Data for semi-permanent cationic coating for protein separations.

Protein separations and analyses are fundamental to fields of study that include biochemistry, biology, physiology, drug discovery, pharmaceuticals, as well as agricultural and food based industries. Here, we provide the data from a novel phospholipid-cetyltrimethylammonium bromide coating capable of separating cationic and anionic proteins with high efficiency. Capillary electrophoresis separations of protein standards were utilized to characterize the performance of the novel coating. Using capillary electrophoresis with UV absorbance detection a working pH range of 4-9 was identified, with reproducibility in time ≤1% relative standard deviation, and plate counts for proteins as high as 480,000 plates (lysozyme, pH 7). Further details and results from these data are available in the work reported by Crihfield et al. and can be accessed at https://doi.org/10.1016/j.chroma.2019.460397 [1].

Semi-permanent cationic coating for protein separations.

Capillary electrophoresis has been used extensively for protein separations, but interactions of proteins with the negative charge on the surface of fused silica capillary create band broadening and diminish the separation efficiency. Coatings developed to mask the negative charge of the capillary affect the electroosmotic flow. The method presented in this work addresses these concerns through the use of a two-layer coating of a semi-permanent phospholipid substrate and cetyltrimethylammonium bromide (CTAB). When used alone, phospholipid coating suppresses the electroosmotic flow but cannot be used to simultaneously separate anionic and cationic proteins. When used alone, CTAB creates a dynamic coating that facilitates the separation of cationic proteins with good efficiency, but reduces the separation efficiency of anionic proteins. The use of a hybrid phospholipid-CTAB surface coating alleviates protein adsorption, as demonstrated through a comparison of protein separations obtained with a bare fused silica capillary. The hybrid phospholipid-CTAB surface enables high efficiency separations of cationic and anionic proteins simultaneously. This work verifies the role of the hydrophobic tail of CTAB in developing a stable coating with an electroosmotic flow of 3.14 × 10-4 cm2V-1s-1 (n = 10) from the cathode to the anode at a pH of 7. The coating yields a stable electroosmotic flow even after 2 h of flushing with background electrolyte devoid of CTAB (n = 3) and six consecutive protein injections with no flush sequence between runs. The coating can be used with background electrolytes with pH values ranging from 4 to 8 while maintaining 1% RSD (n = 10) in the electroosmotic flow for each background electrolyte. Six model proteins, lysozyme, ribonuclease A, α-chymotrypsinogen A, enolase, transferrin, and α-1-antitrypsin, with pI values ranging from 4.4 to 11 were used to demonstrate the stability of the phospholipid-CTAB coating, the lack of protein interaction with the wall, and the utility of the coating for the separation of proteins of similar isoelectric points and of protein isoforms.

Taxonomic revision and multi-locus phylogeny of the North American clade of Ceratocystis.

The North American clade (NAC) of Ceratocystis includes pathogenic species that infect a wide range of woody hosts. Previous phylogenetic analyses have suggested that this clade includes cryptic species and a paraphyletic C. variospora. In this study, we used morphological data and phylogenetic analyses to characterize NAC taxa, including Ceratocystis isolates causing a serious disease of almond trees in California. Phylogenetic analyses based on six gene regions supported two new species of Ceratocystis. Ceratocystis destructans is introduced as the species causing severe damage to almond trees in California, and it has also been isolated from wounds on Populus and Quercus in Iowa. It is morphologically similar to C. tiliae, a pathogen on Tilia and the most recently characterized species in the NAC. Ceratocystis betulina collected from Betula platyphylla in Japan is also newly described and is the sister taxon to C. variospora. Our six-locus phylogenetic analyses and morphological characterization resolved several cryptic species in the NAC.

Meloidogyne aegracyperi n. sp. (Nematoda: Meloidogynidae), a root-knot nematode parasitizing yellow and purple nutsedge in New Mexico.

Meloidogyne aegracyperi n. sp. is described from roots of purple nutsedge in southern New Mexico, USA. Mature females are small (310-460 µm), pearly white, with their egg masses completely contained inside root galls. The neck is often at a 90 to 130° angle to the protruding posterior end with the perineal pattern. The distance of the dorsal esophageal gland orifice (DGO) to the base of the stylet is relatively long (4.0-6.1 µm), and the excretory pore is level with the base of the stylet. The anterior portion of the rounded lumen lining of the metacorpus contains 3 to 10 small vesicles. The perineal pattern has a rounded dorsal arch with a tail terminal area that is smooth or marked with rope-like striae. Only two males were found. The body twists 90° throughout its length. The DGO to the base of the stylet is long (3.0-3.3) µm. The cephalic framework of the second-stage juvenile is weak, and the stylet is short (10.1-11.8 µm). The DGO to the base of the stylet is long (3-5 µm). The tail is very long (64-89 µm) and the hyaline portion of the tail is very narrow, making the tail finely pointed. Eggs are typical for the genus and vary in length (85.2-99.8 µm) and width (37.1-48.1 µm), having a L/W ratio of (2.1-2.6). Maximum likelihood phylogenetic analyses of the different molecular loci (partial 18S rRNA, D2-D3 of 28S rRNA, internal transcribed spacer (ITS) rRNA, cytochrome oxidase subunit II (COII)-16S rRNA of mitochondrial DNA gene fragments and partial Hsp90 gene) placed this nematode on an independent branch in between M. graminicola and M. naasi and a cluster of species containing M. chitwoodi. M. fallax, and M. minor. Greenhouse tests showed that yellow and purple nutsedge were the best hosts, but perennial ryegrass, wheat, bentgrass, and barley were also hosts.

Obesity is associated with impaired immune response to influenza vaccination in humans.

BACKGROUND: Obesity is an independent risk factor for morbidity and mortality from pandemic influenza H1N1. Influenza is a significant public health threat, killing an estimated 250,000-500,000 people worldwide each year. More than one in ten of the world's adult population is obese and more than two-thirds of the US adult population is overweight or obese. No studies have compared humoral or cellular immune responses to influenza vaccination in healthy weight, overweight and obese populations despite clear public health importance. OBJECTIVE: The study employed a convenience sample to determine the antibody response to the 2009-2010 inactivated trivalent influenza vaccine (TIV) in healthy weight, overweight and obese participants at 1 and 12 months post vaccination. In addition, activation of CD8⁺ T cells and expression of interferon-γ and granzyme B were measured in influenza-stimulated peripheral blood mononuclear cell (PBMC) cultures. RESULTS: Body mass index (BMI) correlated positively with higher initial fold increase in IgG antibodies detected by enzyme-linked immunosorbent assay to TIV, confirmed by HAI antibody in a subset study. However, 12 months post vaccination, higher BMI was associated with a greater decline in influenza antibody titers. PBMCs challenged ex vivo with vaccine strain virus, demonstrated that obese individuals had decreased CD8⁺ T-cell activation and decreased expression of functional proteins compared with healthy weight individuals. CONCLUSION: These results suggest obesity may impair the ability to mount a protective immune response to influenza virus.

Microscale exoglycosidase processing and lectin capture of glycans with phospholipid assisted capillary electrophoresis separations.

Capillary electrophoresis separations of glycans labeled with 1-aminopyrene-3,6,8-trisulfonic acid were achieved with separation efficiencies ranging from 480,000 to 640,000 theoretical plates in a 60.2 cm, 25 µm inner diameter fused silica capillary. Under these separation conditions, the coefficient of variation in peak area is 10%, and if labeling efficiency is estimated at 100%, the limit of detection is 15 fM. The capillary electrophoresis method incorporated phospholipid additives to enhance the separation of glycans with slight differences in hydrodynamic volume. In addition, the phospholipid additives supported the integration of the lectin concanavalin A as well as the enzymes α1-2,3 mannosidase or ß1-4 galactosidase to provide structural and compositional information about the glycans subject to separation. The use of in-capillary cleavage of terminal glycan residues with exoglycosidases offers a number of advantages over benchtop enzymatic sequencing, including reduced consumption of analyte, as well as enzyme. These methods were used to evaluate glycans derived from the glycoproteins α1-acid glycoprotein, fetuin, and ribonuclease B, as well as from glycoproteins collected from MCF7 cells.

An integrated electrochemical capillary liquid chromatography-dual microelectrode system for bromine based reaction detection.

Amperometric reaction detection of model peptides containing either a thiol or disulfide was successfully achieved with a novel coupled capillary liquid chromatography-dual microelectrode system. Detection is facilitated via dual platinum wire electrodes fabricated directly on a liquid chromatography capillary column. The detection strategy is based on the electrogeneration of bromine at the upstream working electrode followed by reduction of the bromine to bromide at the downstream working electrode. When bromine reacts with analyte eluting from the capillary column, the amount of bromine that reaches the downstream electrode decreases. As a result, the current at the downstream electrode will decrease in proportion to the amount of analyte eluting from the capillary column. Glutathione, glutathione disulfide, alpha-TGF and oxytocin were used to characterize the system and to determine system parameters. The feasibility of the determination of disulfide containing peptides is demonstrated with glutathione disulfide, alpha-TGF and oxytocin.

Classification models of child molesters utilizing the Abel Assessment for sexual interest.

OBJECTIVE: The aims of this study are to demonstrate 1) the criterion validity of the Abel Assessment for sexual interest (AASI) based on its ability to discriminate between non child molesters and admitting child molesters, and 2) its resistance to falsification based on its ability to discriminate between liar-denier child molesters and non child molesters. METHOD: A group of 747 participants matched by age, race, and income was used to develop three logistic regression equations. The models compare a group of non child molesting patients under evaluation for other paraphilias to three groups: 1) a group of admitting molesters of girls under 14 years of age, 2) a group of admitting molesters of boys under 14 years of age, and 3) a group believed to be concealing or denying having molested. RESULTS: Both of the equations designed to discriminate between admitting child molesters and non child molesters were statistically significant. The equation contrasting child molesters attempting to conceal or deny their behavior and non child molesting patients was also statistically significant. CONCLUSIONS: The models classifying admitting child molesters versus non child molesters demonstrate criterion validity, while the third model provides evidence of the AASI's resistance to falsification and its utility as a tool in the detection of child molesters who deny the behavior. Results of the equations are reported and suggestions for their use are discussed.

Obtaining event status at the close of the Treatment of Mild Hypertension Study: methods and implication for other trials.

The Treatment of Mild Hypertension Trial was a randomized, double-blind clinical trial conducted from 1986 to 1992 comparing the efficacy of six antihypertensive treatment regimens in 902 participants with stage I hypertension. To satisfy a secondary objective of the study, follow-up information on mortality and cardiovascular morbidity was collected. For this objective the aim was to ascertain the vital and cardiovascular event status as of the last day of the trial. This was accomplished by inviting each participant to attend a closeout visit shortly after the closeout date. In addition to serving as verification of vital status, this visit allowed data collection on nonfatal events that occurred between the last clinic visit and the closeout date. During this visit the patient was unblinded to study medication and given a medical summary of their participation during the trial, as well as a bottle of open-label medication. The advantages of a closeout visit are discussed along with a call for studies to provide clearer definitions of lost to follow-up and censoring times used in life-table analyses, especially when the primary event includes both fatal and nonfatal events. Control Clin Trials 2001;22:56-61

Psychometric analysis of the Sexual Interest Cardsort Questionnaire.

Reliability, measured by Cronbach's coefficient alpha, and concurrent validity, measured by Pearson's r and polychoric correlation coefficients, were evaluated in this study. A sample of 371 sexual offenders referred to the Behavioral Medicine Institute of Atlanta for evaluation of sexual interests and behaviors by the courts were assessed using the Sexual Interest Cardsort Questionnaire (SI), a self-report measure of deviant and nondeviant sexual interest, as well as indicator variables obtained from classifications assigned by clinicians as a result of 2 hour-long, semistructured clinical interviews. Internal consistency of 75 items from the SI ranged from 0.71 to 0.96, across 15 categories of sexual interest and behavior. Additionally, the SI was shortened utilizing Cronbach's alphas to maintain a high level of internal consistency. The resulting questionnaire, the shortened SI (SIS), had 45 items and 15 categories. Cronbach's alpha ranged from 0.78 to 0.97. Utilizing Pearson's r and polychoric correlation coefficients, significant correlations were found for the 11 sexually deviant categories of the SI and indicator variables, and the 10 sexually deviant categories of the SIS and indicator variables. The SI and SIS showed a high level of reliability and concurrent validity. Clinical and research issues pertaining to the clinical assessment of male sexual offenders utilizing self-report and clinical interview data, both obtained as the result of comprehensive evaluations, are discussed.

Postcolumn reaction detection with dual-electrode capillary electrophoresis-electrochemistry and electrogenerated bromine.

This is the first report of postcolumn amperometric reaction detection for capillary electrophoresis and dual-electrode detection. Bromide present in the run buffer is oxidized to bromine at the first electrode and subsequently detected at a second electrode downstream. Analytes that react with bromine cause a decrease in signal at the downstream electrode that is proportional to analyte concentration. Bromine is known to react with a variety of compounds, including thiols, thioethers, disulfides, amines, and unsaturated organic compounds. In this paper, the development of a new wire--wire on-capillary dual electrode that is well suited to bromine-based post-column reaction detection is described. System performance was evaluated using glutathione, cysteine, and methionine as test analytes. The final optimized system could be operated continuously for 24 h and was stable for day-to-day use for at least two weeks. The response for cysteine was linear from 0.5 to 20 microM with a limit of detection of approximately 80 nM.

Capillary electrophoresis in pharmaceutical analysis.

Capillary electrophoresis (CE) is a separation technique particularly suited to the analysis of pharmaceutical compounds. This review offers a detailed discussion of the four common modes of detection coupled to CE-UV absorption, fluorescence, electrochemical, and mass spectrometry-and gives examples of the use of these methods in pharmaceutical analyses. Sample preparation and pretreatment techniques used for CE separations are described, as well as methods of preconcentration including hydrophobic retention, affinity concentration, sample stacking, and isotachophoresis. The use of affinity CE, chiral CE, and capillary gel electrophoresis for analysis of pharmaceuticals is covered in detail, and recent advances in capillary electrochromatography and CE on a chip are also discussed.

Separation of nanoliter samples of biological amines by a comprehensive two-dimensional microcolumn liquid chromatography system.

A two-dimensional liquid chromatography system based on the combination of a charge separation mechanism, provided by anion exchange chromatography, with a hydrophobic separation mechanism, provided by reversed phase chromatography, is presented. A 90 cm long anion exchange microcolumn is coupled to a 3 cm long reversed phase microcolumn. Both microcolumns are interfaced by two electronically controlled valves. Effluent from the anion exchange microcolumn is collected in a sample loop and then concentrated onto the head of the reversed phase microcolumn. Analyte peaks consisting of tagged amines elute from the second column and are detected by laser-induced fluorescence. The entire operation is controlled by a personal computer. The resolving power of this system is demonstrated with a two-dimensional chromatogram of the peptides obtained from a tryptic digest of porcine thyroglobulin. The sensitivity of the system is demonstrated with a two-dimensional chromatogram of the contents of a single bovine chromaffin cell.

Postnatal depression: a Christchurch study.

AIMS: The aims of the study were to enquire into the incidence of postnatal depression (PND) and factors associated with it, in a New Zealand population. METHODS: A wide ranging postal questionnaire, which included the Edinburgh postnatal depression scale (EPDS), was administered to a sample of 1330 women 6-9 months after the birth of their babies at Christchurch Women's Hospital, in 1991/2. RESULTS: Measured on the EPDS, 80% of the women were not depressed, 7% were at a threshold level of depression and 13% were more severely depressed. Only 6% recognised their symptoms as depression. The factors having the greatest association with high depression scores were: depressive symptoms before and during the pregnancy; serious deterioration in the partner relationship after the birth; decreased energy, confidence and happiness levels after the pregnancy; moderate or severe premenstrual tension prepregnancy; frequent nausea in later pregnancy; low education and low income. CONCLUSIONS: Postnatal depression appears to be more widespread than is generally realised. The findings suggest that, at the time of postnatal checks on the baby, greater use could be made of the EPDS to increase confidence in a diagnosis of depression. Involvement of male partners in the treatment process is highly desirable.

A dc Method for the Absolute Determination of Conductivities of the Primary Standard KCl Solutions from 0 °C to 50 °C.

A new method for the absolute determination of electrolytic conductivity based on direct current and Potentiometric measurements is described. The unique design of the cell uses a removable center section whose length and cross-sectional area are accurately known. Two pairs of matched Ag, AgCl electrodes are used in a four terminal mode of resistance measurement. Measurements of the electrolytic conductivity of primary standard potassium chloride solution using [his novel dc conductance cell are compared with the currently adopted IUPAC and OIML recommendations. In addition, measurements have heen made of the electrolytic conductivity of a solution of potassium chloride having a molality of 1 mol/kg (mole KCl per kilogram H2O). The values so obtained over the temperature range of 0 °C to 50 °C are recommended as the new primary standards for electrolytic conductivity.

Waterfowl mortality caused by aflatoxicosis in Texas.

Waterfowl mortality caused by aflatoxicosis occurred in two separate areas in Texas during the 1977-78 wintering season. The first outbreak occurred in snow geese (Anser caerulescens) on the Gulf Coast prairies, followed by an outbreak in mallards (Anas platyrhynchos) in the north-central portion of the state. Aflatoxin B1 levels in geese were 500 ng/g (dry weight). Aflatoxin B1 levels in the second mortality were 10-250 ng/g (dry weight). The exact source of the toxin was not demonstrated in the first outbreak, but in the second outbreak was traced to waste peanuts, which constituted a major portion of the diet of wintering waterfowl in north-central Texas. Aflatoxin B1 levels in the field peanuts collected in the general areas were 110 ng/g.