropylxanthine is consistent with a ternary complex model involving recognition of a single extracellular allosteric site. However, when allowed access to the intracellular milieu, 2A3BTs have a secondary action as direct G protein inhibitors; this latter property is receptor-independent as it is observed in nontransfected cells and also after stimulation of another GPCR. In addition, we found that 2A3BTs can signal as allosteric agonists in their own right but show bias toward certain pathways relative to the orthosteric agonist, R-PIA. These results indicate that 2A3BTs have a dual mode of action when interacting with the A(1) receptor and that they can engender novel functional selectivity in A(1) signaling. These mechanisms need to be factored into allosteric ligand structure-activity studies.


Compact bis-adduct fullerenes and additive-assisted morphological optimization for efficient organic photovoltaics.
Bis-adduct fullerenes surrounded by two insulating addends sterically attenuate intermolecular interaction and cause inferior electron transportation. In this research, we have designed and synthesized a new class of bis-adduct fullerene materials, methylphenylmethano-C60 bis-adduct (MPC60BA), methylthienylmethano-C60 bis-adduct (MTC60BA), methylphenylmethano-C70 bis-adduct (MPC70BA), and methylthienylmethano-C70 bis-adduct (MTC70BA), functionalized with two compact phenylmethylmethano and thienylmethylmethano addends via cyclopropyl linkages. These materials with much higher-lying lowest unoccupied molecular orbital (LUMO) energy levels successfully enhanced the Voc values of the P3HT-based solar cell devices. The compact phenylmethylmethano and thienylmethylmethano addends to promote fullerene intermolecular interactions result in aggregation-induced phase separation as observed by the atomic force microscopy (AFM) and transmission electron microscopy (TEM) images of the poly(3-hexylthiophene-2,5-diyl) (P3HT)/bis-adduct fullerene thin films. The device based on the P3HT/MTC60BA blend yielded a Voc of 0.72 V, a Jsc of 5.87 mA/cm(2), and a fill factor (FF) of 65.3%, resulting in a power conversion efficiency (PCE) of 2.76%. The unfavorable morphologies can be optimized by introducing a solvent additive to fine-tune the intermolecular interactions. 1-Chloronaphthalene (CN) having better ability to dissolve the bis-adduct fullerenes can homogeneously disperse the fullerene materials into the P3HT matrix. Consequently, the aggregated fullerene domains can be alleviated to reach a favorable morphology. With the assistance of CN additive, the P3HT/MTC60BA-based device exhibited enhanced characteristics (a Voc of 0.78 V, a Jsc of 9.04 mA/cm(2), and an FF of 69.8%), yielding a much higher PCE of 4.92%. More importantly, the additive-assisted morphological optimization is consistently effective to all four compact bis-adduct fullerenes regardless of the methylphenylmethano or methylthienylmethano scaffolds as well as C60 or C70 core structures. Through the extrinsic additive treatment, these bis-adduct fullerene materials with compact architectures show promise for high-performance polymer solar cells.


Depurination of DNA as a possible mutagenic pathway for cells.
The possible consequences of depurination for both spontaneous and induced mutagenesis were investigated using in vitro and in vivo assays. Depurination of synthetic polynucleotide templates such as poly [d(A-T)] or poly [d(G-C)] leads to increased misincorporation of noncomplementary nucleotides when these templates are copied by prokaryotic and eukaryotic DNA polymerases. The ability of Escherichia coli DNA polymerase I to copy over apurinic sites was demonstrated using single-stranded circular DNA of bacteriophage 0X174 as a template and starting DNA synthesis at a fixed point. Analysis of the newly synthesized 0X174 restriction fragments on neutral and alkaline sucrose gradients shows that synthesis proceeded past apurinic sites. When using depurinated 0X174 DNA containing the am3 amber mutation as a template for copying by E. coli DNA polymerase I, an increased reversion to wild type is observed after transfection into E. coli spheroplasts. The enhancement in reversion frequency is proportional to the extent of depurination, suggesting that depurination is also mutagenic during copying natural DNA in vitro. When noncopied depurinated 0X174 am3 DNA is transfected in E. coli spheroplasts, no increase in reversion frequency is observed above background level. However, when the spheroplasts are derived from bacteria in which the SOS response had been induced by UV irradiation, a substantial increase is observed for depurinated molecules, whereas no increase is observed for nondepurinated templates, suggesting in vivo mutagenesis at depurinated sites. In each of the different assay systems investigated, the increase in misincorporation or reversion frequency is a linear function of the number of sites and is abolished by treatment of the depurinated templates with alkali, which rapidly induces strand breakage at apurinic sites.


Characterization of Halomonas sp. ZM3 isolated from the Zelazny Most post-flotation waste reservoir, with a special focus on its mobile DNA.
Halomonas sp. ZM3 was isolated from Zelazny Most post-flotation mineral waste repository (Poland), which is highly contaminated with heavy metals and various organic compounds. Mobile DNA of the strain (i.e. plasmids and transposons) were analyzed in order to identify genetic information enabling adaptation of the bacterium to the harsh environmental conditions. The analysis revealed that ZM3 carries plasmid pZM3H1 (31,370 bp), whose replication system may be considered as an archetype of a novel subgroup of IncU-like replicons. pZM3H1 is a narrow host range, mobilizable plasmid (encodes a relaxase of the MOBV family) containing mercury resistance operon (mer) and czcD genes (mediate resistance to zinc and cobalt), which are part of a large truncated Tn3 family transposon. Further analysis demonstrated that the phenotypes determined by the pZM3H1 resistance cassette are highly dependent on the host strain. In another strand of the study, the trap plasmid pMAT1 was employed to identify functional transposable elements of Halomonas sp. ZM3. Using the sacB positive selection strategy two insertion sequences were identified: ISHsp1 - representing IS5 group of IS5 family and ISHsp2 - a distinct member of the IS630 family. This study provides the first detailed description of mobile DNA in a member of the family Halomonadaceae. The identified IncU plasmid pZM3H1 confers resistance phenotypes enabling adaptation of the host strain to the Zelazny Most environment. The extended comparative analysis has shed light on the distribution of related IncU plasmids among bacteria, which, in many cases, reflects the frequency and direction of horizontal gene transfer events. Our results also identify plasmid-encoded modules, which may form the basis of novel shuttle vectors, specific for this group of halophilic bacteria.


The contralateral transcingulate approach: operative technique and results with vascular lesions.
The contralateral transcingulate approach is a variation of the contralateral transcallosal approach for lesions located in the cingulate gyrus and deep white matter of the medial frontal lobe. To more thoroughly describe the contralateral transcingulate approach by reporting our operative technique and results. This approach positions the head with the midline horizontally, lesion on the upside, allowing gravity retraction of the dependent frontal lobe. Bifrontal craniotomy and splitting of the interhemispheric fissure create a crossing trajectory from the contralateral fissure to the ipsilateral cingulate gyrus that maximizes lateral exposure. Eleven patients with vascular lesions were treated with the contralateral transcingulate approach (9 patients with cavernous malformations and 2 patients with arteriovenous malformations). Eight lesions were located on the left side, 5 in the cingulate gyrus, and 6 in the deep frontal lobe. The falx was cut in 5 patients to extend the crossing trajectory. All lesions were removed completely, with neurological morbidity in 1 patient caused by venous infarction. Although similar to the contralateral transcallosal approach, the contralateral transcingulate approach accesses lesions outside the ventricle and has a steeper crossing trajectory. This approach requires no disruption of brain tissue with lesions on the cingulate surface and only a small incision in cingulate gyrus with lesions in the deep frontal lobe. The ipsilateral pericallosal and callosomarginal arteries provide dependable landmarks for transcingulate dissection. The contralateral transcingulate approach offers an alternative medial approach to lesions near language and motor areas and avoids lateral transcortical approaches, awake speech mapping, and risk to eloquent cortex in the dominant hemisphere.


Polymer-ionic liquid ternary systems for Li-battery electrolytes: Molecular dynamics studies of LiTFSI in a EMIm-TFSI and PEO blend.
This paper presents atomistic molecular dynamics simulation studies of lithium bis(trifluoromethane)sulfonylimide (LiTFSI) in a blend of 1-ethyl-3-methylimidazolium (EMIm)-TFSI and poly(ethylene oxide) (PEO), which is a promising electrolyte material for Li- and Li-ion batteries. Simulations of 100 ns were performed for temperatures between 303 K and 423 K, for a Li:ether oxygen ratio of 1:16, and for PEO chains with 26 EO repeating units. Li(+) coordination and transportation were studied in the ternary electrolyte system, i.e., PEO16LiTFSI⋅1.0 EMImTFSI, by applying three different force field models and are here compared to relevant simulation and experimental data. The force fields generated significantly different results, where a scaled charge model displayed the most reasonable comparisons with previous work and overall consistency. It is generally seen that the Li cations are primarily coordinated to polymer chains and less coupled to TFSI anion. The addition of EMImTFSI in the electrolyte system enhances Li diffusion, associated to the enhanced TFSI dynamics observed when increasing the overall TFSI anion concentration in the polymer matrix.


Perspectives of Cone-beam Computed Tomography in Interventional Radiology: Techniques for Planning, Guidance, and Monitoring.
Cone-beam computed tomography (CBCT) has emerged as a prominent imaging modality in interventional radiology that offers real-time visualization and precise guidance in various procedures. This article aims to provide an overview of the techniques used to guide and monitor interventions that use CBCT. It discusses the advantages of CBCT, its current applications, and potential future CBCT-related developments in the field of interventional radiology.


Who treats medical conditions more cost efficiently?
Total costs in HMOs are 10% to 40% below those in indemnity plans; however, investigators have not examined the types of medical conditions that HMO providers treat more cost efficiently than indemnity providers. The present study was designed to evaluate if managed care health plan specialists treat complex conditions as cost efficiently as the HMO primary care providers treated the less-complex conditions and, if not, why?


A Single-Arm, Open-Label, Pilot, and Feasibility Study of a High Nicotine Strength E-Cigarette Intervention for Smoking Cessation or Reduction for People With Schizophrenia Spectrum Disorders Who Smoke Cigarettes.
An estimated 60%-90% of people with schizophrenia smoke, compared with 15%-24% of the general population, exacerbating the already high morbidity and mortality rates observed in this population. This study aimed to assess the feasibility of using a new-generation high strength nicotine e-cigarette to modify smoking behavior in individuals with schizophrenia spectrum disorders who smoke cigarettes. A single-arm pilot study was conducted with 40 adults with schizophrenia spectrum disorders who smoked and did not intend to reduce or quit smoking. Participants were given a 12-week supply of a JUUL e-cigarette loaded with a 5% nicotine pod. The primary outcome was smoking cessation at week 12. Additional outcomes included: smoking reduction, continuous abstinence at week 24, adoption rate, adherence to the e-cigarette, feasibility, acceptability, and subjective effects. Sixteen (40%) participants quit by the end of 12 weeks. For the whole sample, we observed an overall, sustained 50% reduction in smoking or smoking abstinence in 37/40 (92.5%) of participants and an overall 75% reduction in median cigarettes per day from 25 to six was observed by the end of the 12 weeks (p < .001). A high strength nicotine e-cigarette has the potential to help people with schizophrenia spectrum disorders to quit or reduce smoking. Further research with a larger sample and a comparator group is needed. The results provide useful information and direction to augment the existing body of knowledge on smoking cessation for people with schizophrenia spectrum disorders. Considering that most people with schizophrenia spectrum disorders continue smoking, alternative and efficient interventions to reduce or prevent morbidity and mortality are urgently needed. This study showed that adults who smoke and were not motivated to quit, when provided a new-generation e-cigarette with high nicotine content, demonstrated substantially decreased cigarette consumption without causing significant side effects. Although not specifically measured in this study, nicotine absorption in new-generation devices has been shown to be consistently superior compared with the first generation of e-cigarette devices, and this may help explain the lower quit rates in studies using earlier generation devices.


Osteoporosis is associated with antiepileptic drugs: a population-based study.
Controversy remains regarding the risk of bone abnormalities due to enzyme-inducing antiepileptic drugs (EIAEDs) and non-enzyme-inducing antiepileptic drugs (NEIAEDs). This case-control study aimed to investigate the possible association between osteoporosis and epilepsy disease and AEDs therapy using a population-based dataset in Taiwan. We first identified 48,102 cases, ≥ 18 years of age, who received a first-time diagnosis of osteoporosis, and then randomly selected 144,306 controls. We used conditional logistic regression analyses to compute the odds ratio (OR) and corresponding 95% confidence interval (CI) to compare a previous diagnosis of epilepsy between cases and controls. We found that of the 192,408 sampled subjects, epilepsy was found in 117 (0.24%) cases and 240 (0.17%) controls (p<0.001). Cases were found to be more likely to have previously been diagnosed with epilepsy than controls (OR: 1.41, 95% CI: 1.11 ≈ 1.78, p<0.01), after taking confounders into consideration. Furthermore, we found that, compared to controls, the adjusted OR of cases in which enzyme-inducing AEDs had been prescribed was 2.06 (95% CI: 1.43 ≈ 2.95). A higher proportion of cases with prescribed NEIAED was also found (OR: 2.09, 95% CI: 1.49 ≈ 2.92) compared to controls. This study demonstrates that patients with osteoporosis were more likely to have epilepsy and receive EIAED or NEIAED treatment. For patients with epilepsy who take AEDs, attention should be paid to the adverse effects of osteoporosis.


Identification of three internalization sequences in the cytoplasmic tail of the 46 kDa mannose 6-phosphate receptor.
The cytoplasmic tail of the human 46 kDa mannose 6-phosphate receptor (MPR 46) is necessary for rapid internalization of the receptor and sufficient to mediate internalization of a resident plasma membrane protein. To localize the internalization sequences within the 67 amino acids of the cytoplasmic tail, the tail was progressively shortened from its C-terminus, internal deletions of between four and eight amino acids were introduced into the tail, and individual residues were substituted by alanine, glycine or serine. Three sequences were identified that contribute to the internalization of MPR 46. The first is located within the 23 juxtamembrane cytoplasmic residues of the tail. It contains four essential residues within a heptapeptide and does not resemble known internalization signals. The second sequence contains as a critical residue Tyr-45. The third region is located within the C-terminal seven residues and contains a di-leucine pair as essential residues. The first and third sequences were shown to function as autonomous internalization sequences. Substitution of critically important residues within a single internalization sequence was tolerated, with no or only a moderate decrease in the internalization rate. When essential residues from two or all three internalization sequences were substituted, however, the internalization rate was decreased by more than 60% and 90% respectively. This indicates that the autonomous internalization signals in the cytoplasmic tail of MPR 46 function in an additive manner, but are partly redundant.


Cyberbullying Prevention for Adolescents: Iterative Qualitative Methods for Mobile Intervention Design.
Cybervictimization among adolescents is associated with multiple negative mental health consequences. Although pediatricians often screen for cyberbullying, validated and acceptable programs to reduce the frequency and impact of adolescent cybervictimization are lacking. This study uses agile qualitative methods to refine and evaluate the acceptability of a mixed-modality intervention, initiated within the context of usual pediatric care, for adolescents with a history of cyberharassment and cyberbullying victimization. Three groups of adolescents were successively recruited from an urban primary care clinic to participate in three consecutive iterations (1, 2, and 3) of the program, which consisted of a brief in-clinic intervention followed by 8 weeks of daily, automated SMS text messaging. After 2 weeks of messaging, iteration 1 (I1) participants completed semistructured interviews regarding intervention experiences. Participant feedback was evaluated via framework matrix analysis to guide changes to the program for iteration 2 (I2). Feedback from 2-week interviews of I2 participants was similarly used to improve the program before initiating iteration 3 (I3). Participants in all 3 iterations completed the interviews after completing the program (8 weeks). Daily response rates assessed participant engagement, and satisfaction questionnaires assessed acceptability. A total of 19 adolescents (aged 13-17 years) reporting past-year cybervictimization were enrolled: 7 in I1, 4 in I2, and 8 in I3. Demographic variables included the following: a mean age of 15 (SD 1.5) years; 58% (11/19) female, 42% (8/19) male, 63% (12/19) Hispanic, 37% (7/19) non-Hispanic, 79% (15/19) people of color, and 21% (4/19) White. A total of 73% (14/19) self-identified as having a low socioeconomic status, and 37% (7/19) self-identified as lesbian, gay, or bisexual. The average past 12-month cybervictimization score at baseline was 8.2 (SD 6.58; range 2-26). Participant feedback was used to iteratively refine intervention content and design. For example, participants in I1 recommended that the scope of the intervention be expanded to include web-based conflicts and drama, rather than narrowly focusing on cyberbullying prevention. On the basis of this feedback, the I2 content was shifted toward more general de-escalation skills and bystander empowerment. Overall, 88.34% (940/1064) of the daily queries sent to participants across all 3 iterations received a reply. Participant satisfaction improved considerably with each iteration; 0% (0/7) of I1 participants rated the overall quality of Intervention to Prevent Adolescent Cybervictimization with Text message as excellent, compared to 50% (2/4) of I2 participants and 86% (6/7) of I3 participants. Engagement also improved between the first and third iterations, with participants replying to 59.9% (235/392) of messages in I1, compared to 79.9% (358/488) of messages in I3. This study shows the value of structured participant feedback gathered in an agile intervention refinement methodology for the development of a technology-based intervention targeting adolescents.


Block of outward current in cardiac Purkinje fibers by injection of quaternary ammonium ions.
We have studied the effects of iontophoretic injection of the quaternary ammonium compounds tetraethylammonium (TEA) and tetrabutylammonium (TBA) in cardiac purkinje fibers. We find that TBA(+) is a more effective blocker than TEA(+), but injection of either compound reduces the time-dependent outward plateau currents, transient outward current (I(to)), and the delayed rectifier (I(x)). Our findings provide evidence that these outward cardiac currents are carried by channels that in some respects are pharmacologically similar to squid axon potassium channels. We demonstrate that this procedure is a new tool that can be useful in the analysis of membrane currents in the heart.


Transient pituitary ACTH-dependent Cushing syndrome caused by an immune checkpoint inhibitor combination.
Immune checkpoint inhibitors have improved survival in numerous advanced malignancies, but are associated with a number of immune-related adverse events, including endocrinopathies. Endogenous Cushing's syndrome (CS) is a rare disorder resulting from exposure to high levels of circulating cortisol. CS can be caused either by adrenal cortex tumors or hyperplasia or by pituitary or extra-pituitary tumors over-secreting ACTH (known as ACTH-dependent CS). We report the first case of transient ACTH-dependent CS, which appeared after combined ipilimumab and nivolumab therapy. Our patient presented typical clinical features of CS after three infusions of combined therapy, high serum and daily urinary free cortisol, and high serum ACTH levels. Pituitary MRI showed an enlargement of the pituitary gland suggesting ACTH secretion of pituitary origin, which was confirmed by inferior petrosal sinus sampling. The pituitary findings were preceded by thyroiditis. The evolution was characterized by spontaneous CS regression and subsequent appearance of severe corticotroph deficiency consistent with destructive hypophysitis. Immunotherapy is a novel cause of CS.


Pharyngocutaneous fistulae after total laryngectomy: a systematic review.
Nursing care to patients after total laryngectomy should be based on scientific knowledge. Evidence-based practice is a type of approach that stimulates the use of research results in clinical practice. This study presents a systematic review that aimed to identify the main treatments for pharyngocutaneous fistulae after total laryngectomy. Articles were selected from five databases: Pubmed, Cinahl, Biomednet Research Tools, Cochrane Library and Lilacs. The review sample consisted of 37 articles. After analyzing the articles included in the review, results showed that conservative treatment is commonly used for pharyngocutaneous fistulae, recommending intensive hygiene and wound treatment. The use of research results in clinical practice will grant greater consistency to nurses' actions in care for patients with pharyngocutaneous fistulae.


Effects of antenatal steroid therapy on neurodevelopment in an IUGR mouse model.
To investigate the neurodevelopmental response in postnatal mice secondary to antenatal steroid treatment in association with maternal protein restriction. C57BL/6N pregnant mice (n = 24; 4 per treatment group) were administered control (C) or protein-restricted (PR) diets and subjected to daily subcutaneous injection stress during late gestation (E10-E17) with either 100 microl/kg of dexamethasone sodium phosphate in normosaline (C-D/S, PR-D/S) or normosaline alone (C-S, PR-S). Non-treatment groups were also included (C, PR). Brain samples of pups were collected on postnatal day 7 and analyzed by immunohistochemistry and qRT-PCR. Neonatal weights in the treatment groups were smaller than their counterparts in the C group, but there were no significant differences in brain size. Immunohistochemical evaluation of neuroglial cells revealed a pronounced effect of protein restriction on oligodendrocytes and oligodendrocyte precursor cells with distinct fetal responses to stress and dexamethasone. Further evaluation using quantitative RNA analysis showed significant activation of Galr1, Galr2, Igfbp-1, Igfbp-3, Igfbp-6, and Fgf2 by 1- to 2.5-fold in the PR-D/S group and by much higher increments, 1- to 10.5-fold, in the PR-S group. This preliminary investigation revealed the possible role of dexamethasone in further increasing vulnerability to cell damage in injury-prone neuroglial cells. The distribution of key glial markers and the overexpression of several neurotrophic factors depicted ongoing cellular adaptation.


Predicting the quality of enhanced wideband speech with a cochlear model.
Objective measures are commonly used in the development of speech coding algorithms as an adjunct to human subjective evaluation. Predictors of speech quality based on models of physiological or perceptual processing tend to perform better than measures based on simple acoustical properties. Here, a modeling method based on a detailed physiological model and a neurogram similarity measure is developed and optimized to predict the quality of an enhanced wideband speech dataset. A model capturing temporal modulations in neural activity up to 267 Hz was found to perform as well as or better than several existing objective quality measures.


Botox for the brain: enhancement of cognition, mood and pro-social behavior and blunting of unwanted memories.
It has been suggested that the recent rapid developments in the fields of neuroscience and psychopharmacology have increased the possibilities for pharmacological enhancement of mental functioning. Here, evidence is reviewed which shows that drugs acting on a variety of neurotransmitter systems can indeed enhance cognition, and to a lesser extent mood and pro-social behavior. Moreover, it seems possible to interfere with the (re)consolidation of traumatic memories. There are, however, a number of caveats: first, as cognition-enhancing drugs can simultaneously exert both linear and quadratic (U-shaped) effects, doses most effective in facilitating one behavior could at the same time exert null or even detrimental effects on other cognitive domains. Second, individuals with a 'low memory span' might benefit from cognition-enhancing drugs, whereas 'high span subjects' are 'overdosed'. And finally, evidence suggests that a number of trade-offs could occur. For example, increases of cognitive stability might come at the cost of a decreased capacity to flexibly alter behavior. A short overview of ethical issues raised by the use of cognition and mood enhancing drugs demonstrates the tremendous variety in views and opinions regarding the subject.


Acute mesenteric venous thrombosis in a pregnant woman at 35 weeks of gestation: a case report and review of the literature.
Mesenteric venous thrombosis (MVT) is an infrequent thrombotic event that can cause devastating intestinal hemorrhagic ischemia. The mortality rate among patients with acute MVT ranges from 20 to 50%. Occurrence of MVT in pregnancy is quite rare. In this case report, we describe a pregnant woman who presented with acute MVT at 35 weeks of gestation. Our case was a 26-year-old primigravid woman at 35 weeks gestation. She presented to Guangzhou First People's Hospital with complaints of abdominal pain, nausea and vomiting. The second day after admission, she complained of more intense abdominal pain, anorexia, vomiting and abdominal distention that were out of proportion to physical signs. An emergency exploratory laparotomy was performed. The entire ileum, part of the jejunum and part of the ascending colon were gangrenous, and thromboembolism was discovered in the corresponding mesenteric veins. The necrotic intestine was resected and an end-to-end jejunum-colon anastomosis was performed. A cesarean section was performed to remove the placenta and fetus, which had expired. Histopathological analysis revealed extensive edema, hemorrhage, inflammatory infiltration and necrosis in the resected bowel, and widespread thrombosis in mesenteric venous lumens. The diagnosis of MVT during pregnancy is very difficult due to its low incidence, and non-characteristic symptoms, signs and laboratory results. MVT may be the underlying cause of severe abdominal pain during pregnancy and should be included in the differential diagnosis of pregnant patients with an acute abdomen.


Preclinical and clinical development of novel agents that target the protein kinase C family.
Protein kinase C (PKC) family comprises more than 12 serine-/threonine-specific isoenzymes. PKC isoenzymes play a complex role in the transduction of signal from tyrosine kinase receptor modulating proliferation, cell cycle, apoptosis, invasion, differentiation, and senescence in both cancer cells and endothelial cells. Thereby, inhibition and/or activation of specific PKCs is thought to control tumor growth by interacting directly with cancer cells and indirectly by blocking tumor angiogenesis. Furthermore, PKCs are known to modulate multi-drug resistance, providing a rational for the combination of PKCs modulators with classical cytotoxic drugs. During the past few years, preclinical and clinical data with first-generation PKC inhibitors/activators provided insight that PKCs may indeed represent attractive targets for the discovery of small molecules with new anticancer properties. In this review, we will provide an overview on the current understanding of PKC participation in chemotherapeutic resistance, the possible implications in cancer treatment, and the potential of most recent PKC inhibitors in molecular cancer therapeutics.


Contextual effects on force control and timing in a finger-tapping sequence with an accentuated- or attenuated-force tap.
An experiment was conducted to examine contextual effects of the magnitude of changes in force on force control in a finger-tapping sequence with an accentuated- (accentuated-force condition) or attenuated-force tap (attenuated-force condition). Participants were trained to produce a finger-tapping sequence with an intertap interval of 500 ms and four force patterns. During practice, visual force feedback pertaining to the two target forces in the tapping sequences was provided. After practice, the participants reproduced the learned tapping sequences in the absence of feedback. A main result was that the last accentuated-force tap affected the first three taps of the tapping sequence. For the accentuated-force conditions, the larger the difference between the first three target forces and the last target force, the larger the first three forces. This indicates the contextual effect of serial position for force control. This effect was not observed, however, under the attenuated-force conditions.


Translational regulation of HIV-1 replication by HIV-1 Rev cellular cofactors Sam68, eIF5A, hRIP, and DDX3.
Nuclear export and translation of HIV-1 RNA are two important posttranscriptional events for HIV-1 gene expression and replication. HIV-1 Rev functions to export unspliced and incompletely spliced HIV-1 RNA from the nucleus to the cytoplasm; it requires interaction with several cellular cofactors such as Sam68, eIF5A, hRIP, and DDX3. Meanwhile, some studies have also implicated Rev and some of its cofactors such as Sam68 in HIV-1 RNA translation. Thus, in this study, we aimed to characterize the potential function of all these four Rev cofactors in HIV-1 RNA translation. Ectopic expression, siRNA knockdown, and trans-complementation assays confirmed that all these cofactors were very important for HIV-1 gene expression and production through Rev and, accordingly, Rev-dependent reporter gene expression. Importantly, these studies revealed for the first time that each of these cofactors also regulated Rev-independent reporter gene expression. To directly determine the roles of these cofactors in HIV-1 RNA translation, we designed and synthesized a full-length capped HIV-1 RNA in vitro, transfected it into cells to bypass the RNA nuclear export step, and determined HIV-1 Gag expression from the cytoplasmic RNA in the cells that had ectopically expressed or siRNA knocked down cofactors. Gag expression was found to closely correlate with the expression levels of all these cofactors. Furthermore, we took advantage of a HIV-1 internal ribosomal entry site (IRES)-based bicistronic reporter gene assay and determined the effects of these cofactors on cap-independent IRES-mediated HIV-1 translation. The results showed that DDX3, eIF5A, and hRIP enhanced HIV-1 IRES-mediated translation, whereas Sam68 did not. Taken together, these results show that HIV-1 Rev cofactors Sam68, eIF5A, hRIP, and DDX3 also function in the translation of HIV-1 RNA and suggest that the regulatory mechanisms of HIV-1 RNA translation are likely different among these cofactors.


Resolving Isomeric Glycopeptide Glycoforms with Hydrophilic Interaction Chromatography (HILIC).
The ability to resolve glycans while attached to tryptic peptides would greatly facilitate glycoproteomics, as this would enable site-specific glycan characterization. Peptide/glycopeptide separations are typically performed using reversed-phase liquid chromatography (RPLC), where retention is driven by hydrophobic interaction. As the hydrophilic glycans do not interact significantly with the RPLC stationary phase, it is difficult to resolve glycopeptides that differ only in their glycan structure, even when these differences are large. Alternatively, glycans interact extensively with the stationary phases used in hydrophilic interaction chromatography (HILIC), and consequently, differences in glycan structure have profound chromatographic shifts in this chromatographic mode. Here, we evaluate HILIC for the separation of isomeric glycopeptide mixtures that have the same peptide backbone but isomeric glycans. Hydrophilic functional groups on both the peptide and the glycan interact with the HILIC stationary phase, and thus, changes to either of these moieties can alter the chromatographic behavior of a glycopeptide. The interactive processes permit glycopeptides to be resolved from each other based on differences in their amino acid sequences and/or their attached glycans. The separations of glycans in HILIC are sufficient to permit resolution of isomeric N-glycan structures, such as sialylated N-glycan isomers differing in α2-3 and α2-6 linkages, while these glycans remain attached to peptides.


Autonomic nervous system activity in acute schizophrenia: I. method and comparison with normal controls.
Many reports of autonomic dysfunction in schizophrenia may have been influenced by the chronicity or medication status of patients, or both. This study eliminates these sources of variation. Skin conductance (SC) and heart rate (HR) base levels and activity were compared in 118 controls and 46 recently admitted, drug-free, acute schizophrenic patients during rest, 20 mild-tone stimuli, and reaction time (RT) and mental arithmetic tasks. Patients showed higher than normal HR and spontaneous SC responses but lower SC base levels. Schizophrenics showed less SC and HR reactivity to tones and RT stimuli, slow habituation of the SC orienting response, an attenuated tonic response to stress, and disproportionately more spontaneous than elicited SC activity. The findings are similar to previous results for unmedicated, chronic schizophrenics and suggest that autonomic activity in schizophrenics is determined relatively more by endogenous factors than by external stimuli.


Error metrics for partially coherent wave fields.
Lensless imaging methods that account for partial coherence have become very common in the past decade. However, there are no metrics in use for comparing partially coherent light fields, despite the widespread use of such metrics to compare fully coherent objects and wave fields. Here, we show how reformulating the mean squared error and Fourier ring correlation in terms of quantum state fidelity naturally generalizes them to partially coherent wave fields. These results fill an important gap in the lensless imaging literature and will enable quantitative assessments of the reliability and resolution of reconstructed partially coherent wave fields.


Examination of health behaviours in a dynamic population.
An increasing USA population, especially as a result of foreign immigration, has been the recent focus of healthcare research and policy debate. However, little is known about domestic immigration and its impact on the measures of health conditions and behaviours used to develop and evaluate public health policies and programmes. The objective of this research is to compare health conditions and behaviours between domestic immigrants and established residents in Nevada, the fastest-growing state in the USA. The Nevada 2004-2006 Behavioural Risk Factor Surveillance Survey and added state residency questions were used to examine the associations between length of residence and 24 explanatory variables on health behaviours and conditions. Weighted multiple logistic regressions were used for data analyses. Among the 8663 respondents weighted to the total Nevada population, long-term residents (5 to <10 years), compared with newcomers or short-term residents, were more likely to report fair or poor health (OR=1.98, 95% CI 1.15 to 3.40) and obesity (OR=2.18, 95% CI 1.10 to 4.32), and to live with firearms in the home (OR=6.34, 95% CI 1.75 to 22.9). They were less likely to report having diabetes (OR=0.49, 95% CI 0.25 to 0.96), having had an HIV test (OR=0.60 95% CI 0.41 to 0.89), having prostate cancer (OR=0.20 95% CI 0.04 to 0.93) or having had a mammogram (OR=0.41, 95% CI 0.25 to 0.70). Long-term residents, compared with newcomers, provide a unique comparison group that share the same physical and policy environment but in different time periods. Knowledge of differences such as these can improve the specificity in identifying target groups for particular interventions.


Silica@zirconia@poly(malic acid) nanoparticles: promising nanocarriers for theranostic applications.
Silica@zirconia@poly(malic acid) nanocarriers of 110 nm mean diameter were designed, synthesized and characterized for the targeted delivery of diagnostic and therapeutic 99mTc to folate-overexpressing tumors. An important achievement was that a multifunctional l-(-)-malic-acid-based copolymer was formed in situ at the surface of the inorganic cores in a single synthetic step incorporating l-(-)-malic acid, β-cyclodextrin rings, folic acid moieties, and polyethylene glycol chains. Morphological and in-depth structural analysis of the particles proved their core@shell structure. Stability experiments in aqueous media evidenced that stable suspensions can be obtained from the lyophilized powder in 10 mM phosphate buffer at pH 7.4. During 14-day degradation experiments, the nanoparticles were found to be slowly dissolving (including inorganic core) in saline and also in total cell medium. An in vitro toxicity assay on hepatocytes showed a concentration-dependent decrease of cell viability down to 63 ± 1% at the highest applied concentration (0.5 mg ml-1). Proof of concept experiments of technetium-99m radiolabelling and in vivo labelling stability are presented.


RNAi-dependent and independent control of LINE1 accumulation and mobility in mouse embryonic stem cells.
In most mouse tissues, long-interspersed elements-1 (L1s) are silenced via methylation of their 5'-untranslated regions (5'-UTR). A gradual loss-of-methylation in pre-implantation embryos coincides with L1 retrotransposition in blastocysts, generating potentially harmful mutations. Here, we show that Dicer- and Ago2-dependent RNAi restricts L1 accumulation and retrotransposition in undifferentiated mouse embryonic stem cells (mESCs), derived from blastocysts. RNAi correlates with production of Dicer-dependent 22-nt small RNAs mapping to overlapping sense/antisense transcripts produced from the L1 5'-UTR. However, RNA-surveillance pathways simultaneously degrade these transcripts and, consequently, confound the anti-L1 RNAi response. In Dicer(-/-) mESC complementation experiments involving ectopic Dicer expression, L1 silencing was rescued in cells in which microRNAs remained strongly depleted. Furthermore, these cells proliferated and differentiated normally, unlike their non-complemented counterparts. These results shed new light on L1 biology, uncover defensive, in addition to regulatory roles for RNAi, and raise questions on the differentiation defects of Dicer(-/-) mESCs.


Synergistic Effects of Pyrrolic N/Pyridinic N on Ultrafast Microwave Synthesized Porous CoP/Ni2P to Boost Electrocatalytic Hydrogen Generation.
Transition metal phosphides are ideal inexpensive electrocatalysts for water-splitting, but the catalytic activity still falls behind that of noble metal catalysts. Therefore, developing valid strategies to boost the electrocatalytic activity is urgent to promote large-scale applications. Herein, a microwave combustion strategy (20 s) is applied to synthesize N-doped CoP/Ni2P heterojunctions (N-CoP/Ni2P) with porous structure. The porous structure expands the specific surface area and accelerates the mass transport efficiency. Importantly, the pyrrolic N/pyridinic N content is adjusted by changing the amount of urea during the synthesis process and then optimizing the adsorption/desorption capacity for H*/OH* to enhance the catalyst activity. Then, the synthesized N-CoP/Ni2P exhibits small overpotentials of 111 and 133 mV for HER in acidic and alkaline electrolytes and 290 mV for OER in alkaline electrolytes. This work provides an original and efficient approach to the synthesis of porous metal phosphides.


Average effect of a mutation in lignin biosynthesis in loblolly pine.
Cinnamyl alcohol dehydrogenase (CAD, E.C. 184.108.40.206) is a monolignol biosynthetic enzyme that catalyzes the final step of lignin subunit biosynthesis in higher plants. Recently, a mutant allele of the cad gene, cad-n1, encoding for the CAD enzyme, was discovered in loblolly pine. By reducing the expression of the cad gene, this mutant has a decreased lignin content and major changes in the lignin composition in wood. In this study, we found that the substitution of a wild-type allele by cad-n1 was associated with a significant effect on 2nd-year shoot elongation in a half-sib family of loblolly pine (designated family 7-1037). The average effect of cad-n1 appeared to increase with tree growth and was greater for stem radial growth than height growth. An increase of 14.1% in de-barked volume in year 4 was associated with cad-n1. Co-segregation analysis indicated that the cad locus itself might represent a gene that governs stem growth in pine. The significance of the mutation cad-n1 for tree growth and wood processing is discussed.


Effect of radial keratotomy incision direction on wound depth.
The majority of ophthalmic surgeons who perform radial keratotomy make incisions from the optical clear zone to the limbus (downhill; centrifugal), instead of from the limbus to the optical clear zone (uphill; centripetal). To compare the efficacy of these techniques, one surgeon performed keratotomy incisions in 10 eyes of 5 monkeys using the same double-edged diamond blade set to 80% of central pachometry. Four or eight centrifugal (downhill) and centripetal (uphill) incisions were made in each eye. Achieved incision depth was measured by light microscopy 2.5 to 8 months postoperative. Downhill incision depth averaged 46% (range 38% to 61%), whereas uphill incision depth averaged 74% (range 53% to 87%) (P less than .0005). In both groups, incision deviation from the perpendicular (lateral tilt error) was greatest adjacent to the optical clear zone (P less than .01). These differences may be explained by tilt error (forward or backward) or by the perpendicular front cutting action of a vertical blade being more effective than the angled blade.


Naringin targets Zeb1 to suppress osteosarcoma cell proliferation and metastasis.
Naringin, a citrus bioflavonoid, has anti-inflammatory actions and cardio- and neuroprotective effects. In addition, naringin exhibits multiple antitumor actions in several cancer types, including osteosarcoma, the most common type of bone cancer. Here, we show that naringin inhibits proliferation and invasion and induces apoptosis in human osteosarcoma cells by inhibiting zinc finger E-box binding homeobox 1 (Zeb1), a transcriptional repressor of epithelial differentiation involved in tumor metastasis. Our expression analyses confirm that Zeb1 is highly expressed in osteosarcoma specimens and cell lines. The effects of naringin, which included downregulation of Cyclin D1, MMP2, and bcl-2, where reproduced by siRNA-mediated Zeb1 silencing, whereas Zeb1 overexpression increased proliferation, migration, and Cyclin D1, MMP2, and bcl-2 levels. In addition, naringin administration reduced tumor nodule formation and attenuated the expression of the above proteins in the livers of mice injected with MG63 osteosarcoma cells. Our study provides preclinical evidence for the potential therapeutic application of naringin in the treatment of osteosarcoma.


Characterization of a linear streak artifact with pulse inversion tissue harmonics in musculoskeletal sonography.
To understand a linear artifact that projects deep to reflective structures that move rapidly while using tissue harmonic imaging with pulse inversion (PI) sonography. We hypothesize that this artifact is due to a cancellation error between firings in PI imaging, and it is, therefore, similar in generation to the twinkling artifact in color Doppler sonography. This artifact could be studied with the use of surfaces of different roughness to represent different rates of motion, in which roughness corresponds to spatial fluctuations in surface height. Given very slight variations in beam focusing as occurs with sonographic imaging arrays, these spatial fluctuations translate into temporal fluctuations in the received signal as would occur with tissue motion. We scanned 4 different sandpaper grits and a smooth surface through a water path using fundamental and PI mode, 1- and 2-pulse techniques, respectively. The sandpaper and the smooth surface were scanned through a water path at mechanical indices of 0.1 to 0.7. Four independent images were subtracted pairwise to remove nonfluctuating signals. These noise pixels were counted and analyzed. Analysis of variance showed that the noise generated behind the different surfaces was highly significantly different. Two-tailed t tests generally showed significant differences in the quantity of noise between fundamental and harmonic imaging behind the roughest 3 grades of sandpaper. A multiple regression model showed significantly greater slopes for harmonic imaging for all grades of sandpaper and the smooth surface. The noise and, by extension, the linear streak artifact in musculoskeletal imaging are dependent on the mechanical index and are functions of sandpaper roughness. This would be equivalent to a subtraction error between 2 firings due to soft tissue motion, and the artifact may be a way to identify rapid soft tissue motion in PI images.


Crystal Structures of Quinolinate Synthase in Complex with a Substrate Analogue, the Condensation Intermediate, and Substrate-Derived Product.
The enzyme NadA catalyzes the synthesis of quinolinic acid (QA), the precursor of the universal nicotinamide adenine dinucleotide (NAD) cofactor. Here, we report the crystal structures of complexes between the Thermotoga maritima (Tm) NadA K219R/Y107F variant and (i) the first intermediate (W) resulting from the condensation of dihydroxyacetone phosphate (DHAP) with iminoaspartate and (ii) the DHAP analogue and triose-phosphate isomerase inhibitor phosphoglycolohydroxamate (PGH). In addition, using the TmNadA K219R/Y21F variant, we have reacted substrates and obtained a crystalline complex between this protein and the QA product. We also show that citrate can bind to both TmNadA K219R and its Y21F variant. The W structure indicates that condensation causes dephosphorylation. We propose that catalysis by the K219R/Y107F variant is arrested at the W intermediate because the mutated protein is unable to catalyze its aldo-keto isomerization and/or cyclization that ultimately lead to QA formation. Intriguingly, PGH binds to NadA with its phosphate group at the site where the carboxylate groups of W also bind. Our results shed significant light on the mechanism of the reaction catalyzed by NadA.


Metabolomics screening of serum biomarkers for occupational exposure of titanium dioxide nanoparticles.
Although nanotoxicology studies have shown that respiratory exposure of titanium dioxide nanoparticles (TiO2 NPs) could induce adverse health effects, limited biomarkers associated with occupational exposure of TiO2 NPs were reported. The purpose of this study is to screen serum biomarkers among workers occupationally exposed to TiO2 NPs using metabolomics. Compared with the control group, a total of 296 serum metabolites were differentially expressed in the TiO2 NPs-exposed group, of which the relative expression of 265 metabolites increased, and the remaining 31 decreased. Three machine learning methods including random forest (RF), support vector machines (SVM), and boruta screened eight potential biomarkers and simultaneously selected a metabolite, Liquoric acid. Through multiple linear regression analysis to adjust the influence of confounding factors such as gender, age, BMI, smoking and drinking, occupational exposure to TiO2 NPs was significantly related to the relative expression of the eight potential biomarkers. Meanwhile, the receiver operating characteristic curves (ROCs) of these potential biomarkers had good sensitivity and specificity. These potential biomarkers were related to lipid peroxidation, and had biological basis for occupational exposure to TiO2 NPs. Therefore, it was demonstrated that the serum metabolites represented by Liquoric acid were good biomarkers of occupational exposure to TiO2 NPs.


An air suspension to demonstrate the properties of torsion balances with fibers of zero length.
We report on the design and characterization of an air-bearing suspension that has been constructed to highlight the properties of torsion balances with fibers of zero length. A float is levitated on this suspension, and its rotational and translational motion in the horizontal plane of the laboratory is controlled using magnetic actuators. We demonstrate the in situ electromagnetic tuning of the float's center-of-buoyancy to an accuracy of ±0.3 mm, which was limited by the noise in the air bearing. The rotational stiffness of the float, which is approximately zero by design, was also measured. We compare the observed behavior of the float with the predictions of a detailed model of the statics of the float-actuator system. Finally, we briefly discuss the application of these ideas and results to the construction of sensitive devices for the measurement of weak forces with short ranges.


Beneficial effects of exogenous tetrahydrobiopterin on left ventricular remodeling after myocardial infarction in rats: the possible role of oxidative stress caused by uncoupled endothelial nitric oxide synthase.
Reactive oxygen species (ROS) is deeply involved in the process of ventricular remodeling after myocardial infarction (MI). Under oxidative stress, endothelial nitric oxide synthase (eNOS) can be converted to a ROS generator, because a relative lack of tetrahydrobiopterin (BH4), an essential cofactor for NO synthesis, leads to eNOS uncoupling. The uncoupled eNOS generates superoxide rather than NO. The possible role of ROS generated by eNOS in ventricular remodeling after MI was investigated. Rats were treated with oral BH4 supplementation starting at 3 days before coronary artery ligation. At 4 weeks after MI, there was augmented superoxide production in association with reduced BH4/dihydrobiopterin (BH2) ratio and eNOS dimer/monomer protein ratio in the heart. Treatment with BH4 increased BH4/BH2 ratio and eNOS dimer/monomer ratio, and decreased superoxide production. In BH4-treated MI rats, left ventricular size was smaller, thickness of the non-infarcted posterior wall was thinner, and cardiac function was preserved compared with the control MI rats. The present study suggested that ventricular remodeling process after MI leads to BH4 oxidation and resulted in uncoupled eNOS-derived superoxide generation, which further augmented the remodeling process and deteriorated cardiac function.