[A promoter responsible for over-expression of cholera toxin B subunit in cholera toxin A subunit structure gene].
A promoter sequence, which promotes the transcription of cholera toxin B subunit gene, was found in cholera toxin A subunit structure gene. The transcription starts at the adenine Located at +833, that is 456bp upstream to the A of the initiation codon ATG of cholera toxin B gene. Under the control of the promoter, cholera toxin B subunit was over-expressed as high as 200 mg/L at an optimized culture condition. The chloramphenicol acetyl transferase gene and beta-galactosidase could also be efficiently expressed under the direction of the promoter. This promoter may be responsible for the 6 fold and 7 fold higher expression level of cholera toxin B subunit than cholera toxin A subunit in V. cholerae and Escheria coli respectively. The over-expression of CTB may be useful in preparing vaccine against cholera and facilitating the construction of peptide-bearing immunogenic hybrid proteins.


Urban farming and malaria risk factors in a medium-sized town in Cote d'Ivoire.
Urbanization occurs at a rapid pace across Africa and Asia and affects people's health and well-being. A typical feature in urban settings of Africa is the maintenance of traditional livelihoods, including agriculture. The purpose of this study was to investigate malaria risk factors in urban farming communities in a medium-sized town in Côte d'Ivoire. Two cross-sectional surveys were carried out among 112 households from six agricultural zones. First, the heads of households were interviewed on agricultural land use, farming practices, water storage, sanitation facilities, and socioeconomic status. Second, a finger prick blood sample was taken from all household members and examined for the occurrence and density of Plasmodia. Geographic coordinates of houses, farming plots, and potential mosquito breeding sites were recorded and integrated into a geographic information system. Predictors of Plasmodium falciparum parasitemia were assessed using non-random and random effects Bayesian regression models. The overall prevalence of P. falciparum was 32.1%. In children < 15 years of age, risk factors for a P. falciparum infection included living in a specific agricultural zone, close proximity to permanent ponds and fish ponds, periodic stays overnight in temporary farm huts, and low socioeconomic status. Our findings indicate that specific crop systems and specific agricultural practices may increase the risk of malaria in urban settings of tropical Africa.


The siRNA sequence and guide strand overhangs are determinants of in vivo duration of silencing.
The use of short interfering RNAs (siRNA) in animals for target validation or as potential therapeutics is hindered by the short physical half-life when delivered as unencapsulated material and in turn the short active half-life of siRNAs in vivo. Here we demonstrate that the character of the two 3'-overhang nucleotides of the guide strand of siRNAs is a determinant of the duration of silencing by siRNAs both in vivo and in tissue culture cells. We demonstrate that deoxyribonucleotides in the guide strand overhang of siRNAs have a negative impact on maintenance of both the in vitro and in vivo activity of siRNAs over time. Overhangs that contain ribonucleotides or 2'-O-methyl modified nucleotides do not demonstrate this same impairment. We also demonstrate that the sequence of an siRNA is a determinant of the duration of silencing of siRNAs directed against the same target even when those siRNAs have equivalent activities in vitro. Our experiments have determined that a measurable duration parameter exists, distinct from both maximum silencing ability and the potency of siRNAs. Our findings provide information on incorporating chemically modified nucleotides into siRNAs for potent, durable therapeutics and also inform on methods used to select siRNAs for therapeutic and research purposes.


Nickel-Catalyzed Reductive Cross-Coupling of Aryl Bromides with Alkyl Bromides: Et3N as the Terminal Reductant.
Reductive cross-coupling has emerged as a direct method for the construction of carbon-carbon bonds. Most cobalt-, nickel-, and palladium-catalyzed reductive cross-coupling reactions to date are limited to stoichiometric Mn(0) or Zn(0) as the reductant. One nickel-catalyzed cross-coupling paradigm using Et3N as the terminal reductant is reported. By using this photoredox catalysis and nickel catalysis approach, a direct Csp(2)-Csp(3) reductive cross-coupling of aryl bromides with alkyl bromides is achieved under mild conditions without stoichiometric metal reductants.


In vitro selection of bispecific diabody fragments using covalent bicistronic DNA display.
Bispecific antibodies with two different antigen-binding sites have been widely used for a variety of medical applications. The activity and stability of antibody fragments can be improved by in vitro evolution. Although the affinity and stability of small bispecific antibody fragments such as diabodies can be further optimized by in vitro display technologies, cell-free display of bispecific antibody fragments has not been reported. In this study, we applied a covalent bicistronic DNA display for the in vitro selection of heterodimeric diabodies. First, we confirmed the antigen-binding activities of a diabody synthesized by an in vitro transcription and translation system. However, when we performed DNA-display selection of a model diabody library in a proof-of-principle experiment, no enrichment of the diabody gene was observed, likely due to a low yield of the diabody heterodimer. To overcome this issue, we introduced cysteine residues at the VH-VL interface of the diabody heterodimer. Using the disulfide-stabilized diabodies, we successfully enriched the diabody gene from a model library. Our results indicate that the covalent bicistronic DNA display technique could be useful for improving the stability and affinity of bispecific diabody fragments.


Cytosolic calcium measurements in renal epithelial cells by flow cytometry.
A variety of cellular processes, both physiological and pathophysiological, require or are governed by calcium, including exocytosis, mitochondrial function, cell death, cell metabolism and cell migration to name but a few. Cytosolic calcium is normally maintained at low nanomolar concentrations; rather it is found in high micromolar to millimolar concentrations in the endoplasmic reticulum, mitochondrial matrix and the extracellular compartment. Upon stimulation, a transient increase in cytosolic calcium serves to signal downstream events. Detecting changes in cytosolic calcium is normally performed using a live cell imaging set up with calcium binding dyes that exhibit either an increase in fluorescence intensity or a shift in the emission wavelength upon calcium binding. However, a live cell imaging set up is not freely accessible to all researchers. Alternative detection methods have been optimized for immunological cells with flow cytometry and for non-immunological adherent cells with a fluorescence microplate reader. Here, we describe an optimized, simple method for detecting changes in epithelial cells with flow cytometry using a single wavelength calcium binding dye. Adherent renal proximal tubule epithelial cells, which are normally difficult to load with dyes, were loaded with a fluorescent cell permeable calcium binding dye in the presence of probenecid, brought into suspension and calcium signals were monitored before and after addition of thapsigargin, tunicamycin and ionomycin.


Atypical fibroxanthoma.
This is the first case report of atypical fibroxanthoma in the otolaryngology literature. Atypical fibroxanthoma is a rare fibrohistiocytic tumor that occurs on actinically damaged skin. The typical presentation is a nodular lesion in sun-exposed areas of the head and neck in an elderly patient. Differentiating atypical fibroxanthoma from other soft tissue sarcomas, such as dermatofibrosarcoma protuberans and malignant fibrous histiocytoma, can be difficult. Treatment should be wide local excision with a 1-cm margin. In cosmetically important regions in which tissue preservation is important, Mohs' surgery should be considered.


An update on biomarker discovery and use in systemic sclerosis.
Systemic sclerosis (SSc) is an autoimmune disease characterized by excessive extracellular matrix deposition in the skin and internal organs. Three major abnormalities, autoimmunity, vasculopathy, and fibrosis, are considered to play important roles in the pathophysiology of SSc. SSc is a heterogeneous disease with clinical features, disease progress, therapeutic response, and prognosis. Therefore, identification of biomarkers, which can predict the course of the disease, is required for the progress of clinical practice. Areas covered: This review focuses on various SSc biomarkers for diagnosis, disease severity, and activity, as well as newly emerging and promising SSc biomarkers. Search was through PubMed and we focus on the papers in the last 5 years. Expert commentary: Biomarkers for SSc can be categorized into several groups: activity biomarker, severity biomarker, predictive biomarker, and biomarkers for specific clinical features (skin fibrosis, lung fibrosis, pulmonary arterial hypertension, peripheral vasculopathy, gastrointestinal, and malignancy). Some chemokines, such as CCL2 and CXCL4, have been identified as biomarkers of skin and lung fibrosis in SSc. In addition, anti-RNA polymerase III antibody has been noted to be a predictive biomarker of gastric antral vascular ectasia and malignancy.


Neglected issues concerning teaching human adrenal steroidogenesis in popular biochemistry textbooks.
In the human body, the adrenal steroids collectively regulate a plethora of fundamental functions, including electrolyte and water balance, blood pressure, stress response, intermediary metabolism, inflammation, and immunity. Therefore, adrenal steroidogenesis is an important biochemistry topic for students to learn in order for them to understand health consequences caused by deficiencies of enzymes in the adrenal steroidogenic pathways. However, popular biochemistry textbooks contain insufficient information and may sometimes give students a misimpression about certain aspects of human adrenal steroidogenesis. This article highlights two neglected issues in teaching human adrenal steroidogenesis in popular biochemistry textbooks. The purpose of this article is to draw attention to these issues. © 2017 by The International Union of Biochemistry and Molecular Biology, 45(6):469-474, 2017.


Melatonin receptor genes.
Melatonin is produced rhythmically by the pineal gland and the retina with increased synthesis during darkness. Pineal melatonin serves as the 'chemical expression of darkness' conveying information on the ambient light-dark cycle into rhythmic bodily functions. On-going debate on modes and sites of action ranges from views of melatonin affecting each and every cell ('cure-all') to those of melatonin having restricted actions through specific high-affinity receptors. The present review deals with the latter view. The use of 2-[125I]-iodomelatonin has allowed the exact localization and characterization of high-affinity melatonin receptors that signal through the G(i/o) class of G proteins. Molecular cloning of melatonin receptor genes has confirmed that most, if not all, high-affinity melatonin-binding sites represent the G-protein-coupled melatonin receptors. Based on sequence dissimilarities, melatonin receptors are classified into three subtypes, Mel1a, Mel1b and Mel1c. A distribution wider than originally thought of melatonin receptors in the human brain and peripheral sites has brought these receptors into focus of several drug companies, promising exciting times for research on melatonin and new therapeutic possibilities.


Functional group diversity of bee pollinators increases crop yield.
Niche complementarity is a commonly invoked mechanism underlying the positive relationship between biodiversity and ecosystem functioning, but little empirical evidence exists for complementarity among pollinator species. This study related differences in three functional traits of pollinating bees (flower height preference, daily time of flower visitation and within-flower behaviour) to the seed set of the obligate cross-pollinated pumpkin Cucurbita moschata Duch. ex Poir. across a land-use intensity gradient from tropical rainforest and agroforests to grassland in Indonesia. Bee richness and abundance changed with habitat variables and we used this natural variation to test whether complementary resource use by the diverse pollinator community enhanced final yield. We found that pollinator diversity, but not abundance, was positively related to seed set of pumpkins. Bees showed species-specific spatial and temporal variation in flower visitation traits and within-flower behaviour, allowing for classification into functional guilds. Diversity of functional groups explained even more of the variance in seed set (r2=45%) than did species richness (r2=32%) highlighting the role of functional complementarity. Even though we do not provide experimental, but rather correlative evidence, we can link spatial and temporal complementarity in highly diverse pollinator communities to pollination success in the field, leading to enhanced crop yield without any managed honeybees.


Human papillomavirus-negative epithelial proliferations resembling condylomata acuminata in a patient receiving vemurafenib for Stage IV melanoma.
With the discovery of v-Raf murine sarcoma viral oncogene homolog B (BRAF) inhibitors, new treatment possibilities arose against metastatic melanoma. A frequent adverse effect of BRAF inhibitor therapy is the induction of epithelial proliferations such as cutaneous squamous cell carcinoma and verrucous papilloma. Here, we describe a case in which a patient developed extensive anal epithelial proliferations resembling condylomata acuminata, after starting vemurafenib treatment. This adverse effect has rarely been reported in the literature. Interestingly, the lesions in our patient were negative for human papillomavirus, and mutations in BRAF, Neuroblastoma rat sarcoma viral oncogene homolog (NRAS), Kirsten rat sarcoma viral oncogene homolog (KRAS), and Harvey rat sarcoma viral oncogene homolog (HRAS) were not detected. Different pathways can contribute to these epithelial proliferations resembling condylomata acuminata. We show the relevance of a detailed history at the beginning and during treatment, instructions, education, and dermatological follow-up (including the genital area) for patients treated with BRAF inhibitors. Condylomata acuminata can influence the quality of life and are treated, in an early stage, with cryotherapy, coagulation, imiquimod, and/or CO2 laser therapy.


From Si(II) to Si(IV) and Back: Reversible Intramolecular Carbon-Carbon Bond Activation by an Acyclic Iminosilylene.
Reversibility is fundamental for transition metal catalysis, but equally for main group chemistry and especially low-valent silicon compounds, the interplay between oxidative addition and reductive elimination is key for a potential catalytic cycle. Herein, we report a highly reactive acyclic iminosilylsilylene 1, which readily performs an intramolecular insertion into a C═C bond of its aromatic ligand framework to give silacycloheptatriene (silepin) 2. UV-vis studies of this Si(IV) compound indicated a facile transformation back to Si(II) at elevated temperatures, further supported by density functional theory calculations and experimentally demonstrated by isolation of a silylene-borane adduct 3 following addition of B(C6F5)3. This tendency to undergo reductive elimination was exploited in the investigation of silepin 2 as a synthetic equivalent of silylene in the activation of small molecules. In fact, the first monomeric, four-coordinate silicon carbonate complex 4 was isolated and fully characterized in the reaction with carbon dioxide under mild conditions. Additionally, the exposure of 2 to ethylene or molecular hydrogen gave silirane 5 and Si(IV) dihydride 6, respectively.


Solvent-Driven Dynamical Crossover in the Phenylalanine Side-Chain from the Hydrophobic Core of Amyloid Fibrils Detected by 2H NMR Relaxation.
Aromatic residues are important markers of dynamical changes in proteins' hydrophobic cores. In this work we investigated the dynamics of the F19 side-chain in the core of amyloid fibrils across a wide temperature range of 300 to 140 K. We utilized solid-state 2H NMR relaxation to demonstrate the presence of a solvent-driven dynamical crossover between different motional regimes, often also referred to as the dynamical transition. In particular, the dynamics are dominated by small-angle fluctuations at low temperatures and by π-flips of the aromatic ring at high temperatures. The crossover temperature is more than 43 degrees lower for the hydrated state of the fibrils compared to the dry state, indicating that interactions with water facilitate π-flips. Further, crossover temperatures are shown to be very sensitive to polymorphic states of the fibrils, such as the 2-fold and 3-fold symmetric morphologies of the wild-type protein as well as D23N mutant protofibrils. We speculate that these differences can be attributed, at least partially, to enhanced interactions with water in the 3-fold polymorph, which has been shown to have a water-accessible cavity. Combined with previous studies of methyl group dynamics, the results highlight the presence of multiple dynamics modes in the core of the fibrils, which was originally believed to be quite rigid.


In-vitro and clinical evaluation of an oral mucosal adhesive film containing indomethacin.
To develop a new mucoadhesive film containing an analgesic combining clinical efficacy and patient comfort, we prepared and evaluated a two-layered film consisting of an adhesive layer containing indomethacin (IM) as the active ingredient and carboxyvinyl polymer (CP) as a bonding agent and a nonadhesive layer containing polyethylene glycol (PEG) to improve film texture. In in vitro and in vivo adhesive tests, the optimal concentration of CP that could be applied to the mucous membrane was 0.2% or 0.3%. Stability testing determined that the optimal storage conditions and expiration period were 4 degrees C without shade and 4 weeks, respectively. The film was clinically evaluated in patients with oral pain. IM at concentrations of 0.5% and 1% provided optimum analgesic effects, and the effects were the greatest in the 1% IM group. The addition of PEG to the nonadhesive layer reduced the number of patients experiencing discomfort at the site where the film was applied. Therefore this film formulation may be useful for local analgesic application due to its low dose requirement, moderate adhesion, and comfortable texture.


Increased expression of Cyclin F in liver cancer predicts poor prognosis: A study based on TCGA database.
Cyclin F (CCNF) dysfunction has been implicated in various forms of cancer, offering a new avenue for understanding the pathogenic mechanisms underlying hepatocellular carcinoma (HCC). We aimed to evaluate the role of CCNF in HCC using publicly available data from The Cancer Genome Atlas (TCGA). We used TCGA data and Gene Expression Omnibus (GEO) data to analyze the differential expression of CCNF between tumor and adjacent tissues and the relationship between CCNF and clinical characteristics. We compared prognosis of patients with HCC with high and low CCNF expression and constructed receiver operating characteristic (ROC) curves. In addition, we also explored the types of gene mutations in relevant groups and conducted Gene Set Enrichment Analysis (GSEA). The expression of CCNF in liver cancer tissues was significantly increased compared with that in adjacent tissues, and patients with high CCNF expression had a worse prognosis than those with low CCNF expression. Patients with high CCNF expression also had more somatic mutations. High expression of CCNF hampers the prognosis independently. The GSEA showed that the "http://www.gsea-msigdb.org/gsea/msigdb/cards/BIOCARTA_WNT_PATHWAY" Wnt pathway, "http://www.gsea-msigdb.org/gsea/msigdb/cards/BIOCARTA_P53_PATHWAY" P53 pathway, "http://www.gsea-msigdb.org/gsea/msigdb/cards/HALLMARK_PI3K_AKT_MTOR_SIGNALING" PI3K/Akt/mTOR pathway, "http://www.gsea-msigdb.org/gsea/msigdb/cards/HALLMARK_NOTCH_SIGNALING" Notch pathway were enriched in patients with the high CCNF expression phenotype. High CCNF expression can be seen as an independent risk factor for poor survival in HCC. Its expression may serve as a target for the diagnosis and treatment of liver cancer.


Treatment of lymphocyte depletion Hodgkin's disease in children.
Lymphocyte depletion Hodgkin's disease (LDHD) was diagnosed in 8 of 40 children with Hodgkin's disease treated from July 1969 to September 1976. All of the children received conventional radiotherapy and combination chemotherapy with mustine, vinblastine, procarbazine, and prednisone (MVPP). In LDHD children the disease was associated with severe anemia, splenomegaly presence of mediastinal enlargement, and dissemination at the time of diagnosis. Three children with LDHD did not achieve remission and died with uncontrolled disease. In two others, relapses occurred and led to death in one. Only three children remain in their first remission. Only 3 of 32 patients with non-LDHD relapsed and entered second remission. All 32 patients were free of disease at the time of evaluation, with mean survival of 88.2 months. We conclude that LDHD in childhood is frequently associated with clinically advanced disease and still has a bad prognosis. Alternative approaches to its therapy are needed.


Zinc oxide nanoparticle energy band gap reduction triggers the oxidative stress resulting into autophagy-mediated apoptotic cell death.
The physico-chemical properties of nanoparticle (NP), such as particle size, surface defects, crystallinity and accessible surface, affect NP photocatalytic activity that in turn defines the NP cytotoxic propensity. Since zinc oxide nanoparticle (ZnONP) energy band gap falls in a range of a semiconductor, the particle possesses photocatalytic activity. Hence, the study correlates energy band gap with cytotoxic propensity of ZnONP. To this end, ZnONPs with varying energy band gap are fabricated by varying calcination temperature. Cytotoxic propensity of the fabricated ZnONPs against HT1080 cell indicates that the particle with least energy band gap shows highest cytotoxicity. The data also indicate that the cytotoxicity is triggered primarily through reactive oxygen species (ROS)-mediated pathway. Additionally, the comet assay and γH2AX activity assay reveal that decreasing energy band gap of the particle increases DNA damaging propensity. Furthermore, cell cycle analysis indicates that the cell treatment with decreasing energy band gap ZnONP results in significant increase in cell population fraction in subG1 phase. Whereas, acridine orange binding assay and increased expression level of LC3II indicate that the cell tries to recover the stress by scavenging damaged cellular biomolecules and ROS using autophagosomes. Nevertheless, cell with the non-recoverable damages led into apoptotic cell death, as confirmed by Annexin V apoptosis assay, DNA fragmentation assay and 4,6-Diamidino-2-phenylindole dihydrochloride (DAPI) staining.


Site-Specific Introduction of Bioorthogonal Handles to Nanopores by Genetic Code Expansion.
Site-specific functionalization of natural amino acid-containing biological nanopores is pivotal in single molecule sensing. However, pore engineering methodologies are restricted to a limited choice and introduction of unnatural chemical components is extremely difficult. Herein we report the genetic code expansion (GCE) strategy to introduce unnatural amino acid (UAA) to an octameric Mycobacterium smegmatis porin A (MspA) nanopore. GCE allows for rapid and efficient introduction of bioorthogonal reactive site (i.e., azide) to the pore rim, and conjugation of single stranded DNA or lysozyme was demonstrated. The lysozyme-conjugated pore was further used for the discrimination of different oligosaccharides, demonstrating a sensing capacity that a bare MspA nanopore does not possess. GCE with bioorthogonal handles, which has never been previously applied in the preparation of nanopores, is a versatile strategy for pore engineering and may further expand the application scenarios of nanopores.


Zn-limited diet modifies the expression of the rate-regulatory enzymes involved in phosphatidylcholine and cholesterol synthesis.
Suboptimal intake of Zn is one of the most common nutritional worldwide problems. Previously, we showed that Zn deficiency produces alterations in lung lipid metabolism in rats. We studied the effect of a Zn-limited (ZL) diet on the expression of the enzymes involved in phosphatidylcholine and cholesterol synthesis. After 2 months of treatment with a ZL diet we found important variations in the lipid content of Wistar male rats: triacylglycerol (TG) decreased 60% (P<0.001) while esterified cholesterol (EC), free cholesterol and phospholipids (PL) increased 66%, 24 % and 25% respectively. We also observed a decrease of 40 % in the amount of (3)H incorporated into TG and an increase of 47% and 28% in the (3)H incorporated to PL and EC respectively. Fatty acid synthase and glucose-6-phosphate dehydrogenase activity was increased (P<0.01 and P<0.05 respectively). Glycerol-3-phosphate acyltransferase, lipoprotein lipase, diacyl glycerol acyl transferase and 3-hydroxy-3-methylglutaryl CoA reductase expression decreased (P<0.01 in all cases), while acetyl CoA carboxylase and cholinephosphate cytidylyltransferase increased (P<0.01 and P<0.005 respectively). These results suggest that ZL alters the expression of enzymes involved in phosphatidylcholine and cholesterol synthesis, which could lead to increased PL and cholesterol and decreased TG. This study suggests that major changes in the lipid composition of lung are induced by a ZL condition. Therefore, Zn deficiency must be taken into account in order to design therapies and public health interventions, such as Zn supplementation for high-risk subjects or certain diseases, such as asthma.


Constitutional mislocalization of Pten drives precocious maturation in oligodendrocytes and aberrant myelination in model of autism spectrum disorder.
There is a strong genetic association between germline PTEN mutation and autism spectrum disorder (ASD), making Pten-mutant models exemplary for the study of ASD pathophysiology. We developed the Ptenm3m4 mouse, where Pten is largely restricted from the nucleus, which recapitulates patient-like, autism-related phenotypes: behavioral changes, macrocephaly, and white matter abnormalities. This study aimed to investigate the contribution of oligodendrocyte (OL) lineage differentiation and functional changes in myelination to the white matter phenotype. OL lineage differentiation and myelination in Ptenm3m4 mice was studied using immunohistochemical and electron microscopic analyses. We also used primary oligodendrocyte progenitor cells (OPCs) to determine the effect of the Ptenm3m4 mutation on OPC proliferation, migration and maturation. Finally, we assessed the myelinating competency of mutant OLs via co-culture with wildtype dorsal root ganglia (DRG) neurons. The in vivo analyses of Ptenm3m4/m3m4 murine brains showed deficits in proteolipid protein (Plp) trafficking in myelinating OLs. Despite the increased expression of myelin proteins in the brain, myelin deposition was observed to be abnormal, often occurring adjacent to, rather than around axons. Mutant primary OPCs showed enhanced proliferation and migration. Furthermore, mutant OPCs matured precociously, exhibiting aberrant myelination in vitro. Mutant OPCs, when co-cultured with wildtype DRG neurons, showed an inability to properly ensheath axons. Our findings provide evidence that the Ptenm3m4 mutation disrupts the differentiation and myelination programs of developing OLs. OL dysfunction in the Ptenm3m4 model explains the leukodystrophy phenotype, a feature commonly associated with autism, and highlights the growing importance of glial dysfunction in autism pathogenesis.


Plasma fetuin-a levels and the risk of myocardial infarction and ischemic stroke.
Fetuin-A, a protein almost exclusively secreted by the liver, induces insulin resistance and subclinical inflammation in rodents. Circulating fetuin-A levels are elevated in humans with metabolic syndrome and insulin resistance, conditions that are associated with increased risk of cardiovascular disease. We investigated the association between fetuin-A levels and the risk of future myocardial infarction (MI) and ischemic stroke (IS) in a case-cohort study based on the European Prospective Investigation into Cancer and Nutrition (EPIC)-Potsdam Study comprising 27 548 middle-aged subjects of the general population. Fetuin-A levels were measured in plasma of 227 individuals who developed MI, in 168 who developed IS, and in 2198 individuals who remained free of cardiovascular events during a mean follow-up of 8.2+/-2.2 years. Individuals in the highest compared with the lowest quintile of plasma fetuin-A had significantly increased risks of MI (relative risk, 3.80; 95% confidence interval, 2.37 to 6.10; P for trend <0.0001) and IS (relative risk, 3.93; 95% confidence interval, 2.17 to 7.12; P for trend <0.0001) after adjustment for sex and age. Additional adjustment for smoking status, body mass index, waist circumference, alcohol consumption, educational attainment, physical activity, hypertension, diabetes mellitus, total and high-density lipoprotein cholesterol, and C-reactive protein only moderately attenuated these risks (MI: relative risk, 3.25; 95% confidence interval, 2.01 to 5.28; P for trend <0.0001; IS: relative risk, 3.78; 95% confidence interval, 2.06 to 6.94; P for trend <0.0001). Our data provide evidence for a link between high plasma fetuin-A levels and an increased risk of MI and IS. Therefore, more research is warranted to determine the role of fetuin-A in the pathophysiology of cardiovascular disease.


Heterogeneous distribution and replication activity of mitochondria in Xenopus laevis oocytes.
In early diplotene oocytes of Xenopus laevis mitochondria are not dispersed all over the cytoplasm but gathered in a well described mitochondrial mass [18]. When tracing these organelles during active vitellogenesis we observe that some of them are involved in the elaboration of a cortical layer at the vegetative hemisphere of the cell while others stay around the nucleus. The latter contribute to the transient formation of a mitochondrial crown throughout active mitochondriogenesis. Autoradiographic studies of thymidine incorporation into mtDNA suggest a differential participation of each organelle to the final population of a full-grown oocyte according to its position in the cytoplasm.


Safety risks in the ambulatory setting.
Over the past decade, the focus of safety implementation has been on hospital settings, and most research on patient safety has examined hospital care. Yet, each year, 300 Americans are seen in ambulatory settings for every 1 person admitted to a hospital, and research shows that errors in ambulatory settings can be just as devastating as those in hospitals, and, as in the hospital setting, ambulatory errors or events often trigger legal action. The American Medical Association's report summarizing and compiling the past decade's research identifies 3 general gaps in the current research that impede safety analysis and 6 errors that are most common in ambulatory care that warrant attention. As new models of care emerge with an increased focus on continuity across care settings, there are also nascent opportunities for risk managers to analyze and evaluate ambulatory safety, implement strategies, and develop and test tools that could result in safer patient outcomes.


Grieving experiences amongst adolescents orphaned by AIDS: Analysis from event history calendars.
Mental health is an essential component of adolescent health and wellbeing. Mental health practitioners assess adolescents' mental health status to identify possible issues that may lead to mental health problems. However, very few of the tools used to assess the mental health status of adolescents include assessment for grieving and coping patterns. The current tools used for assessing an individual's mental health are lengthy and not comprehensive. The purpose of this study was to assess grieving patterns of adolescents orphaned by AIDS and to appraise the usefulness of an event history calendar as an assessment tool for identifying grieving experiences, in order to guide and support these adolescents through the grieving process. One hundred and two adolescents aged 14-18 years, who had been orphaned by AIDS, completed an event history calendar, reviewed it with the researcher and reported their perceptions of it. Thematic analysis of the event history calendar content revealed that it is an effective, time-efficient, adolescent-friendly tool that facilitated identification and discussion of the orphaned adolescents' grieving patterns. Crying, isolation, silence and violent outbursts were the main grieving patterns reported by adolescents orphaned by AIDS. The researcher recommends use of the event history calendar for identification of orphaned adolescents' grieving experiences. Early identification would enable mental health practitioners to support them in order to prevent the occurrence of mental illness due to maladaptive grieving.


Adrenaline inhibits adenosine diphosphate induced intravascular aggregation of rat platelets through stimulation of alpha 2 adrenoceptors.
The aim was to determine the receptors involved in the antiaggregatory effects of adrenaline in the rat. The 111indium oxine technique was used to evaluate ADP induced pulmonary accumulation of 111In labelled platelets in adrenalectomised rats (ie, no endogenous adrenaline) in the absence of anticoagulants. Adrenaline was infused (10, 20 and 40 micrograms.kg-1.h-1) and ADP aggregation measured. Adrenoceptor agonists, isoprenaline, methoxamine, and BHT 933, were given to evaluate whether they mimic the adrenaline effects. Adrenoceptor antagonists, yohimbine, propranolol, prazosin, and WY 26392, were given to assess whether they block the adrenaline effects. In all cases, pulmonary accumulation of platelets was measured in response to ADP. Adrenaline dose dependently inhibited platelet aggregation. BHT 933 (1.5 micrograms.kg-1.min-1) mimicked, while phentolamine (1 mg.kg-1), yohimbine (2.5 mg.kg-1), and WY 26392 (0.1, 1.0, 5.0 mg.kg-1) blocked, the adrenaline effects. Methoxamine (3.3 micrograms.kg-1.min-1) produced a relatively weak non-specific inhibition of ADP aggregation. Isoprenaline (33 ng.kg-1.min-1) did not mimic the effects of adrenaline, neither did propranolol (1 mg.kg-1) affect the adrenaline inhibition of the aggregatory responses to ADP. Indomethacin (3 mg.kg-1) blocked the inhibitory effects of adrenaline. Adrenaline stimulates alpha receptors on platelets but inhibits platelet aggregation. There is no evidence to suggest that beta receptors participated in the effects produced by adrenaline. There is a role for cyclo-oxygenase products in the inhibitory effects of adrenaline on aggregation.


Anisotropic Finite Element Modeling Based on a Harmonic Field for Patient-Specific Sclera.
Purpose. This study examined the influence of anisotropic material for human sclera. Method. First, the individual geometry of patient-specific sclera was reproduced from a laser scan. Then, high quality finite element modeling of individual sclera was performed using a convenient automatic hexahedral mesh generator based on harmonic field and integrated with anisotropic material assignment function. Finally, comparison experiments were designed to investigate the effects of anisotropy on finite element modeling of sclera biomechanics. Results. The experimental results show that the presented approach can generate high quality anisotropic hexahedral mesh for patient-specific sclera. Conclusion. The anisotropy shows significant differences for stresses and strain distribution and careful consideration should be given to its use in biomechanical FE studies.


Exploring gender and identity issues among female adolescent and young adults who connect in an anonymous platform.
There has been an increased usage of online cancer support groups as a resource for health-related information and social support. This work analyzes message blog posts from an anonymous online support community to better understand issues related to gender and identity among female adolescent and young adults. This work sheds further light into the nuances of gender and identity issues including motherhood and reproductive issues, physical appearance, and romantic relationships. Specifically, findings reveal that female adolescent and young adults experience issues pertaining to infertility, feeling like a bad mom, hair loss, scarring, dating, and intimacy. These findings of this work offer further guidance about how healthcare providers and caregivers can attempt to meet the needs of female adolescent and young adults.


Mucoepidermoid carcinoma in the mandible: report of case.
Mucoepidermoid carcinoma, a salivary gland tumor arising centrally within the jaws, is rare. The literature concerning this tumor has been reviewed. Diagnosis and treatment of a case in 44-year-old man has been presented.


Imaging abusive head trauma: why use both computed tomography and magnetic resonance imaging?
Abusive head trauma is the leading cause of death in child abuse cases. The majority of victims are infants younger than 1 year old, with the average age between 3 and 8 months, although these injuries can be seen in children up to 5 years old. Many victims have a history of previous abuse and the diagnosis is frequently delayed. Neuroimaging is often crucial for establishing the diagnosis of abusive head trauma as it detects occult injury in 37% of cases. Several imaging patterns are considered to be particularly associated with abusive head trauma. The presence of subdural hematoma, especially in multiple locations, such as the interhemispheric region, over the convexity and in the posterior fossa, is significantly associated with abusive head trauma. Although CT is the recommended first-line imaging modality for suspected abusive head trauma, early MRI is increasingly used alongside CT because it provides a better estimation of shear injuries, hypoxic-ischemic insult and the timing of lesions. This article presents a review of the use and clinical indications of the most pertinent neuroimaging modalities for the diagnosis of abusive head trauma, emphasizing the newer and more sensitive techniques that may be useful to better characterize the nature and evolution of the injury.


Antibiotic Consumption Trends in China: Evidence From Six-Year Surveillance Sales Records in Shandong Province.
The overuse of antibiotics is a serious public health problem in China, causing a high rate of antimicrobial resistance. This study identified the trends of antibiotic consumption in China to provide evidence for further intervention. The six-year surveillance data on antibiotic sales from 2012 to 2017, which served as a proxy for consumption, were collected from 39 public health care facilities in Shandong province, including three tertiary hospitals, six secondary hospitals, and 30 primary health centers. Based on the Anatomical Therapeutic Chemical (ATC)/DDD methodology, antibiotic consumption was formulated in defined daily doses (DDD) per 1,000 inhabitants per day (DID). The total antibiotic consumption among all health care settings increased from 16.07 DID in 2012 to a peak of 17.44 DID in 2015 and then decreased to 11.35 DID in 2017 with a 34.90% reduction. J01C (beta-lactam antimicrobials, penicillin), the most frequently used antibiotic class, accounted for 36.32% of the total DID. Consumption of carbapenems increased from 0.029 DID in 2012 to 0.08 DID in 2017. Parenteral antibiotics accounted for nearly 40% of the total consumption. Compared with the 2012 figures, the 2017 consumption showed a small increase in hospital sector that was compensated by the decrease in community care. A substantial reduction in total antibiotic consumption was observed in China from 2012 to 2017. However, the extensive consumption of broad-spectrum antimicrobials, high proportion of parenteral antibiotic use, and increased use of last-resort antibiotics attracted public health concerns.


Feed-forward neural network as nonlinear dynamics integrator for supercontinuum generation: erratum.
We present an erratum to our Letter [Opt. Lett.47, 802 (2022)10.1364/OL.448571]. This erratum corrects an error in the sign of one of the higher-order dispersion coefficient used in the simulations of Figs. 2 and 4, as well as in Figs. S1 and S3. The simulations in the original Letter were performed using the correct value, and therefore this correction does not affect any of the results and conclusions of the original Letter.


Ethical challenges in social media engagement and research: considerations for code of engagement practices.
Social media have great potential for effectively communicating about public health risks so people make healthier and safer choices. If used appropriately, social media can strengthen trust between the public and the institution. If used inappropriately, it may also create distrust. This note addresses some of the ethical challenges in using social media for communication and research. It reflects on opportunities in social media risk communication based on experience from the World Health Organization (WHO) and suggests a code of engagement be included in corporate social media policies that contain guidance as to what conduct is or is not appropriate with a view to maintaining public trust in the institution. The note concludes with considerations about the ethical use of social media in research, which is particularly relevant for entities communicating about ionizing radiation, including during emergency situations.


Lithium-ion electrolytic substrates for sub-1V high-performance transition metal dichalcogenide transistors and amplifiers.
Electrostatic gating of two-dimensional (2D) materials with ionic liquids (ILs), leading to the accumulation of high surface charge carrier densities, has been often exploited in 2D devices. However, the intrinsic liquid nature of ILs, their sensitivity to humidity, and the stress induced in frozen liquids inhibit ILs from constituting an ideal platform for electrostatic gating. Here we report a lithium-ion solid electrolyte substrate, demonstrating its application in high-performance back-gated n-type MoS2 and p-type WSe2 transistors with sub-threshold values approaching the ideal limit of 60 mV/dec and complementary inverter amplifier gain of 34, the highest among comparable amplifiers. Remarkably, these outstanding values were obtained under 1 V power supply. Microscopic studies of the transistor channel using microwave impedance microscopy reveal a homogeneous channel formation, indicative of a smooth interface between the TMD and underlying electrolytic substrate. These results establish lithium-ion substrates as a promising alternative to ILs for advanced thin-film devices.


Neurological channelopathies: new insights into disease mechanisms and ion channel function.
Inherited mutations of ion channels provide unique insights into the mechanisms of many neurological diseases. However, they also provide a wealth of new information on the fundamental biology of ion channels and on neuron and muscle function. Ion channel genes are continuing to be discovered by positional cloning of disease loci. And some mutations provide unique tools to manipulate signalling cascades, which cannot be achieved by pharmacological intervention. Here we highlight some unanswered questions, and some promising areas for research that will likely lead to a fuller understanding of the link from molecular lesion to disease.


LXR activation by GW3965 alters fat tissue distribution and adipose tissue inflammation in ob/ob female mice.
To investigate the role of liver X receptor (LXR) in adipose tissue metabolism during obesity, ob/ob mice were treated for 5 weeks with the synthetic LXR agonist GW3965. MRI analysis revealed that pharmacological activation of LXR modified fat distribution by decreasing visceral (VS) fat and inversely increasing subcutaneous (SC) fat storage without affecting whole body fat content. This was concordant with opposite regulation by GW3965 of the lipolytic markers hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) in the two fat depots; moreover, the expression of genes involved in lipogenesis was significantly induced in SC fat. Lipidomic analysis suggested that changes in lipid composition in response to GW3965 also varied between VS and SC fat. In both depots, the observed alteration in lipid composition indicated an overall change toward less lipotoxic lipids. Flow cytometry analysis showed decreased immune cell infiltration in adipose tissue of ob/ob mice in response to GW3965 treatment, which in VS fat mainly affected the macrophage population and in SC fat the lymphocyte population. In line with this, the expression and secretion of proinflammatory markers was decreased in both fat deposits with GW3965 treatment.


[Advances on network pharmacology in ethnomedicine research].
Ethnomedicine is the precious wealth left by ethnic minorities in their struggle against diseases. It is similar to traditional Chinese medicine in a narrow sense and has the characteristics of multi-component,multi-target and multi-channel synergy. Under the guidance of the theory of ethnomedicine,the combination of ethnomedicine and network pharmacology will help to understand the essence of the prevention and treatment of ethnomedicines in a dynamic and holistic manner. This paper reviews the research progress of network pharmacology applied in ethnomedicine,analyses the problems and challenges existing in the application of network pharmacology in ethnomedicine research at present,such as inaccurate data and information,lack of network analysis platform for effective analysis of dose-effect relationship of chemical constituents and weak basic research of ethnomedicine,and puts forward corresponding prospects.


The knockdown of H19lncRNA reveals its regulatory role in pluripotency and tumorigenesis of human embryonic carcinoma cells.
The function of imprinted H19 long non-coding RNA is still controversial. It is highly expressed in early embryogenesis and decreases after birth and re-expressed in cancer. To study the role of H19 in oncogenesis and pluripotency, we down-regulated H19 expression in vitro and in vivo in pluripotent human embryonic carcinoma (hEC) and embryonic stem (hES) cells. H19 knockdown resulted in a decrease in the expression of the pluripotency markers Oct4, Nanog, TRA-1-60 and TRA-1-81, and in the up-regulation of SSEA1; it further attenuated cell proliferation, decreased cell-matrix attachment, and up-regulated E-Cadherin expression. SCID-Beige mice transplanted with H19 down-regulated hEC cells exhibited slower kinetics of tumor formation, resulting in an increased animal survival. Tumors derived from H19 down-regulated cells showed a decrease in the expression of pluripotency markers and up-regulation of SSEA-1 and E-cadherin. Our results suggest that H19 oncogenicity in hEC cells is mediated through the regulation of the pluripotency state.


Negative electrodes for supercapacitors with good performance using conductive bismuth-catecholate metal-organic frameworks.
Metal-organic frameworks (MOFs) have attracted increasing research interest in various fields. Unfortunately, the poor conductivity of most traditional MOFs considerably hinders their application in energy storage. Benefiting from the full charge delocalization in the atomic plane, two-dimensional conductive coordination frameworks achieve good electrochemical performance. In this work, π-π coupling conductive bismuth-catecholate nanobelts with tunable lengths, Bi(HHTP) (HHTP = 2,3,6,7,10,11-hexahydroxytriphenylene), are synthesized by a simple hydrothermal reaction and their length-dependent electrochemical properties are also investigated. The Bi(HHTP) nanobelts (about 10 μm in length) possess appropriate porosity, numerous redox active sites and good electrical conductivity. Being a negative electrode for supercapacitors, Bi(HHTP) nanobelts display a high specific capacitance of 234.0 F g-1 and good cycling stability of 72% after 1000 cycles. Furthermore, the mechanism of charge storage is interpreted for both battery-type and surface-capacitive behavior. It is believed that the results of this work will help to develop battery-type negative electrode materials with promising electrochemical performance using some newly designed π-π coupling conductive coordination frameworks.


[Computer-assisted measurement of intracranial pressure. The Pic Part system].
In this article the authors describe a computer assisted pressure measuring equipment: the PIC PART system which allows real time visualisation of intracranial and possibly arterial pressure, and above all the automatic analysis of measures after long casting recording (24 hours).


Maintenance of genetic variability in a specific pathogen-free breeding colony.
We used 18 genetic loci including blood groups, isozymes, and a serum protein to evaluate our efforts to preserve genetic variability in a specific pathogen-free (SPF) colony of rhesus monkeys. We compared genetic variability in the SPF population to the virally contaminated, non-SPF population from which it was derived. There was no change in the average gene diversity between the SPF and non-SPF populations. However, gene diversity at blood group Q locus increased significantly in the SPF population, while blood group M locus showed an insignificant trend toward decreased gene diversity. Allele frequencies changed significantly at blood group Q locus, although no alleles were lost from the population. We hypothesized that this change was due to extensive overreproduction by a small number of founder males that possessed the initially rare allele, Q1. There was no evidence that this change was associated with genes involved in viral infection.


Surgical approaches to acoustic nerve tumours.
A critical account is given of the orthodox neurosurgical approach to acoustic nerve tumours and alternatives through the petrous bone or tentorium.


Discrimination between normal and cancer white blood cells using holographic projection technique.
White blood cells (WBCs) play a vital role in the diagnosis of many blood diseases. Such diagnosis is based on the morphological analysis of blood microscopic images which is performed manually by skilled hematologist. However, this method has many drawbacks, such as the dependence on the hematologist's skill, slow performance, and varying accuracy. Therefore, in the current study, a new optical method for discrimination between normal and cancer WBCs of peripheral blood film (PBF) images is presented. This method is based on holographic projection technique which is able to provide an accurate and fast optical reconstruction method of WBCs floating in the air. Besides, it can provide a 3D visualization map of one WBC with its characterization parameters from only a single 2D hologram. To achieve that, at first, WBCs are accurately segmented from the microscopic PBF images using a developed in-house MATLAB code. Then, their associated phase computer-generated holograms (CGHs) are calculated using the well-known iterative Fourier transform algorithm (IFTA). Within the utilized algorithm, a speckle noise reduction technique, based on temporal multiplexing of spatial frequencies, is applied to minimize the speckle noise across the reconstruction plane. Additionally, a special hologram modulation is added to the calculated holograms to provide a 3D visualization map of one WBC, and discriminate normal and cancer WBCs. Finally, the calculated phase-holograms are uploaded on a phase-only spatial light modulator (SLM) for optical reconstruction. The optical reconstruction of such phase-holograms yields precise representation of normal and cancer WBCs. Moreover, a 3D visualization map of one WBC with its characterization parameters is provided. Therefore, the proposed technique can be used as a valuable tool for interpretation and analysis of WBCs, this in turn could provide an improvement in diagnosis and prognosis of blood diseases.


Time course of morphological alterations of fungiform papillae and taste buds following chorda tympani transection in neonatal rats.
The time course of structural changes in fungiform papillae was analyzed in rats that received unilateral chorda tympani nerve transection at 10 days of age. Morphological differences between intact and denervated sides of the tongue were first observed at 8 days postsection, with an increase in the number of fungiform papillae that did not have a pore. In addition, the first papilla with a filiform-like appearance was noted on the denervated side at 8 days postsectioning. By 11 days after surgery, the total number of papillae and the number of papillae with a pore were significantly lower on the transected side of the tongue as compared to the intact side. At 50 days postsection, there was an average of 70.5 fungiform papillae on the intact side and a mean of only 20.8 fungiform papillae the denervated side. Of those few remaining papillae on the cut side, an average of 13.5 papillae were categorized as filiform-like, while no filiform-like papillae occurred on the intact side. Significant reduction in taste bud volume was noted at 4 days posttransection and further decrements in taste bud volume were noted at 8 and 30 days postsection. Electron microscopy of the lingual branch of the trigeminal nerve from adult rats that received neonatal chorda tympani transection showed normal numbers of both myelinated and unmyelinated fibers. Thus, in addition to the well-characterized dependence of taste bud maintenance on the chorda tympani nerve, the present study shows an additional role of the chorda tympani nerve in papilla maintenance during early postnatal development.


Double Soft-Template Synthesis of Nitrogen/Sulfur-Codoped Hierarchically Porous Carbon Materials Derived from Protic Ionic Liquid for Supercapacitor.
Heteroatom-doped hierarchical porous carbon materials derived from the potential precursors and prepared by a facile, effective, and low-pollution strategy have recently been particularly concerned in different research fields. In this study, the interconnected nitrogen/sulfur-codoped hierarchically porous carbon materials have been successfully obtained via one-step carbonization of the self-assembly of [Phne][HSO4] (a protic ionic liquid originated from dilute sulfuric acid and phenothiazine by a straightforward acid-base neutralization) and the double soft-template of OP-10 and F-127. During carbonization process, OP-10 as macroporous template and F-127 as mesoporous template were removed, while [Phne][HSO4] not only could be used as carbon, nitrogen, and sulfur source, but also as a pore forming agent to create micropores. The acquired carbon materials for supercapacitor not only hold a large specific capacitance of 302 F g-1 even at 1.0 A g-1, but also fine rate property with 169 F g-1 at 10 A g-1 and excellent capacitance retention of nearly 100% over 5000 circulations in 6 M KOH electrolyte. Furthermore, carbon materials also present eximious rate performance with 70% in 1 M Na2SO4 electrolyte.


Doses of solar ultraviolet radiation correlate with skin cancer rates in Japan.
We analyzed trends in the disease rate of skin cancers in the 1976-80 and 1986-90 intervals in the 27 university hospitals in Japan. We also measured doses of solar ultraviolet (UV) radiation at Sapporo, Kobe and Miyazaki to evaluate the relationship between the two in Japan. The rates of basal cell carcinoma (BCC) and actinic keratosis (AK) were higher in 1986-90 than in 1976-80, whereas the rate of squamous cell carcinoma (SCC) was lower in 1986-90 than in the earlier period. The rates of SCC, BCC and AK in the southern part of Japan were about five times higher than those in the north, and the average daily UV dose measured with a Robertson-Berger meter in 1995 was about 1.8 times higher in Miyazaki than in Kobe. That measured by MS-210D UV dosimeter in Sapporo was about 0.53 times lower than in Kobe. These results demonstrate that solar UV dose is higher in the southern part of Japan than that in the northern part, explaining the higher rate of non-melanoma skin cancer in southern part of Japan. A significant increase of AK and BCC may reflect the trend of UV increase in Japan.


2-Hydroxybenzohydrazide as a novel potential candidate against nociception, inflammation, and pyrexia: in vitro, in vivo, and computational approaches.
The current study was designed to evaluate the 2-hydroxybenzohydrazide (HBH) as a drug having efficacy against pyrexia, inflammation, and nociception. Besides, the therapeutic effects of HBH on oxidative stress and C-reactive proteins were also evaluated. The pharmacological studies on HBH (20-60 mg/kg) were conducted using nociception, inflammation, and pyrexia standard models. Naloxone antagonism was performed to assess the possible involvement of opioidergic mechanisms. The antioxidant study was conducted on ABTS and DPPH assays using gallic acid as a standard. Moreover, the binding capability of HBH with enzymes cyclooxygenase-I/II (COX-I/II) was determined using molecular modeling analysis. The findings indicated that the HBH dose-dependently inhibited pain, inflammation, and pyrexia. The HBH has significant anti-nociceptive and anti-inflammatory activities at 60 mg/kg (***p < 0.001), similar to the lower doses of diclofenac sodium (50 mg/kg) and tramadol (30 mg/kg). The HBH at 60 mg/kg reduced pyrexia as paracetamol (150 mg/kg). The HBH at 20-60 mg/kg doses declined the plasma C-reactive protein concentration. The mechanistic studies showed that the anti-nociceptive effect of HBH was antagonized by naloxone, indicating that the opioidergic mechanisms are involved. Furthermore, computational studies showed that the HBH exhibited an affinity for COX-I/II target receptors. The HBH significantly inhibited ABTS and DPPH radicals (IC50 = 33.81 and 26.74 μg/ml). These results proposed that the HBH has significant antipyretic, anti-inflammatory, and anti-nociceptive activities involving opioidergic mechanism.


The relationship of brachiocephalic vessel atheroma to transient ischemia of the brain and retina.
The relationships between transient cerebral and retinal ischemic symptoms and brachiocephalic angiographic findings were studied in 464 patients. The arterial abnormalities were more often bilateral than the symptoms suggested. Normal angiographic appearances were common, although abnormal arteries in patients without related symptoms were also common. When stenosis of the carotid system was 75 per cent or more, there was a strong correlation with symptoms.


Indications for and results of nephrectomy for metastatic renal cell carcinoma.
We find that, when metastases are present, a nephrectomy for renal cell carcinoma is not justified in the vast majority of cases. The only noncontroversial indication is for palliative nephrectomy which is performed to relieve intractable symptoms in the properly chosen patient. However, it is unusual to see a patient with tumor-related symptoms that cannot be conservatively managed. A relative indication for nephrectomy is found in the patient with osseous metastases only, as some studies have demonstrated a prolongation of survival by adjunctive nephrectomy in this setting. Another relative indication is in the patient with limited metastases that are amenable to surgical or radiation ablation coupled with nephrectomy; patients in whom the metastasis appears years after the nephrectomy have the best chance for a successful outcome. When therapy that is effective against metastatic tumor is eventually found, adjunctive nephrectomy as a debulking procedure may become indicated; until then, surgery is not justified. On the basis of data presently available, adjunctive nephrectomy in the hope of inducing spontaneous regression of metastases is never indicated.


Bone effects of hexarelin, a GH-releasing peptide, in female rats: influence of estrogen milieu.
The present study was performed to evaluate the potential influence of the estrogen milieu in modulating the effects of GH/IGF stimulation by a GH-releasing peptide, hexarelin (HEXA), on bone metabolism and mineral density in middle-aged female rats. HEXA was administered for 60 days (50 microg/kg s.c. twice a day) to intact and ovariectomized (OVX) 11-month-old female rats and changes in bone parameters were evaluated with respect to those of the same rats under baseline conditions and with those of control rats (intact and OVX) administered isovolumetric amounts of physiological saline. Serum total alkaline phosphatase (ALP) and urinary deoxypyridinoline (Dpd) were measured before and at various times during HEXA treatment. Bone mineral content (BMC) and density of lumbar vertebrae and femoral mid-diaphyses were measured by dual energy X-ray absorptiometry before and after treatment. In all groups, serum IGF-I levels were determined before and during treatment and the GH secretory response to HEXA was assessed at the end of the experiment. In intact rats, HEXA did not modify Dpd urinary excretion, induced a trend toward an increase of serum ALP activity and significantly increased BMC (+6.5%) and bone area (+4.1%) only at lumbar vertebrae. In OVX rats, HEXA did not modify the OVX-induced increase in bone turnover markers (Dpd and ALP) and did not affect the OVX-induced vertebral bone loss, but significantly increased BMC (+7.2%) and bone area (+5.3%) at femoral mid-diaphyses. HEXA significantly increased serum IGF-I levels at day 14, but not at day 60, in both intact and OVX rats, whereas the GH secretory response to HEXA was higher in the former than in the latter. Overall, the present data demonstrate that chronic HEXA treatment increases BMC and bone area at lumbar vertebrae in intact rats and at femoral diaphyses in OVX rats. The different sensitivity to HEXA of the skeletal districts examined is related to the estrogen milieu and may reflect a complex interplay between estrogens and GH/IGF function.


Efficient algorithms for detecting changes in intraventricular electrogram morphology.
Computing a correlation coefficient between a stored template of a normal beat and subsequent beats is a widely used indicator of changes in electrogram morphology. While effective, this method is quite computationally demanding, particularly for use in implantable devices with limited available power. We investigated methods for computing a normalized autocorrelation as efficient template matching alternatives to the correlation coefficient. These four algorithms, polarity incidence (PI), hybrid sign (HS), modified hybrid sign (MHS), and relative magnitude (RM) are compared with the correlation coefficient (CC) for detecting changes in intraventricular electrogram morphology. We sought to determine the number of instances in which 75% (or 90%) of a patient's sinus rhythm beats could be distinguished from 75% (or 90%) of the patient's ventricular tachycardia beats at the 95% confidence level in 30 patients. The relative magnitude method performs nearly as well as the correlation coefficient in nearly every case but requires only 3 multiplies per application. Hence the relative magnitude algorithm may be more appropriate for implantable devices.


The emerging role of auxins as bacterial signal molecules: Potential biotechnological applications.
Microorganisms are exposed in their natural niches to a wide diversity of signal molecules. Specific detection of these signals results in alterations in microbial metabolism and physiology. Auxins like indole-3-acetic acid are key phytohormones that regulate plant growth and development. Nonetheless, auxin biosynthesis is not restricted to plants but is ubiquitous in all kingdoms of life. This wide phylogenetic distribution of auxins production, together with the diversity of regulated cellular processes, have made auxins key intra- and inter-kingdom signal molecules in life modulating, for example microbial physiology, metabolism and virulence. Despite their increasing importance as global signal molecules, the mechanisms by which auxins perform their regulatory functions in microorganisms are largely unknown. In this article, we outline recent research that has advanced our knowledge of the mechanisms of bacterial auxin perception. We also highlight the potential applications of this research in aspects such as antibiotic production, biosensor design, plant microbiome engineering and antivirulence therapies.


Pubertal gynecomastia.
Gynecomastia is a benign condition in males, characterized by proliferation of glandular elements resulting in concentric enlargement of one or both breasts. During puberty, there is often a transient relative imbalance between estrogen and testosterone, leading to gynecomastia. This condition usually resolves by age 18 years when adult androgen/estrogen ratios are achieved. Laboratory evaluation should include testosterone, estradiol, and gonadotropins; karyotype should be obtained in pubertal patients with testes volumes less than 6 ml. The mainstay in treatment of pubertal gynecomastia is still sympathetic reassurance considering the benign nature of the condition. Surgical removal of the breast glandular tissue should be considered in boys who have had persistent pubertal gynecomastia and have completed or nearly completed puberty.


Immunological studies of reactive oxygen species damaged catalase in patients with systemic lupus erythematosus: correlation with disease activity index.
This study was undertaken to investigate the status and contribution of oxidized catalase in systemic lupus erythematosus (SLE) and to explore whether oxidized catalase has a role in disease progression. Catalase (CAT) was modified by reactive oxygen species (ROS). Sera from 50 SLE patients with varying levels of disease activity according to SLE Disease-Activity-Index (SLEDAI) and 45 age- and sex-matched healthy controls were evaluated for antibodies against oxidized CAT. Serum analysis showed significantly higher level of anti-oxidized-CAT-antibodies in SLE patients compared with controls. Interestingly, not only was there an increased number of subjects positive for anti-oxidized-CAT-antibodies, but also the levels of these antibodies were significantly higher among SLE patients, whose SLEDAI scores were ≥ 10 as compared with lower SLEDAI scores (<10). In addition, significant correlation was observed between the levels of anti-oxidized-CAT-antibodies and SLEDAI score (r = 0.796). Furthermore, sera from SLE patients had lower levels of CAT activity compared with control sera. These findings support an association between oxidized CAT and SLE. The stronger response observed in serum samples from patients with higher SLEDAI scores suggests that oxidized CAT may be a useful biomarker in evaluating the progression of SLE and in elucidating the mechanisms of disease pathogenesis.


Natural treatment system models for wastewater management: a study from Hyderabad, India.
Wastewater generated on a global scale has become a significant source of water resources which necessitates appropriate management strategies. However, the complexities associated with wastewater are lack of economically viable treatment systems, especially in low- and middle-income countries. While many types of treatment systems are needed to serve the various local issues, we propose natural treatment systems (NTS) such as natural wetlands that are eco-friendly, cost-effective, and can be jointly driven by public bodies and communities. In order for it to be part of wastewater management, this study explores the NTS potential for removal of pollutants, cost-effectiveness, and reuse options for the 1.20 million m3/day of wastewater generated in Hyderabad, India. The pilot study includes hydro-geophysical characterization of natural wetland to determine pollutant removal efficiency and its effective utilization for treated wastewater in the peri-urban habitat. The results show the removal of organic content (76-78%), nutrients (77-97%), and microbes (99.5-99.9%) from the wetland-treated wastewater and its suitability for agriculture applications. Furthermore, the wetland efficiency integrated with engineered interventions led to the development of NTS models with different application scenarios: (i) constructed wetlands, (ii) minimized community wetlands, and (iii) single outlet system, suitable for urban, peri-urban and rural areas, respectively.


Does Age Influence the Risk of Incident Knee Osteoarthritis After a Traumatic Anterior Cruciate Ligament Injury?
The development of radiographic knee osteoarthritis (OA) after an anterior cruciate ligament (ACL) rupture has long been studied and proven in the adolescent population. However, similar exhaustive investigations have not been conducted in mature-aged athletes or in older populations. To identify whether an older adult population had an increased risk of incident radiographic knee OA after a traumatic knee injury compared with a young adult population. Cohort study; Level of evidence, 3. Patients with ACL ruptures who underwent primary reconstruction were enrolled in a prospective, longitudinal single-center study over 15 years. The adult cohort was defined as participants aged ≥35 years who had a knee injury resulting in an ACL tear, the adolescent-young cohort suffered similar knee injuries and were aged ≤25 years, and a third cohort of participants aged 26 to 34 years who suffered a knee injury was included to identify the existence of any age-related dose-response relationship for the onset of radiographic knee OA. A Kaplan-Meier survival analysis was employed to determine the occurrence of incident radiographic OA across the study populations at 2, 5, 10, and 15 years after reconstruction. Significance at each time point was analyzed using chi-square tests. A total of 215 patients, including 112 adolescents (mean age, 20.4 years; 50.9% female), 71 patients aged 26 to 34 years (mean age, 29.2 years; 42.3% female), and 32 adults (mean age, 40.2 years; 59.4% female), were assessed for International Knee Documentation Committee (IKDC) grading on knee radiographs. It was found that 53.0% and 77.8% of adults at a respective 10 and 15 years after reconstruction had an IKDC grade of B or greater compared with 17.7% and 61.6% of the adolescent-young cohort. Chi-square testing found that adults developed OA earlier than adolescents at 5 and 10 years after reconstruction (P = .017 and P < .0001, respectively). However, survival analysis did not demonstrate that adults were more likely to develop radiographic knee OA at 15 years after reconstruction compared with the adolescent-young cohort (P = .4). The age at which an ACL injury is sustained does not appear to influence the rate of incident radiographic knee OA, although mature-aged athletes are likely to arrive at the OA endpoint sooner.


A reply to McLachlan.
In an earlier article in this journal, I advanced five ethical arguments in favour of a voluntary, unpaid system of blood donation. In his reply to my article, Hugh McLachlan criticised one of those arguments, namely, the argument that an unpaid system promotes altruism and social solidarity. In this reply to Dr. McLachlan, I maintain that his criticism is misguided, and that he appears unclear not only about my own argument, but also about his own.


Signal transmission and covert prepattern in the metamorphosis of Hydractinia echinata (Hydrozoa).
Planulae are simply structured larvae lacking an overt longitudinal organization. In the course of a rapid metamorphosis, however, they transform into polyps, which display striking structural patterns. Metamorphosis takes place only in response to external stimuli. Surgical removal and transplantation of larval parts reveal that external stimuli, including artificial inducers such as cesium ions, tumor promoters and diacylglycerol, act on the anterior quarter of the larva where sensory cells containing Arg-Phe-amide-like peptides are located. The external stimuli initiate the release of an internal signal, which is transmitted to the posterior end causing the successive transformation of larval into adult tissue. The transformation front moves from the anterior to the posterior quarter in 60 min. The internal signal can be released or bypassed by a transitory lowering of the Mg2+ content of the seawater. By using this procedure, or by administering an extract containing the putative internal signal substance, each isolated part of the larva can be induced to metamorphose separately. Provided there is no time for regeneration after cutting before metamorphosis is initiated, the most anterior fragment forms only stolons, the most posterior fragment forms only a head. The overt pattern of the polyp is, therefore, generated under the influence of a covert anterior-posterior prepattern of the larva.


Primary malignant melanoma of the transverse colon: report of a case and review of the literature.
Gastrointestinal involvement by malignant melanoma is predominantly a metastatic phenomenon. Although primary malignant melanoma of the gastrointestinal tract has been documented in the esophagus, stomach, small bowel, and anorectum, the incidence of primary melanoma of the colon is rare and remains controversial in most cases. We present a case of solitary malignant melanoma of the transverse colon occurring in a 64-year-old African American male patient. Complete dermatologic and ophthalmologic examinations revealed no evidence of a cutaneous or an ocular primary lesion. Microscopic examination of the resection specimen revealed malignant melanoma, which was confirmed by immunohistochemical positivity for S100 and melan-A. We believe that this tumor represents a primary colonic malignant melanoma.


Purification of antibodies from protein mixtures and mouse ascites fluid using Zeolite X.
Zeolite A and calcium phosphate modified Zeolite A have been shown to be a new effective packing material in ion exchange chromatography for the purification of immunoglobulin G (IgG) from binary mixtures and mouse ascites fluid. This study was to determine the effectiveness of purifying IgG using Zeolite X and dealuminated Zeolite X with twice the pore size of Zeolite A. Binary mixtures (IgG-albumin and IgG-transferrin) and a mouse ascites fluid were purified in Zeolite X (in Na+, K+, or NH4+ form) chromatographic columns and with dealuminated Zeolite X under a variety of operational conditions. The biological activity of the purified IgG from the mouse ascites fluid was confirmed by ELISA. The characteristics of zeolites in the present study suggest that functional groups of a protein displace the cations of zeolites near the crystal surfaces and create a different strength of affinity. The study demonstrated that Zeolite X and dealuminated Zeolite X are also promising new packing materials for the purification of IgG from biological materials.


Endothelin A receptor blockade decreases expression of growth factors and collagen and improves matrix metalloproteinase-2 activity in kidneys from stroke-prone spontaneously hypertensive rats.
This study hypothesizes that endothelin-1 induces renal damage by increasing expression of growth/inflammatory factors, important in renal fibrosis. Male stroke-prone spontaneously hypertensive rats (SHRSPs) (8-weeks, n = 24) were randomized into three groups: control group, high-salt group (4% NaCl), and salt plus an endothelin A receptor antagonist, BMS 182874 (40 mg/kg/d). After 20 weeks treatment, rats were killed. Messenger RNA (mRNA) expression of renal preproendothelin-1, endothelin A and B receptors, and procollagen I and III was evaluated by reverse transcription polymerase chain reaction. Expression of transforming growth factor (TGF)-beta1 and basic fibroblast growth factor (bFGF) was determined by immunoblotting. Matrix metalloproteinase-2 (MMP-2) activity was measured by zymography. In salt-loaded SHRSPs, preproendothelin-1 mRNA expression was increased 1.6-fold, and endothelin A receptor mRNA expression was decreased (70% of control). Salt-loaded SHRSPs had increased renal expression of TGF-b1 and procollagens. MMP-2 activity was augmented fivefold. BMS decreased (p < 0.01) expression of TGF-beta1, bFGF, and procollagen I and reduced MMP-2 activity. Thus severe hypertension and renal dysfunction in salt-loaded SHRSPs are associated with increased expression of renal endothelin-1, growth factors, and collagen. BMS treatment alleviated these effects, suggesting that nephroprotection by endothelin A receptor blockade is mediated by normalizing expression of growth factors, reducing extracellular matrix deposition, and decreasing MMP activity.


A targeted-replacement system for identification of signals for de novo methylation in Neurospora crassa.
Transformation of eukaryotic cells can be used to test potential signals for DNA methylation. This approach is not always reliable, however, because of chromosomal position effects and because integration of multiple and/or rearranged copies of transforming DNA can influence DNA methylation. We developed a robust system to evaluate the potential of DNA fragments to function as signals for de novo methylation in Neurospora crassa. The requirements of the system were (i) a location in the N. crassa genome that becomes methylated only in the presence of a bona fide methylation signal and (ii) an efficient gene replacement protocol. We report here that the am locus fulfills these requirements, and we demonstrate its utility with the identification of a 2.7-kb fragment from the psi 63 locus as a new portable signal for de novo methylation.


Peer Incivility Among Prelicensure Nursing Students: A Call to Action for Nursing Faculty.
This article reports findings from a study that examined the prevalence of peer incivility among nursing students and the impact of incivility on students' physical and mental health and their perceived stress level. Students who experienced high levels of peer incivility had lower mental health scores (P = .0002), lower physical health scores (P = .0069), and higher stress levels (P = .0016). Students who experienced incivility in the classroom had higher levels of peer incivility (P < .001) and lower physical health scores (P < .05).


Photoredox Deaminative Alkylation in DNA-Encoded Library Synthesis.
Herein, we report an on-DNA photoredox-mediated deaminative alkylation method for diversifying DNA-tagged acrylamide substrate with amine-derived radicals. The radicals can be conveniently generated from sterically hindered primary amines, and the deaminative alkylation can tolerate a broad array of radical precursors. Furthermore, the methodology is applicable to Boc-protected diamines, free amino acids, and aryl halides, which bear functional groups enabling additional rounds of diversification. The method is believed to offer a high potential for constructing DNA-encoded libraries, as was demonstrated by the production of a mock library in a 2 × 3 matrix format and confirmation of DNA stability by UPLC-MS and qPCR experiments.


A specific CD4 epitope bound by tregalizumab mediates activation of regulatory T cells by a unique signaling pathway.
CD4(+)CD25(+) regulatory T cells (Tregs) represent a specialized subpopulation of T cells, which are essential for maintaining peripheral tolerance and preventing autoimmunity. The immunomodulatory effects of Tregs depend on their activation status. Here we show that, in contrast to conventional anti-CD4 monoclonal antibodies (mAbs), the humanized CD4-specific monoclonal antibody tregalizumab (BT-061) is able to selectively activate the suppressive properties of Tregs in vitro. BT-061 activates Tregs by binding to CD4 and activation of signaling downstream pathways. The specific functionality of BT-061 may be explained by the recognition of a unique, conformational epitope on domain 2 of the CD4 molecule that is not recognized by other anti-CD4 mAbs. We found that, due to this special epitope binding, BT-061 induces a unique phosphorylation of T-cell receptor complex-associated signaling molecules. This is sufficient to activate the function of Tregs without activating effector T cells. Furthermore, BT-061 does not induce the release of pro-inflammatory cytokines. These results demonstrate that BT-061 stimulation via the CD4 receptor is able to induce T-cell receptor-independent activation of Tregs. Selective activation of Tregs via CD4 is a promising approach for the treatment of autoimmune diseases where insufficient Treg activity has been described. Clinical investigation of this new approach is currently ongoing.


Resistance to thyroid hormone caused by two mutant thyroid hormone receptors beta, R243Q and R243W, with marked impairment of function that cannot be explained by altered in vitro 3,5,3'-triiodothyroinine binding affinity.
Resistance to thyroid hormone (RTH) is a syndrome of reduced responsiveness to thyroid hormone caused by mutations in the thyroid hormone receptor beta (TRbeta) gene. Mutant TRbetas exhibit variable degrees of impaired T3 binding resulting in reduced T3-mediated function. The dominant mode of inheritance is attributed to the ability of mutant TRbetas to interfere with the function of the wild-type (WT) TR, a phenomenon known as dominant negative effect (DNE). We recently identified two families with RTH having mutations in amino acid 243 (R243Q and R243W) in whom the mechanism of RTH appears to be distinct from that of other natural TRbeta mutations. These mutations, which are located in the hinge domain of the TRbeta, do not significantly alter the binding affinity for T3, measured in vitro. The present study was undertaken to characterize the properties of these mutant TRbetas to understand the molecular basis of the RTH phenotype. Two other mutant TRbeta producing RTH with mild (320H) and severe (345R) impairment of T3 binding were studied in parallel. The results demonstrate that TRbetas 243Q and 243W could be translocated into the nucleus where they exerted normal ligand-independent repression of positively regulated thyroid hormone response elements. Yet, the addition of 10 nmol/L T3 failed to normalize the transactivation (16-13% of WT) and revert the DNE exerted by the two TRbeta mutants. In contrast, at this T3 concentration, the transactivation function of 320H was significantly higher (50% of WT), and the DNE was completely abolished, in keeping with the mild clinical form of RTH. Formation of 243Q and 243W homodimers on thyroid hormone response elements could not be as readily prevented by T3 as those formed by the WT and 320H TRbetas. These results suggest that the substitution of R243 in TRbeta produces RTH by increasing the propensity for the formation of tightly bound homodimers or by reduction of the receptor affinity for T3 only after it binds to DNA.


Mettl3-catalyzed m6A regulates histone modifier and modification expression in self-renewing somatic tissue.
N6-methyladenosine (m6A) is the most abundant modification on messenger RNAs (mRNAs) and is catalyzed by methyltransferase-like protein 3 (Mettl3). To understand the role of m6A in a self-renewing somatic tissue, we deleted Mettl3 in epidermal progenitors in vivo. Mice lacking Mettl3 demonstrate marked features of dysfunctional development and self-renewal, including a loss of hair follicle morphogenesis and impaired cell adhesion and polarity associated with oral ulcerations. We show that Mettl3 promotes the m6A-mediated degradation of mRNAs encoding critical histone modifying enzymes. Depletion of Mettl3 results in the loss of m6A on these mRNAs and increases their expression and associated modifications, resulting in widespread gene expression abnormalities that mirror the gross phenotypic abnormalities. Collectively, these results have identified an additional layer of gene regulation within epithelial tissues, revealing an essential role for m6A in the regulation of chromatin modifiers, and underscoring a critical role for Mettl3-catalyzed m6A in proper epithelial development and self-renewal.


Low thyroid hormones and respiratory-distress syndrome of the newborn. Studies on cord blood.
To investigate the relation between thyroid function and respiratory-distress syndrome we determined thyroid hormone in cord serum from 39 term and 120 premature newborns. In groups matched for gestational age (33 to 37 weeks) and body length, after exclusion of newborns with birth asphyxia, maternal diabetes and delivery by cesarean section, the newborns with respiratory distress syndrome had significantly lower cord tri-iodothyronine index, higher ratio of thyroxine to tri-iodothyronine and higher thyrotropin concentration than those without syndrome (P less than 0.05 or less). Conversely, the frequency and severity of respiratory-distress syndrome were higher in newborns who had low cord tri-iodothyronine and thyroxine concentration, low free tri-iodothyronine index and high thyrotropin levels. These observations show an association between low thyroid activity at birth and respiratory-distress syndrome and are in accord with animal studies showing accelerated lung maturation due to thyroxine administration.


Mechanisms for the antihyperglycemic effect of sitagliptin in patients with type 2 diabetes.
Dipeptidyl peptidase IV (DPP-4) inhibitors improve glycemic control in patients with type 2 diabetes. The underlying mechanisms (incretin effect, β-cell function, endogenous glucose production) are not well known. The aim of the study was to examine mechanisms of the antihyperglycemic effect of DPP-4 inhibitors. We administered a mixed meal with glucose tracers ([6,6-(2)H(2)]-glucose infused, [1-(2)H]-glucose ingested), and on a separate day, a glucose infusion matched the glucose responses to the meal (isoglycemic test) in 50 type 2 diabetes patients (hemoglobin A(1c) = 7.4 ± 0.8%) and seven controls; 47 diabetic completers were restudied after 6 wk. Glucose fluxes were calculated, and β-cell function was assessed by mathematical modeling. The incretin effect was calculated as the ratio of oral to iv insulin secretion. We conducted a 6-wk, double-blind, randomized treatment with sitagliptin (100 mg/d; n = 25) or placebo (n = 22). Relative to placebo, meal-induced changes in fasting glucose and glucose area under the curve (AUC) were greater with sitagliptin, in parallel with a lower appearance of oral glucose [difference (post-pre) AUC = -353 ± 915 vs. +146 ± 601 μmol · kg(-1) · 5 h] and greater suppression of endogenous glucose production. Insulin sensitivity improved 10%, whereas total insulin secretion was unchanged. During the meal, β-cell glucose sensitivity improved (+19[29] vs. 5[21] pmol · min(-1) · m(-2) · mm(-1); median [interquartile range]) and glucagon AUC decreased (19.6 ± 7.5 to 17.3 ± 7.1 ng · ml(-1) · 5 h), whereas intact glucose-dependent insulinotropic polypeptide and glucagon-like peptide-1 AUC increased with sitagliptin vs. placebo. The incretin effect was unchanged because sitagliptin increased β-cell glucose sensitivity also during the isoglycemic test. Chronic sitagliptin treatment improves glycemic control by lowering the appearance of oral glucose, postprandial endogenous glucose release, and glucagon response, and by improving insulin sensitivity and β-cell glucose sensing in response to both oral and iv glucose.


A Novel Frameshift Variant of the MITF Gene in a Chinese Family with Waardenburg Syndrome Type 2.
Waardenburg syndrome (WS) is a rare genetic disorder characterized by varying combinations of sensorineural hearing loss and abnormal pigmentation involving the hair, skin and iris. WS is classified into 4 subtypes (WS1-WS4) based on additional symptoms. WS2 is characterized by the absence of additional symptoms and is mainly attributed to variants in the microphthalmia-associated transcription factor (MITF) gene. We detected a novel frameshift variant c.1025_1032delGGAACAAG (NM_198159) of MITF in 5 patients with WS2 from the same Chinese family by using targeted next-generation sequencing and Sanger sequencing. Phenotypic and genotypic analyses of the family members suggested that this novel variants was pathogenic. Our finding expands the spectrum of MITF variants.


Assessment of corrosion in retrieved spine implants.
Recently the use of dissimilar metals in spine instrumentation has increased, especially in the case of adult deformities, where rods made from Cobalt Chrome alloys (CoCr) are used with Titanium (Ti) screws. The use of dissimilar metals increases the risk of galvanic corrosion and patients have required revision spine surgery due to severe metallosis that may have been caused by corrosion. We aimed to assess the presence of corrosion in spine implant retrievals from constructs with two types of material combinations: similar (Ti/Ti) and dissimilar (CoCr/Ti). First, we devised a grading score for corrosion of the rod-fixture junctions. Then, we applied this score to a collection of retrieved spine implants. Our proposed corrosion grading score was proven reliable (kappa > 0.7). We found no significant difference in the scores between 4 CoCr and 11 Ti rods (p = 0.0642). There was no indication that time of implantation had an effect on the corrosion score (p = 0.9361). We recommend surgeons avoid using implants designs with dissimilar metals to reduce the risk of corrosion whilst a larger scale study of retrieved spine implants is conducted. Future studies can now use our scoring system for spine implant corrosion. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 632-638, 2018.


Antioxidant and Anti-Inflammatory Activities of Agrimonia pilosa Ledeb. Extract.
The purpose of this study is to investigate the effect of Agrimonia pilosa Ledeb. extract (APLE) on lipopolysaccharide- (LPS-) induced cell damage in hepatocytes with a focus on antioxidant and anti-inflammatory activities. Total antioxidant and anti-inflammatory activities of APLE itself were analyzed and phytochemical analysis was performed. Moreover, inhibitory effects of APLE on LPS-induced oxidative stress and inflammation were assessed in human HepG2 hepatocytes. APLE was found to exert α,α-diphenyl-β-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and nitrite scavenging activities and reducing power in a dose-dependent manner. The total phenolic and flavonoid contents of APLE were 44.30 ± 1.61 mg GAE/g and 29.65 ± 1.81 mg QE/g, respectively. HPLC analysis revealed that gallic acid is the major phenolic compound in APLE, followed by rutin, genistein, taxifolin, quercetin, luteolin, and apigenin, in descending order. Treatment of 100 and 200 μg/mL APLE significantly reduced LPS-stimulated intracellular reactive oxygen species production to the basal level without any cytotoxicity. Oppositely, APLE reversed LPS-suppressed expression of glutathione peroxidase gene and protein. Consistent with this result, APLE suppressed LPS-triggered expression of proinflammatory cytokine genes in a dose-dependent manner. These results reinforce the fact that the antioxidant and anti-inflammatory activity of APLE helps protect hepatocytes from LPS. Thus, APLE may be utilized as a bioactive ingredient in functional foods.


[To the issue of local treatment in combination therapy of chronic urethritis, associated with sexually transmitted infections].
The article shows the high efficacy of the additional local use of the drug Miramistin in combination therapy of chronic urethritis, associated with sexually transmitted infections (STIs). In accordance with the principles of evidence-based medicine, patients were assigned to the study group (n=110) treated with conventional therapy and Miramistin, and the comparison group (n=40) treated with conventional therapy only. The between-group comparison of treatment effectiveness was carried out by matching results of the etiological healing, the changes of the endoscopic picture of the urethra, and basic clinical manifestations of STI: the degree of inflammatory reaction of urethral mucous membrane, dysuria, pain and sexual syndrome.


Transcriptional induction of cyclooxygenase-2 in osteoblasts is involved in interleukin-6-induced osteoclast formation.
Interleukin-6 (IL-6) induces osteoclast-like cell (osteoclast) formation in a dose-dependent fashion in cocultures of mouse bone marrow cells and osteoblastic cells when soluble IL-6 receptor (sIL-6R) is present. Simultaneous treatment with submaximal doses of IL-1alpha and IL-6 with sIL-6R caused marked induction of osteoclast formation and PGE2 synthesis. These effects were suppressed by adding neutralizing antibodies against IL-1alpha or IL-6R and were totally abolished by adding nonsteroidal antiinflammatory drugs, such as indomethacin and a selective cyclooxygenase-2 (COX-2) inhibitor (NS398). In mouse osteoblastic cells, both IL-1alpha and IL-6 with sIL-6R markedly induced messenger RNA expression of COX-2, but not COX-1, as determined by Northern blot analysis, and luciferase activity in cells stably transfected with a COX-2 promoter-luciferase fusion construct. IL-6 and sIL-6R, when added separately, did not stimulate COX-2 messenger RNA expression. Simultaneous addition of IL-1alpha and IL-6 with sIL-6R to osteoblast cultures cooperatively induced transcription of COX-2, which was associated with a marked increase in COX activity measured by the conversion of arachidonic acid into PGE2. The increased PGE2 synthesis by osteoblasts may play an important role in osteoclastogenesis induced by submaximal doses of IL-1 and IL-6.


A meta-analysis of coffee consumption and pancreatic cancer.
Since when in 1981 a case-control study showed a positive association between coffee and pancreatic cancer, several studies reported inconsistent results on this issue. We conducted a systematic bibliography search updated March 2011 to identify observational studies providing quantitative estimates for pancreatic cancer risk in relation to coffee consumption. We used a meta-analytic approach to estimate overall relative risk (RR) and 95% confidence interval (CI) for the highest versus the lowest coffee consumption categories, using random-effects models. Based on 37 case-control and 17 cohort studies (10,594 cases), the pooled RR for the highest versus lowest intake was 1.13 (95% CI 0.99-1.29). Considering only the smoking-adjusting studies, the pooled RRs were 1.10 (95% CI 0.92-1.31) for the 22 case-control, 1.04 (95% CI 0.80-1.36) for the 15 cohort, and 1.08 (95% CI 0.94-1.25) for all studies. The pooled RR for the increment of one cup of coffee per day was 1.03 (95% CI 0.99-1.06) for the 28 smoking-adjusting studies reporting three or more coffee consumption categories. No significant heterogeneity was observed across strata of study design, sex, geographic region, and other selected characteristics. This meta-analysis provides quantitative evidence that coffee consumption is not appreciably related to pancreatic cancer risk, even at high intakes.


Two minuses can make a plus: waterlogging and elevated CO2 interactions in sweet cherry (Prunus avium) cultivars.
The increase in the ambient concentration of CO2 and other greenhouse gases is producing climate events that can compromise crop survival. However, high CO2 concentrations are sometimes able to mitigate certain stresses such as salinity or drought. In this experiment, the effects of waterlogging and CO2 are studied in combination to elucidate the eventual response in sweet cherry trees. For this purpose, four sweet cherry cultivars ('Burlat', 'Cashmere', 'Lapins and 'New Star') were grafted on a typically hypoxia-tolerant rootstock (Mariana 2624) and submitted to waterlogging for 7 days at either ambient CO2 concentration (400 µmol mol-1 ) or at elevated CO2 (800 µmol mol-1 ). Waterlogging affected plants drastically, by decreasing photosynthesis, stomatal conductance, transpiration, chlorophyll fluorescence and growth. It also brought about the accumulation of proline, chloride and sulfate. Nonetheless, raising the CO2 supply not only mitigated all these effects but also induced the accumulation of soluble sugars and starch in the leaf. Therefore, sweet cherry plants submitted to waterlogging were able to overcome this stress when grown in a CO2 -enriched environment.


Nest architecture is linked with ecological success in songbirds.
Nests are essential constructions that determine fitness, yet their structure can vary substantially across bird species. While there is evidence supporting a link between nest architecture and the habitat a species occupies, we still ignore what ecological and evolutionary processes are linked to different nest types. Using information on 3175 species of songbirds, we show that-after controlling for latitude and body size-species that build domed nests (i.e. nests with a roof) have smaller ranges, are less likely to colonise urban environments and have potentially higher extinction rates compared to species with open and cavity nests. Domed nests could be a costly specialisation, and we show that these nests take more time to be built, which could restrict breeding opportunities. These diverse strands of evidence suggest that the transition from domed to open nests in passerines could represent an important evolutionary innovation behind the success of the largest bird radiation.