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PubMed1992

Selective determination of arginine-containing and tyrosine-containing peptides using capillary electrophoresis and laser-induced fluorescence detection.

The use of two different amino acid-selective fluorogenic reagents for the derivatization of peptides is investigated. One such scheme utilizes a selective reaction of benzoin with the guanidine moiety to derivatize arginine residues occurring in a peptide. The second scheme involves the formylation of tyrosine, followed by reaction with 4-methoxy-1,2-phenylenediamine. The use of capillary electrophoresis and laser-induced fluorescence detection allows enhanced efficiencies and sensitivities to be obtained for the separations of either arginine- or tyrosine-containing peptides. A helium-cadmium laser (325 nm) is ideally suited for the laser-based detection system due to a close match of the excitation maxima of derivatized peptides from both reactions. A detection limit of 270 amol is achieved for model arginine-containing peptides, while the detection limit for model tyrosine-containing peptides is measured at 390 amol. Both derivatization reactions are found to be useful for high-sensitivity peptide mapping applications in which only the peptides containing the derivatized amino acids are detected.

Amino Acid Sequence↗
PubMed1985

Metabolism of phosphorus-containing compounds by pig liver microsomal FAD-containing monooxygenase.

Oxidative desulfuration of phosphonate insecticides such as fonofos (S-phenyl ethyl ethylphosphonodithioate) and its analogs is catalyzed by pig liver microsomal FAD-containing monooxygenase, although desulfuration of phosphorodithioates, such as parathion, is not. Substitution of an alkyl group for the remaining alkoxy group, as in S-phenyl diethylphosphinodithioate, did not increase its oxidation rate. Diethylphenylphosphine sulfide, containing 3 phosphorus-carbon bonds, was actually a poorer substrate than fonofos. Replacement of the S-phenyl group of fonofos with an O-phenyl group increased the Km value above the solubility limit. Trivalent phosphorus-containing compounds were also excellent substrates for this enzyme. Diethylphenylphosphine had a Km value lower than 2.5 microM. Diethyl phenylphosphonite also appeared to be an excellent substrate but its rapid nonenzymatic hydrolysis and/or oxidation precluded accurate Km determinations. Stoichiometry studies with diethylphenylphosphine and its sulfide showed that O2 and NADPH consumption were approximately equal to the substrate consumed. The major metabolite of both diethylphenylphosphine and its sulfide was the phosphine oxide. These results show that microsomal FAD-containing monooxygenase of pig liver has activity as a phosphorus-oxidase, in addition to its well characterized nitrogen- and sulfur-oxidase roles.

Animals↗
PubMed1993

Galanin-containing fibers innervate substance P-containing neurons in the pedunculopontine tegmental nucleus in humans.

Galanin is a putative peptide transmitter distributed widely in the central nervous system. Galanin shows potent inhibitory effects on neuronal activity and learning behaviors induced or mediated by acetylcholine. Here, we demonstrate that in humans there are abundant galanin-containing fibers in the pedunculopontine tegmental nucleus, the lateral dorsal tegmental nucleus and the oral pontine reticular nucleus. These neurons contain both acetylcholine and substance P (SP). Using a double-immunostaining method, we show that galanin-containing terminals from pericellular baskets around many SP-positive neurons in these nuclei. Our results suggest galanin may influence brain functions via inputs to the ascending mesopontine acetylcholine-containing nuclei.

Aged↗
PubMed1986

The antiseptic efficacy of chlorxylenol-containing vs. chlorhexidine gluconate-containing surgical scrub preparations.

The studies described here evaluate the efficacy of the chlorxylenol-containing surgical scrub formulations against the chlorhexidine gluconate-containing formulations using the Glove Juice Test, as recommended by the FDA's panel to develop guidelines for the study of antiseptic agents. Similar reports from the literature evaluating the relative efficacies of the iodophor-containing and the hexachlorophene-containing formulations are cited. Results fail to detect any significant differences in the efficacy of these two preparations, each significantly reducing the bacterial flora on the hands as indicated by immediate post-wash colony counts, and each demonstrating the continuing ability to significantly reduce bacterial growth with continued regular use.

Anti-Infective Agents, Local↗
PubMed2004

Structural determination of five novel tetrasaccharides containing 3-O-sulfated D-glucuronic acid and two rare oligosaccharides containing a beta-D-glucose branch isolated from squid cartilage chondroitin sulfate E.

Oversulfated chondroitin sulfate E (CS-E) derived from squid cartilage exhibits intriguing biological activities, which appear to reflect the biological activities of mammalian CS chains containing the so-called E disaccharide unit [GlcAbeta1-3GalNAc(4,6-O-disulfate)]. Previously, we isolated novel tetra- and hexasaccharides containing a rare GlcA(3-O-sulfate) at the nonreducing end after digestion of squid cartilage CS-E with testicular hyaluronidase. In this study, squid cartilage CS-E was extensively digested with chondroitinase AC-II, which yielded five highly sulfated novel tetrasaccharides and two odd-numbered oligosaccharides (tri- and pentasaccharides) containing D-Glc. Their structures were determined by fast atom bombardment mass spectrometry and (1)H NMR spectroscopy. The results revealed an internal GlcA(3-O-sulfate) residue for all the novel tetrasaccharide sequences, which rendered the oligosaccharides resistant to the enzyme. The results suggest that GlcA(3-O-sulfate) units are not clustered but rather interspersed in the CS-E polysaccahride chains, being preferentially located in the highly sulfated sequences. The predominant structure on the nearest nonreducing side of a GlcA(3-O-sulfate) residue was GalNAc(4-O-sulfate) (80%), whereas that on the reducing side was GalNAc(4,6-O-disulfate) (59%). The structural variety in the vicinity of the GlcA(3-O-sulfate) residue might represent the substrate specificity of the unidentified chondroitin GlcA 3-O-sulfotransferase. The results also revealed a trisaccharide and a pentasaccahride sequence, both of which contained a beta-d-Glc branch at the C6 position of the constituent GalNAc residue. Approximately 5 mol % of all disaccharide units were substituted by Glc in the CS-E preparation used.

Animals↗
PubMed1983

Synergy of ricin A chain-containing immunotoxins and ricin B chain-containing immunotoxins in in vitro killing of neoplastic human B cells.

A strategy is described for improving the efficacy of ricin A chain-containing immunotoxins. Highly purified preparations of ricin A chain or ricin B chain were separately coupled to anti-human immunoglobulin antibodies and the conjugates (immunotoxins) were affinity purified to eliminate free chains. Mixtures of anti-Ig-A and anti-Ig-B immunotoxins markedly synergized in vitro in their ability to kill the Ig-bearing human lymphoma cell line Daudi. In contrast, A-chain- or B-chain-containing immunotoxins of irrelevant specificity did not synergize with anti-Ig-A or anti-Ig-B immunotoxins. This finding indicates that free A or B chains do not play a major role in the synergy and that the synergy is specific. Thus, synergy depends on the specificity of the two antibodies; the lectin binding ability of a B-chain-containing immunotoxin of irrelevant specificity does not suffice. This approach of delivering ricin A chain and B chain separately to a target cell may have significant advantages in killing cells that are not effectively killed by A-chain-containing immunotoxins alone.

Burkitt Lymphoma↗
PubMed1990

Reduced selenium status in chicks given diets low in sulphur-containing amino acids and containing sodium nitroprusside as a source of cyanide.

1. Two experiments were carried out to study the effect of dietary cyanide, given in the form of sodium nitroprusside (SNP), on the growth and selenium status of chicks fed on diets low in sulphur-containing amino acids. 2. In experiment 1, SNP (0.3 g/kg) depressed growth rate and food intake when included in diets low in total sulphur-containing amino acids. It also reduced plasma and liver glutathione peroxidase activity (GSH-Px) and kidney selenium concentration. These latter variables also showed increases in response to supplements of selenium of 0.1 and 0.2 mg selenium/kg (as sodium selenite) although growth was not affected. 3. In experiment 2, SNP did not affect growth when reduced to 0.1 g/kg in diets low both in methionine and total sulphur-containing amino acids. It did, however, reduce plasma GSH-Px activity, which again increased in response to similar dietary supplements of selenium. 4. It is concluded that dietary cyanide effectively increases the requirement for selenium and could induce deficiency in diets only marginally adequate in selenium, particularly when the supply of sulphur-containing amino acids was marginal or inadequate.

Amino Acids, Sulfur↗
PubMed2001

The influence of excipients on the stability of the moisture sensitive drugs aspirin and niacinamide: comparison of tablets containing lactose monohydrate with tablets containing anhydrous lactose.

The purpose of this study is to test the hypothesis that in tablet formulations, moisture-sensitive drugs formulated with lactose monohydrate have the same stability as formulations containing anhydrous lactose, and to characterize the kinetics of niacinamide degradation in the solid state. Aspirin and niacinamide decomposition were used as indicators of stability. Aspirin and niacinamide tablets containing either lactose monohydrate or anhydrous lactose were separately investigated at different temperatures and relative humidities; the stability tests were done at 25 degrees C--60% RH, 40 degrees C--80% RH, 60 degrees C--60% RH, 60 degrees C--80% RH, and 80 degrees C--80% RH. Official U.S. Pharmacopeia methods were used for the aspirin and niacinamide assays. Statistical analysis showed that tablets containing lactose monohydrate have the same stability as tablets containing anhydrous lactose, which means that even though water is present in the crystal structure, the bound water does not influence the reaction rate. In addition, niacinamide degradation in the solid-state can be described by a third order rate equation.

Aspirin↗
PubMed1999

Efficacy of salvage therapy containing ritonavir and saquinavir after failure of single protease inhibitor-containing regimens.

OBJECTIVE: To assess the efficacy of salvage therapy containing ritonavir and saquinavir after failure of indinavir- or nelfinavir-containing regimens, and to determine correlates of success or failure. DESIGN: Retrospective chart review. SETTING. The Moore Clinic - the HIV clinic of Johns Hopkins Hospital. PATIENTS: Forty-one HIV-infected patients were identified through physician contacts, referrals from other providers, and review of a comprehensive clinical database. MAIN OUTCOME MEASURES: To determine response to salvage therapy, HIV-1 viral RNA (absolute and log10-transformed) was measured using the Roche Amplicor quantitative HIV-1 RNA assay after initiation of the salvage regimen. Potential correlates of response included: viral RNA at the time of switch; viral RNA at the time of switch as a percentage of baseline viral RNA; magnitude of decline in viral RNA; and the interval between virologic failure of single protease inhibitor therapy and switch to the salvage regimen. RESULTS: Thirteen (56.5%) of 23 patients failing indinavir responded to salvage therapy (HIV RNA < 400 copies/ml) with persistence throughout the follow-up period (median of 37 weeks; range 18-67 weeks). Mean absolute viral RNA at the time of switch was 20 238 copies/ml (median, 9281) compared with 42 953 copies/ml (median, 24 650) for the 10 non-responders. Mean log10 viral RNA at switch was 3.804 for responders versus 4.405 for non-responders (P = 0.040). Among four responders who had failed nelfinavir, mean viral RNA was 9634 copies/ml and mean log10 viral RNA was 3.749 at the time of switch. Two non-responders had a mean viral RNA of 21 551 and a mean log10 viral RNA of 4.037 at switch. CONCLUSIONS: In contrast with previous reports, salvage regimens containing ritonavir and/or saquinavir can be effective and durable following the failure of combination regimens containing either indinavir or nelfinavir. Salvage therapy may be more likely to succeed when it is initiated early in failure at low viral loads.

Adult↗
PubMed2001

Comparison of virologic, immunologic, and clinical response to five different initial protease inhibitor-containing and nevirapine-containing regimens.

CONTEXT: The effectiveness of different protease inhibitors (PIs) and non-nucleoside reverse transcriptase inhibitors outside the setting of clinical trials has not been well described. OBJECTIVES: To compare five different PI-and nevirapine (NVP)-containing regimens on virologic, immunologic, and clinical outcomes and treatment discontinuation. DESIGN AND SETTING: Observational cohort study based on an HIV clinic in London. PATIENTS: A total of 690 patients who received either saquinavir hard gel (SQV HG) (n = 183), indinavir (IDV) (n = 189), nelfinavir (NFV) (n = 109), ritonavir (RTV) (n = 42), ritonavir with saquinavir hard gel (RTV/SQV HG) (n = 45), or NVP (n = 122) as part of an initial PI-or NVP-containing treatment regimen between November 1994 and December 1998. A total of 351 (51%) patients had prior exposure to nucleoside reverse transcriptase inhibitors (NRTIs). MAIN OUTCOME MEASURES: The main outcome measures were virologic undetectability, subsequent virologic rebound, CD4 cell count rise, development of AIDS, and treatment discontinuation. All analyses were stratified for year of initiation of the PI-or NVP-containing regimen. RESULTS: Overall, 63% of patients attained an undetectable viral load (VL) within 6 months of starting their PI or NVP regimen. The adjusted relative hazard (95% confidence interval [CI]) for an undetectable VL relative to SQV HG was (in rank order): 2.77 (CI: 1.84-4.17) for NFV, 2.54 (CI: 1.81-3.57) for IDV, 2.43 (CI: 1.52-3.87) for RTV, 2.08 (CI: 1.28-3.37) for RTV/SQV HG, and 1.96 (CI: 1.35-2.85) for NVP. Forty-nine percent of patients experienced VL rebound within 12 months of initial attainment of undetectability, but relative to SQV HG, this did not differ significantly across the different PI and NVP regimens. The CD4 cell count response and rate of AIDS events were also similar across the different regimens. No independent predictors of VL undetectability were identified, but prior NRTI exposure was associated with VL rebound, and a lower baseline VL and CD4 cell count were associated with a reduced CD4 count response. The frequency (95% CI) of treatment discontinuation differed across the regimens; at 6 months, it was lowest for NFV (18% [CI: 13%-24%]), IDV (25% [CI: 22%-29%]), and NVP (28% [CI: 22%-34%]) and highest for RTV (41% [CI: 31%-52%]) and SQV HG (52% [CI: 48%-57%]). CONCLUSIONS: Although PI- and NVP-containing regimens were similar in their CD4 cell count response and rates of subsequent VL rebound, differences were observed in time to VL undetectability and discontinuation rates relative to SQV HG. SQV HG was consistently inferior to the other PIs and NVP. The use of NFV and IDV was associated with the highest rates of undetectability, and together with NVP, the lowest rates of discontinuation.

Adult↗
PubMed1989

Effects of bicarbonate-containing versus lactate-containing peritoneal dialysis solutions on superoxide production by human neutrophils.

Human neutrophils were isolated from healthy volunteers and exposed to either bicarbonate-containing (pH 7.4) or lactate-containing (pH 5.2) peritoneal dialysis solution in vitro. Superoxide production by neutrophils was measured by a method based on the superoxide dismutase-inhibitable reduction of ferricytochrome c. Bicarbonate-containing peritoneal dialysis solution was found to be superior to the lactate-containing one in facilitating the production of superoxide anion by human neutrophils.

Adult↗
PubMed2003

The effect of a toothpaste containing 2% zinc citrate and 0.3% Triclosan on bacterial viability and plaque growth in vivo compared to a toothpaste containing 0.3% Triclosan and 2% copolymer.

OBJECTIVE: To compare the antimicrobial efficacy and effect on plaque growth of a new silica-based fluoride toothpaste containing 2% zinc citrate/ 0.3% Triclosan with a silica-based fluoride toothpaste containing 0.3% Triclosan/2% copolymer. METHODS: In Study 1, plaque was collected after one week's use of each toothpaste and assessed for bacterial viability, live/ dead ratio and microbial membrane integrity. In study 2, plaque was measured immediately and 18 hours after a single brushing with the specified toothpastes. RESULTS: The 2% zinc citrate/0.3% Triclosan formulation significantly reduced the total number of viable aerobic and anaerobic bacteria (p = 0.0223 and p = 0.0443 respectively) compared to the 0.3% Triclosan/2% copolymer formulation. Both toothpastes increased the bacterial membrane permeability significantly. However, the proportion of live bacteria for the 2% zinc citrate/0.3% Triclosan product was significantly reduced (p < 0.05). Study 2 showed significantly less plaque growth 18 hours after using the 2% zinc citrate/0.3% Triclosan toothpaste compared to the 0.3% Triclosan/2% copolymer toothpaste (p < 0.01). CONCLUSION: Regular use of a fluoride toothpaste containing 2% zinc citrate and 0.3% Triclosan, significantly reduced the viability of plaque bacteria compared to a fluoride toothpaste containing 0.3% Triclosan/ 2% copolymer 12 hours after brushing. In addition, a clinical plaque growth study confirmed that this anti-microbial efficacy leads to a significant reduction in plaque growth.

Adult↗
PubMed1984

Serum-resistant mutants of Escherichia coli O111 contain increased lipopolysaccharide, lack an O antigen-containing capsule, and cover more of their lipid A core with O antigen.

Escherichia coli strains of group O111 were characterized with respect to sensitivity to complement killing, amount of lipopolysaccharide and O antigen-containing capsule, and distribution of O antigen. All wild-type E. coli O111 strains were resistant to complement killing in the absence of specific antibody. Presensitization of strains with antibody to whole cells (OK antibody), followed by incubation in 50% pooled normal human serum as a source of complement, subdivided wild-type strains into three types: completely resistant, partially resistant, and sensitive. Completely and partially resistant mutants were isolated by cycles of serum killing, starting with one sensitive strain. Completely resistant mutants had no O antigen-containing capsule, but had 50% more lipopolysaccharide than did the parent, and this lipopolysaccharide had 30% fewer lipid A core molecules devoid of O antigen. Partially resistant mutants still had O antigen-containing capsule, but contained 40% more lipopolysaccharide than did the parent; the extent of coverage of lipid A core with O antigen remained unchanged. No correlations were found between outer membrane protein composition and the degree of serum resistance. Since the terminal membrane attack complex (C5b-9) must stably insert into a hydrophobic membrane site to effect killing, we conclude that both increased lipid A core and increased coverage of lipid A core with O antigen preclude access of C5b-9 to lethal sites on the cell surface.

Antigens, Bacterial↗
PubMed1976

Distribution of plasma cells and other cells containing immunoglobulin in the respiratory tract of normal man and class of immunoglobulin contained therein.

The anatomical distribution of plasma cells and other cells containing immunoglobulin in the respiratory tract, and the relative proportions of the immunoglobulin classes have been estimated on necropsy tissues from nine adult human subjects without respiratory disease, five non-smokers and four smokers, none of whom had cough or sputum. Cell counts on multiple sections stained by immunofluorescent methods for the presence of immunoglobulin were carried out on the upper trachea, main bronchus, and lower lobe bronchus. Cells containing immunoglobulin were found mostly in the submucous glands but were also present in the lamina propria of the tracheal and bronchial epithelium. These cells were present in the greatest concentration in the main bronchus and were always present in the lobar bronchus and, in most subjects, in the upper trachea. The cells were not always present round small bronchi and bronchioles and were virtually absent from alveolar walls. Cells containing IgA were much more numerous than those containing other immunoglobulin classes in all subjects except one, in whom IgG and IgE cells were equally numerous. Two subjects appeared to be significantly different from the rest. One non-smoking subject had a marked deficiency of IgA cells at all sampling sites, and one smoker had a marked excess of IgA cells. In spite of these two subjects there was no significant difference between smokers and non-smokers except in the lobar bronchus where the smokers had significantly more IgA cells than the non-smokers.

Adult↗
PubMed2001

Supervision: a container-contained approach.

The authors illustrate an approach to the supervisory process as a learning experience for both supervisee and supervisor built on the containment of unconscious anxieties. It is argued that a core function of psychoanalytic supervision is to help contain the emotional turbulence and the unconscious anxieties arising and evolving in the two interacting domains of the analytic and the supervisory sessions. From this perspective, the analyst-patient interaction and that of the supervisee and supervisor can be understood as twin, tiered transformational arenas, the supervisory one being at the service of holding and grasping the roles the supervisee/analyst goes through as part of the analytic process. On the basis of detailed clinical material from a disturbed 7-year-old girl, the authors explore the interrelated issues and difficulties in containing anxieties and turbulence in both the analytic and the supervisory situation. When emotional containment is adequately handled, the supervision helps the understanding and development of the supervisee's use of his/her own personality as a treatment instrument, as advocated by Fleming and Benedek decades ago. The supervisory session thus furthers the resolution of clinical issues through symbol-formation, clinical sessions and supervision being twin domains for recording and understanding emotional evolution.

Child↗
PubMed1994

Effects of tetracycline-containing gel and a mixture of tetracycline and citric acid-containing gel on non-surgical periodontal therapy.

The purpose of this study was to assess the clinical and microbiological effects of a newly developed root conditioning gel system containing tetracycline or a mixture of tetracycline and citric acid on non-surgical periodontal therapy. Sixty-four (64) single-rooted teeth with a probing depth of 4 to 6 mm were randomly subjected to one of the following four treatments; 1) root planing alone (RP group); 2) tetracycline-containing gel alone (TCG group); 3) root planing plus tetracycline-containing gel (RP + TCG group); or 4) root planing plus a mixture of tetracycline and citric acid-containing gel (RP + TC-CAG group). Probing depth, attachment level, and tooth mobility were measured and the presence of dental plaque and gingival inflammation was recorded at baseline and after 2, 4, 8, and 12 weeks. Subgingival plaque samples from each site were collected at the same visits and examined with phase contrast microscopy for proportions of motile rods and spirochetes. Plaque index, gingival sulcus bleeding index (SBI), probing depth, and attachment level decreased significantly in all groups compared to the baseline values (P < 0.05). A significant decrease in probing pocket depth was noted after 12 weeks in RP + TC-CAG group compared to the other groups (P < 0.05). Significantly more gain in attachment was detected in the RP + TC-CAG group compared to the TCG group (P < 0.05). Tooth mobility scores also decreased later in the study. A significant decrease in the proportion of motile rods was found primarily in the RP + TC-CAG group.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMedDate not supplied

Use of cellulose ether containing excipients with microcrystalline cellulose for the production of pellets containing metformin hydrochloride by the process of extrusion-spheronization.

The project is concerned mainly with the evaluation of two cellulose ether containing excipients, Aquacoat WG and Avicel 955 MCC for the improved extrusion-spheronization of metformin hydrochoride. Factorially designed experiments subject to statistical analyses were employed and products obtained were evaluated by sieve, packing density and image analysis, scanning electron microscopy and dissolution testing at pH 6.8. Aquacoat WG did not improve the ease of spheronization of drug mixes containing microcrystalline cellulose wetted with the optimum level of water. However, Avicel 955 MCC, which is a new experimental excipient containing 95% microcrystalline cellulose and 5% methylcellulose, did aid ease of spheronization facilitating acceptable yield of good spheres with high drug loadings (70%). Avicel 955 MCC containing drug mixes were more tolerant to minor alterations in level of hydration and yielded spheres which showed a small retardation of drug release despite the very high solubility of metformin hydrochloride.

Cellulose↗
PubMedDate not supplied

Aeration time following ethylene oxide sterilization for reusable rigid sterilization containers: concentration of gaseous ethylene oxide in containers.

Because ethylene oxide (EO) gas is toxic to humans, restrictions have been imposed on its use for sterilization, specifying allowable levels of residual EO remaining in sterilized apparatus and materials. However, the aeration time that optimizes the removal of the remaining EO when a rigid sterilizing container is used for a vessel had not been identified. Therefore, polyvinyl chloride, which easily adsorbs EO, was placed in rigid sterilizing containers, and aeration was carried out after 1, 8, 12, 17, and 24 hours. After standard EO sterilization, the EO concentrations remaining in the air in the rigid containers were measured. The results indicate that a period of 17 hours of aeration is appropriate when a rigid sterilizing container is used.

Air↗