Effects of sevoflurane anesthesia on cognitive function and cholinergic signaling

In recent years, concerns have been raised regarding the potential negative impact of anesthetic drugs in postoperative cognitive dysfunction. This thesis is an attempt to contribute to the understanding of the underlying mechanisms. Previous animal studies have suggested that volatile anesthetics can cause long-lasting cognitive alterations in various settings. Based on the strong association between cholinergic function and cognitive performance, we have focused our studies on the long-term effects of volatile anesthetics on cholinergic neurotransmission. By using a combination of laboratory animal experiments and molecular biology techniques in cultured cells, we have described effects of sevoflurane anesthesia on cognitive function and cholinergic signaling. Our studies on spontaneous exploratory behavior, anxiety, object memory and analyses of behavioral organization in mice show that for some aspects of cognitive performance sevoflurane can cause long-lasting effects, while other aspects are unaltered by anesthesia. We have shown that age and preexisting cholinergic dysfunction are factors that influence the results. A normal cholinergic system, as represented by young wild type mice, seems to protect from behavioral alterations after anesthesia. Our results suggest that sevoflurane causes a sustained attenuation in acetylcholine-induced phosphorylation of important intracellular kinases. We have linked our findings to signaling via muscarinic acetylcholine receptors, and we have also demonstrated that the effect of sevoflurane is not likely exerted by reducing the receptor population on the cell surface, nor by altering the total amount of receptor protein available for ligand binding. The overall conclusion from the results of this thesis is that under certain experimental conditions, sevoflurane induces long-lasting alterations both in animal behavior and in cellular signaling

Droppbevattningens inverkan på kvalitet och kvantitet hos Solanum tuberosum L i jämförelse med konventionell spridarebevattning

A literature study and a field trial have been carried out in order to investigate the physiological response of potato plants to different soil moisture levels. Higher yield, quality and number of tubers can according to the literature be obtained if constant high soil moisture is kept during major parts of the growing season. A field trial was set up where drip irrigation was used as an alternative to conventional irrigation techniques to maintain high and constant soil moisture. The application of water in the drip irrigated area was monitored and adjusted on a daily basis so that the soil moisture stayed as close as possible to the recommended level. The sprinkle irrigated area was irrigated according to traditional practices with approximately 35 mm every 7th to 10th day depending on the whether. The result from the field trial was in line with the information found in literature; constant high soil moisture gave on average 11 % higher yield, higher quality and higher number of tubers. The marketable yield increased with 28 % in the drip irrigated area compared with the sprinkle irrigated area. The conclusion from this study is that drip irrigation can be used as a cost effective alternative to traditional sprinkle irrigation.En litteraturstudie samt ett fältförsök har genomförts för att studera markfuktens inflytande på kvalitet och kvantitet hos Solanum tuberosum. Litteraturstudien visar att hög och jämn markfukt vid odling av potatis ger högre skörd, fler antal knölar och bättre kvalitet. Ett praktiskt odlingsförsök genomfördes för att undersöka om teorin överenstämmer med praktiken. I försöket testades droppbevattning som ett alternativ till konventionell spridarbevattning i ett enkelt blockförsök. Markfukten i det droppbevattnade ledet styrdes utifrån avdustningen och reglerades så att den höll sig inom det rekomenderade intervallet. Det spridarbevattnade ledet vattnades på traditionsenligt vis med ca 35 mm var 7-10 dag beroende på väderlek. Resultaten från försöket styrkte teorin, en hög och jämn markfukt gav i medeltal 11 % högre skörd och den säljbara fraktionen, dvs. storleken mellan 38 och 55mm, blev 20 % högre. Största ökningen blev det i fraktionen 38-45 mm där det droppbevattnade ledet gav 36 % mer skörd än det spridarbevattnade. I medeltal gav det droppbevattnade ledet 62 % mindre potatis i fraktionen större än 55 mm. Andelen foder i det droppbevattnade ledet minskade så att den totala säljbara varan ökade med i genomsnitt 28 % i jämförelse med det spridarbevattnade ledet. Slutsatsen från denna studie är att droppbevattning kan användas som ett kostnadseffektivt alternativ till spridarbevattning

Influence of consecutive-day blood sampling on polymerase chain reaction-adjusted parasitological cure rates in an antimalarial-drug trial conducted in Tanzania

We assessed the influence that consecutive-day blood sampling, compared with single-day blood sampling, had on polymerase chain reaction (PCR)-adjusted parasitological cure after stepwise genotyping of merozoite surface proteins 2 (msp2) and 1 (msp1) in 106 children in Tanzania who had uncomplicated falciparum malaria treated with either sulfadoxine-pyrimethamine or artemether- lumefantrine; 78 of these children developed recurrent parasitemia during the 42-day follow-up period. Initial msp2 genotyping identified 27 and 33 recrudescences by use of single- and consecutive-day sampling, respectively; in subsequent msp1 genotyping, 17 and 21 of these episodes, respectively, were still classified as recrudescences; these results indicate a similar sensitivity of the standard single-day PCR protocol - that is, 82% (27/33) and 81% (17/21), in both genotyping steps. Interpretation of PCR-adjusted results will significantly depend on methodology. © 2007 by the Infectious Diseases Society of America. All rights reserved

Multilocus sequence analysis reveals different lineages of Pseudomonas anguilliseptica associated with disease in farmed lumpfish (Cyclopterus lumpus L.)

The bacterium Pseudomonas anguilliseptica has in recent years emerged as a serious threat to production of lumpfish in Norway. Little is known about the population structure of this bacterium despite its association with disease in a wide range of different fish species throughout the world. The phylogenetic relationships between 53 isolates, primarily derived from diseased lumpfish, but including a number of reference strains from diverse geographical origins and fish species, were reconstructed by Multi-Locus Sequence Analysis (MLSA) using nine housekeeping genes (rpoB, atpD, gyrB, rpoD, ileS, aroE, carA, glnS and recA). MLSA revealed a high degree of relatedness between the studied isolates, altough the seven genotypes identified formed three main phylogenetic lineages. While four genotypes were identified amongst Norwegian lumpfish isolates, a single genotype dominated, irrespective of geographic origin. This suggests the existence of a dominant genotype associated with disease in production of lumpfish in Norwegian aquaculture. Elucidation of the population structure of the bacterium has provided valuable information for potential future vaccine development.publishedVersio

Plasmodium falciparum multidrug resistance protein 1 and artemisinin-based combination therapy in Africa

Plasmodium falciparum response mechanisms to the major artemisinin-based combination therapies (ACTs) are largely unknown. Multidrug-resistance protein (MRP)-like adenosine triphosphate (ATP)-binding cassette transporters are known to be related to multidrug resistance in many organisms. Therefore, we hypothesized that sequence variation in pfmrp1 can contribute to decreased parasite sensitivity to ACT. Through sequencing of the pfmrp1 open reading frame for 103 geographically diverse P. falciparum infections, we identified 27 single-nucleotide polymorphisms (SNPs), of which 21 were nonsynonymous and 6 synonymous. Analyses of clinical efficacy trials with artesunate-amodiaquine and artemether-lumefantrine detected a specific selection of the globally prevalent I876V SNP in recurrent infections after artemether-lumefantrine treatment. Additional in silico studies suggested an influence of variation in amino acid 876 on the ATP hydrolysis cycle of pfMRP1 with potential impact on protein functionality. Our data suggest for the first time, to our knowledge, the involvement of pfMRP1 in P. falciparum in vivo response to ACT.info:eu-repo/semantics/publishedVersio

Screen-printed ultrasonic 2-D matrix array transducers for microparticle manipulation

This paper reports the development of a two-dimensional thick film lead zirconate titanate (PZT) ultrasonic transducer array, operating at frequency approximately 7.5 MHz, to demonstrate the potential of this fabrication technique for microparticle manipulation. All layers of the array are screen-printed then sintered on an alumina substrate without any subsequent patterning processes. The thickness of the thick film PZT is 139 ± 2 μm, the element pitch of the array is 2.3 mm, and the dimension of each individual PZT element is 2 × 2 mm2 with top electrode 1.7 × 1.7 mm2. The measured relative dielectric constant of the PZT is 2250 ± 100 and the dielectric loss is 0.09 ± 0.005 at 10 kHz. Finite element analysis was used to predict the behaviour of the array and to optimise its configuration. Electrical impedance spectroscopy and laser vibrometry were used to characterise the array experimentally. The measured surface motion of a single element is on the order of tens of nanometres with a 10 Vpeak continuous sinusoidal excitation. Particle manipulation experiments have been demonstrated with the array by manipulating Ø10 μm polystyrene microspheres in degassed water. The simplified array fabrication process and the bulk production capability of screen-printing suggest potential for the commercialisation of multilayer planar resonant devices for ultrasonic particle manipulation

Neuroinflammatory markers associate with cognitive decline after major surgery:Findings of an explorative study

OBJECTIVE Long‐term cognitive decline is an adverse outcome after major surgery associated with increased risk for mortality and morbidity. We studied the cerebrospinal fluid (CSF) and serum biochemical inflammatory response to a standardized orthopedic surgical procedure and the possible association with long‐term changes in cognitive function. We hypothesized that the CSF inflammatory response pattern after surgery would differ in patients having long‐term cognitive decline defined as a composite cognitive z score of ≥1.0 compared to patients without long‐term cognitive decline at 3 months postsurgery. METHODS Serum and CSF biomarkers of inflammation and blood–brain barrier (BBB) integrity were measured preoperatively and up to 48 hours postoperatively, and cognitive function was assessed preoperatively and at 2 to 5 days and 3 months postoperatively. RESULTS Surgery was associated with a pronounced increase in inflammatory biomarkers in both CSF and blood throughout the 48‐hour study period. A principal component (PC) analysis was performed on 52 inflammatory biomarkers. The 2 first PC (PC1 and PC2) construct outcome variables on CSF biomarkers were significantly associated with long‐term cognitive decline at 3 months, but none of the PC construct serum variables showed a significant association with long‐term cognitive decline at 3 months. Patients both with and patients without long‐term cognitive decline showed early transient increases of the astroglial biomarkers S‐100B and glial fibrillary acidic protein in CSF, and in BBB permeability (CSF/serum albumin ratio). INTERPRETATION Surgery rapidly triggers a temporal neuroinflammatory response closely associated with long‐term cognitive outcome postsurgery. The findings of this explorative study require validation in a larger surgical patient cohort. ANN NEUROL 202

Optimization and validation of multi-coloured capillary electrophoresis for genotyping of Plasmodium falciparum merozoite surface proteins (msp1 and 2)

BACKGROUND: Genotyping of Plasmodium falciparum based on PCR amplification of the polymorphic genes encoding the merozoite surface proteins 1 and 2 (msp1 and msp2) is well established in the field of malaria research to determine the number and types of concurrent clones in an infection. Genotyping is regarded essential in anti-malarial drug trials to define treatment outcome, by distinguishing recrudescent parasites from new infections. Because of the limitations in specificity and resolution of gel electrophoresis used for fragment analysis in most genotyping assays it became necessary to improve the methodology. An alternative technique for fragment analysis is capillary electrophoresis (CE) performed using automated DNA sequencers. Here, one of the most widely-used protocols for genotyping of P. falciparum msp1 and msp2 has been adapted to the CE technique. The protocol and optimization process as well as the potentials and limitations of the technique in molecular epidemiology studies and anti-malarial drug trials are reported. METHODS: The original genotyping assay was adapted by fluorescent labeling of the msp1 and msp2 allelic type specific primers in the nested PCR and analysis of the final PCR products in a DNA sequencer. A substantial optimization of the fluorescent assay was performed. The CE method was validated using known mixtures of laboratory lines and field samples from Ghana and Tanzania, and compared to the original PCR assay with gel electrophoresis. RESULTS: The CE-based method showed high precision and reproducibility in determining fragment size (< 1 bp). More genotypes were detected in mixtures of laboratory lines and blood samples from malaria infected children, compared to gel electrophoresis. The capacity to distinguish recrudescent parasites from new infections in an anti-malarial drug trial was similar by both methods, resulting in the same outcome classification, however with more precise determination by CE. CONCLUSION: The improved resolution and reproducibility of CE in fragment sizing allows for comparison of alleles between separate runs and determination of allele frequencies in a population. The more detailed characterization of individual msp1 and msp2 genotypes may contribute to improved assessments in anti-malarial drug trials and to a further understanding of the molecular epidemiology of these polymorphic P. falciparum antigens

Hundreds of variants clustered in genomic loci and biological pathways affect human height

Most common human traits and diseases have a polygenic pattern of inheritance: DNA sequence variants at many genetic loci influence the phenotype. Genome-wide association (GWA) studies have identified more than 600 variants associated with human traits, but these typically explain small fractions of phenotypic variation, raising questions about the use of further studies. Here, using 183,727 individuals, we show that hundreds of genetic variants, in at least 180 loci, influence adult height, a highly heritable and classic polygenic trait. The large number of loci reveals patterns with important implications for genetic studies of common human diseases and traits. First, the 180 loci are not random, but instead are enriched for genes that are connected in biological pathways (P = 0.016) and that underlie skeletal growth defects (P < 0.001). Second, the likely causal gene is often located near the most strongly associated variant: in 13 of 21 loci containing a known skeletal growth gene, that gene was closest to the associated variant. Third, at least 19 loci have multiple independently associated variants, suggesting that allelic heterogeneity is a frequent feature of polygenic traits, that comprehensive explorations of already-discovered loci should discover additional variants and that an appreciable fraction of associated loci may have been identified. Fourth, associated variants are enriched for likely functional effects on genes, being over-represented among variants that alter amino-acid structure of proteins and expression levels of nearby genes. Our data explain approximately 10% of the phenotypic variation in height, and we estimate that unidentified common variants of similar effect sizes would increase this figure to approximately 16% of phenotypic variation (approximately 20% of heritable variation). Although additional approaches are needed to dissect the genetic architecture of polygenic human traits fully, our findings indicate that GWA studies can identify large numbers of loci that implicate biologically relevant genes and pathways.

Polygenic hazard score to guide screening for aggressive prostate cancer: development and validation in large scale cohorts.

OBJECTIVES: To develop and validate a genetic tool to predict age of onset of aggressive prostate cancer (PCa) and to guide decisions of who to screen and at what age. DESIGN: Analysis of genotype, PCa status, and age to select single nucleotide polymorphisms (SNPs) associated with diagnosis. These polymorphisms were incorporated into a survival analysis to estimate their effects on age at diagnosis of aggressive PCa (that is, not eligible for surveillance according to National Comprehensive Cancer Network guidelines; any of Gleason score ≥7, stage T3-T4, PSA (prostate specific antigen) concentration ≥10 ng/L, nodal metastasis, distant metastasis). The resulting polygenic hazard score is an assessment of individual genetic risk. The final model was applied to an independent dataset containing genotype and PSA screening data. The hazard score was calculated for these men to test prediction of survival free from PCa. SETTING: Multiple institutions that were members of international PRACTICAL consortium. PARTICIPANTS: All consortium participants of European ancestry with known age, PCa status, and quality assured custom (iCOGS) array genotype data. The development dataset comprised 31 747 men; the validation dataset comprised 6411 men. MAIN OUTCOME MEASURES: Prediction with hazard score of age of onset of aggressive cancer in validation set. RESULTS: In the independent validation set, the hazard score calculated from 54 single nucleotide polymorphisms was a highly significant predictor of age at diagnosis of aggressive cancer (z=11.2, P98th centile) were compared with those with average scores (30th-70th centile), the hazard ratio for aggressive cancer was 2.9 (95% confidence interval 2.4 to 3.4). Inclusion of family history in a combined model did not improve prediction of onset of aggressive PCa (P=0.59), and polygenic hazard score performance remained high when family history was accounted for. Additionally, the positive predictive value of PSA screening for aggressive PCa was increased with increasing polygenic hazard score. CONCLUSIONS: Polygenic hazard scores can be used for personalised genetic risk estimates that can predict for age at onset of aggressive PCa