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At least 1,315 records · Page 73Linked to original sources
PubMed1997

Modified APC-resistance test: variable ratios with respect to source of factor V-deficient plasma.

A single point mutation of the factor V (FV) gene, leading to the substitution Arg506Gln in the FV molecule (FV-Leiden) and hence resistance to its breakdown by activated protein C (APC), is the most prevalent risk factor for venous thrombosis in the Caucasians. A ratio determined by activated partial thromboplastin time (APTT) of test plasma in the presence or absence of exogenous APC (the APC ratio), is the method widely used to screen individuals with this risk factor for thrombosis. Because of functional defects of vitamin K-dependent clotting factors in patients on oral anticoagulant therapy, this method cannot be applied to those patients without modification. One modification is to mix test plasma (1:5 or 1:10) with FV-deficient plasma so that 80-90% of functioning vitamin K-dependent factors are supplied by the FV-deficient plasma. Even with 10-20% of FV in the mixture, APC-resistance still can be demonstrated. In this report, we present our results of the modified APC-sensitivity assay using FV-deficient plasma from different commercial sources. APC ratios determined by the original method in which test plasma is not mixed with FV-deficient plasma can be significantly different from those determined by the modified method in which test plasma is diluted 1:5 with FV-deficient plasma. This difference between methods was observed not only in normal individuals, but also in FV-Leiden positive individuals, and in patients on warfarin therapy. Further, APC ratios varied significantly depending on the commercial source of the FV-deficient plasma. The modified method is apparently suitable to identify APC-resistance in patients on warfarin therapy, as well as in individuals not receiving anticoagulant treatment. However, one must be aware that APC-resistance ratios obtained with the modified method are likely to be different from those established with the original method, and the source of FV-deficient plasma can be a factor influencing the ratios in the former cases.

Anticoagulants↗
PubMed1996

Modified heparin-bonded catheter for cannulation of the coronary sinus from the femoral vein.

Serial sampling from the coronary sinus is an attractive technique for drawing blood samples to be used in the characterization of procoagulant activity during coronary interventions. We have developed a modified Simmons catheter for rapid cannulation of the coronary sinus from the femoral approach. The catheter design incorporates heparin bonding and distal side-holes to minimize blood sampling artifacts. Coronary sinus cannulation was performed via the femoral approach in 186 patients by use of a multipurpose catheter (n = 8), an unmodified Simmons I or II catheter (n = 64), or the modified Simmons catheter (n = 114). The coronary sinus was cannulated successfully with the modified Simmons catheter in 97% of patients; the success rate with the unmodified Simmons II catheter was 87% (P = 0.02). The modified Simmons catheter represents an improved technique for cannulation of the coronary sinus from the femoral vein.

Adult↗
PubMed1996

Glow-discharge-modified activated carbon cloths as skin substitutes.

In this study, activated carbon cloths (ACCs) were modified in a glow-discharge apparatus by using 2-hydroxyethyl-methacrylate (HEMA), dimethyl-aminoethyl methacrylate (DMAEMA), and hydroxymethyldisiloxane (HMDS) plasmas. Scanning electron micrographs exhibited the formation of the polymeric films on the ACCs after glow-discharge modification. The adsorption experiments were performed using model solutes, i.e., creatinine, vitamin B-12, and trypsin. No significant change was observed in the adsorption capacities of the ACC/PHEMA and ACC/PDMAEMA composites compared with the original ACC adsorption capacity. However, there was a pronounced decrease in the case of the ACC/PHMDS. Oxygen permeabilities of both the original and modified ACCs were measured in a flow-through gas chamber. The oxygen permeabilities of the original and modified forms were 817 and 3.5-5 cm3/s.cm2.cm Hg x 10(-7), respectively. Water-vapor permeabilities of the composites were obtained by using a standard water-cup assembly. Permeabilities varying between 921 and 10,300 g/m2.day were found. Microorganism impermeabilities of the ACCs were demonstrated by using model PS monosize polymeric particles with a diameter of 0.2 micron. The particle permeability of the original ACC was 65.6 mg/m2.day. Almost no particle permission was observed through the modified ACCs.

Adsorption↗
PubMed1999

Acid-base complex reactions in resin-modified and conventional glass ionomer cements.

Resin-modified glass ionomer cements are a recent development in which the desirable properties of glass ionomer cements and resin-composites are combined. The presence of resin may, however, retard the acid-base reaction of the ionomer component. This has led to a debate regarding the classification of resin modified materials as true glass ionomer cements, and the actual duration of the acid-base reaction after initial setting via light polymerization has taken place. To investigate this issue, a novel method employing FT-IR spectrophotometry was used to monitor the acid-base complexation reaction in a resin-modified glass ionomer cement (Fuji II LC) and a conventional, chemically cured cement (Fuji II Cap). This method involved subtraction of the background "resin" spectrum from the Fuji II LC spectra and subsequent application of a baseline to obtain the plot of absorbance area ratio in the range of 1685 to 1510 cm(-1) (complexed carboxyls) to that in the range of 1750 to 1685 cm(-1) (free carboxyls). This study demonstrates evidence of a delayed acid-base reaction for the resin-modified cement, which levels off after 168 hours of cement mixing. In contrast, the complexation reaction of the conventional glass ionomer cement was essentially complete after 24 hours.

Glass↗
PubMed2000

A modified nasopharyngeal tube to relieve high upper airway obstruction.

Infants with high upper airway obstruction (UAO) are managed with a variety of techniques to relieve their UAO. Among these techniques, the least invasive and safest is the nasopharyngeal tube (NPT). However, the traditional NPT is not always satisfactory, and tracheostomies need to be done. We recently described a modified NPT technique that, in contrast to the traditional tube, does not add airway dead space and resistance, is easy to use, is well-tolerated, has proven highly successful, and allows the simultaneous use of oxygen nasal prongs. This modified NPT has many advantages over the traditional NPT as a temporary management of high UAO that resolves with growth of the infant. This report highlights the respiratory care of 10 infants with high UAO (Pierre Robin syndrome, Down syndrome, Goldenhar syndrome, isolated microngathia, and idiopathic hypotonia) who were managed with a modified NPT. The modified NPT described potentially reduces the need for surgical intervention to relieve high UAO in infants.

Airway Obstruction↗
PubMed1998

Production of genetically modified lysozymes having extreme heat stability and antimicrobial activity against gram negative bacteria in yeast and in plant.

Hen egg white lysozyme was genetically modified to have extreme heat stability and strong antimicrobial activity against Gram negative bacteria and the modified lysozymes were secreted in yeast and tobacco. Complementary DNA encoding lysozyme was subjected to site-directed mutagenesis to have the Asn-X-Thr(Ser) sequence that is the signal for asparagine-linked glycosylation at the positions 49. The glycosyl lysozyme enhanced heat stability was expressed in the yeast carrying the modified lysozyme cDNA. The expression amount of glycosyl lysozyme was about 10 mg/l of yeast culture medium. Using the same yeast expression system, the lysozyme enhanced antimicrobial action by inserting hydrophobic penta-peptide at the C-terminus were secreted in a small amount (less than 100 micrograms/l in the yeast culture medium). These cDNA constructs of modified lysozymes were engineered into tabacco through Agrobacterium-mediated transformation in order to construct antimicrobial plant. The expression of lysozymes was confirmed by the reverse transcriptional PCR, SDS-PAGE analysis and lytic activity of transformants of tobacco. The transformant having the highest lytic activity expressed about 40 micrograms of lysozyme per g of leaf tissue.

Anti-Bacterial Agents↗
PubMed1986

Is modified radical mastectomy really equivalent to radical mastectomy in treatment of carcinoma of the breast?

Evaluation of any therapeutic modality for carcinoma of the breast requires prolonged follow-up. Between 1965 and 1968, 870 women with primary unilateral breast cancer were treated surgically at our institution. To achieve a study group that could be meaningfully analyzed, the authors identified and excluded all patients with pre-existing life-threatening disease, incomplete (palliative) operation, multiple lesions, or fixed lesions. There remained 530 patients (322 treated with radical and 208 treated with modified radical mastectomy). No overall statistical difference in 5-year survival to death from breast cancer was noted between the radical (82.1%) and the modified radical (87.0%) mastectomy group. Recently, the authors updated that series, now with 10 to 16 years of follow-up (minimum, 10 years). Two hundred eighty-one patients were alive at 10 years. The 10-year survival from breast cancer after radical (74.5%) and modified radical (74.2%) mastectomy was almost identical. The 10-year node-negative survival was 86.0% and the node-positive survival was 56.3% (P = 0.0001). Results were compared according to age, tumor site, absolute number of involved nodes, histologic features, grade, size, and adjuvant treatment. A numeric combination of these variables was constructed and used as a prognostic score. These results suggest that, with equal long-term efficacy in the treatment of breast cancer, the cosmetically and functionally superior modified radical mastectomy should remain the procedure of choice in the surgical treatment of carcinoma of the breast.

Adult↗
PubMed1984

The utility of the modified WAIS in a clinical setting.

Utilized time-saving WAIS scoring procedures in two ex post facto investigations following approaches outlined by Vincent (1979) and Himelstein (1983). These procedures direct an examiner to use an S's results on the Information subtest as a criterion for determining the point of initial questioning on other scales. Study 1 followed exactly the procedure presented by Himelstein (1983), with an obtained correlation between modified WAIS and WAIS Verbal IQ of .993. In spite of the high correlation, however, significant distortion in results occurred for some Ss, Study 2 integrated and modified slightly the methodologies of Vincent (1979) and Himelstein (1983). Obtained correlations between modified WAIS and WAIS Verbal, Performance, and Full Scale IQs (N = 40) were each .998. The percentage of IQs that differed by more than 2 points between Modified WAIS and WAIS for Verbal, Performance, and Full Scale scores was 0%, 5%, and 2.5%, respectively. In addition, the procedure maintained the integrity of subscale configurations.

Adolescent↗
PubMed2000

A modified silver staining protocol for visualization of proteins compatible with matrix-assisted laser desorption/ionization and electrospray ionization-mass spectrometry.

The growing availability of genomic sequence information, together with improvements in analytical methodology, have enabled high throughput, high sensitivity protein identification. Silver staining remains the most sensitive method for visualization of proteins separated by two-dimensional gel electrophoresis (2-D PAGE). Several silver staining protocols have been developed which offer improved compatibility with subsequent mass spectrometric analysis. We describe a modified silver staining method that is available as a commercial kit (Silver Stain PlusOne; Amersham Pharmacia Biotech, Amersham, UK). The 2-D patterns abtained with this modified protocol are comparable to those from other silver staining methods. Omitting the sensitizing reagent allows higher loading without saturation, which facilitates protein identification and quantitation. We show that tryptic digests of proteins visualized by the modified stain afford excellent mass spectra by both matrix-assisted laser desorption/ionization and tandem electrospray ionization. We conclude that the modified silver staining protocol is highly compatible with subsequent mass spectrometric analysis.

Animals↗
PubMed2001

Estimation of the dissociation constants for functional groups on modified and unmodified silica gel supports from the relationship between electroosmotic flow velocity and pH.

Electroosmotic volume flow was directly observed in a simple instrument consisting of 1 cm long a home-made support, packed between two polyethylene frits in the polypropylene tube. Equations relating electroosmotic flow (EOF) velocity and pH for two functional groups on the surface of the solid materials were developed. With these equations, we can estimate the dissociation constants of two different kinds of functional groups on modified silica gel materials simultaneously. The dissociation constants of silanol groups, benzene sulfonic acid groups, and alkyl quaternary ammonium groups on the modified and unmodified silica gel supports were estimated. The estimated pK values of the silanol groups on the silica gel and modified silica gel surfaces are between 4.0 and 4.3. The estimated pK values of the benzene sulfonic acid groups and alkyl quaternary ammonium groups on the surface of the modified silica gel are 2.6 and 8.6, respectively.

Hydrogen-Ion Concentration↗
PubMed2002

Identification of drug-binding sites on human serum albumin using affinity capillary electrophoresis and chemically modified proteins as buffer additives.

A technique based on affinity capillary electrophoresis (ACE) and chemically modified proteins was used to screen the binding sites of various drugs on human serum albumin (HSA). This involved using HSA as a buffer additive, following the site-selective modification of this protein at two residues (tryptophan 214 or tyrosine 411) located in its major binding regions. The migration times of four compounds (warfarin, ibuprofen, suprofen and flurbiprofen) were measured in the presence of normal or modified HSA. These times were then compared and the mobility shifts observed with the modified proteins were used to identify the binding regions of each injected solute on HSA. Items considered in optimizing this assay included the concentration of protein placed into the running buffer, the reagents used to modify HSA, and the use of dextran as a secondary additive to adjust protein mobility. The results of this method showed good agreement with those of previous reports. The advantages and disadvantages of this approach are examined, as well as its possible extension to other solutes.

2-Hydroxy-5-nitrobenzyl Bromide↗
PubMed2026

Molecular Mobility of N-Acetylgalactosamine-Modified Cyclodextrins on a Polyrotaxane for Highly Efficient Liver Targeting of Antibody Chimeras and Genome-Editing Ribonucleoproteins.

Triantennary N-acetylgalactosamine (triGalNAc), which interacts strongly with the trimeric structure of asialoglycoprotein receptors (ASGPRs), is a validated platform for liver targeting. However, the intricate design and synthesis of its linkers impose high production costs and significant technical challenges. In this study, we report an alternative strategy for targeting ASGPR using monovalent GalNAc (monoGalNAc) conjugated to the cyclic molecules of polyrotaxane, which can rotate and translocate along the axial polymer chain. The intracellular uptake efficacy of monoGalNAc-modified polyrotaxane is comparable to that of triGalNAc-modified polyrotaxane and significantly higher than that of triGalNAc- or monoGalNAc-modified immobile control polymers. These results suggest that the inherent mobility of polyrotaxanes allows monoGalNAc moieties to cluster in a trivalent-like manner, thereby enhancing multivalent interactions with multiple ASGPR oligomers. The successful application of monoGalNAc-modified polyrotaxane to lysosome-targeting antibody chimeras and genome-editing nanoparticles demonstrates that this facile technology is a highly promising alternative to conventional triGalNAc.

Rotaxanes↗
PubMed1983

Studies on the sequestration of chemically and enzymatically modified erythrocytes.

We have developed an experimental animal model to establish the patterns of sequestration of untreated, as well as chemically and enzymatically modified red blood cells (RBC). The intraperitoneal route of transfusion provides a useful way of transferring large numbers of untreated RBC into small animals and assuring their introduction into the circulation within 24 hr. Moreover, this route "filters" some types of modified erythrocytes, eg, glutaraldehyde treated RBC. From the pattern of sequestration, the RBC in the peritoneal cavity then pass through the liver where other types of modified RBC are sequestered, eg, after trypsin, pronase, protease, or sialidase treatment. Some modified RBC show a preference for the spleen as the site of sequestration, eg, galactose oxidase or N-ethylmaleimide treated RBC. These appear in the spleen despite intraperitoneal transfusion. Relevant to this study is the observation that in the rat old RBC are sequestered both by liver and spleen, while asialoerythrocytes are sequestered by liver only. A possible reason for this difference is discussed in the text.

Animals↗
PubMed1988

A modified method of DNA extraction from peripheral blood and bone marrow specimens.

We have outlined a modified method of DNA extraction from blood and bone marrow that can be used for gene rearrangement studies of leukemia and lymphoma and can consistently give excellent yields of DNA from as little as 1 x 10(6) cells. The method is based on the use of small sample vessels, a marked reduction in solution volume and minimal handling of the specimen. By using known cell numbers from eight leukemic bone marrow samples and four leukemic cell lines, the modified extraction procedure consistently had better yields of DNA, compared with a standard procedure. These findings were confirmed by further comparisons of DNA yields from 14 hypocellular clinical specimens that had been divided into two equal parts for side-by-side DNA extraction with both methods. The quality of the DNA from the modified extraction method was similar to standard methods on subsequent Southern blot analysis and hybridization with immunoglobulin and/or T-cell receptor gene probes. This modified DNA extraction method is a procedure that complements standard extraction methods and expands the number and type of specimens that can be evaluated by the clinical molecular genetics laboratory.

Blood Cell Count↗
PubMed2004

Invasive cleavage reactions on DNA-modified diamond surfaces.

Recently developed DNA-modified diamond surfaces exhibit excellent chemical stability to high-temperature incubations in biological buffers. The stability of these surfaces is substantially greater than that of gold or silicon surfaces, using similar surface attachment chemistry. The DNA molecules attached to the diamond surfaces are accessible to enzymes and can be modified in surface enzymatic reactions. An important application of these surfaces is for surface invasive cleavage reactions, in which target DNA strands added to the solution may result in specific cleavage of surface-bound probe oligonucleotides, permitting analysis of single nucleotide polymorphisms (SNPs). Our previous work demonstrated the feasibility of performing such cleavage reactions on planar gold surfaces using PCR-amplified human genomic DNA as target. The sensitivity of detection in this earlier work was substantially limited by a lack of stability of the gold surface employed. In the present work, detection sensitivity is improved by a factor of approximately 100 (100 amole of DNA target compared with 10 fmole in the earlier work) by replacing the DNA-modified gold surface with a more stable DNA-modified diamond surface.

Apolipoproteins E↗
PubMed2002

Miniaturized amperometric flow immunoassay system using a glass fiber membrane modified with anion.

This paper describes a miniaturized amperometric flow immunoassay system using a glass fiber membrane modified with anion. The glass fiber membrane was functionally modified with gamma-glycidoxypropyltrimethoxysilane and sodium thiosulfate and was used for separation of protein. Anti-human chorionic gonadotrophin (HCG) immunoglobulin G (IgG) antibody conjugated with ferrocenemonocarboxylic acid (Fc), namely, Fc-conjugated IgG (Fc-IgG), was used as a novel analytical reagent. HCG and Fc-IgG complexes were separated from free Fc-IgG based on differences in isoelectric point (pI) using the glass fiber membrane modified with a thiosulfonyl acid functional group. The assay yields a linear relationship between current and HCG concentration in the range of 0-2000 mIU/mL. This simple technique enables the assay of HCG within 2 min. The modified glass fiber membrane was regenerated by occasional elution with malonate buffer (pH 6.0) containing 0.5 M NaCl, to remove free Fc-IgG. Free Fc-IgG recovered in this manner could be reused up to eight times without significant decreases in sensitivity. This miniaturized amperometric flow immunoassay requires only minute quantities of serum and generates highly reproducible results.

Animals↗
PubMed2006

Phytol-modified heme in mesoporous silica: conjugates as models of hemoproteins.

A ferriprotoporphyrin, hemin (Fe(3+)), modified with 3,7,11,15-tetramethyl-2-hexadecen-1-ol, phytol, was adsorbed in nano-spaces of about 4 nm in diameter in mesoporous silica (FSM; folded-sheet mesoporous material) forming a phytol-modified hemin (Fe(3+))-FSM nano-conjugate. The properties and the structure of the conjugate were studied by UV-visible light absorption, IR absorption spectroscopy, and a nitrogen adsorption isotherm. Although the hemin without phytol could not be adsorbed to the mesoporous silica, modification with phytol imparted preferential adsorption properties. The conjugate was not only stable but also had a peroxidase-like activity in a 0.1% hydrogen peroxide solution, while free hemin in the solution was easily destroyed. The hemin (Fe(3+)) in the FSM was reduced to heme (Fe(2+)) by hydrazine. The phytol-modified heme (Fe(2+))-FSM conjugate formed an O(2)-heme complex with a superoxide type structure, resembling oxyhemoglobin or oxymyoglobin, which has not been previously observed for free heme in solution. The addition of carbon monoxide or nitrogen monoxide to the phytol-modified heme (Fe(2+))-FSM conjugate caused the formation of CO- or NO-heme complex in the nano-spaces of the FSM. These properties are attributed not only to the Fe-complex but also to the cooperative functions of the heme with mesoporous silica, resembling properties of a natural heme-protein conjugate; hemoglobin or peroxidase. These results are an elegant example of biomimetic nano-technology.

Adsorption↗
PubMed2004

Quantitation of normal and formaldehyde-modified deoxynucleosides by high-performance liquid chromatography/UV detection.

A sensitive and selective method was developed for the first time to quantify simultaneously the normal and formaldehyde (FA)-modified bases in human placental DNA treated with 100 ppm FA for 20 h at 37 degrees Celsius. Digestion of DNA to deoxynucleosides with DNase I, phosphodiesterase and alkaline phosphatase occurred in that order with centrifugation steps. The normal and FA-modified deoxynucleosides were then resolved from one another and reagent blank interferences to produce selective separation through high performance liquid chromatography-ultraviolet detection at 254 nm. A C(18) reversed-phase column facilitated the resolution using 5 mm ammonium acetate and a gradient of 0-6% methanol at fl ow rates of 0.3-1.4 mL/min before column cleaning. The lower quantifiable limits for deoxyadenosine, deoxyguanosine, deoxycytidine, thymidine, N(6)-hydroxymethyldeoxyadenosine (N(6)-dA), N(2)-hydroxymethyldeoxyguanosine (N(2)-dG) and N(4)-hydroxymethyldeoxycytidine (N(4)-dC) were 11, 7.6, 12, 15, 10, 10 and 22 pmol, respectively. The abundance order of the modified deoxynucleosides was N(6)-dA > N(2)-dG > N(4)-dC. dT did not form hydroxymethyl derivatives. The respective concentrations were about 6.0, 10.0 and 23 pmol of modified deoxynucleosides in 80 micro g of human placental DNA after treatment with 100 micro g/mL of formalin for 20 h at 37 degrees Celsius. The stabilities of N(6)-dA and N(2)-dG were much better at -20 degrees Celsius than at 25 degrees Celsius, where the respective halftimes were about 50.1 and 21.0 h.

Chromatography, High Pressure Liquid↗