Identification of key genes and pathways in hepatocellular carcinoma: A preliminary bioinformatics analysis.
Hepatocellular carcinoma (HCC) is one of the most prevalent cancers worldwide. However, the precise mechanisms of the development and progression of HCC remain unclear. The present study attempted to identify and functionally analyze the differentially expressed genes between HCC and cirrhotic tissues by using comprehensive bioinformatics analyses. The GSE63898 gene expression profile was downloaded from the Gene Expression Omnibus (GEO) and analyzed using the online tool GEO2R to identify differentially expressed genes (DEGs). Gene ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the DEGs were performed in DAVID. The STRING database was used to evaluate the interactions of DEGs and to construct a protein-protein interaction (PPI) network using Cytoscape software. Hub genes were selected using the cytoHubba plugin and were validated with the cBioPortal database. A total of 301 DEGs were identified between HCC and cirrhotic tissues. The GO analysis results showed that these DEGs were significantly enriched in certain biological processes including negative regulation of growth and cell chemotaxis. Several significant pathways, including the p53 signaling pathway, were identified as being closely associated with these DEGs. The top 12 hub genes were screened and included TTK, NCAPG, TOP2A, CCNB1, CDK1, PRC1, RRM2, UBE2C, ZWINT, CDKN3, AURKA, and RACGAP1. The cBioPortal analysis found that alterations in hub genes could result in significantly reduced disease-free survival in HCC. The present study identified a series of key genes and pathways that may be involved in the tumorigenicity and progression of HCC, providing a new understanding of the underlying molecular mechanisms of carcinogenesis in HCC.


Dispersibility of carbon dots in aqueous and/or organic solvents.
Carbon dots have a wide range of applications in biological and medical fields as an alternative to quantum dots because of their low toxicity and excellent luminescence properties. To date, a large number of carbon dots have been prepared and they were consequently reviewed according to their synthetic method, luminescence properties and related applications. The dispersibility of carbon dots in aqueous and/or organic solvents could actually play a significant role in the properties and various application fields, however, such a perspective is ignored by most of the reviewed literature studies. Thus in this minireview, we focus on the surfactant groups of carbon dots which could be classified as hydrophilic, hydrophobic and amphiphilic types. They have accordingly a marked impact on the dispersibility of carbon dots in different solvents as well as the further advantage in those imaging studies in vitro and in vivo.


Developmental stage oxidoreductive states of Chlamydia and infected host cells.
A defining characteristic of Chlamydia spp. is their developmental cycle characterized by outer membrane transformations of cysteine bonds among cysteine-rich outer membrane proteins. The reduction-oxidation states of host cell compartments were monitored during the developmental cycle using live fluorescence microscopy. Organelle redox states were studied using redox-sensitive green fluorescent protein (roGFP1) expressed in CF15 epithelial cells and targeted to the cytosol, mitochondria, and endoplasmic reticulum (ER). The redox properties of chlamydiae and the inclusion were monitored using roGFP expressed by Chlamydia trachomatis following transformation. Despite the large morphological changes associated with chlamydial infection, redox potentials of the cytosol (Ψ(cyto) [average, -320 mV]), mitochondria (Ψ(mito) [average, -345 mV]), and the ER (ΨER [average, -258 mV]) and their characteristic redox regulatory abilities remained unchanged until the cells died, at which point Ψ(cyto) and Ψ(mito) became more oxidized and Ψ(ER) became more reduced. The redox status of the chlamydial cytoplasm was measured following transformation and expression of the roGFP biosensor in C. trachomatis throughout the developmental cycle. The periplasmic and outer membrane redox states were assessed by the level of cysteine cross-linking of cysteine-rich envelope proteins. In both cases, the chlamydiae were highly reduced early in the developmental cycle and became oxidized late in the developmental cycle. The production of a late-developmental-stage oxidoreductase/isomerase, DsbJ, may play a key role in the regulation of the oxidoreductive developmental-stage-specific process. Infectious Chlamydia organisms have highly oxidized and cysteine cross-linked membrane proteins that confer environmental stability when outside their host cells. Once these organisms infect a new host cell, the proteins become reduced and remain reduced during the active growth stage. These proteins become oxidized at the end of their growth cycle, wherein infectious organisms are produced and released to the environment. How chlamydiae mediate and regulate this key step in their pathogenesis is unknown. Using biosensors specifically targeted to different compartments within the infected host cell and for the chlamydial organisms themselves, the oxidoreductive states of these compartments were measured during the course of infection. We found that the host cell redox states are not changed by infection with C. trachomatis, whereas the state of the chlamydial organisms remains reduced during infection until the late developmental stages, wherein the organisms' cytosol and periplasm become oxidized and they acquire environmental resistance and infectivity.


Revealing protein-protein interactions at the transcriptome scale by sequencing.
We describe PROPER-seq (protein-protein interaction sequencing) to map protein-protein interactions (PPIs) en masse. PROPER-seq first converts transcriptomes of input cells into RNA-barcoded protein libraries, in which all interacting protein pairs are captured through nucleotide barcode ligation, recorded as chimeric DNA sequences, and decoded at once by sequencing and mapping. We applied PROPER-seq to human embryonic kidney cells, T lymphocytes, and endothelial cells and identified 210,518 human PPIs (collected in the PROPER v.1.0 database). Among these, 1,365 and 2,480 PPIs are supported by published co-immunoprecipitation (coIP) and affinity purification-mass spectrometry (AP-MS) data, 17,638 PPIs are predicted by the prePPI algorithm without previous experimental validation, and 100 PPIs overlap human synthetic lethal gene pairs. In addition, four previously uncharacterized interaction partners with poly(ADP-ribose) polymerase 1 (PARP1) (a critical protein in DNA repair) known as XPO1, MATR3, IPO5, and LEO1 are validated in vivo. PROPER-seq presents a time-effective technology to map PPIs at the transcriptome scale, and PROPER v.1.0 provides a rich resource for studying PPIs.


Bismuth-rich bismuth oxyiodide microspheres with abundant oxygen vacancies as an efficient photocatalyst for nitrogen fixation.
Solar-driven reduction of dinitrogen (N2) to ammonia (NH3) is still challenging due to the highly stable N-N triple bond. Herein, orthorhombic phase H-Bi5O7I microspheres with abundant oxygen vacancies (OVs) were successfully prepared via a simple calcination and hydrogen reduction strategy. Based on the combined bismuth-rich strategy and the introduction of OVs in H-Bi5O7I, a remarkable photocatalytic nitrogen fixation performance was achieved under visible light irradiation in the absence of any organic scavengers or noble-metal cocatalysts. H-Bi5O7I exhibits an NH4+ generation rate of 162.48 μmol g-1 h-1, which is 2.0 and 7.4 times higher than that of Bi5O7I and BiOI. It is because H-Bi5O7I with abundant OVs has a more proper band gap, better electron capturing ability and more effective separation and transfer rate for the photogenerated charge carriers. This study may provide a new direction and useful insight for the design of defect assisted N2 fixation photocatalysts.


In-situ, time resolved monitoring of uranium in BFS:OPC grout. Part 2: Corrosion in water.
To reflect potential conditions in a geological disposal facility, uranium was encapsulated in grout and submersed in de-ionised water for time periods between 2-47 weeks. Synchrotron X-ray Powder Diffraction and X-ray Tomography were used to identify the dominant corrosion products and measure their dimensions. Uranium dioxide was observed as the dominant corrosion product and time dependent thickness measurements were used to calculate oxidation rates. The effectiveness of physical and chemical grout properties to uranium corrosion and mobilisation is discussed and Inductively Coupled Plasma Mass Spectrometry was used to measure 238U(aq) content in the residual water of several samples.


[The circahoral rhythm of the functional activity of rat adrenocorticocytes].
Functional manifestations of biological ultradian rhythms in the adrenal cortex were first confirmed on the cytologic level. The cycle of the ultradian rhythm in the cells of the adrenal cortex fascicular zone consisted was determined to involve three phases: accumulation of lipids, delipoidization, and the rest period. A definite volume of cells and their nuclei, certain nuclear-cytoplasmic relations, the number of liposomes and mitochondria on the square unit are characteristic of each phase. The activity of cells is especially high during the second phase which is characterized by the increase in nuclear sizes and nuclear-cytoplasmatic relations, by the decrease in cell volume and in the number of liposomes in their cytoplasm; this is indicative of the rised synthesis and secretion of hormones. It is underlined that the cell volume increases when liposomes are accumulated in the cytoplasm to decrease upon their disappearance.


Recording of Influenza-Like Illness in UK Primary Care 1995-2013: Cohort Study.
There is a lack of recent studies examining recording of influenza-like illness (ILI) in primary care in the UK over time and according to population characteristics. Our aim was to determine time trends and socio-demographic patterns of ILI recorded consultations in primary care. We used The Health Improvement Network (THIN) UK primary care database and extracted data on all ILI consultations between 1995 and 2013. We estimated ILI recorded consultation rates per 100,000 person-weeks (pw) by age, gender, deprivation and winter season. Negative binomial regression models were used to examine time trends and the effect of socio-demographic characteristics. Trends in ILI recorded consultations were compared to trends in consultations with less specific symptoms (cough or fever) recorded. The study involved 7,682,908 individuals in 542 general practices. The ILI consultation rate decreased from 32.5/100,000 pw (95% confidence interval (CI) 32.1, 32.9) in 1995-98 to 15.5/100,000 pw (95% CI 15.4, 15.7) by 2010-13. The decrease occurred prior to 2002/3, and rates have remained largely stable since then. Declines were evident in all age groups. In comparison, cough or fever consultation rates increased from 169.4/100,000 pw (95% CI 168.6, 170.3) in 1995-98 to 237.7/100,000 pw (95% CI 237.2, 238.2) in 2010-13. ILI consultation rates were highest among individuals aged 15-44 years, higher in women than men, and in individuals from deprived areas. There is substantial variation in ILI recorded consultations over time and by population socio-demographic characteristics, most likely reflecting changing recording behaviour by GPs. These results highlight the difficulties in using coded information from electronic primary care records to measure the severity of influenza epidemics across time and assess the relative burden of ILI in different population subgroups.


Update on laboratory studies and relationship to rheumatic and allergic diseases.
Guidelines have been presented for the optimal selection and interpretation of various laboratory tests related to rheumatologic, immunologic, and allergic disorders. The last decade has witnessed many technologic changes in the performance of screening tests for autoantibodies, and an expeditious approach to the detection of the clinically useful autoantibody system has been outlined. When both an immunofluorescent assay using an actively dividing substrate such as HEp-2 and an immunodiffusion test for precipitin autoantibodies are performed on patient sera, it is likely that most of the significant diagnostic and prognostic systems will be detected. Although exact pathogenetic roles have not been defined for all autoantibodies, it is apparent that highly specific markers for certain rheumatic diseases exist and the pertinent clinical aspects of each individual autoantibody-autoantigen system has been summarized. Why so many autoantibodies are cross-reactive with seemingly unrelated antigens is a perplexing aspect of autoimmunity, a disease process for which the necessary and sufficient conditions of development have not been defined even after decades of research. One recent hypothesis regarding the generation of diverse autoantibodies is the tenet that production is interrelated in an auto-anti-idiotypic network. Support for this hypothesis comes from the recent successful use of anti-idiotypes to DNA autoantibodies in an attempt to abrogate pathogenetic effects of target organ damage in murine models of SLE. A similar approach might be undertaken with some of the other autoantibody systems in an attempt at a novel therapy for disorders of immunoregulation.


Influence of acetylene on growth of sulfate-respiring bacteria.
At a concentration of 20% of the atmosphere of the culture flasks, acetylene inhibited growth and carbon dioxide production by Desulfovibrio desulfuricans and Desulfovibrio gigas. The bacteria did not reduce acetylene to ethylene, and neither acetylene dicarboxylic acid nor ethylene was inhibitory. At 10%, acetylene was partially inhibitory for the desulfovibrios. At 5%, acetylene impeded the rate but did not limit the extent of growth and catabolism of the desulfovibrios. Desulfotomaculum ruminis was affected only negligibly, if at all, by acetylene and ethylene at any of these concentrations.


Hypertension and Atrial Fibrillation: Doubts and Certainties From Basic and Clinical Studies.
Hypertension and atrial fibrillation (AF) are 2 important public health priorities. Their prevalence is increasing worldwide, and the 2 conditions often coexist in the same patient. Hypertension and AF are strikingly related to an excess risk of cardiovascular disease and death. Hypertension ultimately increases the risk of AF, and because of its high prevalence in the population, it accounts for more cases of AF than other risk factors. Among patients with established AF, hypertension is present in about 60% to 80% of individuals. Despite the well-known association between hypertension and AF, several pathogenetic mechanisms underlying the higher risk of AF in hypertensive patients are still incompletely known. From an epidemiological standpoint, it is unclear whether the increasing risk of AF with blood pressure (BP) is linear or threshold. It is uncertain whether an intensive control of BP or the use of specific antihypertensive drugs, such as those inhibiting the renin-angiotensin-aldosterone system, reduces the risk of subsequent AF in hypertensive patients in sinus rhythm. Finally, in spite of the observational evidence suggesting a progressive relation between BP levels and the risk of thromboembolism and bleeding in patients with hypertension and AF, the extent to which BP should be lowered in these patients, including those who undergo catheter ablation, remains uncertain. This article summarizes the main basic mechanisms through which hypertension is believed to promote AF. It also explores epidemiological data supporting an evolutionary pathway from hypertension to AF, including the emerging evidence favoring an intensive BP control or the use of drugs, which inhibit the renin-angiotensin-aldosterone system to reduce the risk of AF. Finally, it examines the impact of non-vitamin K antagonist oral anticoagulants compared with warfarin in relation to hypertension.


Heparin-stimulated inhibition of factor IXa generation and factor IXa neutralization in plasma.
Generation and inhibition of activated factor IXa was studied in factor XIa-activated plasma containing 4 mmol/L free calcium ions and 20 mumol/L phospholipid (25 mol% phosphatidylserine/75 mol% phosphatidylcholine). Interference of other (activated) clotting factors with the factor IXa activity measurements could be avoided by using a highly specific and sensitive bioassay. Factor IXa generation curves were analyzed according to a model that assumed Michaelis-Menten kinetics of factor XIa-catalyzed factor IXa formation and pseudo first order kinetics of inhibition of factor XIa and factor IXa. In the absence of heparin, factor IXa activity in plasma reached final levels that were found to increase with increasing amounts of factor XIa used to activate the plasma. When the model was fitted to this set of factor IXa generation curves, the analysis yielded a rate constant of inhibition of factor XIa of 0.7 +/- 0.1 min-1 and a kcat/Km ratio of 0.29 +/- 0.01 (nmol/L)-1 min-1. No neutralization of factor IXa activity was observed (the estimated rate constant of inhibition of factor IXa was 0). Thus, in the absence of heparin, the final level of factor IXa in plasma is only dependent on the initial factor XIa concentration. While neutralization of in situ generated factor IXa in normal plasma was negligible, unfractionated heparin dramatically enhanced the rate of inactivation of factor IXa (apparent second order rate constant of inhibition of 5.2 min-1/per microgram heparin/mL). The synthetic pentasaccharide heparin, the smallest heparin chain capable of binding antithrombin III, stimulated the inhibition of in situ generated factor IXa, but sevenfold less than unfractionated heparin (k = 0.76 min-1 per microgram pentasaccharide/mL). We found that free calcium ions were absolutely required to observe an unfractionated heparin and pentasaccharide-stimulated neutralization of factor IXa activity. Factor XIa inhibition (psuedo first order rate constant of 0.7 min-1) was not affected by unfractionated heparin or pentasaccharide in the range of heparin concentrations studied.


[Effect of polyphenols from the Mediterranean diet on proliferation and mediators of in vitro invasiveness of the MB-49 murine bladder cancer cell line].
To evaluate the effect of different polyphenols on the proliferation and invasive capacity of MB-49 murine bladder tumor cell lines and to identify the mediators involved in this process. MB-49 murine bladder cancer cells were cultured in media supplemented with resveratrol, rutin, morin, quercetin, gallic acid and tannic acid (all of them are polyphenols usually present in Mediterranean diet) for periods of 24, 48 and 72 hours to quantify the expression of urokinase-type plasminogen activator (uPA) and its receptor (uPAR) in the culture medium, as well as of metalloproteinase-9 (MMP-9) and cell proliferation. All the polyphenols studied significantly inhibited proliferation of MB-49 cells, varying according to the time periods and doses used. The cells in the media supplemented with the nutrients to study did not show inhibition of mRNA expression of urokinase-type plasminogen activator (uPA) or its high affinity receptor (uPAR). It was even slightly increased in certain cases. However, mRNA expression of metalloproteinase-9 was strongly inhibited. The polyphenols present in our usual diet exert an effect on the proliferation and mediators of bladder tumor invasiveness in MB-49 cells.


Socioeconomic Status, Smoking, Alcohol use, Physical Activity, and Dietary Behavior as Determinants of Obesity and Body Mass Index in the United States: Findings from the National Health Interview Survey.
The aim of this research was to study the socio-demographic and behavioral determinants of obesity and Body Mass Index (BMI) in the United States, using a nationally representative sample. We used data from the 2010 US National Health Interview Survey. Analyses were limited to adults 18 years and older (N=23,434). Multivariate regression analyses were conducted to estimate the associations between covariates and obesity and BMI. Overall, 28.1% in the sample were obese and the mean BMI was 27.6 kg/m(2). In adjusted models, we found that older age, non-Hispanic Black race, lower education and income levels, Midwestern and Southern region of residence, former smoking, infrequent alcohol use, physical inactivity, consumption of less fruits, vegetables, brown rice and more cheese, fried potato and meat, were associated with obesity. These factors were also associated with higher BMI, along with male gender and higher consumption of meat, fried potatoes and cheese. The association of many of the socio-demographic and behavioral factors with obesity and higher BMI found in our study was consistent with previous findings. Persistence of such associations suggest a need for better understanding of the underlying mechanism as well as for evaluation of the current programs and policies targeted at reducing the obesity burden in the United States. In view of the rising global obesity epidemic, especially in the low- and middle-income countries, our findings could help guide development of effective health and social policies and programs aimed at reducing the obesity burden in other parts of the world.


Molecular cloning and characterization of ficolin, a multimeric protein with fibrinogen- and collagen-like domains.
We have previously identified and purified transforming growth factor-beta 1 (TGF-beta 1)-binding proteins from porcine uterus membranes (Ichijo, H., Rönnstrand, L., Miyagawa, K., Ohashi, H., Heldin, C.-H., and Miyazono, K. (1991) J. Biol. Chem. 266, 22459-22464). One of these TGF-beta 1-binding proteins, with a molecular weight of 40,000, was purified to homogeneity and subjected to amino acid sequence analysis. The amino acid sequences obtained were used to isolate two closely related cDNA clones from a porcine uterus cDNA library. The deduced amino acid sequences revealed that both cDNAs encoded proteins that were mainly composed of fibrinogen-like and collagen-like domains. Therefore, they were denoted ficolin-alpha and ficolin-beta. Expression of ficolin-alpha and -beta cDNA in mammalian cells revealed that ficolin forms dimers, trimers, and several higher order of oligomers, whose molecular weights fit well with those of the purified TGF-beta 1-binding proteins from porcine uterus. Moreover, immunoblotting analysis using a peptide anti-serum against ficolin indicated that the TGF-beta 1-binding proteins identified in porcine uterus are ficolin-alpha, -beta, and their oligomers or closely related molecules. However, recombinant ficolin-alpha and -beta did not bind TGF-beta 1, despite the similarities in molecular weights and immunoreactivity with the material from the natural source. It is possible that a specific posttranslational modification of ficolin or interaction with another component is needed for TGF-beta 1 binding. Analysis by Northern blotting revealed that the expression of ficolin-alpha mRNA is relatively restricted and most abundant in placenta and lung. On the other hand, ficolin-beta was mainly expressed in skeletal muscle. The in vivo functions of ficolin will be discussed.


Bortezomib downregulates MGMT expression in T98G glioblastoma cells.
The efficacy of treatment for glioblastoma multiforme is currently limited by the development of resistance, particularly, but not exclusively, due to the expression of the DNA repair enzyme O6-methylguanine methyltransferase (MGMT) in a significant proportion of astrocytic tumors. MGMT is post-translationally regulated by the 26S proteasome, a multi-subunit organelle responsible for degradation of misfolded cellular proteins. The boronic acid dipeptide bortezomib is the first and only proteasome inhibitor in clinical use so far, and has been reported as a strategy to restrict growth and promote apoptosis of glioblastoma cells. In this study we investigated the effect of bortezomib on MGMT expression in T98G cells, looking for an effect on the nuclear factor kappa B (NFκB) pathway, which is a major player in MGMT regulation and is also under tight control by the ubiquitin-proteasome system. Administration of bortezomib led to a significant reduction of T98G cell viability and induction of DNA fragmentation. These effects coincided with reduced expression of MGMT transcript levels, and a decrease in cellular amount and IκBα-mediated, proteasomal activity-dependent nuclear translocation of NFκB. In addition, bortezomib-induced phosphorylation of the translation initiation factor 2alpha (eIF2α) was in parallel with translational repression of MGMT. Taken together, these results suggest a novel role for bortezomib as a potent MGMT inhibitor and support its ongoing testing as a chemosensitizer in glioblastoma.


Alkaline modification of the acid residue of incinerated sewage sludge ash after phosphorus recovery for heavy metal removal from aqueous solutions.
Enriched in phosphorus, sewage sludge ash has been extensively studied and applied as a secondary source for phosphorus recovery. Wet extraction, especially acid washing, is one of the most feasible methods to recover phosphorus from the ash due to its ease of operation, high efficiency and low cost. However, the management of the resultant acid residue was seldom addressed. In this study, special focus was paid to the reuse and recycling of the acid residue by an alkaline activation method. Its adsorption performance towards four different heavy metals in aqueous solutions was evaluated by batch and fixed-bed column adsorption experiments. The obtained material showed a high BET specific area (98.29 m2/g) and a total pore volume (0.114 cm3/g), and effectively removed Cd(II), Cu(II), Pb(II) and Zn(II) from aqueous solutions with the maximum adsorption capacity of around 26.8, 22.2, 53.3 and 13.5 mg/g respectively. It could be loaded in a fixed-bed column to continuously remove heavy metals especially for Pb(II). The proposed method to recycle the acid residue makes the wet extraction methods designing to recover phosphorus from incinerated sewage sludge complete without the generation of waste, which contributes to circular economy and a sustainable future.


Evaluation of Ischemic and Bleeding Risks Associated With 2 Parenteral Antiplatelet Strategies Comparing Cangrelor With Glycoprotein IIb/IIIa Inhibitors: An Exploratory Analysis From the CHAMPION Trials.
In the context of contemporary pharmacotherapy, optimal antiplatelet management with percutaneous coronary intervention (PCI) has not been well established. To compare the ischemic and bleeding risks associated with glycoprotein IIb/IIIa inhibitors (GPIs) and a potent P2Y12 antagonist, cangrelor, in patients undergoing PCI. An exploratory analysis of pooled patient-level data from the 3 phase 3 Cangrelor vs Standard Therapy to Achieve Optimal Management of Platelet Inhibition (CHAMPION PCI, CHAMPION PLATFORM, and CHAMPION PHOENIX) trials of patients undergoing elective or nonelective PCI. The participants included 10 929 patients assigned to cangrelor but not receiving GPIs (cangrelor alone) and 1211 patients assigned to clopidogrel (or placebo) and receiving routine GPIs (clopidogrel-GPI). Patients requiring bailout or rescue GPI therapy were excluded. To account for risk imbalances, 1:1 propensity score matching based on 16 baseline clinical variables yielded 1021 unique matched pairs. The present study's data analysis was conducted from October 28, 2015, to August 6, 2016. The primary efficacy end point was the composite of all-cause mortality, myocardial infarction, ischemia-driven revascularization, or stent thrombosis at 48 hours. Safety was assessed by 3 validated bleeding scales (Global Use of Strategies to Open Occluded Coronary Arteries [GUSTO], Thrombolysis in Myocardial Infarction [TIMI], and Acute Catheterization and Urgent Intervention Triage) and requirement for blood transfusions. Of the 12 140 patients included in the analysis, 8779 were men (72.3%), and the mean (SD) age was 63.2 (11.3) years. Patients in the clopidogrel-GPI group were more likely to be male (75.6% vs 71.9%), younger (median, 60 [range, 23-91] years vs 64 [range, 26-95] years), enrolled from the United States (77.9% vs 40.0%), and present with an acute coronary syndrome, but they had lower comorbid disease burden and were less likely to receive bivalirudin (8.8% vs 27.3%). In the matched cohorts, the rates of the primary efficacy end point were not significantly different between the cangrelor alone and clopidogrel-GPI groups (2.6% vs 3.3%; odds ratio [OR], 0.79; 95% CI, 0.48-1.32). There was a nonsignificant trend toward lower rates of GUSTO-defined severe/life-threatening bleeding with cangrelor alone compared with clopidogrel-GPI (0.3% vs 0.7%; OR, 0.43; 95% CI, 0.11-1.66). Rates of TIMI-defined major or minor bleeding were significantly lower in patients treated with cangrelor alone (0.7% vs 2.4%; OR, 0.29; 95% CI, 0.13-0.68). Based on a pooled analysis from the 3 phase 3 CHAMPION trials, cangrelor alone was associated with similar ischemic risk and lower risk-adjusted bleeding risk compared with clopidogrel-GPIs. clinicaltrials.gov Identifiers: NCT00305162, NCT00385138, and NCT01156571.


The prevalence of atypical serum cholinesterase in a Nigerian population.
The prevalence of the atypical phenotype of pseudocholinesterase was looked for in 345 individuals living in Zaria district of Northern Nigeria by the differential inhibitory actions of dibucaine and sodium fluoride. The frequency of the atypical gene, Ea1 was found to be 0.9% and the frequency of the fluoride insensitive phenotype Ef1, was 0.74%. The atypical enzyme or the intermediate phenotype was not observed in any of the 42 patients with sickle-cell disease.


A bilayered nanoshell for durable protection of single yeast cells against multiple, simultaneous hostile stimuli.
Single cell surface engineering provides the most efficient, non-genetic strategy to enhance cell stability. However, it remains a huge challenge to improve cell stability in complex artificial environments. Here, a soft biohybrid interfacial layer is fabricated on individual living-cell surfaces by their exposure to a suspension of gold nanoparticles and l-cysteine to form a protecting functional layer to which porous silica layers were bound yielding pores with a diameter of 3.9 nm. The living cells within the bilayered nanoshells maintained high viability (96 ± 2%) as demonstrated by agar plating, even after five cycles of simultaneous exposure to high temperature (40 °C), lyticase and UV light. Moreover, yeast cells encapsulated in bilayered nanoshells were more recyclable than native cells due to nutrient storage in the shell.


Explanatory Models for Mental Distress Among University Students in Ethiopia: A Qualitative Study.
Socio-culturally determined processes account for how individuals give meanings to health, illness, causal attributions, expectations from treatment, and related outcomes. There is limited evidence of explanatory models for mental distress among higher education institutions in Ethiopia. The objective of this study was to explore the explanatory models for mental distress among Wolaita Sodo University. The current study used a phenomenological research approach, and we collected data from 21 students. The participants were purposively recruited based on eligibility criteria. Semi-structured interviews were conducted from December 2017 to January 2018 using the Short Explanatory Models Interview. The interviews were audio-recorded, transcribed into the Amharic language and translated into English. Data were analyzed using framework analysis with the assistance of open code software 4.02. Most students experienced symptoms of being anxious, fatigue, headaches and feelings of hopelessness. They labeled these symptoms like anxiety or stress. The most commonly reported causal explanations were psychosocial factors. Students perceived that their anxiety or stress was severe that mainly affected their mind, which in turn impacted their interactions with others, academic result, emotions and motivation to study. Almost all the students received care from informal sources, although they wanted to receive care from mental health professionals. They managed their mental distress using positive as well as negative coping strategies. The policy implication of our findings is that mental health interventions in higher education institutions in Ethiopia should take into account the explanatory models of students' psychological distress.


An autoinflammatory disease due to homozygous deletion of the IL1RN locus.
We describe a patient with an autoinflammatory disease in which the main clinical features are pustular rash, marked osteopenia, lytic bone lesions, respiratory insufficiency, and thrombosis. Genetic studies revealed a 175-kb homozygous deletion at chromosome 2q13, which encompasses several interleukin-1 family members, including the gene encoding the interleukin-1-receptor antagonist (IL1RN). Mononuclear cells, obtained from the patient and cultured, produced large amounts of inflammatory cytokines, with increasing amounts secreted after stimulation with lipopolysaccharide. A similar increase was not observed in peripheral-blood mononuclear cells from a patient with neonatal-onset multisystem inflammatory disorder (NOMID). Treatment with anakinra completely resolved the symptoms and lesions.


A case of mitral stenosis with achalasia.
A 65-year-old woman with rheumatic heart disease and severe mitral stenosis developed dysphagia. As her dysphagia could not be directly attributed to an enlarged left atrium, she underwent barium swallow, which established the diagnosis of achalasia. This case report shows two unrelated diseases present in the same patient which individually can cause dysphagia.


Simultaneous vitality and DNA-fragmentation measurement in spermatozoa of smokers and non-smokers.
Because cigarette smoke is a powerful ROS producer, we hypothesized that the spermatozoa of smokers would be more at risk of having increased DNA fragmentation than spermatozoa of non-smoking men. A cross-sectional study was performed on consenting smokers and non-smokers, consulting in an infertility clinic for routine sperm analysis. The application of a novel TUNEL assay coupled to a vitality marker, LIVE/DEAD®, allowed both DNA fragmentation and viability measurement within spermatozoa of participants to be analyzed by flow cytometry. The coupled vitality-DNA fragmentation analysis revealed that non-smokers and smokers, respectively presented medians of 3.6% [0.6-36.8] and 3.3% [0.9-9.6] DNA fragmented spermatozoa among the living spermatozoa population (P > 0.05). No deleterious effect of smoking on spermatozoa was found in our study. More studies concerning potential mutagenic capacities of cigarette smoke on spermatozoa are necessary. In addition, the coupled vitality-DNA fragmentation analysis may orient Assisted Reproductive Technology teams when confronted with patients having a high percentage of DNA-fragmented living spermatozoa.


Allele-specific late replication and fragility of the most active common fragile site, FRA3B.
FRA3B at 3p14.2 is the most active of the common fragile sites in the human genome and is expressed when cells are exposed to the DNA replication inhibitor, aphidicolin. Several lines of evidence suggest that fragile sites are regions of late replication. To elucidate the relationship between the timing of replication across the FRA3B region and its corresponding fragility, we labeled cells with 5-bromo-2'-deoxyuridine (BrdU) and adopted an immunofluorescent procedure to visualize late replicating DNA (BrdU-substituted DNA) in metaphase chromosomes. We also chose 21 markers along the FRA3B region and analyzed the timing of replication using BrdU-labeled DNA from different stages of the cell cycle sorted by flow cytometry. Our results show that there are two distinct alleles that replicate at different stages in the cell cycle and that breaks/gaps preferentially occurred on the chromosome 3 with the late replication allele. These results provide direct evidence that allele-specific late replication is involved in the fragility of the most active common fragile site, FRA3B.


Extensive C-terminal deletion in human immunodeficiency virus type 1 Env glycoprotein arising after long-term culture of chronically infected cells.
Human immunodeficiency virus type 1 (HIV-1) chronically infected (CI) cell lines were established from HIV-1HIB/LAI-infected MT-4 cells that survived acute infection. The HIV env gene expressed in the two long-term cultured cell lines differed from that of the lines cultured for shorter periods, by coding for a glycoprotein gp 160 that had the C terminus deleted. One long-term cultured cell line, CI-17, was studied in detail. An insertion of a premature stop codon in the env gene caused about 90% of gp160 molecules to be truncated (gp160x), lacking both cytoplasmic and transmembrane domains; these species were secreted into the cell medium, and could form oligomers with other truncated gp160 molecules as well as with their normal counterparts. CI-17 cells constantly yielded high levels of viral protein and relatively low quantities of infectious virus, without cytopathicity. However, acute infection of fresh MT-4 cells with CI-17-derived virus led to cytopathicity, the rate of which as well as the Env glycoprotein pattern depended on multiplicity: (i) using an infection dose of 10(-4) ID50/cell, cells died 7 to 8 days post-infection with normal gp160 synthesis predominating; (ii) with 10(-2) ID50, gp160x was produced as early as 48 h post-infection and cell death was delayed. Predominant gp160x formation occurred again when new CI cell lines were obtained with CI-17-derived virus. Thus, two human immunodeficiency virus variants, a normal and a defective one, are persistently expressed in CI-17 cells. The other long-term cultured CI cell line also expressed gp160 with a similar (albeit slightly longer) deletion of a C-terminal region in most molecules, but the cell lines that were cultured for shorter periods did not. These results suggest that the emergence of HIV variants with a C-terminal deletion in the Env glycoprotein, which coexist with normal virus, may play a role in maintaining the long-term growth capacity and viability of CI cells.


An investigation of support services available by Internet searching to families impacted by a parent's cancer.
Throughout a parent's cancer diagnosis and treatment, families are likely to turn to the Internet to find social, emotional, and informational support services. It is important that professionals understand the content and availability of support discoverable through Web searches. This study identified and described Web-based information regarding support services offered to families when a parent has cancer. Predefined key search terms were searched across 2 public Web browsers for a period of 3 months. Web-based information of support services (n = 25) was abstracted, and their key criteria examined. Of services offering face-to-face support (n = 22), more were founded by persons impacted by cancer (n = 10) (patients, parents, and offspring) than professionals (n = 4). Services offered online were created by health organisations and universities (n = 3). It appeared several service programs were delivered by volunteers, support staff, or facilitators, rather than registered health professionals or those with professional qualifications. Few services supported young children (under 5 years) or the parents themselves. Support services for families affected by a parent's cancer is relatively difficult to find through Web searches. Ambiguity around staff qualifications or evidence-base of support may hinder layperson engagement. Services offering face-to-face support were often founded by those directly affected by cancer; conversely, services founded by persons with assumable expertise (health organisation and university staff) were limited to online support. Such findings highlight a possible discrepancy in consumer and provider priorities and have implications for current understandings of psychosocial support needs and service development for families affected by a parent's cancer.


The synthesis of protected 5'-amino-2',5'-dideoxyribonucleoside-3'-O-phosphoramidites; applications of 5'-amino-oligodeoxyribonucleotides.
Synthetic routes to the four appropriately protected 5'-amino-2',5'-dideoxyribonucleoside-3'-O-(2-cyanoethyl N,N-diisopropylphosphoramidites) have been developed. The structures of all intermediates were confirmed by 13C n.m.r. spectroscopy. These building blocks have been used to prepare 5'-amino-oligodeoxyribonucleotides, which can be coupled to a wide variety of compounds, in particular metal cluster derivatives, but also fluorophores and biotin derivatives, thus generating a variety of very useful probes. Brief mention is made of a tetrairidium cluster derivative of 5'-amino-d[CCGATATCGG], which has been cocrystallised with EcoRV, and will be used for electron microscopy studies.


Continuous use of metformin in patients receiving contrast medium: what is the evidence? A systematic review and meta-analysis.
Substantial inconsistencies exist in current guidelines regarding recommendations of metformin usage with the administration of a contrast medium. We aimed to perform a meta-analysis to determine whether the risks of contrast-induced acute kidney injury (CI-AKI) and lactic acidosis increase with metformin use in diabetic patients receiving a contrast medium. Studies were retrieved from databases from inception to May 15, 2021. Studies that compared the outcomes of using metformin with not using metformin during contrast medium administration were included. The primary outcomes were incidence of CI-AKI and lactic acidosis. The secondary outcomes were renal function changes from baseline. Data analysis was using risk ratio (RR) for dichotomous outcomes and mean differences (MD) with 95% confidence intervals (CI) for continuous outcomes. Analyses of two randomized controlled trials and four retrospective cohorts examining a total of 1459 patients revealed no significant differences in the incidence of CI-AKI (RR = 1.08; 95% CI, 0.72 to 1.63) and in changes in renal function measurements (serum creatinine: MD = 0.00 mg/dL, 95% CI, - 0.05 to 0.05; estimated glomerular filtration rate: MD = 0.22, 95% CI, - 2.47 to 2.91) after contrast medium administration between patients using and not using metformin. There is no evidence that continuing metformin during contrast medium administration is associated with a higher risk of CI-AKI, lactic acidosis, or renal function deterioration compared to patients who discontinued metformin or who were not metformin users. The limited quality of the included studies may compromise the strength of evidence provided in this meta-analysis. There is no need to discontinue metformin either before or after intravenous contrast medium exposure in patients with eGFR > 30 mL/min/1.73 m2. In patients receiving intra-arterial contrast medium with first-pass renal exposure, there is no need to withhold metformin if eGFR is above 60 mL/min/1.73 m2. For patients who have an eGFR level between 30 and 60 mL/min/1.73 m2 and are receiving intra-arterial contrast medium with first-pass renal exposure, no case of lactic acidosis was observed based on present data, but further evidence is needed to make a strong suggestion regarding its safety.


Color-Kinematics Duality and Sudakov Form Factor at Five Loops for N=4 Supersymmetric Yang-Mills Theory.
Using color-kinematics duality, we construct for the first time the full integrand of the five-loop Sudakov form factor in N=4 super-Yang-Mills theory, including nonplanar contributions. This result also provides a first manifestation of the color-kinematics duality at five loops. The integrand is explicitly ultraviolet finite when D<26/5, coincident with the known finiteness bound for amplitudes. If the double-copy method could be applied to the form factor, this would indicate an interesting ultraviolet finiteness bound for N=8 supergravity at five loops. The result is also expected to provide an essential input for computing the five-loop nonplanar cusp anomalous dimension.


Some considerations in evaluating spoken word recognition by normal-hearing, noise-masked normal-hearing, and cochlear implant listeners. I: The effects of response format.
The purpose of the present studies was to assess the validity of using closed-set response formats to measure two cognitive processes essential for recognizing spoken words---perceptual normalization (the ability to accommodate acoustic-phonetic variability) and lexical discrimination (the ability to isolate words in the mental lexicon). In addition, the experiments were designed to examine the effects of response format on evaluation of these two abilities in normal-hearing (NH), noise-masked normal-hearing (NMNH), and cochlear implant (CI) subject populations. The speech recognition performance of NH, NMNH, and CI listeners was measured using both open- and closed-set response formats under a number of experimental conditions. To assess talker normalization abilities, identification scores for words produced by a single talker were compared with recognition performance for items produced by multiple talkers. To examine lexical discrimination, performance for words that are phonetically similar to many other words (hard words) was compared with scores for items with few phonetically similar competitors (easy words). Open-set word identification for all subjects was significantly poorer when stimuli were produced in lists with multiple talkers compared with conditions in which all of the words were spoken by a single talker. Open-set word recognition also was better for lexically easy compared with lexically hard words. Closed-set tests, in contrast, failed to reveal the effects of either talker variability or lexical difficulty even when the response alternatives provided were systematically selected to maximize confusability with target items. These findings suggest that, although closed-set tests may provide important information for clinical assessment of speech perception, they may not adequately evaluate a number of cognitive processes that are necessary for recognizing spoken words. The parallel results obtained across all subject groups indicate that NH, NMNH, and CI listeners engage similar perceptual operations to identify spoken words. Implications of these findings for the design of new test batteries that can provide comprehensive evaluations of the individual capacities needed for processing spoken language are discussed.


Enhanced ethylene separation and plasticization resistance in polymer membranes incorporating metal-organic framework nanocrystals.
The implementation of membrane-based separations in the petrochemical industry has the potential to reduce energy consumption significantly relative to conventional separation processes. Achieving this goal, however, requires the development of new membrane materials with greater selectivity, permeability and stability than available at present. Here, we report composite materials consisting of nanocrystals of metal-organic frameworks dispersed within a high-performance polyimide, which can exhibit enhanced selectivity for ethylene over ethane, greater ethylene permeability and improved membrane stability. Our results suggest that framework-polymer interactions reduce chain mobility of the polymer while simultaneously boosting membrane separation performance. The increased stability, or plasticization resistance, is expected to improve membrane utility under real process conditions for petrochemical separations and natural gas purification. Furthermore, this approach can be broadly applied to numerous polymers that encounter aggressive environments, potentially making gas separations possible that were previously inaccessible to membranes.


Current and emerging fixed combination therapies in glaucoma: a safety and tolerability review.
Fixed dose combinations (FCs) represent a potentially valuable treatment strategy in glaucoma management. Fixed combinations not only improve adherence by reducing the medication burden, but also decrease the total amount of potentially deleterious preservatives an eye is exposed to. We provide a critical review of selected evidence on both the safety and tolerability of presently available and emerging glaucoma FCs. There is convincing short-term safety and tolerability evidence on intraocular pressure (IOP)-lowering FCs compared to that of monotherapies and, to a lesser degree, to that of concomitant, equivalent combination therapies. In contrast, there is a scarcity of trials evaluating the long-term efficacy and safety of glaucoma FCs and no conclusive data on the reduction of adverse events with FCs. It is vital for clinicians to carefully weigh the efficacy, safety, tolerability, and adherence of IOP-lowering FCs. Given the number of currently available and emerging FC therapy options in glaucoma, as well as the complexities of incorporating them in the various combination therapy regimens, successful stepwise therapy remains often elusive.


Toward prediction of binding affinities between the MHC protein and its peptide ligands using quantitative structure-affinity relationship approach.
It is important but challenging to determine the binding specificity of MHC-peptide interactions accurately and to predict their binding affinity quantitatively. In this paper, we discuss the application of an effective amino acid descriptor to model and predict the binding affinities between the MHC protein and its peptide ligands. This amino acid descriptor was derived from 23 electronic properties, 37 steric properties, 54 hydrophobic properties and 5 hydrogen bond properties of coded amino acids using principal component analysis (PCA), called the divided physicochemical property scores (DPPS). The DPPS descriptor was used to characterize a set of mouse MHC (H-2K(K)) binding peptides, and genetic algorithm-partial least square (GA-PLS) models were then constructed. In analyses, these models were statistically consistent with previous reports and molecular graphics exhibition. Hydrophobic interactions and hydrogen bonds were important to antigen recognition and presentation, especially exerting effects on anchor residues of peptides.


Physiologically based biopharmaceutics modeling of regional and colon absorption in humans.
Colon absorption is a key determinant for successful development of extended release and colon targeted drug products. This is the first systematic evaluation of the ability to predict in vivo regional differences in absorption and the extent of colon absorption in humans using mechanistic physiologically based biopharmaceutics modeling (PBBM). A new dataset, consisting of 19 drugs with a wide range of biopharmaceutics properties and extent of colon absorption in humans, was established. Mechanistic predictions of the extent of absorption and plasma exposure after oral, or jejunal and direct colon administration were performed in GastroPlus and GI-Sim using an a priori approach. Two new colon models developed in GI-Sim, were also evaluated to assess if the prediction performance could be improved. Both GastroPlus and GI-Sim met the pre-defined criteria for accurate predictions of regional and colon absorption for high permeability drugs irrespective of formulation type, while the prediction performance was poor for low permeability drugs. For solutions, the two new GI-Sim colon models improved the colon absorption prediction performance for the low permeability drugs while maintaining the accurate prediction performance for the high permeability drugs. In contrast, the prediction performance decreased for non-solutions using the two new colon models. In conclusion, PBBM can be used with sufficient accuracy to predict regional and colon absorption in humans for high permeability drugs in candidate selection as well as early design and development of extended release or colon targeted drug products. The prediction performance of the current models needs to be improved to allow high accuracy predictions for commercial drug product applications including highly accurate predictions of the entire plasma concentration-time profiles as well as for low permeability drugs.


Crescentic lupus glomerulonephritis.
Crescentic lupus glomerulonephritis (greater than or equal to 50% crescents) occurred in 16% of systemic lupus erythematosus (SLE) patients biopsied over a 32-month period. All had underlying WHO class IV lupus nephritis. These patients more frequently manifested with acute renal failure usually of the non-oliguric type, had heavier proteinuria and lower serum albumin, but were otherwise indistinguishable from non-crescentic WHO class IV lupus nephritis in their other renal and extrarenal manifestations or in their serological findings. Crescentic lupus glomerulonephritis may occur in patients at first presentation with SLE, or develop in patients after prolonged follow-up initially for lupus nephritis of WHO class IV or other classes. Combined methylprednisolone pulse therapy, immunosuppressives, antiplatelet agents with or without anticoagulant produced good initial responses. Prognosis was unfavorable for inadequately treated patients or for patients with persistent nephrotic syndrome and crescents.


Early Experience with the HeartMate Percutaneous Heart Pump from the SHIELD II Trial.
The HeartMate Percutaneous Heart Pump (PHP) is a novel circulatory support catheter delivering a self-expanding 24 French impeller across the aortic valve. The SHIELD II trial compares outcomes among heart failure patients undergoing high-risk percutaneous coronary intervention (HR-PCI) with the PHP versus Impella systems. The trial was halted in 2017 due to device malfunctions. We aimed to describe procedural, hemodynamic, and clinical outcomes among HR-PCI patients treated with PHP as part of the SHIELD II trial roll-in phase. Procedural, hemodynamic, and 90 day outcomes were assessed among patients undergoing HR-PCI with a left ventricular ejection fraction ≤35% and last patent coronary conduit, unprotected left main disease, or significant three vessel disease. The primary endpoint was the 90 day composite of cardiovascular death, myocardial infarction, stroke, repeat revascularization, major bleeding, new/worsening aortic regurgitation, and severe hypotension. Among 75 roll-in phase patients, PHP support duration was 101 ± 53 minutes with 2.5 ± 1.4 coronary lesions treated per patient. Compared with predevice values, the PHP system increased cardiac power and mean arterial pressure. Maximum recorded device flows were 0.4-6.2 L/minute with 26% (n = 19/73) and 9.6% (n = 7/73) of patients achieving peak flows above 3.5 or 5.0 L/minute, respectively. Five PHP device malfunction events (6.7%) were observed. At 90 days, the composite endpoint occurred in 24.3% (18/74) of patients. Early PHP experience demonstrated successful device performance in the majority of enrolled patients; however, unexpected malfunctions led to device revision. Completion of the SHIELD II trial will be required to confirm the safety and efficacy of this iteration of the PHP system in HR-PCI.


[Profile of medical schools of the State of São Paulo, Brazil, in 1996].
The author examines different aspects of medical schools located in the region of São Paulo State (Brazil). There are 19 schools corresponding to 23% of the total number of medical schools in Brazil. The analysis includes elements about students and teachers, including their number and academic titulation; elements about teaching-learning process are also evaluated. An important contribution is the comparative analysis of the curricular evolution from 1986 to 1996.


Inhibition of Kaposi's sarcoma in vivo by fenretinide.
We examined the effects of fenretinide [N-(4-hydroxyphenyl)retinamide; (4HPR)] on highly angiogenic Kaposi's sarcoma tumors in vivo and investigated the mechanisms involved for potential clinical applications. (CD-1)BR nude mice bearing KS-Imm cell tumors were randomized to receive 4HPR or vehicle until sacrifice. In vitro, KS-Imm and endothelial cells were treated with 4HPR to study the effects on proliferation, apoptosis, migration, and invasion; in vivo angiogenesis was evaluated in the Matrigel model. Angiogenesis-related and retinoid receptor molecules were examined at the mRNA and protein expression levels. In vivo, 4HPR significantly (P<0.001) reduced growth of detectable Kaposi's sarcoma (KS) xenografts and inhibited angiogenesis in the Matrigel plug assay (P<0.04). In vitro, 4HPR affected KS-Imm and endothelial cell growth and KS-Imm migration and invasion. 4HPR invasion inhibition was associated with decreased release of matrix metalloprotease-2 and rapid reduction of vascular endothelial growth factor (VEGF) expression by KS cells and of vascular endothelial growth factor receptor 2 (VEGFR2) by KS and endothelial cells. Finally, 4HPR repression of angiogenesis was associated with a 4HPR-induced increase in retinoic acid receptor beta expression. These data indicate that 4HPR inhibits KS tumor growth in vivo through a mechanism involving the modulation of angiogenesis-associated growth factors and their receptors on both tumor and endothelial cells. In addition, 4HPR inhibited invasion by decreasing of matrix metalloprotease-2 activity. Our results justify further studies to evaluate the utility of 4HPR as a chemopreventive or therapeutic agent in KS, a malignancy associated with immune suppression that has a high risk of recurrence with highly active antiretroviral therapy failure.


Potential for Standardization and Automation for Pathology and Endoscopy in Inflammatory Bowel Disease.
Automated image analysis methods have shown potential for replicating expert interpretation of histology and endoscopy images, which traditionally require highly specialized and experienced reviewers. Inflammatory bowel disease (IBD) diagnosis, severity assessment, and treatment decision-making require multimodal expert data interpretation and integration, which could be significantly aided by applications of machine learning analyses. This review introduces fundamental concepts of machine learning for imaging analysis and highlights research and development of automated histology and endoscopy interpretation in IBD. Proof-of-concept studies strongly suggest that histologic and endoscopic images can be interpreted with similar accuracy as knowledge experts. Encouraging results support the potential of automating existing disease activity scoring instruments with high reproducibility, speed, and accessibility, therefore improving the standardization of IBD assessment. Though challenges surrounding ground truth definitions, technical barriers, and the need for extensive multicenter evaluation must be resolved before clinical implementation, automated image analysis is likely to both improve access to standardized IBD assessment and advance the fundamental concepts of how disease is measured.


Schizandrol A reverses multidrug resistance in resistant chronic myeloid leukemia cells K562/A02.
Overexpression of P-gp is the main cause of multidrug resistance (MDR) and chemotherapeutic failure in leukemia. In this study, the multidrug resistance reverse effect of Schizandrol A (SchA) was demonstrated with P-gp overexpressed drug-resistant K562/A02 cells. SchA had almost no cytotoxic activity, the EC50 value reversed to DOX was in the nanomole range of (707 ± 29nM) and had a high selectivity index (> 113) to normal cells. DOX accumulation and Rh123 efflux tests demonstrated that the MDR reversal activity of SchA was induced by inhibiting P-gp function. Western blotting assay showed that SchA down-regulated the expression of P-gp by inhibiting the PI3K / Akt signaling pathway, which was also a key factor in reversal activity. Therefore, SchA may be a potential candidate for natural MDR reversal agents.


Infection Caused by Phaeomoniella chlamydospora Associated with Esca-like Symptoms in Grapevine in Chile.
Trunk diseases such as esca have been recognized as an economically important problem of grapevine worldwide. A study was conducted to characterize the distribution of Phaeomoniella chlamydospora in Chile. A field survey of young and mature grapevines from 67 vineyards located along a 1,315-km north-south axis demonstrated that P. chlamydospora was present in 94.9% of the grapevine samples showing the black-wood streaking symptom (BWS) but not the characteristic foliar symptoms of esca. Phylogenetic analysis of the ribosomal DNA internal transcribed spacer (ITS) combined with β-tubulin (BT) genes grouped Chilean isolates together with reference isolates from South Africa and the United States, whereas Spanish isolates were clustered separately. Chilean isolates differed by only 2 to 3 bp for BT and ITS, respectively. Conidia germinated at 5 to 35°C, with an optimal temperature of 25 to 30°C. Isolates were pathogenic, and Koch's postulates were fulfilled in separate sets of inoculations of axenic plantlets, cuttings, 2-year-old plants, spurs, and shoots of V. vinifera. This study showed that P. chlamydospora was associated consistently with BWS and no other apparent symptom in young and mature grapevines, including nursery plants, in Chile. Inoculum was absent from the soil, grapevine pruning debris, sap samples, and herbaceous weeds, which is in contrast to past studies. At this time, Vitis spp. are the only known hosts of P. chlamydospora in Chile.


A Pragmatic, Single-Center, Prospective, Randomized Controlled Trial of Adjunct Hemoglobin-Mediated Granulox Topical Oxygen Therapy Twice Weekly for Foot Ulcers.
Achieving timely healing of foot ulcers can help avoid complications such as infection and amputation; topical oxygen therapy has shown promise in achieving this. We evaluated the clinical effectiveness of Granulox, a hemoglobin spray device designed to deliver oxygen to the surface of wounds, for the healing of foot ulcers. We conducted a single-center, prospective, randomized controlled trial comparing standard of care (once-weekly podiatric medical clinic visits) versus standard care plus adjunct Granulox therapy twice weekly in adults with foot ulcers. After a 2-week screening phase, patients in whom the index wound had healed by less than 50% were randomized 1:1. Outcome measures were collated during the trial phase at 6 and 12 weeks. Of 79 patients enrolled, 38 were randomized. After 12 weeks, the median percentage wound size reduction compared with the size of the ulcer at the start of the trial phase was 100% for the control arm and 48% for the Granulox arm (P = .21, Mann-Whitney U test). In the former, eight of 14 foot ulcers had healed; in the latter, four of 15 (P = .14, Fisher exact test). In the control arm, two amputations and one withdrawal occurred, whereas in the Granulox arm, one unrelated death and five withdrawals were recorded. We could not replicate the favorable healing associated with use of Granulox as published by others. Differences in wound chronicity and frequency of Granulox application might have influenced differences in study results. Granulox might perform best when used as an adjunct for treatment of chronic wounds at least 8 weeks old.


Confusional state in stroke: relation to preexisting dementia, patient characteristics, and outcome.
Acute confusional state (ACS) is frequent in hospitalized stroke patients. We previously showed that 16% of patients admitted for a stroke have preexisting dementia. The extent to which preexisting cognitive decline is associated with a risk of ACS at the acute stage of stroke remains to be systematically examined. The aim of this study was to evaluate the prevalence of ACS in acute stroke patients, to study the influence of preexisting cognitive decline and other patient characteristics, and to evaluate the influence of ACS on outcome. We diagnosed ACS using DSM-IV criteria and the Delirium Rating Scale with a cutoff of 10 in 202 consecutive stroke patients aged 40 years or older (median age, 75 years; range, 42 to 101 years). Cognitive functioning before stroke was assessed with the Informant Questionnaire on Cognitive Decline in the Elderly. Forty-nine stroke patients (24.3%; 95% CI, 18.3% to 30.2%) had an ACS during hospitalization. Using logistic regression analysis, we found preexisting cognitive decline (P=0.006) and metabolic or infectious disorders (P=0.008) to be independent predictors of ACS. Functional, but not vital, prognosis was worse in patients with ACS at discharge and 6 months after stroke. ACS occurs in one fourth of stroke patients older than 40 years. Its occurrence requires inquiry for a preexisting cognitive decline, which usually remains unrecognized in the absence of a systematic evaluation.


Characterization of exogenous bacterial oligosaccharyltransferases in Escherichia coli reveals the potential for O-linked protein glycosylation in Vibrio cholerae and Burkholderia thailandensis.
Bacterial protein glycosylation systems from varying species have been functionally reconstituted in Escherichia coli. Both N- and O-linked glycosylation pathways, in which the glycans are first assembled onto lipid carriers and subsequently transferred to acceptor proteins by an oligosaccharyltransferase (OTase), have been documented in bacteria. The identification and characterization of novel OTases with different properties may provide new tools for engineering glycoproteins of biotechnological interest. In the case of OTases involved in O-glycosylation (O-OTases), there is very low sequence homology between those from different bacterial species. The Wzy_C signature domain common to these enzymes is also present in WaaL ligases; enzymes involved in lipopolysaccharide biosynthesis. Therefore, the identification of O-OTases using solely bioinformatic methods is problematic. The hypothetical proteins BTH_I0650 from Burkholderia thailandensis E264 and VC0393 from Vibrio cholerae N16961 contain the Wzy_C domain. In this work, we demonstrate that both proteins have O-OTase activity and renamed them PglL(Bt) and PglL(Vc), respectively, similar to the Neisseria meningitidis counterpart (PglL(Nm)). In E. coli, PglL(Bt) and PglL(Vc) display relaxed glycan and protein specificity. However, effective glycosylation depends upon a specific combination of the protein acceptor, glycan and O-OTase analyzed. This knowledge has important implications in the design of glycoconjugates and provides novel tools for use in glycoengineering applications. The codification of enzymatically active O-OTase in the genomes of members of the Vibrio and Burkholderia genera suggests the presence of still unknown O-glycoproteins in these organisms, which might have a role in bacterial physiology or pathogenesis.


Tumor-associated macrophages promote angiogenesis and melanoma growth via adrenomedullin in a paracrine and autocrine manner.
Elevated numbers of tumor-associated macrophages (TAM) in the tumor microenvironment are often correlated with poor prognosis in melanoma. However, the mechanisms by which TAMs modulate melanoma growth are still poorly understood. This study was aimed at examining the function and mechanism of TAM-derived adrenomedullin (ADM) in angiogenesis and melanoma growth. We established in vitro and in vivo models to investigate the relationship between TAMs and ADM in melanoma, the role and mechanism of ADM in TAM-induced angiogenesis and melanoma growth. The clinical significance of ADM and its receptors was evaluated using melanoma tissue microarrays. ADM was expressed by infiltrating TAMs in human melanoma, and its secretion from macrophages was upregulated upon coculture with melanoma cells, or with melanoma cells conditioned media. Meanwhile, TAMs enhanced endothelial cell migration and tubule formation and also increased B16/F10 tumor growth. Neutralizing ADM antibody and ADM receptor antagonist, AMA, attenuated TAM-induced angiogenesis in vitro and melanoma growth in vivo, respectively. Furthermore, ADM promoted angiogenesis and melanoma growth via both the paracrine effect, mediated by the endothelial nitric oxide synthase signaling pathway, and the autocrine effect, which stimulated the polarization of macrophages toward an alternatively activated (M2) phenotype. Finally, immunofluorescence analysis on human melanomas showed that the expression of ADM in TAMs and its receptors was greatly increased compared with adjacent normal skins. Our study reveals a novel mechanism that TAMs enhance angiogenesis and melanoma growth via ADM and provides potential targets for melanoma therapies.


Evaluation of real-world persistence of propiverine and trospium chloride in treatment of overactive bladder in Germany.
Treatment persistence poses a crucial criterion for therapeutic success. Like in many other chronic diseases, in overactive bladder (OAB) syndrome also, many patients discontinue their treatment for diverse reasons. In order to evaluate the persistence in medication, this paper presents data of propiverine extended release (ER) and trospium chloride immediate release (IR) for three consecutive quarters, thus giving an insight into the probability of treatment discontinuation for these two drugs. Prescription data from German health insurances were analyzed. The frequency of follow-up prescriptions and drop-outs for propiverine ER and trospium chloride IR were compared for treatment-naïve, restarted, and switched patients after treatment initiation (quarter 1) for a period of 9 months (three consecutive calendar quarters). In all analyzed quarters, the percentage of follow-up prescriptions was significantly higher for propiverine ER than for trospium chloride IR. The chance of a follow-up prescription was also significantly higher for propiverine ER than for trospium chloride IR, although, in general, both drugs showed a decrease of follow-up prescriptions over time. This comparison of propiverine ER (once-daily administration) and trospium chloride IR (in a multiple dose administration) in patients with OAB syndrome demonstrates that there are substantial differences in the odds of a follow-up prescription. The longer a patient adheres to a therapeutic treatment plan, the better the chances for improvement of symptoms and having a positive impact on activities of daily living.


Elevated cerebrospinal fluid levels of monocyte chemotactic protein-1 correlate with HIV-1 encephalitis and local viral replication.
To investigate whether the CC-chemokine monocyte chemotactic protein (MCP)-1 could play a role in the pathogenesis of HIV infection of the central nervous system. This hypothesis was suggested by previous observations, including our finding of elevated cerebrospinal fluid (CSF) levels of this chemokine in patients with cytomegalovirus (CMV) encephalitis. CSF levels of MCP-1 were determined in 37 HIV-infected patients with neurological symptoms, and were compared with both the presence and severity of HIV-1 encephalitis at post-mortem examination and CSF HIV RNA levels. MCP-1 production by monocyte-derived macrophages was tested after in vitro infection of these cells by HIV. CSF MCP-1 levels were significantly higher in patients with (median, 4.99 ng/ml) than in those without (median, 1.72 ng/ml) HIV encephalitis. Elevated CSF MCP-1 concentrations were also found in patients with CMV encephalitis and with concomitant HIV and CMV encephalitis (median, 3.14 and 4.23 ng/ml, respectively). HIV encephalitis was strongly associated with high CSF MCP-1 levels (P = 0.002), which were also correlated to high HIV-1 RNA levels in the CSF (P = 0.007), but not to plasma viraemia. In vitro, productive HIV-1 infection of monocyte-derived macrophages upregulated the secretion of MCP-1. Taken together, these in vivo and in vitro findings support a model whereby HIV encephalitis is sustained by virus replication in microglial cells, a process amplified by recruitment of mononuclear cells via HIV-induced MCP-1.


Identification and estimation of microcystins in freshwater mussels.
Accumulation of microcystins mainly produced by cyanobacteria Microcystis was investigated for freshwater mussels and fishes collected from a lake where heavy blooms of Microcystis occurred every year. The identification of microcystins was performed by HPLC equipped with a frit FAB mass spectrometer. Microcystins LR and RR were identified in the mussels Unio douglasiae and Anadonta woodiana, whereas no microcystin was identified by the present method in fishes, such as Cyprinus carpio, Carassius carassius, and Hypomesius transpacificus.


Possible association between complex congenital heart defects and 11p15 hypomethylation in three patients with severe Silver-Russell syndrome.
Silver-Russell syndrome (SRS) is characterized by pre- and post-natal growth restriction that spares head growth, feeding difficulties, and variable dysmorphic facial features without major malformations. Hypomethylation of the paternal 11p15 imprinting control region 1 (ICR1) and maternal uniparental disomy of chromosome 7 are found in 50-60% and in 5-10% of SRS patients, respectively. We report on the pre- and post-natal features of three unrelated SRS patients with unusual congenital heart defects (CHDs). Two patients born prematurely had total anomalous pulmonary venous return and died shortly after birth, and a third patient, now 4 years old, had cor triatriatum sinistrum, which was surgically corrected. In all three patients, the underlying molecular defect was 11p15 ICR1 hypomethylation. Based on a large cohort with molecularly proven SRS, the prevalence of CHD in SRS is estimated at 5.5%. We suggest that the occurrence of CHD in SRS with 11p15 ICR1 hypomethylation is not coincidental, but specific to this genotype.


Structure, Process, and Outcomes of Liberian National Nursing and Midwifery Curricular Revisions.
The Republic of Liberia has had major disruptions to the education of its health care cadres. Post Ebola, the Resilient and Responsive Health Systems (RRHS) initiative began a new era of capacity building with the support of PEPFAR and HRSA. Nursing and Midwifery serve as the largest healthcare cadres in Liberia. The national nursing and midwifery curricula were overdue for the regulated review and revisions. The Science of Improvement was used as the framework to accomplish this multilateral activity. The Institute for Healthcare Improvement's (IHI) stages of improvement included: 1) Forming the team, 2) Setting the aims, 3) Establishing measures, 4) Selecting measures, 5) Testing changes, 6) Implementing changes, and 7) Spreading changes. These stages served as the blueprint for the structures and processes put into place to accomplish this national activity. The RN, Bridging, and BScM curricula all had redundant content that did not reflect teaching pedagogy and health priorities in Liberia. Courses were eliminated or reconfigured and new courses were created. Development of Nursing and Midwifery Curricular Taskforces were not as successful as was hoped. Two large stakeholder meetings ensured that this was the curricula of the Liberian faculty, deans and directors, and clinical partners. Monitoring and evaluation tools have been adopted by the Liberian Board for Nursing and Midwifery to serve as another improvement to check that the new curricula are being implemented and to identify gaps that may require future cycles of change for continued quality and improvement. Developing trust among the multilateral partners was critical to the success of this activity. Networks have been expanded, and a proposed pilot with the Ghana Board of Nursing and Midwifery and the US academic partner will examine the feasibility of implementing electronic licensing examinations for nurses and midwives.


Stapedectomy in children.
Stapes surgery for correction of conductive hearing loss in adults with otosclerosis is a well-established procedure. Its effectiveness in children, however, has received less scrutiny in the literature. Previous studies from our and other institutions demonstrated similar results in children and adults. Between 1980 and 1994 stapedectomies were done on 95 ears of 81 patients younger than 18 years (83 primary and 12 revisions). Data regarding age of onset, family history, associated anomalies, surgical findings, technique, hearing results, and complications were reviewed. Two groups were identified: congenital stapedial fixation and juvenile otosclerosis. Patients with congenital stapedial fixation had an earlier onset of hearing loss (3 vs 10 years, P < 0.001), a greater incidence of abnormalities of the malleus and incus (25% vs 3%, P < 0.001), and a slightly greater preoperative air-bone gap (35.2 +/- 12.9 vs 27.8 +/- 8.9, P = 0.002). Patients with otosclerosis had a greater frequency of a positive family history of deafness (53% vs 10%, P < 0.001). Overall, 79% of primary cases and 89% of revision cases had an improvement in hearing, with mean postoperative air-bone gaps of 15 dB and 22 dB, respectively. The gap did not widen significantly during the entire length of follow-up (mean 72 months). In primary cases, 59.1% obtained a postoperative air-bone gap of 10 dB or less. Eighty-two percent of children operated on for otosclerosis obtained excellent results (postoperative air-bone gap < or = 10 dB), compared with only 44% of children with congenital stapedial ankylosis (P = 0.02). In revision surgery, 29% of children obtained excellent results. Poorer results in both cases of congenital stapedial fixation and revision stapedectomy appear to be related to the greater incidence of associated anomalies of the malleus and incus. Indications, technique, complications, and considerations pertinent to childhood stapedectomy are discussed.


[The dermatologic spectrum of Lyme borreliosis].
The classical symptoms of Lyme borreliosis (LB) on the skin, erythema migrans, borrelia lymphocytoma and acrodermatitis chronica atrophicans (ACA) can be diagnosed clinically. In atypical cases, however, the diagnosis of a Borrelia burgdorferi (Bb) infection is based on the isolation of this bacterium from affected tissues. In early Lyme disease problems are arising in complicated erythema migrans which may represent reinfection, or with a failure of antibiotic treatment in some patients. The pathological changes in ACA presumably result from a chronic T-cell-mediated tissue injury with atrophy or sclerosis of connective tissue. On the other hand the presence of numerous plasma cells indicates activation of the humoral immune system where a progression into a malignant B-cell lymphoma is possible. In the recent past Bb could be cultivated from skin biopsies of a series of dermatoses of unknown origin, such as circumscribed scleroderma, dermatomyositis-like syndrome, relapsing nodular panniculitis, granuloma anulare and roseolar erythemas. New therapeutic strategies by administering antibiotics have been found to be effective in the described Bb induced dermatoses. Future studies should be focused on new standardized diagnostic procedures which make it possible to define the spectrum of dermatoses associated with a Bb infection.


Tolvaptan's Association with Low Risk of Acute Kidney Injury in Patients with Advanced Chronic Kidney Disease and Acute Decompensated Heart Failure.
Furosemide, a loop diuretic, is often empirically used to treat acute decompensated heart failure (ADHF) initially. Conversely, decongestion using tolvaptan, an aquaretic, is thought to maintain renal function compared to furosemide. However, it has not been investigated in patients with advanced chronic kidney disease (CKD) at high risk of developing acute kidney injury (AKI). This study aimed to investigate AKI incidence using tolvaptan add-on treatment, compared to increased furosemide treatment for patients with ADHF complicated by advanced CKD. We retrospectively studied patients with advanced CKD (estimated glomerular filtration rate [eGFR] <45 mL/min/1.73 m2) who developed ADHF under outpatient furosemide treatment. The exposure was set to tolvaptan add-on treatment, and the control was set to increased furosemide treatment. Of the 163 patients enrolled, 79 were in the tolvaptan group and 84 in the furosemide group. The mean age was 71.6 years, the percentage of males was 63.8%, the mean eGFR was 15.7 mL/min/1.73 m2, and patients with CKD stage G5 were 61.9%. AKI incidence was 17.7% in the tolvaptan group and 42.9% in the furosemide group (odds ratio [95% confidence interval]: 0.34 [0.13-0.86], p = 0.023 in multivariate logistic regression analysis). Persistent AKI incidence was 11.8% in the tolvaptan group and 32.9% in the furosemide group (odds ratio [95% confidence interval]: 0.34 [0.10-1.06], p = 0.066 in the multinomial logit analysis). This study suggests that tolvaptan may be better than furosemide in patients with ADHF experiencing complicated advanced CKD.


The complete mitochondrial genome and phylogenetic position of the critically endangered Trinidad Piping Guan, Pipile pipile synonym Aburria pipile (Aves: Galliformes).
The complete mitochondrial genome of the Critically Endangered Trinidad Piping Guan, Pipile pipile (Jacquin 1784) synonym Aburria pipile was sequenced for the first time in this study. The genome is 16,665 bp in length with overall base compositions of 30.1, 23.7, 32.3 and 13.9% for A, T, C, and G, respectively. Structurally, the P. pipile mitogenome is comparable to that of other Galliformes, thereby demonstrating typical avian gene organization. The mitogenome was subsequently used to produce a revised phylogenetic placement of P. pipile within the Galliforme order, positioning the Pipile genus basal within the Cracidae family. It is further envisaged that this novel genomic data will contribute to a wider understanding of genetic relationships within the genus Pipile and the analysis of the evolutionary relationships of the Galliforme order in a wider avian context.


Effects of adenosine receptor agonists and antagonists in a genetic animal model of primary paroxysmal dystonia.
1. Recent studies have shown beneficial effects of an adenosine A(2A) receptor agonist in dt(sz) mutant hamsters, an animal model of paroxysmal dystonia, in which stress and consumption of coffee can precipitate dystonic attacks. This prompted us to examine the effects of adenosine receptor agonists and antagonists on severity of dystonia in dt(sz) hamsters in more detail. 2. The non-selective adenosine A(1)/A(2A) receptor antagonists, caffeine (10 - 20 mg kg(-1) i.p.) and theophylline (10 - 30 mg kg(-1) s.c.), worsened the dystonia in dt(sz) hamsters. 3. Aggravation of dystonia was also caused by the selective adenosine A(1)/A(2A) antagonist CGS 15943 (9-chloro2-2-furyl)[1,2,4]triazolo[1,5-c]quinazolin-5-amine) at a dose of 30 mg kg(-1) i.p. and by the adenosine A(1) antagonist DPCPX (8-cyclopentyl-1,3-dipropylxanthine; 20 - 30 mg kg(-1) i.p.), while the A(2) antagonist DMPX (3,7-dimethyl-1-propargylxanthine; 2 - 4 mg kg(-1) i.p.) and the highly selective A(2A) antagonist ZM 241385 (4-(2-[7-amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol; 2 - 5 mg kg(-1) i.p.) failed to exert any effects on dystonia. 4. In contrast to the antagonists, both the adenosine A(1) receptor agonist CPA (N(6)-cyclopentyladenosine; 0.1 - 1.0 mg kg(-1) i.p.) and the A(2A) agonist CGS 21680 (2p-(2carboxyethylphen-ethylamino-5'-N-ethylcarboxamindoadenosine; 0.1 - 2.0 mg kg(-1) i.p.) exerted a striking improvement of dystonia. 5. These data suggest that the precipitating effects of methylxanthines are, at least in part, related to their adenosine receptor antagonistic action. 6. Although adenosine receptor agonists can be regarded as interesting candidates for the therapy of paroxysmal dystonia, adverse effects may limit the therapeutic potential of adenosine A(1) agonists, while beneficial effects of the adenosine A(2A) agonist CGS 21680 were already found at well tolerated doses.


Analysis of rare thalassemia caused by HS-40 regulatory site deletion.
ABSTRACT Objectives: To investigate the effect of HS-40 regulatory site deletion on α-globin gene expression and its clinical significance. Methods: Venous blood samples of subjects were analyzed using a hematology analyzer and high- performance liquid chromatography; fetal cord blood was analyzed by a capillary electrophoresis analyzer. Gap-polymerase chain reaction (PCR), reverse dot blot (RDB), and multiple-link-dependent probe amplification (MLPA) were used for genotyping of thalassemia. Results: The proband was POLR3 K, HS-40 heterozygous deletion; the proband's wife was -SEA/αα; the fetus was POLR3 K, HS-40 heterozygous deletion combined with -SEA deletion; all of them had microcytic hypochromic anemia. Fetal umbilical cord blood electrophoresis revealed a suspected Hb Bart's band to be 88.4%, and the fetus was, thus, diagnosed as Hb Bart's fetus. The red blood cell parameters of the sporadic case showed that he had microcytic hypochromic anemia. Hemoglobin (Hb) electrophoresis analysis showed Hb H to be 5.3%, leading to a diagnosis of Hb H disease. Gap-PCR and RDB identified the genotype to be -α3.7/αα, βA/βA. MLPA detected heterozygous deletion or -α3.7 deletion on one allele and deletion of the HS-40 regulatory site on the other allele. Conclusion: The deletion of HS-40 regulatory site reduced expression of α-globin. HS-40 heterozygous deletion manifested as mild anemia, which was of microcytic hypochromic type. When compounded with -α3.7/αα, it manifested as Hb H disease; and when compounded with -SEA/αɑ, it manifested as Hb Bart's fetus.


Scientific evaluation of spinal implants: an ethical necessity.
The clinical introduction of novel medical devices often occurs without evidence of good methodological quality and with relatively little oversight and regulation. As a consequence, the safety, efficacy, and long-term effects of devices are frequently insufficiently known upon device approval. Recent controversies surrounding the Poly Implant Prothèse (PIP) breast implants, metal-on-metal hip implants, and interspinous implants underscore the need to reconsider how innovation in medical devices can adhere to sound ethical standards without inhibiting surgical research and development. In this article, the introduction of spinal implants is taken as an example to firstly discuss the scientific and ethical challenges of developing, testing, and introducing novel medical devices and to secondly identify avenues for improving the existing regulatory frameworks for such innovation. Two measures for improvement are most feasible in the short term: demanding prospective studies before device introduction and developing registries to monitor and evaluate new medical devices. Level of evidence: 5.


Morphological appearance manifolds in computational anatomy: groupwise registration and morphological analysis.
Existing approaches to computational anatomy assume that a perfectly conforming diffeomorphism applied to an anatomy of interest captures its morphological characteristics relative to a template. However, the amount of biological variability in a groupwise analysis renders this task practically impossible, due to the nonexistence of a single template that matches all anatomies in an ensemble, even if such a template is constructed by group averaging procedures. Consequently, anatomical characteristics not captured by the transformation, and which are left out in the residual image, are lost permanently from subsequent analysis, if only properties of the transformation are examined. This paper extends our recent work [Makrogiannis, S., Verma, R., Davatzikos, C., 2007. Anatomical equivalence class: a computational anatomy framework using a lossless shape descriptor. IEEE Trans. Biomed. Imag. 26(4), 619-631] on characterizing subtle morphological variations via a lossless morphological descriptor that takes the residual into account along with the transformation. Since there are infinitely many [transformation, residual] pairs that reconstruct a given anatomy, we treat them as members of an Anatomical Equivalence Class (AEC), thereby forming a manifold embedded in the space spanned by [transformation, residual]. This paper develops a unique and optimal representation of each anatomy that is estimated from the corresponding AECs by solving a global optimization problem. This effectively determines the optimal template and transformation parameters for each individual anatomy, and eliminates respective confounding variation in the data. It, therefore, constitutes the second novelty, in that it represents a group-wise optimal registration strategy that individually adjusts the template and the smoothness of the transformation according to each anatomy. Experimental results support the superiority of our morphological analysis framework over conventional analysis, and demonstrate better diagnostic accuracy.


Congenital cardiac abnormalities in monozygotic twins. Report and review of the literature.
A pair of monozygotic twin girls is reported with concordance for 3 congenital cardiac abnormalities: (1) secundum atrial septal defect, (2) aneurysm of the membraneous ventricular septum, and (3) electrocardiographic frontal plane left axis deviation. A review of the published materials shows a 9.5 per cent incidence of concordance for congenital heart disease among monozygotic twins. In those in whom a precise cardiological diagnosis was made, 15/16 pairs (95%) were concordant for a specific defect, 2 had an additional defect, and only 1 pair had completely dissimilar defects. Concordance for congenital heart disease in monozygotic twins is uncommon, but when it occurs the defects will most often be identical.


Two major forms of DNA (cytosine-5) methyltransferase in human somatic tissues.
Thus far, only one major form of vertebrate DNA (cytosine-5) methyltransferase (CpG MTase, EC 22.214.171.124) has been identified, cloned, and extensively studied. This enzyme, dnmt1, has been hypothesized to be responsible for most of the maintenance as well as the de novo methylation activities occurring in the somatic cells of vertebrates. We now report the discovery of another abundant species of CpG MTase in various types of human cell lines and somatic tissues. Interestingly, the mRNA encoding this CpG MTase results from alternative splicing of the primary transcript from the Dnmt1 gene, which incorporates in-frame an additional 48 nt between exons 4 and 5. Furthermore, this 48-nt exon sequence is derived from the first, or the most upstream, copy of a set of seven different Alu repeats located in intron 4. The ratios of expression of this mRNA to the expression of the previously known, shorter Dnmt1 mRNA species, as estimated by semiquantitative reverse transcription-PCR analysis, range from two-thirds to three-sevenths. This alternative splicing scheme of the Dnmt1 transcript seems to be conserved in the higher primates. We suggest that the originally described and the recently discovered forms of CpG MTase be named dnmt1-a and dnmt1-b, respectively. The evolutionary and biological implications of this finding are discussed in relation to the cellular functions of the CpG residues and the CpG MTases.


Two cases of subacute sclerosing panencephalitis associated with brainstem involvement.
The most commonly involved areas in subacute sclerosing panencephalitis (SSPE) are periventricular and subcortical white matter. The basal ganglia, cerebellum, spinal cord and corpus callosum are less commonly involved. Brainstem involvement is rare and usually accompanied by other intracranial lesions. In this article, we report two cases of SSPE associated with brainstem involvement. The first case a 9-year-old girl had the typical symptom of SSPE. Magnetic resonance imaging (MRI) of the brain revealed a focal lesion 2 x 2.5 cm a diameter in the pons. The second case was a 6-year-old girl. On admission, MRI of the brain was normal. During 6th month of follow-up, T2-weighted MRI showed a hyperintense lesion in the pons and pedincule of cerebellum. On account of these cases we would like to stress that brainstem involvement may be seen in patients with SSPE; therefore, these patients should be monitored for this disorder.


Ocean tide and waves beneath the ross ice shelf, antarctica.
The ocean tide in the southern Ross Sea is principally diurnal. The tropic tide range (double amplitude) is between 1 and 2 meters, depending on the location, and is closely related to the local water-layer thickness. The range of the tropic tide is more than three times the range of the equatorial tide. Cotidal and coamplitude charts were made for the largest diurnal constituents, K(1) and O(1) and a provisional cotidal map was made for the semidiurnal constituent M(2). The amplitudes of the diurnal tide constituents are larger in the Ross Sea than in the adjacent southern Pacific Ocean, indicating the existence of a diurnal resonance related to the shape and depth of the sea. Waves related to ocean swell propagate into the ice-covered region from the northern Ross Sea. These waves have amplitudes near 1 centimeter, and periods in the range 1 to 15 minutes. The speed at which these waves travel is successfully predicted by flexural wave theory.


Identification of in vitro electropermeabilization equivalent pulse protocols.
Exposure of cells to an external sufficiently strong electric field results in the formation of pores across the membrane. This phenomenon, termed electropermeabilization, permits the transport of poorly permeant molecules into cytosol. In clinical practice, cell membrane permeabilization for drug electrotransfer is achieved using the ESOPE pulse protocol (1000 V/cm, 8 pulses, 100 μs, 5 kHz). The aim of this study was to investigate several combinations of electric field amplitude and pulse number able to induce electropermeabilization as the one observed when the ESOPE protocol was applied. Decreasing electric field amplitudes (1000 to 300 V/cm) in combination with increasing number of pulses (8 to 320) were applied to in vitro MG63 cells. Propidium iodide and Calcein blue AM uptake were used to evaluate cell electropermeabilization and viability. Results showed that the threshold of local electric field needed to obtain electropermeabilization decreased exponentially with increasing the number of pulses delivered (r2 5 0.92, p<0.0001). The absorbed dose threshold was dependent on the number of pulses for each voltage applied (r2 5 0.96, p<0.0001). In conclusion, the possibility of applying an increased number of pulses rather than increasing the electric field amplitude to perform electropermeabilization, may become an important tool for electropermeabilization-related clinical applications.


Effects of garlic powder supplementation on insulin resistance, oxidative stress, and body composition in patients with non-alcoholic fatty liver disease: A randomized controlled clinical trial.
Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide. Insulin resistance, oxidative stress, and obesity are major contributors to NAFLD pathogenesis. The effects of garlic powder supplementation on these risk factors in patients with NAFLD was investigated. In this 12-wk, randomized controlled clinical trial, ninety patients with NAFLD were randomly assigned to two groups. The treatment group received four tablets of garlic (each coated tablet contained 400 mg garlic powder) daily and the control group received four tablets of placebo (each coated tablet contained 400 mg starch). A significant decrease was seen in the treatment group compared to the control group in waist circumference (P = 0.001), body fat percent (P < 0.001), serum concentration of fasting blood sugar (P = 0.01), insulin (P < 0.001), homeostatic model assessment for insulin resistance (P < 0.001), and malondialdehyde (P < 0.001), as well as significant increase in skeletal muscle mass (P = 0.002), serum concentration of superoxide dismutase (P < 0.001), and total antioxidant capacity (P < 0.001). Garlic powder supplementation improved risk factors of NAFLD. Further studies are needed to determine the effects of garlic on hepatic features in patients with NAFLD. The study protocol was registered at Iranian clinical trials website under code IRCT20170206032417N4.


Induction of fos-like immunoreactivity in hypothalamic corticotropin-releasing factor neurons after adrenalectomy in the rat.
To identify brain sites responding to the removal of corticosterone feedback by adrenalectomy (ADX), rat brains were processed for fos immunocytochemistry 1, 3, and 7 days after ADX, sham-ADX, or no surgery using a polyclonal antiserum to fos residues 132-154. Compared to SHAM, ADX rats exhibited strong fos-like immunoreactivity (FLI) only in the parvocellular neurons of the paraventricular hypothalamic nuclei (PVN) 1, 3, and 7 days after surgery. Replacement with a corticosterone pellet at the time of adrenalectomy (ADX + B) prevented this increase in PVN FLI in three of four rats at 1 day, all rats at 3 days, and two of seven rats 7 days after surgery; 100 micrograms/ml corticosterone in the drinking water for 2 days before perfusion reversed ADX-induced increases in PVN FLI in 7-day ADX rats. Providing 25 micrograms/ml corticosterone in the drinking water to ADX rats for 5 days after surgery did not prevent expression of PVN FLI, even though this dose has been shown to normalize morning basal ACTH levels in ADX rats. Virtually all parvocellular PVN neurons expressing FLI after ADX costained for CRF. Some parvocellular neurons also expressed both fos and vasopressin. In all rats, many brain regions expressed FLI that was not related to adrenalectomy. We conclude that the changes in neuronal FLI correlate with demonstrated changes in neuroendocrine activity after ADX; however, suppression of ADX-induced FLI may require higher replacement levels of corticosterone than inhibition of ADX-induced ACTH secretion.