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PubMed2004

Validation and cross-cultural adjustment of PDQL-questionnaire, Spanish version (Ecuador) (PDQL-EV).

OBJECTIVE: To validate the Ecuadorian Spanish-language version of the Parkinson's Disease Quality of Life Questionnaire (PDQL-EV). METHOD: Cross-sectional study of patients with Parkinson's Disease attending a Movement Disorders Clinic. MEASURES APPLIED: Hoehn and Yahr (HY), Unified Parkinson's Disease Rating Scale (UPDRS-3.0), Schwab and England Scale (SES), Pfeiffer Questionnaire (SPMSQ), Hospital Anxiety and Depression Scale (HADS). PDQ-39 and PDQL-EV were applied, the latter after translation and back translation. RESULTS: The study included 137 patients with an age range of 69.43 +/- 10.18 years (mean +/- SD), duration of illness 5.87 +/- 2.58 years, and HY 2.28 +/- 0.77. The PDQL-EV internal consistency (Cronbach's alpha) yielded: Parkinson's symptoms (PS) 0.85; systemic symptoms (SS) 0.69; social function (SF) 0.81; and emotional function (EF) 0.78. The correlation between measurements varied from 0.68 (PS-SS) to 0.46 (SS-EF), and total measurement correlation from 0.58 (EF) to 0.74 (PS) (altogether, p < 0.0001). The correlation of PDQL with clinical variables was: HY -0.62; UPDRS (subscales 1, 2 and 3) -0.55, -0.76, and -0.48, respectively; SES 0.65; SPMSQ -0.35; HADS-anxiety -0.55; HADS-depression -0.67; and PDQ-39 -0.91 (altogether p < 0.0001). PDQL-EV scores were significantly different (p < 0.0001) between moderate and advanced stages, for all domains except for EF (p = 0.006). CONCLUSION: The PDQ-EV has satisfactory internal consistency, and converging and discriminating validity.

Aged↗
PubMed2006

Phosphodiester and N-glycosidic bond cleavage in DNA induced by 4-15 eV electrons.

Thin molecular films of the short single strand of DNA, GCAT, were bombarded under vacuum by electrons with energies between 4 and 15 eV. Ex vacuo analysis by high-pressure liquid chromatography of the samples exposed to the electron beam revealed the formation of a multitude of products. Among these, 12 fragments of GCAT were identified by comparison with reference compounds and their yields were measured as a function of electron energy. For all energies, scission of the backbone gave nonmodified fragments containing a terminal phosphate, with negligible amounts of fragments without the phosphate group. This indicates that phosphodiester bond cleavage by 4-15 eV electrons involves cleavage of the C-O bond rather than the P-O bond. The yield functions exhibit maxima at 6 and 10-12 eV, which are interpreted as due to the formation of transient anions leading to fragmentation. Below 15 eV, these resonances dominate bond dissociation processes. All four nonmodified bases are released from the tetramer, by cleavage of the N-glycosidic bond, which occurs principally via the formation of core-excited resonances located around 6 and 10 eV. The formation of the other nonmodified products leading to cleavage of the phosphodiester bond is suggested to occur principally via two different mechanisms: (1) the formation of a core-excited resonance on the phosphate unit followed by dissociation of the transient anion and (2) dissociation of the CO bond of the phosphate group formed by resonance electron transfer from the bases. In each case, phosphodiester bond cleavage leads chiefly to the formation of stable phosphate anions and sugar radicals with minimal amounts of alkoxyl anions and phosphoryl radicals.

Chromatography, High Pressure Liquid↗
PubMed1998

Transmission multilayer polarizers for use in the 55-90 eV region.

Free-standing Al/YB(6) and Mo/Si transmission multilayer polarizers have been developed for use in the photon energy regions 55-72 eV and 72-90 eV, respectively, to improve the degree of polarization of synchrotron radiation light from beamline monochromators. The polarizance (polarizing ability) and transmittance of the Al/YB(6) multilayer for unpolarized light at 64 eV were 77% and 3.4%, respectively, and the polarizance and transmittance of the Mo/Si multilayer at 80 eV were 79% and 2.2%, respectively. By using the Mo/Si polarizer, the degree of polarization of the monochromated light at BL8B1 of UVSOR was increased from 95 to 99% in the 78-84 eV region.

Journal Article↗
PubMed2001

Low-energy (5-40 eV) electron-stimulated desorption of anions from physisorbed DNA bases.

We present the results of experiments on anion desorption from the physisorbed DNA bases adenine, thymine, guanine and cytosine induced by the impact of low-energy (5-40 eV) electrons. Electron bombardment of DNA base films induces ring fragmentation and desorption of H(-), O(-), OH(-), CN(-), OCN(- ) and CH(2)(-) anions through either single or complex multibond dissociation. We designate the variation of the yield of an anion with electron energy as the yield function. Below 15 eV incident electron energy, bond cleavage is controlled mainly by dissociative electron attachment. Above 15 eV, the portion of a yield function that increases linearly is attributed to nonresonant processes, such as dipolar dissociation. A resonant structure is superimposed on this signal around 20 eV in the anion yield functions. This structure implicates dissociative electron attachment and/or resonant decay of the transient anion into the dipolar dissociation channel, with a minimal contribution from multiple inelastic electron scattering. The yields of all desorbing anions clearly show that electron resonances contribute to the damage of all DNA bases bombarded with 5-40 eV electrons. Comparison of the ion yields indicates that adenine is the least sensitive base to slow electron attack. Electron-irradiated guanine films exhibit the largest yields of desorbed anions.

Adenine↗
PubMed2001

Sequence-specific damage induced by the impact of 3-30 eV electrons on oligonucleotides.

The ability of low-energy electrons to induce single- and double-strand breaks in DNA has recently been demonstrated. Here we show the propensity of 3-30 eV electrons to initiate base sequence-dependent damage to a short single DNA strand. Solid monolayer films of homogeneous thymidine (T(9)) and deoxycytidine (dCy(9)) and heterogeneous oligomers (T(6)dCy(3)) are bombarded with 1-30 eV electrons in an ultrahigh-vacuum system. CN, OCN and/or H(2)NCN are detected by a mass spectrometer as the most intense neutral fragments desorbing in vacuum. A weaker signal of CH(3)CCO is also detected, but only from oligonucleotides containing thymine. Below 17 eV, the energy dependence of the yields of CN, OCN and CH(3)CCO exhibits resonance-like structures, attributed to dissociative electron attachment (DEA). Above 17 eV, the monotonic increase in the fragment yields indicates that nonresonant processes (i.e. dipolar dissociation) control the fragmentation of these molecules. Within the energy range investigated, comparison of the magnitude of the total fragment yields produced by electron attack on dCy(9), T(6)-dCy(3) and T(9) suggests the following order in the sensitivity of single-strand DNA: dCy(9) > T(6)-dCy(3) > T(9). At 12 eV, the total fragment yields are found to be 5.8, 5.0 and 3.9 x 10(-3) fragment/electron, respectively. From the yields obtained with the two homo-oligonucleotides, we differentiate between contributions arising from the chemical nature of the base and the effect of environment (i.e. the sequence) when a thymidine unit in T(9) is replaced by dCy. The base sequence-dependent damage is found to vary with incident electron energy. These results reinforce the idea that genomic sensitivity to ionizing radiation depends on local genetic information. Furthermore, they underscore the possible role of low-energy electrons in the pathways responsible for the induction of specific genomic lesions.

Base Sequence↗
PubMed1986

Effect of esophageal variceal sclerotherapy (EVS) on lung function. A prospective controlled study.

A prospective, controlled study to determine the short- and long-term effects of esophageal variceal sclerotherapy (EVS) on lung function was carried out on 11 patients with cirrhotic portal hypertension and variceal hemorrhage. Eleven patients with chronic liver disease undergoing diagnostic endoscopy served as controls. There was no difference in lung function tests and gas exchange in both the EVS or control groups after either procedure. No change in these parameters was noted during follow-up on continued sclerotherapy in the EVS group. Ventilation-perfusion scans and chest roentgenograms, performed before and after EVS, demonstrated no significant change. We conclude that in patients with stable liver disease and without hepatic failure, EVS does not result in serious short- and long-term impairment of lung function.

Adult↗
PubMed2026

Rapid glycomic analysis of serum EVs reveals altered N-glycosylation patterns in ASD.

Objective laboratory diagnostics for autism spectrum disorder (ASD) are lacking, necessitating rapid clinical screening tools. Because serum extracellular vesicle (EV) N-glycosylation captures critical neurodevelopmental signatures, we developed a fast, biologically interpretable diagnostic strategy. EVs from ASD patients with language impairment and neurotypical controls were isolated using a rapid extra-polyethylene glycol precipitation/filtration (EPF) workflow, benchmarked against ultracentrifugation. Following MALDI-TOF/MS profiling, machine learning was re-evaluated using repeated nested cross-validation to reduce optimistic bias and potential information leakage. Among five classifiers, Random Forest (RF) showed the best overall balance across discrimination, calibration, and classification metrics. RF-based SHAP analysis provided transparent interpretation, highlighting key discriminative glycans, including H4N3S1F1, H5N5S1F1, and H3N5F1. To elucidate molecular mechanisms, we integrated public EV transcriptomic data. This revealed significant dysregulation of N-glycosylation machinery genes (e.g., MAN1A1, NEU1, OSTC, RPN2), whose expression directionally aligned with observed glycan shifts in synaptic pathways. Collectively, this rapid serum EV N-glycomic workflow, combined with leakage-controlled RF-based interpretation, provides a promising foundation for non-invasive ASD biomarker discovery and future multicenter validation.

Humans↗
PubMed1999

An approach to an objective background subtraction for elemental mapping with core-edges down to 50 eV: description, evaluation and application.

To image the distribution of a specific element in a specimen with an energy filtering TEM, the element-unspecific background under the core-edge has to be subtracted. The most commonly used procedure is the three-window power-law method leading to considerable systematic errors for low-energy core-edges. Here a new method is described which can be considered as a generalized difference method. Characteristic examples for element detection in biological specimens using this method are shown. The background under the core-edge can be described by one or two pre-edge windows as a polynome of third order. This function can be deduced from specimen areas that are not known to contain the element or from a second specimen used as a standard. Control experiments showed that background subtraction for on-overlapping core-edges in the low-loss region (50-200 eV) needs two pre-edge images, whereas at higher-energy losses (> 300 eV) only one pre-edge image is necessary. With the method described, objective elemental mapping becomes possible even for edges at 50-100 eV. This was proven for the M2,3-edge of iron at 60 eV. The detection of phosphorous was possible with a signal-to-noise ratio five times higher than when using the three-window method. Preliminary data showed that it should be possible to detect calcium with only one image before the edge.

Animals↗
PubMed2004

Damage to model DNA fragments from very low-energy (<1 eV) electrons.

Although electrons having enough energy to ionize or electronically excite DNA have long been known to cause strand breaks (i.e., bond cleavages), only recently has it been suggested that even lower-energy electrons (most recently 1 eV and below) can also damage DNA. The findings of the present work suggest that, while DNA bases can attach electrons having kinetic energies in the 1 eV range and subsequently undergo phosphate-sugar O-C sigma bond cleavage, it is highly unlikely (in contrast to recent suggestions) that electrons having kinetic energies near 0 eV can attach to the phosphate unit's P=O bonds. Electron kinetic energies in the 2-3 eV range are required to attach directly to DNA's phosphate group's P=O pi orbital and induce phosphate-sugar O-C sigma bond cleavages if the phosphate groups are rendered neutral (e.g., by nearby counterions). Moreover, significant activation barriers to C-O bond breakage render the rates of both such damage mechanisms (i.e., P=O-attached and base-attached) slow as compared to electron autodetachment and to other damage processes.

DNA Damage↗
PubMed2025

EV DNA from pancreatic cancer patient-derived cells harbors molecular, coding, non-coding signatures and mutational hotspots.

DNA packaged into cancer cell-derived EV is not well appreciated. Here, we uncovered signatures of EV DNA secreted by pancreatic cancer cells. The cancer cells and non-cancer counterparts exhibit distinct low vs. high molecular weight (LMW vs. HMW) EV DNA fragments distribution, respectively. Genome sequencing and Single Nucleotide Variants analysis revealed that 95% of reads and 94% of SNVs map to noncoding regions of the genome. Given that ~1% of the human genome represents coding regions, the 5% mapping rate to coding regions suggests a non-random enrichment of certain coding regions and mutations. The LMW DNA fragments not only set cancer cells apart, but also harbor cancer specific enrichment of unique coding regions, the top nine being FAM135B, COL22A1, TSNARE1, KCNK9, ZFAT, JRK, MROH5, GSDMD, and MIR3667HG. Additionally, the cancer cells' LMW DNA fragments exhibit dense centromeric mapping more strikingly on chromosomes 3, 7, 9, 10, 11, 13, 17, and 20. Mutational profiling turned up close to 200 mutations specific for the cancer cells. Altogether, our analyses suggest that centromeric regions might hold clues to EV DNA content from pancreatic cancer, the molecular, mutational signatures thereof, and rationalizes the need for a new approach to DNA biomarker research.

Humans↗
PubMed2006

Induction of strand breaks by low-energy electrons (8-68 eV) in a self-assembled monolayer of oligonucleotides: effective cross sections and attenuation lengths.

Self-assembled monolayers of 5'-32P-labeled 3'-thiolated oligonucleotides chemisorbed on gold were bombarded by low-energy electrons (LEE) of 8-68 eV. Shorter 5'-32P-oligonucleotides produced by LEE-induced strand breaks were separated with denaturing polyacrylamide gel electrophoresis and quantified by phosphor imaging. The yields of short oligonucleotides (y) decrease exponentially with their length (n), following the equation y=ae-bn, where a and b are constants, which are related to the average effective cross section per nucleotide for DNA strand break (sigmaeff) and the attenuation length (AL=1b) of LEE, respectively. The AL decreases with LEE energies from 2.5+/-0.6 nm at 8 eV to 0.8+/-0.1 nm at 68 eV, whereas sigmaeff increases from (3+/-1)x10(-18) to (5.1+/-1.6)x10(-17) cm2 within the same energy range. The energy dependence of sigmaeff shows a resonance peak of (2.8+/-0.9)x10(-17) cm2 at 18 eV superimposed on a monotonically rising curve. Transient electron attachment to a sigma* anion state of the deoxyribose group, followed by dipolar dissociation into H- and the corresponding positive-ion radical, leading to C-O bond cleavage, is proposed to account for this maximum.

Journal Article↗
PubMed2006

Excitation-energy dependence of the mechanism for two-photon ionization of liquid H(2)O and D(2)O from 8.3 to 12.4 eV.

Transient absorption measurements monitor the geminate recombination kinetics of solvated electrons following two-photon ionization of liquid water at several excitation energies in the range from 8.3 to 12.4 eV. Modeling the kinetics of the electron reveals its average ejection length from the hydronium ion and hydroxyl radical counterparts and thus provides insight into the ionization mechanism. The electron ejection length increases monotonically from roughly 0.9 nm at 8.3 eV to nearly 4 nm at 12.4 eV, with the increase taking place most rapidly above 9.5 eV. We connect our results with recent advances in the understanding of the electronic structure of liquid water and discuss the nature of the ionization mechanism as a function of excitation energy. The isotope dependence of the electron ejection length provides additional information about the ionization mechanism. The electron ejection length has a similar energy dependence for two-photon ionization of liquid D(2)O, but is consistently shorter than in H(2)O by about 0.3 nm across the wide range of excitation energies studied.

Journal Article↗
PubMed1980

Chromosomal localization of ev-1, a frequently occurring endogenous retrovirus locus in white Leghorn chickens, by in situ hybridization.

The distribution of 12 endogenous retrovirus loci from 13 White Leghorn flocks of diverse geographical origin was examined, using a total of 508 birds. Only one locus, designated ev-1, was observed to be almost universally present (506 of 508 birds) and thus predominantly homozygous in this breed of chicken. By contrast, 11 other viral loci were observed at much lower frequencies, 3 to 30%, and were occasionally restricted to individual flocks. To further characterize the commonly occurring ev-1 viral locus and to provide a foundation for determining the mechanism by which these 12 endogenous retrovirus loci were generated, a new technique of indirect in situ hybridization was used to localize the chromosomal position of ev-1. By using a combination of specific probes, the position of ev-1 was localized to a small region on the long arm of chromosome 1. This research represents oen of the first instances in which a defined segment of nonreiterated DNA coding for a known set of genes has been localized to a specific region on a vertebrate chromosome by in situ hybridization.

Animals↗
PubMed2025

Proteomic-based identification of novel EV-derived protein antibodies biomarkers for melioidosis diagnosis.

Melioidosis, caused by Burkholderia pseudomallei (Bp), is a life-threatening disease characterized by diverse clinical manifestations and limited diagnostic capabilities. Extracellular vesicles (EVs) have emerged as critical carriers of novel antibody targets for serodiagnosis. In this study, we established a Bp-infected BEAS-2B cell model (Bp/BEAS-2B) and isolated EV from both Bp and Bp/BEAS-2B cells to generate EV proteome, identifying potential antigenic biomarkers for melioidosis diagnosis. Bioinformatics analysis identified PPEP and POMCR proteins as candidate antigens, with BLF1 and omp A serving as positive controls. Using a self-developed IgM-ELISA, serum samples from 43 melioidosis patients and 47 healthy volunteers were analyzed to detect antibodies against these antigens. Anti-POMCR IgM demonstrated exceptional diagnostic performance, with an AUC of 0.9872 (95% CI: 0.9713-1.003), sensitivity of 93.02% and specificity of 97.92% at a cutoff value of OD450&#x2009;=&#x2009;0.118. Similarly, IgM against PPEP, BLF1, and omp A also showed high diagnostic accuracy, with AUC values of 0.969, 0.9621, and 0.976, respectively. The accuracy of anti-POMCR and anti-PPEP were 96.43% and 95.54%, respectively, equivalent to anti-omp A (93.75%) and anti-BLF1 (91.96%). Antibodies to EV-derived proteins effectively differentiated melioidosis patients from other bacterial infections and healthy volunteers, highlighting their clinical potential as diagnostic tools for melioidosis.

Humans↗
PubMed2006

Field efficacy of the BG-Sentinel compared with CDC Backpack Aspirators and CO2-baited EVS traps for collection of adult Aedes aegypti in Cairns, Queensland, Australia.

In this study, we compared the efficacy of the newly available BG-Sentinel with an established "gold standard,", the CDC Backpack Aspirator, and a CO2-baited EVS trap for the collection of Aedes aegypti (L.) in Cairns, Australia. BG-Sentinels collected significantly more (P = 0.017) female Ae. aegypti (mean per collection, 1.92 +/- 0.39) than both the CDC Backpack Aspirator (1.00 +/- 0.35) and the EVS trap (0.71 +/- 0.27). Male-only and combined male-female Ae. aegypti collections for th BG-Sentinel and the CDC Backpack Aspirator were also greater than EVS trap collections. The CDC Backpack Aspirator and the BG-Sentinel captured proportionally fewer females compared with the EVS trap. The BG-Sentinel was the most Ae. aegypti specific collection method. The CDC Backpack Aspirator collected proportionally more bloodfed Ae. aegypti than the other methods, which collected a greater proportion of nullipars. The data presented here will aid researchers in deciding what Ae. aegypti sampling device best suits their needs. BG-Sentinels and CDC Backpack Aspirators should be considered as alternatives to human-bait collections for Ae. aegypti sampling.

Aedes↗
PubMed2004

The relative attractiveness of carbon dioxide and octenol in CDC- and EVS-type light traps for sampling the mosquitoes Aedes aegypti (L.), Aedes polynesiensis Marks, and Culex quinquefasciatus say in Moorea, French Polynesia.

Two dominant day-biting pests and vector species on the island of Moorea in French Polynesia are Aedes (Stegomyia) aegypti (L.) and Aedes (Stegomyia) polynesiensis Marks, major vectors of dengue viruses and Wuchereria bancrofti, respectively. Their surveillance is hindered by a relative lack of attraction to light traps, necessitating the undesirable use of human bait collections with the inherent risks of pathogen transmission. The effectiveness of CDC- and EVS-type light traps baited with olfactory attractants was evaluated for these two Aedes species and the nocturnal Culex (Culex) quinquefasciatus Say in three sites in urban and semi-rural environments on Moorea in October/November 2003. Firstly, four CDC-type traps with light only, light with octenol, light with carbon dioxide (dry ice), and light with octenol plus carbon dioxide were operated continuously over four days with daily rotation to compensate for position effects. Secondly, two CDC- and two EVS-type traps with carbon dioxide or carbon dioxide plus octenol were operated continuously over four days with similar rotation. Variation was found in the numbers of the three species collected at the different sites, reflecting the relative availability of their preferred larval habitats. With the CDC traps in the first trial, the addition of octenol to the light did not significantly increase the collection of any species, the addition of carbon dioxide did significantly increase collection of all three species, while the addition of octenol to the light plus carbon dioxide did not significantly increase the collections further. In the second trial, there was no significant difference in the mean number of Ae. aegypti or Ae. polynesiensis collected in either EVS or CDC traps when baited with carbon dioxide or with octenol added. For Cx. quinquefasciatus, the supplementation with octenol made no significant difference with EVS traps but resulted in significantly reduced collections in CDC traps. Overall, neither trap, however baited, provided large samples when compared with landing/ biting collections at human bait. Only two other species were collected, Culex (Culex) roseni Belkin and Aedes (Aedimorphus) nocturnus (Theobald), the latter being a first record for the island of Moorea and for French Polynesia.

Aedes↗
PubMed1998

Ultrastructural and immunocytochemical study on the infection of enterovirus 71 (EV 71) in rhabdomyosarcoma (RD) cells.

Rhabdomyosarcoma monolayers were inoculated with enterovirus 71 (EV 71) at 73 degrees C, sampled at intervals during the replicative cycle, and examined in thin sections by electron microscopy, using routine and immunoelectronmicroscopy with polyclonal antibodies against EV 71. The location of EV 71 or its precursors was followed during the viral replicative cycle. The earliest samples (3 h postinoculation) showed a cell shape change, from elongated to rounded. At 6 h postinoculation, the presence of early virus-induced vesicles developing within the cytoplasm was pointed out, although no virus particles were observed at these stages. At 12 and 20 h postinoculation, virus particles appeared in the cytoplasm. They were found free or in clusters in the cytoplasmic matrix, between the virus-induced vesicles. EV 71 particles were also occasionally observed in the form of paracrystalline arrays situated in the vesiculated areas. The immunolabel (gold beads) was found, initially, over dense cytoplasmic areas and in more advanced process at the vesiculated area and over the virus particles. Therefore the main cellular alterations observed in this infection were the shape change of the cell and the appearance of electron-dense areas (viroplasm) and smooth walled vesicles which are probably the site of the virus replication.

Cell Size↗
PubMed2004

Electron capture dissociation Fourier transform ion cyclotron resonance mass spectrometry in the electron energy range 0-50 eV.

Electron capture dissociation (ECD) of polypeptide cations was obtained with pencil and hollow electron beams for both sidekick and gas-assisted dynamic ion trapping (GADT) using Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) with an electrostatic ion transfer line. Increasing the number of trapped ions by multiple ICR trap loads using GADT improved the ECD sensitivity in comparison with sidekick ion trapping and ECD efficiency in comparison with single ion trap load by GADT. Furthermore, enhanced sensitivity made it possible to observe ECD in a wide range of electron energies (0-50 eV). The degree, rate and fragmentation characteristics of ECD FTICR-MS were investigated as functions of electron energy, electron irradiation time, electron flux and ion trapping parameters for this broad energy range. The results obtained show that the rate of ECD is higher for more energetic (>1 eV) electrons. Long electron irradiation time with energetic electrons reduces average fragment ion mass and decreases efficiency of formation of c- and z-type ions. The obtained dependencies suggest that the average fragment ion mass and the ECD efficiency are functions of the total fluence of the electron beam (electron energy multiplied by irradiation time). The measured electron energy distributions in low-energy ECD and hot ECD regimes are about 1 eV at full width half maximum in employed experimental configurations.

Electrons↗