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At least 1,603 records · Page 89Linked to original sources
PubMed2000

Urinary morbidity with a modified peripheral loading technique of transperineal (125)i prostate implantation.

PURPOSE: Analysis of urinary morbidity within the first 12 months following a modified peripheral loading technique for permanent transperineal transrectal ultrasound (TRUS) guided (125)I prostate implantation and comparison of urinary morbidity with various clinical and implant parameters. MATERIALS AND METHODS: Between October 1, 1996, and March 11, 1998, 87 patients with favorable, early stage prostate cancer were treated with permanent transperineal TRUS guided (125)I prostate implantation. A peripheral loading technique was utilized for source placement with 75-80% source distribution in the periphery and 20-25% source distribution centrally. A mean total activity of 38 mCi of (125)I was implanted (range, 19-66 mCi). The mean source activity was 0.43 mCi/source (range, 0.26-0.61 mCi/source) and the mean number of sources implanted was 88 (range, 56-134). The minimum prescribed dose to the prostate was 145 Gy. The median D(90), V(100), and V(150) were 152 Gy (range, 104-211 Gy), 92% (range, 71-99%), and 61% (range, 11-89%), respectively. The median follow-up time was 19 months (range, 12-29 months). Urinary morbidity was scored at 3 weeks and then at 3-month intervals for the first 2 years using a modified Radiation Therapy Oncology Group (RTOG) grading system (scale 0-5). RESULTS: Most patients developed at least minor urinary symptoms with frequency or nocturia being the most common. Overall, 79% (69/87) of patients experienced urinary morbidity with 21% (18/87) reporting no symptoms. The incidence of overall Grade 1 urinary morbidity was 37% (32/87); Grade 2 morbidity was 37% (32/87); and Grade 3 morbidity was 6% (5/87). There was no Grade 4 or 5 morbidity. The incidence of Grade 0 frequency/nocturia was 36% (31/87); Grade 1 was 33% (29/87); Grade 2 was 30% (26/87); and Grade 3 was 1% (1/87). Grade 0 dysuria was seen in 56% (49/87) of patients; 32% (28/87) had Grade 1; 10% (9/87) Grade 2; and 1% (1/87) Grade 3 dysuria. Most urinary symptoms started a few weeks after implantation and began to subside by 6 months. At 12 months, 22% (19/87) of patients had persistent urinary symptoms (78% Grade 0, 15% Grade 1, 3% Grade 2, and 3% Grade 3). The mean urethral point dose was 174 Gy (range, 99-315 Gy). The mean number of sources implanted correlated significantly with the likelihood of developing acute urinary morbidity (p = 0.03). The total activity implanted also correlated with the morbidity outcome dysuria (p = 0.01) with a threshold seen at 37 mCi. Urethral point dose, source activity, intraoperative TRUS prostate volume, D(90), V(100), V(150), patient age, pretreatment PSA, Gleason score, and T stage did not correlate with morbidity. CONCLUSIONS: Permanent transperineal TRUS guided (125)I prostate implantation using a modified peripheral loading technique is associated with mild urinary morbidity that resolves in 78% of patients by 12 months. Grade 3 urinary morbidity was encountered in only 6% (5/87) of patients. Urinary morbidity may be related to the total number of sources implanted and/or the total activity implanted. Overall urinary morbidity was not correlated with urethral point dose, source activity, intraoperative TRUS prostate volume, D(90), V(100), V(150), patient age, pretreatment PSA, Gleason score, and T stage. The low incidence of urinary morbidity may be a consequence of our modified peripheral loading technique and/or the selection of patients with good-to-excellent preimplant urological parameters. Longer follow-up is necessary to assess biochemical control rates and long-term morbidity.

Aged↗
PubMed2003

Direct comparison of microarray gene expression profiles between non-amplification and a modified cDNA amplification procedure applicable for needle biopsy tissues.

Global gene expression profiling by cDNA microarray analysis has been used to discover the biomarkers for early diagnosis of various cancers, subclassing cancer type, and prediction of patient's treatment outcome. The information provided by gene expression profiling may contribute to the design of molecular mechanism-based strategies for cancer prevention and/or treatment. However, the standard procedure for cDNA microarray analysis requires 5mug of good quality total RNA as starting material for each target preparation reaction. Thus, there is a limit for needle biopsy samples, laser capture microdissected tissues, or flow-sorted cells to successfully utilize the microarray technology. In order to profile the gene expression of needle biopsy tissue, we have modified the standard protocol by carrying out cDNA amplification after cDNA synthesis. We compared percentage present calls, absent calls, reproducibility, and concordance in needle biopsy samples processed by standard microarray protocol (cDNA non-amplification method) and our modified protocol (cDNA amplification method). The results showed that cDNA amplification method provided high reproducibility, representation, and concordance with the standard cDNA non-amplification method. We have successfully analyzed the gene expression profiles of needle biopsy tissues using the modified method without significantly changing the expression profiles. These results suggest that the global gene expression profiles of small biopsy samples can be achieved by our modified method to facilitate the analysis of gene expression profiles for clinical application.

Biopsy, Needle↗
PubMed1998

Fenofibrate modifies transaminase gene expression via a peroxisome proliferator activated receptor alpha-dependent pathway.

Fibrates modify the expression of genes implicated in lipoprotein and fatty acid metabolism via the peroxisome proliferator-activated receptor alpha(PPARalpha), leading to reductions in serum triglycerides and cholesterol. The expression of certain genes regulated by PPARalpha have been shown to be modified in a species dependent manner. Aspartate aminotransferase (AspAT or GOT) and alanine aminotransferase (AlaAT or GPT) are enzymes involved in intermediate metabolism in all cells and in hepatic gluconeogenesis. These enzymes are also widely used as serum markers of possible tissue damage. This study investigated whether fenofibrate could modify the expression of liver AspAT and/or AlaAT and thus possibly alter transaminase levels independently of a cytotoxic effect. In human Hep G2 cells, fenofibrate increased cytosolic AspAT (cAspAT) activity by 40% and AlaAT activity by 100%, as well as both mRNAs. Nuclear run on assays showed that this effect was, at least in part, transcriptional. Increases in mRNA were also observed in human hepatocyte cultures at concentrations of the drug attained in patients. In C57BL/6 mice, fenofibrate decreased cAspAT and cAlaAT mRNA, while these effects were abolished in PPARalpha knock-out mice. In conclusion, fenofibrate has been shown to modify cAspAT and AlaAT gene expression in a species and PPARalpha dependent manner. This is the first demonstration that cAspAT and AlaAT activities may be pharmacologically altered, independently of a toxic phenomenon.

Alanine Transaminase↗
PubMed1998

Disposable, enzymatically modified printed film carbon electrodes for use in the high-performance liquid chromatographic-electrochemical detection of glucose or hydrogen peroxide from immobilized enzyme reactors.

Disposable screen-printed, film carbon electrodes (PFCE) were modified with cast-coated Osmium-polyvinylpyrridine-wired horse radish peroxidase gel polymer (Os-gel-HRP) to enable the detection of the reduction at 0 mV of hydrogen peroxide (H2O2) derived from a post-column immobilized enzyme reactor (IMER) containing acetylcholinesterase and choline oxidase. In another series of experiments PFCE were initially modified with cast-coated Os-gel-HRP and then treated with glucose oxidase in bovine serum albumin (BSA) and cross-linked with glutaraldehyde to form a bi-layer glucose-Os-gel-HRP PFCE. This bi-layer glucose-Os-gel-HRP PFCE generated a reduction current at 0 mV to H2O2 derived from the reaction of glucose oxidase and glucose in solution. These enzyme-modified PFCE were housed in a radial flow cell and coupled with cation-exchange liquid chromatographic methods to temporally separate substrates in solution for the determination of acetylcholine (ACh) and choline (Ch) in the first experimental series, or glucose in the second experimental series. These two disposable enzyme-modified PFCE exhibited linear current vs. substrate relations, were durable, being usable for approximately 40 determinations, and were sufficiently sensitive to be employed in biological sampling. Both assays utilized the same HPLC equipment. The limit of detection for ACh was 16 fmol/10 microl and that for glucose was 12 micromol/7.5 microl. ACh and Ch were measured from a microdialysate from the frontal cortex of a rat. Glucose in human urine was determined using the bi-layer glucose oxidase-Os-gel-HRP PFCE.

Acetylcholine↗
PubMed1998

Development of tryptophan-modified human serum albumin columns for site-specific studies of drug-protein interactions by high-performance affinity chromatography.

Human serum albumin (HSA) is one of the main proteins involved in the binding of drugs and small solutes in blood or serum. This study examined the changes in chromatographic properties that occur for immobilized HSA following the chemical modification of HSA's lone tryptophan residue (Trp-214). Trp-214 was reacted with o-nitrophenylsulfenyl chloride, followed by immobilization of the modified protein and normal HSA onto separate silica-based HPLC supports. The binding properties of the modified and normal HSA were then analyzed and compared by using frontal analysis and zonal elution experiments employing R/S-warfarin and L-tryptophan as probe compounds for the warfarin and indole binding regions of HSA. The modified HSA was found to have the same number of binding sites as normal HSA for R-warfarin and L-tryptophan but lower association equilibrium constants for these test solutes. Zonal elution studies with R- and S-warfarin on the modified HSA column demonstrated the importance of Trp-214 in determining the stereoselective binding of HSA for these agents. These studies also indicated that tryptophan modification can alter HSA-based separations for chiral solutes.

Apazone↗
PubMed1999

Preparation of poly(DL-lactide-co-glycolide) nanoparticles by modified spontaneous emulsification solvent diffusion method.

PURPOSE: The objectives of this study were to establish a new preparation method for poly(DL-lactide-co-glycolide) (PLGA) nanoparticles by modifying the spontaneous emulsification solvent diffusion (SESD) method and to elucidate the mechanism of nanoparticle formation on the basis of the phase separation principle of PLGA and poly(vinyl alcohol) (PVA) in the preparation system. METHODS: PLGA nanoparticles were prepared by the modified-SESD method using various solvent systems consisting of two water-miscible organic solvents, in which one solvent has more affinity to PLGA than to PVA and the other has more affinity to PVA than to PLGA. The yield, particle size, size distribution and PVA content of the PLGA nanoparticles were evaluated, and the phase separation behaviors of the polymers were elucidated. RESULTS: The modified-SESD method provided a good yield of PLGA nanoparticles over a wide range of composition ratios in the binary mixture of organic solvents. Several process parameters, including the fed amount of PLGA, PLGA concentration and PVA concentration were examined to achieve the optimum preparation conditions. The discrete powder of PLGA nanoparticles was obtained by freeze-drying. No change in the PVA content of PLGA nanoparticles was observed even after several times of washing treatment by ultrafiltration, suggesting a strong surface adsorption. It was found that the appropriate selections of binary solvent mixtures and polymeric concentrations in both organic and aqueous phases could provide excellent yield and favorable physical properties of PLGA nanoparticles. CONCLUSION: The proposed modified-SESD method can be used to provide PLGA nanoparticles of satisfactory quality at an acceptable yield for industrial purposes.

Diffusion↗
PubMed2003

Flow injection analysis of ascorbic acid using carriers modified with metal-porphine as oxidative solid catalyses.

To investigate one of practical applications of the supports modified with metal-porphines as artificial solid-catalysts, columns into which the supports were packed were supplied to catalytic columns for a flow injection analysis (FIA) system for determination of ascorbic acid (AsA) by the following reactions: AsA+O(2)-->dehydoroAsA+H(2)O(2),H(2)O(2)+chromogen-->2H(2)O+Dye. Among the columns tested, the column containing silica gels modified with Co-tetrakis(carboxyphenyl)porphine catalyzed most rapidly the oxidation reaction of AsA that is accompanied by the formation of hydrogen peroxide. The resulting hydrogen peroxide was determined by FIA system equipped with the column containing glass beads modified with Mn-tetrakis(carboxyphenyl)porphine, which gave a linear calibration curve and large peak-areas of the range corresponding to AsA concentration between 0.2 and 10 micromol/ml. The results indicated that some supports modified with metal-porphine would be applicable to the FIA for AsA as the solid catalyses which function as if the immobilized enzymes.

Ascorbic Acid↗
PubMed2003

Modified resolution factor for asymmetrical peaks in chromatographic separation.

The quality of separation is measured by resolution factor (R(s)) between adjacent peaks. The current United States Pharmacopeia (USP) method for R(s) calculation assumes symmetrical peak shapes. In practice, perfectly symmetrical peaks are rarely encountered. The goal of this study was to correct the inaccuracy due to peak asymmetry by using a new formula for R(s) calculation. Peak tailing factor was incorporated into the formula for the calculation of R(s) to correct for the peak asymmetry. The resulting modified formula was compared with USP formula using simulated peaks and actual peaks. Through mathematical derivation, the modified R(s) was expressed as: R=[2(t(2)+W(2)/2(1-1/TF(2))-t(1)-W(1)/2(1-1/TF(1)))]/W(1)+W(2) where t(1) and t(2) are the retention times of two peaks, W(1) and W(2) are peak widths at baseline, TF(1) and TF(2) are USP tailing factors. All parameters used in the formula are available from commercial data analysis software. Comparisons of modified R(s) to USP R(s) showed that the modified R(s) provided a more accurate measure of peak separation at the baseline level.

Chromatography, High Pressure Liquid↗
PubMed1997

The influence of temperature on the multiple separation of estrogenic steroids using mobile phases modified with beta-cyclodextrin in high-performance liquid chromatography.

The effect of temperature on the retention and multiple separation of six estrogenic steroids in reversed-phase liquid chromatography has been studied. Capacity factors (k') of estriol, 17 beta-estradiol, 17 alpha-estradiol, d-equilenin, equilin and estrone were measured using mobile phase modified with different concentrations of beta-cyclodextrin (from 0-16 mM), a fixed solvent composition (acetonitrile-water) and a wide range of column temperatures (from 5 to 80 degrees C). The plots of capacity factors vs. reciprocal of absolute temperature are nonlinear in each case when mobile phase modified with beta-cyclodextrin was used. Particularly strong nonlinearity was observed at lower temperature and at higher beta-cyclodextrin concentration. The complex chromatograms were evaluated using optimization parameters such as capacity factor of the last-eluted peak (k'max), the smallest resolution between adjacent peaks (Rs,min) and relative resolution product (r). The results presented describe precisely the role of temperature in high-performance liquid chromatography systems in which mobile phases modified with cyclodextrin were used. Moreover, the elution order of estrogenic steroids on modified and unmodified mobile phases has been discussed.

Chromatography, High Pressure Liquid↗
PubMed2003

Yeast cell adhesion on oligopeptide modified surfaces.

Self-assembling oligopeptides are novel materials with potential bioengineering applications; this paper explores the use of one of these oligopeptides, EAK 16 II, for modifying the surface properties of cell-supporting substrates. To characterize the surface properties, thermodynamic measurements of liquid contact angle and surface free energy were correlated to atomic force microscopy (AFM) observations. A critical concentration of 0.1 mg/ml was found necessary to completely modify the surface properties of the substrate with EAK 16 II. Adhesion of a yeast cell, Candida utilis, was modified by the coating of EAK 16 II on both hydrophobic (plastic) and hydrophilic (glass) surfaces: Cell coverage was slightly enhanced on the glass substrate, but decreased significantly on the plastic substrate. This indicates that the yeast cell adhesion was mainly determined via hydrophobic interactions between the substrate and the cell wall. However, on the EAK 16 II modified glass substrate, surface roughness might be a factor in causing a slightly larger cell adhesion than that on bare glass. The morphology of adhered cells was also obtained with AFM imaging, showing a depression at the center of the cell on all substrates. Small depressions on the oligopeptide-coated surfaces and plastic substrate may indicate good water-retaining ability by the cell. There was no apparent difference in cell adhesion and morphology among cells obtained from lag, exponential and stationary growth phases.

Candida↗
PubMed2001

Enhanced matrix synthesis and in vitro formation of cartilage-like tissue by genetically modified chondrocytes expressing BMP-7.

Bone morphogenic protein-7 (BMP-7) supports ectopic cartilage and bone formation, is expressed in normal articular cartilage, and increases matrix synthesis in chondrocytes. Based on this knowledge, we hypothesized that an adenovirus (Ad) vector encoding human BMP-7 could be used to modify chondrocytes genetically to improve their capacity for cartilage repair. An adenovirus vector encoding BMP-7 (AdBMP-7) was constructed and its bioactivity confirmed by ectopic bone formation assay. AdBMP-7 modification of bovine chondrocytes induced expression of BMP-7 mRNA and bioactive protein, resulting in an increase in incorporation of 35SO4- into proteoglycan, 3H-proline uptake into protein, and the expression of the cartilage-specific matrix genes, aggrecan and type II collagen. An in vitro model of chondrocyte transplantation was used to demonstrate the feasibility of using genetically modified chondrocytes to enhance formation of cartilage-like tissue. When transplanted onto cartilage explants and maintained in vitro for 3 weeks, chondrocytes modified with AdBMP-7 formed 1.9-fold thicker tissue than chondrocytes modified with a control vector (P < 0.001). This tissue was positive for type II collagen and proteoglycan but negative for type X collagen and demonstrated a cartilage-like morphology. These observations suggest that Ad-mediated transfer of BMP-7 gene to chondrocytes enhances the chondrocyte-specific matrix synthesis and their capacity to form cartilage-like tissue, thus representing a strategy that may improve cell-based cartilage repair.

Adenoviridae↗
PubMed2003

Modifiers of gastrointestinal motility of cattle.

Little clinically relevant, evidence-based data about the effect of motility modifiers on the GI tract of cattle are currently available. Additionally, some of the published results seem to be contradictory. Three main facts explain this apparent discrepancy: (1) Results may not be transferred from one species to another, because presence, concentration, location, distribution, and function of specific receptors may differ significantly among species. (2) The lack of a significant effect on smooth muscle preparations in vitro does not necessarily exclude a certain drug's motility-modifying property on affected animals in vivo. Certain drugs bind to receptors outside the myenteric plexus. Others, such as lidocaine and adrenergic-, dopaminergic-, and opioid-antagonists increase only GI motility, if inhibitory reflexes or a hyperactive state of the inhibitory sympathetic nervous system preexists. (3) Effects of motility modifiers as found in healthy experimental animals in vivo may not be similar to those found in spontaneously diseased animals. Accurate and reliable data on the effect of modifiers of GI motility of cattle will be obtained only from double-blinded, evidence-based, in vivo studies on spontaneously affected animals. Because well-documented results from such studies are extremely rare, intensive research in this field is warranted in the future.

Animals↗
PubMed1992

Inflammatory response in the rabbit eye after intraocular implantation with poly(methyl methacrylate) and heparin surface modified intraocular lenses.

Early inflammatory responses in rabbit eyes after anterior chamber implantation with poly(methyl methacrylate) (PMMA) and heparin surface modified intraocular lenses (IOLs) and after cataract extraction and implantation with PMMA and heparin surface modified intraocular lenses (IOLs) were investigated between 1 and 30 days postoperatively. The number of white blood cells in the aqueous humor, the distribution of white blood cell subsets, and the interleukin-1 (IL-1b) levels were studied. At one day postoperatively, there was a significantly smaller number of white blood cells in eyes with heparin surface modified IOLs than in eyes with PMMA IOLs after both anterior chamber implantation and cataract surgery and capsular bag implantation. At one day postoperatively, 87% of the cell population in eyes with PMMA IOLs were neutrophilic granulocytes. Two days later the neutrophilic granulocytes had decreased to 13% and at seven days they were gone. At 14 days the frequency was 7% and at 30 days it was 27%. The fraction of monocytes/macrophages in the cell population was 9% (day 1), 79% (day 3), 94% (day 7), 81% (day 14), and 72% (day 30). The fraction of lymphocytes was low throughout the test period (1% to 10%). The percentages were almost the same in eyes with heparin surface modified IOLs and there was no significant difference in the distribution of white blood cell subsets between the two lens materials. The IL-1 was not detected in any of the samples (day 3, day 7, and day 30). Inactivated macrophages and/or levels under detection limits could be the reason.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2001

Peroxynitrite-modified 99mTc-beta-VLDL: tissue distribution and plasma clearance rate.

Free radicals superoxide (O(2)(-)) and nitric oxide (*NO) are generated by blood vessels and can rapidly react to produce a peroxynitrite anion (ONOO(-)), a powerful oxidant that modifies lipoproteins making them more atherogenic. The aim of this study was to investigate the effect of peroxynitrite-induced modifications on beta-very-low-density lipoprotein (beta-VLDL) as to its biodistribution and plasma clearance rate, as well as the uptake of these particles by THP-1 cells. After being injected into New Zealand White rabbits, the peroxynitrite-modified beta-VLDL (99mTc-per-beta-VLDL) was cleared from circulation faster than the native beta-VLDL (99mTc-nat-beta-VLDL) in both normocholesterolemic rabbits (NC) and in hypercholesterolemic rabbits (HC). In HC rabbits, the fractional clearance of 99mTc-labeled beta-VLDL was significantly lower than in NC rabbits. The in vivo studies showed that accumulation of 99mTc-labeled beta-VLDL, expressed per gram of tissue, followed the decreasing order: kidney > liver > spleen > adrenal gland >or= lung > aortic arch > heart >or= abdominal aorta > thoracic aorta > psoas muscle. The high accumulation in the kidneys suggests the processing of 99mTc-labeled apolipoproteins by receptors present in kidney cells. The accumulation of 99mTc-nat-beta-VLDL in the whole organ was the following: liver > kidney > heart > spleen > adrenal gland > aorta in HC and NC rabbits. The uptake of 99mTc-per-beta-VLDL by the spleen was greater than the uptake by the heart in both groups. The in vitro studies showed that the uptake of 99mTc-per-beta-VLDL by THP-1 cells was higher than that of 99mTc-nat-beta-VLDL. These results show that peroxynitrite-modified beta-VLDL is rapidly removed from plasma and accumulates in several tissues, mainly in the liver and kidney. This may be particularly important in hypercholesterolemic situations that could favor the accumulation of native and peroxynitrite-modified beta-VLDL in several tissues.

Animals↗
PubMed2002

Carbonyl modified proteins in cellular regulation, aging, and disease.

The oxidative modification of proteins by reactive species is implicated in the etiology or progression of a panoply of disorders and diseases. The level of these modified molecules can be quantitated by measurement of the protein carbonyl content, which has been shown to increase in a variety of diseases and processes, notably during aging. For the most part, oxidatively modified proteins are not repaired and must be removed by proteolytic degradation, a process which normally proceeds very efficiently, from microorganisms to mammals. In eukaryotes, removal is usually carried out by the proteosome, which selectively degrades oxidatively modified proteins, whether they be damaged by reactive oxygen species or specifically oxidized by cellular regulatory processes. The molecular deficiencies that cause accumulation of oxidatively modified proteins are not identified, but regardless of cause, the accumulation is likely to disrupt normal cellular function.

Aging↗
PubMed1997

Increased levels of 4-hydroxynonenal modified proteins in plasma of children with autoimmune diseases.

Analysis of serum samples of healthy children (n = 11) and children with Systemic Lupus Erythematosus (SLE), (n = 21) was performed by SDS-PAGE and immunoblot with an antibody directed against proteins modified by lipid peroxidation (LPO) product 4-hydroxynonenal (HNE). A single major stained protein band was detected. By comparison of the molecular weights in nonreducing and reducing SDS-PAGE was found that the main protein modified by HNE is immunoglobulin G. Significantly higher concentrations of the aldehyde modified protein were found in children with high disease activity of SLE measured by SLE disease activity index (SLEDAI). Lipid peroxidation measured by malondialdehyde and 4-hydroxynonenal concentrations show an enhanced level of both compounds also in patients with the active autoimmune disease. Therefore, it can be assumed that free radical mediated processes play a pathophysiological role in the active phase of SLE and HNE-modified serum proteins are a further parameter for the detection of in vivo LPO.

Adolescent↗
PubMed2003

The increased bactericidal activity of a fatty acid-modified synthetic antimicrobial peptide of human cathepsin G correlates with its enhanced capacity to interact with model membranes.

The bactericidal potency of a synthetic peptide (CG 117-136) of human lysosomal cathepsin G (cat G) can be substantially increased by covalent attachment to its N- or C-termini, of saturated, linear fatty acids (FAs), namely those with C-8, C-10 and C-12 hydrocarbon chains. In order to understand better the mechanism by which FA moieties increase the bactericidal activity of CG 117-136, the interaction of N-terminally FA-modified peptides with artificial membranes was studied. First, the content of secondary structure motifs in the modified and unmodified peptides was determined by circular dichroism (CD). A marked increase in the propensity of FA-modified CG 117-136 to form an alpha-helix structure was observed for the C-8, C-10 and C-12 derivatives compared with unmodified/short-chain and long-chain (C-14, C-16, C-18) derivatives. These effects were observed both in the presence of large unilamellar liposomes or in trifuluoroethanol, a membrane-stimulating agent. Second, the capacity of peptides to insert into large unilamellar liposomes as a function of FA length was determined by their ability to release a trapped fluorescent dye. FA derivatives with the highest alpha-helical content were found to be the most effective in releasing a fluorescent dye, compared with an unmodified peptide and/or derivatives having a low alpha-helical content. The ability of the peptides to attain alpha-helical structure in the membrane-like environment and the ability to disrupt the liposomal membrane, therefore correlate remarkably well with their increased ability to kill bacteria. A plausible explanation for improved bactericidal action of the modified peptide is that the FA moiety facilitates formation of the peptide with an alpha-helical structure formation in membranes, which is essential for disrupting the integrity of the bacterial cytoplasmic membrane.

Amino Acid Sequence↗
PubMed2000

Detection of genes encoding aminoglycoside-modifying enzymes in staphylococci by polymerase chain reaction and dot blot hybridization.

Dot blot hybridization and the polymerase chain reaction (PCR) were used to study aminoglycoside-modifying enzymes in aminoglycoside-resistant staphylococci isolated in hospitals in Kuwait. DNA encoding the acetyltransferase (AAC) (6')-phosphotransferase (APH) (2"), nucleotidyltransferase (ANT) (4') and APH (3') enzymes were detected in Staphylococcus aureus and coagulase negative staphylococci. ANT (4') was the most common enzyme detected. The majority of isolates contained genes for all three modifying enzymes, AAC (6')-APH (2"), ANT (4') and APH (3'); only few isolates carried genes for a single modifying enzyme. Genes encoding the AAC (6')-APH (2") were detected in all except two gentamicin-resistant isolates. In these isolates the genes for the AAC (6')-APH (2") enzyme could not be detected by PCR and dot blot hybridization. Whereas antibiotic resistance testing could be used to predict the presence of the AAC (6')-APH (2") enzyme it was not useful in predicting the presence of the ANT (4') or APH (3') enzymes in gentamicin-resistant isolates. Results obtained with dot blot hybridization were comparable to those obtained with PCR. However, PCR was fast and results were obtained within the same day. Therefore PCR would be preferred for the detection and confirmation of the presence of aminoglycoside-modifying enzymes in clinical microbiology laboratories.

Acetyltransferases↗